마이크로파를이용한간담도계영상방사성의약품 99m Tc-Mebrofenin 의합성

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1 Journal of the Korean Chemical Society 2005, Vol. 49, No. 4 Printed in the Republic of Korea 마이크로파를이용한간담도계영상방사성의약품 Tc-Mebrofenin 의합성 «Á yá x*á w ( ) Microwave assisted Synthesis of Tc-Mebrofenin for Imaging Hepatobiliary System Hui Jeong Gwon, Seung Ho Jang, Sang Hyun Park*, and Byung Chul Shin Division of Radioisotope Production and Application, Korea Atomic Energy Research Institute, Daejeon , Korea (Received April 20, 2005). w w yw» w yw r ( Tc-mebrofenin) j q w wš, w, w z w w wš w. x w š r w w, j q w lj p r t w t 99.5% t z. : r, y, j q ABSTRACT. Tc-Labeled N-(3-bromo-2,4,6-trimethylacetanilido)-iminodiacetic acid ( Tc-mebrofenin) has been used to assess the functional integrity of the hepatobiliary system. A fast and convenient procedure was established for the synthesis of mebrofenin and radiochemical labeling with Tc. 3-Bromo-2,4,6-trimethylacetanilide was synthesized in large scale under microwave irradiation (300 W, 5 min) by acylation (60% yield) and bromination (96% yield). And it was treated with the disodium salt of iminodiacetic acid under microwave irradiation (300 W, 20 min) to afford mebrofenin in 86% yield. The attractive feature in this procedure involves microwave irradiation that results in shorter reaction times, higher yields, cleaner reaction products and an easier work-up than classical heating. The synthesis time of 2,4,6- trimethylacetanilide was reduced from 1 h to 5 min; the synthesis time of 3-bromo-2,4,6-trimethylacetanilide was reduced from 24 h to 5 min; the synthesis time of mebrofenin was reduced from 48 h to 20 min, when compared with that of the conventional method. Iminodiacetic acid of mebrofenin is a metal complexing moiety that strongly binds Tc. Tc-Mebrofenin between the reduced technetium, prepared by a stannous reduction of the pertechnetate ion, and mebrofenin was formed under microwave irradiation (300 W, 1 min). The synthesis time of Tc-mebrofenin was reduced from 30 min to 1 min, when compared with that of the conventional method. 99.5% of Labeling efficiency (LE) of Tc-mebrofenin was determined by using radio-tlc. Keywords: Mebrofenin, Hepatobiliary Imaging, Microwave Irradiation 370

2 2005, Vol. 49, No. 4 j q w 131 I t (Rose Bengal) l Tc t p (iminodiacetic acid, IDA)»¾ yw y. Tc 1,2 w w p,»(6 h)» w (140 kev) w w t ƒ š w. w Tc-IDA ƒ ü» s š, 3-7, N-(3-bromo-2,4,6- trimethylacetanilido)iminodiacetic acid(mebrofenin) ƒ ù p š. 8 w y w j p yw A w ƒ p B ey (Fig. 1). ù,» w w.,» ey 48 ƒ š, 20% w. 3 w» w š z w v ƒ., p(dipole moment) ƒ w j q(microwave) ƒ ƒ» š.» j q š, y ey yw w. 9-18, w w yw» w yw r (mebrofenin) Fig. 1. A) Chloroacetyl compound, B) Sodium salt of iminodiacetic acid. Tc-Mebrofenin w 371 wš, w, w z w w wš w, j q w r x w š w w. š j q w r w. p 2,4,6-trimethylaniline chloroacetyl chloride w 58% yw 1 w w, j q w 60% w w. w, 1 ù, j q w 5 z». w yw 1 w k w 16 w 92% yw 2 w w. j q w 5 96% w w. yw 2 iminodiacetic acid, disodium salt hydrate w k w w g 52% yw 3 w w., j q w w { 20 86% w w. ƒƒ Table 1 w ùkü. Tc- r. p B w w» w w y. lj p j q w w r 4 w lj p SnCl2 2H2O w y w. 4 w Tc- r t p w» w w d j m v Table 2 ùkü. yùp sy w ITLC- SA lj p R f , 4 R f % ùkü. w, acetonitrile:water(3:1, v/v) yw w ITLC-SG lj p- g

