[ CAS No. 159351-69-6 ] {[proInfo.proName]}

Name: 159351-69-6|(3S,6R,7E,9R,10R,12R,14S,15E,17E,19E,21S,23S,26R,27R,34aS)-9,27-dihydroxy-3-((R)-1-((1S,3R,4R)-4-(2-hydroxyethoxy)-3-methoxycyclohexyl)propan-2-yl)-10,21-dimethoxy-6,8,12,14,20,26-hexamethyl-9,10,12,13,14,21,22,23,24,25,26,27,32,33,34,34a-hexadecahydro-3H-23,27-epoxypyrido[2,1-c][1]oxa[4]azacyclohentriacontine-1,5,11,28,29(4H,6H,31H)-pentaone|98%
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Quality Control of [ 159351-69-6 ]

COA NMR CNMR HPLC LC-MS

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SDS Specification

Alternatived Products of [ 159351-69-6 ]

Product Details of [ 159351-69-6 ]

CAS No. :	159351-69-6	MDL No. :	MFCD00929329
Formula :	C₅₃H₈₃NO₁₄	Boiling Point :	-
Linear Structure Formula :	-	InChI Key :	-
M.W :	958.22	Pubchem ID :	-
Synonyms :	RAD001;SDZ-RAD;Xience V.;Certican;Zortress;Brand name Afinitor;RAD 001.SDZ-RAD	Chemical Name :	(3S,6R,7E,9R,10R,12R,14S,15E,17E,19E,21S,23S,26R,27R,34aS)-9,27-dihydroxy-3-((R)-1-((1S,3R,4R)-4-(2-hydroxyethoxy)-3-methoxycyclohexyl)propan-2-yl)-10,21-dimethoxy-6,8,12,14,20,26-hexamethyl-9,10,12,13,14,21,22,23,24,25,26,27,32,33,34,34a-hexadecahydro-3H-23,27-epoxypyrido[2,1-c][1]oxa[4]azacyclohentriacontine-1,5,11,28,29(4H,6H,31H)-pentaone

Safety of [ 159351-69-6 ]

Signal Word:	Warning	Class:	N/A
Precautionary Statements:	P280-P305+P351+P338	UN#:	N/A
Hazard Statements:	H302	Packing Group:	N/A
GHS Pictogram:

Application In Synthesis of [ 159351-69-6 ]

* All experimental methods are cited from the reference, please refer to the original source for details. We do not guarantee the accuracy of the content in the reference.

Upstream synthesis route of [ 159351-69-6 ]
Downstream synthetic route of [ 159351-69-6 ]

[ 159351-69-6 ] Synthesis Path-Upstream 1~7

1
[ 1314705-20-8 ]
[ 159351-69-6 ]

Yield	Reaction Conditions	Operation in experiment
15%	With pyridine; hydrogen fluoride In tetrahydrofuran at 0 - 20℃; for 4 h;	To a solution of intermediate 17 (280 mg. 0.23 mmol) in THF (10 mL) was added 2 mL HF in pyridine at 0 deg and stirred at rt for 4 hours. Then 20 ml water ws added and extracted with EtOAc (20 mL x 3). The combined organic layer was washed by 0.5N HCI saturated NaHCO3 and brine, dried over anhydrous Na2S04. After concentration the residue was purified with silica gel chromatography (30percent to 100 percent of EtOAc in petroleum ether as eluent) to give white solid which was further purified by prep-HPLC to give compound A15 (34 mg. 15percent) as a white solid.
15%	With pyridine hydrogenfluoride In tetrahydrofuran at 0 - 20℃; for 4 h;	To a solution of intermediate 17 (280 mg, 0.23 mmol) in THF (10 mL) was added 2 mL HF in pyridine at 0 deg and stirred at rt for 4 hours. Then, 20 mL water was added and extracted with EtOAc (20 mL×3). The combined organic layer was washed by 0.5N HCl, saturated NaHCO₃and brine, dried over anhydrous Na₂SO₄. After concentration, the residue was purified with silica gel chromatography (30percent to 100percent of EtOAc in petroleum ether as eluent) to give white solid which was further purified by prep-HPLC to give compound A15 (34 mg, 15percent) as a white solid. ¹H NMR (300 MHz, CDCl₃) δ6.40-6.00 (m, 5H), 5.53-5.25 (m, 4H), 4.83 (s, 1H), 4.13 (m, 1H): LCMS (m/z) ES− 957 (M−1)⁻.
6.81 g	With pyridine hydrogenfluoride In tetrahydrofuran at 0 - 45℃; for 3.5 h;	To a solution of 2-((tert-butyldiphenylsilyl)oxy)ethanol (13.1 g, 43.8 mmol) in toluene (51 g) was added N,N-Diisopropylpentan-3-amine (8.7 g, 50.3 mmol) The clear solution was then cooled to 0°C and trifluoromethanesulfonic acid anhydride (12.3 g, 43.8 mmol) was added dropwise such that the temperature was maintained between (-2°C - 2°C). Following the addition a further portion of toluene (5 g) was used for washing. After 1 .5 h N,N-Diisopropylpentan-3-amine (8.7 g, 50.3 mmol) was added followed by toluene (3 g) and Rapamycin (10.0 g, 10.9 mmol ) washing with toluene (18 g). The reaction was then heated to 40°C and allowed to stir at this temperature for 22.5 h at which point less than 5 Areapercent Rapamycin was remaining according to HPLC analysis. The reaction was cooled to ambient temperature and pyridine (1 .0 mL) was then added to quench the reaction which was stirred for a further 30 mins. The reaction was filtered and diluted with isopropyl acetate. The organic solution was washed with 1 M citric acid solution, 10percent sodium bicarbonate solution followed by water, dried (MgS0₄) and concentrated in vacuo. To the crude residue (35.6 g) was added THF (240 mL) and this solution was then added dropwise at 0 °C to a HF*pyridine solution (1 :1 , 38.1 g). The reaction was heated to 45 °C for 3.5 h then allowed to cool to ambient temperature and diluted with isopropylacetate (300 g). The reaction was then added slowly to an 8percent aqueous solution of sodium bicarbonate and further washed with isopropylacetate (250 g). The organic phase was then separated and washed with saturated aqueous sodium chloride solution, dried (MgS0₄) and concentrated in vacuo. The residue was diluted with isopropylacetate, BHT (0.2percent m/m) was added, and the yield of everolimus determined by HPLC analysis against an external standard (6.81 g, 65percent).

Reference： [1] Patent: WO2014/82286, 2014, A1, . Location in patent: Paragraph 50
[2] Patent: US2015/51242, 2015, A1, . Location in patent: Paragraph 0086
[3] Patent: WO2014/203185, 2014, A1, . Location in patent: Page/Page column 22; 23; 24; 26

2
[ 159351-68-5 ]
[ 159351-69-6 ]

Yield	Reaction Conditions	Operation in experiment
2.96 g	With pyridine hydrogenfluoride In tetrahydrofuran at 0 - 45℃; for 1.5 h;	To a solution of 2-((tert-butyldimethylsilyl)oxy)ethanol (8.04 g, 43.8 mmol ) in toluene (55 g) was added N,N-diisopropylethylamine (5.94 g, 45.9 mmol ) The clear solution was then cooled to 0°C and Trifluoromethanesulfonic acid anhydride (1 1 .97 g, 42.4 mmol) was added dropwise such that the temperature was maintained between (-2°C - 2°C). Following the addition a further portion of toluene (5 g) was used for washing. After 30 minutes N,N-Diisopropylpentan-3-amine (7.871 g, 45.9 mmol) was added followed by toluene (3 g) and Rapamycin (10.0 g, 10.9 mmol) washing with toluene (18.4 g). The reaction was then heated to 40 °C and allowed to stir at this temperature for 42 h at which point less than 5 Areapercent Rapamycin was remaining according to HPLC analysis. The reaction was cooled to ambient temperature and pyridine (2.6 g) was then added to quench the reaction which was stirred for a further 30 mins. The reaction was filtered and diluted with isopropyl acetate. The organic solution was washed with 1 M citric acid solution, 10percent sodium bicarbonate solution followed by water, dried (MgS0₄) and concentrated in vacuo. The residue was divided into two portions. To half of this crude residue (9.66 g) THF (100 ml_) was added and this solution was then added dropwise at 0 °C to a HF*pyridine solution (1 :1 , 17.7 g). A further portion of THF (20 ml_) was used for washing. The reaction was heated to 45 °C for 1 .5 h, then allowed to cool to ambient temperature and diluted with isopropylacetate (150 g). The reaction was then added slowly to an 8percent aqueous solution of sodium bicarbonate and further washed with isopropylacetate (250 g). The organic phase was then separated and washed with saturated aqueous sodium chloride solution, dried (MgS0₄) and concentrated in vacuo. The residue was diluted with isopropylacetate, butylhydroxytoluol (BHT; 0.2percent m/m) was added, and the yield of everolimus determined by HPLC analysis against an external standard (2.96 g, 57percent).
68 %Chromat.	With hydrogenchloride; phosphoric acid; water In n-heptane; acetonitrile at 20℃; for 1 h;	Example 2: Synthesis of 40-O-(2-hydroxy)ethyl-rapamycin (i.e. everolimus) The crude 40-O-[2-(f-butyldimethylsilyl)oxy]ethyl-rapamycin solution of Example 1 was evaporated to dryness under reduced pressure at 25-30° C. Subsequently, 66 ml of heptane were added to the residue, and the resulting mixture was evaporated to a volume of 40 ml. Then, 48 ml of a (80:20) (v/v) mixture of acetonitrile/water (pH = 1.7, adjusted with 75percent ortho-phosphoric acid) were added, and the pH of the resulting mixture was adjusted to 1 .8 with 1 N HCI solution. The resulted mixture was stirred for 1 hour at room temperature, and the lower layer containing everolimus was separated. The overall yield of everolimus as determined by HPLC using an external standard was 68percent (71 1 mg).

Reference： [1] Patent: WO2014/203185, 2014, A1, . Location in patent: Page/Page column 21; 22
[2] Patent: WO2016/20664, 2016, A1, . Location in patent: Page/Page column 22
[3] Patent: CN103848849, 2016, B, . Location in patent: Paragraph 0027; 0030; 0031
[4] Patent: WO2016/207205, 2016, A1, . Location in patent: Page/Page column 13

3
[ 75-21-8 ]
[ 53123-88-9 ]
[ 159351-69-6 ]

Yield	Reaction Conditions	Operation in experiment
6.9 g	With trifluorormethanesulfonic acid In toluene at 70℃; for 8 h; Autoclave	9.14 g rapamycin (0.01mol), 11g ethylene oxide (0.25 mol), 0.1 g trifluoromethanesulfonic acid, 30mL toluene was added to the autoclave after mixing, Warmed to 70 ° C, Maintaining 1.0 MPa pressure, the reaction was stopped after 8 hours, cooled to room temperature,The solvent was recovered under reduced pressure and the residue was purified by silica gel column chromatography (200-300 mesh silica gel,Eluent: ethyl acetate: petroleum ether = 20: 1),7.7 g 97.95percent of everolimus was obtained, which was purified by HP-20 resin column chromatography (eluent: acetonitrile: water = 65:35) 6.9 g of everolimus was obtained as a white solid, HPLC purity:99.6percent, Isomer content:0.12percent. Molar yield:72.0percent

Reference： [1] Patent: CN104876944, 2017, B, . Location in patent: Paragraph 0035-0050

4
[ 1392400-31-5 ]
[ 159351-69-6 ]

Reference： [1] Patent: WO2012/103959, 2012, A1, . Location in patent: Page/Page column 18
[2] Patent: CN106478663, 2017, A, . Location in patent: Paragraph 0023

5
[ 107-21-1 ]
[ 53123-88-9 ]
[ 159351-69-6 ]

Yield	Reaction Conditions	Operation in experiment
5.7 kg	Stage #1: With trifluoroacetic anhydride In tetrahydrofuran at 10℃; for 1.5 h; Inert atmosphere; Large scale Stage #2: With boron trifluoride diethyl etherate In tetrahydrofuran at 10℃; for 2 h; Large scale	620g of ethylene glycol and 6L of tetrahydrofuran were added to the reaction flask,Mix well to mix.The reaction temperature was controlled at 10 .Under nitrogen protection,1.41L trifluoroacetic anhydride was slowly added dropwise,Dropping is completed,Reaction for 1.5 hours,The reaction solution.9.14 kg of rapamycin was dissolved in 54 L of tetrahydrofuran,Added to the reaction solution,The reaction temperature was controlled at 10 .Slowly add 13ml boron trifluoride diethyl ether solution. Bi completed,The reaction was stirred for 2 hours. After the reaction is completed,60L saturated aqueous sodium bicarbonate solution was added,Stir wellThen suction filtered,To the filtrate was added 30 L of ethyl acetate,Liquid separation,The organic phase is washed with pure water until nearly neutral.The organic phase was dried over 500 g of anhydrous sodium sulfate for 2 hours, filtered,Concentrated under reduced pressure to a solventless outflow,A thick liquid. Column chromatography,The eluent is petroleum ether:Ethyl acetate = 1: 6. The collected effluent was concentrated under reduced pressure to give 6.3 kg of yellow foamy solid,Yield 66percent.A mixture of 26.8 L of methanol and ethyl acetate (v / v = 1/3) was added to the above yellow foamy solid,Stirring to dissolve,The temperature was controlled at 25 for 30 minutes,13.4 L cyclohexane was added dropwise,Bi completed,The temperature was controlled at 12 for 2 hours,Cool the feed liquid to about 0 slowly stirring 3h,Suction filtration,Drying at room temperature under vacuum gave 5.7 kg of a white solid,HPLC and mass spectrometry determined that the white solid was everolimus,Purity 98.1percent.

Reference： [1] Patent: CN106146535, 2016, A, . Location in patent: Paragraph 0037; 0038; 0039; 0040; 0041; 0042; 0043-0056

6
[ 53123-88-9 ]
[ 159351-69-6 ]

Reference： [1] Patent: WO2014/203185, 2014, A1,
[2] Patent: WO2016/20664, 2016, A1,
[3] Patent: CN105254646, 2016, A,
[4] Patent: CN105254646, 2016, A,
[5] Patent: CN105254646, 2016, A,
[6] Patent: CN105254646, 2016, A,
[7] Patent: WO2016/207205, 2016, A1,
[8] Patent: CN106146536, 2016, A,

7
[ 331810-13-0 ]
[ 159351-69-6 ]

Reference： [1] Patent: CN106146536, 2016, A,

[ 159351-69-6 ] Synthesis Path-Downstream 1~55

1
[ 64-69-7 ]
[ 159351-69-6 ]
SDZ-RAD-iodoacetate ester [ No CAS ]

Yield	Reaction Conditions	Operation in experiment
73%	With dmap; dicyclohexyl-carbodiimide; In dichloromethane; at 0 - 20℃;Inert atmosphere; Darkness;	<strong>[159351-69-6]Everolimus</strong> (5.0g, 5.2mmol) and iodoacetic acid (0.77g, 4.16mmol) were dissolved in dichloromethane (50 mL). The obtained mixture was cooled to 0-5C, dicyclohexylcarbodiimide (1.07g, 5.2mmol) and 4-dimethylaminopyridine (0.63g, 1.58mmol) were added to the mixed solution successively. Then the reaction mixture was warmed to room temperature and stirred for about 3 hours under a nitrogen atmosphere in darkness. Then, the reacted mixture was cooled to 0-5C. Iodoacetic acid (0.48g, 2.6mmol) and dicyclohexylcarbodiimide (0.86g, 4.16mmol) were added to the reaction successively, and stirred for another 3h under a nitrogen atmosphere in darkness. After reaction finished, the suspension was filtered with a funnel and the filtrate was concentrated in vacuo. The residue was purified by silica gel column chromatography (n-hexane:acetone=9:1 to 3:1) to obtain compound 2 (4.26g, 3.79mmol) as a white powder. M.p. 68.5-70.3C; [alpha]20D [alpha]D20 -93.9 (c=1.0, MeOH); IR (KBr) numax cm-1: 3419, 3327, 2929, 2852, 1733, 1627; 1H NMR (400 MHz, CDCl3) delta 6.35 (dt, J=24.8, 14.8 Hz, 2H), 6.19-6.09 (m, 1H), 5.93 (dd, J=30.1, 10.5 Hz, 1H), 5.60-5.45 (m, 1H), 5.41 (d, J=9.9 Hz, 1H), 5.27 (t, J=7.2 Hz, 1H), 5.15 (dt, J=11.6, 5.5 Hz, 1H), 4.34-4.24 (m, 2H), 4.16 (ddd, J=21.4, 14.1, 6.6 Hz, 2H), 3.91-3.75 (m, 3H), 3.73 (d, J=7.1 Hz, 2H), 3.67 (dd, J=14.5, 6.8 Hz, 1H), 3.57 (d, J=13.5 Hz, 1H), 3.44 (d, J=10.3 Hz, 4H), 3.40-3.28 (m, 4H), 3.21-3.00 (m, 5H), 2.84 (dd, J=17.7, 7.0 Hz, 1H), 2.72 (dd, J=16.4, 5.5 Hz, 2H), 2.58 (dd, J=16.7, 6.4 Hz, 1H), 2.33 (d, J=12.9 Hz, 2H), 2.10-1.89 (m, 6H), 1.75 (s, 6H), 1.71-1.57 (m, 8H), 1.54-1.40 (m, 4H), 1.38-1.19 (m, 8H), 1.12 (dd, J=19.6, 6.8 Hz, 4H), 1.05 (d, J=6.3 Hz, 4H), 0.99 (d, J=6.4 Hz, 2H), 0.95 (d, J=6.5 Hz, 2H), 0.93-0.82 (m, 5H), 0.71 (dt, J=16.5, 8.3 Hz, 1H); 13C NMR (100 MHz, CDCl3) delta 215.4, 208.1, 192.6, 171.1, 169.8, 169.2, 168.7, 166.7, 165.7, 140.1, 136.0, 133.6, 130.1, 129.5, 129.2, 126.7, 126.4, 98.7, 83.2, 75.6, 67.6, 67.1, 65.7, 60.3, 59.3, 57.9, 55.9, 51.2, 49.2, 46.5, 44.2, 41.4, 40.6, 38.3, 36.3, 35.1, 33.8, 33.1, 31.7, 30.1, 27.2, 27.0, 25.5, 25.2, 24.8, 21.5, 21.0, 20.6, 16.2, 15.8, 14.1, 13.7, 13.1, 10.1, 5.5; HR-ESI-MS m/z: 1148.4787 [M+H]+ (calcd. for C55H84INO15 1148.4778).
	With dmap; dicyclohexyl-carbodiimide; In dichloromethane; at 20℃; for 2.66667h;	SDZ-RAD (0.50 g 5.2 x 104 mole), 4-Dimethylaminopyridine (3.0 mg) and 1,3-Dicyclohexylcarbodiimide (0.136 g, 6.6 x 10-4 mole) were dissolved in 20 ml anhydrous methylene chloride in a 150 ml round-bottom flask. Iodoacetic acid (0.116 g, 6.3 x 10-4 mole) was dissolved in 10 ml anhydrous methylene chloride. The iodoacetic acid solution was added into the reaction mixture over a period of 10 min. with stirring by a magnetic bar. Then the reaction mixture was stirred at room temperature for another 2.5 h. The solution was then filtered through a sintered glass filter. The filtrate was transferred to a separatory funnel, washed 50 ml of sodium bicarbonate solution (5.5 g/ 100 ml) and then washed with 2 x 50 ml of water. The methylene chloride layer was dried with 5g anhydrous sodium sulfate for 4 hours. Then sodium sulfate was filtered out and methylene chloride was removed by rotary evaporation. A total of 0.60g pale yellow solid was obtained. Isolation of pure SDZ-RAD-iodoacetate ester was performed by preparative HPLC on a Prep Nova-pak HR C18 (300 x 19 mm) column from Waters. SDZ-RAD-iodoacetate ester eluted at 18.4 min using a gradient ( 30% A, 70% B for 5 min. then to 100% B in 30 min.). A is 90% water, 10% acetonitrile; B is 10% water, 90% acetonitrile. The fraction was collected and extracted by 2 x 50 ml methylene chloride. The organic layer was combined and dried with anhydrous sodium sulfate for 4 h. The organic solvent was removed by rotary evaporation to dryness. A yellowish solid was obtained (0.102 g).1H NMR (CDCL3, 400 MHz) d 3.72 (s, 2H, I-CH2-CO2-), 4.28 (m, 2H, -CO2-CH2-). MS m/z 1134.6 (M+NH4)+; m/z 1184.6 (M+OAC)-. MS/MS 213.0 (ICH2CO2CH2CH2)+.

Reference： [1]Bioorganic and Medicinal Chemistry Letters,2017,vol. 27,p. 1175 - 1178
[2]Patent: EP1210350,2004,B1 .Location in patent: Page 7

Yield	Reaction Conditions	Operation in experiment
	Purification / work up;	The procedure below is carried-out at 28C to 32 C. A bed of sorption resin (AMBERLITE XAD 1180) in a column (45 cm diameter) using water: THF to charge the column (ca. 100 L wet sorption resin) is prepared. Water (86 L) is slowly added, with agitation, to a solution of everolimus (1227 g) in acetonitrile (10 L) in which sorption resin (AMBERLITE XAD 1180; 9 L) is suspended with stirring. The everolimus used contains impurities. When the addition of water is complete, the loading charge of sorption resin is collected by filtration. The collected loading charge is loaded (juxtaposed) as a layer on top of the bed of wet sorption resin. The column is first eluted with ca. 1800 L of a first eluent made-up of THF/water (33 vol-% THF). The column is then eluted with second eluent made-up of THF/water (40 vol-% THF). The elution rate is about 11 to 13 L/hr (6.9 to 8. 2 cm/hr). A main fraction, ca 460 L containing everolimus is collected. A pre-fraction, ca. 80 L, containing everolimus, is also collected. The main fraction (460 L) is combined with phosphoric acid, 85% (460 mL), and concentrated at reduced pressure to a volume of about 230 L. The concentrate is held at ambient temperature for one day. The crystals are washed with hexane and dried at 40 C.
	Purification / work up;	Two columns are prepared for chromatography according to example 1. Before the chromatography, 3000G active substance containing everolimus is adsorbed on sorption resin XAD 1180 according to the following procedure. The everolimus is dissolved in 15L acetone. Sorption resin (33 L) is added to the solution, and 90L water is added slowly to the solution/resin mixture with continuous stirring. The loading charge of sorption resin is headed (juxtaposed) as a layer on the top of sorption resin contained in the first column. The first column is eluted with tetrahydrofuran : water mixture (34vol-% THF). The elution rate is 15 L/hour. Fractions of 20 L each are collected. Volume of each fraction was 20L. After elution of the 35th fraction, the second column is connected (fluidly coupled) in series to the first column, and the elution is continued on columns in series. After the 95th fraction eluted, the first column is disconnected, and the elution is continued only on the second column. The purified, suitable fractions are combined. The major part of THF is removed from the combined fractions by evaporation under reduced pressure. The concentrate is extracted with ethylacetate and the phases separated. The separated ethylacetate phase is concentrated under reduced pressure (appr. L part everolimus and 1 part ethylacetate). Cyclohexane and water are added slowly to the concentrated ethyl acetate extract. The precipitated everolimus is recovered from the mixture at 0-30C. The crystals are filtered and dried.
		, wherein the rapamycin derivative is selected from the group consisting of rapamycin, 40-O-(2-hydroxy)ethyl-rapamycin, 40-O-(3-hydroxy)propyl-rapamycin, 40-O-[2-(2-hydroxy)ethoxy]ethyl-rapamycin, and 40-O-(2-acetaminoethyl)-rapamycin.

