2-ethoxyethyl trifluoromethanesulfonate

Conditions: temp:60.01; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: In a reaction flask, sirolimus (10 g, 10.9 mmol), ethyl di-isopropyl amine (78 g, 603.5 mmol), methylene chloride (35 ml) and 2-ethoxyethyl triflate (20 g, 88 mmol), each having a quantity about 10 times of that used in Example 3B, were added into the flask, which is filled with nitrogen gas. The reaction mixture is heated to 60° C. and is agitated for one hour and twenty minutes. Then, it is cooled, added with ethyl acetate (500 ml) and aqueous solution of hydrochloric acid (1N, 500 ml) and is further agitated. After agitation, it is settled for separating the layers. The organic layer is collected and respectively washed with pure water (500 ml), saturated saline (400 ml). The washed organic layer is dried and concentrated. The residue is then separated and purified by silica gel column chromatography to obtain white product of Biolimus A9 (2.9 g, 2.9 mmol), having a yield of 26.8% only. were added into the flask, which is filled with nitrogen gas Then, it is cooled for separating the layers The organic layer is collected respectively washed with pure water (500 ml), saturated saline (400 ml) The washed organic layer is dried concentrated The residue is then separated purified by silica gel column chromatography

Conditions: temp:60.01; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: In a reaction flask, sirolimus (10 g, 10.9 mmol), ethyl di-isopropyl amine (78 g, 603.5 mmol), methylene chloride (35 ml) and 2-ethoxyethyl triflate (20 g, 88 mmol), each having a quantity about 10 times of that used in Example 3B, were added into the flask, which is filled with nitrogen gas. The reaction mixture is heated to 60° C. and is agitated for one hour and twenty minutes. Then, it is cooled, added with ethyl acetate (500 ml) and aqueous solution of hydrochloric acid (1N, 500 ml) and is further agitated. After agitation, it is settled for separating the layers. The organic layer is collected and respectively washed with pure water (500 ml), saturated saline (400 ml). The washed organic layer is dried and concentrated. The residue is then separated and purified by silica gel column chromatography to obtain white product of Biolimus A9 (2.9 g, 2.9 mmol), having a yield of 26.8% only. were added into the flask, which is filled with nitrogen gas Then, it is cooled for separating the layers The organic layer is collected respectively washed with pure water (500 ml), saturated saline (400 ml) The washed organic layer is dried concentrated The residue is then separated purified by silica gel column chromatography

Conditions: temp:60.01; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: In a reaction flask, sirolimus (1 g, 1.1 mmol), ethyl di-isopropyl amine (7.8 g, 60.3 mmol), methylene chloride (3.5 ml) and 2-ethoxyethyl triflate (2.0 g, 8.8 mmol) as previously made in Example 3A were added into the flask, which is filled with nitrogen gas. The reaction mixture is heated to 60° C. and is agitated for one hour and twenty minutes. Then, it is cooled, added with ethyl acetate (100 ml) and aqueous solution of hydrochloric acid (1N, 100 ml) and is further agitated. After agitation, it is settled for separating the layers. The organic layer is collected and respectively washed with pure water (100 ml), saturated saline (80 ml). The washed organic layer is dried and concentrated. The residue is then separated and purified by silica gel column chromatography to obtain white product of Biolimus A9 (0.48 g, 0.49 mmol), with a yield of 44.5%. were added into the flask, which is filled with nitrogen gas Then, it is cooled for separating the layers The organic layer is collected respectively washed with pure water (100 ml), saturated saline (80 ml) The washed organic layer is dried concentrated The residue is then separated purified by silica gel column chromatography

Conditions: temp:60.01; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: In a reaction flask, sirolimus (1 g, 1.1 mmol), ethyl di-isopropyl amine (7.8 g, 60.3 mmol), methylene chloride (3.5 ml) and 2-ethoxyethyl triflate (2.0 g, 8.8 mmol) as previously made in Example 3A were added into the flask, which is filled with nitrogen gas. The reaction mixture is heated to 60° C. and is agitated for one hour and twenty minutes. Then, it is cooled, added with ethyl acetate (100 ml) and aqueous solution of hydrochloric acid (1N, 100 ml) and is further agitated. After agitation, it is settled for separating the layers. The organic layer is collected and respectively washed with pure water (100 ml), saturated saline (80 ml). The washed organic layer is dried and concentrated. The residue is then separated and purified by silica gel column chromatography to obtain white product of Biolimus A9 (0.48 g, 0.49 mmol), with a yield of 44.5%. were added into the flask, which is filled with nitrogen gas Then, it is cooled for separating the layers The organic layer is collected respectively washed with pure water (100 ml), saturated saline (80 ml) The washed organic layer is dried concentrated The residue is then separated purified by silica gel column chromatography

