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CAS No. : | 23500-04-1 | MDL No. : | MFCD00236761 |
Formula : | C13H24N2O5 | Boiling Point : | - |
Linear Structure Formula : | - | InChI Key : | NZAMQYCOJSWUAO-JTQLQIEISA-N |
M.W : | 288.34 | Pubchem ID : | 7019610 |
Synonyms : |
|
Num. heavy atoms : | 20 |
Num. arom. heavy atoms : | 0 |
Fraction Csp3 : | 0.77 |
Num. rotatable bonds : | 11 |
Num. H-bond acceptors : | 5.0 |
Num. H-bond donors : | 3.0 |
Molar Refractivity : | 73.89 |
TPSA : | 104.73 Ų |
GI absorption : | High |
BBB permeant : | No |
P-gp substrate : | No |
CYP1A2 inhibitor : | No |
CYP2C19 inhibitor : | No |
CYP2C9 inhibitor : | No |
CYP2D6 inhibitor : | No |
CYP3A4 inhibitor : | No |
Log Kp (skin permeation) : | -7.39 cm/s |
Log Po/w (iLOGP) : | 2.04 |
Log Po/w (XLOGP3) : | 0.94 |
Log Po/w (WLOGP) : | 1.27 |
Log Po/w (MLOGP) : | 0.67 |
Log Po/w (SILICOS-IT) : | 0.8 |
Consensus Log Po/w : | 1.15 |
Lipinski : | 0.0 |
Ghose : | None |
Veber : | 1.0 |
Egan : | 0.0 |
Muegge : | 0.0 |
Bioavailability Score : | 0.55 |
Log S (ESOL) : | -1.49 |
Solubility : | 9.25 mg/ml ; 0.0321 mol/l |
Class : | Very soluble |
Log S (Ali) : | -2.73 |
Solubility : | 0.542 mg/ml ; 0.00188 mol/l |
Class : | Soluble |
Log S (SILICOS-IT) : | -2.25 |
Solubility : | 1.6 mg/ml ; 0.00556 mol/l |
Class : | Soluble |
PAINS : | 0.0 alert |
Brenk : | 0.0 alert |
Leadlikeness : | 1.0 |
Synthetic accessibility : | 2.92 |
Signal Word: | Warning | Class: | N/A |
Precautionary Statements: | P261-P305+P351+P338 | UN#: | N/A |
Hazard Statements: | H315-H319-H335 | Packing Group: | N/A |
GHS Pictogram: |
* 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.
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
87.7% | With potassium carbonate In tetrahydrofuran; water at 0 - 20℃; for 4 h; | Example A73 Preparation of Ar2-acetyl-L-lysyl-L-valyl-A^-carbamoyl-Ar-[4-([(2-[(3i?,51S',7i?,8i?)-8-hydroxy-7- {(l£,3£)-5-[(21S,,35',5i?,6i?)-5-[(2Z,41S)-4-hydroxypent-2-enoyl]amino}-3,6-dimethyltetrahydro- 2H-pyran-2-yl]-3-methylpenta-l,3-dien-l-yl}-l,6-dioxaspiro[2.5]oct-5- yl]acetyl}hydrazinyl)carbonyl]oxy}methyl)phenyl]-L-ornithinamide, acetate salt (B222). B227 Step 1. Synthesis of N2-acetyl-N6-(teri-butoxycarbonyl)-L-lysine (B223). To a mixture of N6- (fert-butoxycarbonyl)-L-lysine (22.5 g, 91.5 mmol, 1 eq.) and K2CO3 (63.1 g, 0.457 mol, 5 eq.) in tetrahydrofuran/water (200 mL/200 mL) at 0 °C was added acetyl chloride (8.62 g, 0.109 mol, 1.2 eq.), and the mixture was stirred at rt for 4 h. The mixture was concentrated in vacuo to remove the tetrahydrofuran, and the aqueous layer was adjusted to pH = 1 with 2 M HCI and extracted with EtOAc (100 mL) three times. The extract was washed with brine (100 mL), dried over sodium sulfate and concentrated in vacuo to afford B223 (23.1 g, 87.7 percent) as a yellow oil. Step 2. Synthesis of N2-acetyl-L-lysine hydrochloride salt (B224). To a solution of B223 (23.1 g, 0.080 mmol, 1 eq.) in ethyl acetate (400 mL) at 0 °C was added HC1 (g) in ethyl acetate (250 mL) under nitrogen. The mixture was stirred at rt for 4 h and filtered. The solid was washed with ethyl acetate and dried in vacuo to afford B224 (18.5 g, >100 percent) as a white solid which was used without further purification. Step 3. Synthesis of N2-acetyl-N6-[(9H-fluoren-9-ylmethoxy)carbonyl]-L-lysine (B225). To a mixture of B224 (8 g, 35.6 mmol, 1 eq.) and NaHC03 (5.99 g, 71.3 mmol, 2 eq.) in acetone/water (80 mL/80 mL) at 0 °C was added a solution of Fmoc-Cl (9.41 g, 36.3 mmol, 1.02 eq.) in acetone (80 mL), and the mixture was stirred at rt for 2 h. The mixture was adjusted to pH = 3-4 with 2 N HC1 and extracted with ethyl acetate (100 mL) three times. The extracts were washed with brine (100 mL), dried over sodium sulfate and concentrated in vacuo to give the crude product (7 g) as a yellow oil. To the crude product was added dichloromethane and fert-butylmethyl ether (100 mL), and the suspension was stirred for 30 min and then filtered. The filter cake was dried in vacuo to afford B225 (3.25 g, 22.2 percent) as a white solid. Step 4. Synthesis of N2-acetyl-N6-[(9H-fluoren-9-ylmethoxy)carbonyl]-L-lysyl-L-valyl-N5- carbamoyl-N-[4-(hydroxymethyl)phenyl]-L-ornithinamide (B226). To a mixture of B225 (1.04 g, 2.54 mmol, 1 eq.) in NN-dimethylformamide (20 mL) at 0 °C was added N-methylmorpholine (769 mg, 7.61 mmol, 3 eq.), l -ethyl-3-(3-dimethylaminopropyl) carbodiimide-HCl (632 mg, 3.30 mmol, 1.3 eq.), 1 -hydroxybenzotriazole hydrate (445 mg, 3.30 mmol, 1.3 eq.) and L-valyl-N5-carbamoyl-N-[4- (hydroxymethyl)phenyl]-L-ornithinamide (From WO04010957, 1.01 g, 2.66 mmol, 1.05 eq.) under nitrogen, and the mixture was stirred at rt for 2 h. the mixture was poured into teri-butylmethyl ether (300 mL) and filtered. The solid was washed with dichloromethane (50 mL) and water (50 mL) and dried in vacuo to afford B226 (1.87 g, 95.6percent) as a white solid. Step 5. Synthesis of N2-acetyl-N6-[(9H-fluoren-9-ylmethoxy)carbonyl]-L-lysyl-L-valyl-N5- carbamoyl-N-[4-( [(4-nitrophenoxy)carbonyl]oxy}methyl)phenyl]-L-ornithinamide (B227). To a mixture of B226 (1.87 g, 2.43 mmol, 1 eq.) and bis-(4-nitrophenyl)carbonate (2.21 g, 7.28 mmol, 3 eq.) in NN-dimethylformamide (30 mL) at 0 °C was added NN-diisopropylethylamine (313 mg, 2.43 mmol, 1 eq.) under nitrogen, and the mixture was stirred at rt overnight. The mixture was poured into teri-butylmethylether (50 mL) and filtered. The solid (1.95 g) was purified by prep HPLC to give B227 (580 mg, 25.7 percent) as a white solid. lU NMR (400Hz, DMSO-dg): 10.1 (s, 1 H), 8.29 (d, 2 H), 8.00 (d, 1 H), 7.86 (d, 1 H), 7.65 (d, 2 H), 7.64 (d, 1 H), 7.61 (m, 4 H), 7.40 (m, 2 H), 7.38 (m, 4 H), 7.30 (m, 3 H), 6.01 (br, 1 H), 5.21 (s, 2 H), 4.35 (br, 1 H), 4.27-4.15 (m, 5 H), 2.96 (m, 4 H), 1.98 (m, 1 H), 1.82 (s, 3 H), 1.65 (br, 3 H), 1.43-1.24 (m, 7 H), 0.83 (m, 6 H). Step 6. Synthesis ofN2-acetyl-N6-[(9H-fluoren-9-ylmethoxy)carbonyl]-L-lysyl-L-valyl-N5- carbamoyl-N-[4-( [(2-[(3R,5S,7R,8R)-8-hydroxy-7-{(l£',3£')-5-[(2S,3S,5R,6R)-5- [(2Z,4S)-4- hydroxypent-2-enoyl]amino}-3,6-dimethyltetrahydro-2H-pyran-2-yl]-3-methylpenta-l,3-dien-l -yl} - 1.6- dioxaspiro [2.5] oct-5-yl] acetyl} hydrazinyl)carbonyl] oxy } methyl)phenyl] -L-ornithinamide (B228) . To a solution of B209 (8.1 mg, 0.016 mmol, 1 eq.) in NN-dimethylformamide (0.4 mL) at rt was added 2,6-lutidine (7.5 \L, 0.064 mmol, 4 eq.), NN-diisopropylethylamine (11.3 \L, 0.064 mmol, 4 eq.) and 4-NN-dimethylamino pyridine (2 mg, 0.016 mmol, 1 eq.) followed by B227 (17.8 mg, 0.019 mmol, 1.2 eq.), and the reaction was stirred for 5 h. The reaction was purified by reverse phase chromatography (Method A) to give B228 as a white solid. Yield: 5.5 mg, 0.004 mmol, 26percent. LCMS (Protocol D): m/z 1306.1 [M+H]+, retention time = 0.81 minutes. Step 7. Synthesis of N2-acetyl-L-lysyl-L-valyl-N5-carbamoyl-N-[4-([(2- [(3R,5S,7R,8R)-8- hydroxy-7-{(l£',3£')-5-[(2S,3S,5R,6R)-5-[(2Z,4S)-4-hydroxypent-2-enoyl]amino}-3,6- dimethyltetrahydro-2H-pyran-2-yl]-3-methylpenta-l,3-dien-l -yl} -l,6-dioxaspiro[2.5]oct-5- yl]acetyl}hydrazinyl)carbonyl]oxy}methyl)phenyl]-L-ornithinamide, acetate salt (B222). The title compound was prepared in 79percent yield from 9.5 mg (0.007 mmol, 1.0 eq.) of B228 and 11.9 mg (0.14 mmol, 20.0 eq.) of piperidine using the procedure described for preparation of compound B47. LCMS (Protocol D): m/z 1084.1 [M+H]+, retention time = 0.58 minutes. lU NMR (500 MHz, DMSO-d6) δ 10.10 (s, 1 H), 8.22-8.12 (m, 1 H), 8.03 (d, J= 7.8 Hz, 1 H), 7.87-7.74 (m, 2 H), 7.64-7.53 (m, 2 H), 7.34-7.18 (m, 2 H), 6.31 (d, J= 15.9 Hz, 1 H), 6.09-6.01 (m, 1 H), 5.98 (d, J= 11.8 Hz, 1 H), 5.86 (dd, J= 11.8 and 7.1 Hz, 1 H), 5.66-5.56 (m, 1 H), 5.55-5.49 (m, 1 H), 5.44 (br s, 1 H), 5.23-4.91 (m, 3 H), 4.43-4.33 (m, 1 H), 4.30-4.21 (m, 2 H), 4.20-4.12 (m, 1 H), 3.69-3.59 (m, 1 H), 3.53-3.45 (m, 1 H), 3.07- 2.88 (m, 2 H), 2.76-2.71 (m, 1 H), 2.61-2.56 (m, 1 H), 2.35-2.14 (m, 4 H), 2.04-1.53 (m, 18 H), 1.52-1.18 (m, 10 H), 1.11 (d, J= 6.4 Hz, 3 H), 1.06 (d, J= 6.4 Hz, 3 H), 0.95 (d, J= 7.3 Hz, 3 H), 0.85 (d, J= 6.9 Hz, 3 H), 0.82 (d, J= 6.9 Hz, 3 H). |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
100% | With dicyclohexyl-carbodiimide; In ethyl acetate; at 0 - 20℃; | To a stirred solution of A/"-acetyl-A/£-Boc-(S)-lysine (1 .00g, 3.47 mmol) and A/-hydroxysuccinimide (0.44 g, 3.81 mmol) in ethyl acetate (25 mL) at 0 C was added Lambda/,Lambda/'-dicyclohexylcarbodi-imide (0.79 g, 3.81 mmol) and the mixture was stirred at this temperature for 2 h and then at room temperature overnight. The resulting suspension was filtered through Celite and the filtrate was concentrated to give N -acetyl - A -Boc-(S)-lysine A/-hydroxysuccinimide ester (1.44 g, 100 %), as a white solid which was used in the next step without further purification. |
1.49 mg | With dicyclohexyl-carbodiimide; In tetrahydrofuran; for 2h;Inert atmosphere; | Preparation of (S)-2,5-Dioxopyrrolidin-1-yl 2-acetamido-6-((tert-butoxycarbonyl)amino)hexanoate A solution of <strong>[23500-04-1](S)-2-acetamido-6-((tert-butoxycarbonyl)amino)hexanoic acid</strong> (1.00 g, 3.47 mmol) in tetrahydrofuran (20 mL) was treated with N-hydroxysuccinimide (439 mg, 3.81 mmol) and N,N'-dicyclohexylcarbodiimide (785 mg, 3.81 mmol) and stirred under a nitrogen atmosphere for 2 h. After this time, the reaction mixture was filtered to remove the solid dicyclohexylurea byproduct. The solid was washed with diethyl ether (100 mL), and the combined filtrate and washings were concentrated under reduced pressure. The residue was triturated with diethyl ether to provide (S)-2,5-dioxopyrrolidin-1-yl 2-acetamido-6-((tert-butoxycarbonyl)amino)hexanoate (1.49 mg) as a white solid: 1H NMR (300 MHz, CDCl3) 6.27 (m, 1H), 4.94 (m, 1H), 4.69 (m, 1H), 3.15-3.13 (m, 2H), 2.87 (s, 4H), 2.08 (s, 3H), 2.02-1.83 (m, 2H), 1.55-1.22 (m, 4H), 1.45 (s, 9H). |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