, wherein the rapamycin derivative is selected from the group consisting of 40-O-(2-hydroxy)ethyl-rapamycin, 40-O-(3-hydroxy)propyl-rapamycin, 40-O-[2-(2-hydroxy)ethoxy]ethyl-rapamycin, and 40-O-(2-acetaminoethyl)-rapamycin.

3
[ 159351-69-6 ]
42-O-(2-hydroxy)ethyl rapamycin [ No CAS ]

Yield	Reaction Conditions	Operation in experiment
	With triethylammonium acetate; In 1,4-dioxane; water; at 22 - 28℃; for 4h;	B. Preparation of Compound SDZ-RAD (40 mg; 0.042 mmol) was dissolved in 50 mL 40% 0.1 M TEAA buffer pH 8.5 and 60% dioxane. The solution was stirred at room temperature (approximately 22 C. to 28 C.) for 4 hours. The conversion was stopped by 2×50 mL CH2Cl2 extraction. The organic layer was reduced by rotary evaporation system to dryness. The isolation of compound was performed by BDS-Hypersil-C18 column (250×20 mm) using the mobile phase consisting of 60% dioxane, 40% water with 0.01M TEAA buffer pH 3.9. The flow rate was 12 mL/min. The fraction of isomer C (28.9 min) was collected and extracted with CH2Cl2 using a separatory funnel. The organic layer was combined and washed with 2×50 mL water and then the organic layer was dried with anhydrous Na2SO4. The organic solvent was reduced by rotary evaporation to about 1 mL. The product was transferred into a vial and precipitated by adding n-hexane. White powder was obtained by using N2 to blow away the solvent and the vial was put into speed-vac to dry overnight. The purity of the isomer C for each purification step was analyzed by HPLC. ESI mass spectrometry indicates molecular ion [M+NH4]+ m/e 975.8, which is same as the reference sample of SDZ-RAD. The NMR sample was prepared in acetone-d6. Table: Proton and Carbon resonance assignment of SDZ-RAD isomer-C (NB No. L20156-xxx) in Aceton-d6 at 25 C., 500 MHz (13C: 100 MHz) delta 13Cdelta 1Hdelta 13C CNo. major major minordelta 1H minor1H Correltn HMBC 1 140.24 5.44 139.96 5.83 2, 36 2 131.87 6.23 130.94 6.26 3, 1 3 133.61 6.27 134.52 6.37 4, 2 4 127.90* 6.49 128.13* 6.48 3, 5 5 128.14 6.16 130.38 6.25 4, 45 6 139.63 137.02 7 84.16 3.77 84.60 3.88 8 5, 6, 8, 45, 52 8 44.00 2.14; 1.34 41.88 1.86; 1.72 7, 9 6, 7 9 74.60 3.79 73.30 3.48 8, 10 15 10 33.62 2.05eq 36.01 1.78eq 9, 11 1.52ax 1.71ax 11 35.54 1.82eq 35.28 1.75eq 10, 12 1.24ax 1.23ax 12 44.08* 3.24 44.15* 3.17 11 10, 46 13 210.92 210.43 15 98.77 99.07 15 6.07 5.86 12, 13, 15, (OH) 16 16 168.84 168.52 18 44.15 4.60eq 39.87 4.16eq 19 3.02/168.52 2.93ax 3.02ax 19 26.04 1.57 25.54 1.74 18, 20 1.43 1.44 20 21.78* 1.66 21.69* 1.79 19, 21 1.51 1.45 21 27.09 2.28 28.47 2.28 20, 22 1.52 1.87 22 52.54 5.07 56.58 5.58 21 23 170.66 171.35 25 75.95 5.25 75.59 5.27 26, 37 23, 26, 28, 37, 38, 51 26 41.46 2.80 42.84 2.83 25 2.66 2.74 27 208.50 208.60 28 46.78 3.40 46.89 3.47 29, 47 47 29 126.08 5.31 127.55 5.52 28, 48 28, 47 30 137.83 138.27 31 77.63* 4.21 77.69* 4.22 29, 32, 48 31 (OH) 4.05 4.05 30, 31, 32 32 85.96 4.16 87.57 3.82 31, 32, 34 33 211.79 212.16 34 42.67 2.56 40.39 2.93 35, 49 35 40.70 1.57(proR) 41.99 1.55(proR) 34, 36 1.16(proS) 1.25(proS) 36 37.2 2.29 34.87 2.37 1, 35, 50 37 34.30 1.86 34.43 1.94 38, 51 38 40.10 1.36 39.16 1.26 37, 39 1.08 1.13 39 33.80* 1.36 33.96* 1.40 38, 40, 44 40 37.46 2.04 37.07 2.06 39, 41 39, 44 0.73 0.69 41 84.14* 3.12 84.27* 3.02 40, 42 42 83.80* 3.11 83.75 3.11 41, 43 43 31.24* 2.01 31.19* 2.01 42, 44 1.21 1.21 44 32.38 1.68 32.03 1.72 39, 43 0.93 0.91 45 10.74 1.72 10.99 1.67 5 46 17.17* 1.13 17.14* 1.14 12 11, 12, 13 47 15.96 0.95 17.02 0.09 28 48 14.56 1.83 13.42 1.75 29 49 14.32 0.93 14.94 1.02 34 50 22.04 1.03 21.43 1.06 36 51 15.63 0.87 14.56 0.93 37 37 52 56.51 3.12 56.25 3.18 53 58.04 3.27 58.35 3.26 54 57.69 3.27 58.35 3.26 55 72.37* 3.58* 72.40* 3.65* 56 62.58 3.58 62.58 3.58

Reference： [1]Patent: US2006/199834,2006,A1 .Location in patent: Page/Page column 4-6

4
[ 5309-70-6 ]
[ 159351-69-6 ]
[ 159351-81-2 ]

Yield	Reaction Conditions	Operation in experiment
96%	Novozym 435TM lipase; In tert-butyl methyl ether; at 35℃; for 10h;Molecular sieve;	A. Preparation of <strong>[159351-69-6]Everolimus</strong> 42-bromoacetate (VIII) A mixture of <strong>[159351-69-6]everolimus</strong> (250 mg, 0.26 mmol), vinyl bromoacetate (165 mg, 1 mmol), 5 A molecule sieves (20 mg), and Novozym 435 lipase (200 mg) in anhydrous t-butyl methyl ether (TBME) (3 mL) was heated under N2 at 35 C. for 10 hours. The enzyme was removed by filtration and washed with TBME. The mixture was concentrated and triturated with hexane. The <strong>[159351-69-6]everolimus</strong> 42-bromoacetate (VIII) was collected by filtration and dried in vacuo. Yield: 275 mg (96%). MS (ESI) m/e 1078 (M-)
	at 20℃; for 4h;Enzymatic reaction;	<strong>[159351-69-6]Everolimus</strong> (1.0 g, 1.04 mmol) and vinyl bromoacetate(0.257 g, 1.56 mmol) were dissolved in dichloromethane(10 mL). Then the immobilized lipase (0.5 g) was added tothe mixture, which was stirred for about 4 h at room temperature.After reaction finished, the immobilized lipase wasfiltered with a funnel and the filtrate was concentrated invacuo to obtain compound 2 (1.12 g, 1.04 mmol) as a whitefluffy solid. Mp 64.6-66.3 C; [alpha]20D -121.5 (c 1.0,MeOH); IR (KBr) numax 3443, 2931, 2870, 1722, 1643cm-1; 1H NMR (DMSO-d6, 400 MHz): delta = 6.48 (1H, s),6.37-6.44 (1H, m), 6.19-6.25 (1H, m), 6.13 (2H, d, J =10.8 Hz), 5.49 (1H, q, J = 24.1 Hz), 5.25-5.27 (1H, m),5.10 (1H, d, J = 10.2 Hz), 4.93-4.96 (2H, m), 4.27 (2H, brs), 4.15 (2H, s), 4.01 (2H, br s), 3.96 (1H, d, J = 4.12 Hz),3.70-3.72 (2H, m), 3.64 (1H, d, J = 12.2 Hz), 3.42-3.45(3H, m), 3.24-3.28 (1H, m), 3.15 (3H, s), 3.09-3.12 (1H,m), 3.05 (3H, s), 2.98 (1H, s), 2.75 (1H, d, J = 17.1 Hz),2.34-2.40 (2H, m), 1.98-2.22 (1H, m), 2.12 (1H, d, J =12.7 Hz), 2.00-2.02 (1H, m), 1.85-1.92 (1H, m), 1.82-1.85(2H, m), 1.74 (2H, s), 1.68-1.70 (2H, m), 1.60-1.63 (4H,m), 1.46-1.54 (4H, m), 1.37-1.43 (2H, m), 1.24-1.29 (6H,m), 1.14-1.18 (2H, m), 1.01-1.06 (3H, m), 0.94-0.99 (4H,m), 0.86-0.87 (5H, m), 0.81-0.84 (3H, m), 0.78 (2H, d, J =6.4 Hz), 0.74 (3H, d, J = 6.4 Hz); 13C NMR (DMSO-d6,100 MHz): delta = 210.9, 207.9, 199.3, 169.6, 167.7, 167.6,167.4, 139.7, 138.3, 137.6, 132.8, 130.9, 127.4, 125.1,99.4, 86.0, 83.0, 82.7, 68.9, 76.2, 74.0, 67.4, 66.6, 65.9,57.5, 57.5, 57.4, 56.3, 51.2, 45.6, 43.9, 36.4, 35.6, 35.2,33.8, 32.7, 31.7, 31.3, 30.2, 30.1, 30.0, 28.8, 27.5, 26.9,26.7, 24.9, 22.5, 22.1, 20.8, 16.0, 16.0, 15.1, 14.4, 13.8,10.9; HRESIMS m/z (pos) 1100.4927 [M+ Na]+ (calcd. forC55H84BrNO15 1077.5024).

Reference： [1]Patent: US2007/212371,2007,A1 .Location in patent: Page/Page column 16-17
[2]Medicinal Chemistry Research,2018,vol. 27,p. 583 - 591

5
[ 1392400-31-5 ]
[ 159351-69-6 ]

Yield	Reaction Conditions	Operation in experiment
	With hydrogenchloride; In methanol; water; at 0℃; for 1.5h;	Example 6: 40-O-(2-hydroxyethyl)-rapamycin [everolimus]In a 100 mL flask, 40-O-[2-((2,3-dimethylbut-2-yl)dimethylsilyloxy)ethyl]rapamycin (4.47 g, 4.06 mmol) was dissolved in methanol (20 ml) to give a colorless solution. At 0C, IN aqueous hydrochloric acid (2.0 ml, 2.0 mmol) was added and the mixture was stirred for 90 min. The reaction was followed by TLC (ethyl acetate/n-heptane 3 :2) and HPLC. Then 20 ml of saturated aqueous aHC03 were added, followed by 20 ml of brine and 80 ml of ethyl acetate. The phases were separated and the organic layer was washed with saturated aqueous NaCl until pH 6/7. The organic layer was dried by Na2S04, filtered and concentrated to yield 3.3 g of the product.
	With hydrogenchloride; In methanol; water; at -10℃;Inert atmosphere;	5L reaction bottle, nitrogen protection by adding 0.82kg BP097b03, adding 4.17kg of methanol dissolved, cooled to -10 C, stirring drop 0.82kg 1N-HCl dripping incubation reaction overnight. The HPLC reaction was completed. The reaction droplets were added to 20 kg of water, the solid precipitated, filtered, the filter cake was rinsed with water, and the solid was taken out at -20 C. Preparation of HPLC purification, after treatment to get 0.5kg pure product dissolved in 0.835kg ethanol, drop into 5kg of purified water, precipitation and filtration, solid nitrogen 0 C drying 12h.

Reference： [1]Patent: WO2012/103959,2012,A1 .Location in patent: Page/Page column 18
[2]Patent: CN106478663,2017,A .Location in patent: Paragraph 0023

6
[ 913168-10-2 ]
[ 159351-69-6 ]
[ 1312678-90-2 ]

Yield	Reaction Conditions	Operation in experiment
	With dmap; In dichloromethane; for 0.5h;Inert atmosphere;	In a 10 mL round bottom flask, under argon atmosphere, <strong>[159351-69-6]everolimus</strong> (130 mg, 0.136 mmol) was dissolved in 2.0 mL of CH2CI2. 2-[Benzotriazole-l-yl-(oxycarbonyloxy)-ethyldisulfanyl]- pyridine (104.4 mg, 0.271 mmol) followed by DMAP (49.85 mg, 0.41 mmol) were added. The reaction mixture was stirred for 30 min. Progress of the reaction was monitored by analytical HPLC (0.1% TFA in water, pH = 2.0 and acetonitrile). The reaction mixture was diluted with CH2CI2 and washed with sat. NH4C1. The organic layer was dried over Na2S04 andconcentrated to yield <strong>[159351-69-6]everolimus</strong> (2'-pyridyldisulfanyl)ethyl carbonate, EC0564.

Reference： [1]Patent: WO2011/79227,2011,A1 .Location in patent: Page/Page column 96-97

7
[ 159351-69-6 ]
[ 1422173-29-2 ]

Reference： [1]Journal of the American Chemical Society,2013,vol. 135,p. 2470 - 2473

8
[ 1314705-20-8 ]
[ 159351-69-6 ]

Yield	Reaction Conditions	Operation in experiment
15%	With pyridine; hydrogen fluoride; In tetrahydrofuran; at 0 - 20℃; for 4h;	To a solution of intermediate 17 (280 mg. 0.23 mmol) in THF (10 mL) was added 2 mL HF in pyridine at 0 deg and stirred at rt for 4 hours. Then 20 ml water ws added and extracted with EtOAc (20 mL x 3). The combined organic layer was washed by 0.5N HCI saturated NaHCO3 and brine, dried over anhydrous Na2S04. After concentration the residue was purified with silica gel chromatography (30% to 100 % of EtOAc in petroleum ether as eluent) to give white solid which was further purified by prep-HPLC to give compound A15 (34 mg. 15%) as a white solid.
15%	With pyridine hydrogenfluoride; In tetrahydrofuran; at 0 - 20℃; for 4h;	To a solution of intermediate 17 (280 mg, 0.23 mmol) in THF (10 mL) was added 2 mL HF in pyridine at 0 deg and stirred at rt for 4 hours. Then, 20 mL water was added and extracted with EtOAc (20 mL×3). The combined organic layer was washed by 0.5N HCl, saturated NaHCO3 and brine, dried over anhydrous Na2SO4. After concentration, the residue was purified with silica gel chromatography (30% to 100% of EtOAc in petroleum ether as eluent) to give white solid which was further purified by prep-HPLC to give compound A15 (34 mg, 15%) as a white solid. 1H NMR (300 MHz, CDCl3) delta6.40-6.00 (m, 5H), 5.53-5.25 (m, 4H), 4.83 (s, 1H), 4.13 (m, 1H): LCMS (m/z) ES- 957 (M-1)-.
6.81 g	With pyridine hydrogenfluoride; In tetrahydrofuran; at 0 - 45℃; for 3.5h;	To a solution of 2-((tert-butyldiphenylsilyl)oxy)ethanol (13.1 g, 43.8 mmol) in toluene (51 g) was added N,N-Diisopropylpentan-3-amine (8.7 g, 50.3 mmol) The clear solution was then cooled to 0C and trifluoromethanesulfonic acid anhydride (12.3 g, 43.8 mmol) was added dropwise such that the temperature was maintained between (-2C - 2C). Following the addition a further portion of toluene (5 g) was used for washing. After 1 .5 h N,N-Diisopropylpentan-3-amine (8.7 g, 50.3 mmol) was added followed by toluene (3 g) and Rapamycin (10.0 g, 10.9 mmol ) washing with toluene (18 g). The reaction was then heated to 40C and allowed to stir at this temperature for 22.5 h at which point less than 5 Area% Rapamycin was remaining according to HPLC analysis. The reaction was cooled to ambient temperature and pyridine (1 .0 mL) was then added to quench the reaction which was stirred for a further 30 mins. The reaction was filtered and diluted with isopropyl acetate. The organic solution was washed with 1 M citric acid solution, 10% sodium bicarbonate solution followed by water, dried (MgS04) and concentrated in vacuo. To the crude residue (35.6 g) was added THF (240 mL) and this solution was then added dropwise at 0 C to a HF*pyridine solution (1 :1 , 38.1 g). The reaction was heated to 45 C for 3.5 h then allowed to cool to ambient temperature and diluted with isopropylacetate (300 g). The reaction was then added slowly to an 8% aqueous solution of sodium bicarbonate and further washed with isopropylacetate (250 g). The organic phase was then separated and washed with saturated aqueous sodium chloride solution, dried (MgS04) and concentrated in vacuo. The residue was diluted with isopropylacetate, BHT (0.2% m/m) was added, and the yield of everolimus determined by HPLC analysis against an external standard (6.81 g, 65%).

With hydrogenchloride; In water; acetonitrile; at -10 - 0℃; for 0.5h;

General procedure: Acetonitrile (220 mL) was added to the 500 mL reaction flaskThe compound 7-I-a (21.3 g) obtained in the above step was dissolved with stirring.Controlly add -10 mol/L hydrochloric acid (19.2 ml) at -10 to 0 C.After the reaction was completed for 0.5 h, TLC was monitored until the reaction was completed.Control the temperature at -10 ~ 0 C, add dichloromethane / purified water (220mL / 220mL),Stir layering, the aqueous phase is dichloromethane(106 mL) extraction. Combine the organic phase,Purified water (220 mL × 3), dried over anhydrous sodium sulfate,Filter and add BHT to the filtrate(The ratio of BHT quality to the quality of everolimus 7 is 0.5%),Evaporate at 25 to 30 C under reduced pressure.Get the crude product of everolimus 7-II(19.6g).

Reference： [1]Patent: WO2014/82286,2014,A1 .Location in patent: Paragraph 50
[2]Patent: US2015/51242,2015,A1 .Location in patent: Paragraph 0086
[3]Patent: WO2014/203185,2014,A1 .Location in patent: Page/Page column 22; 23; 24; 26
[4]Patent: CN109206441,2019,A .Location in patent: Paragraph 0081; 0087; 0088; 0089

9
C₇₄H₁₀₁NO₁₆ [ No CAS ]
[ 159351-69-6 ]

Yield	Reaction Conditions	Operation in experiment
621 mg	With 1,1,1,3',3',3'-hexafluoro-propanol; at 50℃; for 1.5h;	Step (a) was performed as above and then the resulting intermediate after chromatography was dissolved in hexafluoroisopropanol (26 mL). The reaction was heated for 1 .5 h at 50 C and then cooled to 0 C and quenched with saturated aqueous sodium bicarbonate solution (26 mL). The organic phase was separated and washed with saturated aqueous sodium chloride solution, water, dried (MgS04) and concentrated. HPLC analysis of the product everolimus against an external standard showed a yield of 621 mg, 31 % over the two steps.

Reference： [1]Patent: WO2014/203185,2014,A1 .Location in patent: Page/Page column 25; 24

10
[ 53123-88-9 ]
[ 159351-69-6 ]

Reference： [1]Patent: WO2014/203185,2014,A1
[2]Patent: WO2016/20664,2016,A1
[3]Patent: CN105254646,2016,A
[4]Patent: CN105254646,2016,A
[5]Patent: CN105254646,2016,A
[6]Patent: CN105254646,2016,A
[7]Patent: WO2016/207205,2016,A1
[8]Patent: CN106146536,2016,A
[9]Patent: CN109776570,2019,A
[10]Patent: CN109776570,2019,A
[11]Patent: CN109776569,2019,A
[12]Patent: CN109776569,2019,A

11
42-O-trityl-everolimus [ No CAS ]
[ 159351-69-6 ]

Yield	Reaction Conditions	Operation in experiment
88%	With hydrogenchloride; In methanol; water;	3 g of the intermediate compound of the formula (2) obtained in Example 3 and 30 ml of methanol were added to a 250 ml capacity flask in an atmospheric state, and the mixture was cooled to a low temperature. 3 ml of 6N HCl was added slowly, and the mixture was stirred for 3 to 4 hours. After completion of the reaction, 100 ml of ethyl acetate and 70 ml of water were added, followed by stirring for 10 minutes. The aqueous layer was separated and removed, and the organic layer was dried with anhydrous magnesium sulfate to remove moisture. The mixture was filtered under reduced pressure, washed with EA, and concentrated under reduced pressure. The product was subjected to a Si60 chromatography column under ethyl acetate to obtain 2.1 g (yield: 88%) of everolimus.
	With 1,1,1,3',3',3'-hexafluoro-propanol; at 58℃; for 3.5h;	Step (a) To a solution of 2-(trityloxy)ethanol (10.7 g, 35.2 mmol) in toluene (42.8 g) was added N,N- Diisopropylpentan-3-amine (7.0 g, 40.4 mmol) The clear solution was then cooled to 0C and Trifluoromethanesulfonic acid anhydride (10.0 g, 35.2 mmol) was added dropwise such that the temperature was maintained between (-2C - 2C). After 2 h N,N-Diisopropylpentan-3-amine (7.0 g, 40.4 mmol) was added to the reaction mixture followed by toluene (14.4 g) and Rapamycin (8.24 g, 8.79 mmol ) washing with toluene (2.4 g). The reaction was then heated to 40 C and allowed to stir at this temperature for 21 h at which point less than 5 Area% Rapamycin was remaining according to HPLC analysis. The reaction was cooled to ambient temperature and pyridine (0.8 ml_) was then added to quench the reaction which was stirred for a further 30 mins. The reaction was diluted with isopropyl acetate. The organic solution was washed with 1 M citric acid solution, 10% sodium bicarbonate solution followed by water, dried (MgS04) and concentrated in vacuo. The reaction mixture was purified using flash column chromatography (ethyl acetate in heptanes) affording a yellow oil (6.85 g, 65% yield). Step (b) The trityl-protected everolimus derivative (5.0 g, 4.165 mmol) was then dissolved in hexafluoroisopropanol and heated to 58 C for 3.5 h. The reaction was then allowed to cool to ambient temperature and was diluted with ethyl acetate (50 ml_) and concentrated in vacuo. This dilution/concentration procedure was repeated once more and then the crude product was filtered over silica gel (25 g) eluting with heptane/ethyl acetate. Recrystallization from heptanes/ethyl acetate afforded the desired product as white crystals (1 .60 g, 40.1 %). HPLC analysis of the mother liquor against an external standard indicated that a further 12% yield Everolimus was contained therein.