3 372 «Á yá xá Table 1. Comparison of microwave irradiation method with conventional heating method of sequences for the N-(3-bromo-2,4,6- trimethylacetanilido) iminodiacetic acid (mebrofenin) Product Reaction time Yield (%) Microwave Conventional Microwave Conventional 1 5 min 2 h min 16 h min 18 h Table 2. Radio-TLC analysis of Tc-N-(3-bromo-2,4,6-trimethylacetanilido) iminodiacetic acid ( Tc-mebrofenin). Chromatographic Rf value of Tc species System Unreduced Tc Reduced, hydrolyzed Tc Tc-mebrofenin Support (ITLC-SA/SG) / / / Area value (at ph=5) 0% 0.05% 99.5% (=LE) a a Labeling efficiency (LE) was determined by the equation of [100%-(% of unreduced Tc + % of reduced, hydrolyzed Tc)] Fig. 2. Labeling efficiency of Tc-mebrofenin synthesized by using microwave irradiation: influence of ph and reaction time. Keys: (ph 3), (ph 5), (ph 7), (ph 9). Rf % ù kü š, 4 R f % ùkü. w t t t z t ƒƒ yw j m w. x w w lj p, lj p- g,» w ƒƒ yw Rf t z w. ƒƒ ph t, ph % ƒ t z ùkü š, ph ƒ ú t z y w (Fig. 2). p ƒ y lj p w y x w,» y lj p y lj p- g g x w t z ƒ. l, j q w w r lj p t w Tc- r yw p ù ù w ƒ š y w. t sodium pertechnetate (Na TcO 4) 0.9% w 99 Mo/ Tc-generator (Unitech 500, ( ) l, ) l w w. j q w w e š z» ƒ MicroSYNTH(Milestone, Sorisole, Italy) w. d w Mel-Temp(50/60 cycles, volts, 250 watts) w, yw w Bomen MB154 FTIR (KBr pellets or neat), Bruker 300-MHz NMR Spectrometer(w» w, ), Varian MAT 371 Mass Spectrometer (70 ev) w. t yw t z d w» w gamma-ray Scanner (RITA TLC Analysis, x Journal of the Korean Chemical Society

4 j q w Tc-Mebrofenin w 373 Scheme 1. Conventional synthesis of N-(3-bromo-2,4,6-trimethylacetanilido) iminodiacetic acid (mebrofenin). Scheme 2. Synthesis of N-(3-bromo-2,4,6-trimethylacetanilido) iminodiacetic acid (mebrofenin) using conventional heating. Raytest, Germany)ƒ ITLC(Instant Thin-Layer Chromatography) Scanner (EC & G Berthold linear Analyzer, Germany) w. Silica gel s ITLC-SG(Merck, Darmstadt, Germany) polysilicic acidƒ s ITLC-SA(Merck, Darmstadt, Germany) w. x p 1 w.» w w ( A). w w Scheme 1 w ùkü. ƒ w w ( B). w w Scheme 2 w ùkü. 2,4,6-Trimethylacetanilide(1) w. w 150 ml ƒ v j 4.21 ml(30 mmol) 2,4,6-trimethylaniline š 30 ml p (acetic acid) z, ml(33 mmol) chloroacetyl chloride. 1 w z, 42 ml 5.9 g sodium acetate, trihydrate v j e g. e wš k z, k w 2,4,6-trimethylacetanilide (1) w w. 2,4,6-Trimethylacetanilide (1): mp 184 o C; IR(KBr) 3236, 3038, 1671, 1608, 1540, 1485, 1333, 1241, 1202, 1039 cm 1 ; 1 H NMR (300 MHz, CDCl 3) δ 2.22 (s, 6H), 2.23 (s, 3H), 4.26 (s, 2H), (s, 2H); 13 C NMR (300 MHz, CDCl3) δ 18.5, 21.3, 43.2, 129.5, 130.4, 135.4, 138.0, 164.8; MS (m/z) (M + ). 2005, Vol. 49, No. 4