General procedure: 2-chlorotrityl chloride resin (0.05 mmol, 75 mg, 0.67 mmol/g) was swelled by stirring in CH2Cl2 (5 mL) at 25 C for 30 min. Next, a pre-mixed solution of Fmoc-Lys-OAllyl·HCl salt (0.5 mmol, 222.4 mg) and N,N?-diisopropylethylamine (DIPEA, 0.5 mmol, 88 muL) pre-dissolved in anhydrous CH2Cl2 (5 mL) was added to the resin and allowed to react at 25 C, 30 min. Unreacted reagents and solvent were removed by filtration and the resin washed with dimethylformamide (DMF, 15 x 3 mL) and CH2Cl2 (15 x 3 mL) followed by DMF (15 x 3 mL). Next, 20% piperidine in DMF (v/v, 5 mL) was added to the resin. The mixture was subjected to microwave irradiation while stirring at 60 C for 10 min. Unreacted reagents and solvent were removed by filtration and the resin washed with DMF (15 x 3 mL) and followed by CH2Cl2 (15 x 3 mL). Next, a premixed solution of Ac-Lys(Boc)-OH (0.5 mmol, 144 mg), N,N?-diisopropylcarbodiimide (DIC, 0.25 mmol, 27 muL) and hydroxybenzotriazole (HOBt, 0.25 mmol, 33.8 mg) dissolved in 90% CH2Cl2 in DMF (v/v, 5 mL) was added to the resin and the mixture was subjected to microwaveirradiation while stirring at 60 C for 10 min. Unreacted reagents and solvent were removed by filtration and the resin washed with DMF (15 x 3 mL) and CH2Cl2 (15mL) followed by CHCl3 (15 x 3 mL). The -OAllyl protecting group was removed by adding a pre-mixed solution of Pd0[P(Ph3)4] (5 mol %, 3 mg), phenylsilane (0.05 mmol, 7 muL) in 5 mL anhydrous CHCl3 and microwaved at 40 C for 30 min. This step was repeated. Excess reagents and solvent were drained and the resin was washed with CHCl3 (15 x 3 mL) followed by CH2Cl2 (15 x 3 mL). In the next step, a premixed solution of the appropriate amine (0.5 mmol), DIC (0.25 mmol, 27 muL) and HOBt (0.25 mmol, 33.8 mg) dissolved in 90% CH2Cl2 in DMF (v/v, 5 mL) was addedto the resin. The reaction mixture was subjected to microwave irradiation while stirring at 60 C for 10 min. Unreacted reagents and solvent were removed by filtration and the resin washed with CH2Cl2 (15 x 3 mL), DMF (15 x 3 mL) followed by CH2Cl2 (15 x 3 mL). The target was cleaved from the resin using 95% TFA in CH2Cl2 (v/v, 1.5 mL) at 25 C over 30 min. Excess TFA was removed by a stream of N2(g) to yield a colorless oil which was dissolved in MeOH (1 mL) and purified by HPLC (C18 column, H2O and CH3CN solvent). Target compounds were lyophilized in vacuo to give white powders (10-20 mg). |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
General procedure: 2-chlorotrityl chloride resin (0.05 mmol, 75 mg, 0.67 mmol/g) was swelled by stirring in CH2Cl2 (5 mL) at 25 C for 30 min. Next, a pre-mixed solution of Fmoc-Lys-OAllyl·HCl salt (0.5 mmol, 222.4 mg) and N,N?-diisopropylethylamine (DIPEA, 0.5 mmol, 88 muL) pre-dissolved in anhydrous CH2Cl2 (5 mL) was added to the resin and allowed to react at 25 C, 30 min. Unreacted reagents and solvent were removed by filtration and the resin washed with dimethylformamide (DMF, 15 x 3 mL) and CH2Cl2 (15 x 3 mL) followed by DMF (15 x 3 mL). Next, 20% piperidine in DMF (v/v, 5 mL) was added to the resin. The mixture was subjected to microwave irradiation while stirring at 60 C for 10 min. Unreacted reagents and solvent were removed by filtration and the resin washed with DMF (15 x 3 mL) and followed by CH2Cl2 (15 x 3 mL). Next, a premixed solution of Ac-Lys(Boc)-OH (0.5 mmol, 144 mg), N,N?-diisopropylcarbodiimide (DIC, 0.25 mmol, 27 muL) and hydroxybenzotriazole (HOBt, 0.25 mmol, 33.8 mg) dissolved in 90% CH2Cl2 in DMF (v/v, 5 mL) was added to the resin and the mixture was subjected to microwaveirradiation while stirring at 60 C for 10 min. Unreacted reagents and solvent were removed by filtration and the resin washed with DMF (15 x 3 mL) and CH2Cl2 (15mL) followed by CHCl3 (15 x 3 mL). The -OAllyl protecting group was removed by adding a pre-mixed solution of Pd0[P(Ph3)4] (5 mol %, 3 mg), phenylsilane (0.05 mmol, 7 muL) in 5 mL anhydrous CHCl3 and microwaved at 40 C for 30 min. This step was repeated. Excess reagents and solvent were drained and the resin was washed with CHCl3 (15 x 3 mL) followed by CH2Cl2 (15 x 3 mL). In the next step, a premixed solution of the appropriate amine (0.5 mmol), DIC (0.25 mmol, 27 muL) and HOBt (0.25 mmol, 33.8 mg) dissolved in 90% CH2Cl2 in DMF (v/v, 5 mL) was addedto the resin. The reaction mixture was subjected to microwave irradiation while stirring at 60 C for 10 min. Unreacted reagents and solvent were removed by filtration and the resin washed with CH2Cl2 (15 x 3 mL), DMF (15 x 3 mL) followed by CH2Cl2 (15 x 3 mL). The target was cleaved from the resin using 95% TFA in CH2Cl2 (v/v, 1.5 mL) at 25 C over 30 min. Excess TFA was removed by a stream of N2(g) to yield a colorless oil which was dissolved in MeOH (1 mL) and purified by HPLC (C18 column, H2O and CH3CN solvent). Target compounds were lyophilized in vacuo to give white powders (10-20 mg). |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
General procedure: 2-chlorotrityl chloride resin (0.05 mmol, 75 mg, 0.67 mmol/g) was swelled by stirring in CH2Cl2 (5 mL) at 25 C for 30 min. Next, a pre-mixed solution of Fmoc-Lys-OAllyl·HCl salt (0.5 mmol, 222.4 mg) and N,N?-diisopropylethylamine (DIPEA, 0.5 mmol, 88 muL) pre-dissolved in anhydrous CH2Cl2 (5 mL) was added to the resin and allowed to react at 25 C, 30 min. Unreacted reagents and solvent were removed by filtration and the resin washed with dimethylformamide (DMF, 15 x 3 mL) and CH2Cl2 (15 x 3 mL) followed by DMF (15 x 3 mL). Next, 20% piperidine in DMF (v/v, 5 mL) was added to the resin. The mixture was subjected to microwave irradiation while stirring at 60 C for 10 min. Unreacted reagents and solvent were removed by filtration and the resin washed with DMF (15 x 3 mL) and followed by CH2Cl2 (15 x 3 mL). Next, a premixed solution of Ac-Lys(Boc)-OH (0.5 mmol, 144 mg), N,N?-diisopropylcarbodiimide (DIC, 0.25 mmol, 27 muL) and hydroxybenzotriazole (HOBt, 0.25 mmol, 33.8 mg) dissolved in 90% CH2Cl2 in DMF (v/v, 5 mL) was added to the resin and the mixture was subjected to microwaveirradiation while stirring at 60 C for 10 min. Unreacted reagents and solvent were removed by filtration and the resin washed with DMF (15 x 3 mL) and CH2Cl2 (15mL) followed by CHCl3 (15 x 3 mL). The -OAllyl protecting group was removed by adding a pre-mixed solution of Pd0[P(Ph3)4] (5 mol %, 3 mg), phenylsilane (0.05 mmol, 7 muL) in 5 mL anhydrous CHCl3 and microwaved at 40 C for 30 min. This step was repeated. Excess reagents and solvent were drained and the resin was washed with CHCl3 (15 x 3 mL) followed by CH2Cl2 (15 x 3 mL). In the next step, a premixed solution of the appropriate amine (0.5 mmol), DIC (0.25 mmol, 27 muL) and HOBt (0.25 mmol, 33.8 mg) dissolved in 90% CH2Cl2 in DMF (v/v, 5 mL) was addedto the resin. The reaction mixture was subjected to microwave irradiation while stirring at 60 C for 10 min. Unreacted reagents and solvent were removed by filtration and the resin washed with CH2Cl2 (15 x 3 mL), DMF (15 x 3 mL) followed by CH2Cl2 (15 x 3 mL). The target was cleaved from the resin using 95% TFA in CH2Cl2 (v/v, 1.5 mL) at 25 C over 30 min. Excess TFA was removed by a stream of N2(g) to yield a colorless oil which was dissolved in MeOH (1 mL) and purified by HPLC (C18 column, H2O and CH3CN solvent). Target compounds were lyophilized in vacuo to give white powders (10-20 mg). |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
General procedure: 2-chlorotrityl chloride resin (0.05 mmol, 75 mg, 0.67 mmol/g) was swelled by stirring in CH2Cl2 (5 mL) at 25 C for 30 min. Next, a pre-mixed solution of Fmoc-Lys-OAllyl·HCl salt (0.5 mmol, 222.4 mg) and N,N?-diisopropylethylamine (DIPEA, 0.5 mmol, 88 muL) pre-dissolved in anhydrous CH2Cl2 (5 mL) was added to the resin and allowed to react at 25 C, 30 min. Unreacted reagents and solvent were removed by filtration and the resin washed with dimethylformamide (DMF, 15 x 3 mL) and CH2Cl2 (15 x 3 mL) followed by DMF (15 x 3 mL). Next, 20% piperidine in DMF (v/v, 5 mL) was added to the resin. The mixture was subjected to microwave irradiation while stirring at 60 C for 10 min. Unreacted reagents and solvent were removed by filtration and the resin washed with DMF (15 x 3 mL) and followed by CH2Cl2 (15 x 3 mL). Next, a premixed solution of Ac-Lys(Boc)-OH (0.5 mmol, 144 mg), N,N?-diisopropylcarbodiimide (DIC, 0.25 mmol, 27 muL) and hydroxybenzotriazole (HOBt, 0.25 mmol, 33.8 mg) dissolved in 90% CH2Cl2 in DMF (v/v, 5 mL) was added to the resin and the mixture was subjected to microwaveirradiation while stirring at 60 C for 10 min. Unreacted reagents and solvent were removed by filtration and the resin washed with DMF (15 x 3 mL) and CH2Cl2 (15mL) followed by CHCl3 (15 x 3 mL). The -OAllyl protecting group was removed by adding a pre-mixed solution of Pd0[P(Ph3)4] (5 mol %, 3 mg), phenylsilane (0.05 mmol, 7 muL) in 5 mL anhydrous CHCl3 and microwaved at 40 C for 30 min. This step was repeated. Excess reagents and solvent were drained and the resin was washed with CHCl3 (15 x 3 mL) followed by CH2Cl2 (15 x 3 mL). In the next step, a premixed solution of the appropriate amine (0.5 mmol), DIC (0.25 mmol, 27 muL) and HOBt (0.25 mmol, 33.8 mg) dissolved in 90% CH2Cl2 in DMF (v/v, 5 mL) was addedto the resin. The reaction mixture was subjected to microwave irradiation while stirring at 60 C for 10 min. Unreacted reagents and solvent were removed by filtration and the resin washed with CH2Cl2 (15 x 3 mL), DMF (15 x 3 mL) followed by CH2Cl2 (15 x 3 mL). The target was cleaved from the resin using 95% TFA in CH2Cl2 (v/v, 1.5 mL) at 25 C over 30 min. Excess TFA was removed by a stream of N2(g) to yield a colorless oil which was dissolved in MeOH (1 mL) and purified by HPLC (C18 column, H2O and CH3CN solvent). Target compounds were lyophilized in vacuo to give white powders (10-20 mg). |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