Reference： [1]Patent: KR101529963,2015,B1 .Location in patent: Paragraph 0071-0072
[2]Patent: WO2014/203185,2014,A1 .Location in patent: Page/Page column 26; 27

12
[ 159351-68-5 ]
[ 159351-69-6 ]

Yield	Reaction Conditions	Operation in experiment
54%	With triethylamine tris(hydrogen fluoride); In tetrahydrofuran; at 20℃; for 2h;	A solution of TEA3HF (4.65 g, 28.87 mmol) and (27E,29E,31E,32E,36R,37S,38R,39R,40S,43S,44S,45S,47R,48R,57R)-45-[(1R)-2- [(1S,2R,3R)-3-[2-[tert-butyl(dimethyl)silyl]ethoxy]-2-methoxy-cyclohexyl]-1-methyl-ethyl]- 47,57-dihydroxy-44,48-dimethoxy-36,37,38,39,49,50-hexamethyl-67,68-dioxa-59- azatricyclohexatriaconta-27,29,31(49),32(50)-tetraene-51,52,53,54,55-pentone (3.05 g, 2.89 mmol) in THF (50 mL) was stirred at 20 C for 2 hours. The mixture was poured into ice cold saturated NaHCO3 (100mL) then extracted with EtOAc (150 mL × 3). The organic layers were combined and washed with water and brine, then concentrated in vacuo. The residue was purified by reverse phase chromatography (CH3CN/pure water: 7:3) to afford (22E,24E,26E,27E,31R,32S,33R,34R,35S,38S,39S,40S,42R,43R,52R)-42,52-dihydroxy-40- [(1R)-2-[(1S,2R,3R)-3-(2-hydroxyethoxy)-2-methoxy-cyclohexyl]-1-methyl-ethyl]-39,43- dimethoxy-31,32,33,34,44,45-hexamethyl-62,63-dioxa-53-azatricyclohexatriaconta- 22,24,26(44),27(45)-tetraene-46,47,48,49,50-pentone (1.5 g, 54% yield) as a white solid. ESI- MS (EI+, m/z): 980.5 [M+Na]+.
2.96 g	With pyridine hydrogenfluoride; In tetrahydrofuran; at 0 - 45℃; for 1.5h;	To a solution of 2-((tert-butyldimethylsilyl)oxy)ethanol (8.04 g, 43.8 mmol ) in toluene (55 g) was added N,N-diisopropylethylamine (5.94 g, 45.9 mmol ) The clear solution was then cooled to 0C and Trifluoromethanesulfonic acid anhydride (1 1 .97 g, 42.4 mmol) was added dropwise such that the temperature was maintained between (-2C - 2C). Following the addition a further portion of toluene (5 g) was used for washing. After 30 minutes N,N-Diisopropylpentan-3-amine (7.871 g, 45.9 mmol) was added followed by toluene (3 g) and Rapamycin (10.0 g, 10.9 mmol) washing with toluene (18.4 g). The reaction was then heated to 40 C and allowed to stir at this temperature for 42 h at which point less than 5 Area% Rapamycin was remaining according to HPLC analysis. The reaction was cooled to ambient temperature and pyridine (2.6 g) was then added to quench the reaction which was stirred for a further 30 mins. The reaction was filtered and diluted with isopropyl acetate. The organic solution was washed with 1 M citric acid solution, 10% sodium bicarbonate solution followed by water, dried (MgS04) and concentrated in vacuo. The residue was divided into two portions. To half of this crude residue (9.66 g) THF (100 ml_) was added and this solution was then added dropwise at 0 C to a HF*pyridine solution (1 :1 , 17.7 g). A further portion of THF (20 ml_) was used for washing. The reaction was heated to 45 C for 1 .5 h, then allowed to cool to ambient temperature and diluted with isopropylacetate (150 g). The reaction was then added slowly to an 8% aqueous solution of sodium bicarbonate and further washed with isopropylacetate (250 g). The organic phase was then separated and washed with saturated aqueous sodium chloride solution, dried (MgS04) and concentrated in vacuo. The residue was diluted with isopropylacetate, butylhydroxytoluol (BHT; 0.2% m/m) was added, and the yield of everolimus determined by HPLC analysis against an external standard (2.96 g, 57%).
	With hydrogenchloride; In methanol; water; at 0 - 25℃; for 1h;pH 1-3;	Protected everolimus of formula (I la) obtained in step 1 was dissolved in methanol (10 volumes) and chilled to 0-5 C. To this solution was added drop wise, a solution of 1 N HCI. The pH of the reaction was maintained between 1-3. The temperature of the reaction mixture was raised to 25 C and stirred for 1 hour. After completion of reaction, the reaction mixture was diluted with water (15 volumes) and extracted in ethyl acetate (2X20 volumes). The organic layers were combined and washed with brine, dried over sodium sulphate. The organic layer was distilled off under reduced pressure at 30-35 C, to obtain a crude everolimus (0.8 g). The crude everolimus was further purified by preparative HPLC to yield everolimus of purity >99%.

	With hydrogenchloride; In water; acetone; at 20℃; for 0.5h;	Example 2. Synthesis of everolimus The intermediate A (12.98g) was dissolved in 240ml of acetone and was added with 48ml 0.5M hydrochloric acid at 20C and the reacted for 0.5h. Extracted with ethyl acetate, and concentrated to give crude everolimus (12.35g), high performance liquid chromatography analysis showed that the crude product purity greater than 90%.
68%Chromat.	With hydrogenchloride; phosphoric acid; water; In n-heptane; acetonitrile; at 20℃; for 1h;	Example 2: Synthesis of 40-O-(2-hydroxy)ethyl-rapamycin (i.e. everolimus) The crude 40-O-[2-(f-butyldimethylsilyl)oxy]ethyl-rapamycin solution of Example 1 was evaporated to dryness under reduced pressure at 25-30 C. Subsequently, 66 ml of heptane were added to the residue, and the resulting mixture was evaporated to a volume of 40 ml. Then, 48 ml of a (80:20) (v/v) mixture of acetonitrile/water (pH = 1.7, adjusted with 75% ortho-phosphoric acid) were added, and the pH of the resulting mixture was adjusted to 1 .8 with 1 N HCI solution. The resulted mixture was stirred for 1 hour at room temperature, and the lower layer containing everolimus was separated. The overall yield of everolimus as determined by HPLC using an external standard was 68% (71 1 mg).
	With hydrogenchloride; In water; acetonitrile; at -10 - 0℃; for 0.5h;	Acetonitrile (220 mL) was added to the 500 mL reaction flaskThe compound 7-I-a (21.3 g) obtained in the above step was dissolved with stirring.Controlly add -10 mol/L hydrochloric acid (19.2 ml) at -10 to 0 C.After the reaction was completed for 0.5 h, TLC was monitored until the reaction was completed.Control the temperature at -10 ~ 0 C, add dichloromethane / purified water (220mL / 220mL),Stir layering, the aqueous phase is dichloromethane(106 mL) extraction. Combine the organic phase,Purified water (220 mL × 3), dried over anhydrous sodium sulfate,Filter and add BHT to the filtrate(The ratio of BHT quality to the quality of everolimus 7 is 0.5%),Evaporate at 25 to 30 C under reduced pressure.Get the crude product of everolimus 7-II(19.6g).

Reference： [1]Patent: WO2019/241789,2019,A1 .Location in patent: Paragraph 00349; 00352
[2]Patent: WO2014/203185,2014,A1 .Location in patent: Page/Page column 21; 22
[3]Patent: WO2016/20664,2016,A1 .Location in patent: Page/Page column 22
[4]Patent: CN103848849,2016,B .Location in patent: Paragraph 0027; 0030; 0031
[5]Patent: WO2016/207205,2016,A1 .Location in patent: Page/Page column 13
[6]Patent: CN109206441,2019,A .Location in patent: Paragraph 0077; 0078; 0081

13
[ 141-82-2 ]
[ 159351-69-6 ]
C₅₄H₈₃NO₁₅ [ No CAS ]

Yield	Reaction Conditions	Operation in experiment
	In methanol; at 20℃; for 10h;	A compound represented by Chemical Formula 4 above was synthesized by mixing <strong>[159351-69-6]Everolimus</strong> (LC Laboratories) as a starting material and malonic acid (Sigma-Aldrich) at a molar ratio of 1:1 to 2 and reacting and stirring the mixture in a methanol solvent at room temperature for 10 hours.

Reference： [1]Patent: US2015/368297,2015,A1 .Location in patent: Paragraph 0045

14
42-O-[2-(methoxymethoxy)ethyl]rapamycin [ No CAS ]
[ 159351-69-6 ]

Yield	Reaction Conditions	Operation in experiment
55%	With hydrogenchloride; In methanol; water; at 5℃;	The resulting mixture of 42-O- [2 (methoxymethoxy) ethyl] rapamycin(1.20 g, 1.20 mmol) was dissolved10 mL of methanol, the ice bath so that the solution temperature dropped to below 5 C, slowly dropping lwt% hydrochloric acid aqueous solution to the reaction system pH value of 2 to 3; slowly heated to 20 C, continue stirring until the liquid (20 mL x 3). The organic layers were combined and washed with saturated brine (20 mL x 2). The organic layer was washed with water, dried over anhydrous magnesium sulfate, and evaporated to dryness under reduced pressure. The reaction mixture was cooled to 0 C, , Dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The obtained crude product was subjected to silica gel column chromatography, eluting with ethyl acetate to obtain 632 mg of the product of everolimus. The HPLC purity was 99% and the yield was 55%.

Reference： [1]Patent: CN105254646,2016,A .Location in patent: Paragraph 0025; 0029; 0030

15
42-O-[2(tetrahydro-2H-furan-2-oxy)ethyl]rapamycin [ No CAS ]
[ 159351-69-6 ]

Yield	Reaction Conditions	Operation in experiment
61%	With hydrogenchloride; In methanol; water; at 5℃;	The resulting mixture of 42-O- [2 (tetrahydro-2H-furan-2-oxy) ethyl] rapamycin(1.10 g,1. 06 mmol) was dissolved in 10 mL of methanol, and the temperature of the solution was reduced to below 5 C in an ice-water bath. The aqueous solution of 1 wt% hydrochloric acid was slowly added dropwise to a pH of 2 to 3 in the reaction system; And the mixture was cooled to 0 C. Saturated sodium bicarbonate solution was added dropwise to pH 7-8 and extracted with ethyl acetate (20 mL X 3). The organic layers were combined and washed with saturated salt Washed with water (20 mL X2), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The obtained crude product was subjected to silica gel column chromatography, eluting with ethyl acetate to obtain 617 mg of everolimus. The HPLC purity was 99% and the yield was 61%.

Reference： [1]Patent: CN105254646,2016,A .Location in patent: Paragraph 0032; 0036; 0037

16
42-O-[2(tetrahydrofuran-2-oxy)ethyl]rapamycin [ No CAS ]
[ 159351-69-6 ]

Yield	Reaction Conditions	Operation in experiment
67%	With hydrogenchloride; In methanol; water; at 5℃;	The resulting mixture of 42-O- [2 (tetrahydrofuran-2-oxy) ethyl] rapamycin(1.20 g, 1.17 mmol)Dissolved in 10mL of methanol, the ice bath so that the solution temperature dropped to below 5 C, slowly dropping lwt% hydrochloric acid aqueous solution to the reaction system in the pH value of 3 to 4; slowly heated to 20 C, (20 mL x 3). The organic layers were combined and washed with saturated brine (20 mL x 20 mL x 3). The reaction mixture was cooled to 0 C and the solution was extracted with ethyl acetate 2), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude product was subjected to silica gel column chromatography using ethyl acetate as eluant to give the product of everolimus 749 mg. The HPLC purity was 98% and the molar yield was 67%.

Reference： [1]Patent: CN105254646,2016,A .Location in patent: Paragraph 0039; 0043; 0044

17
42-O-[2-(1-ethoxyethoxy)ethyl]rapamycin [ No CAS ]
[ 159351-69-6 ]

Yield	Reaction Conditions	Operation in experiment
65%	With hydrogenchloride; In methanol; water; at 5℃;	The resulting 42-0- [2- (I-ethoxyethoxy) -ethyl] -sapamycin was obtained in the previous step(1. 05 g, 1.02 mmol)Dissolved in 10mL of methanol, the ice bath so that the solution temperature dropped to below 5 C, slowly dropping lwt% hydrochloric acid aqueous solution to the reaction system in the pH value of 3 to 4; slowly heated to 20 C, (20 mL x 3). The organic layers were combined and washed with saturated brine (20 mL x 20 mL x 3). The reaction mixture was cooled to 0 C and the solution was extracted with ethyl acetate 2), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude product was subjected to silica gel column chromatography using ethyl acetate as eluant to give 635 mg of the product of everolimus. The HPLC purity was 98% and the molar yield was 65%.

Reference： [1]Patent: CN105254646,2016,A .Location in patent: Paragraph 0045; 0049; 0050

18
42-O-[2-(1-methyl-1-methoxyethoxy)ethyl]rapamycin [ No CAS ]
[ 159351-69-6 ]

Yield	Reaction Conditions	Operation in experiment
68%	With hydrogenchloride; In methanol; water; at 5℃;pH 3 - 4;	The resulting mixture of 42-O- [2- (1-methyl-1-methoxyethoxy) ethyl] rapamycin(1. 05 g,1. 02mmol) was dissolved in 10mL methanol. The temperature of the solution was reduced to below 5 C in an ice-water bath. The aqueous solution of 1wt% hydrochloric acid was slowly added dropwise to a pH value of 3 to 4 in the reaction system; And the mixture was cooled to 0 C. Saturated sodium bicarbonate solution was added dropwise to pH 7-8 and extracted with ethyl acetate (20 mL X 3). The combined organic layer was washed with saturated salt Washed with water (20 mL X2), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The obtained crude product was subjected to silica gel column chromatography, eluting with ethyl acetate, to obtain the product of everolimus 664 mg, HPLC purity of 99% and molar yield of 68%.

Reference： [1]Patent: CN105254646,2016,A .Location in patent: Paragraph 0052; 0056; 0057

19
40-O-[2-(triisopropylsilyl)oxy]ethyl-rapamycin [ No CAS ]
[ 159351-69-6 ]

Yield	Reaction Conditions	Operation in experiment
64%Chromat.	With hydrogenchloride; phosphoric acid; water; In n-heptane; acetonitrile; at 20℃; for 24h;	Example 4: Synthesis of 40-O-(2-hydroxy)ethyl-rapamycin (i.e. everolimus) The crude 40-O-[2-(triisopropylsilyl)oxy]ethyl-rapamycin solution of Example 3 was evaporated to dryness under reduced pressure at 25-30 C. Subsequently, 66 ml of heptane were added to the residue, and the resulting mixture was evaporated to a volume of 40 ml. Then, 48 ml of a (80:20) (v/v) mixture of acetonitrile/water (pH = 1.7, adjusted with 75% ortho-phosphoric acid) were added, and the pH of the resulting mixture was adjusted to 1 .8 with 1 N HCI solution. The resulted mixture was stirred 1 day at room temperature, and the lower layer containing everolimus was separated. The overall yield of everolimus as determined by HPLC using an external standard was 64% (668 mg).

Reference： [1]Patent: WO2016/207205,2016,A1 .Location in patent: Page/Page column 14

20
[ 331810-13-0 ]
[ 159351-69-6 ]

Reference： [1]Patent: CN106146536,2016,A
[2]Patent: CN109776570,2019,A
[3]Patent: CN109776569,2019,A

21
C₅₆H₉₁NO₁₄Si [ No CAS ]
[ 159351-69-6 ]

Yield	Reaction Conditions	Operation in experiment
3.8 g	With hydrogenchloride; In methanol; water; for 4.5h;Cooling with ice;	6.8 g of the coupling product was dissolved in a 1 L single-necked flask with 500 ml of methanol, cooled in an ice-water bath, and 10 ml of HCl was added dropwise over half an hour. After completion of the dropwise addition, the reaction was carried out for 4 hours. After completion of the reaction, the residue was evaporated to dryness under reduced pressure, and the residue was dissolved with 500 ml of ethyl acetate, washed with saturated brine and extracted with ethyl acetate. The organic layers were combined, dried over anhydrous sodium sulfate, evaporated to dryness under reduced pressure to give a yellow oil which was separated by column to give the crude product 5.6g.;_Example 4: Preparation of everolimus 5 grams of everolimus crude, 200ml of acetonitrile and 120ml of purified water dissolved, prepared into a sample solution, filtered through the 0.45 micron filter, to be injected; Start the injection pump to 500ml / min flow rate of injection for 1.4min, and then open the gradient elution program began to elute, to the target peak appears, adjust the switching valve to collect the target peak, sampling by HPLC detection, other parts of the target peak include isomer peaks are collected into the nonconforming container and then switched to the waste collection tank; The above procedure was repeated until the batch was completely finished. The collected liquid was distilled under reduced pressure at room temperature with a rotary evaporator to obtain 3.8 g of the everolimus powdery solid.

Reference： [1]Patent: CN106146536,2016,A .Location in patent: Paragraph 0020; 0022; 0032; 0033; 0034; 0035

22
[ 75-21-8 ]
[ 53123-88-9 ]
[ 159351-69-6 ]

Yield	Reaction Conditions	Operation in experiment
6.9 g	With trifluorormethanesulfonic acid; In toluene; at 70℃; under 7500.75 Torr; for 8h;Autoclave;	9.14 g rapamycin (0.01mol), 11g ethylene oxide (0.25 mol), 0.1 g trifluoromethanesulfonic acid, 30mL toluene was added to the autoclave after mixing, Warmed to 70 C, Maintaining 1.0 MPa pressure, the reaction was stopped after 8 hours, cooled to room temperature,The solvent was recovered under reduced pressure and the residue was purified by silica gel column chromatography (200-300 mesh silica gel,Eluent: ethyl acetate: petroleum ether = 20: 1),7.7 g 97.95% of everolimus was obtained, which was purified by HP-20 resin column chromatography (eluent: acetonitrile: water = 65:35) 6.9 g of everolimus was obtained as a white solid, HPLC purity:99.6%, Isomer content:0.12%. Molar yield:72.0%

Reference： [1]Patent: CN104876944,2017,B .Location in patent: Paragraph 0035-0050

23
[ 159351-69-6 ]
C₅₃H₈₃NO₁₄ [ No CAS ]

Yield	Reaction Conditions	Operation in experiment
45%	With zinc(II) chloride; In tetrahydrofuran; at 60℃; for 2h;	<strong>[159351-69-6]Everolimus</strong> (0.2 g) was dissolved in THF (15 ml)ZnCl2 (0.3 g) was added with stirring, heated to 60 C,The reaction for 2 hours. The reaction solution was poured into water (200 ml), stirred for 10 minutes,The mixture was extracted with ethyl acetate (150 ml × 3). The organic phases were combined,Washed twice with saturated saline, dried over anhydrous Na2SO4, filtered, and the filtrate was concentrated to dryness under vacuum at 30 C.The crude product was purified by column chromatography (n-hexane: ethyl acetate = 1: 2) to give the title product (90 mg) in 45% yield.

Reference： [1]Patent: CN104557975,2017,B .Location in patent: Paragraph 0035; 0036; 0037; 0038; 0039; 0040; 0041; 0042

24
[ 107-21-1 ]
[ 53123-88-9 ]
[ 159351-69-6 ]

Yield	Reaction Conditions	Operation in experiment
5.7 kg		620g of ethylene glycol and 6L of tetrahydrofuran were added to the reaction flask,Mix well to mix.The reaction temperature was controlled at 10 .Under nitrogen protection,1.41L trifluoroacetic anhydride was slowly added dropwise,Dropping is completed,Reaction for 1.5 hours,The reaction solution.9.14 kg of rapamycin was dissolved in 54 L of tetrahydrofuran,Added to the reaction solution,The reaction temperature was controlled at 10 .Slowly add 13ml boron trifluoride diethyl ether solution. Bi completed,The reaction was stirred for 2 hours. After the reaction is completed,60L saturated aqueous sodium bicarbonate solution was added,Stir wellThen suction filtered,To the filtrate was added 30 L of ethyl acetate,Liquid separation,The organic phase is washed with pure water until nearly neutral.The organic phase was dried over 500 g of anhydrous sodium sulfate for 2 hours, filtered,Concentrated under reduced pressure to a solventless outflow,A thick liquid. Column chromatography,The eluent is petroleum ether:Ethyl acetate = 1: 6. The collected effluent was concentrated under reduced pressure to give 6.3 kg of yellow foamy solid,Yield 66%.A mixture of 26.8 L of methanol and ethyl acetate (v / v = 1/3) was added to the above yellow foamy solid,Stirring to dissolve,The temperature was controlled at 25 for 30 minutes,13.4 L cyclohexane was added dropwise,Bi completed,The temperature was controlled at 12 for 2 hours,Cool the feed liquid to about 0 slowly stirring 3h,Suction filtration,Drying at room temperature under vacuum gave 5.7 kg of a white solid,HPLC and mass spectrometry determined that the white solid was everolimus,Purity 98.1%.

