* 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.
With dicyclohexyl-carbodiimide In dichloromethane at 15 - 20℃;
Step2: 2,5-Dioxopyrrolidine-1-yl 2-(fert-butoxycarbonylamino)acetate. /V-Boc-glycine (7.31 g, 41 .7 mmol) was dissolved in 100 mL of DCM and to the cooled (15°C) solution /V-hydroxysuccinimide (5.28 g, 45.9 mmol) was added. Λ/,Λ/'-dicyclohexylcarbodiimide (9.47 g, 45.9 mmol) was added to the formed suspension under vigorous stirring. After a few seconds, a cloudy white suspension formed, the mixture was allowed to reach room temperature and stirred for 1 h. It was subsequently filtrated over celite, washed with 50m L saturated sodium - - bicarbonate, dried over sodium sulfate and concentrated in vacuo to yield a crystalline powder. Yield: 7.02 g, 61 .8percent 1 H NMR (400 MHz, DMSO -cfe) : δ 1 .38 (s, 9H), 2.80 (s, 4H), 4.07 (d, J = 6 Hz, 2H), 7.43 (br s, 1 H). MS (ESI) m/z 567.2 [2M + Na]+.
61.8%
With dicyclohexyl-carbodiimide In dichloromethane at 15℃;
N-Boc-glycine (7.31 g, 41.7 mmol) was dissolved in 100 mL of DCM and to the cooled (15° C.) solution N-hydroxysuccinimide (5.28 g, 45.9 mmol) was added. N,N'-dicyclohexylcarbodiimide (9.47 g, 45.9 mmol) was added to the formed suspension under vigorous stirring. After a few seconds, a cloudy white suspension formed, the mixture was allowed to reach room temperature and stirred for 1 h. It was subsequently filtrated over celite, washed with 50 mL saturated sodium bicarbonate, dried over sodium sulfate and concentrated in vacuo to yield a crystalline powder. Yield: 7.02 g, 61.8percent 1H NMR (400 MHz, DMSO-d6): δ 1.38 (s, 9H), 2.80 (s, 4H), 4.07 (d, J=6 Hz, 2H), 7.43 (br s, 1H). MS (ESI) m/z 567.2 [2M+Na]+.
Reference：
[1] Chemistry - A European Journal, 2016, vol. 22, # 52, p. 18865 - 18872
[2] Journal of the Chemical Society - Perkin Transactions 1, 1998, # 15, p. 2443 - 2449
[3] Chemical communications (Cambridge, England), 2003, # 23, p. 2870 - 2871
[4] ACS Medicinal Chemistry Letters, 2013, vol. 4, # 4, p. 402 - 407
[5] Helvetica Chimica Acta, 2004, vol. 87, # 5, p. 1077 - 1089
[6] Bioconjugate Chemistry, 2010, vol. 21, # 9, p. 1642 - 1655
[7] Patent: WO2013/107820, 2013, A1, . Location in patent: Page/Page column 42; 43
[8] Patent: US2014/357650, 2014, A1, . Location in patent: Paragraph 0265; 0268; 0269
[9] Journal of the Chemical Society, Dalton Transactions: Inorganic Chemistry (1972-1999), 1984, p. 2305 - 2308
[10] Journal of the American Chemical Society, 1993, vol. 115, # 10, p. 4228 - 4245
[11] Pharmaceutical Chemistry Journal, 1992, vol. 26, # 9-10, p. 753 - 756[12] Khimiko-Farmatsevticheskii Zhurnal, 1992, vol. 26, # 9-10, p. 72 - 74
[13] Journal of the American Chemical Society, 1996, vol. 118, # 29, p. 6975 - 6985
[14] Nucleosides and Nucleotides, 1998, vol. 17, # 9-11, p. 2135 - 2141
[15] Chemistry - A European Journal, 2001, vol. 7, # 19, p. 4280 - 4295
[16] Bioorganic and Medicinal Chemistry Letters, 2009, vol. 19, # 12, p. 3220 - 3224
[17] Organic and Biomolecular Chemistry, 2015, vol. 13, # 13, p. 3950 - 3962
[18] Organic Process Research and Development, 2015, vol. 19, # 9, p. 1257 - 1262
[19] Journal of Molecular Biology, 2018, vol. 430, # 6, p. 842 - 852
2
[ 4530-20-5 ]
[ 3392-07-2 ]
Reference：
[1] Patent: WO2006/17352, 2006, A1, . Location in patent: Page/Page column 38
[2] Patent: US4782164, 1988, A,
Reference：
[1] J. Gen. Chem. USSR (Engl. Transl.), 1984, vol. 54, # 8, p. 1696 - 1698[2] Zhurnal Obshchei Khimii, 1984, vol. 54, # 8, p. 1904 - 1907
[3] Bulletin of the Academy of Sciences of the USSR, Division of Chemical Science (English Translation), 1984, vol. 33, p. 1067 - 1070[4] Izvestiya Akademii Nauk SSSR, Seriya Khimicheskaya, 1984, # 5, p. 1163 - 1165
6
[ 4530-20-5 ]
[ 107960-02-1 ]
[ 3392-07-2 ]
Reference：
[1] Journal of Organic Chemistry, 1987, vol. 52, # 12, p. 2364 - 2367
With dicyclohexyl-carbodiimide In tetrahydrofuran at 20℃; for 16h; Inert atmosphere;
86%
With dicyclohexyl-carbodiimide In dichloromethane for 2h; Cooling with ice;
85%
With dicyclohexyl-carbodiimide In 1,2-dimethoxyethane at 0℃;
83%
With dicyclohexyl-carbodiimide In tetrahydrofuran at 20℃; for 1h;
78%
With dicyclohexyl-carbodiimide at 0 - 20℃; for 8.5h;
4.2.1. General Synthetic Method for the Preparation of N-Hydroxysuccinimide Esters of Carboxylic and Amino Acids
General procedure: To a well-stirred solution of 17 mmol of acid in 50 mlTHF (or ethylacetate) was added 18 mmol HOSu, then thesolution was cooled to 0°C, and 18 mmol of DCCwas added, ensuring that the temperature did not exceed0 - + 5°C. Stirring continued for 30 min at + 5 °C and then 8h at room temperature. DCU was filtered off, the solutionwas evaporated. The resulting oil was triturated with hexaneto remove the excess of DCC, hexane was decanted. The oilwas dissolved in 30 ml of dichloromethane. The solution waskept at cool temperature (+5 - +8°C) for 24 hours, the reprecipitateof DCU was filtered off, the solution was washedwith 5% NaHCO3. Organic layer was dried using anhydrousMgSO4 or CaCl2, filtered, the solvent was evaporated. Productwas obtained as crystals or viscous yellow oil.