General procedure: 2-chlorotrityl chloride resin (0.05 mmol, 75 mg, 0.67 mmol/g) was swelled by stirring in CH2Cl2 (5 mL) at 25 C for 30 min. Next, a pre-mixed solution of Fmoc-Lys-OAllyl·HCl salt (0.5 mmol, 222.4 mg) and N,N?-diisopropylethylamine (DIPEA, 0.5mmol, 88 muL) pre-dissolved in anhydrous CH2Cl2 (5 mL) was added to the resin and allowed to react at 25 C, 30 min. Unreacted reagents and solvent were removed by filtration and the resin washed with DMF (15 x 3 mL) and CH2Cl2 (15 x 3 mL) followed by DMF (15 x 3 mL). Next, 20% piperidine in DMF (v/v, 5 mL) was added to the resin. The mixture was subjected to microwave irradiation while stirring at60C for 10 min. Unreacted reagents and solvent were removed by filtration and theresin washed with DMF (15 x 3 mL) and followed by CH2Cl2 (15 x 3 mL). Next, a premixed solution of Ac-Lys(Boc)-OH (0.5 mmol, 144 mg), DIC (0.25 mmol, 27 muL) and HOBt (0.25 mmol, 33.8 mg) dissolved in 90% CH2Cl2 in DMF (v/v, 5 mL) wasadded to the resin and the mixture was subjected to microwave irradiation while stirring at 60 C for 10 min. Unreacted reagents and solvent were removed by filtration and the resin washed with DMF (15 x 3 mL) and CH2Cl2 (15 mL) followed by CHCl3 (15 x 3 mL). The -OAllyl protecting group was removed by adding a premixed solution of Pd0[P(Ph3)4] (5 mol %, 3 mg), phenylsilane (0.05 mmol, 7 muL) in 5mL anhydrous CHCl3 and microwaved at 40C for 30 min. This step was repeated. Excess reagents and solvent were drained and the resin was washed with CHCl3 (15 x 3 mL) followed by CH2Cl2 (15 x 3 mL). In the next step, a pre-mixed solution of theappropriate diamine (0.5 mmol), DIC (0.25 mmol, 27 muL) and HOBt (0.25 mmol,33.8 mg) dissolved in 90% CH2Cl2 in DMF (v/v, 5 mL) was added to the resin. The reaction mixture was subjected to microwave irradiation while stirring at 60 C for 10 min. Unreacted reagents and solvent were removed by filtration and the resin washedwith CH2Cl2 (15 x 3 mL), DMF (15 x 3 mL) followed by CH2Cl2 (15 x 3 mL). Next, guanidinylation was conducted using a pre-mixed solution of N,N?-bis(tertbutoxycarbonyl)thiourea (0.5 mmol, 146 mg), DIPEA (0.5 mmol, 88 muL) and DMF (5 mL) was added to the resin and the reaction mixture subjected to microwave irradiation while stirring at 60 C for 10 min. Unreacted reagents were removed by filtration and the resin washed with DMF (15 x 3 mL) and CH2Cl2 (15 x 3 mL). The target was cleaved from the resin using 95% TFA in CH2Cl2 (v/v, 1.5 mL) at 25 C over 30 min. Excess TFA was removed by a stream of N2 (g) to yield a colorless oil which was dissolved in MeOH (1 mL) and purified by HPLC (C18 column, H2O and CH3CN solvent). Target compounds were lyophilized in vacuo to give white powders (10-20 mg). |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
General procedure: 2-chlorotrityl chloride resin (0.05 mmol, 75 mg, 0.67 mmol/g) was swelled by stirring in CH2Cl2 (5 mL) at 25 C for 30 min. Next, a pre-mixed solution of Fmoc-Lys-OAllyl·HCl salt (0.5 mmol, 222.4 mg) and N,N?-diisopropylethylamine (DIPEA, 0.5 mmol, 88 muL) pre-dissolved in anhydrous CH2Cl2 (5 mL) was added to the resin and allowed to react at 25 C, 30 min. Unreacted reagents and solvent were removed by filtration and the resin washed with dimethylformamide (DMF, 15 x 3 mL) and CH2Cl2 (15 x 3 mL) followed by DMF (15 x 3 mL). Next, 20% piperidine in DMF (v/v, 5 mL) was added to the resin. The mixture was subjected to microwave irradiation while stirring at 60 C for 10 min. Unreacted reagents and solvent were removed by filtration and the resin washed with DMF (15 x 3 mL) and followed by CH2Cl2 (15 x 3 mL). Next, a premixed solution of Ac-Lys(Boc)-OH (0.5 mmol, 144 mg), N,N?-diisopropylcarbodiimide (DIC, 0.25 mmol, 27 muL) and hydroxybenzotriazole (HOBt, 0.25 mmol, 33.8 mg) dissolved in 90% CH2Cl2 in DMF (v/v, 5 mL) was added to the resin and the mixture was subjected to microwaveirradiation while stirring at 60 C for 10 min. Unreacted reagents and solvent were removed by filtration and the resin washed with DMF (15 x 3 mL) and CH2Cl2 (15mL) followed by CHCl3 (15 x 3 mL). The -OAllyl protecting group was removed by adding a pre-mixed solution of Pd0[P(Ph3)4] (5 mol %, 3 mg), phenylsilane (0.05 mmol, 7 muL) in 5 mL anhydrous CHCl3 and microwaved at 40 C for 30 min. This step was repeated. Excess reagents and solvent were drained and the resin was washed with CHCl3 (15 x 3 mL) followed by CH2Cl2 (15 x 3 mL). In the next step, a premixed solution of the appropriate amine (0.5 mmol), DIC (0.25 mmol, 27 muL) and HOBt (0.25 mmol, 33.8 mg) dissolved in 90% CH2Cl2 in DMF (v/v, 5 mL) was addedto the resin. The reaction mixture was subjected to microwave irradiation while stirring at 60 C for 10 min. Unreacted reagents and solvent were removed by filtration and the resin washed with CH2Cl2 (15 x 3 mL), DMF (15 x 3 mL) followed by CH2Cl2 (15 x 3 mL). The target was cleaved from the resin using 95% TFA in CH2Cl2 (v/v, 1.5 mL) at 25 C over 30 min. Excess TFA was removed by a stream of N2(g) to yield a colorless oil which was dissolved in MeOH (1 mL) and purified by HPLC (C18 column, H2O and CH3CN solvent). Target compounds were lyophilized in vacuo to give white powders (10-20 mg). |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
General procedure: 2-chlorotrityl chloride resin (0.05 mmol, 75 mg, 0.67 mmol/g) was swelled by stirring in CH2Cl2 (5 mL) at 25 C for 30 min. Next, a pre-mixed solution of Fmoc-Lys-OAllyl·HCl salt (0.5 mmol, 222.4 mg) and N,N?-diisopropylethylamine (DIPEA, 0.5mmol, 88 muL) pre-dissolved in anhydrous CH2Cl2 (5 mL) was added to the resin and allowed to react at 25 C, 30 min. Unreacted reagents and solvent were removed by filtration and the resin washed with DMF (15 x 3 mL) and CH2Cl2 (15 x 3 mL) followed by DMF (15 x 3 mL). Next, 20% piperidine in DMF (v/v, 5 mL) was added to the resin. The mixture was subjected to microwave irradiation while stirring at60C for 10 min. Unreacted reagents and solvent were removed by filtration and theresin washed with DMF (15 x 3 mL) and followed by CH2Cl2 (15 x 3 mL). Next, a premixed solution of Ac-Lys(Boc)-OH (0.5 mmol, 144 mg), DIC (0.25 mmol, 27 muL) and HOBt (0.25 mmol, 33.8 mg) dissolved in 90% CH2Cl2 in DMF (v/v, 5 mL) wasadded to the resin and the mixture was subjected to microwave irradiation while stirring at 60 C for 10 min. Unreacted reagents and solvent were removed by filtration and the resin washed with DMF (15 x 3 mL) and CH2Cl2 (15 mL) followed by CHCl3 (15 x 3 mL). The -OAllyl protecting group was removed by adding a premixed solution of Pd0[P(Ph3)4] (5 mol %, 3 mg), phenylsilane (0.05 mmol, 7 muL) in 5mL anhydrous CHCl3 and microwaved at 40C for 30 min. This step was repeated. Excess reagents and solvent were drained and the resin was washed with CHCl3 (15 x 3 mL) followed by CH2Cl2 (15 x 3 mL). In the next step, a pre-mixed solution of theappropriate diamine (0.5 mmol), DIC (0.25 mmol, 27 muL) and HOBt (0.25 mmol,33.8 mg) dissolved in 90% CH2Cl2 in DMF (v/v, 5 mL) was added to the resin. The reaction mixture was subjected to microwave irradiation while stirring at 60 C for 10 min. Unreacted reagents and solvent were removed by filtration and the resin washedwith CH2Cl2 (15 x 3 mL), DMF (15 x 3 mL) followed by CH2Cl2 (15 x 3 mL). Next, guanidinylation was conducted using a pre-mixed solution of N,N?-bis(tertbutoxycarbonyl)thiourea (0.5 mmol, 146 mg), DIPEA (0.5 mmol, 88 muL) and DMF (5 mL) was added to the resin and the reaction mixture subjected to microwave irradiation while stirring at 60 C for 10 min. Unreacted reagents were removed by filtration and the resin washed with DMF (15 x 3 mL) and CH2Cl2 (15 x 3 mL). The target was cleaved from the resin using 95% TFA in CH2Cl2 (v/v, 1.5 mL) at 25 C over 30 min. Excess TFA was removed by a stream of N2 (g) to yield a colorless oil which was dissolved in MeOH (1 mL) and purified by HPLC (C18 column, H2O and CH3CN solvent). Target compounds were lyophilized in vacuo to give white powders (10-20 mg). |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