Reference： [1]Patent: CN106146535,2016,A .Location in patent: Paragraph 0037; 0038; 0039; 0040; 0041; 0042; 0043-0056

25
[ 2549-51-1 ]
[ 159351-69-6 ]
C₅₅H₈₄ClNO₁₅ [ No CAS ]

Yield	Reaction Conditions	Operation in experiment
0.958 g	In dichloromethane; at 20℃; for 4h;Enzymatic reaction;	<strong>[159351-69-6]Everolimus</strong> (1.0 g, 1.04 mmol) and vinyl chloroacetate(0.188 g, 1.56 mmol) were dissolved in dichloromethane(10 mL). Then the immobilized lipase (0.5 g) was added tothe mixture, which was stirred for about 4 h at room temperature.After reaction finished, the immobilized lipase wasfiltered with a funnel and the filtrate was concentrated invacuo. The residue was purified by silica gel columnchromatography (n-hexane: acetone = 5:1 to 3:1) to obtaincompound 1 (0.958 g, 0.93 mmol) as a white powder. Mp70.8-71.3 C; [alpha]20D -129.6 (c 1.0, MeOH); infrared (IR)(KBr) numax 3425, 2932, 2871, 1721, 1643 cm-1; 1H NMR(dimethyl sulphoxide (DMSO)-d6, 400 MHz): delta = 6.46 (1H,d, J = 13.6 Hz), 6.40 (1H, d, J = 11.2 Hz), 6.23 (1H, t, J =13.4 Hz), 6.12 (1H, d, J = 10.6 Hz), 5.49-5.43 (1H, m),5.10 (1H, d, J = 9.2 Hz), 4.94 (1H, br s), 4.37 (2H, br s),4.26-4.20 (2H, m), 4.01-3.96 (2H, m), 3.72 (2H, br s), 3.64(1H, d, J = 11.0 Hz), 3.42 (2H, d, J = 11.0 Hz), 3.15 (3H,br s), 3.05 (3H, br s), 3.01-3.05 (2H, m), 2.71-2.75 (2H,m), 2.49-2.48 (1H, m), 2.35-2.38 (2H, m), 2.22-2.25 (2H,m), 2.11 (2H, br s), 1.99-2.02 (2H, m), 1.85-1.91 (3H, m),1.74 (2H, br s), 1.63 (6H, br s), 1.55-1.60 (6H, m),1.35-1.40 (2H, m), 1.20-1.25 (4H, m), 1.14 (4H, br s),0.97-1.04 (6H, m), 0.83-0.86 (6H, m), 0.73-0.78 (6H, m);13C NMR (DMSO-d6, 100 MHz): delta = 210.9, 208.7, 207.9,198.9, 169.6, 169.0, 167.7, 167.4, 139.7, 138.3, 137.6,132.8, 130.9, 127.4, 125.4, 99.4, 86.0, 83.0, 82.9, 82.7,76.2, 74.0, 68.9, 67.4, 66.6, 65.8, 57.5, 57.4, 55.9, 55.3,51.2, 45.6, 43.9, 41.9, 41.5, 38.7, 36.4, 35.6, 35.2, 33.8,32.7, 32.5, 31.3, 30.2, 30.0, 26.9, 26.7, 24.9, 22.1, 20.8, 16.0, 15.1, 13.8, 13.8, 10.9; HRESIMS m/z (pos) 1056.5414[M + Na]+ (calcd. for C55H84ClNO15 1033.5529).

Reference： [1]Medicinal Chemistry Research,2018,vol. 27,p. 583 - 591

26
28-O-TMS-42-O-TBS everolimus [ No CAS ]
[ 159351-69-6 ]

Yield	Reaction Conditions	Operation in experiment
68.3%	With hydrogenchloride; In water; acetone; at 0 - 5℃; for 1h;	28-O-TMS-42-O-TBS everolimus (intermediate C-1, 7.84 g) was dissolved in 200 ml of acetone, 1M hydrochloric acid (40 ml) was added, and reacted at 0-5 C for 1 h, then ethyl acetate After extraction and concentration, the crude everolimus (6.98 g) was obtained as a pale yellow foamy solid.High performance liquid chromatography showed a crude product purity of 76%.The crude product was purified to obtain 3.58 g of the finished product of everolimus.It was a white solid with an HPLC purity of 97.2% and a total yield of 68.3% from rapamycin.
20.4 g	With hydrogenchloride; In water; acetonitrile; at -10 - 0℃; for 0.5h;	Acetonitrile (220 mL) and Compound 6 (22 g) were sequentially added to a 500 mL reaction flask.Stir and dissolve.Control the temperature slowly add 0.2mol/L at -10~0 CHydrochloric acid (19.2 ml),After the reaction was completed for 0.5 h, TLC was monitored until the reaction was completed.Controlled temperature was added to dichloromethane/purified water (220 mL/220 mL) at -10 to 0 C.The layers were stirred and the aqueous extracted with dichloromethane (EtOAc)The organic phases were combined and washed with purified water (220 mL×3).Dry over anhydrous sodium sulfate, filter,Concentrated to dry solids of everolimus 7(20.4g).
3.58 g	With hydrogenchloride; In acetone; at 0 - 5℃; for 1h;	28-O-TMS-42-O-TBS Everolimus (Intermediate C-1, 7.84g)Dissolved in 200 ml of acetone, added with 1 M hydrochloric acid (40 ml), and reacted at 0-5 C for 1 h.Extracted with ethyl acetate and concentrated to give crude everolimus (6.98 g).As a pale yellow foamy solid, high performance liquid chromatography showed a crude product purity of 76%.After the crude product was purified, 3.58 g of the finished product of everolimus was obtained as a white solid.The HPLC purity was 97.2%, and the total yield from rapamycin was 68.3%.

Reference： [1]Patent: CN109776570,2019,A .Location in patent: Paragraph 0062; 0070; 0071; 0072
[2]Patent: CN109206441,2019,A .Location in patent: Paragraph 0074; 0075; 0076
[3]Patent: CN109776569,2019,A .Location in patent: Paragraph 0059-0061; 0069-0071

27
[ 18162-48-6 ]
[ 159351-69-6 ]
[ 159351-68-5 ]

Yield	Reaction Conditions	Operation in experiment
21.3 g	With 1H-imidazole; In dichloromethane; at -10 - 0℃; for 0.333333h;	Where R is TBS.The above concentrate was added to the 500 mL reaction flask in sequence.(<strong>[159351-69-6]everolimus</strong> 7, 20.4 g) and dichloromethane (212 mL),The mixture was stirred and dissolved, and imidazole (2.36 g) was added and stirred to dissolve.Slowly add TBS-Cl (5.21g) at a temperature of -10 to 0 C.a solution of dichloromethane (15 mL), after completion, the reaction was carried out for 20 min.The TLC was monitored until the reaction was completed. Add purified water (212 mL),The layers were stirred and the organic phase was washed with purified water (238 mL×2).Dry over anhydrous sodium sulfate, filter, pass silica gel column(ethyl acetate/n-hexane = 1/1), collecting the desired components,Concentrated to dry at 25-30 C,Compound 7-I-a (21.3 g) was obtained.

Reference： [1]Patent: CN109206441,2019,A .Location in patent: Paragraph 0077; 0078; 0079; 0080

28
C₅₆H₉₁NO₁₄Si [ No CAS ]
[ 159351-69-6 ]

Yield	Reaction Conditions	Operation in experiment
	With hydrogenchloride; In water; acetonitrile; at -10 - 0℃; for 0.5h;	General procedure: Acetonitrile (220 mL) was added to the 500 mL reaction flaskThe compound 7-I-a (21.3 g) obtained in the above step was dissolved with stirring.Controlly add -10 mol/L hydrochloric acid (19.2 ml) at -10 to 0 C.After the reaction was completed for 0.5 h, TLC was monitored until the reaction was completed.Control the temperature at -10 ~ 0 C, add dichloromethane / purified water (220mL / 220mL),Stir layering, the aqueous phase is dichloromethane(106 mL) extraction. Combine the organic phase,Purified water (220 mL × 3), dried over anhydrous sodium sulfate,Filter and add BHT to the filtrate(The ratio of BHT quality to the quality of everolimus 7 is 0.5%),Evaporate at 25 to 30 C under reduced pressure.Get the crude product of everolimus 7-II(19.6g).

Reference： [1]Patent: CN109206441,2019,A .Location in patent: Paragraph 0081; 0084; 0085; 0086

29
C₅₉H₉₇NO₁₄Si [ No CAS ]
[ 159351-69-6 ]

Yield	Reaction Conditions	Operation in experiment
	With hydrogenchloride; In water; acetonitrile; at -10 - 0℃; for 0.5h;	General procedure: Acetonitrile (220 mL) was added to the 500 mL reaction flaskThe compound 7-I-a (21.3 g) obtained in the above step was dissolved with stirring.Controlly add -10 mol/L hydrochloric acid (19.2 ml) at -10 to 0 C.After the reaction was completed for 0.5 h, TLC was monitored until the reaction was completed.Control the temperature at -10 ~ 0 C, add dichloromethane / purified water (220mL / 220mL),Stir layering, the aqueous phase is dichloromethane(106 mL) extraction. Combine the organic phase,Purified water (220 mL × 3), dried over anhydrous sodium sulfate,Filter and add BHT to the filtrate(The ratio of BHT quality to the quality of everolimus 7 is 0.5%),Evaporate at 25 to 30 C under reduced pressure.Get the crude product of everolimus 7-II(19.6g).

Reference： [1]Patent: CN109206441,2019,A .Location in patent: Paragraph 0081; 0087; 0088; 0089

30
[ 58479-61-1 ]
[ 159351-69-6 ]
[ 1314705-20-8 ]

Yield	Reaction Conditions	Operation in experiment
21.3 g	With 1H-imidazole; In dichloromethane; at -10 - 0℃; for 0.333333h;	Take <strong>[159351-69-6]everolimus</strong> 7 (20.4 g) (prepared as described in Example 3) and dichloromethane (200 ml),Add to the reaction flask, stir to dissolve, add imidazole (2.3g),Stir and dissolve. Slowly add TBDPS-Cl/dichloromethane at -10 to 0 C(9.4g/25ml) solution, after the reaction was completed for 20min, TLC monitoring was started.The basic reaction to <strong>[159351-69-6]everolimus</strong> is completed. Add 200 mL of purified water,The layers were stirred and the organic phase was washed with purified water (230 mL×2 times).Dry over anhydrous sodium sulfate for more than 0.5 h, filter, and pass through a silica gel column.The desired fractions were collected and concentrated to dryness at 25-30 C.The above concentrate was subjected to the method of Example 4 to remove the silicon-based protecting group.Further referring to the method of Embodiment 5,Prepare column on C18 (filler type: C18(II)-EP-DM-10-110A,Mobile phase (V:V): acetonitrile: water = 50:50 (containing 0.1% acetic acid)),The liquid phase purified <strong>[159351-69-6]everolimus</strong> 7-II was prepared.

Reference： [1]Patent: CN109206441,2019,A .Location in patent: Paragraph 0087; 0088; 0089

31
[ 994-30-9 ]
[ 159351-69-6 ]
C₅₉H₉₇NO₁₄Si [ No CAS ]

Yield	Reaction Conditions	Operation in experiment
21.3 g	With pyridine; In dichloromethane; at -10 - 0℃; for 0.333333h;	Take <strong>[159351-69-6]everolimus</strong> 7 (20.4 g) (prepared as described in Example 3) and dichloromethane (200 ml),The reaction mixture was placed in a reaction flask, stirred and dissolved, and pyridine (2.7 g) was added thereto, followed by stirring to dissolve.TAS-Cl/dichloromethane (5.1 g/20 ml) solution was slowly added dropwise at -10 to 0 C.After the reaction was completed for 20 min, TLC monitoring was started until the basic reaction of <strong>[159351-69-6]everolimus</strong> was completed.Add 200 mL of purified water and stir the layers.The organic phase was washed with purified water (215 mL x 2 times).Dry over anhydrous sodium sulfate for more than 0.5 h, filter, and pass through a silica gel column.The desired fractions were collected and concentrated to dryness at 25-30 C.The above-mentioned concentrate was subjected to the method of Example 4 to remove the silicon-based protective group, and further referred to the method of Example 5, and the system was prepared by C18.Spare column (filler type: C18(II)-EP-DM-10-110A,Mobile phase (V:V): acetonitrile: water = 50:50 (containing 0.1% acetic acid)),The liquid phase purified <strong>[159351-69-6]everolimus</strong> 7-II was prepared.

Reference： [1]Patent: CN109206441,2019,A .Location in patent: Paragraph 0087; 0088; 0089

32
[ 75-77-4 ]
[ 159351-69-6 ]
C₅₆H₉₁NO₁₄Si [ No CAS ]

Yield	Reaction Conditions	Operation in experiment
21.3 g	With triethylamine; In dichloromethane; at -10 - 0℃; for 0.333333h;	Take <strong>[159351-69-6]everolimus</strong> 7 (20.4 g) (prepared as described in Example 3) and dichloromethane (200 ml),Add to the reaction flask, stir to dissolve,Triethylamine (3.4 g) was added and dissolved by stirring.Slowly add TMS-Cl/dichloromethane at -10 to 0 C(3.7 g/10 ml) solution, starting TLC monitoring, until the basic reaction of <strong>[159351-69-6]everolimus</strong> was completed.Add 200 mL of purified water and stir the layers.The organic phase was washed with purified water (220 mL x 2 times).Dry over anhydrous sodium sulfate for more than 0.5 h, filter, and pass through a silica gel column.Collect the required components,Concentrate to dryness at 25-30 C.The above concentrate was subjected to the method of Example 4 to remove the silicon-based protecting group.Further referring to the method of Example 5, the column was prepared on C18 (filler type: C18(II)-EP-DM-10-110A, mobile phase (V:V): acetonitrile:Water = 50:50 (containing 0.1% acetic acid)),The liquid phase purified <strong>[159351-69-6]everolimus</strong> 7-II was prepared.

Reference： [1]Patent: CN109206441,2019,A .Location in patent: Paragraph 0084; 0085; 0086

33
28-O-TES-42-O-THDMS everolimus [ No CAS ]
[ 159351-69-6 ]

Yield	Reaction Conditions	Operation in experiment
71%	With sulfuric acid; In tetrahydrofuran; at 0 - 10℃; for 1h;	28-O-TES-42-O-THDMS Everolimus Pure (Intermediate C-2, 8.23 g) was dissolved in 200 ml of tetrahydrofuran, 1M dilute sulfuric acid (50 ml) was added, and reacted at 0-10 C for 1 h. The mixture was extracted with ethyl acetate and concentrated to give a crude product (7.52 g) as a yellow solid. The crude product was purified to obtain 3.72 g of the finished product of everolimus, which was an off-white solid with HPLC purity of 96.3%, and the total yield from rapamycin was 71.0%.
3.72 g	With sulfuric acid; In tetrahydrofuran; at 0 - 10℃; for 1h;	28-O-TES-42-O-THDMS Everolimus Pure (Intermediate C-2, 8.23g)Dissolved in 200 ml of tetrahydrofuran, added with 1 M dilute sulfuric acid (50 ml), and reacted at 0-10 C for 1 h.Extracted with ethyl acetate and concentrated to give a crude (1.High performance liquid chromatography showed a crude product purity of 73%.After the crude product was purified, 3.72 g of the finished product of everolimus was obtained as an off-white solid.The HPLC purity was 96.3%, and the total yield from rapamycin was 71.0%.

Reference： [1]Patent: CN109776570,2019,A .Location in patent: Paragraph 0073; 0074; 0075; 0083; 0084; 0085
[2]Patent: CN109776569,2019,A .Location in patent: Paragraph 0072-0074; 0082-0084

34
[ 331810-10-7 ]
[ 159351-69-6 ]

Reference： [1]Patent: CN109776570,2019,A
[2]Patent: CN109776569,2019,A

35
[ 155435-45-3 ]
[ 159351-69-6 ]

Reference： [1]Patent: CN109776570,2019,A
[2]Patent: CN109776569,2019,A

36
[ 13296-57-6 ]
[ 159351-69-6 ]
2-[[(22E,24E,26E,27E,33R,34S,35R,36R,38S,40S,42R,43S,44R,45R,55R)-44,55-dihydroxy-43-[(1R)-2-[(1S,3R,4R)-4-(2-hydroxyethoxy)-3-methoxy-cyclohexyl]-1-methyl-ethyl]-45-methoxy-33,34,35,36,46,47-hexamethyl-48,49,50,51,52-pentaoxo-67,68-dioxa-57-azatricyclohexatriaconta-22,24,26⁽⁴⁶⁾,27⁽⁴⁷⁾-tetraen-42-yl]oxy]ethyl N-methylcarbamate [ No CAS ]
2-[[(22E,24E,26E,27E,33R,34S,35R,36R,38S,40S,42S,43S,44R,45R,55R)-44,55-dihydroxy-43-[(1R)-2-[(1S,3R,4R)-4-(2-hydroxyethoxy)-3-methoxy-cyclohexyl]-1-methyl-ethyl]-45-methoxy-33,34,35,36,46,47-hexamethyl-48,49,50,51,52-pentaoxo-67,68-dioxa-57-azatricyclohexatriaconta-22,24,26⁽⁴⁶⁾,27⁽⁴⁷⁾-tetraen-42-yl]oxy]ethyl N-methylcarbamate [ No CAS ]

Yield	Reaction Conditions	Operation in experiment
20 mg; 18 mg	With toluene-4-sulfonic acid; In tetrahydrofuran; at 0 - 23℃; for 3.5h;Inert atmosphere;	To a degassed solution of <strong>[159351-69-6]everolimus</strong> (0.5 g, 0.52 mmol) in THF (10 mL) at 0C, was added p-toluenesulfonic acid (0.45 g, 2.61 mmol) and 2-hydroxyethyl N-methylcarbamate (2.80 mL). The resulting mixture was stirred at 0 C for 0.5 h under N2, then warmed 23C and stirred for 3h. The mixture was poured into sat.NaHCO3 (40 mL) which was extracted with EtOAc (30 mL). The organic layer was washed with water (30 mL × 2), brine (40 mL), dried over anhydrous Na2SO4, filtered, and concentrated. The residue was purified by silica gel chromatography (DCM:MeOH = 10:1) then additionally purified via reverse phase chromatography (C18, CH3CN:H2O = 7:3) to provide 2- [[(22E,24E,26E,27E,33R,34S,35R,36R,38S,40S,43S,44R,45R,55R)-44,55-dihydroxy-43- [(1R)-2-[(1S,3R,4R)-4-(2-hydroxyethoxy)-3-methoxy-cyclohexyl]-1-methyl-ethyl]-45- methoxy-33,34,35,36,46,47-hexamethyl-48,49,50,51,52-pentaoxo-67,68-dioxa-57- azatricyclohexatriaconta-22,24,26(46),27(47)-tetraen-42-yl]oxy]ethyl N-methylcarbamate (I- 81: 100 mg, 18% yield) as a white solid. ESI-MS (EI+, m/z): 1067.4 [M+Na]+. 1H NMR (400 MHz, CDCl3) d 6.50-5.91 (m, 4H), 5.58- 4.97 (m, 4H), 4.70 (s, 1H), 4.51 (d, J = 40.4 Hz, 1H), 4.33- 4.02 (m, 3H), 3.93- 3.62 (m, 6H), 3.61- 3.00 (m, 13H), 2.86- 2.46 (m, 6H), 2.40 - 2.22 (m, 2H), 2.18-1.69 (m, 22H), 1.58-1.25 (m, 7H), 1.24- 0.79 (m, 18H), 0.79- 0.62 (m, 1H). 120 mg of 2-[[(22E,24E,26E,27E,33R,34S,35R,36R,38S,40S,43S,44R,45R,55R)- 44,55-dihydroxy-43-[(1R)-2-[(1S,3R,4R)-4-(2-hydroxyethoxy)-3-methoxy-cyclohexyl]-1- methyl-ethyl]-45-methoxy-33,34,35,36,46,47-hexamethyl-48,49,50,51,52-pentaoxo-67,68- dioxa-57-azatricyclohexatriaconta-22,24,26(46),27(47)-tetraen-42-yl]oxy]ethyl N- methylcarbamate was purified via prep chiral HPLC and the resulting epimers purified via silica gel chromatography (hexane:DCM:EtOAc:MeOH= 3:3:1:0.6) to provide 2- [[(22E,24E,26E,27E,33R,34S,35R,36R,38S,40S,42S,43S,44R,45R,55R)-44,55-dihydroxy- 43-[(1R)-2-[(1S,3R,4R)-4-(2-hydroxyethoxy)-3-methoxy-cyclohexyl]-1-methyl-ethyl]-45- methoxy-33,34,35,36,46,47-hexamethyl-48,49,50,51,52-pentaoxo-67,68-dioxa-57- azatricyclohexatriaconta-22,24,26(46),27(47)-tetraen-42-yl]oxy]ethyl N-methylcarbamate (I- 75: 18 mg, 15% yield) and 2- [[(22E,24E,26E,27E,33R,34S,35R,36R,38S,40S,42R,43S,44R,45R,55R)-44,55-dihydroxy- 43-[(1R)-2-[(1S,3R,4R)-4-(2-hydroxyethoxy)-3-methoxy-cyclohexyl]-1-methyl-ethyl]-45- methoxy-33,34,35,36,46,47-hexamethyl-48,49,50,51,52-pentaoxo-67,68-dioxa-57- azatricyclohexatriaconta-22,24,26(46),27(47)-tetraen-42-yl]oxy]ethyl N-methylcarbamate (I- 74: 20 mg, 16% yield), both as white solids. Chiral separation method: I-75: ESI-MS (EI+, m/z): 1067.3 [M+Na]+. 1H NMR (400 MHz, CDCl3) d 6.33 (dt, J = 24.4, 14.6 Hz, 2H), 6.13 (dd, J = 15.0, 9.9 Hz, 1H), 5.93 (dd, J = 24.5, 10.6 Hz, 1H), 5.52 (dd, J = 15.0, 8.8 Hz, 1H), 5.41 (d, J = 10.2 Hz, 1H), 5.35 (t, J = 4.7 Hz, 1H), 5.27 (d, J = 5.3 Hz, 1H), 5.16 (d, J = 4.9 Hz, 1H), 4.75 (s, 1H), 4.29- 4.07 (m, 3H), 3.93- 3.63 (m, 6H), 3.62- 3.47 (m, 3H), 3.46- 3.25 (m, 11H), 3.23- 3.01 (m, 3H), 2.79 (d, J = 4.9 Hz, 3H), 2.74 - 2.62 (m, 2H), 2.57 (dd, J = 16.5, 6.4 Hz, 1H), 2.34 (d, J = 12.7 Hz, 2H), 2.25- 2.19 (m, 1H), 2.08 (s, 1H), 1.93 (dd, J = 30.0, 22.7 Hz, 5H), 1.83- 1.65 (m, 7H), 1.55- 1.42 (m, 5H), 1.28 (s, 6H), 1.15- 0.83 (m, 18H), 0.72 (dd, J = 23.6, 12.0 Hz, 1H). I-74: ESI-MS (EI+, m/z): 1067.4 [M+Na]+. 1H NMR (400 MHz, CDCl3) d 6.50- 5.95 (m, 4H), 5.60- 4.96 (m, 5H), 4.59- 3.96 (m, 4H), 3.95- 3.66 (m, 6H), 3.64- 2.97 (m, 15H), 2.95- 2.65 (m, 6H), 2.59 (d, J = 11.0 Hz, 1H), 2.51- 1.95 (m, 5H), 1.78 (q, J = 6.8 Hz, 12H), 1.55- 1.29 (m, 11H), 1.15- 0.83 (m, 18H), 0.69 (dd, J = 23.6, 11.5 Hz, 1H).