75%
With dmap; dicyclohexyl-carbodiimide In dichloromethane at 0 - 20℃; for 16.75h;
1.2
In the second step, 25 g of BOC-glycine prepared in the first step was dissolved in 300 mL of dichloromethane, and 23.92 g of N-hydroxysuccinimide and 1.75 g of 4-dimethylaminopyridine were added to lower the temperature of the reaction system. After 0 ° C, 200 mL of a dichloromethane solution containing 44.15 g of dicyclohexylcarbodiimide was added dropwise to the reaction system, and after the completion of the dropwise addition, the reaction was continued at 0 ° C for 45 minutes, and then the temperature of the system was gradually raised to room temperature to continue the reaction. After 16 hours; after completion of the reaction, the mixture was washed with 0.05 mol/L HCl solution (2×100 mL), and the organic phase was collected and dried over anhydrous sodium sulfate. The volume of ethyl acetate was crystallized at -20 ° C, and filtered to obtain a pale yellow solid product (BOC-glycine-OSu) 29.15 g, a yield of about 75%;
74%
With dmap; dicyclohexyl-carbodiimide In dichloromethane at 0 - 20℃;
68%
With dicyclohexyl-carbodiimide In tetrahydrofuran at 20℃;
68.8%
With dicyclohexyl-carbodiimide In tetrahydrofuran at 20℃;
61.8%
With dicyclohexyl-carbodiimide In dichloromethane at 15 - 20℃;
2 Step2: 2,5-Dioxopyrrolidine-1-yl 2-(fert-butoxycarbonylamino)acetate
Step2: 2,5-Dioxopyrrolidine-1-yl 2-(fert-butoxycarbonylamino)acetate. /V-Boc-glycine (7.31 g, 41 .7 mmol) was dissolved in 100 mL of DCM and to the cooled (15°C) solution /V-hydroxysuccinimide (5.28 g, 45.9 mmol) was added. Λ/,Λ/'-dicyclohexylcarbodiimide (9.47 g, 45.9 mmol) was added to the formed suspension under vigorous stirring. After a few seconds, a cloudy white suspension formed, the mixture was allowed to reach room temperature and stirred for 1 h. It was subsequently filtrated over celite, washed with 50m L saturated sodium - - bicarbonate, dried over sodium sulfate and concentrated in vacuo to yield a crystalline powder. Yield: 7.02 g, 61 .8% 1 H NMR (400 MHz, DMSO -cfe) : δ 1 .38 (s, 9H), 2.80 (s, 4H), 4.07 (d, J = 6 Hz, 2H), 7.43 (br s, 1 H). MS (ESI) m/z 567.2 [2M + Na]+.
61.8%
With dicyclohexyl-carbodiimide In dichloromethane at 15℃;
2 Step 2: 2,5-Dioxopyrrolidine-1-yl 2-(tert-butoxycarbonylamino)acetate
N-Boc-glycine (7.31 g, 41.7 mmol) was dissolved in 100 mL of DCM and to the cooled (15° C.) solution N-hydroxysuccinimide (5.28 g, 45.9 mmol) was added. N,N'-dicyclohexylcarbodiimide (9.47 g, 45.9 mmol) was added to the formed suspension under vigorous stirring. After a few seconds, a cloudy white suspension formed, the mixture was allowed to reach room temperature and stirred for 1 h. It was subsequently filtrated over celite, washed with 50 mL saturated sodium bicarbonate, dried over sodium sulfate and concentrated in vacuo to yield a crystalline powder. Yield: 7.02 g, 61.8% 1H NMR (400 MHz, DMSO-d6): δ 1.38 (s, 9H), 2.80 (s, 4H), 4.07 (d, J=6 Hz, 2H), 7.43 (br s, 1H). MS (ESI) m/z 567.2 [2M+Na]+.