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: In a round bottom flask containing a stirring bar was placed 9.0 g (100 mmol) of ethoxyethanol. The atmosphere in the flask was replaced with nitrogen by using a nitrogen bubbler. The flask was given 160 mL of methylene chloride and 23.3 mL (120 mmol) of 2,6-lutidine. The flask cooled with ice was given dropwise 20.2 mL (120 mmol) of trifluoromethanesulfonic acid anhydride over 20 minutes. After stirring for 1 hour, the reaction liquid was mixed with 20 mL of saturated solution of ammonium chloride. The resulting mixture was washed sequentially with 1N hydrochloric acid (100 mL), deionized water (100 mL), saturated solution of sodium hydrogen carbonate (100 mL), and saturated aqueous solution of sodium chloride (100 mL). The organic layer was separated and dried with anhydrous sodium sulfate. With the sodium sulfate filtered off, the solution was concentrated under reduced pressure. The residue underwent silica gel chromatography. Thus there was obtained 15.03 g (67.6% yields) of 2-ethoxyethyl triflate from the fraction in eluate of 20% ethyl acetate-hexane. In a round bottom flask containing a stirring bar The flask cooled with ice over 20 minutes the reaction liquid The resulting mixture was washed sequentially with 1N hydrochloric acid (100 mL), deionized water (100 mL), saturated solution of sodium hydrogen carbonate (100 mL), and saturated aqueous solution of sodium chloride (100 mL) The organic layer was separated dried with anhydrous sodium sulfate With the sodium sulfate filtered off the solution was concentrated under reduced pressure

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: In a round bottom flask containing a stirring bar was placed 9.0 g (100 mmol) of ethoxyethanol. The atmosphere in the flask was replaced with nitrogen by using a nitrogen bubbler. The flask was given 160 mL of methylene chloride and 23.3 mL (120 mmol) of 2,6-lutidine. The flask cooled with ice was given dropwise 20.2 mL (120 mmol) of trifluoromethanesulfonic acid anhydride over 20 minutes. After stirring for 1 hour, the reaction liquid was mixed with 20 mL of saturated solution of ammonium chloride. The resulting mixture was washed sequentially with 1N hydrochloric acid (100 mL), deionized water (100 mL), saturated solution of sodium hydrogen carbonate (100 mL), and saturated aqueous solution of sodium chloride (100 mL). The organic layer was separated and dried with anhydrous sodium sulfate. With the sodium sulfate filtered off, the solution was concentrated under reduced pressure. The residue underwent silica gel chromatography. Thus there was obtained 15.03 g (67.6% yields) of 2-ethoxyethyl triflate from the fraction in eluate of 20% ethyl acetate-hexane. In a round bottom flask containing a stirring bar The flask cooled with ice over 20 minutes the reaction liquid The resulting mixture was washed sequentially with 1N hydrochloric acid (100 mL), deionized water (100 mL), saturated solution of sodium hydrogen carbonate (100 mL), and saturated aqueous solution of sodium chloride (100 mL) The organic layer was separated dried with anhydrous sodium sulfate With the sodium sulfate filtered off the solution was concentrated under reduced pressure

Conditions: temp:60.01; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: In a reaction flask, sirolimus (10 g, 10.9 mmol), ethyl di-isopropyl amine (78 g, 603.5 mmol), methylene chloride (35 ml) and 2-ethoxyethyl triflate (20 g, 88 mmol), each having a quantity about 10 times of that used in Example 3B, were added into the flask, which is filled with nitrogen gas. The reaction mixture is heated to 60° C. and is agitated for one hour and twenty minutes. Then, it is cooled, added with ethyl acetate (500 ml) and aqueous solution of hydrochloric acid (1N, 500 ml) and is further agitated. After agitation, it is settled for separating the layers. The organic layer is collected and respectively washed with pure water (500 ml), saturated saline (400 ml). The washed organic layer is dried and concentrated. The residue is then separated and purified by silica gel column chromatography to obtain white product of Biolimus A9 (2.9 g, 2.9 mmol), having a yield of 26.8% only. were added into the flask, which is filled with nitrogen gas Then, it is cooled for separating the layers The organic layer is collected respectively washed with pure water (500 ml), saturated saline (400 ml) The washed organic layer is dried concentrated The residue is then separated purified by silica gel column chromatography

Conditions: temp:60.01; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: In a reaction flask, sirolimus (1 g, 1.1 mmol), ethyl di-isopropyl amine (7.8 g, 60.3 mmol), methylene chloride (3.5 ml) and 2-ethoxyethyl triflate (2.0 g, 8.8 mmol) as previously made in Example 3A were added into the flask, which is filled with nitrogen gas. The reaction mixture is heated to 60° C. and is agitated for one hour and twenty minutes. Then, it is cooled, added with ethyl acetate (100 ml) and aqueous solution of hydrochloric acid (1N, 100 ml) and is further agitated. After agitation, it is settled for separating the layers. The organic layer is collected and respectively washed with pure water (100 ml), saturated saline (80 ml). The washed organic layer is dried and concentrated. The residue is then separated and purified by silica gel column chromatography to obtain white product of Biolimus A9 (0.48 g, 0.49 mmol), with a yield of 44.5%. were added into the flask, which is filled with nitrogen gas Then, it is cooled for separating the layers The organic layer is collected respectively washed with pure water (100 ml), saturated saline (80 ml) The washed organic layer is dried concentrated The residue is then separated purified by silica gel column chromatography

