ࡱ > _ a ^ E@ ` bjbj 8f % T T T T T T T h 0 0 0 8 h | h R " . . . a a a $ p R " p T ] a T T . . / b b b T . T . b b ( b T T . Gg 0 6 4 " 0 R , 2# X 2# h h T T T T 2# T a ' D b k o a a a h h $ @ " h h # Supplementary Material (ESI) for Chemical Communications # This journal is The Royal Society of Chemistry 2004 Electronic Supplementary Information Synthesis and vesicle formation of a nido-carborane cluster lipid for boron neutron capture therapy Hiroyuki Nakamura,*a Yusuke Miyajima,a Toshiaki Takeib Satoshi Kasaokac and Kazuo Maruyamac a Department of Chemistry, Faculty of Science, Gakushuin University, Tokyo 171-8588, Japan. Fax: 81 3 5992 1029; Tel: 81 3 3986 0221, ext. 6491; E-mail: hiroyuki.nakamura@gakushuin.ac.jp b Institute for Molucular Biology, Gakushuin University, Tokyo 171-8588, Japan c Department of Biopharmaceutics, School of Pharmaceutical Sciences, Teikyo University, Kanagawa, 199-0195, Japan Synthetic Experiments General: 1H NMR and 13C NMR spectra were measured on a JEOL JNM-AL 300 (300 MHz) and VARIAN UNITY-INOVA 400 (400 MHz) spectrometers. Chemical shifts of 1H NMR were expressed in parts per million downfield from CDCl3 as an internal standard ( d = 7 . 2 4 ) i n C D C l 3 . C h e m i c a l s h i f t s o f 1 3 C N M R w e r e e x p r e s s e d i n p a r t s p e r m i l l i o n d o w n f i e l d f r o m C D C l 3 a s a n i n t e r n a l s t a n d a r d ( d = 7 7 . 0 ) i n C D C l 3 . I R s p e c t r a w e r e m e a s u r e d o n a S h i m a d z u F T I R - 8 2 0 0 A S p e c t r o m e t e r . A n a l y t i c a l t h i n l a y e r c h r o m a t o g r a p h y ( T L C ) was performed on a glass plates (Merck Kieselgel 60 F254, layer thickness 0.2 mm). Visualization was accompanied by KMnO4. Column chromatography was performed on silica gel (Merck Kieselgel 70-230 mesh). All reactions were carried out under argon atmosphere using standard Schlenk techniques. Most chemicals and solvents were analytical grade and used without further purification. 3-Heptadecyloxy-2-heptadecyloxymethyl-1-propene (3); To a NaH (16.0 g, 0.340 mol) suspension in THF (100 mL) was added hexadecanol (26.9 g, 0.105 mol) in THF (100 mL) dropwise at 0C under Ar. The temperature was allowed to room temperature and the reaction mixture was stirred for 4 h. The mixture was again cooled to 0C and 2-chloromethyl-3-chloro-1-propene (5.5 mL, 47.6 mmol) was added dropwise. The reaction mixture was stirred at room temperature for 1 h and then refluxed for 89 h. The reaction was quenched with saturated aqueous NH4Cl solution (50 mL) and the mixture was extracted with ether, washed with saturated aqueous NaCl s o l u t i o n , d r i e d o v e r a n h y d r o u s M g S O 4 , a n d t h e n c o n c e n t r a t e d . P u r i f i c a t i o n b y s i l i c a g e l c o l u m n c h r o m a t o g r a p h y ( H e x a n e : e t h y l a c e t a t e = 5 0 : 1 ) g a v e 3 ( 2 5 . 1 g , 4 4 . 3 m m o l , 9 3 % y i e l d ) a s a w h i t e s o l i d : 1 H N M R ( C D C l 3 , 4 0 0 M H z ) 5 . 1 4 ( s , 2 H ) , 3 . 9 4 ( s , 4 H ) , 3 . 3 8 ( t , J = 6 . 8 H z , 4 H ) , 1 . 5 5 ( m , 4 H ) , 1 . 2 3 ( b r s , 5 6 H ) , 0 . 8 6 ( t , J = 7 . 