General procedure: 2-chlorotrityl chloride resin (0.05 mmol, 75 mg, 0.67 mmol/g) was swelled by stirring in CH2Cl2 (5 mL) at 25 C for 30 min. Next, a pre-mixed solution of Fmoc-Lys-OAllyl·HCl salt (0.5 mmol, 222.4 mg) and N,N?-diisopropylethylamine (DIPEA, 0.5mmol, 88 muL) pre-dissolved in anhydrous CH2Cl2 (5 mL) was added to the resin and allowed to react at 25 C, 30 min. Unreacted reagents and solvent were removed by filtration and the resin washed with DMF (15 x 3 mL) and CH2Cl2 (15 x 3 mL) followed by DMF (15 x 3 mL). Next, 20% piperidine in DMF (v/v, 5 mL) was added to the resin. The mixture was subjected to microwave irradiation while stirring at60C for 10 min. Unreacted reagents and solvent were removed by filtration and theresin washed with DMF (15 x 3 mL) and followed by CH2Cl2 (15 x 3 mL). Next, a premixed solution of Ac-Lys(Boc)-OH (0.5 mmol, 144 mg), DIC (0.25 mmol, 27 muL) and HOBt (0.25 mmol, 33.8 mg) dissolved in 90% CH2Cl2 in DMF (v/v, 5 mL) wasadded to the resin and the mixture was subjected to microwave irradiation while stirring at 60 C for 10 min. Unreacted reagents and solvent were removed by filtration and the resin washed with DMF (15 x 3 mL) and CH2Cl2 (15 mL) followed by CHCl3 (15 x 3 mL). The -OAllyl protecting group was removed by adding a premixed solution of Pd0[P(Ph3)4] (5 mol %, 3 mg), phenylsilane (0.05 mmol, 7 muL) in 5mL anhydrous CHCl3 and microwaved at 40C for 30 min. This step was repeated. Excess reagents and solvent were drained and the resin was washed with CHCl3 (15 x 3 mL) followed by CH2Cl2 (15 x 3 mL). In the next step, a pre-mixed solution of theappropriate diamine (0.5 mmol), DIC (0.25 mmol, 27 muL) and HOBt (0.25 mmol,33.8 mg) dissolved in 90% CH2Cl2 in DMF (v/v, 5 mL) was added to the resin. The reaction mixture was subjected to microwave irradiation while stirring at 60 C for 10 min. Unreacted reagents and solvent were removed by filtration and the resin washedwith CH2Cl2 (15 x 3 mL), DMF (15 x 3 mL) followed by CH2Cl2 (15 x 3 mL). Next, guanidinylation was conducted using a pre-mixed solution of N,N?-bis(tertbutoxycarbonyl)thiourea (0.5 mmol, 146 mg), DIPEA (0.5 mmol, 88 muL) and DMF (5 mL) was added to the resin and the reaction mixture subjected to microwave irradiation while stirring at 60 C for 10 min. Unreacted reagents were removed by filtration and the resin washed with DMF (15 x 3 mL) and CH2Cl2 (15 x 3 mL). The target was cleaved from the resin using 95% TFA in CH2Cl2 (v/v, 1.5 mL) at 25 C over 30 min. Excess TFA was removed by a stream of N2 (g) to yield a colorless oil which was dissolved in MeOH (1 mL) and purified by HPLC (C18 column, H2O and CH3CN solvent). Target compounds were lyophilized in vacuo to give white powders (10-20 mg). |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
General procedure: 2-chlorotrityl chloride resin (0.05 mmol, 75 mg, 0.67 mmol/g) was swelled by stirring in CH2Cl2 (5 mL) at 25 C for 30 min. Next, a pre-mixed solution of Fmoc-Lys-OAllyl·HCl salt (0.5 mmol, 222.4 mg) and N,N?-diisopropylethylamine (DIPEA, 0.5mmol, 88 muL) pre-dissolved in anhydrous CH2Cl2 (5 mL) was added to the resin and allowed to react at 25 C, 30 min. Unreacted reagents and solvent were removed by filtration and the resin washed with DMF (15 x 3 mL) and CH2Cl2 (15 x 3 mL) followed by DMF (15 x 3 mL). Next, 20% piperidine in DMF (v/v, 5 mL) was added to the resin. The mixture was subjected to microwave irradiation while stirring at60C for 10 min. Unreacted reagents and solvent were removed by filtration and theresin washed with DMF (15 x 3 mL) and followed by CH2Cl2 (15 x 3 mL). Next, a premixed solution of Ac-Lys(Boc)-OH (0.5 mmol, 144 mg), DIC (0.25 mmol, 27 muL) and HOBt (0.25 mmol, 33.8 mg) dissolved in 90% CH2Cl2 in DMF (v/v, 5 mL) wasadded to the resin and the mixture was subjected to microwave irradiation while stirring at 60 C for 10 min. Unreacted reagents and solvent were removed by filtration and the resin washed with DMF (15 x 3 mL) and CH2Cl2 (15 mL) followed by CHCl3 (15 x 3 mL). The -OAllyl protecting group was removed by adding a premixed solution of Pd0[P(Ph3)4] (5 mol %, 3 mg), phenylsilane (0.05 mmol, 7 muL) in 5mL anhydrous CHCl3 and microwaved at 40C for 30 min. This step was repeated. Excess reagents and solvent were drained and the resin was washed with CHCl3 (15 x 3 mL) followed by CH2Cl2 (15 x 3 mL). In the next step, a pre-mixed solution of theappropriate diamine (0.5 mmol), DIC (0.25 mmol, 27 muL) and HOBt (0.25 mmol,33.8 mg) dissolved in 90% CH2Cl2 in DMF (v/v, 5 mL) was added to the resin. The reaction mixture was subjected to microwave irradiation while stirring at 60 C for 10 min. Unreacted reagents and solvent were removed by filtration and the resin washedwith CH2Cl2 (15 x 3 mL), DMF (15 x 3 mL) followed by CH2Cl2 (15 x 3 mL). Next, guanidinylation was conducted using a pre-mixed solution of N,N?-bis(tertbutoxycarbonyl)thiourea (0.5 mmol, 146 mg), DIPEA (0.5 mmol, 88 muL) and DMF (5 mL) was added to the resin and the reaction mixture subjected to microwave irradiation while stirring at 60 C for 10 min. Unreacted reagents were removed by filtration and the resin washed with DMF (15 x 3 mL) and CH2Cl2 (15 x 3 mL). The target was cleaved from the resin using 95% TFA in CH2Cl2 (v/v, 1.5 mL) at 25 C over 30 min. Excess TFA was removed by a stream of N2 (g) to yield a colorless oil which was dissolved in MeOH (1 mL) and purified by HPLC (C18 column, H2O and CH3CN solvent). Target compounds were lyophilized in vacuo to give white powders (10-20 mg). |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
General procedure: 2-chlorotrityl chloride resin (0.05 mmol, 75 mg, 0.67 mmol/g) was swelled by stirring in CH2Cl2 (5 mL) at 25 C for 30 min. Next, a pre-mixed solution of Fmoc-Lys-OAllyl·HCl salt (0.5 mmol, 222.4 mg) and N,N?-diisopropylethylamine (DIPEA, 0.5mmol, 88 muL) pre-dissolved in anhydrous CH2Cl2 (5 mL) was added to the resin and allowed to react at 25 C, 30 min. Unreacted reagents and solvent were removed by filtration and the resin washed with DMF (15 x 3 mL) and CH2Cl2 (15 x 3 mL) followed by DMF (15 x 3 mL). Next, 20% piperidine in DMF (v/v, 5 mL) was added to the resin. The mixture was subjected to microwave irradiation while stirring at60C for 10 min. Unreacted reagents and solvent were removed by filtration and theresin washed with DMF (15 x 3 mL) and followed by CH2Cl2 (15 x 3 mL). Next, a premixed solution of Ac-Lys(Boc)-OH (0.5 mmol, 144 mg), DIC (0.25 mmol, 27 muL) and HOBt (0.25 mmol, 33.8 mg) dissolved in 90% CH2Cl2 in DMF (v/v, 5 mL) wasadded to the resin and the mixture was subjected to microwave irradiation while stirring at 60 C for 10 min. Unreacted reagents and solvent were removed by filtration and the resin washed with DMF (15 x 3 mL) and CH2Cl2 (15 mL) followed by CHCl3 (15 x 3 mL). The -OAllyl protecting group was removed by adding a premixed solution of Pd0[P(Ph3)4] (5 mol %, 3 mg), phenylsilane (0.05 mmol, 7 muL) in 5mL anhydrous CHCl3 and microwaved at 40C for 30 min. This step was repeated. Excess reagents and solvent were drained and the resin was washed with CHCl3 (15 x 3 mL) followed by CH2Cl2 (15 x 3 mL). In the next step, a pre-mixed solution of theappropriate diamine (0.5 mmol), DIC (0.25 mmol, 27 muL) and HOBt (0.25 mmol,33.8 mg) dissolved in 90% CH2Cl2 in DMF (v/v, 5 mL) was added to the resin. The reaction mixture was subjected to microwave irradiation while stirring at 60 C for 10 min. Unreacted reagents and solvent were removed by filtration and the resin washedwith CH2Cl2 (15 x 3 mL), DMF (15 x 3 mL) followed by CH2Cl2 (15 x 3 mL). Next, guanidinylation was conducted using a pre-mixed solution of N,N?-bis(tertbutoxycarbonyl)thiourea (0.5 mmol, 146 mg), DIPEA (0.5 mmol, 88 muL) and DMF (5 mL) was added to the resin and the reaction mixture subjected to microwave irradiation while stirring at 60 C for 10 min. Unreacted reagents were removed by filtration and the resin washed with DMF (15 x 3 mL) and CH2Cl2 (15 x 3 mL). The target was cleaved from the resin using 95% TFA in CH2Cl2 (v/v, 1.5 mL) at 25 C over 30 min. Excess TFA was removed by a stream of N2 (g) to yield a colorless oil which was dissolved in MeOH (1 mL) and purified by HPLC (C18 column, H2O and CH3CN solvent). Target compounds were lyophilized in vacuo to give white powders (10-20 mg). |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
General procedure: 2-chlorotrityl chloride resin (0.05 mmol, 75 mg, 0.67 mmol/g) was swelled by stirring in CH2Cl2 (5 mL) at 25 C for 30 min. Next, a pre-mixed solution of Fmoc-Lys-OAllyl·HCl salt (0.5 mmol, 222.4 mg) and N,N?-diisopropylethylamine (DIPEA, 0.5mmol, 88 muL) pre-dissolved in anhydrous CH2Cl2 (5 mL) was added to the resin and allowed to react at 25 C, 30 min. Unreacted reagents and solvent were removed by filtration and the resin washed with DMF (15 x 3 mL) and CH2Cl2 (15 x 3 mL) followed by DMF (15 x 3 mL). Next, 20% piperidine in DMF (v/v, 5 mL) was added to the resin. The mixture was subjected to microwave irradiation while stirring at60C for 10 min. Unreacted reagents and solvent were removed by filtration and theresin washed with DMF (15 x 3 mL) and followed by CH2Cl2 (15 x 3 mL). Next, a premixed solution of Ac-Lys(Boc)-OH (0.5 mmol, 144 mg), DIC (0.25 mmol, 27 muL) and HOBt (0.25 mmol, 33.8 mg) dissolved in 90% CH2Cl2 in DMF (v/v, 5 mL) wasadded to the resin and the mixture was subjected to microwave irradiation while stirring at 60 C for 10 min. Unreacted reagents and solvent were removed by filtration and the resin washed with DMF (15 x 3 mL) and CH2Cl2 (15 mL) followed by CHCl3 (15 x 3 mL). The -OAllyl protecting group was removed by adding a premixed solution of Pd0[P(Ph3)4] (5 mol %, 3 mg), phenylsilane (0.05 mmol, 7 muL) in 5mL anhydrous CHCl3 and microwaved at 40C for 30 min. This step was repeated. Excess reagents and solvent were drained and the resin was washed with CHCl3 (15 x 3 mL) followed by CH2Cl2 (15 x 3 mL). In the next step, a pre-mixed solution of theappropriate diamine (0.5 mmol), DIC (0.25 mmol, 27 muL) and HOBt (0.25 mmol,33.8 mg) dissolved in 90% CH2Cl2 in DMF (v/v, 5 mL) was added to the resin. The reaction mixture was subjected to microwave irradiation while stirring at 60 C for 10 min. Unreacted reagents and solvent were removed by filtration and the resin washedwith CH2Cl2 (15 x 3 mL), DMF (15 x 3 mL) followed by CH2Cl2 (15 x 3 mL). Next, guanidinylation was conducted using a pre-mixed solution of N,N?-bis(tertbutoxycarbonyl)thiourea (0.5 mmol, 146 mg), DIPEA (0.5 mmol, 88 muL) and DMF (5 mL) was added to the resin and the reaction mixture subjected to microwave irradiation while stirring at 60 C for 10 min. Unreacted reagents were removed by filtration and the resin washed with DMF (15 x 3 mL) and CH2Cl2 (15 x 3 mL). The target was cleaved from the resin using 95% TFA in CH2Cl2 (v/v, 1.5 mL) at 25 C over 30 min. Excess TFA was removed by a stream of N2 (g) to yield a colorless oil which was dissolved in MeOH (1 mL) and purified by HPLC (C18 column, H2O and CH3CN solvent). Target compounds were lyophilized in vacuo to give white powders (10-20 mg). |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
