(1 ), the prep job of palmitoyl chloride (Pal-Cl).
In a 250mL single-neck round-bottom flask, include 20.52 g palmitic acid, include 60mL thionyl chloride to liquify, reflux at 80 ° C for 5 hrs, cool to location temperature level, as well as also revolve dry the solvent to obtain the white oily target item palmitoyl chloride (21.6 g, 98%). 1HNMR( CDCl3,300 MHz) δ0.86 -0.90( t,3 H),1.26 -1.34( m,24 H),1.66 -1.75( m,2 H),2.86 -2.90( t,2 H).
( 2) Prep Work of Pal-Lys( Cbz)- OH.
500mL single-neck round base flask, weigh 21g Na2CO3 as well as dissolve it in 120mL water, consist of 60mL of tetrahydrofuran, after that add 11g of Nε-Cbz-L-lysine to dissolve, at room temperature, include 60mL of tetrahydrofuran treatment consisting of 11.2 g of palmitoyl chloride dropwise, and likewise the addition is complete After that, the feedback was blended for 16h. After the reaction, the pH of the system was gotten used to 3-4 with 1M HCl, removed with dichloromethane, dried over anhydrous sodium sulfate, filteringed system, as well as the filtrate was spin-dried to get the white strong target product Pal-Lys( Cbz)- OH (19.3 g, 93%). 1HNMR( DMSO-d6,300 MHz) δ0.83 -0.87( m,3 H),1.23 -1.69( m,34 H),2.94 -3.00( m,2 H),4.10 -4.17( m,1 H),4.57( s, 1H), 5.00( s, 2H), 7.23-7.39( m, 5H), 7.99-8.01( d, 1H), 12.43( s, 1H).
( 3) Preparation Work of Pal-Lys( Cbz)- ONHS triggered ester.
In a 500mL single-neck round-bottomed flask, liquify 19.3 g Pal-Lys( Cbz)- OH in 300mL dichloromethane, add 6.4 g N-hydroxysuccinimide (NHS), 0.91 g DMAP as well as 10.7 g EDCl under ice restroom troubles, along with coating adding Later on, it was slowly gone back to space temperature level as well as blended for 18 h. After the action, the pH of the system was adjusted to 3-4 with 1M HCl, extracted with dichloromethane, dried over anhydrous sodium sulfate, filteringed system, as well as the filtrate was spin-dried to acquire the white strong target product Pal-Lys( Cbz)- ONHS (19.4 g, 85%). 1HNMR( DMSO-d6,300 MHz) δ0.82 -0.87( m,3 H),1.23 -1.29( m,26 H),1.40 -1.50( m,4 H),1.71 -1.86( m,2 H),2.10 -2.15( m, 2H), 2.79( s, 4H), 2.95-2.99( m, 2H), 4.54-4.61( m, 1H), 5.00( s, 2H), 7.24-7.39( m, 5H), 8.42-8.44( d, 1H).
( 4) Preparation Job of Pal-Lys( Cbz)- Val-OH.
500mL single-neck round bottom flask, consider 11.7 g Na2CO3 as well as liquify it in 120mL water, include 3.9 g L-valine to liquify, under nitrogen feel, under ice restroom conditions, decline 120mL of tetrahydrofuran treatment including 17g Pal-Lys( Cbz)- ONHS, dropwise add After final thought, it was gradually gone back to location temperature level, and likewise the response was mixed for 20h. After the reaction, the pH of the system was adapted to 3-4 with 1M HCl, eliminated with ethyl acetate, dried out over anhydrous salt sulfate, filteringed system, in addition to the filtrate was spin-dried to get the white solid target thing Pal-Lys( Cbz)- Val-OH (16.5 g, 97%). 1HNMR( DMSO-d6,300 MHz) δ0.82 -0.87( m,9 H),1.22 -1.46( m,32 H),1.98 -2.12( m,3 H),2.93 -2.97( m,2 H),4.09 -4.17( m,1 H),4.30 -4.40( m,1 H),4.99( s,2 H),7.23 -7.39( m,5 H),7.84 -7.91( m,1 H),7.97 -8.01( m,1 H),12.55( s,1 H).
( 5) Preparation of Pal-Lys( Cbz)- Val-ONHS activated ester.
In a 500mL single-neck round-bottom flask, liquify 17.5 g Pal-Lys( Cbz)- Val-OH in 200mL dichloromethane, include 4.9 g N-hydroxysuccinimide (NHS), 0.69 g DMAP and additionally 8.17 g EDCl under ice restroom problems, After the enhancement, it was progressively gone back to space temperature level and stirred for 18h. After the response, the pH of the system was adjusted to 3-4 with 1M HCl, drawn out with dichloromethane, dried out over anhydrous sodium sulfate, filteringed system, and additionally the filtrate was spin-dried to acquire the white solid target item Pal-Lys( Cbz)- Val-ONHS (18.2 g, 90%). 1HNMR( DMSO-d6,300 MHz) δ0.82 -1.02( m,9 H),1.22 -1.57( m,32 H),1.98 -2.21( m,3 H),2.80( s,4 H),2.91 -2.97( m, 2H), 4.31-4.63( m, 2H), 4.99( s, 2H), 7.22-7.39( m, 5H), 7.88-7.95( m, 1H), 8.41-8.51( m, 1H).
( 6) Prep Work of Pal-Lys( Cbz)- Val-Lys( Cbz)- OH.
500mL single-neck round-bottom flask, review 11.7 g Na2CO3 and likewise liquify it in 120mL water, include 60mL tetrahydrofuran, after that include 9.3 g Nε-Cbz-L-Lysine to dissolve, at area temperature, drop right into 60mL having 19.7 g Pal-Lys( Cbz)- Val -ONHS solution in tetrahydrofuran, after the dropwise addition, the action was blended for 16h. After the response, the pH of the system was adjusted to 3-4 with 1M HCl, removed with dichloromethane, dried out over anhydrous salt sulfate, filteringed system, and also the filtrate was spin-dried to acquire the white strong target thing Pal-Lys( Cbz)- Val-Lys( Cbz )- OH (23.56 g, 97%). 1HNMR( DMSO-d6,300 MHz) δ0.78 -0.85( m,9 H),1.22 -1.59( m,38 H),1.96 -2.09( m,3 H),2.91 -2.99( m,4 H),4.12 -4.32( m,3 H),4.99( s,4 H),7.21 -7.38( m,10 H),7.60 -8.16( m,3 H),12.49( s,1 H).
( 7) Prep Work of Pal-Lys-Val-Lys-OH (palmitoyl tripeptide-8).
In a 2000mL single-neck round base flask, 23.56 g Pal-Lys( Cbz)- Val-Lys( Cbz)- OH was liquified in 1500mL methanol, and 4.7 g 5% wet Pd/C( Pal-Lys( Cbz)- Val-Lys( Cbz) was included The mass proportion of )- OH to Pd/C was 5:1), nitrogen was changed three times, and also hydrogen was transformed 3 times. Finally, the hydrogen ambience was preserved, along with the reaction was mixed at space temperature as well as normal stress for 24-hour. After the response, Pd/C was filtered off, the methanol was spin-dried, and afterwards recrystallized from ethanol to obtain the target product palmitoyl tripeptide-8 (13.1 g, 80%) as a white powder. 1HNMR( CD3OD,300 MHz) δ0.87 -0.98( m,9 H),1.29 -1.86( m,40 H),2.12 -2.31( m,3 H),2.80 -2.88( m,4 H),4.12 -4.46( m, 3H).