2 H z , 6 H ) ; 1 3 C N M R ( C D C l 3 , 1 0 0 M H z ) 1 4 3 . 3 , 1 1 3 . 3 , 7 1 . 5 , 7 0 . 6 , 3 1 . 9 , 2 9 . 7 , 2 9 . 7 , 2 9 . 6 , 2 9 . 5 , 2 9 . 4 , 2 6 . 2 , 2 2 . 7 , 1 4 . 1 ; I R ( K B r ) 2 9 1 6 , 2 8 4 8 , 2 8 0 4 , 2 7 3 1 , 1 6 6 6 , 1 4 7 3 , 1 3 8 0 , 1 1 0 9 , 1 0 7 8 , 1 0 4 5 , 1 0 1 0 c m - 1 ; H R M S ( E S I ) m / z ( M + N a ) + c a l c d f o r C 3 8 H 7 6 O 2 N a 5 8 7 . 5 7 4 3 , f o u n d 5 8 7 . 5 7 9 5 . 3 - H e p t a d e c y l o x y - 2 - h e p t a d e c y l o x y m e t h y l - p r o p a n e - 1 - o l ( 4 ) ; T o a s o l u t i o n o f 3 ( 1 8 . 1 g , 3 1 . 9 m m o l ) i n T H F ( 3 0 0 m L ) w a s a d d e d B H 3 eM e 2 S ( 3 1 . 9 m m o l ) d r o p w i s e a t 0 C u n d e r A r . T h e t e m p e r a t ure was allowed to room temperature and the reaction mixture was stirred for 6 h. The mixture was again cooled to 0C and an aqueous NaOH solution (3 M, 100 mL) was added. The reaction mixture was stirred at0C for 24 h and then H2O2 (30%, 100 mL) was added. After 12 h at room temperature with stirring, K2CO3 (5g) was added and the mixture was extracted with ether, washed with saturated aqueous NaCl solution, dried over anhydrous MgSO4, and then concentrated. The residue was passed through short column chro m a t o g r a p h y o n S i O 2 w i t h h e x a n e / e t h y l a c e t a t e ( 3 0 / 1 ) a n d t h e n r e c r y s t a l i z a t i o n f r o m h e x a n e g a v e 4 ( 1 3 . 3 g , 2 2 . 8 m m o l , 7 1 % y i e l d ) a s a c o l o r l e s s c r y s t a l : 1 H N M R ( C D C l 3 , 4 0 0 M H z ) 3 . 7 4 ( t , J = 5 . 2 H z , 2 H ) , 3 . 5 0 ( m , 4 H ) , 3 . 3 8 ( t , J = 5 . 2 H z , 4 H ) , 2 . 0 7 ( q u i n t , J = 5 . 6 H z , 1 H ) , 1 . 5 3 ( t , J = 6 . 4 H z , 4 H ) , 1 . 2 3 ( b r s , 5 6 H ) , 0 . 8 6 ( t , J = 6 . 8 H z , 6 H ) ; 1 3 C N M R ( C D C l 3 , 1 0 0 M H z ) 7 1 . 8 , 7 1 . 2 , 6 5 . 1 , 4 1 . 4 , 3 2 . 1 , 2 9 . 9 , 2 9 . 8 , 2 9 . 7 , 2 9 . 6 , 2 6 . 3 , 2 2 . 9 , 1 4 . 3 ; I R ( K B r ) 3 3 2 8 , 2 8 3 9 , 2 7 4 4 , 1 4 7 1 , 1 3 7 7 , 1 3 1 3 , 1 2 3 2 , 1 2 1 3 , 1 1 9 1 , 1 1 1 6 , 1 0 4 1 cm-1; HRMS (ESI) m/z (M+Na)+ calcd for C38H78O3Na 605.5848, found 605.5816. 3-Heptadecyloxy-2-heptadecyloxymethyl-1-propargyloxypropane (5); To a NaH (1.03 g, 25.7 mmol) suspension in THF (20 mL) was added 4 (3.0 g, 5.2 mmol) in THF (15 mL) dropwise at 0C under Ar. The temperature was allowed to room temperature and the reaction mixture was stirred for 3 h. The mixture was again cooled to 0C and propargyl bromide (1.6 mL, 15.4 mmol) was added dropwise. The reaction mixture was stirred at room temperature for 20 h. The reaction was quenched with saturated aqueous NH4Cl solution (20 mL) and the mixture was extracted with ether, washed with saturated aqueous NaCl solution, dried over anhydrous MgSO4, and then concentrated. Purification by silica gel column chromatography (Hexane:ethyl acetate = 30:1) gave 5 (1.87 g, 3.0 mmol, 58% yield) as a yellow solid: 1HNMR (CDCl3, 400MHz) 4 . 4 1 ( d , J = 2 . 4 H z , 2 H ) , 3 . 5 5 ( d , J = 6 . 0 H z , 2 H ) , 3 . 4 3 ( d , J = 6 . 4 H z , 4 H ) , 3 . 3 8 ( t , J = 6 . 4 H z , 4 H ) , 2 . 3 9 ( s , 1 H ) , 2 . 1 4 ( m , 1 H ) , 1 . 