General procedure: 2-chlorotrityl chloride resin (0.05 mmol, 75 mg, 0.67 mmol/g) was swelled by stirring in CH2Cl2 (5 mL) at 25 C for 30 min. Next, a pre-mixed solution of Fmoc-Lys-OAllyl·HCl salt (0.5 mmol, 222.4 mg) and N,N?-diisopropylethylamine (DIPEA, 0.5mmol, 88 muL) pre-dissolved in anhydrous CH2Cl2 (5 mL) was added to the resin and allowed to react at 25 C, 30 min. Unreacted reagents and solvent were removed by filtration and the resin washed with DMF (15 x 3 mL) and CH2Cl2 (15 x 3 mL) followed by DMF (15 x 3 mL). Next, 20% piperidine in DMF (v/v, 5 mL) was added to the resin. The mixture was subjected to microwave irradiation while stirring at60C for 10 min. Unreacted reagents and solvent were removed by filtration and theresin washed with DMF (15 x 3 mL) and followed by CH2Cl2 (15 x 3 mL). Next, a premixed solution of Ac-Lys(Boc)-OH (0.5 mmol, 144 mg), DIC (0.25 mmol, 27 muL) and HOBt (0.25 mmol, 33.8 mg) dissolved in 90% CH2Cl2 in DMF (v/v, 5 mL) wasadded to the resin and the mixture was subjected to microwave irradiation while stirring at 60 C for 10 min. Unreacted reagents and solvent were removed by filtration and the resin washed with DMF (15 x 3 mL) and CH2Cl2 (15 mL) followed by CHCl3 (15 x 3 mL). The -OAllyl protecting group was removed by adding a premixed solution of Pd0[P(Ph3)4] (5 mol %, 3 mg), phenylsilane (0.05 mmol, 7 muL) in 5mL anhydrous CHCl3 and microwaved at 40C for 30 min. This step was repeated. Excess reagents and solvent were drained and the resin was washed with CHCl3 (15 x 3 mL) followed by CH2Cl2 (15 x 3 mL). In the next step, a pre-mixed solution of theappropriate diamine (0.5 mmol), DIC (0.25 mmol, 27 muL) and HOBt (0.25 mmol,33.8 mg) dissolved in 90% CH2Cl2 in DMF (v/v, 5 mL) was added to the resin. The reaction mixture was subjected to microwave irradiation while stirring at 60 C for 10 min. Unreacted reagents and solvent were removed by filtration and the resin washedwith CH2Cl2 (15 x 3 mL), DMF (15 x 3 mL) followed by CH2Cl2 (15 x 3 mL). Next, guanidinylation was conducted using a pre-mixed solution of N,N?-bis(tertbutoxycarbonyl)thiourea (0.5 mmol, 146 mg), DIPEA (0.5 mmol, 88 muL) and DMF (5 mL) was added to the resin and the reaction mixture subjected to microwave irradiation while stirring at 60 C for 10 min. Unreacted reagents were removed by filtration and the resin washed with DMF (15 x 3 mL) and CH2Cl2 (15 x 3 mL). The target was cleaved from the resin using 95% TFA in CH2Cl2 (v/v, 1.5 mL) at 25 C over 30 min. Excess TFA was removed by a stream of N2 (g) to yield a colorless oil which was dissolved in MeOH (1 mL) and purified by HPLC (C18 column, H2O and CH3CN solvent). Target compounds were lyophilized in vacuo to give white powders (10-20 mg). |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
100% | With hydrogenchloride; In ethyl acetate; at 0 - 20℃; for 4h;Inert atmosphere; | Example A73 Preparation of Ar2-acetyl-L-lysyl-L-valyl-A^-carbamoyl-Ar-[4-([(2-[(3i?,51S',7i?,8i?)-8-hydroxy-7- {(l£,3£)-5-[(21S,,35',5i?,6i?)-5-[(2Z,41S)-4-hydroxypent-2-enoyl]amino}-3,6-dimethyltetrahydro- 2H-pyran-2-yl]-3-methylpenta-l,3-dien-l-yl}-l,6-dioxaspiro[2.5]oct-5- yl]acetyl}hydrazinyl)carbonyl]oxy}methyl)phenyl]-L-ornithinamide, acetate salt (B222). B227 Step 1. Synthesis of N2-acetyl-N6-(teri-butoxycarbonyl)-L-lysine (B223). To a mixture of N6- (fert-butoxycarbonyl)-L-lysine (22.5 g, 91.5 mmol, 1 eq.) and K2CO3 (63.1 g, 0.457 mol, 5 eq.) in tetrahydrofuran/water (200 mL/200 mL) at 0 C was added acetyl chloride (8.62 g, 0.109 mol, 1.2 eq.), and the mixture was stirred at rt for 4 h. The mixture was concentrated in vacuo to remove the tetrahydrofuran, and the aqueous layer was adjusted to pH = 1 with 2 M HCI and extracted with EtOAc (100 mL) three times. The extract was washed with brine (100 mL), dried over sodium sulfate and concentrated in vacuo to afford B223 (23.1 g, 87.7 %) as a yellow oil. Step 2. Synthesis of N2-acetyl-L-lysine hydrochloride salt (B224). To a solution of B223 (23.1 g, 0.080 mmol, 1 eq.) in ethyl acetate (400 mL) at 0 C was added HC1 (g) in ethyl acetate (250 mL) under nitrogen. The mixture was stirred at rt for 4 h and filtered. The solid was washed with ethyl acetate and dried in vacuo to afford B224 (18.5 g, >100 %) as a white solid which was used without further purification. Step 3. Synthesis of N2-acetyl-N6-[(9H-fluoren-9-ylmethoxy)carbonyl]-L-lysine (B225). To a mixture of B224 (8 g, 35.6 mmol, 1 eq.) and NaHC03 (5.99 g, 71.3 mmol, 2 eq.) in acetone/water (80 mL/80 mL) at 0 C was added a solution of Fmoc-Cl (9.41 g, 36.3 mmol, 1.02 eq.) in acetone (80 mL), and the mixture was stirred at rt for 2 h. The mixture was adjusted to pH = 3-4 with 2 N HC1 and extracted with ethyl acetate (100 mL) three times. The extracts were washed with brine (100 mL), dried over sodium sulfate and concentrated in vacuo to give the crude product (7 g) as a yellow oil. To the crude product was added dichloromethane and fert-butylmethyl ether (100 mL), and the suspension was stirred for 30 min and then filtered. The filter cake was dried in vacuo to afford B225 (3.25 g, 22.2 %) as a white solid. Step 4. Synthesis of N2-acetyl-N6-[(9H-fluoren-9-ylmethoxy)carbonyl]-L-lysyl-L-valyl-N5- carbamoyl-N-[4-(hydroxymethyl)phenyl]-L-ornithinamide (B226). To a mixture of B225 (1.04 g, 2.54 mmol, 1 eq.) in NN-dimethylformamide (20 mL) at 0 C was added N-methylmorpholine (769 mg, 7.61 mmol, 3 eq.), l -ethyl-3-(3-dimethylaminopropyl) carbodiimide-HCl (632 mg, 3.30 mmol, 1.3 eq.), 1 -hydroxybenzotriazole hydrate (445 mg, 3.30 mmol, 1.3 eq.) and L-valyl-N5-carbamoyl-N-[4- (hydroxymethyl)phenyl]-L-ornithinamide (From WO04010957, 1.01 g, 2.66 mmol, 1.05 eq.) under nitrogen, and the mixture was stirred at rt for 2 h. the mixture was poured into teri-butylmethyl ether (300 mL) and filtered. The solid was washed with dichloromethane (50 mL) and water (50 mL) and dried in vacuo to afford B226 (1.87 g, 95.6%) as a white solid. Step 5. Synthesis of N2-acetyl-N6-[(9H-fluoren-9-ylmethoxy)carbonyl]-L-lysyl-L-valyl-N5- carbamoyl-N-[4-( [(4-nitrophenoxy)carbonyl]oxy}methyl)phenyl]-L-ornithinamide (B227). To a mixture of B226 (1.87 g, 2.43 mmol, 1 eq.) and bis-(4-nitrophenyl)carbonate (2.21 g, 7.28 mmol, 3 eq.) in NN-dimethylformamide (30 mL) at 0 C was added NN-diisopropylethylamine (313 mg, 2.43 mmol, 1 eq.) under nitrogen, and the mixture was stirred at rt overnight. The mixture was poured into teri-butylmethylether (50 mL) and filtered. The solid (1.95 g) was purified by prep HPLC to give B227 (580 mg, 25.7 %) as a white solid. lU NMR (400Hz, DMSO-dg): 10.1 (s, 1 H), 8.29 (d, 2 H), 8.00 (d, 1 H), 7.86 (d, 1 H), 7.65 (d, 2 H), 7.64 (d, 1 H), 7.61 (m, 4 H), 7.40 (m, 2 H), 7.38 (m, 4 H), 7.30 (m, 3 H), 6.01 (br, 1 H), 5.21 (s, 2 H), 4.35 (br, 1 H), 4.27-4.15 (m, 5 H), 2.96 (m, 4 H), 1.98 (m, 1 H), 1.82 (s, 3 H), 1.65 (br, 3 H), 1.43-1.24 (m, 7 H), 0.83 (m, 6 H). Step 6. Synthesis ofN2-acetyl-N6-[(9H-fluoren-9-ylmethoxy)carbonyl]-L-lysyl-L-valyl-N5- carbamoyl-N-[4-( [(2-[(3R,5S,7R,8R)-8-hydroxy-7-{(l£',3£')-5-[(2S,3S,5R,6R)-5- [(2Z,4S)-4- hydroxypent-2-enoyl]amino}-3,6-dimethyltetrahydro-2H-pyran-2-yl]-3-methylpenta-l,3-dien-l -yl} - 1.6- dioxaspiro [2.5] oct-5-yl] acetyl} hydrazinyl)carbonyl] oxy } methyl)phenyl] -L-ornithinamide (B228) . To a solution of B209 (8.1 mg, 0.016 mmol, 1 eq.) in NN-dimethylformamide (0.4 mL) at rt was added 2,6-lutidine (7.5 L, 0.064 mmol, 4 eq.), NN-diisopropylethylamine (11.3 L, 0.064 mmol, 4 eq.) and 4-NN-dimethylamino pyridine (2 mg, 0.016 mmol, 1 eq.) followed by B227 (17.8 mg, 0.019 mmol, 1.2 eq.), and the reaction was stirred for 5 h. The reaction was purified by reverse phase chromatography (Method A) to give B228 as a white solid. Yield: 5.5 mg, 0.004 mmol, 26%. LCMS (Protocol D): m/z 1306.1 [M+H]+, retention time = 0.81 minutes. Step 7. Synthesis of N2-acetyl-L-lysyl-L-valyl... |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