With dicyclohexyl-carbodiimide
With dicyclohexyl-carbodiimide In tetrahydrofuran for 1h;
With dicyclohexyl-carbodiimide In N,N-dimethyl-formamide for 24h; Ambient temperature;
With dicyclohexyl-carbodiimide In dichloromethane for 1h; Ambient temperature;
With dicyclohexyl-carbodiimide
With N-(3-dimethylaminopropyl)-N-ethylcarbodiimide In 1,4-dioxane at 23℃; for 2h;
With dicyclohexyl-carbodiimide In 1,4-dioxane
With dicyclohexyl-carbodiimide In tetrahydrofuran at 0 - 20℃; for 2h;
With dicyclohexyl-carbodiimide In 1,4-dioxane at 20℃; Large scale;
With dicyclohexyl-carbodiimide at 0℃;
Chemical synthesis of AA-CAM derivatives
General procedure: The general scheme for the synthesis of AA-CAMderivatives is shown in Fig. S1 and the details ofchemical synthesis are provided in the SupplementaryMethods section. CAM [(1R,2R)-2-amino-1-(4--nitrophenyl)propane-1,3-diol)] was prepared asdescribed previously [23]. Amino acids with protectedα- and side-chain amino groups wereactivated by reaction with N-hydroxysuccinimide inthe presence of N,N′-dicyclohexylcarbodiimide at0 °C. The resulting succinimide-reactive esters wereused for the acylation of CAM in the presence ofdiisopropylethylamine as a base at room temperature.Subsequent deprotection was achieved bytreatment of the obtained amino-acid CAM derivativeswith trifluoroacetic acid and appropriate scavengers.Synthesized AA-CAM derivatives were purifiedby columnchromatography on silica gel using suitablesystems of solvents. For generating N-acetylatedvariants of AA-CAM, additional acetylation wasperformed by reacting the unprotected AA-CAMderivatives with the N-acetylsuccinimide. Purity andchemical structures of obtained compounds wereconfirmed by HPLC, LC-MS, and NMR spectroscopy(see Supplementary Methods).
With 1-ethyl-(3-(3-dimethylamino)propyl)-carbodiimide hydrochloride In dichloromethane at 20℃; for 1h; Cooling with ice;
7 Example 7
The NHS 1 eq was dissolved in dry dichloromethane, 1.2 eq of Boc-protected glycine was added, and EDCI 1.8 eq was added under ice-cooling conditions. After half an hour of ice-bathing reaction, the reaction was repeated for about 1 hour at room temperature, and quenched by adding water. The reaction mixture was extracted with methylene chloride.1.2 eq of this product was added to dry dichloromethane, then 1 eq of the final compound of Example 1 was added, and finally DIPEA 2 eq was added and stirred for about 2 hours.The reaction was detected by thin layer chromatography and found to be completely reacted. The reaction was quenched with water and the reaction mixture was extracted with dichloromethane.Combine the concentrated organic phase and directly input the next reaction without purification.The ethyl acetate solution of hydrochloric acid was added thereto,The reaction was monitored by thin layer chromatography until the starting material disappeared and the reaction solution was sparged.Extract with dichloromethane and wash with saturated sodium bicarbonate.The organic phases were combined and subjected to column chromatography to give the product.
With dicyclohexyl-carbodiimide In N,N-dimethyl-formamide at 20℃;
2.1 2-1.
The synthesis of the active ester of N-Boc protected amino acid was as follows. A linear amino acid with an amino at the terminal and length within 6 carbon atoms (one of glycine, 3-aminopropionic acid, 4-aminobutyric acid, 5-aminopentanoic acid, 6-aminocaproic acid) was saturated in 15 water at room temperature, and 16 di-tert-butyl carbonate dicarbonate (7 Boc-O-Boc) was added in a molar amount of twice that in in 17 THF to obtain a mixture. The mixture was stirred for a reaction for 2 hours at room temperature followed by an addition of 18 isopropanol to precipitate a large amount of the intermediate product, which was re-dissolved in a minimum amount of 6 DMF. 9 Diethyl ether was added to obtain a precipitate and the dissolution and precipitation were repeated twice to obtain 19 N-Boc protected amino acid. Finally, the 19 N-Boc protected amino acid was dissolved in 6 DMF, followed by an addition of 21 DCC and 22 NHS (a molar ratio 1:1:1), and stirred at room temperature for a reaction overnight. The precipitate was filtered off, and the supernatant was added with 9 diethyl ether to collect a precipitate, which was dissolved in a minimum amount of hot DMF, and precipitated by diethyl ether three times to obtain the active 23 ester of N-Boc protected amino acid.