5 374 «Á yá xá 3-Bromo-2,4,6-trimethylacetanilide (2) w. y w w. 8 g ye (KBr) 50 ml z, 2 ml(6 g) (Br 2) w w g. w 150 ml v j 2.1 g(10 mmol) yw 1 40 ml 50% k z, ¾ g. yƒ e g. e 5% sodium bisulfite z g 3-bromo-2,4,6-trimethylacetanilide(2) w w. 3-Bromo-2,4,6-trimethylacetanilide (2): mp 168 o C; IR (KBr) 3243, 3100, 3023, 1662, 1581, 1527, 1457, 1419, 1384, 1332, 1234, 1164, 1030 cm 1 ; 1 H NMR (300 MHz, CDCl 3) δ 2.22 (t, 9H), 4.2 6(s, 2H), (s, 1H); MS (m/z) (M + ). N-(3-Bromo-2,4,6-trimethylacetanilido)iminodiacetic acid (3) w. 100 ml ƒ v j 2.9 g(10 mmol) yw 2 š 25 ml k z, 15 ml 1.75 g(9.9 mmol) iminodiacetic acid, disodium salt hydrate. 10% yùp (NaOH) ph 10 j w ƒ y g. w w z, û d l (diethyl ether) 2z w, d ph 2-3 j w e g. e w k z, k w N-(3-bromo-2,4,6- trimethylacetanilido)iminodiacetic acid(3) w w. N-(3-Bromo-2,4,6-trimethylacetanilido)iminodiacetic acid (3): mp 205 o C; IR (KBr) 3447, 3273, 3012, 2964, 1671, 1541, 1378, 1341, 1262 cm 1 ; 1 H NMR (300 MHz, CDCl3) δ (t, 9H), 3.61 (s, 2H), 3.69 (s, 4H), (s, 1 H); 13 C NMR (300 MHz, CDCl 3) δ 17.4, 20.0, 55.7, 58.6, 128.7, 131.4, 135.4, 137.1, 171.9, 173; MS (m/z) (M + ). j q w w ( C). w w Scheme 3 w ùkü. 2,4,6-Trimethylacetanilide (1) w. 30 ml j Scheme 3. Synthesis of N-(3-bromo-2,4,6-trimethylacetanilido) iminodiacetic acid (mebrofenin) using microwave irradiation. q» 2.1 ml(15 mmol) 2,4,6-trimethylaniline š 15 ml p z, 1.37 ml(17 mmol) chloroacetyl chloride. j q e š 300 W w. B w 2,4,6- trimethylacetanilide(1) w w. 2,4,6-Trimethylacetanilide (1): mp 184 o C; IR(KBr) 3236, 3038, 1671, 1608, 1540, 1485, 1333, 1241, 1202, 1039 cm 1 ; 1 H NMR (300 MHz, CDCl3) δ 2.22 (s, 6H), 2.23 (s, 3H), 4.26 (s, 2H), (s, 2H); 13 C NMR (300 MHz, CDCl 3) δ 18.5, 21.3, 43.2, 129.5, 130.4, 135.4, 138.0, 164.8; MS (m/z) (M + ). 3-Bromo-2,4,6-trimethylacetanilide (2) w. 30 ml j q» 1.05 g(5 mmol) yw 1 20 ml 50% k z, B w w. j q e 300 W w. B w 3- Journal of the Korean Chemical Society