Reference： [1]Patent: WO2019/241789,2019,A1 .Location in patent: Paragraph 00449-00454

37
2-[2-[tert-butyl(diphenyl)silyl]oxyethoxy]ethyl trifluoromethanesulfonate [ No CAS ]
[ 159351-69-6 ]
(35E,37E,39E,40E,50R,51S,52R,53R,55S,57S,59S,60S,61R,62R,71R)-60-[(1R)-2-[(1S,3R,4R)-4-[2-[2-[2-[tert-butyl(diphenyl)silyl]oxyethoxy]ethoxy]ethoxy]-3-methoxy-cyclohexyl]-1-methyl-ethyl]-61,71-dihydroxy-59,62-dimethoxy-50,51,52,53,63,64-hexamethyl-81,82-dioxa-73-azatricyclohexatriaconta-35,37,39⁽⁶³⁾,40⁽⁶⁴⁾-tetraene-65,66,67,68,69-pentone [ No CAS ]

Yield	Reaction Conditions	Operation in experiment
1.7 g	With N-ethyl-N,N-diisopropylamine; In toluene; at 60℃; for 18h;	A mixture of <strong>[159351-69-6]everolimus</strong> (2 g, 2.09 mmol), 2-[2-[tert- butyl(diphenyl)silyl]oxyethoxy]ethyl trifluoromethanesulfonate (9.95 g, 20.87 mmol) and N- ethyl-N-isopropyl-propan-2-amine (5.82 mL) in toluene (50 mL) was stirred at 60C for 18 h then poured into ice cold sat.NaHCO3 (60 mL). The reaction mixture was extracted with EtOAc (40 mL) and the organic layer washed with water (50 mL × 3) and brine (50 mL) then dried over anhydrous Na2SO4, filtered and concentrated. The residue was purified by silica gel chromatography (PE:EA = 5:1 to 3:1) to provide (35E,37E,39E,40E,50R,51S,52R,53R,55S,57S,59S,60S,61R,62R,71R)-60-[(1R)-2- [(1S,3R,4R)-4-[2-[2-[2-[tert-butyl(diphenyl)silyl]oxyethoxy]ethoxy]ethoxy]-3-methoxy- cyclohexyl]-1-methyl-ethyl]-61,71-dihydroxy-59,62-dimethoxy-50,51,52,53,63,64- hexamethyl-81,82-dioxa-73-azatricyclohexatriaconta-35,37,39(63),40(64)-tetraene- 65,66,67,68,69-pentone (1.7 g, 63% yield) as yellow solid. ESI-MS (EI+, m/z): 1307.5 [M+Na]+.

Reference： [1]Patent: WO2019/241789,2019,A1 .Location in patent: Paragraph 00467; 00470

38
[ 20356-03-0 ]
[ 159351-69-6 ]
(21E,23E,25E,26E,35R,36S,37R,38R,40S,42S,44S,45S,46R,47R,56R)-46,56-dihydroxy-45-[(1R)-2-[(1S,3R,4R)-4-(2-hydroxyethoxy)-3-methoxy-cyclohexyl]-1-methyl-ethyl]-47-methoxy-35,36,37,38,48,49-hexamethyl-44-[2-[2-(2,2,2-trifluoroethoxy)ethoxy]ethoxy]-67,68-dioxa-58-azatricyclohexatriaconta-21,23,25⁽⁴⁸⁾,26⁽⁴⁹⁾-tetraene-50,51,52,53,54-pentone [ No CAS ]
(21E,23E,25E,26E,35R,36S,37R,38R,40S,42S,44R,45S,46R,47R,56R)-46,56-dihydroxy-45-[(1R)-2-[(1S,3R,4R)-4-(2-hydroxyethoxy)-3-methoxy-cyclohexyl]-1-methyl-ethyl]-47-methoxy-35,36,37,38,48,49-hexamethyl-44-[2-[2-(2,2,2-trifluoroethoxy)ethoxy]ethoxy]-67,68-dioxa-58-azatricyclohexatriaconta-21,23,25⁽⁴⁸⁾,26⁽⁴⁹⁾-tetraene-50,51,52,53,54-pentone [ No CAS ]

Yield	Reaction Conditions	Operation in experiment
14.3 mg	With toluene-4-sulfonic acid; In tetrahydrofuran; at 0 - 25℃; for 6.5h;Inert atmosphere;	A solution of <strong>[159351-69-6]everolimus</strong> (0.5 g, 0.52 mmol) in THF (5 mL) was degassed, p- toluenesulfonic acid (0.45 g, 2.61 mmol) was added at 0 C followed by 2-[2-(2,2,2- trifluoroethoxy)ethoxy]ethanol (0.98 g, 5.22 mmol). The resulting mixture was stirred at 0C for 0.5 h under N2, then at 23 C for 6 h, poured into sat.NaHCO3 (40 mL) and extracted with EtOAc (30 mL). The organic layer was washed with water (30 mL × 2), brine (40 mL), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by reverse phase chromatography (C18, CH3CN:H2O from 0% to 70% yield) to provide (21E,23E,25E,26E,35R,36S,37R,38R,40S,42S,45S, 46R,47R,56R)-46,56-dihydroxy- 45-[(1R)-2-[(1S,3R,4R)-4-(2-hydroxyethoxy)-3-methoxy-cyclohexyl]-1-methyl-ethyl]-47- methoxy-35,36,37,38,48,49-hexamethyl-44-[2-[2-(2,2,2-trifluoroethoxy)ethoxy]ethoxy]- 67,68-dioxa-58-azatricyclohexatriaconta-21,23,25(48),26(49)-tetraene-50,51,52,53,54- pentone (I-86: 0.08 g, 14% yield) as a white solid. ESI-MS (EI+, m/z): 1136.5 [M+Na]+. 1H NMR (400MHz, CDCl3): d 6.44-5.88 (m, 4H), 5.73-5.06 (m, 4H), 4.52-4.32 (m, 1H), 4.22- 4.12 (m, 1H), 3.91-3.81 (m, 2H), 3.71-3.51 (m, 6H), 3.42-3.21 (m, 16H), 3.13-2.98 (m, 4H), 2.63-2.42 (m, 4H), 2.32-2.14 (m, 2H), 2.05-1.93 (m, 3H), 1.86-1.55 (m, 16H), 1.44-1.35 (m, 4H), 1.24-1.15 (m, 5H), 1.06-0.78 (m, 17H), 0.65-0.51 (m, 1H). 100 mg of (21E,23E,25E,26E,35R,36S,37R,38R,40S,42S,45S, 46R,47R,56R)- 46,56-dihydroxy-45-[(1R)-2-[(1S,3R,4R)-4-(2-hydroxyethoxy)-3-methoxy-cyclohexyl]-1- methyl-ethyl]-47-methoxy-35,36,37,38,48,49-hexamethyl-44-[2-[2-(2,2,2-trifluoroethoxy) ethoxy]ethoxy]-67,68-dioxa-58-azatricyclohexatriaconta-21,23,25(48),26(49)-tetraene- 50,51,52,53,54-pentone was purified via prep chiral HPLC which provided (21E,23E,25E,26E,35R,36S,37R,38R,40S,42S,44S,45S,46R,47R,56R)-46,56-dihydroxy-45- [(1R)-2-[(1S,3R,4R)-4-(2-hydroxyethoxy)-3-methoxy-cyclohexyl]-1-methyl-ethyl]-47- methoxy-35,36,37,38,48,49-hexamethyl-44-[2-[2-(2,2,2-trifluoroethoxy)ethoxy]ethoxy]- 67,68-dioxa-58-azatricyclohexatriaconta-21,23,25(48),26(49)-tetraene-50,51,52,53,54- pentone (I-85: 14.3 mg, 14.3% yield) as a white solid. Chiral separation method: Column : CHIRALPAK IC Column size : 5.0 cm I.D. × 25 cmL Solution concentration: 2.4 mg/ml Injection : 5 ml Mobile phase : Hexane/EtOH=70/30(V/V) Flow rate : 30 ml/min Wave length : UV 254 nm Temperature : 35 oC I-85: ESI-MS (EI+, m/z): 1136.4 [M+Na]+. 1H NMR (400 MHz, CDCl3) d 6.42- 6.06 (m, 3H), 5.92 (dd, J = 30.3, 10.3 Hz, 1H), 5.56- 5.06 (m, 5H), 4.74 (s, 1H), 4.18 (d, J = 5.7 Hz, 1H), 3.94- 3.83 (m, 2H), 3.82- 3.51 (m, 12H), 3.49- 3.25 (m, 11H), 3.22- 3.03 (m, 2H), 2.72 (dd, J = 16.6, 5.5 Hz, 2H), 2.57 (dd, J = 17.0, 6.5 Hz, 1H), 2.34 (d, J = 12.4 Hz, 2H), 2.25- 2.18 (m, 1H), 2.13- 1.85 (m, 5H), 1.69 (dd, J = 35.2, 8.9 Hz, 10H), 1.47 (dd, J = 20.5, 13.6 Hz, 5H), 1.26 (s, 7H), 1.15- 0.81 (m, 18H), 0.71 (dd, J = 23.9, 12.0 Hz, 1H).

Reference： [1]Patent: WO2019/241789,2019,A1 .Location in patent: Paragraph 00489; 00492-00495

39
2-[2-(trifluoromethoxy)ethoxy]ethanol [ No CAS ]
[ 159351-69-6 ]
(21E,23E,25E,26E,34R,35S,36R,37R,39S,41S,43R,44S,45R,46R,55R)-45,55-dihydroxy-44-[(1R)-2-[(1S,3R,4R)-4-(2-hydroxyethoxy)-3-methoxy-cyclohexyl]-1-methyl-ethyl]-46-methoxy-34,35,36,37,47,48-hexamethyl-43-[2-[2-(trifluoromethoxy)ethoxy]ethoxy]-66,67-dioxa-57-azatricyclohexatriaconta-21,23,25⁽⁴⁷⁾,26⁽⁴⁸⁾-tetraene-49,50,51,52,53-pentone [ No CAS ]
(21E,23E,25E,26E,34R,35S,36R,37R,39S,41S,43S,44S,45R,46R,55R)-45,55-dihydroxy-44-[(1R)-2-[(1S,3R,4R)-4-(2-hydroxyethoxy)-3-methoxy-cyclohexyl]-1-methyl-ethyl]-46-methoxy-34,35,36,37,47,48-hexamethyl-43-[2-[2-(trifluoromethoxy)ethoxy]ethoxy]-66,67-dioxa-57-azatricyclohexatriaconta-21,23,25⁽⁴⁷⁾,26⁽⁴⁸⁾-tetraene-49,50,51,52,53-pentone [ No CAS ]

Yield	Reaction Conditions	Operation in experiment
19 mg; 12 mg	With toluene-4-sulfonic acid; In tetrahydrofuran; at 0 - 25℃; for 6.5h;Inert atmosphere;	To a degassed solution of <strong>[159351-69-6]everolimus</strong> (0.5 g, 0.52 mmol) in THF (30 mL) was added p-toluenesulfonic acid (0.45 g, 2.61 mmol) at 0 C followed by 2-[2- (trifluoromethoxy)ethoxy]ethanol (0.91 g, 5.22 mmol). The resulting mixture was stirred at 0 C for 0.5 h under N2, then at 23C for 6 h. The reaction was poured into sat.NaHCO3 (40 mL) and extracted with EtOAc (30 mL). The organic layer was washed with water (30 mL × 2), brine (20 mL), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure at room temperature. The residue was purified by reverse phase chromatography (C18, CH3CN: H2O = 0% to 70% yield) to provide (21E,23E,25E,26E,34R,35S,36R,37R,39S, 41S,44S,45R,46R,55R)-45,55-dihydroxy-44-[(1R)-2-[(1S,3R,4R)-4-(2-hydroxyethoxy)-3- methoxy-cyclohexyl]-1-methyl-ethyl]-46-methoxy-34,35,36,37,47,48-hexamethyl-43-[2-[2- (trifluoromethoxy)ethoxy]ethoxy]-66,67-dioxa-57-azatricyclohexatriaconta- 21,23,25(47),26(48)-tetraene-49,50,51,52,53-pentone (I-88: 141 mg, 24% yield) as a white solid. ESI-MS (EI+, m/z): 1122.3 [M+Na]+. 1H NMR (400 MHz, CDCl3) d 6.39-5.88 (m, 4H), 5.73-5.05 (m, 5H), 4.52-3.83(m, 5H), 3.70-3.50 (m, 6H), 3.43-3.21 (m, 8H), 3.12-2.93 (m, 4H), 2.80-2.44 (m, 4H), 2.31-2.16 (m, 4H), 2.05-1.59 (m, 20H), 1.43-1.34 (m, 4H), 1.21-1.09 (m, 6H), 1.01-0.78 (m, 17H), 0.62-0.51 (m, 1H). 130 mg of (21E,23E,25E,26E,34R,35S,36R,37R,39S,41S,44S,45R,46R,55R)- 45,55-dihydroxy-44-[(1R)-2-[(1S,3R,4R)-4-(2-hydroxyethoxy)-3-methoxy-cyclohexyl]-1- methyl-ethyl]-46-methoxy-34,35,36,37,47,48-hexamethyl-43-[2-[2-(trifluoromethoxy) ethoxy]ethoxy]-66,67-dioxa-57-azatricyclohexatriaconta-21,23,25(47),26(48)-tetraene- 49,50,51,52,53-pentone was purified via prep chiral HPLC to provide (21E,23E,25E,26E,34R,35S,36R,37R,39S,41S,43R,44S,45R,46R,55R)-45,55-dihydroxy-44- [(1R)-2-[(1S,3R,4R)-4-(2-hydroxyethoxy)-3-methoxy-cyclohexyl]-1-methyl-ethyl]-46- methoxy-34,35,36,37,47,48-hexamethyl-43-[2-[2-(trifluoromethoxy)ethoxy]ethoxy]-66,67- dioxa-57-azatricyclohexatriaconta-21,23,25(47),26(48)-tetraene-49,50,51,52,53-pentone (I- 82: 19 mg, 14.6% yield) and (21E,23E,25E,26E,34R,35S,36R,37R,39S,41S,43S,44S,45R,46R,55R)-45,55-dihydroxy-44- [(1R)-2-[(1S,3R,4R)-4-(2-hydroxyethoxy)-3-methoxy-cyclohexyl]-1-methyl-ethyl]-46- methoxy-34,35,36,37,47,48-hexamethyl-43-[2-[2-(trifluoromethoxy)ethoxy]ethoxy]-66,67- dioxa-57-azatricyclohexatriaconta-21,23,25(47),26(48)-tetraene-49,50,51,52,53-pentone (I- 83: 12 mg, 9.2% yield), both as white solids. Chiral analysis method: I-82: ESI-MS (EI+, m/z): 1122.4 [M+Na]+. 1H NMR (500 MHz, CDCl3) d 6.43- 5.83 (m, 4H), 5.57- 5.13 (m, 5H), 4.31- 4.04 (m, 3H), 3.91- 3.53 (m, 11H), 3.49- 3.00 (m, 19H), 2.76- 2.52 (m, 3H), 2.25 (dd, J = 34.2, 26.6 Hz, 3H), 2.12- 1.96 (m, 5H), 1.75 (dd, J = 35.2, 24.7 Hz, 8H), 1.52- 1.34 (m, 8H), 1.15- 0.79 (m, 18H), 0.72 (d, J = 12.1 Hz, 1H). I-83: ESI-MS (EI+, m/z): 1122.4 [M+Na]+. 1H NMR (500 MHz, CDCl3) d 6.54- 6.41 (m, 1H), 6.18 (tdd, J = 29.7, 22.4, 12.7 Hz, 3H), 5.82- 5.68 (m, 1H), 5.46- 5.30 (m, 2H), 5.19 (dd, J = 25.5, 20.6 Hz, 2H), 4.62- 4.40 (m, 1H), 4.21 (d, J = 18.4 Hz, 1H), 3.94 (dd, J = 34.8, 4.5 Hz, 1H), 3.83- 3.62 (m, 4H), 3.59 (d, J = 3.3 Hz, 1H), 3.50- 2.95 (m, 13H), 2.62 (dt, J = 55.5, 38.6 Hz, 2H), 2.42- 2.17 (m, 3H), 2.16- 1.57 (m, 24H), 1.54- 1.27 (m, 10H), 1.12- 0.80 (m, 18H), 0.71- 0.62 (m, 1H).

Reference： [1]Patent: WO2019/241789,2019,A1 .Location in patent: Paragraph 00503; 00507-00511

40
2-[2-(1,1,2,2,2-pentafluoroethoxy)ethoxy]ethanol [ No CAS ]
[ 159351-69-6 ]
(21E,23E,25E,26E,34R,35S,36R,37R,39S,41S,43R,44S,45R,46R,55R)-45,55-dihydroxy-44-[(1R)-2-[(1S,3R,4R)-4-(2-hydroxyethoxy)-3-methoxy-cyclohexyl]-1-methyl-ethyl]-46-methoxy-34,35,36,37,47,48-hexamethyl-43-[2-[2-(1,1,2,2,2-pentafluoroethoxy)ethoxy]ethoxy]-67,68-dioxa-58-azatricyclohexatriaconta-21,23,25⁽⁴⁷⁾,26⁽⁴⁸⁾-tetraene-49,50,51,52,53-pentone [ No CAS ]
(21E,23E,25E,26E,34R,35S,36R,37R,39S,41S,43S,44S,45R,46R,55R)-45,55-dihydroxy-44-[(1R)-2-[(1S,3R,4R)-4-(2-hydroxyethoxy)-3-methoxy-cyclohexyl]-1-methyl-ethyl]-46-methoxy-34,35,36,37,47,48-hexamethyl-43-[2-[2-(1,1,2,2,2-pentafluoroethoxy)ethoxy]ethoxy]-67,68-dioxa-58-azatricyclohexatriaconta-21,23,25⁽⁴⁷⁾,26⁽⁴⁸⁾-tetraene-49,50,51,52,53-pentone [ No CAS ]

Yield	Reaction Conditions	Operation in experiment
5.1 mg; 7.2 mg	With toluene-4-sulfonic acid; In tetrahydrofuran; at 0 - 25℃; for 6.5h;	To a solution of <strong>[159351-69-6]everolimus</strong> (0.5 g, 0.52 mmol) in THF (10 mL) was added p- toluenesulfonic acid (0.45 g, 2.61 mmol) at 0 C, followed by 2-[2-(1,1,2,2,2- pentafluoroethoxy)ethoxy]ethanol (0.58 g, 2.61 mmol). The resulting mixture was stirred at 0 C for 0.5 h under N2, then at 22 C for 6 h, poured into sat. NaHCO3 (40 mL) and extracted with EtOAc (30 mL). The organic layer was washed with water (30 mL × 2), brine (20 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure at room temperature. The residue was purified by reverse phase chromatography (C18, CH3CN: H2O = 0% to 70% yield) to provide (21E,23E,25E,26E, 34R,35S,36R,37R,39S,41S,44S,45R,46R,55R)-45,55-dihydroxy-44-[(1R)-2-[(1S,3R,4R)-4- (2-hydroxyethoxy)-3-methoxy-cyclohexyl]-1-methyl-ethyl]-46-methoxy-34,35,36,37,47,48- hexamethyl-43-[2-[2-(1,1,2,2,2-pentafluoroethoxy)ethoxy]ethoxy]-67,68-dioxa-58- azatricyclohexatriaconta-21,23,25(47),26(48)-tetraene-49,50,51,52,53-pentone (I-87: 40 mg, 7% yield) as a white solid. ESI-MS (EI+, m/z): 1172.3 [M+Na]+. 1H NMR (400 MHz, CDCl3) d 6.45-5.95 (m, 4H), 5.79-5.13 (m, 5H), 4.59-4.17 (m, 3H), 3.98-3.59 (m, 8H), 3.50-3.28 (m, 10H), 3.20-3.00 (m, 5H), 2.89-2.49 (m, 4H), 2.37-2.22 (m, 4H), 2.10-1.61 (m, 20H), 1.51-1.23 (m, 8H), 1.06-0.85 (m, 16H), 0.71-0.63 (m, 1H). To a solution of <strong>[159351-69-6]everolimus</strong> (0.5 g, 0.52 mmol) in THF (10 mL) was added p- toluenesulfonic acid (0.45 g, 2.61 mmol) at 0 C, followed by 2-[2-(1,1,2,2,2- pentafluoroethoxy)ethoxy]ethanol (0.58 g, 2.61 mmol). The resulting mixture was stirred at 0 C for 0.5 h under N2, then at 22 C for 6 h, poured into sat. NaHCO3 (40 mL) and extracted with EtOAc (30 mL). The organic layer was washed with water (30 mL × 2), brine (20 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure at room temperature. The residue was purified by reverse phase chromatography (C18, CH3CN: H2O = 0% to 70% yield) to provide (21E,23E,25E,26E, 34R,35S,36R,37R,39S,41S,44S,45R,46R,55R)-45,55-dihydroxy-44-[(1R)-2-[(1S,3R,4R)-4- (2-hydroxyethoxy)-3-methoxy-cyclohexyl]-1-methyl-ethyl]-46-methoxy-34,35,36,37,47,48- hexamethyl-43-[2-[2-(1,1,2,2,2-pentafluoroethoxy)ethoxy]ethoxy]-67,68-dioxa-58- azatricyclohexatriaconta-21,23,25(47),26(48)-tetraene-49,50,51,52,53-pentone (I-87: 40 mg, 7% yield) as a white solid. ESI-MS (EI+, m/z): 1172.3 [M+Na]+. 1H NMR (400 MHz, CDCl3) d 6.45-5.95 (m, 4H), 5.79-5.13 (m, 5H), 4.59-4.17 (m, 3H), 3.98-3.59 (m, 8H), 3.50-3.28 (m, 10H), 3.20-3.00 (m, 5H), 2.89-2.49 (m, 4H), 2.37-2.22 (m, 4H), 2.10-1.61 (m, 20H), 1.51-1.23 (m, 8H), 1.06-0.85 (m, 16H), 0.71-0.63 (m, 1H). Chiral separation method: Column : CHIRALPAK IC Column size : 5.0 cm I.D. × 25 cm L I-80: ESI-MS (EI+, m/z): 1150.3 [M+H]+. 1H NMR (400 MHz, CDCl3) d 6.42- 5.80 (m, 4H), 5.58- 5.05 (m, 5H), 4.74 (s, 1H), 4.24- 4.08 (m, 3H), 3.92- 3.51 (m, 10H), 3.48- 3.24 (m, 12H), 3.13 (ddd, J = 24.2, 17.1, 11.3 Hz, 3H), 2.64 (ddd, J = 23.4, 16.8, 6.0 Hz, 3H), 2.42- 2.15 (m, 3H), 2.14- 1.88 (m, 6H), 1.84- 1.64 (m, 14H), 1.54- 1.39 (m, 5H), 1.17- 0.81 (m, 18H), 0.71 (dd, J = 23.8, 12.1 Hz, 1H). I-79: ESI-MS (EI+, m/z): 1172.3 [M+Na]+. 1H NMR (400 MHz, CDCl3) d 6.42- 5.90 (m, 4H), 5.54- 5.03 (m, 6H), 4.31- 3.96 (m, 5H), 3.88- 3.01 (m, 22H), 2.94- 2.37 (m, 5H), 2.39- 1.94 (m, 7H), 1.68 (dd, J = 28.2, 19.2 Hz, 8H), 1.52- 1.31 (m, 6H), 1.14- 1.00 (m, 6H), 0.97- 0.62 (m, 19H).