With dicyclohexyl-carbodiimide In chloroform
With dicyclohexyl-carbodiimide In ethyl acetate at 20℃; for 24h;
General procedure for the synthesis of N4 amino acid-acylated nucleosides 4a-h
General procedure: Appropriate amino acid 1a-h (6.66 mmol) was dissolved in 24 mL of 1,4-dioxane and water mixture (1:1), 13.3 mL of 1 N NaOH (2 mol. eq.) was slowly added and the mixture was chilled in the freezer for 20 min. Di-tert-butyl dicarbonate (10 mmol) was added and the mixture was stirred for 3 hours at room temperature. The reaction was monitored by TLC analysis. After the reaction was completed, the solvent mixture was concentrated to 2-5 mL using a rotary evaporator and aqueous solution of citric acid (10%) was added until pH reached 2.0. The mixture was extracted with ethyl acetate. The organic phase was dried with anhydrous sodium sulphate and evaporated under reduced pressure to recover Boc-protected amino acid 2a-h. The protected amino acid 2a-h was dissolved in 30 mL of ethyl acetate, 7.33 mmol of N-hydroxysuccinimide and 7.33 mmol of N,N'-dicyclohexylcarbodiimide were added and the reaction mixture was stirred for 24 hours at room temperature. The formed white precipitate was filtered, ethyl acetate was removed under reduced pressure to obtain N-Boc protected and NHS-activated amino acid 3a-h. The protected-activated acid was dissolved in dimethylformamide (8-10 mL) and 2'-deoxycytidine (6.66 mmol) was added. The reaction mixture was stirred for 5 days at room temperature. After the reaction was completed, dimethylformamide was removed using a rotary evaporator and the residue was purified by column chromatography (silica gel, chloroform/methanol mixture, 100:0→85:15). The products were obtained in 34-85% yields. Synthesized nucleosides were characterized by NMR spectroscopy and HPLC-MS analysis.
1. Preparation of Boc-Gly-Pro-OSu. 15 g of L-proline was dissolved in 300 ml of 10% sodium bicarbonate. 150 ml of acetone and 27.2 g of Boc-Gly-OSu were added into the reaction mixture. The mixture was stirred for 5 h at RT. The mixture was washed with ether (1 x 300 ml). 500 ml of ethyl acetate was added into the aqueous layer. The pH of the mixture was adjusted to 2.4-2.5 with ice-cooled 3N HCl. The ethyl acetate layer was collected and washed with water (3 x 300 ml). The organic solution was dried over sodium sulfate. The solution was evaporated to dryness. 21 g of the residue (Boc-Gly-Pro-OH ) and 11 g of N-hydroxysuccinimide were dissolved in 300 ml of dichloromethlene. The mixture was cooled to 00C. 17 g of 1,3-dicyclohexylcarbodiimide was added into the reaction mixture. The mixture was stirred for 1 hour at 00C. The solid was removed by filtration. The dichloromethlene solution is washed with 5% sodium bicarbonate (I x 200 ml) and water ( 3 x 200 ml). The organic solution was dried over sodium sulfate. The solution was evaporated to dryness. Yield 23 g Boc- Gly-Pro-OSu.
With sodium hydrogencarbonate; In acetone; at 20℃; for 5h;
15 g of L-proline was dissolved in 300 ml of 10% sodium bicarbonate. 150 ml of acetone and 27.2 g of Boc-Gly-OSu were added into the reaction mixture. The mixture was stirred for 5 h at RT. The mixture was washed with ether (1×300 ml). 500 ml of ethyl acetate was added into the aqueous layer. The pH of the mixture was adjusted to 2.4-2.5 with ice-cooled 3 N HCl. The ethyl acetate layer was collected and washed with water (3×300 ml). The organic solution was dried over sodium sulfate. The solution was evaporated to dryness. 21 g of the residue (Boc-Gly-Pro-OH) and 11 g of N-hydroxysuccinimide were dissolved in 300 ml of dichloromethylene. The mixture was cooled to 0 C. 17 g of 1,3-dicyclohexylcarbodiimide was added into the reaction mixture. The mixture was stirred for 1 hour at 0 C. The solid was removed by filtration. The dichloromethylene solution was washed with 5% sodium bicarbonate (1×200 ml) and water (3×200 ml). The organic solution was dried over sodium sulfate. The solution was evaporated to dryness to yield 23 g Boc-Gly-Pro-OSu.
With N-ethyl-N,N-diisopropylamine; In dichloromethane; for 0.5h;
EXAMPLE 17 Synthesis of N-Boc-glycine-(3-carbobenzyloxyamido)propylamide. Di-isopropylethylamine (7.0 mL, 40 mmol) was added to a suspension of N-t-butyloxycarbonylglycine N-hydroxysuccinimide ester (5.56 g, 20 mmol) and <strong>[17400-34-9]N-carbobenzyloxy-1,3-diaminopropane hydrochloride</strong> (5.0 g, 20 mmol) in dichloromethane (50 ml). The solution became clear over several minutes. After 30 minutes, additional of N-t-butyloxycarbonylglycine N-hydroxysuccinimide ester (0.275 g, 1 mmol) was added. The solution was extracted with water, followed by saturated aqueous NaHCO3, then by 0.5 N HCl. The dichloromethane solution was filtered through a short column of Na2SO4 and evaporated in vacuo to obtain 4.1 g (56%) of N-BOC-Glycine-(3-carbobenzyloxyamido)propylamide. MS: m/e=366 (M+H+), 310 (M-C4H9+H+), 266 (M-BOC+H+).