5 2 ( t , J = 6 . 8 H z , 4 H ) , 1 . 2 4 ( b r s , 5 6 H ) , 0 . 8 7 ( t , J = 6 . 8 H z , 6 H ) ; 1 3 C N M R ( C D C l 3 , 1 0 0 M H z ) 7 4 . 2 , 7 1 . 4 , 6 9 . 2 , 6 9 . 0 , 5 8 . 6, 40.4, 32.1, 29.9, 29.8, 29.7, 29.6, 26.4, 22.9, 14.3; IR (KBr) 3249, 2912, 2848, 2798, 1508, 1473, 1458, 1359, 1087 cm-1; HRMS (ESI) m/z (M+Na)+ calcd for C41H80O3Na 643.6005, found 643.5989. (3-Heptadecyloxy-2-heptadecyloxymethyl-1-propoxy)methyl-ortho-carborane (6); The mixture of 5 (0.61 g, 0.99 mmol), decaborane (0.15 g, 1.2 mmol), and acetonitrile (1.56 mL, 29.6 mmol) in toluene (20 mL) was stirred under reflux condition for 6 h. The solvent was removed under the reduced pressure and the residue wa s p u r i f i e d b y s i l i c a g e l c o l u m n c h r o m a t o g r a p h y ( H e x a n e : e t h y l a c e t a t e = 3 0 : 1 ) t o g i v e 6 ( 0 . 6 1 g , 0 . 8 3 m m o l , 8 0 % y i e l d ) a s a w h i t e s o l i d : 1 H N M R ( C D C l 3 , 4 0 0 M H z ) 3 . 9 7 ( b r s , 1 H ) , 3 . 5 1 ( d , J = 5 . 6 H z , 2 H ) , 3 . 4 0 - 1 . 4 0 ( m , 1 0 H ) , 2 . 0 9 ( q u i n t , J = 6 . 0 H z , 1 H ) , 1 . 5 1 ( t , J = 6 . 8 H z , 4 H ) , 1 . 2 4 ( b r s , 5 6 H ) , 0 . 8 6 ( t , J = 6 . 8 H z , 6 H ) ; 1 3 C N M R ( C D C l 3 , 1 0 0 M H z ) 7 2 . 3 , 7 1 . 6 , 7 0 . 4 , 6 8 . 9 , 5 7 . 7 , 4 0 . 4 , 3 2 . 1 , 2 9 . 9 , 2 9 . 8 , 2 9 . 7 , 2 9 . 6 , 2 6 . 4 , 2 2 . 9 , 1 4 . 3 ; I R ( K B r ) 2 9 1 8 , 2 8 5 0 , 2 5 7 2 , 1 4 6 5 , 1 3 7 7 , 1 1 1 2 , 1 0 9 5 , 1 0 1 2 c m - 1 ; M S ( M A L D I ) m / z ( M + N a)+ C41H90O3B10Na 763.8. (3-Heptadecyloxy-2-heptadecyloxymethyl-1-propoxy)methyl-ortho-nido-carborane sodium salt (2); The mixture of 6 (0.92 g, 1.25 mmol), and NaOMe (0.68 g, 12.5 mmol) in MeOH (15 mL) was stirred under reflux condition for 14 h. The reaction mixture was diluted with water (20 mL), extracted with ethyl acetate, washed with saturated aqueous NaCl solution, dried over anhydrous MgSO4, and then concentrated. Purification by silica gel column chromatography (gradient from hexane:ethyl acetate = 3 0 : 1 t o e t h y l a c e t a t e o n l y ) g a v e 2 ( 0 . 5 6 g , 0 . 7 2 m m o l , 5 7 % y i e l d ) a s a w h i t e s o l i d : 1 H N M R ( C D C l 3 , 4 0 0 M H z ) 3 . 7 7 ( d , J = 5 . 6 H z , 2 H ) , 3 . 5 2 ( m , 6 H ) , 3 . 4 0 ( t , J = 6 . 4 H z , 4 H ) , 2 . 0 9 ( m , 1 H ) , 1 . 5 4 ( m , 4 H ) , 1 . 2 4 ( b r s , 5 6 H ) , 0 . 8 7 ( t , J = 6 . 8 H z , 6 H ) ; 1 3 C N M R ( C r t $ % & ʿxfQxCQ1 #h4rJ h4rJ 5CJ aJ mH nH tHh4rJ h4rJ 5CJ PJ aJ (h4rJ h4rJ 56CJ H*aJ mH nH u "h4rJ h4rJ 5CJ aJ mH nH u h4rJ h4rJ 5CJ aJ h4rJ h4rJ OJ QJ aJ h4rJ h;k 5CJ OJ QJ aJ h4rJ h4rJ 5CJ OJ QJ aJ h4rJ h4rJ 5OJ QJ o( h;k 5OJ QJ o( h4rJ h;k 5OJ QJ h4rJ 5OJ QJ h4rJ 5OJ QJ o( hZF 5OJ QJ hZF hZF 5OJ QJ ; s t [ f N 0 d UD ]0 gd4rJ d gd4rJ d gd4rJ d ] gd4rJ ] ^ gd4rJ ] ^ gd4rJ d gd4rJ $d a$gd4rJ $d a$gdZF ` ` & 4 5 E F G Y Z [ \ ] 8 N ^ f õà~w~gWWWWWOw~g h4rJ h4rJ o(h4rJ h4rJ mH nHo(tHu h4rJ h4rJ ]mH nH o(tH h4rJ h4rJ h4rJ h4rJ ]h4rJ h4rJ mH nH u h4rJ h4rJ H*mH nH u )h4rJ h4rJ 56CJ aJ mH nHtHu h4rJ h4rJ 5CJ H*aJ ,h4rJ h4rJ 56CJ H*]aJ mH nH tH ,h4rJ h4rJ 56CJ H*aJ mH nHtHu h4rJ h4rJ 5CJ PJ aJ f g " $ N R $ & 5 8 x y μvvj^ h4rJ hT