6 2005, Vol. 49, No. 4 j q w bromo-2,4,6-trimethylacetanilide(2) w w. 3-Bromo-2,4,6-trimethylacetanilide (2): mp 168 o C; IR (KBr) 3243, 3100, 3023, 1662, 1581, 1527, 1457, 1419, 1384, 1332, 1234, 1164, 1030 cm 1 ; 1 H NMR (300 MHz, CDCl3) δ 2.22 (t, 9H), 4.2 6(s, 2H), (s, 1H); MS (m/z) (M + ). N-(3-Bromo-2,4,6-trimethylacetanilido) iminodiacetic acid (3) w. 30 ml j q» 1.45 g (5 mmol) yw 2 š 13 ml k z, 7 ml 8.8 g(5 mmol) iminodiacetic acid, disodium salt hydrate B w 10% yùp. j q e 300 W w. B w N-(3-bromo- 2,4,6-trimethylacetanilido)iminodiacetic acid(3) w w. N-(3-Bromo-2,4,6-trimethylacetanilido)iminodiacetic acid (3): mp 205 o C; IR (KBr) 3447, 3273, 3012, 2964, 1671, 1541, 1378, 1341, 1262 cm 1 ; 1 H NMR (300 MHz, CDCl 3) δ (t, 9H), 3.61 (s, 2H), 3.69 (s, 4H), (s, 1 H); 13 C NMR (300 MHz, CDCl3) δ 17.4, 20.0, 55.7, 58.6, 128.7, 131.4, 135.4, 137.1, 171.9, 173; MS (m/z) (M + ). Tc-Mebrofenin (4). 10 ml ³ j q w w 1 mg N-(3-bromo- 2,4,6-trimethylacetanilido)iminodiacetic acid(3) š 0.2 ml 3 N HCl z, 3 N HCl 10 µg SnCl 2 2H 2O yww. v 0.9 ml w z, w 0.1 ml(5 mci, 185 MBq) Na TcO 4 š 30 g. ph y t» w ph ƒƒ w ww. t t z, y z ƒ w Tc, š lj p (pertechnetate) ƒ ƒ w» w d j m v(itlc) ww. sy yùp acetonitrile:water(3:1, v/v) yw ƒƒ w z, ITLC-SA ITLC-SG t yw ƒƒ w w. Tc-Mebrofenin w 375 j q w r w w z w y w. j q w lj p- r t w t 99.5% t z ùkü. w» (organ imaging) t w e w p w wƒ v», w w y j q w w w. w j q w» y w w w œw š». x 1. Kim, M. K.; Seidel, J.; Le, N.; Kim, I. S.; Yoo, T. M.; Barker, C.; Kobayashi, H.; Green, M. V.; Carrasquillo, J. A.; Paik, C. H. Nucl. Med. Biol. 1999, 26, Brboric, J. S.; Vladimirov, S.; Jovanovic, M. S.; Dogovic, N. Monatshefte fur Chemie. 2004, 135, Callery, P. S.; Faith, W. C.; Loberg, M. D.; Fields, A. T.; Harvey, E. B. J. Med. Chem. 1976, 19, Nunn, A. D.; Loberg, M.D.; Conley, R. A. J. Nucl. Med. 1983, 24, Brown, P. H.; Juni, J. E.; Liebermann, D. A.; Krishnamurthy, G. T. J. Nucl. Med. 1988, 29, Dogan, A. S.; Conway, J. J.; Lloyd-Still, J. D. J. Nucl. Med. 1994, 35, Gambhir, S. S.; Hawkins, R. A.; Huang, S. C.; Hall, T. R.; Busuttil, R. W.; Phelps, M. E. J. Nucl. Med. 1989, 30, Johnson, K.; Alton, H. M.; Chapman, S. Pediatr. Radiol. 1998, 28, Kuhnert, N. Angew. Chem. Int. Ed. Engl. 2002, 41, Kaboudin, B.; Norouzi, H. Tetrahedron Lett. 2004, 45, Bose, A. K.; Ganguly, S. N.; Manhas, M. S.; Srirajan, V.; Bhattacharjee, A.; Rumthao, S.; Sharma, A. H. Tetrahedron Lett. 2004, 45,

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