With N-ethylbis(isobutyl)amine In dichloromethane at 0 - 20℃; for 17h;
40 Preparation 40
To a suspension of 4, 5-DIAMINO-L- (2- hydroxyethyl) pyrazole sulfuric acid salt (1.20 g) and N- [2- (TERT-BUTOXYCARBONYLAMINO) acetoxy] succinimide (1.35 g) in methylene chloride (20 ml) was added N- ethyldiisopropylamine (2.1 ml) under ice-cooling, and the mixture was stirred at room temperature for 17 hours. The reaction mixture was washed with water (40 ml), saturated aqueous sodium hydrogen carbonate solution (40 ml) and brine (40 ml). The organic layer was dried over anhydrous sodium sulfate, filtered and concentrated in vacuo. The oily residue was purified by column chromatography on silica gel eluting with 10% methanol/chloroform to give 5-AMINO-4- [2- (TERT- butoxycarbonylamino) acetyl] AMINO-1-(2- hydroxyethyl) pyrazole (1.20 g) as a solid. 1H-NMR (CDCL3) 8 1. 46 (9H, s), 3. 89-3. 90 (4H, m), 4. 00- 4.04 (2H, m), 4.26 (2H, br), 5.51 (1H, br), 7.17 (1H, s), 8.06 (1H, br)
tert-butyl 2-[(5-amino-1-methyl-1H-pyrazol-4-yl)amino]-2-oxoethylcarbamate[ No CAS ]
Yield
Reaction Conditions
Operation in experiment
With N-ethylbis(isobutyl)amine; In tetrahydrofuran; at 0 - 20℃;
To a suspension of 4, 5-DIAMINO-1-METHYLPYRAZOLE sulfuric acid salt (305 g) in tetrahydrofuran (3.05 L) was added tert-butyl 2-[(2, 5-DIOXO-1-PYRROLIDINYL) OXY]- 2-oxoethylcarbamate (415 g) under ice-cooling. To the mixture was added diisopropylethylamine (556 ml) dropwise at a temperature below 10C. The mixture was stirred at room temperature overnight. To the resulting solution were added brine and saturated aqueous sodium hydrogen carbonate solution, and the mixture was extracted with ethyl acetate (3.0 L). The aqueous layer was extracted with tetrahydrofuran/ethyl acetate=1/1 (3.0 L) twice. The organic layer was dried over anhydrous magnesium sulfate, filtered and concentrated in vacuo. The residue was triturated with diisopropyl ether (1.0 L) and dried in vacuo to give tert-butyl 2- [ (5-amino-l-methyl-lH-pyrazol-4-yl) amino] -2- oxoethylcarbamate (307 g). IR (KBR) 3440,3349, 1670,1631, 1525,1276, 1163,1074, 1014,860, 791 CRRI 1 1H-NMR (DMSO-d6) 8 1.39 (9H, s), 3.44 (3H, s), 3.64 (2H, d, J=5.9Hz), 4.91 (2H, brs), 6.97 (1H, t, J=5.9Hz), 7.15 (1H, s), 9.09 (1H, brs)
[] 50 mg (270 mumol) O-Phosphoryl-serine, 105 mg ethyl diisopropylamine and 110 mg (405 mumol) Boc-glycine-n-hyroxysuccinimide are reacted in methanol for 3 days at room temperature. The solvent is removed in vacuo and the remaining residue is digerated with methanol / dichloromethane and filtrated. The mother liquid is poured into diethyl ether. The precipitate is collected and purified by flash chromatography utilizing dichloromethane/methanol glacial acetic acid 10:1:0,1. The relevant fractions are collected, the solvent is removed in vacuo and the remaining residue is digerated with diethyl ether and dried in vacuo. 36 mg (39%) of the desired product are obtained. [TLC: acetonitrile/water/glacial acetic acid 5/1/0.2: Rf= 0.12]. (ESI-MS: m/e = 341 (M-H)-]
With N-ethyl-N,N-diisopropylamine In methanol at 20℃; for 3h;
A.6 Example A.6 Phosphate precursor:
[] 141 mg (1 mmol) O-Phosphoryl-ethanolamine, 356 mg ethyl diisopropylamine and 408 mg (1,5 mol) Boc-glycine-n-hyroxysuccinimide are reacted in methanol for 3 h at room temperature. The solvent is removed in vacuo and the remaining residue is digerated with methanol / dichloromethane and filtrated. The mother liquid is poured into diethyl ether. The precipitate is collected and purified by flash chromatography utilizing acetonitril/water/ glacial acetic acid 5:1:0,2. The relevant fractions are collected, the solvent is removed in vacuo and the remaining residue is digerated with diethyl ether and dried in vacuo. 170 mg (57%) of the desired product are obtained. [TLC: acetonitrile/water/glacial acetic acid 5/1/0,2: Rf = 0,2].
Step 1 Preparation of 1,1-dimethylethyl N-[2-[[[4-(5-methyl-3-phenylisoxazol-4-yl)phenyl]sulfonyl]amino]-2-oxoethyl]carbamate A mixture of 4-[5-methyl-3-(phenyl)isoxazol-4-yl]benzenesulfonamide (15.0 g, 47.7 mmol), N-t-boc-glycine N-hydroxysuccinimide ester (13.0 g, 47.7 mmol) and 1,8-diazobicyclo[4.3.0]undec-7-ene (14.5 g, 95.4 mmol) were mixed together in tetrahydrofuran for 1 hour at room temperature. Additional N-t-boc-glycine N-hydroxysuccinimide ester (1.3 g, 4.7 mmol) was added and the solution was stirred an additional 2 hours. The solvent was removed at reduced pressure and residue was taken up in ethyl acetate. The ethyl acetate was washed with 10% aqueous HCl, sat. aq. NaHCO3, dried over anhydrous Na2 SO4, filtered and concentrated in vacuo to afford the desired amide as a clear glassy solid (6.5 g, 75%): mp 160.2-162.0 C. 1 H NMR (CDCl3 /300 MHz) 8.04 (d, 2H, J=8.4 Hz), 7.44-7.33 (m, 5H), 7.28 (d, 2H, J=8.4 Hz), 5.24 (brs, 1H), 3.85 (m, 2H), 2.50 (s, 3H), 1.43 (s, 9H). FABLRMS m/z 472 (M+H). Anal. Calc'd for C23 H25 N3 O5 S.0.18 H2 O: C, 58.19; H, 5.38; N, 8.85. Found: C, 58.22; H, 5.73; N, 8.92.