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: Coupling of 2-ethoxyethyl triflate (44.3 equivalents) and rapamycin (1.0 equivalent) in toluene with 2,6-lutidine (49.1 equivalents) was conducted at 55° C. (external water bath temperature) for 90 minutes. In this process (on a 2.0 g scale of, rapamycin), the 2-ethoxyethyl triflate was added to a pre-heated reaction mixture (55° C.) in one portion. After heating for 90 minutes, the external heating was turned off followed by addition of the second portion of 2,6-lutidine (54.9 equivalents). After the addition of the second lutidine portion, the reaction mixture was stirred for an additional 90 minutes at ambient temperature followed by workup to isolate crude BA9. The work-up included: 1. dilution of the reaction mixture with ethyl acetate; 2. quench of the reaction mixture with cold 1 N HCl; and 3. wash of the organic layer with three portions of 20% sodium chloride solution in water. The pH of the water layer after the last sodium chloride wash was ˜6. The crude material was then purified by an isocratic silica gel column chromatography using only one solvent mixture (n-hexane/ethyl acetate in ratio of 4:6 (vol/vol)). was conducted at 55° C. for 90 minutes After heating for 90 minutes the external heating to isolate crude BA9 quench of the reaction mixture with cold 1 N HCl wash of the organic layer with three portions of 20% sodium chloride solution in water The pH of the water layer after the last sodium chloride wash The crude material was then purified by an isocratic silica gel column chromatography

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: Coupling of 2-ethoxyethyl triflate (44.3 equivalents) and rapamycin (1.0 equivalent) in toluene with 2,6-lutidine (49.1 equivalents) was conducted at 55° C. (external water bath temperature) for 90 minutes. In this process (on a 2.0 g scale of, rapamycin), the 2-ethoxyethyl triflate was added to a pre-heated reaction mixture (55° C.) in one portion. After heating for 90 minutes, the external heating was turned off followed by addition of the second portion of 2,6-lutidine (54.9 equivalents). After the addition of the second lutidine portion, the reaction mixture was stirred for an additional 90 minutes at ambient temperature followed by workup to isolate crude BA9. The work-up included: 1. dilution of the reaction mixture with ethyl acetate; 2. quench of the reaction mixture with cold 1 N HCl; and 3. wash of the organic layer with three portions of 20% sodium chloride solution in water. The pH of the water layer after the last sodium chloride wash was ˜6. The crude material was then purified by an isocratic silica gel column chromatography using only one solvent mixture (n-hexane/ethyl acetate in ratio of 4:6 (vol/vol)). was conducted at 55° C. for 90 minutes After heating for 90 minutes the external heating to isolate crude BA9 quench of the reaction mixture with cold 1 N HCl wash of the organic layer with three portions of 20% sodium chloride solution in water The pH of the water layer after the last sodium chloride wash The crude material was then purified by an isocratic silica gel column chromatography

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: To a stirred solution of 1 g rapamycin, and 7.66 g 2,6-lutidine in 14.65 mL toluene held at 60° C. was added 5.81 g 2-ethoxyethanol triflate. Stirring was continued for 90 minutes after which 50 mL ethyl acetate was added to the reaction and the solution was washed with 50 mL 1M HCl. The organic material was washed with D.I. water until pH of wash solution was neutral and the organic solution was dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by flash chromatography on silica gel, 200–400 mesh (40:60 hexane-ethyl acetate (v/v) to give 210 mg 42-O-(2-ethoxyethyl) rapamycin. TLC Rf=0.41 using 40:60 hexane-ethyl acetate (v/v). MS (ESI) m/z 1008.5 C55H87NNaO14. A mass spectrum of the title compound is shown in FIG. 1. held at 60° C. the solution was washed with 50 mL 1M HCl The organic material was washed with D.I the organic solution was dried over anhydrous sodium sulfate filtered concentrated The residue was purified by flash chromatography on silica gel, 200–400 mesh (40:60 hexane-ethyl acetate (v/v)

Conditions: temp:-5.01; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: 25 g of ethoxyethanol was placed in a round bottom flask provided with a stirrer piece therein, which was connected to a nitrogen bubbler to replace the air in the flask by dry nitrogen. 141 ml of methylene chloride was added and 33 ml of 2,6-lutidine was further added, followed by cooling down to −10 to 0° C., dropping 57 ml of trifluoromethanesulfonic acid anhydride in one hour and continuing agitation under the cooling conditions for one hour. 100 ml of distilled water was added to the reaction solution, and an organic phase was separated, washed with 100 ml of distilled water and dried with anhydrous sodium sulfate, followed by removing the sodium sulfate by filtration and distilling off the solvent under reduced pressure. The resulting residue was distilled under a reduced pressure of 150 Pa at 30° C. to obtain 57 g of a 2-ethoxyethyl triflate fraction. provided with a stirrer piece in one hour for one hour an organic phase was separated washed with 100 ml of distilled water dried with anhydrous sodium sulfate by removing the sodium sulfate by filtration distilling off the solvent under reduced pressure The resulting residue was distilled under a reduced pressure of 150 Pa at 30° C.