87.7% | With potassium carbonate; In tetrahydrofuran; water; at 0 - 20℃; for 4h; | Example A73 Preparation of Ar2-acetyl-L-lysyl-L-valyl-A^-carbamoyl-Ar-[4-([(2-[(3i?,51S',7i?,8i?)-8-hydroxy-7- {(l£,3£)-5-[(21S,,35',5i?,6i?)-5-[(2Z,41S)-4-hydroxypent-2-enoyl]amino}-3,6-dimethyltetrahydro- 2H-pyran-2-yl]-3-methylpenta-l,3-dien-l-yl}-l,6-dioxaspiro[2.5]oct-5- yl]acetyl}hydrazinyl)carbonyl]oxy}methyl)phenyl]-L-ornithinamide, acetate salt (B222). B227 Step 1. Synthesis of N2-acetyl-N6-(teri-butoxycarbonyl)-L-lysine (B223). To a mixture of N6- (fert-butoxycarbonyl)-L-lysine (22.5 g, 91.5 mmol, 1 eq.) and K2CO3 (63.1 g, 0.457 mol, 5 eq.) in tetrahydrofuran/water (200 mL/200 mL) at 0 C was added acetyl chloride (8.62 g, 0.109 mol, 1.2 eq.), and the mixture was stirred at rt for 4 h. The mixture was concentrated in vacuo to remove the tetrahydrofuran, and the aqueous layer was adjusted to pH = 1 with 2 M HCI and extracted with EtOAc (100 mL) three times. The extract was washed with brine (100 mL), dried over sodium sulfate and concentrated in vacuo to afford B223 (23.1 g, 87.7 %) as a yellow oil. Step 2. Synthesis of N2-acetyl-L-lysine hydrochloride salt (B224). To a solution of B223 (23.1 g, 0.080 mmol, 1 eq.) in ethyl acetate (400 mL) at 0 C was added HC1 (g) in ethyl acetate (250 mL) under nitrogen. The mixture was stirred at rt for 4 h and filtered. The solid was washed with ethyl acetate and dried in vacuo to afford B224 (18.5 g, >100 %) as a white solid which was used without further purification. Step 3. Synthesis of N2-acetyl-N6-[(9H-fluoren-9-ylmethoxy)carbonyl]-L-lysine (B225). To a mixture of B224 (8 g, 35.6 mmol, 1 eq.) and NaHC03 (5.99 g, 71.3 mmol, 2 eq.) in acetone/water (80 mL/80 mL) at 0 C was added a solution of Fmoc-Cl (9.41 g, 36.3 mmol, 1.02 eq.) in acetone (80 mL), and the mixture was stirred at rt for 2 h. The mixture was adjusted to pH = 3-4 with 2 N HC1 and extracted with ethyl acetate (100 mL) three times. The extracts were washed with brine (100 mL), dried over sodium sulfate and concentrated in vacuo to give the crude product (7 g) as a yellow oil. To the crude product was added dichloromethane and fert-butylmethyl ether (100 mL), and the suspension was stirred for 30 min and then filtered. The filter cake was dried in vacuo to afford B225 (3.25 g, 22.2 %) as a white solid. Step 4. Synthesis of N2-acetyl-N6-[(9H-fluoren-9-ylmethoxy)carbonyl]-L-lysyl-L-valyl-N5- carbamoyl-N-[4-(hydroxymethyl)phenyl]-L-ornithinamide (B226). To a mixture of B225 (1.04 g, 2.54 mmol, 1 eq.) in NN-dimethylformamide (20 mL) at 0 C was added N-methylmorpholine (769 mg, 7.61 mmol, 3 eq.), l -ethyl-3-(3-dimethylaminopropyl) carbodiimide-HCl (632 mg, 3.30 mmol, 1.3 eq.), 1 -hydroxybenzotriazole hydrate (445 mg, 3.30 mmol, 1.3 eq.) and L-valyl-N5-carbamoyl-N-[4- (hydroxymethyl)phenyl]-L-ornithinamide (From WO04010957, 1.01 g, 2.66 mmol, 1.05 eq.) under nitrogen, and the mixture was stirred at rt for 2 h. the mixture was poured into teri-butylmethyl ether (300 mL) and filtered. The solid was washed with dichloromethane (50 mL) and water (50 mL) and dried in vacuo to afford B226 (1.87 g, 95.6%) as a white solid. Step 5. Synthesis of N2-acetyl-N6-[(9H-fluoren-9-ylmethoxy)carbonyl]-L-lysyl-L-valyl-N5- carbamoyl-N-[4-( [(4-nitrophenoxy)carbonyl]oxy}methyl)phenyl]-L-ornithinamide (B227). To a mixture of B226 (1.87 g, 2.43 mmol, 1 eq.) and bis-(4-nitrophenyl)carbonate (2.21 g, 7.28 mmol, 3 eq.) in NN-dimethylformamide (30 mL) at 0 C was added NN-diisopropylethylamine (313 mg, 2.43 mmol, 1 eq.) under nitrogen, and the mixture was stirred at rt overnight. The mixture was poured into teri-butylmethylether (50 mL) and filtered. The solid (1.95 g) was purified by prep HPLC to give B227 (580 mg, 25.7 %) as a white solid. lU NMR (400Hz, DMSO-dg): 10.1 (s, 1 H), 8.29 (d, 2 H), 8.00 (d, 1 H), 7.86 (d, 1 H), 7.65 (d, 2 H), 7.64 (d, 1 H), 7.61 (m, 4 H), 7.40 (m, 2 H), 7.38 (m, 4 H), 7.30 (m, 3 H), 6.01 (br, 1 H), 5.21 (s, 2 H), 4.35 (br, 1 H), 4.27-4.15 (m, 5 H), 2.96 (m, 4 H), 1.98 (m, 1 H), 1.82 (s, 3 H), 1.65 (br, 3 H), 1.43-1.24 (m, 7 H), 0.83 (m, 6 H). Step 6. Synthesis ofN2-acetyl-N6-[(9H-fluoren-9-ylmethoxy)carbonyl]-L-lysyl-L-valyl-N5- carbamoyl-N-[4-( [(2-[(3R,5S,7R,8R)-8-hydroxy-7-{(l£',3£')-5-[(2S,3S,5R,6R)-5- [(2Z,4S)-4- hydroxypent-2-enoyl]amino}-3,6-dimethyltetrahydro-2H-pyran-2-yl]-3-methylpenta-l,3-dien-l -yl} - 1.6- dioxaspiro [2.5] oct-5-yl] acetyl} hydrazinyl)carbonyl] oxy } methyl)phenyl] -L-ornithinamide (B228) . To a solution of B209 (8.1 mg, 0.016 mmol, 1 eq.) in NN-dimethylformamide (0.4 mL) at rt was added 2,6-lutidine (7.5 L, 0.064 mmol, 4 eq.), NN-diisopropylethylamine (11.3 L, 0.064 mmol, 4 eq.) and 4-NN-dimethylamino pyridine (2 mg, 0.016 mmol, 1 eq.) followed by B227 (17.8 mg, 0.019 mmol, 1.2 eq.), and the reaction was stirred for 5 h. The reaction was purified by reverse phase chromatography (Method A) to give B228 as a white solid. Yield: 5.5 mg, 0.004 mmol, 26%. LCMS (Protocol D): m/z 1306.1 [M+H]+, retention time = 0.81 minutes. Step 7. Synthesis of N2-acetyl-L-lysyl-L-valyl... |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
92.8% | With benzotriazol-1-ol; O-(1H-benzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate; In N,N-dimethyl-formamide; for 0.333333h;Inert atmosphere; | To a 200 mL round bottom flask containing N-acetyl Lysine free acid (4.993 g, 17.32 mmol) were added HOBt (2.128 g, 15.75 mmol), HBTU (6.57 g, 17.32 mmol), compound 2 (6.18 g, 15.75 mmol) and DMF (40 mL). The solution was cooled via ice bath. DIEA (10 mL, 57.41 mmol) was added by syringe under Ar while stirred. The ice bath was removed after 20 min and the reaction was determined to be complete by LCMS. The reaction mixture was poured into water (300 mL) and extracted with ethyl acetate (2 x 100 mL). The organic layers were combined and washed with 1M NaHSO4 (3 x 100 mL), sat NaHCO3 (3 x 100 mL) and brine (2 x 100 ml). The organic layer was dried over Na2SO4, filtered and concentrated. The product was purified by silica gel column chromatography (4-5% MeOH/DCM, Rf=0.25) to obtain 7.19 g (92.8%). MWcal: 491.30; MWfd: 492.3 [M+H+]. 1H NMR (300 MHz, DMSO-d6) delta 1.05-1.65 (m, 22H), 1.82 (s, 3H), 2.34 (t, J=7.42 Hz, 2H), 2.86 (q, J=6.66 Hz, 2H), 2.93-3.10 (m, 2H), 4.06-4.21 (m, 1H), 5.08 (s, 2H), 6.75 (br t, J=5.50 Hz, 1H), 7.27-7.44 (m, 5H), 7.80-7.97 (m, 2H); 13C NMR (75 MHz, DMSO-d6) delta (ppm) 22.5, 22.7, 24.1, 25.7, 28.2, 28.6, 29.2, 31.9, 33.4, 38.2, 52.5, 65.3, 77.3, 127.9, 127.9, 128.4, 136.3, 155.5, 169.0, 171.5, 172.7 |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
The title compound was synthesized with classical methods of peptide chemistry starting with coupling of N2-acetyl-N6-(tert-butoxycarbonyl)-L-lysine with benzyl (2-aminoethyl)carbamate hydrochloride (1:1) with HATU and N,N-diisopropylethylamine and subsequent removal of the Z-protecting group by hydrogenation in DCM/methanol 1:1 over 10% palladiumlactivated charcoal under normal pressure. LC-MS (Method 1): R1 = 0.43 mm; MS (ESIpos): mz = 331 (M+H)+. | ||
The titel compound was obtained by coupling of N2-acetyl-N6-(tert-butoxycarbonyl)-L-lysinebenzyl (2-aminoethyl)carbamate hydrochloride (1:1) in DMF in the presence of HATIJ and N,Ndiisopropylethylamine and subsequent deprotection of the benzyloxycarbonyl group by hydrognenation in dichloromethane/methanol 1:1 in presence of 10% Pd/C at room temperature for 1 h.LC-MS (Methode 1): R1 = 0.43 mm; MS (ESIpos): m/z = 331 (M+H)t | ||
N2-Acetyl-N-(2-aminoethyl)-N6-(tert-butoxycarbonyl)-L-lysinamide The title compound was prepared by conventional methods of peptide chemistry by HATU coupling of commercially available N2-acetyl-N6-(tert-butoxycarbonyl)-L-lysine with benzyl (2-aminoethyl)carbamate hydrochloride (1:1) in the presence of N,N-diisopropylethylamine and subsequent detachment of the Z protecting group by hydrogenation in DCM/methanol 1:1 over 10% palladium on activated carbon. LC-MS (Method 1): Rt=0.43 min; MS (ESIpos): m/z=331 (M+H)+. |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
With pyridine; 1-ethyl-(3-(3-dimethylamino)propyl)-carbodiimide hydrochloride; In N,N-dimethyl-formamide; at 0 - 20℃;Inert atmosphere; | The compound, N,N?-Bis(alpha-acetyl-Lysine(epsilon-succinyl))-rhodamine 110 ((Ac-K(Succ.)2-R110) was prepared as follows. Pyridine (1 ml) and 1-ethyl-(3-dimethylaminopropyl)-carbodiimide hydrochloride (1.4 mmol, 0.268 g, Chem-Impex-International, Cat. 00050) was added to a solution of Rhodamine Green 560 chloride (rhodamine 110, 0.5 mmol, 0.183 g) and Ac-Lys(Boc)-OH (1.05 mmol, 0.303 g, Bachem Cat. E1040) in DMF (1 ml) at 0 C. under an argon atmosphere. The mixture was stirred at room temperature overnight. All volatility was evaporated by vacuum. The residue was purified by flash chromatography (silica gel; 5% to 15% MeOH in CH2Cl2) to yield a yellowish, semisolid impure intermediate (0.24 g, 28% yield). The intermediate (0.24 g, 0.27 mmol) was dissolved in CH2Cl2 (1.5 ml). Trifluoroacetic acid (1.5 ml) was added to this solution at 0 C. The mixture was stirred at room temperature for 1 hour. All the solvents were evaporated by rotary evaporator to give 0.20 g of the deprotected (deBOC) compound. Succinic anhydride (0.075 g, 0.075 mmol) was added to a suspension of the deBOC compound (0.10 g, 0.15 mmol) in DMF (1 ml). The resulting mixture was stirred overnight under an argon atmosphere at room temperature. All volatility was evaporated and the mixture was purified by flash chromatography (10% to 30% MeOH in CH2Cl2) to yield a yellow, solid mixture. The mixture was purified again by reverse phase chromatography (C18 silica gel, 100% H2O to 10% H2O in MeOH to 100% MeOH) to afford 0.022 g of pure compound: C44H50N6O13, FW: 870.90. The NMR spectrum was consistent with the structure depicted as Compound 2, N,N?-Bis(alpha-acetyl-Lysine(epsilon-succinyl))-rhodamine 110 ((Ac-K(Succ.))2-R110). |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