R.160 Preparation of Boc-Gly-Leu--OH
REFERENCE EXAMPLE 160 Preparation of Boc-Gly-Leu--OH Using 2.65 g of H-Leu--OH and 5.0 g of Boc-Gly-OSu, the same procedure as in Reference Example 155 was repeated to obtain 3.71 g (yield: 69.9%) of the above objective compound having a melting point of 135°-137° C.
With triethylamine; In tetrahydrofuran; at 5 - 20℃; for 20h;
II.2c: Ethyl 3-(Boc-Glycinylamino)-3-phenylpropionate Boc-Glycine-hydroxysuccinimide ester (5 g) was slowly added at 5 C to a solution of II.2b (4.2 g) and triethylamine (5.1 ml) in 45 ml of THF. The solution was stirred at room temperature for 20 h, and the insoluble precipitate was removed by filtration and washed with THF. The collected solutions were concentrated, the residue was taken up in ethyl acetate and the mixture was washed with satd. NaHCO3 solution and water. After drying (MgSO4), it was concentrated and a viscous residue was obtained (yield: 6.3 g). 1H-NMR (CDCl3): 1.16 (t, 3 H), 1.45 (s, 9 H), 2.82 (dd, 1 H), 2.92 (dd, 1 H), 3.81 (m, 2 H), 4.06 (q, 2 H), 5.17 (m, 1 H), 5.43 (s, 1 H), 7.23-7.38 (m, 5 H).
With hydrogenchloride; sodium hydroxide; trifluoroacetic acid; In methanol; dichloromethane; chloroform; water;
a. N-Glycylthyroxine 380 mg of <strong>[51-48-9]L-Thyroxin</strong>e was dissolved in 2.0 mls of methanol on addition of 174.5 mul of 6N NaOH and sonication. 140 mg of N-t-BOC-Glycine-N-hydroxysuccinimide ester was added and the mixture vortexed. TLC indicated complete reaction within 10 min. 190 mul of 6N HCl and 5 mls of water were added and the mixture vortexed. The sticky precipitate was separated from the mixture with 3 extractions of 3 mls of chloroform. The organic fractions were dried with anhydrous sodium sulfate and the solvent stripped off, leaving 470 mg of an off-white solid. This was dissolved in 2 mls of methylene chloride and 2 mls of trifluoroacetic acid. After 10 min., the solvent was stripped off and the residue dissolved in 5 mls of methanol. The solvent was stripped off, leaving 500 mg of a light brown solid.
With sodium hydrogencarbonate; In 1,4-dioxane; water;
3,4-Dehydroproline 1 (0.011 g, 0.097 mmol) and sodium hydrogen carbonate (0.0082 g, 0.097 mmol) were dissolved in water (1 cm3). A solution of N-terf-butyloxycarbonylglycine N- hydroxysuccinimide 2 (0.024 g, 0.088 mmol) in dioxane (1 cm3) was added dropwise and stirred overnight. The reaction mixture was diluted with water, acidified with solid citric acid and extracted with dichloromethane (3 x). The combined organic layers were dried (MgSC^), filtered and the solvent removed to afford crude acid 3 (0.0242 g, ca. 100%) that was used without further purification. Acid 3 was shown to be a 75:25 trans :cis mixture of conformers by 1H NMR analysis (the ratio was estimated from the integration of the Proalpha protons at delta 5.21 and 5.13 of the major and minor conformers respectively): deltan (300 MHz; CDCl3; Me4Si) 1.411 [2.25H, s, C(CHs)3], 1.42 EPO <DP n="70"/>[6.75H, s, C(CHa)3], 3.83-4.08 (2Eta, m, GIyO-H2), 4.25-4.45 (2H, m, Prodelta-H2), 5.13* (0.25H, br d, J 2.8, Proalpha-H), 5.21 (0.75H, br d, /3.1, Proalpha-H), 5.55-5.80 (2H, br m, 0-H and N-H) and 5.86-6.02 (2H, m, Probeta-H and Progamma-H).