Reference： [1]Patent: WO2019/241789,2019,A1 .Location in patent: Paragraph 00512; 00515-00519

41
2-[tert-butyl(diphenyl)silyl]oxy}ethyl trifluoromethansulphonate [ No CAS ]
[ 159351-69-6 ]
(35E,37E,39E,40E,48R,49S,50R,51R,53S,55S,57S,58S,59R,60R,69R)-58-[(1R)-2-[(1S,3R,4R)-4-[2-[2-[tert-butyl(diphenyl)silyl]oxyethoxy]ethoxy]-3-methoxy-cyclohexyl]-1-methyl-ethyl]-59,69-dihydroxy-57,60-dimethoxy-48,49,50,51,61,62-hexamethyl-79,80-dioxa-71-azatricyclohexatriaconta-35,37,39⁽⁶¹⁾,40⁽⁶²⁾-tetraene-63,64,65,66,67-pentone [ No CAS ]

Yield	Reaction Conditions	Operation in experiment
59%	With N-ethyl-N,N-diisopropylamine; In toluene; at 45℃; for 18h;	A solution of <strong>[159351-69-6]everolimus</strong> (1.5 g, 1.57 mmol), 2-[tert-butyl(diphenyl)silyl]oxyethyl trifluoromethanesulfonate and DIPEA (3.27 mL, 18.78 mmol) in toluene (20 mL) was stirred at 45 C for 18 h. The mixture was then poured into ice cold sat. NaHCO3 (50 mL), washed with ice-water twice (60 mL), brine (50 mL), dried over anhydrous Na2SO4, filtered, and concentrated. The mixture was purified via silica gel chromatography (PE:EtOAc = 5:1 to 2:1, then PE: acetone = 4:1) to obtain (35E,37E,39E,40E,48R,49S,50R,51R,53S,55S,57S,58S,59R,60R,69R)-58-[(1R)-2- [(1S,3R,4R)-4-[2-[2-[tert-butyl(diphenyl)silyl]oxyethoxy]ethoxy]-3-methoxy-cyclohexyl]-1- methyl-ethyl]-59,69-dihydroxy-57,60-dimethoxy-48,49,50,51,61,62-hexamethyl-79,80- dioxa-71-azatricyclohexatriaconta-35,37,39(61),40(62)-tetraene-63,64,65,66,67-pentone (1.15 g, 59% yield) as a brown solid. ESI-MS (EI+, m/z): 1263.4 [M+Na]+. 1H NMR (400 MHz, CDCl3) d 7.62-7.60 (m, 4H), 7.35-7.28 (m, 6H), 6.35-5.80 (m, 4H), 5.51-5.09 (m, 4H), 4.75 (s, 1H), 4.37-4.02 (m, 2H), 3.87-3.49 (m, 11H), 3.37-3.26 (m, 8H), 3.10-2.96 (m, 5H), 2.76-2.48 (m, 3H), 2.28-2.21 (m, 2H), 1.98-1.90 (m, 3H), 1.67-1.39 (m, 18H), 1.26-1.08 (m, 7H), 1.04-0.76 (m, 26H), 0.64 (q, J = 11.2Hz, 1H).

Reference： [1]Patent: WO2019/241789,2019,A1 .Location in patent: Paragraph 00520; 00522

42
[ 112-60-7 ]
[ 159351-69-6 ]
(21E,23E,25E,26E,38R,39S,40R,41R,43S,45S,48S,49R,50R,59R)-49,59-dihydroxy-47-[2-[2-[2-(2-hydroxyethoxy)ethoxy]ethoxy]ethoxy]-48-[(1R)-2-[(1S,3R,4R)-4-(2-hydroxyethoxy)-3-methoxy-cyclohexyl]-1-methyl-ethyl]-50-methoxy-38,39,40,41,51,52-hexamethyl-70,71-dioxa-60-azatricyclohexatriaconta-21,23,25⁽⁵¹⁾,26⁽⁵²⁾-tetraene-53,54,55,56,57-pentone [ No CAS ]

Yield	Reaction Conditions	Operation in experiment
17%	With toluene-4-sulfonic acid; In tetrahydrofuran; water; at 20℃; for 2h;	A mixture of <strong>[159351-69-6]everolimus</strong> (0.5 g, 0.52 mmol), 2-[2-[2-(2- hydroxyethoxy)ethoxy]ethoxy]ethanol (2.03 g, 10.44 mmol) and p-toluenesulfonic acid hydrate (0.54 g, 3.13 mmol) in THF (6 mL) was stirred at 20 C for 2 h. The mixture was poured into ice cold saturated NaHCO3 (30 mL) and extracted with EtOAc (50 mL*3). The combined organic layers were washed with water, brine, then concentrated in vacuo. The residue was purified by reversed phase chromatography (CH3CN/pure water = 1:1) to afford (21E,23E,25E,26E,38R,39S,40R,41R,42S,45S,47S,49R,50R,59R)-49,59-dihydroxy-46-[2-[2- [2-(2-hydroxyethoxy)ethoxy]ethoxy]ethoxy]-47-[(1R)-2-[(1S,2R,3R)-3-(2-hydroxyethoxy)- 2-methoxy-cyclohexyl]-1-methyl-ethyl]-50-methoxy-38,39,40,41,51,52-hexamethyl-70,71- dioxa-60-azatricyclohexatriaconta-21,23,25(51),26(52)-tetraene-53,54,55,56,57-pentone (I- 22: 0.1 g, 17% yield) as a white solid. ESI-MS (EI+, m/z): 1043.4 [M+Na]+. 1H NMR (500 MHz, CDCl3) d 6.71- 5.78 (m, 4H), 5.77- 5.01 (m, 4H), 4.65- 3.88 (m, 3H), 3.87- 3.50 (m, 20H), 3.50- 3.00 (m, 14H), 2.77- 1.95 (m, 11H), 1.89- 1.53 (m, 13H), 1.53- 0.79 (m, 27H), 0.76- 0.62 (m, 1H).

Reference： [1]Patent: WO2019/241789,2019,A1 .Location in patent: Paragraph 00356-00357

43
[ 112-60-7 ]
[ 159351-69-6 ]
(21E,23E,25E,26E,38R,39S,40R,41R,43S,45S,47R,48S,49R,50R,59R)-49,59-dihydroxy-47-[2-[2-[2-(2-hydroxyethoxy)ethoxy]ethoxy]ethoxy]-48-[(1R)-2-[(1S,3R,4R)-4-(2-hydroxyethoxy)-3-methoxy-cyclohexyl]-1-methyl-ethyl]-50-methoxy-38,39,40,41,51,52-hexamethyl-70,71-dioxa-60-azatricyclohexatriaconta-21,23,25⁽⁵¹⁾,26⁽⁵²⁾-tetraene-53,54,55,56,57-pentone [ No CAS ]
(21E,23E,25E,26E,38R,39S,40R,41R,43S,45S,47S,48S,49R,50R,59R)-49,59-dihydroxy-47-[2-[2-[2-(2-hydroxyethoxy)ethoxy]ethoxy]ethoxy]-48-[(1R)-2-[(1S,3R,4R)-4-(2-hydroxyethoxy)-3-methoxy-cyclohexyl]-1-methyl-ethyl]-50-methoxy-38,39,40,41,51,52-hexamethyl-70,71-dioxa-60-azatricyclohexatriaconta-21,23,25⁽⁵¹⁾,26⁽⁵²⁾-tetraene-53,54,55,56,57-pentone [ No CAS ]

Yield	Reaction Conditions	Operation in experiment
300 mg; 563 mg	With toluene-4-sulfonic acid; In tetrahydrofuran; at 0 - 25℃; for 3h;	A mixture of <strong>[159351-69-6]everolimus</strong> (1 g, 1.04 mmol), 2-[2-[2-(2- hydroxyethoxy)ethoxy]ethoxy]ethanol (4.05 g, 20.87 mmol) and p-toluenesulfonic acid (0.898 g, 5.22 mmol) in THF (20 mL) was stirred at 20 C for 2 h. The mixture was then poured into cold sat.NaHCO3 (30 mL), extracted with EtOAc (50 mL × 3) and the combined organic layers washed with water and brine then dried over anhydrous Na2SO4, filtered and concentrated. The resulting residue was purified by reversed phase chromatography (C18, CH3CN: H2O= 6:4) to afford (21E,23E,25E,26E,38R,39S,40R,41R,43S,45S,48S,49R,50R,59R)-49,59-dihydroxy- 47-[2-[2-[2-(2-hydroxyethoxy)ethoxy]ethoxy]ethoxy]-48-[(1R)-2-[(1S,3R,4R)-4-(2- hydroxyethoxy)-3-methoxy-cyclohexyl]-1-methyl-ethyl]-50-methoxy-38,39,40,41,51,52- hexamethyl-70,71-dioxa-60-azatricyclohexatriaconta-21,23,25(51),26(52)-tetraene- 53,54,55,56,57-pentone (0.25 g, 21% yield) as white solid. ESI-MS (EI+, m/z):1142.4 [M+Na]+. 1H NMR (400 MHz, CDCl3) d 6.41- 5.85 (m, 4H), 5.78 (s, 1H), 5.60- 4.98 (m, 4H), 4.22 (t, J = 27.1 Hz, 1H), 3.97 (dd, J = 17.7, 6.4 Hz, 1H), 3.87- 3.54 (m, 21H), 3.51- 3.03 (m, 15H), 2.72- 2.45 (m, 3H), 2.28 (s, 6H), 2.16- 1.96 (m, 4H), 1.89- 1.56 (m, 10H), 1.51- 1.17 (m, 8H), 1.17- 0.81 (m, 18H), 0.78- 0.62 (m, 1H). 1.5 g of (21E,23E,25E,26E,38R,39S,40R,41R,43S,45S,48S,49R,50R,59R)-49,59- dihydroxy-47-[2-[2-[2-(2-hydroxyethoxy)ethoxy]ethoxy]ethoxy]-48-[(1R)-2-[(1S,3R,4R)-4- (2-hydroxyethoxy)-3-methoxy-cyclohexyl]-1-methyl-ethyl]-50-methoxy-38,39,40,41,51,52- hexamethyl-70,71-dioxa-60-azatricyclohexatriaconta-21,23,25(51),26(52)-tetraene- 53,54,55,56,57-pentone was purified via prep chiral HPLC and the resulting epimers purified via silica gel chromatography (hexane:DCM:EtOAc:MeOH = 8:8:3:1.2) to obtain (21E,23E,25E,26E,38R,39S,40R,41R,43S,45S,47R,48S,49R,50R,59R)-49,59-dihydroxy-47- [2-[2-[2-(2-hydroxyethoxy)ethoxy]ethoxy]ethoxy]-48-[(1R)-2-[(1S,3R,4R)-4-(2- hydroxyethoxy)-3-methoxy-cyclohexyl]-1-methyl-ethyl]-50-methoxy-38,39,40,41,51,52- hexamethyl-70,71-dioxa-60-azatricyclohexatriaconta-21,23,25(51),26(52)-tetraene- 53,54,55,56,57-pentone (I-114: 300 mg, 20% yield) and (21E,23E,25E,26E,38R,39S,40R,41R,43S,45S,47S,48S,49R,50R,59R)-49,59-dihydroxy-47- [2-[2-[2-(2-hydroxyethoxy)ethoxy]ethoxy]ethoxy]-48-[(1R)-2-[(1S,3R,4R)-4-(2- hydroxyethoxy)-3-methoxy-cyclohexyl]-1-methyl-ethyl]-50-methoxy-38,39,40,41,51,52- hexamethyl-70,71-dioxa-60-azatricyclohexatriaconta-21,23,25(51),26(52)-tetraene- 53,54,55,56,57-pentone (I-115: 563 mg, 38% yield) as a white solids. Chiral analysis conditions: I-114: ESI-MS (EI+, m/z):1142.5 [M+Na]+. 1H NMR (500 MHz, CDCl3) d 6.42- 5.91 (m, 4H), 5.60- 5.08 (m, 4H), 4.19 (dd, J = 43.8, 32.6 Hz, 1H), 3.96 (dd, J = 27.1, 6.3 Hz, 1H), 3.85- 3.54 (m, 21H), 3.53- 3.01 (m, 12H), 2.93- 2.80 (m, 1H), 2.75- 2.45 (m, 3H), 2.30 (d, J = 12.1 Hz, 1H), 2.03 (dd, J = 37.0, 32.8 Hz, 13H), 1.84- 1.69 (m, 12H), 1.50 - 1.18 (m, 5H), 1.16- 0.82 (m, 18H), 0.71 (dt, J = 23.8, 12.1 Hz, 1H). I-115: ESI-MS (EI+, m/z):1142.4 [M+Na]+. 1H NMR (500 MHz, CDCl3) d 6.36 (dd, J = 14.7, 10.9 Hz, 1H), 6.23 (dd, J = 14.7, 10.6 Hz, 1H), 6.10 (dd, J = 15.0, 10.5 Hz, 1H), 5.92 (dd, J = 45.2, 10.5 Hz, 1H), 5.61 (s, 1H), 5.43 (dd, J = 15.6, 9.6 Hz, 2H), 5.21 (d, J = 5.4 Hz, 1H), 5.10 (dd, J = 9.9, 5.9 Hz, 1H), 4.20 (d, J = 4.5 Hz, 1H), 4.10- 3.95 (m, 1H), 3.84 (d, J = 5.0 Hz, 1H), 3.80- 3.49 (m, 21H), 3.48- 3.13 (m, 12H), 3.11- 3.01 (m, 1H), 2.73- 2.50 (m, 3H), 2.34- 2.19 (m, 2H), 2.01 (ddd, J = 62.0, 34.6, 28.2 Hz, 12H), 1.80- 1.54 (m, 10H), 1.51- 1.37 (m, 5H), 1.35- 1.12 (m, 6H), 1.05 (dd, J = 6.4, 5.0 Hz, 6H), 0.97 (d, J = 6.5 Hz, 3H), 0.93 (d, J = 6.6 Hz, 3H), 0.91- 0.82 (m, 6H), 0.74- 0.67 (m, 1H).

Reference： [1]Patent: WO2019/241789,2019,A1 .Location in patent: Paragraph 00391-00396

44
[ 18190-25-5 ]
[ 159351-69-6 ]
4-[[(23E,25E,27E,28E,35R,36S,37R,38R,40S,42S,44R,45S,46R,47R,57R)-46,57-dihydroxy-45-[(1R)-2-[(1S,3R,4R)-4-(2-hydroxyethoxy)-3-methoxy-cyclohexyl]-1-methyl-ethyl]-47-methoxy-35,36,37,38,48,49-hexamethyl-50,51,52,53,54-pentaoxo-69,70-dioxa-58-azatricyclohexatriaconta-23,25,27⁽⁴⁸⁾,28⁽⁴⁹⁾-tetraen-44-yl]oxy]-N,N-dimethyl-butanamide [ No CAS ]
4-[[(23E,25E,27E,28E,35R,36S,37R,38R,40S,42S,44S,45S,46R,47R,57R)-46,57-dihydroxy-45-[(1R)-2-[(1S,3R,4R)-4-(2-hydroxyethoxy)-3-methoxy-cyclohexyl]-1-methyl-ethyl]-47-methoxy-35,36,37,38,48,49-hexamethyl-50,51,52,53,54-pentaoxo-69,70-dioxa-58-azatricyclohexatriaconta-23,25,27⁽⁴⁸⁾,28⁽⁴⁹⁾-tetraen-44-yl]oxy]-N,N-dimethyl-butanamide [ No CAS ]

Yield	Reaction Conditions	Operation in experiment
24.2 mg; 34.3 mg	With toluene-4-sulfonic acid; In tetrahydrofuran; at 0 - 35℃; for 18h;Inert atmosphere;	To a solution of <strong>[159351-69-6]everolimus</strong> (0.3 g, 0.313 mmol) in THF (9 mL) at 0 C under N2was added p-toluenesulfonic acid (0.27 g, 1.57 mmol) and 4-hydroxy-N,N-dimethyl-butanamide (0.82 g, 6.26 mmol). The mixture was warmed to 35 C and stirred for 18 h then poured into ice cold NaHCO3 and extracted with EtOAc (20 mL × 3). The combined organic layers were washed with water (30 mL), brine (30 mL), dried over anhydrous Na2SO4, filtered, and concentrated. The residue was purified by reverse phase chromatography (C18, CH3CN:H2O= 55:45) to provide 4-[[(23E,25E,27E,28E,35R,36S,37R,38R,40S,42S,45S,46R,47R,57R)- 46,57-dihydroxy-45-[(1R)-2-[(1S,3R,4R)-4-(2-hydroxyethoxy)-3-methoxy-cyclohexyl]-1- methyl-ethyl]-47-methoxy-35,36,37,38,48,49-hexamethyl-50,51,52,53,54-pentaoxo-69,70- dioxa-58-azatricyclohexatriaconta-23,25,27(48),28(49)-tetraen-44-yl]oxy]-N,N-dimethyl- butanamide (I-69: 0.05 g, 15% yield) as a white solid. ESI-MS (EI+, m/z): 1079.4 [M+Na]+. 1H NMR (400 MHz, CDCl3) d 6.52- 5.78 (m, 4H), 5.67- 5.05 (m, 4H), 4.75 (s, 1H), 4.48- 4.09 (m, 2H), 4.04- 3.51 (m, 7H), 3.52- 3.12 (m, 12H), 3.12- 2.88 (m, 7H), 2.82- 2.27 (m, 6H), 2.22- 1.53 (m, 23H), 1.54- 1.13 (m, 10H), 1.12- 0.80 (m, 15H), 0.79- 0.61 (m, 1H). 120 mg of 4-[[(23E,25E,27E,28E,35R,36S,37R,38R,40S,42S,45S,46R,47R,57R)- 46,57-dihydroxy-45-[(1R)-2-[(1S,3R,4R)-4-(2-hydroxyethoxy)-3-methoxy-cyclohexyl]-1- methyl-ethyl]-47-methoxy-35,36,37,38,48,49-hexamethyl-50,51,52,53,54-pentaoxo-69,70- dioxa-58-azatricyclohexatriaconta-23,25,27(48),28(49)-tetraen-44-yl]oxy]-N,N-dimethyl- butanamide was purified via prep chiral HPLC to provide 4- [[(23E,25E,27E,28E,35R,36S,37R,38R,40S,42S,44S,45S,46R,47R,57R)-46,57-dihydroxy- 45-[(1R)-2-[(1S,3R,4R)-4-(2-hydroxyethoxy)-3-methoxy-cyclohexyl]-1-methyl-ethyl]-47- methoxy-35,36,37,38,48,49-hexamethyl-50,51,52,53,54-pentaoxo-69,70-dioxa-58- azatricyclohexatriaconta-23,25,27(48),28(49)-tetraen-44-yl]oxy]-N,N-dimethyl-butanamide (I-62: 34.3 mg, 29% yield) and 4- [[(23E,25E,27E,28E,35R,36S,37R,38R,40S,42S,44R,45S,46R,47R,57R)-46,57-dihydroxy- 45-[(1R)-2-[(1S,3R,4R)-4-(2-hydroxyethoxy)-3-methoxy-cyclohexyl]-1-methyl-ethyl]-47- methoxy-35,36,37,38,48,49-hexamethyl-50,51,52,53,54-pentaoxo-69,70-dioxa-58- azatricyclohexatriaconta-23,25,27(48),28(49)-tetraen-44-yl]oxy]-N,N-dimethyl-butanamide (I-61: 24.2 mg, 20% yield), both as white solids. Chiral separation method: I-62: ESI-MS (EI+, m/z): 1079.4 [M+Na]+. 1H NMR (400 MHz, CDCl3) d 6.30 (tt, J = 34.4, 17.0 Hz, 2H), 6.13 (dd, J = 14.9, 10.0 Hz, 1H), 5.91 (dd, J = 28.8, 10.6 Hz, 1H), 5.51 (dd, J = 15.0, 8.9 Hz, 1H), 5.45- 5.37 (m, 1H), 5.27 (d, J = 5.4 Hz, 1H), 5.17 (d, J = 4.4 Hz, 1H), 4.75 (s, 1H), 4.19 (d, J = 4.9 Hz, 1H), 3.88 (s, 1H), 3.82- 3.64 (m, 5H), 3.62- 3.52 (m, 2H), 3.47- 3.25 (m, 11H), 3.24- 3.14 (m, 2H), 3.10 (d, J = 7.0 Hz, 1H), 3.01 (s, 3H), 2.94 (s, 3H), 2.71 (dd, J = 16.7, 5.7 Hz, 2H), 2.55 (dd, J = 16.8, 6.6 Hz, 1H), 2.44- 2.25 (m, 4H), 2.14- 1.63 (m, 17H), 1.33 (ddd, J = 40.8, 27.4, 12.3 Hz, 11H), 1.14- 0.83 (m, 18H), 0.71 (dd, J = 23.8, 11.9 Hz, 1H). I-61: ESI-MS (EI+, m/z): 1079.3 [M+Na]+. 1H NMR (400 MHz, CDCl3) d 6.43- 5.88 (m, 4H), 5.65- 5.08 (m, 5H), 4.33- 4.08 (m, 2H), 3.94- 3.52 (m, 6H), 3.49- 3.31 (m, 8H), 3.30- 3.12 (m, 8H), 3.09- 2.81 (m, 8H), 2.75- 2.26 (m, 6H), 2.10 (d, J = 63.9 Hz, 3H), 1.88- 1.65 (m, 14H), 1.35 (dt, J = 49.7, 11.3 Hz, 9H), 1.18- 0.81 (m, 18H), 0.77- 0.60 (m, 1H).