To a stirred solution of Ac-Lys(Boc)-OH (1.0 g, 3.5 mmol) in MeCN (40 mL) was added K2CO3 (1.4 g, 10.5 mmol). The reaction mixture was stirred for 10 min at room temperature. A solution of 5-bromoacetyl-2-hydroxybenzoic acid methyl ester (0.96 g, 3.5 mmol) in MeCN (10 mL) was added dropwise to the reaction mixture. The reaction mixture was stirred under ambient conditions. The progress of the reaction was monitored by TLC. After completion of the reaction, the reaction mixture was filtered and water (50 mL) and EtOAc (40 mL) were added to the reaction mixture. EtOAc layer was separated and aqueous layer was re-extracted with EtOAc (20 mL x 2). The EtOAc fractions were combined, dried over anhydrous MgSO4, filtered, and the supernatant concentrated under reduced pressure to afford the crude methyl 5-(2-((N2-acetyl-N6-(tert-butoxycarbonyl)lysyl)oxy)acetyl)-2-hydroxybenzoate as a yellowish white solid. This crude product was used in the next step without any purification. Crude methyl 5-(2-((N2-acetyl-N6-(tert-butoxycarbonyl)lysyl)oxy)acetyl)-2-hydroxybenzoate was dissolved in TFA/DCM (20 mL) and resulted solution was stirred under ambient conditions overnight. After completion of the reaction (Boc group removal) confirmed by TLC, the reaction mixture was concentrated under reduced pressure. The residue was dissolved in water and EtOAc (40 mL, 1:1). The water layer was separated, frozen, and then lyophilized to afford Methyl 5-(2-((acetyllysyl)oxy)acetyl)-2-hydroxybenzoate as a light pink colored sticky gummy solid (1.1 g, 85%). 1H NMR (400 MHz, D2O), acetic acid as an internal standard, delta (ppm): 8.36 (s, 1H), 8.17 - 7.71 (m, 1H), 7.04 (d, J = 8.8 Hz, 1H), 5.80 - 5.16 (m, 2H), 4.53 (dd, J = 8.9, 5.2 Hz, 1H), 3.94 (s, 3H), 2.99 (t, J = 7.6 Hz, 2H), 2.02 (s, 3H), 2.01 - 1.94 (m, 1H), 1.91 - 1.77 (m, 1H), 1.76 - 1.64 (m, 2H), 1.56 - 1.43 (m, 2H). 13C NMR (126 MHz, D2O) delta (ppm): 192.77, 173.86, 172.72, 169.17, 163.94, 162.35, 134.53, 131.20, 124.82, 117.68, 112.50, 66.60, 52.43, 51.94, 38.67, 29.54, 25.71, 21.47.HRMS (M + H) calculated for C18H25N2O7 380.16, found 381.1643. | ||
To a stirred solution of Ac-Lys(Boc)-OH (1.0 g, 3.5 mmol) in MeCN (40 mL) was added K2C03 (1.4 g, 10.5 mmol). The reaction mixture was stirred for 10 minutes at room temperature. A solution of 5-bromoacetyl-2-hydroxybenzoic acid methyl ester (0.96 g, 3.5 mmol) in MeCN (10 mL) was added dropwise to the reaction mixture. The reaction mixture was stirred under ambient conditions. The progress of the reaction was monitored by TLC. After completion of the reaction, the reaction mixture was filtered and water (50 mL) and EtOAc (40 mL) were added to the reaction mixture. EtOAc layer was separated and aqueous layer was re-extracted with EtOAc (20 mL x 2). The EtOAc fractions were combined, dried over anhydrous MgS04, filtered, and the supernatant concentrated under reduced pressure to afford the crude methyl 5-(2-((N2-acetyl-N6-(tert- butoxycarbonyl)lysyl)oxy)acetyl)-2-hydroxybenzoate as a yellowish white solid. This crude product was used in the next step without any purification. Crude methyl 5-(2-((N2-acetyl-N6- (tert-butoxycarbonyl)lysyl)oxy)acetyl)-2-hydroxybenzoate was dissolved in TFA/DCM (20 mL) and the resulted solution was stirred under ambient conditions overnight. After completion of the reaction (Boc group removal) confirmed by TLC, the reaction mixture was concentrated under reduced pressure. The residue was dissolved in water and EtOAc (40 mL, 1 : 1). The water layer was separated, frozen, and then lyophilized to afford Methyl 5-(2- ((acetyllysyl)oxy)acetyl)-2-hydroxybenzoate as a light pink colored solid (1.1 g, 85%). 1H NMR (400 MHz, D20), acetic acid as an internal standard, delta (ppm): 8.36 (s, 1H), 8.17 - 7.71 (m, 1H), 7.04 (d, J= 8.8 Hz, 1H), 5.80 - 5.16 (m, 2H), 4.53 (dd, J= 8.9, 5.2 Hz, 1H), 3.94 (s, 3H), 2.99 (t, J = 7.6 Hz, 2H), 2.02 (s, 3H), 2.01 - 1.94 (m, 1H), 1.91 - 1.77 (m, 1H), 1.76 - 1.64 (m, 2H), 1.56 - 1.43 (m, 2H). 13C NMR (126 MHz, D20) delta (ppm): 192.77, 173.86, 172.72, 169.17, 163.94, 162.35, 134.53, 131.20, 124.82, 117.68, 112.50, 66.60, 52.43, 51.94, 38.67, 29.54, 25.71, 21.47.HRMS (M + H) calculated for Ci8H25N207 380.16, found 381.1643. |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
Ac-Lys(Boc)-OH (1, 0.80 g, 2.77 mmol) and HATU (1.26 g, 3.33 mmol) were dissolved in anhydrous DMF(30 mL). H-Lys(Boc)-OtBu (2, 0.94 g, 2.77 mmol) was added together with DIPEA (1.68 mL, 9.70 mmol) tothe solution. The reaction was stirred for 5 h. the solvent was then removed in vacuo. Crude oil wasdissolved in DCM, water was added and the aqueous layer was extracted with DCM (3 x 20 mL). Theorganic layers were combined, washed with brine, dried over NaSO4 and concentrated in vacuo. Thecrude product was dissolved in TFA/DCM 1/1 (10 mL) and stirred for 5 hours. The solvent was removedunder gentle nitrogen flow and the product (3) was redissolved in water and DCM. The aqueous layerwas extracted with DCM (3 x 20 mL), the organic layers were combined, washed with brine, dried overNaSO4 and concentrated in vacuo. Half of the crude product (3) was redissolved in DMF/H2O (20 mL) andK2CO3 (1.30 g, 9.66 mmol) was added together with tert-butyl bromoacetate (4, 917 muL, 6.21 mmol). Thetemperature was brought to 60 C and the reaction was stirred for 30 minutes. The temperature wasbrought down to room temperature and acetic acid 5 % (20 mL) was mixed into the solution. DCM wasadded and the aqueous layer was extracted with DCM (3 x 20 mL). The organic layers were combined,washed with brine, dried over NaSO4, and concentrated in vacuo. The crude product was purified viasilica gel chromatography (hexane/EtOAc/MeOH 50/50/0 to 0/80/20) to achieve 5 as an off white solid(0.45 g, 0.58 mmol, 42 % yield over three steps). |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
97% | With N-ethyl-N,N-diisopropylamine; HATU; In N,N-dimethyl-formamide; at 20℃; for 0.5h; | To a solution of 217 (481 mg, 1.27 mmol) in 10 mL of DMF, 233 (366 mg, 1.27 mmol), HATU (660 mg, 1.65 mmol) and Hunig's base (0.302 mL, 1.6 mmol) where added. The reaction was allowed to stir at room temperature for 30 minutes. The reaction was diluted with ethyl acetate which caused to solids to crash out. This slurry was allowed to stir for 30 minutes. The solids were collected by filtration, rinsed with fresh ethyl acetate and dried under high vacuum to obtain 234 (797 mg, 97%) as a brown colored solid. LC-MS (Protocol B): m/z 650.3 [M+H]+, retention time=0.64 minutes. |
44% | In an iced water bath, a solution of Ac-Lys(Boc)-OH (15.2 g, 52.7 mmol) in anhydrous N ,N -dimethylformamide (250 mL) was treated with HATU (20.0 g, 52.7 mmol). After 30 minutes, L-valyl-N5-carbamoyl-N-[4-(hydroxymethyl)phenyl]-L-ornithinamide (Compound 64 in Dubowchik, G.M. et al. Bioconjug Chem 13, 855-69 (2002)5(20 g, 52.7 mmol) was added followed by N ,N -diisopropylethylamine (12.8 mL, 9.54 g, 73.8 mmol). The mixture was allowed to reach room temperature. After 2 hours, the reaction mixture was poured in MTBE to precipitate the product, which was filtered and dispersed in dichloromethane:methanol (10: 1 , 800 mL) for 20 minutes. The mixture was filtered, air dried and purified by prep-HPLC according to Method D. After concentration under reduced pressure, the residue was poured in chloroform :isopropy I alcohol 4: 1 . The solid was collected by filtration and air dried to provide the title compound 53 (15.0 g, 44%) as a white solid. HPLC (Protocol J): retention time = 3.63 minutes.1HNMR(400MHz, DMSO-d6) delta = 9.95 (s, 1 H), 8.32 (s, 1 H), 8.09 (d, J=7.5 Hz, 1 H), 8.03 (d, J=8.0 Hz, 1H), 7.72 (d, J=9O Hz. 1H), 7.57 - 7.51 (m, J=8.5 Hz, 2H), 7.27 - 7.20 (m, J=8.5 Hz. 2H), 6.77 (t, J=5.5 Hz. 1H). 5.98 (t, J=5.8 Hz, 1H), 5.43 (s, 2H), 5.11 (t, J=5.5 Hz. 1H), 443 (d. J=5.5 Hz. 2H). 4.41 - 4 35 (m. 1H). 4.29 - 4.21 (m, 1H). 4.19 (dd. J=7.o, 8.5 Hz. 1H). 3.07 - 2.92 (m. 2H). 2.91 - 2.84 (m. 2H). 2.34 (s, 1H). 1.98 (qd.J=6-7, 13.6 Hz, 1H), 1.84 (s, 3H). 1.72 - 1.50 (m, 3H), 1.43 - 1.33 (m, 12H), 1.25 (dd, J=10.3, 15.8 Hz, 3H), 0.86 (d, J=6.5 Hz, 3H), 0.83 (d, J=6.5 Hz, 3H) |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
64.52% | With pyridine; 1-ethyl-(3-(3-dimethylamino)propyl)-carbodiimide hydrochloride; at 25℃; for 1h; | To a solution of compound 1 (0.100 g, 318.09 umol, 1 eq) in pyridine (1 mL) was added EDCI (152.44mg, 795.21umol, 2.5eq) and Ac-L-Lys(Boc)-OH (119.23 mg, 413.51 umol, 1.3 eq). The mixture was stirred at 25 C for 1 hr. LC-MS showed 50% of Compound 1 remained and a peak with desired m/z (m/z: 586.1 ([M+H]+)) was detected. The residue was purified by prep- HPLC (A: H20, B: ACN). Compound 2 (0.120 g, 205.23 umol, 64.52% yield) was obtained as a white solid. Calculated MW: 584.7 observed m/z: 586.1([M+H]+) |
Tags: 23500-04-1 synthesis path| 23500-04-1 SDS| 23500-04-1 COA| 23500-04-1 purity| 23500-04-1 application| 23500-04-1 NMR| 23500-04-1 COA| 23500-04-1 structure
[ 67861-96-5 ]
Dicyclohexylamine (S)-2-((tert-butoxycarbonyl)amino)pentanoate
Similarity: 0.96
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P220 | Keep/Store away from clothing/combustible materials. |
P221 | Take any precaution to avoid mixing with combustibles |
P222 | Do not allow contact with air. |
P223 | Keep away from any possible contact with water, because of violent reaction and possible flash fire. |
P230 | Keep wetted |
P231 | Handle under inert gas. |
P232 | Protect from moisture. |
P233 | Keep container tightly closed. |
P234 | Keep only in original container. |
P235 | Keep cool |
P240 | Ground/bond container and receiving equipment. |
P241 | Use explosion-proof electrical/ventilating/lighting/equipment. |
P242 | Use only non-sparking tools. |
P243 | Take precautionary measures against static discharge. |
P244 | Keep reduction valves free from grease and oil. |
P250 | Do not subject to grinding/shock/friction. |
P251 | Pressurized container: Do not pierce or burn, even after use. |
P260 | Do not breathe dust/fume/gas/mist/vapours/spray. |
P261 | Avoid breathing dust/fume/gas/mist/vapours/spray. |
P262 | Do not get in eyes, on skin, or on clothing. |
P263 | Avoid contact during pregnancy/while nursing. |
P264 | Wash hands thoroughly after handling. |