Production of the drug substance Gly-T3 ' HCl was performed as a one-pot reaction inside a 20-gallon glass lined reactor. To limit exposure of the workers to the potent material, the starting reagent T3 was prepared as a slurry in THF inside an EPO <DP n="44"/>isolator. The resulting slurry was then transferred to the 20-gallon reactor through a Teflon line. Subsequently, Boc-Gly-OSu was dissolved in THF, transferred to the 20-gallon reactor and then DIEA was dissolved in THF, and also transferred to the 20-gallon reactor. The suspension was stirred for 15 hours, at ambient temperature and became a clear solution. In process testing by HPLC (% AUC) of the collected sample after 15 hours showed a purity of 98.5 % for the desired compound, Boc- Gly-T3. The reaction was then quenched by adding water and the THF was removed by distillation. The solvent, TBME was then added and the batch was allowed to stir overnight at 5 to 10 0C. The solution was acidified by a 20 % aqueous Na2SO4ZNaHSO4 buffer solution. The product was extracted in TBME and the organic layers were combined in a 60 L Pope tank and dried with Na2SO4. The dried solution was then filtered through a 0.22 mum filter, charged into the 20- gallon reactor and stirred overnight at 15 0C. The TBME was removed by distillation, followed by IPAc chases (continuous feed), while the batch stirred overnight at 15 0C. The intermediate was Boc deprotected by pressurizing the 20- gallon reactor headspace with 30 psi HCl gas for 3 hours and 30 minutes. The obtained precipitate was filtered inside the isolator, washed with IPAc, and dried for 110 hours inside a vacuum oven at 35 0C. In process testing by GC showed the presence of 6.13 % residual IPAc. Therefore, the solids were hydrated (water displacement) for approximately 24 hours and dried until constant weight, in a vacuum oven for 50 hours at 35 0C. The material was packaged in glass amber bottles, which were put in polyethylene bags and stored at - 20 0C with desiccants.
The alcohol 5 (0.818 g, 4.36 mmol) and Boc-glycine N-hydroxysuccinimide ester (Boc- GIy- OSu) (Sigma-Aldrich, 1.835 g, 6.45 mmol) in dry pyridine (3.5 ml) were stirred at 55 - 65 0C for 1 h. The reaction mixture was diluted with toluene (30 ml) and dried in vacuo. The residue (3.28 g) was dissolved in ethyl acetate (40 ml), diluted with hexane (100 ml), filtered and washed with 10% NaCl (50 ml). The organic layer was dried over Na2SC^, filtered from the desiccant and dried in vacuo. The crude compound 6 (1.80 g) was purified by flash- chromatography on silica-gel, elution with mixtures ofhexane and ethyl acetate (5: 1 to 1 :1 by volume). Yield of pure compound 6: 1.41 g (94%). TLC of compound 6 (heptane - ethyl acetate, 2:3): one spot, Rf ca. 0.7. 1H NMR of compound 6 (CDCl3), δ, ppm: 1.42 (s, 9H), 3.02 (t, 7Hz, 2H), 3.88 (d, 6Hz, 2H), 4.38 (t, 7Hz, 2H), 4.98 (br., IH), 7.08 (m, IH), 7.60 - 7.66 (m, 2H), 8.45 (m, IH).
EXAMPLE 17 Synthesis of N-BOC-Glycine-(3-carbobenzyloxyamido)propylamide: Di-isopropylethylamine (7.0 mL, 40 mmol) was added to a suspension of N-t-butyloxycarbonylglycine N-hydroxysuccinimide ester (5.56 g, 20 mmol) and <strong>[17400-34-9]N-carbobenzyloxy-1,3-diaminopropane hydrochloride</strong> (5.0 g, 20 mmol) in dichloromethane (50 ml). The solution became clear over several minutes. After 30 minutes, additional of N-t-butyloxycarbonylglycine N-hydroxysuccinimide ester (0.275 g, 1 mmol) was added. The solution was extracted with water, followed by saturated aqueous NaHCO3, then by 0.5 N HCl. The dichloromethane solution was filtered through a short column of Na2SO4 and evaporated in vacuo to obtain 4.1 g (56%) of N-BOC-Glycine-(3-carbobenzyloxyamido)propylamide. MS: m/e=366 (M+H+), 310 (M-C4H9+H+), 266 (M-BOC+H+).
With 4-methyl-morpholine In N,N-dimethyl-formamide at 20℃;
2
To (rac)-mexiletine hydrochloride (3.08 g, 14.24 mmol) and 4-methylmorpholine (3.3 L, 3.03 g, 29.9 mmol) in dry DMF (60 mL) was added /V-f-butyloxycarbonyl-glycine succinimide (4.27 g, 15.67 mmol) and the solution stirred at room temperature overnight. Ethyl acetate (100 mL) was added and the solution quenched [NaCI: AcOH: H20; 0.45 g: 0.05 g: 180 mL] (100 mL) with stirring for 30 minutes. The organics54 were collected and quenched (100 mL) again for a further period of 30 minutes. After this time, the organics were collected and washed with 8% aqueous sodium bicarbonate (100 mL), 7% brine (100 mL), dried (MgS04) and concentrated to give the desired A/-f-butyloxycarbonyl-glycine-(rac)-mexiletine (4.92 g, 93 %), as a white solid.