Reference： [1]Patent: WO2019/241789,2019,A1 .Location in patent: Paragraph 00542; 00544-00548

45
[ 37941-69-8 ]
[ 159351-69-6 ]
4-[[(22E,24E,26E,27E,34R,35S,36R,37R,39S,41S,43R,44S,45R,46R,56R)-45,56-dihydroxy-44-[(1R)-2-[(1S,3R,4R)-4-(2-hydroxyethoxy)-3-methoxy-cyclohexyl]-1-methyl-ethyl]-46-methoxy-34,35,36,37,47,48-hexamethyl-49,50,51,52,53-pentaoxo-68,69-dioxa-58-azatricyclohexatriaconta-22,24,26⁽⁴⁷⁾,27⁽⁴⁸⁾-tetraen-43-yl]oxy]-N-methyl-butanamide [ No CAS ]
4-[[(22E,24E,26E,27E,34R,35S,36R,37R,39S,41S,43S,44S,45R,46R,56R)-45,56-dihydroxy-44-[(1R)-2-[(1S,3R,4R)-4-(2-hydroxyethoxy)-3-methoxy-cyclohexyl]-1-methyl-ethyl]-46-methoxy-34,35,36,37,47,48-hexamethyl-49,50,51,52,53-pentaoxo-68,69-dioxa-58-azatricyclohexatriaconta-22,24,26⁽⁴⁷⁾,27⁽⁴⁸⁾-tetraen-43-yl]oxy]-N-methyl-butanamide [ No CAS ]

Yield	Reaction Conditions	Operation in experiment
36 mg; 25 mg	With toluene-4-sulfonic acid; In tetrahydrofuran; at 0 - 35℃; for 18h;Inert atmosphere;	A solution of <strong>[159351-69-6]everolimus</strong> (1 g, 1.04 mmol) and 4-hydroxy-N-methyl-butanamide (2.45 g, 20.87 mmol) in THF (30 mL) was cooled to 0 C under N2 and p-toluenesulfonic acid (0.9 g, 5.22 mmol) added. The reaction was warmed to 35 C and stirred for 18 h then poured into sat. NaHCO3 (150 mL) and extracted with EtOAc (100 mL× 3). The combined organic layers were washed with water (80 mL), brine (80 mL), dried over anhydrous Na2SO4, filtered and concentrated. The residue was purified by reverse phase chromatography (C18, 80 g, CH3CN:H2O = 37:33) to provide 4- [[(22E,24E,26E,27E,34R,35S,36R,37R,39S,41S,44S,45R,46R,56R)-45,56-dihydroxy-44- [(1R)-2-[(1S,3R,4R)-4-(2-hydroxyethoxy)-3-methoxy-cyclohexyl]-1-methyl-ethyl]-46- methoxy-34,35,36,37,47,48-hexamethyl-49,50,51,52,53-pentaoxo-68,69-dioxa-58- azatricyclohexatriaconta-22,24,26(47),27(48)-tetraen-43-yl]oxy]-N-methyl-butanamide (I-68: 0.14 g, 13% yield) as a white solid. ESI-MS (EI+, m/z): 1065.3 [M+Na]+. 1H NMR (400 MHz, CDCl3) d 6.50- 5.85 (m, 4H), 5.78- 4.96 (m, 5H), 4.78 (s, 1H), 4.33- 4.03 (m, 2H), 3.98- 3.64 (m, 5H), 3.63- 3.49 (m, 2H), 3.49- 2.90 (m, 13H), 2.90- 2.48 (m, 6H), 2.41- 1.94 (m, 7H), 1.93- 1.54 (m, 18H), 1.53- 1.11 (m, 10H), 1.11- 0.80 (m, 16H), 0.78- 0.54 (m, 1H). 130mg of 4-[[(22E,24E,26E,27E,34R,35S,36R,37R,39S,41S,44S,45R,46R,56R)- 45,56-dihydroxy-44-[(1R)-2-[(1S,3R,4R)-4-(2-hydroxyethoxy)-3-methoxy-cyclohexyl]-1- methyl-ethyl]-46-methoxy-34,35,36,37,47,48-hexamethyl-49,50,51,52,53-pentaoxo-68,69- dioxa-58-azatricyclohexatriaconta-22,24,26(47),27(48)-tetraen-43-yl]oxy]-N-methyl- butanamide was purified via prep chiral HPLC and the resulting epimers purified via silica gel chromatography (hexane:DCM:EtOAc:MeOH = 3:3:1:0.8) to provide 4- [[(22E,24E,26E,27E,34R,35S,36R,37R,39S,41S,43S,44S,45R,46R,56R)-45,56-dihydroxy- 44-[(1R)-2-[(1S,3R,4R)-4-(2-hydroxyethoxy)-3-methoxy-cyclohexyl]-1-methyl-ethyl]-46- methoxy-34,35,36,37,47,48-hexamethyl-49,50,51,52,53-pentaoxo-68,69-dioxa-58- azatricyclohexatriaconta-22,24,26(47),27(48)-tetraen-43-yl]oxy]-N-methyl-butanamide (I-60: 25 mg, 19% yield) and 4- [[(22E,24E,26E,27E,34R,35S,36R,37R,39S,41S,43R,44S,45R,46R,56R)-45,56-dihydroxy- 44-[(1R)-2-[(1S,3R,4R)-4-(2-hydroxyethoxy)-3-methoxy-cyclohexyl]-1-methyl-ethyl]-46- methoxy-34,35,36,37,47,48-hexamethyl-49,50,51,52,53-pentaoxo-68,69-dioxa-58- azatricyclohexatriaconta-22,24,26(47),27(48)-tetraen-43-yl]oxy]-N-methyl-butanamide (I-59: 36 mg, 27% yield), both as white solids. Chiral analysis method: I HPLC equipment: Shimadzu LC-20AT CP-HPLC-06 I-60: ESI-MS (EI+, m/z): 1065.4 [M+Na]+. 1H NMR (400 MHz, CDCl3) d 6.32 (ddd, J = 31.4, 14.8, 10.2 Hz, 2H), 6.13 (dd, J = 15.1, 9.9 Hz, 1H), 6.00- 5.85 (m, 1H), 5.69 (s, 1H), 5.51 (dd, J = 14.9, 9.0 Hz, 1H), 5.41 (d, J = 9.9 Hz, 1H), 5.27 (d, J = 5.7 Hz, 1H), 5.13 (dt, J = 48.5, 24.3 Hz, 1H), 4.77 (s, 1H), 4.18 (d, J = 5.7 Hz, 1H), 3.92- 3.63 (m, 6H), 3.61- 3.50 (m, 2H), 3.46- 3.25 (m, 10H), 3.22- 3.00 (m, 3H), 2.79 (dd, J = 4.8, 2.2 Hz, 3H), 2.72 (dd, J = 16.9, 5.5 Hz, 2H), 2.55 (dd, J = 16.8, 6.5 Hz, 1H), 2.38- 2.14 (m, 4H), 2.12- 1.91 (m, 4H), 1.89- 1.62 (m, 15H), 1.52- 1.11 (m, 13H), 1.10- 1.01 (m, 6H), 1.00- 0.81 (m, 9H), 0.71 (dd, J = 23.6, 12.1 Hz, 1H). I-59: ESI-MS (EI+, m/z): 1065.4 [M+Na]+. 1H NMR (400 MHz, CDCl3) d 6.37- 5.81 (m, 4H), 5.66 (d, J = 15.6 Hz, 1H), 5.60- 4.89 (m, 4H), 4.16 (ddd, J = 31.9, 15.6, 5.8 Hz, 2H), 3.93- 3.42 (m, 8H), 3.38- 2.95 (m, 12H), 2.89- 2.39 (m, 7H), 2.37- 1.89 (m, 9H), 1.83 - 1.60 (m, 13H), 1.45- 1.05 (m, 11H), 1.03- 0.73 (m, 18H), 0.62 (dd, J = 23.9, 12.1 Hz, 1H).

Reference： [1]Patent: WO2019/241789,2019,A1 .Location in patent: Paragraph 00549; 00551-00555

46
[ 159351-69-6 ]
(22E,24E,26E,27E,35R,36S,37R,38R,40S,42S,44S,45S,46R,47R,56R)-46,56-dihydroxy-45-[(1R)-2-[(1S,3R,4R)-4-[2-[2-(2-hydroxyethoxy)ethoxy]ethoxy]-3-methoxy-cyclohexyl]-1-methyl-ethyl]-44,47-dimethoxy-35,36,37,38,48,49-hexamethyl-66,67-dioxa-57-azatricyclohexatriaconta-22,24,26⁽⁴⁸⁾,27⁽⁴⁹⁾-tetraene-50,51,52,53,54-pentone [ No CAS ]

Reference： [1]Patent: WO2019/241789,2019,A1

47
[ 159351-69-6 ]
(22E,24E,26E,27E,33R,34S,35R,36R,38S,40S,42S,43S,44R,45R,54R)-44,54-dihydroxy-43-[(1R)-2-[(1S,3R,4R)-4-[2-(2-hydroxyethoxy)ethoxy]-3-methoxy-cyclohexyl]-1-methyl-ethyl]-42,45-dimethoxy-33,34,35,36,46,47-hexamethyl-64,65-dioxa-55-azatricyclohexatriaconta-22,24,26⁽⁴⁶⁾,27⁽⁴⁷⁾-tetraene-48,49,50,51,52-pentone [ No CAS ]

Reference： [1]Patent: WO2019/241789,2019,A1

48
[ 159351-69-6 ]
(21E,23E,25E,26E,36R,37S,38R,39R,41S,43S,45S,46S,47R,48R,57R)-47,57-dihydroxy-45-[2-(2-hydroxyethoxy)ethoxy]-46-[(1R)-2-[(1S,3R,4R)-4-[2-(2-hydroxyethoxy)ethoxy]-3-methoxy-cyclohexyl]-1-methyl-ethyl]-48-methoxy-36,37,38,39,49,50-hexamethyl-68,69-dioxa-58-azatricyclohexatriaconta-21,23,25⁽⁴⁹⁾,26⁽⁵⁰⁾-tetraene-51,52,53,54,55-pentone [ No CAS ]
(21E,23E,25E,26E,36R,37S,38R,39R,41S,43S,45R,46S,47R,48R,57R)-47,57-dihydroxy-45-[2-(2-hydroxyethoxy)ethoxy]-46-[(1R)-2-[(1S,3R,4R)-4-[2-(2-hydroxyethoxy)ethoxy]-3-methoxy-cyclohexyl]-1-methyl-ethyl]-48-methoxy-36,37,38,39,49,50-hexamethyl-68,69-dioxa-58-azatricyclohexatriaconta-21,23,25⁽⁴⁹⁾,26⁽⁵⁰⁾-tetraene-51,52,53,54,55-pentone [ No CAS ]

Reference： [1]Patent: WO2019/241789,2019,A1

49
[ 159351-69-6 ]
(22E,24E,26E,27E,31R,32S,33R,34R,36S,38S,40S,41S,42S,43R,52R)-42,52-dihydroxy-41-[(1R)-2-[(1S,3R,4R)-4-(2-hydroxyethoxy)-3-methoxy-cyclohexyl]-1-methyl-ethyl]-40,43-dimethoxy-31,32,33,34,44,45-hexamethyl-62,63-dioxa-53-azatricyclohexatriaconta-22,24,26⁽⁴⁴⁾,27⁽⁴⁵⁾-tetraene-46,47,48,49,50-pentone [ No CAS ]

Yield	Reaction Conditions	Operation in experiment
38%	With titanium(IV) isopropylate; In dichloromethane; at 20℃;	To a solution of <strong>[159351-69-6]everolimus</strong> (1 g, 1.04 mmol) in DCM (50 mL) was added Ti(OiPr)4 (0.89 g, 3.13 mmol) dropwise at rt. The reaction mixture turned pale yellow. After 30 min, the solution was poured into a separatory funnel containing a heterogeneous mixture of 1N HCl (50 mL) and EtOAc(50 mL). The organic layer was sequentially washed with saturated aqueous NaHCO3 (30 mL), H2O (50 mL), brine (50 mL), dried over anhydrous Na2SO4, filtered and concentrated under vacuum. The residue was purified by silica gel chromatography (hexane: acetone= 2:1) to obtain (22E,24E,26E,27E,31R,32S,33R,34R,36S,38S,40S,41S,42S,43R,52R)-42,52-dihydroxy-41- [(1R)-2-[(1S,3R,4R)-4-(2-hydroxyethoxy)-3-methoxy-cyclohexyl]-1-methyl-ethyl]-40,43- dimethoxy-31,32,33,34,44,45-hexamethyl-62,63-dioxa-53-azatricyclohexatriaconta- 22,24,26(44),27(45)-tetraene-46,47,48,49,50-pentone (380 mg, 38% yield) as a light yellow solid. 1H NMR (400 MHz, DMSO-d6) d 6.44 (dt, J = 13.9, 10.3 Hz, 2H), 6.33- 6.06 (m, 3H), 5.54- 4.85 (m, 5H), 4.48 (t, J = 5.3 Hz, 1H), 4.10- 3.91 (m, 2H), 3.89- 3.79 (m, 1H), 3.62 (d, J = 11.5 Hz, 1H), 3.54- 3.41 (m, 5H), 3.38- 3.28 (m, 8H), 3.19 (dt, J = 11.9, 7.6 Hz, 4H), 3.10- 2.92 (m, 6H), 2.71 (t, J = 14.8 Hz, 1H), 2.43- 1.78 (m, 6H), 1.75- 1.44 (m, 10H), 1.38 - 0.90 (m, 14H), 0.89- 0.67 (m, 13H), 0.66- 0.56 (m, 1H).

Reference： [1]Patent: WO2019/241789,2019,A1 .Location in patent: Paragraph 00560-00561

50
[ 159351-69-6 ]
[ 111-46-6 ]
(21E,23E,25E,26E,34R,35S,36R,37R,39S,41S,44S,45R,46R,55R)-45,55-dihydroxy-43-[2-(2-hydroxyethoxy)ethoxy]-44-[(1R)-2-[(1S,3R,4R)-4-(2-hydroxyethoxy)-3-methoxy-cyclohexyl]-1-methyl-ethyl]-46-methoxy-34,35,36,37,47,48-hexamethyl-66,67-dioxa-56-azatricyclohexatriaconta-21,23,25⁽⁴⁷⁾,26⁽⁴⁸⁾-tetraene-49,50,51,52,53-pentone [ No CAS ]

Yield	Reaction Conditions	Operation in experiment
19%	With toluene-4-sulfonic acid; In tetrahydrofuran; water; at 20℃; for 2h;	A mixture of (22E,24E,26E,27E,31R,32S,33R,34R,35S,38S,39S,40S,42R,43R, 52R)-42,52-dihydroxy-40-[(1R)-2-[(1S,2R,3R)-3-(2-hydroxyethoxy)-2-methoxy-cyclohexyl] -1-methyl-ethyl]-39,43-dimethoxy-31,32,33,34,44,45-hexamethyl-62,63-dioxa-53- azatricyclohexatriaconta-22,24,26(44),27(45)-tetraene-46,47,48,49,50-pentone (0.5 g, 0.52 mmol), 2-(2-hydroxyethoxy)ethanol (2.77 g, 26.09 mmol) and p-toluenesulfonic acid hydrate (0.54 g, 3.13 mmol) in THF (6 mL) was stirred at 20 C for 2 hours. The mixture was poured into ice cold saturated NaHCO3 (30 mL) and extracted with EtOAc (50 mL × 3). The organic layers were combined and then washed with water and brine, then concentrated in vacuo. The residue was purified by reverse phase chromatography (CH3CN/pure water = 1:1) to afford (21E,23E,25E,26E,34R,35S,36R,37R,38S,41S,43S,45R,46R,55R)-45,55-dihydroxy-42-[2-(2- hydroxyethoxy)ethoxy]-43-[(1R)-2-[(1S,2R,3R)-3-(2-hydroxyethoxy)-2-methoxy- cyclohexyl]-1-methyl-ethyl]-46-methoxy-34,35,36,37,47,48-hexamethyl-66,67-dioxa-56- azatricyclohexatriaconta-21,23,25(47),26(48)-tetraene-49,50,51,52,53-pentone (I-23: 0.1 g, 19% yield) as a white solid. ESI-MS (EI+, m/z): 1054.4 [M+Na]+.1H NMR (500 MHz, CDCl3) d 6.63- 5.86 (m, 4H), 5.70- 5.00 (m, 4H), 4.87- 4.15 (m, 2H), 4.02- 3.54 (m, 14H), 3.50- 3.26 (m, 11H), 3.24- 3.02 (m, 3H), 2.76- 2.46 (m, 3H), 2.38- 1.86 (m, 8H), 1.84- 1.54 (m, 14H), 1.47 (m, 3H), 1.25 (m, 6H), 1.00 (m, 16H), 0.69 (dt, J = 34.1, 12.1 Hz, 1H).

Reference： [1]Patent: WO2019/241789,2019,A1 .Location in patent: Paragraph 00349-00353

51
[ 159351-69-6 ]
[ 111-46-6 ]
(21E,23E,25E,26E,34R,35S,36R,37R,39S,41S,43R,44S,45R,46R,55R)-45,55-dihydroxy-43-[2-(2-hydroxyethoxy)ethoxy]-44-[(1R)-2-[(1S,3R,4R)-4-(2-hydroxyethoxy)-3-methoxy-cyclohexyl]-1-methyl-ethyl]-46-methoxy-34,35,36,37,47,48-hexamethyl-66,67-dioxa-56-azatricyclohexatriaconta-21,23,25⁽⁴⁷⁾,26⁽⁴⁸⁾-tetraene-49,50,51,52,53-pentone [ No CAS ]
(21E,23E,25E,26E,34R,35S,36R,37R,39S,41S,43S,44S,45R,46R,55R)-45,55-dihydroxy-43-[2-(2-hydroxyethoxy)ethoxy]-44-[(1R)-2-[(1S,3R,4R)-4-(2-hydroxyethoxy)-3-methoxy-cyclohexyl]-1-methyl-ethyl]-46-methoxy-34,35,36,37,47,48-hexamethyl-66,67-dioxa-56-azatricyclohexatriaconta-21,23,25⁽⁴⁷⁾,26⁽⁴⁸⁾-tetraene-49,50,51,52,53-pentone [ No CAS ]

Yield	Reaction Conditions	Operation in experiment
14 mg; 15 mg	With toluene-4-sulfonic acid; In tetrahydrofuran; at 0 - 25℃; for 3.5h;Inert atmosphere;	A solution of <strong>[159351-69-6]everolimus</strong> (1 g, 1.04 mmol) in THF (15 mL) was degassed with N2. p-toluenesulfonic acid (0.895 g, 5.20 mmol) was added at 0 C followed by 2-(2- hydroxyethoxy)ethanol (2.8 mL). The resulting mixture was stirred at 0C for 0.5 h under N2, then at 25C for 3 h. The reaction was poured into sat.NaHCO3 (40 mL), extracted with EtOAc (30 mL), washed with water (30 mL × 2) and brine (40 mL) then dried over anhydrous Na2SO4, filtered and concentrated. The resulting residue was purified by normal phase silica gel chromatography (MeOH: DCM = 1:15) then reverse-phase chromatography (C18, CH3CN: H2O= 7: 3) to obtain (21E,23E,25E,26E,34R,35S,36R,37R,39S,41S,44S,45R,46R,55R)- 45,55-dihydroxy-43-[2-(2-hydroxyethoxy)ethoxy]-44-[(1R)-2-[(1S,3R,4R)-4-(2- hydroxyethoxy)-3-methoxy-cyclohexyl]-1-methyl-ethyl]-46-methoxy-34,35,36,37,47,48- hexamethyl-66,67-dioxa-56-azatricyclohexatriaconta-21,23,25(47),26(48)-tetraene- 49,50,51,52,53-pentone (0.43 g, 40% yield) as a light yellow solid. ESI-MS (EI+, m/z):1053.9 [M+Na]+. 1H NMR (400 MHz, CDCl3) d6.47- 5.85 (m, 4H), 5.40 (ddd, J = 99.7, 51.9, 29.1 Hz, 4H), 4.80 (d, J = 22.0 Hz, 1H), 4.23 (d, J = 42.5 Hz, 1H), 4.05- 3.54 (m, 13H), 3.52- 3.01 (m, 14H), 2.67 (ddd, J = 46.8, 27.3, 6.8 Hz, 4H), 2.17 (dd, J = 82.0, 45.9 Hz, 6H), 1.70 (dt, J = 21.0, 15.8 Hz, 12H), 1.34 (dd, J = 105.2, 26.3 Hz, 11H), 1.15- 0.79 (m, 18H), 0.76- 0.64 (m, 1H). 90 mg of (21E,23E,25E,26E,34R,35S,36R,37R,39S,41S,44S,45R,46R,55R)- 45,55-dihydroxy-43-[2-(2-hydroxyethoxy)ethoxy]-44-[(1R)-2-[(1S,3R,4R)-4-(2- hydroxyethoxy)-3-methoxy-cyclohexyl]-1-methyl-ethyl]-46-methoxy-34,35,36,37,47,48- hexamethyl-66,67-dioxa-56-azatricyclohexatriaconta-21,23,25(47),26(48)-tetraene- 49,50,51,52,53-pentone was purified via prep chiral HPLC and the resulting epimers were obtained: (21E,23E,25E,26E,34R,35S,36R,37R,39S,41S,43R,44S,45R,46R,55R)-45,55- dihydroxy-43-[2-(2-hydroxyethoxy)ethoxy]-44-[(1R)-2-[(1S,3R,4R)-4-(2-hydroxyethoxy)-3- methoxy-cyclohexyl]-1-methyl-ethyl]-46-methoxy-34,35,36,37,47,48-hexamethyl-66,67- dioxa-56-azatricyclohexatriaconta-21,23,25(47),26(48)-tetraene-49,50,51,52,53-pentone (I- 116: 14 mg, 16% yield) and (21E,23E,25E,26E,34R,35S,36R,37R,39S,41S,43S,44S,45R,46R,55R)-45,55-dihydroxy-43- [2-(2-hydroxyethoxy)ethoxy]-44-[(1R)-2-[(1S,3R,4R)-4-(2-hydroxyethoxy)-3-methoxy- cyclohexyl]-1-methyl-ethyl]-46-methoxy-34,35,36,37,47,48-hexamethyl-66,67-dioxa-56- azatricyclohexatriaconta-21,23,25(47),26(48)-tetraene-49,50,51,52,53-pentone (I-117: 15 mg, 17% yield), both as white solids. The chiral separation method is listed below: I-116: ESI-MS (EI+, m/z):1054.0 [M+Na]+. 1H NMR (500 MHz, , CDCl3) d 6.42 - 5.90 (m, 4H), 5.79 (ddd, J = 51.6, 30.9, 16.6 Hz, 1H), 5.54- 5.08 (m, 4H), 5.03- 4.88 (m, 1H), 4.74 (d, J = 61.6 Hz, 1H), 4.28 (dd, J = 57.6, 29.0 Hz, 2H), 3.99 (dd, J = 26.5, 6.0 Hz, 1H), 3.89- 3.55 (m, 12H), 3.54- 2.96 (m, 15H), 2.87- 2.47 (m, 4H), 2.38- 1.92 (m, 8H), 1.86- 1.67 (m, 11H), 1.51- 1.30 (m, 6 H), 1.14- 0.80 (m, 18H), 0.76- 0.64 (m, 1H). I-117: ESI-MS (EI+, m/z):1053.9 [M+Na]+. 1H NMR (500 MHz, , CDCl3) d 6.41 - 6.22 (m, 2H), 6.13 (dd, J = 15.1, 10.1 Hz, 1H), 5.94 (dd, J = 22.3, 10.8 Hz, 1H), 5.52 (dt, J = 18.4, 9.2 Hz, 1H), 5.41 (d, J = 9.9 Hz, 1H), 5.27 (d, J = 5.3 Hz, 1H), 5.12 (dt, J = 46.3, 5.6 Hz, 1H), 4.83 (s, 1H), 4.23- 4.14 (m, 1H), 3.91- 3.52 (m, 15H), 3.49- 3.25 (m, 12H), 3.23 - 3.03 (m, 3H), 2.94- 2.80 (m, 1H), 2.65 (ddd, J = 23.4, 16.9, 6.0 Hz, 3H), 2.39- 2.15 (m, 2H), 2.16- 1.85 (m, 5H), 1.82- 1.64 (m, 10H), 1.47 (dd, J = 26.8, 15.9 Hz, 5H), 1.38- 1.16 (m, 6H), 1.10 (d, J = 6.8 Hz, 3H), 1.07- 1.03 (m, 3H), 1.00 (t, J = 6.8 Hz, 3H), 0.94 (d, J = 6.6 Hz, 3H), 0.88 (ddd, J = 34.0, 16.9, 5.0 Hz, 6H), 0.71 (dd, J = 23.9, 11.8 Hz, 1H).