P265 | Wash skin thouroughly after handling. |
P270 | Do not eat, drink or smoke when using this product. |
P271 | Use only outdoors or in a well-ventilated area. |
P272 | Contaminated work clothing should not be allowed out of the workplace. |
P273 | Avoid release to the environment. |
P280 | Wear protective gloves/protective clothing/eye protection/face protection. |
P281 | Use personal protective equipment as required. |
P282 | Wear cold insulating gloves/face shield/eye protection. |
P283 | Wear fire/flame resistant/retardant clothing. |
P284 | Wear respiratory protection. |
P285 | In case of inadequate ventilation wear respiratory protection. |
P231 + P232 | Handle under inert gas. Protect from moisture. |
P235 + P410 | Keep cool. Protect from sunlight. |
Response | |
Code | Phrase |
P301 | IF SWALLOWED: |
P304 | IF INHALED: |
P305 | IF IN EYES: |
P306 | IF ON CLOTHING: |
P307 | IF exposed: |
P308 | IF exposed or concerned: |
P309 | IF exposed or if you feel unwell: |
P310 | Immediately call a POISON CENTER or doctor/physician. |
P311 | Call a POISON CENTER or doctor/physician. |
P312 | Call a POISON CENTER or doctor/physician if you feel unwell. |
P313 | Get medical advice/attention. |
P314 | Get medical advice/attention if you feel unwell. |
P315 | Get immediate medical advice/attention. |
P320 | |
P302 + P352 | IF ON SKIN: wash with plenty of soap and water. |
P321 | |
P322 | |
P330 | Rinse mouth. |
P331 | Do NOT induce vomiting. |
P332 | IF SKIN irritation occurs: |
P333 | If skin irritation or rash occurs: |
P334 | Immerse in cool water/wrap n wet bandages. |
P335 | Brush off loose particles from skin. |
P336 | Thaw frosted parts with lukewarm water. Do not rub affected area. |
P337 | If eye irritation persists: |
P338 | Remove contact lenses, if present and easy to do. Continue rinsing. |
P340 | Remove victim to fresh air and keep at rest in a position comfortable for breathing. |
P341 | If breathing is difficult, remove victim to fresh air and keep at rest in a position comfortable for breathing. |
P342 | If experiencing respiratory symptoms: |
P350 | Gently wash with plenty of soap and water. |
P351 | Rinse cautiously with water for several minutes. |
P352 | Wash with plenty of soap and water. |
P353 | Rinse skin with water/shower. |
P360 | Rinse immediately contaminated clothing and skin with plenty of water before removing clothes. |
P361 | Remove/Take off immediately all contaminated clothing. |
P362 | Take off contaminated clothing and wash before reuse. |
P363 | Wash contaminated clothing before reuse. |
P370 | In case of fire: |
P371 | In case of major fire and large quantities: |
P372 | Explosion risk in case of fire. |
P373 | DO NOT fight fire when fire reaches explosives. |
P374 | Fight fire with normal precautions from a reasonable distance. |
P376 | Stop leak if safe to do so. Oxidising gases (section 2.4) 1 |
P377 | Leaking gas fire: Do not extinguish, unless leak can be stopped safely. |
P378 | |
P380 | Evacuate area. |
P381 | Eliminate all ignition sources if safe to do so. |
P390 | Absorb spillage to prevent material damage. |
P391 | Collect spillage. Hazardous to the aquatic environment |
P301 + P310 | IF SWALLOWED: Immediately call a POISON CENTER or doctor/physician. |
P301 + P312 | IF SWALLOWED: call a POISON CENTER or doctor/physician IF you feel unwell. |
P301 + P330 + P331 | IF SWALLOWED: Rinse mouth. Do NOT induce vomiting. |
P302 + P334 | IF ON SKIN: Immerse in cool water/wrap in wet bandages. |
P302 + P350 | IF ON SKIN: Gently wash with plenty of soap and water. |
P303 + P361 + P353 | IF ON SKIN (or hair): Remove/Take off Immediately all contaminated clothing. Rinse SKIN with water/shower. |
P304 + P312 | IF INHALED: Call a POISON CENTER or doctor/physician if you feel unwell. |
P304 + P340 | IF INHALED: Remove victim to fresh air and Keep at rest in a position comfortable for breathing. |
P304 + P341 | IF INHALED: If breathing is difficult, remove victim to fresh air and keep at rest in a position comfortable for breathing. |
P305 + P351 + P338 | IF IN EYES: Rinse cautiously with water for several minutes. Remove contact lenses, if present and easy to do. Continue rinsing. |
P306 + P360 | IF ON CLOTHING: Rinse Immediately contaminated CLOTHING and SKIN with plenty of water before removing clothes. |
P307 + P311 | IF exposed: call a POISON CENTER or doctor/physician. |
P308 + P313 | IF exposed or concerned: Get medical advice/attention. |
P309 + P311 | IF exposed or if you feel unwell: call a POISON CENTER or doctor/physician. |
P332 + P313 | IF SKIN irritation occurs: Get medical advice/attention. |
P333 + P313 | IF SKIN irritation or rash occurs: Get medical advice/attention. |
P335 + P334 | Brush off loose particles from skin. Immerse in cool water/wrap in wet bandages. |
P337 + P313 | IF eye irritation persists: Get medical advice/attention. |
P342 + P311 | IF experiencing respiratory symptoms: call a POISON CENTER or doctor/physician. |
P370 + P376 | In case of fire: Stop leak if safe to Do so. |
P370 + P378 | In case of fire: |
P370 + P380 | In case of fire: Evacuate area. |
P370 + P380 + P375 | In case of fire: Evacuate area. Fight fire remotely due to the risk of explosion. |
P371 + P380 + P375 | In case of major fire and large quantities: Evacuate area. Fight fire remotely due to the risk of explosion. |
Storage | |
Code | Phrase |
P401 | |
P402 | Store in a dry place. |
P403 | Store in a well-ventilated place. |
P404 | Store in a closed container. |
P405 | Store locked up. |
P406 | Store in corrosive resistant/ container with a resistant inner liner. |
P407 | Maintain air gap between stacks/pallets. |
P410 | Protect from sunlight. |
P411 | |
P412 | Do not expose to temperatures exceeding 50 oC/ 122 oF. |
P413 | |
P420 | Store away from other materials. |
P422 | |
P402 + P404 | Store in a dry place. Store in a closed container. |
P403 + P233 | Store in a well-ventilated place. Keep container tightly closed. |
P403 + P235 | Store in a well-ventilated place. Keep cool. |
P410 + P403 | Protect from sunlight. Store in a well-ventilated place. |
P410 + P412 | Protect from sunlight. Do not expose to temperatures exceeding 50 oC/122oF. |
P411 + P235 | Keep cool. |
Disposal | |
Code | Phrase |
P501 | Dispose of contents/container to ... |
P502 | Refer to manufacturer/supplier for information on recovery/recycling |
Physical hazards | |
Code | Phrase |
H200 | Unstable explosive |
H201 | Explosive; mass explosion hazard |
H202 | Explosive; severe projection hazard |
H203 | Explosive; fire, blast or projection hazard |
H204 | Fire or projection hazard |
H205 | May mass explode in fire |
H220 | Extremely flammable gas |
H221 | Flammable gas |
H222 | Extremely flammable aerosol |
H223 | Flammable aerosol |
H224 | Extremely flammable liquid and vapour |
H225 | Highly flammable liquid and vapour |
H226 | Flammable liquid and vapour |
H227 | Combustible liquid |
H228 | Flammable solid |
H229 | Pressurized container: may burst if heated |
H230 | May react explosively even in the absence of air |
H231 | May react explosively even in the absence of air at elevated pressure and/or temperature |
H240 | Heating may cause an explosion |
H241 | Heating may cause a fire or explosion |
H242 | Heating may cause a fire |
H250 | Catches fire spontaneously if exposed to air |
H251 | Self-heating; may catch fire |
H252 | Self-heating in large quantities; may catch fire |
H260 | In contact with water releases flammable gases which may ignite spontaneously |
H261 | In contact with water releases flammable gas |
H270 | May cause or intensify fire; oxidizer |
H271 | May cause fire or explosion; strong oxidizer |
H272 | May intensify fire; oxidizer |
H280 | Contains gas under pressure; may explode if heated |
H281 | Contains refrigerated gas; may cause cryogenic burns or injury |
H290 | May be corrosive to metals |
Health hazards | |
Code | Phrase |
H300 | Fatal if swallowed |
H301 | Toxic if swallowed |
H302 | Harmful if swallowed |
H303 | May be harmful if swallowed |
H304 | May be fatal if swallowed and enters airways |
H305 | May be harmful if swallowed and enters airways |
H310 | Fatal in contact with skin |
H311 | Toxic in contact with skin |
H312 | Harmful in contact with skin |
H313 | May be harmful in contact with skin |
H314 | Causes severe skin burns and eye damage |
H315 | Causes skin irritation |
H316 | Causes mild skin irritation |
H317 | May cause an allergic skin reaction |
H318 | Causes serious eye damage |
H319 | Causes serious eye irritation |
H320 | Causes eye irritation |
H330 | Fatal if inhaled |
H331 | Toxic if inhaled |
H332 | Harmful if inhaled |
H333 | May be harmful if inhaled |
H334 | May cause allergy or asthma symptoms or breathing difficulties if inhaled |
H335 | May cause respiratory irritation |
H336 | May cause drowsiness or dizziness |
H340 | May cause genetic defects |
H341 | Suspected of causing genetic defects |
H350 | May cause cancer |
H351 | Suspected of causing cancer |
H360 | May damage fertility or the unborn child |
H361 | Suspected of damaging fertility or the unborn child |
H361d | Suspected of damaging the unborn child |
H362 | May cause harm to breast-fed children |
H370 | Causes damage to organs |
H371 | May cause damage to organs |
H372 | Causes damage to organs through prolonged or repeated exposure |
H373 | May cause damage to organs through prolonged or repeated exposure |
Environmental hazards | |
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 |
Sorry,this product has been discontinued.
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