With N-ethyl-N,N-diisopropylamine In dichloromethane at 15℃; for 3h;
3 Step 3: (S)-fert-Butyl 2-(2-carbamoylpyrrolidine-1-yl)-2-oxoethylcarbamate
Step 3: (S)-fert-Butyl 2-(2-carbamoylpyrrolidine-1-yl)-2-oxoethylcarbamate. A solution of the N-hydroxysuccinimidate obtained from step 2 (8.22g, 46.9mmol) in dichloromethane (100 mL) was cooled in a cooling bath to 15°C. After 15 min, a solution of (S)- pyrrolidine-2-carboxamide (5.1 g, 44.7 mmol, bought from commercial supplier Fluorochem) and 2.1 eq of Hunig's base in 50 m L of dichloromethane were added. After 3h, the resulting solution was washed with 1 N hydrochloric acid, with saturated sodium bicarbonate, brine, dried over sodium sulfate, filtrated and evaporated to dryness. The crude mixture was purified using flash chromatography with 5% methanol in DCM. Yield: 8.61 g, 71 % 1 H NMR (400 MHz, DMSO -cfe): δ 1 .38 (s, 9H), 1 .72-2.22 (m , 4H), 3.38-3.45 (m,1 H), 3.49-3.55 (m,1 H), 3,74 (d, J = 5.7 Hz, 2H), 4.18 (dd, J =8.8, 2.8 Hz, 1 H), 5.43 (br s, 1 H), 5.54 (br_s, 1 H) , 6.83 (br s, 1 H) MS (ESI) m/z 294.2 [M + Na]+.
71%
Stage #1: tert-butyl N-{2-[(2,5-dioxopyrrolidin-1-yl)oxy]-2-oxoethyl}carbamate In dichloromethane at 15℃; for 0.25h;
Stage #2: L-prolinamide In dichloromethane
3 Step 3: (S)-tert-Butyl 2-(2-carbamoylpyrrolidine-1-yl)-2-oxoethylcarbamate
A solution of the N-hydroxysuccinimidate obtained from step 2 (8.22 g, 46.9 mmol) in dichloromethane (100 mL) was cooled in a cooling bath to 15° C. After 15 min, a solution of (S)-pyrrolidine-2-carboxamide (5.1 g, 44.7 mmol, bought from commercial supplier Fluorochem) and 2.1 eq of Hünig's base in 50 mL of dichloromethane were added. After 3 h, the resulting solution was washed with 1N hydrochloric acid, with saturated sodium bicarbonate, brine, dried over sodium sulfate, filtrated and evaporated to dryness. The crude mixture was purified using flash chromatography with 5% methanol in DCM. Yield: 8.61 g, 71% 1H NMR (400 MHz, DMSO-d6): δ 1.38 (s, 9H), 1.72-2.22 (m, 4H), 3.38-3.45 (m, 1H), 3.49-3.55 (m, 1H), 3.74 (d, J=5.7 Hz, 2H), 4.18 (dd, J=8.8, 2.8 Hz, 1H), 5.43 (br s, 1H), 5.54 (br_s, 1H), 6.83 (br s, 1H) MS (ESI) m/z 294.2 [M+Na]+.
With triethylamine; In dimethyl sulfoxide; at 60℃; for 1h;
General procedure: Triethylamine (Et3N) (8 mmol) followed by BocGlyONSu or BocGly2ONSu (3 mmol) were added to a solution of diamine (1 mmol) in dimethylsulfoxide (DMSO; 5 mL). Reaction mixture was stirred till the disappearing of the starting diamine (1-24 h, TLC control) and the solvent was removed under a vacuum. Dry residue was suspended in methanol, filtered, dissolved in water, sedimented with methanol and dried in vacuum. BocGly-NH(H2)10NH-GlyBoc (22) was prepared according to Protocol 1 from 1,10-diaminodekane (1.2 g, 6.9 mmol) and BocGlyONSu (5.35 g, 20.9 mmol) in DMSO (20 mL). The mixture was stirred for 1 h at 60 , evaporated, and DMSO was removed in vacuum. The obtained white residue was suspended in MeOH and the precipitate was washed with MeOH and dried in vacuum. It was obtained 3.22 g of -derivative 22 as white powder (95%).
Stage #1: tert-butyl N-{2-[(2,5-dioxopyrrolidin-1-yl)oxy]-2-oxoethyl}carbamate With triethylamine In tetrahydrofuran at 20℃; for 0.5h;
Stage #2: L-arginine In tetrahydrofuran for 18h;
1.3
In the third step, weighed 12 g of BOC-glycine-OSu prepared in the second step, added to 200 mL of tetrahydrofuran solution, and then added 10 mL of triethylamine to the system, stirred at room temperature for 30 minutes, and then added 25 g of the compound of formula (3) to continue the reaction. After 18 hours, TLC monitored the progress of the reaction. After the reaction was completed, the mother liquor was extracted with (3 × 300 mL) ethyl acetate, and the aqueous phase was collected and passed.After adding ethanol several times, it is concentrated under reduced pressure to remove water. The residue is dissolved in anhydrous methanol, added to silica gel, and purified by column chromatography to obtain a snowy white foamy solid product (BOC-glycine-arginine). Pure product 24.65g, yield is 80%, TLC shows only one point;
In tetrahydrofuran; N,N-dimethyl-formamide at 20℃; for 2h;
5.1 5-1
1,7-N,N-dimethylamino-diethyltriamine was reacted with the active ester of N-Boc protected amino acid as follows. 1,7-N,N-dimethylamino-diethyltriamine was dissolved in DMF, and the THF saturated solution of the active ester of N-Boc protected amino acid prepared in step 2-1 of Embodiment 2 (a molar ratio of 1:1) was added, and the mixture was stirred at room temperature for a reaction for 2 hours. After the reaction, diethyl ether was added to precipitate. Finally, a process consisting of the dissolution with a minimum amount of DMF and the precipitation with diethyl ether was repeated four times to obtain 4-(N′-Boc-aminoacyl)-1,7-N,N-dimethylamino-diethyltriamine.
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