Reference： [1]Patent: WO2019/241789,2019,A1 .Location in patent: Paragraph 00385-00390

52
[ 159351-69-6 ]
[ 112-27-6 ]
(21E,23E,25E,26E,36R,37S,38R,39R,41S,43S,46S,47R,48R,57R)-47,57-dihydroxy-45-[2-[2-(2-hydroxyethoxy)ethoxy]ethoxy]-46-[(1R)-2-[(1S,3R,4R)-4-(2-hydroxyethoxy)-3-methoxy-cyclohexyl]-1-methyl-ethyl]-48-methoxy-36,37,38,39,49,50-hexamethyl-68,69-dioxa-58-azatricyclohexatriaconta-21,23,25⁽⁴⁹⁾,26⁽⁵⁰⁾-tetraene-51,52,53,54,55-pentone [ No CAS ]

Yield	Reaction Conditions	Operation in experiment
28%	With toluene-4-sulfonic acid; In tetrahydrofuran; water; at 20℃; for 2h;	A mixture of <strong>[159351-69-6]everolimus</strong> (3.92 g, 26.09 mmol) and p-toluenesulfonic acid hydrate (0.45 g, 2.61 mmol) in THF (10 mL) was stirred at 20 C for 2 hours. The mixture was poured into ice cold saturated NaHCO3 (30 mL) and extracted with EtOAc (50 mL × 3). The organic layers were combined and then washed with water and brine, then concentrated in vacuo. The residue was purified by reversed phase chromatography (CH3CN/pure water = 1:1) to afford (21E,23E,25E,26E,36R,37S,38R,39R,40S,43S,45S,47R,48R,57R)-47,57-dihydroxy-44-[2-[2- (2-hydroxyethoxy)ethoxy]ethoxy]-45-[(1R)-2-[(1S,2R,3R)-3-(2-hydroxyethoxy)-2-methoxy- cyclohexyl]-1-methyl-ethyl]-48-methoxy-36,37,38,39,49,50-hexamethyl-68,69-dioxa-58- azatricyclohexatriaconta-21,23,25(49),26(50)-tetraene-51,52,53,54,55-pentone (I-32: 0.155 g, 28% yield) as a white solid. ESI-MS (EI+, m/z): 1098.4 [M+Na]+. 1H NMR (500 MHz, CDCl3) d 6.49- 5.83 (m, 4H), 5.67- 5.35 (m, 2H), 5.33- 5.01 (m, 2H), 4.92- 4.08 (m, 2H), 4.05- 3.51 (m, 17H), 3.52- 3.23 (m, 11H), 3.23- 3.01 (m, 3H), 2.66 (m, 4H), 2.40- 1.95 (m, 5H), 1.95- 1.55 (m, 17H), 1.52- 1.13 (m, 9H), 1.13- 0.79 (m, 16H), 0.71 (dd, J = 23.8, 11.9 Hz, 1H).

Reference： [1]Patent: WO2019/241789,2019,A1 .Location in patent: Paragraph 00354-00355

53
[ 159351-69-6 ]
[ 112-27-6 ]
(21E,23E,25E,26E,36R,37S,38R,39R,41S,43S,45R,46S,47R,48R,57R)-47,57-dihydroxy-45-[2-[2-(2-hydroxyethoxy)ethoxy]ethoxy]-46-[(1R)-2-[(1S,3R,4R)-4-(2-hydroxyethoxy)-3-methoxy-cyclohexyl]-1-methyl-ethyl]-48-methoxy-36,37,38,39,49,50-hexamethyl-68,69-dioxa-58-azatricyclohexatriaconta-21,23,25⁽⁴⁹⁾,26⁽⁵⁰⁾-tetraene-51,52,53,54,55-pentone [ No CAS ]
(21E,23E,25E,26E,36R,37S,38R,39R,41S,43S,45S,46S,47R,48R,57R)-47,57-dihydroxy-45-[2-[2-(2-hydroxyethoxy)ethoxy]ethoxy]-46-[(1R)-2-[(1S,3R,4R)-4-(2-hydroxyethoxy)-3-methoxy-cyclohexyl]-1-methyl-ethyl]-48-methoxy-36,37,38,39,49,50-hexamethyl-68,69-dioxa-58-azatricyclohexatriaconta-21,23,25⁽⁴⁹⁾,26⁽⁵⁰⁾-tetraene-51,52,53,54,55-pentone [ No CAS ]

Yield	Reaction Conditions	Operation in experiment
33 mg; 37 mg	With toluene-4-sulfonic acid; In tetrahydrofuran; water; for 2h;	A mixture of <strong>[159351-69-6]everolimus</strong> (0.5 g, 0.522 mmol), 2-[2-(2- hydroxyethoxy)ethoxy]ethanol (3.92 g, 26.09 mmol) and p-toluenesulfonic acid monohydrate (0.45 g, 2.61 mmol) in THF (10 mL) was stirred at 20 C for 2 h. The mixture was then poured into ice cold sat. NaHCO3 (30 mL) and extracted with EtOAc (50 mL × 3). The combined organic layers were washed with water and brine then dried over anhydrous Na2SO4, filtered and concentrated. The reslting residue was purified via reverse phase chromatography (C18, CH3CN:H2O= 6.5: 3.5) to afford (21E,23E,25E,26E,36R,37S,38R,39R,41S,43S,46S,47R,48R,57R)-47,57-dihydroxy-45-[2-[2- (2-hydroxyethoxy)ethoxy]ethoxy]-46-[(1R)-2-[(1S,3R,4R)-4-(2-hydroxyethoxy)-3-methoxy- cyclohexyl]-1-methyl-ethyl]-48-methoxy-36,37,38,39,49,50-hexamethyl-68,69-dioxa-58- azatricyclohexatriaconta-21,23,25(49),26(50)-tetraene-51,52,53,54,55-pentone (0.155 g, 28%) as white solid. ESI-MS (EI+, m/z):1098.4 [M+Na]+. 1H NMR (500 MHz, CDCl3) d 6.42- 5.87 (m, 4H), 5.63- 5.34 (m, 2H), 5.32- 5.00 (m, 2H), 4.85 (s, 1H), 4.35- 4.09 (m, 1H), 4.05- 3.49 (m, 18H), 3.49- 3.01 (m, 14H), 2.66 (dddd, J = 31.3, 24.8, 21.2, 13.0 Hz, 4H), 2.33 (d, J = 12.0 Hz, 2H), 2.06 (dd, J = 39.9, 10.6 Hz, 3H), 1.77- 1.53 (m, 13H), 1.51- 1.14 (m, 10H), 1.14- 0.81 (m, 18H), 0.71 (dd, J = 23.8, 11.9 Hz, 1H). 170 mg of (21E,23E,25E,26E,36R,37S,38R,39R,41S,43S,46S,47R,48R,57R)- 47,57-dihydroxy-45-[2-[2-(2-hydroxyethoxy)ethoxy]ethoxy]-46-[(1R)-2-[(1S,3R,4R)-4-(2- hydroxyethoxy)-3-methoxy-cyclohexyl]-1-methyl-ethyl]-48-methoxy-36,37,38,39,49,50- hexamethyl-68,69-dioxa-58-azatricyclohexatriaconta-21,23,25(49),26(50)-tetraene- 51,52,53,54,55-pentone purified via prep chiral HPLC and the resulting epimers purified via silica gel chromatography (hexane:DCM:EtOAc:MeOH = 3:3:1:0.8) to obtain (21E,23E,25E,26E,36R,37S,38R,39R,41S,43S,45S,46S,47R,48R,57R)-47,57-dihydroxy-45- [2-[2-(2-hydroxyethoxy)ethoxy]ethoxy]-46-[(1R)-2-[(1S,3R,4R)-4-(2-hydroxyethoxy)-3- methoxy-cyclohexyl]-1-methyl-ethyl]-48-methoxy-36,37,38,39,49,50-hexamethyl-68,69- dioxa-58-azatricyclohexatriaconta-21,23,25(49),26(50)-tetraene-51,52,53,54,55-pentone (I- 110: 37 mg, 21% yield) and (21E,23E,25E,26E,36R,37S,38R,39R,41S,43S,45R,46S,47R,48R,57R)-47,57-dihydroxy-45- [2-[2-(2-hydroxyethoxy)ethoxy]ethoxy]-46-[(1R)-2-[(1S,3R,4R)-4-(2-hydroxyethoxy)-3- methoxy-cyclohexyl]-1-methyl-ethyl]-48-methoxy-36,37,38,39,49,50-hexamethyl-68,69- dioxa-58-azatricyclohexatriaconta-21,23,25(49),26(50)-tetraene-51,52,53,54,55-pentone (I- 109: 33 mg, 19% yield), both as white solids. Chiral analysis method: C I-110: ESI-MS (EI+, m/z):1098.0 [M+Na]+. 1H NMR (500 MHz, CDCl3) d 6.41- 6.18 (m, 2H), 6.12 (dd, J = 15.0, 10.3 Hz, 1H), 5.93 (dd, J = 33.7, 10.6 Hz, 1H), 5.47 (ddd, J = 33.0, 20.8, 9.5 Hz, 2H), 5.26 (d, J = 5.6 Hz, 1H), 5.13 (dt, J = 21.8, 10.8 Hz, 1H), 4.88 (s, 1H), 4.19 (t, J = 9.3 Hz, 1H), 3.94- 3.52 (m, 19H), 3.49- 3.25 (m, 12H), 3.24- 3.02 (m, 3H), 2.76 (ddd, J = 26.2, 16.6, 10.3 Hz, 3H), 2.57 (dd, J = 17.0, 6.3 Hz, 1H), 2.29 (t, J = 26.2 Hz, 2H), 2.16- 1.85 (m, 6H), 1.74- 1.53 (m, 10H), 1.53- 1.16 (m, 9H), 1.15- 1.01 (m, 8H), 1.01- 0.82 (m, 10H), 0.71 (dd, J = 23.9, 12.0 Hz, 1H). I-109: ESI-MS (EI+, m/z):1098.0 [M+Na]+. 1H NMR (500 MHz, CDCl3) d 6.43- 5.90 (m, 4H), 5.62- 5.02 (m, 5H), 4.24 (d, J = 63.6 Hz, 1H), 3.97 (dd, J = 21.5, 6.8 Hz, 1H), 3.86- 3.50 (m, 18H), 3.45- 3.01 (m, 14H), 2.73- 2.46 (m, 3H), 2.39- 1.94 (m, 6H), 1.91- 1.69 (m, 10H), 1.50- 1.31 (m, 12H), 1.16- 0.85 (m, 18H), 0.69 (d, J = 11.7 Hz, 1H).

Reference： [1]Patent: WO2019/241789,2019,A1 .Location in patent: Paragraph 00405-00410

54
[ 5617-32-3 ]
[ 159351-69-6 ]
(21E,23E,25E,26E,44R,45S,46R,47R,49S,51S,54S,55R,56R,65R)-55,65-dihydroxy-53-[2-[2-[2-[2-[2-[2-(2-hydroxyethoxy)ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]-54-[(1R)-2-[(1S,3R,4R)-4-(2-hydroxyethoxy)-3-methoxy-cyclohexyl]-1-methyl-ethyl]-56-methoxy-44,45,46,47,57,58-hexamethyl-76,77-dioxa-66-azatricyclohexatriaconta-21,23,25⁽⁵⁷⁾,26⁽⁵⁸⁾-tetraene-59,60,61,62,63-pentone [ No CAS ]

Yield	Reaction Conditions	Operation in experiment
14%	With toluene-4-sulfonic acid; In tetrahydrofuran; water; at 22℃; for 18h;Inert atmosphere;	To a solution of <strong>[159351-69-6]everolimus</strong> (0.5 g, 0.52 mmol) and 2-[2-[2-[2-[2-[2-(2- hydroxyethoxy)ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethanol (3.41 g, 10.44 mmol) in THF (15 mL) was added 4-methylbenzenesulfonic acid hydrate (0.52 g, 2.73 mmol) slowly. The resulting mixture was stirred at 22 C for 18 h under N2 then quenched with aq. NaHCO3 (30 mL) and extracted with EtOAc (60 mL× 3). The combined organic layers were washed with water (50 mL), brine (50 mL), dried over anhydrous Na2SO4, filtered, and concentrated under vacuum. The residue was purified by reverse phase chromatography (C18, CH3CN:H2O = 70:30) to afford (21E,23E,25E,26E,44R,45S,46R,47R,49S,51S,54S,55R,56R,65R)-55,65- dihydroxy-53-[2-[2-[2-[2-[2-[2-(2-hydroxyethoxy)ethoxy]ethoxy]ethoxy]ethoxy]ethoxy] ethoxy]-54-[(1R)-2-[(1S,3R,4R)-4-(2-hydroxyethoxy)-3-methoxy-cyclohexyl]-1-methyl- ethyl]-56-methoxy-44,45,46,47,57,58-hexamethyl-76,77-dioxa-66-azatricyclohexatriaconta- 21,23,25(57),26(58)-tetraene-59,60,61,62,63-pentone (I-99: 93 mg, 14% yield) as a white solid. ESI-MS (EI+, m/z): 1274.9 [M+Na]+. 1H NMR (500 MHz, CDCl3) d 6.42- 5.82 (m, 4H), 5.64- 5.06 (m, 4H), 4.81 (s, 1H), 4.30- 4.08 (m, 1H), 3.81- 3.53 (m, 35H), 3.46- 3.26 (m, 12H), 3.22- 3.05 (m, 4H), 2.76- 2.65 (m, 2H), 2.39- 2.22 (m, 2H), 2.14- 1.97 (m, 3H), 1.75- 1.55 (m, 13H), 1.52- 1.39 (m, 4H), 1.30- 1.13 (m, 6H), 1.08- 0.82 (m, 17H), 0.76- 0.65 (m, 1H).

Reference： [1]Patent: WO2019/241789,2019,A1 .Location in patent: Paragraph 00530-00531

55
[ 111-77-3 ]
[ 159351-69-6 ]
(22E,24E,26E,27E,35R,36S,37R,38R,40S,42S,44R,45S,46R,47R,56R)-46,56-dihydroxy-45-[(1R)-2-[(1S,3R,4R)-4-(2-hydroxyethoxy)-3-methoxy-cyclohexyl]-1-methyl-ethyl]-47-methoxy-44-[2-(2-methoxyethoxy)ethoxy]-35,36,37,38,48,49-hexamethyl-66,67-dioxa-57-azatricyclohexatriaconta-22,24,26⁽⁴⁸⁾,27⁽⁴⁹⁾-tetraene-50,51,52,53,54-pentone [ No CAS ]
(22E,24E,26E,27E,35R,36S,37R,38R,40S,42S,44S,45S,46R,47R,56R)-46,56-dihydroxy-45-[(1R)-2-[(1S,3R,4R)-4-(2-hydroxyethoxy)-3-methoxy-cyclohexyl]-1-methyl-ethyl]-47-methoxy-44-[2-(2-methoxyethoxy)ethoxy]-35,36,37,38,48,49-hexamethyl-66,67-dioxa-57-azatricyclohexatriaconta-22,24,26⁽⁴⁸⁾,27⁽⁴⁹⁾-tetraene-50,51,52,53,54-pentone [ No CAS ]

Yield	Reaction Conditions	Operation in experiment
15 mg; 17 mg	With HND-8; In tetrahydrofuran; at 50℃; for 4h;Inert atmosphere;	A mixture of <strong>[159351-69-6]everolimus</strong> (5 g, 5.22 mmol) and 2-(2-methoxyethoxy)ethanol (15 mL) in THF (80 mL) was degassed with N2 then heated at 50 C. HND-8 (600 mg) was added and the resulting mixture stirred at 50C for 4 h under N2 then filtered and diluted with EtOAc. After concentration, the residue was purified by reverse phase chromatography (C18, CH3CN: H2O from 0%-100% yield) to afford (22E,24E,26E,27E,35R,36S,37R,38R,40S,42S,45S,46R,47R,56R)-46,56-dihydroxy-45- [(1R)-2-[(1S,3R,4R)-4-(2-hydroxyethoxy)-3-methoxy-cyclohexyl]-1-methyl-ethyl]-47- methoxy-44-[2-(2-methoxyethoxy)ethoxy]-35,36,37,38,48,49-hexamethyl-66,67-dioxa-57- azatricyclohexatriaconta-22,24,26(48),27(49)-tetraene-50,51,52,53,54-pentone (1.5 g, 27% yield) as a white solid. ESI-MS (EI+, m/z):1068.4 [M+Na]+. 1H NMR (400 MHz, CDCl3) d 6.40- 5.85 (m, 4H), 5.56- 5.36 (m, 2H), 5.21 (ddd, J = 16.0, 11.7, 5.6 Hz, 2H), 4.21 (dd, J = 25.8, 15.5 Hz, 2H), 3.94- 3.26 (m, 28H), 3.25- 3.02 (m, 4H), 2.76- 2.40 (m, 3H), 2.34 (d, J = 13.3 Hz, 2H), 2.19- 2.06 (m, 2H), 2.01- 1.67 (m, 13H), 1.54- 1.30 (m, 7H), 1.15- 0.81 (m, 18H), 0.72 (dd, J = 23.1, 11.7 Hz, 1H). 120 mg of (22E,24E,26E,27E,35R,36S,37R,38R,40S,42S,45S,46R,47R,56R)- 46,56-dihydroxy-45-[(1R)-2-[(1S,3R,4R)-4-(2-hydroxyethoxy)-3-methoxy-cyclohexyl]-1- methyl-ethyl]-47-methoxy-44-[2-(2-methoxyethoxy)ethoxy]-35,36,37,38,48,49-hexamethyl- 66,67-dioxa-57-azatricyclohexatriaconta-22,24,26(48),27(49)-tetraene-50,51,52,53,54- pentone was purified via prep chiral HPLC to obtain (22E,24E,26E,27E,35R,36S,37R,38R,40S,42S,44S,45S,46R,47R,56R)-46,56-dihydroxy-45- [(1R)-2-[(1S,3R,4R)-4-(2-hydroxyethoxy)-3-methoxy-cyclohexyl]-1-methyl-ethyl]-47- methoxy-44-[2-(2-methoxyethoxy)ethoxy]-35,36,37,38,48,49-hexamethyl-66,67-dioxa-57- azatricyclohexatriaconta-22,24,26(48),27(49)-tetraene-50,51,52,53,54-pentone (I-105: 17 mg, 20% yield) and (22E,24E,26E,27E,35R,36S,37R,38R,40S,42S,44R,45S,46R,47R,56R)-46,56- dihydroxy-45-[(1R)-2-[(1S,3R,4R)-4-(2-hydroxyethoxy)-3-methoxy-cyclohexyl]-1-methyl- ethyl]-47-methoxy-44-[2-(2-methoxyethoxy)ethoxy]-35,36,37,38,48,49-hexamethyl-66,67- dioxa-57-azatricyclohexatriaconta-22,24,26(48),27(49)-tetraene-50,51,52,53,54-pentone (I- 104: 15 mg, 16% yield), both as white solids. Chiral separation method: Column : CHIRALPAK IC Column size : 2.5 cm I.D. × 25 cm L, 10mm Sample solution: 14 mg/ml in Mobile phase Injection : 15 ml Mobile phase : Hexane/EtOH=50/50(V/V) Flow rate : 60 ml/min Wave length : UV 254 nm Temperature : 35 C I-105: ESI-MS (EI+, m/z):1068.4 [M+Na]+. 1H NMR (400 MHz, CDCl3) d 6.42- 6.19 (m, 2H), 6.13 (dd, J = 15.1, 10.0 Hz, 1H), 5.91 (dd, J = 33.0, 10.5 Hz, 1H), 5.56- 5.38 (m, 2H), 5.27 (d, J = 5.0 Hz, 1H), 5.15 (dt, J = 15.2, 7.6 Hz, 1H), 4.76 (s, 1H), 4.18 (d, J = 5.6 Hz, 1H), 3.93- 3.25 (m, 30H), 3.24- 3.03 (m, 3H), 2.72 (dd, J = 16.7, 5.6 Hz, 2H), 2.57 (dd, J = 16.8, 6.5 Hz, 1H), 2.34 (d, J = 13.9 Hz, 2H), 2.13- 1.84 (m, 6H), 1.82- 1.67 (m, 7H), 1.47 (dd, J = 24.1, 16.7 Hz, 4H), 1.25 (ddd, J = 24.1, 20.2, 10.0 Hz, 7H), 1.14- 0.81 (m, 18H), 0.72 (dd, J = 23.9, 12.1 Hz, 1H). I-104: ESI-MS (EI+, m/z):1068.4 [M+Na]+. 1H NMR (400 MHz, CDCl3) d 6.19 (m, 4H), 5.56- 5.36 (m, 2H), 5.28- 5.07 (m, 2H), 4.83 (d, J = 4.9 Hz, 4H), 4.28 (s, 1H), 4.21 - 4.09 (m, 1H), 4.04- 3.51 (m, 15H), 3.46- 3.29 (m, 11H), 3.27- 2.91 (m, 5H), 2.76- 2.42 (m, 3H), 2.31 (d, J = 11.3 Hz, 2H), 2.18- 1.70 (m, 13H), 1.53- 1.19 (m, 8H), 1.16- 0.84 (m, 18H), 0.76- 0.60 (m, 1H).

Reference： [1]Patent: WO2019/241789,2019,A1 .Location in patent: Paragraph 00420-00425

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Code	Phrase
H400	Very toxic to aquatic life
H401	Toxic to aquatic life
H402	Harmful to aquatic life
H410	Very toxic to aquatic life with long-lasting effects
H411	Toxic to aquatic life with long-lasting effects
H412	Harmful to aquatic life with long-lasting effects
H413	May cause long-lasting harmful effects to aquatic life
H420	Harms public health and the environment by destroying ozone in the upper atmosphere

[ CAS No. 159351-69-6 ] {[proInfo.proName]}

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[ 159351-69-6 ] Synthesis Path-Upstream 1~7

[ 159351-69-6 ] Synthesis Path-Downstream 1~55

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