PROGRESS IN BIOMEDICAL OPTICS

SERIES

Proceedings of

Photodynamic Therapy of Cancer II

Daniel Brault Giulio Jori Johan Moan Benjamin Ehrenberg Chairs/Editors

Abraham Katzir Biomedical Optics Series Editor

9-10 September 1994 Lille, France

Sponsored by Tie Commission of the European Communities, Directorate General for Science,

Research, and Development INSERM—Institut National de la Sante et de la Recherche Medicale Tie Biomedical Optics Society ELA—The European Laser Association EOS—The European Optical Society S8LM—La Societe Beige de Laser Medical SFLM—La Societe Frangaise des Lasers Medicaux SPIE—The International Society for Optical Engineering

Published by SPIE—The International Society for Optical Engineering Volume 2325

SPIE (The Soc ie ty o f Pho to -Op t i ca l I ns t rumen ta t i on Engineers) is a n o n p r o f i t soc ie ty d e d i c a t e d to t he a d v a n c e m e n t o f op t i ca l and o p t o e l e c t r o n i c app l i ed sc ience and t e c h n o l o g y .

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Please use the fo l lowing format to cite material from this book: Author(s), "Title of paper/' in Photodynamic Therapy of Cancer II, Daniel Brault, G iu l io

Jori, Johan Moan, Benjamin Ehrenberg, Editors, Proc. SPIE 2325, page numbers (1995).

Library of Congress Catalog Card No. 94-67401 ISBN 0-8194-1658-4

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38 Μ t m . Nr.

Contents

ix Conference Committee

S E S S I O N 1 C H E M I S T R Y O F P H O T O S E N S I T I Z E R S

2 Synthet ic a p p r o a c h e s to long-wavelength photosensi t izers for photodynamic therapy a n d the i r p r e l i m i n a r y b io logical act ivity [ 2 3 2 5 - 0 1 ] R. K. P a n d e y , Roswe l l Park Cancer Inst i tu te (USA) ; N . Jagerov ic , U n i v . o f C a l i f o r n i a / D a v i s (USA) ; T. J. D o u g h e r t y , Roswe l l Park Cancer Inst i tute (USA) ; Κ. M . S m i t h , U n i v . o f C a l i f o r n i a / D a v i s (USA)

13 In v i t ro photobio logica l activity of a new ser ies of photosensi t izers: the g lycoconjugated p o r p h y r i n s [ 2 3 2 5 - 0 2 ] M . M o m e n t e a u , D. O u l m i , P. M a i l l a r d , A . Cro isy , Inst i tu t Cu r i e (France)

2 4 T e t r a b e n z o p o r p h y r i n s for P D T [ 2 3 2 5 - 0 3 ] M a . A . V a l l e s , A . M a . G o m e z , U n i v . de Barce lona (Spain)

2 9 Se lec t ive synthes is and photophysica l propert ies of ta i lor-made ch lor ins for p h o t o d y n a m i c therapy [ 2 3 2 5 - 0 4 ] F.-P. M o n t f o r t s , B. C e r l a c h , G . Haake , F. H ö p e r , D. Kusch , A . M e i e r , G . S c h e u r i c h , U n i v . B r e m e n (FRG); H . - D . Brauer, Κ. S c h i w o n , G . S c h e r m a n n , U n i v . Frankfur t (FRG)

4 0 C h l o r i n - t y p e photosensi t i zers der ived f rom vinyl porphyr ins [ 2 3 2 5 - 0 5 ] D. Brau l t , M u s e u m N a t i o n a l d ' H i s t o i r e Na tu re l l e (France); B. A v e l i n e , O . D e l g a d o , C. V e v e r - B i z e t , INSERM (France)

4 7 Intra- a n d in te rmembrane distr ibution of chfor in e6 der ivat ives [ 2 3 2 5 - 0 6 ] V . P. Z o r i n , T. E. Z o r i n a , I. S. M i k h a l o v s k y , I. I. K h l u d e y e v , Bye loruss ian State U n i v .

58 Photomodi f ica t ion of A L A - i n d u c e d protoporphyr in IX in cel ls in vitro [ 2 3 2 5 - 6 2 ] G . S t recky te , V i l n i u s U n i v . (L i thuan ia) ; K. Berg, J. M o a n , Inst i tute for Cance r Research ( N o r w a y )

S E S S I O N 2 P H Y S I C A L C H E M I S T R Y A N D S P E C T R O S C O P Y O F P H O T O S E N S I T I Z E R S

6 8 S p e c t r o s c o p i c studies of photosensit izat ion in solut ions and in cel ls [ 2 3 2 5 - 0 8 ] B. E h r e n b e r g , L. R o i t m a n , A . Lav i , Y. N i t z a n , Z . M a l i k , Bar-Man U n i v . (Israel); J. L. Sessler, U n i v . o f T e x a s / A u s t i n (USA)

8 0 P h o t o p h y s i c a l charac te r i za t ion of far-red absorbing photosensi t izers in cyc lodex t r in so lut ions [ 2 3 2 5 - 0 9 ] B. Röder , C. Z i m m e r m a n n , R. Her ter , H u m b o l d t U n i v . Ber l in (FRG)

92 C a r r i e r sys tems in P D T : on the way to novel ant i tumor drugs [ 2 3 2 5 - 1 0 ] J. G . M o s e r , Α . H e u e r m a n n , P. O e h r , H e i n r i c h - H e i n e - U n i v . (FRG); H . Scheer , L u d w i g - M a x i m i l i a n s - U n i v . (FRG); A . Ve rvoo r t s , S. And rees , H e i n r i c h - H e i n e - U n i v . (FRG)

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1 0 0 Photodynamic therapy of human skin tumors using topical appl icat ion of 5 -aminolevul in ic a c i d , d imethylsul foxide ( D M S O ) , and edet ic ac id d isodium salt (EDTA) [ 2 3 2 5 - 7 2 ] A . O r e n s t e i n , G . Kos ten i ch , Η . Tsur, Sheba M e d i c a l Ctr. (Israel); L. R o i t m a n , B. Eh renberg , Ζ . M a l i k , Bar-Man U n i v . (Israel)

106 Compar a t i ve in vitro and in vivo measurements of hydrophi l ic and hydrophob ic porphyr ins [ 2 3 2 5 - 1 2 ] H . S c h n e c k e n b u r g e r , U n i v . U l m and Fachhochschu le A a l e n (FRG); R. Sai ler, Μ . H . G s c h w e n d , Κ. Kunz i -Rapp , A . C. Rück, W . S. L. Strauß, U n i v . U l m (FRG)

116 T ime- reso lved detect ion of singlet oxygen luminescence in red ce l l ghosts generated by photosensi t izers v ia excitat ion in the far red [ 2325-1 3] S. O e l c k e r s , T. H a n k e , H u m b o l d t U n i v . Ber l in (FRG); J. G . Moser , H e i n r i c h - H e i n e - U n i v . (FRG); B. Röder , H u m b o l d t U n i v . Ber l in (FRG)

1 21 H igh-average-power tunable laser system and its long-term per fo rmance for photodynamic therapy [ 2 3 2 5 - 1 4 ] Μ . V . O r t i z , T. D. C o l e m a n , R. M . Pon , Laserscope Surg ica l Systems (USA)

1 33 Two-step excitat ion in photosensi t ized tumor therapy [ 2325-1 5] R. Ro tomsk is , V . M i c k ü n a i t i s , D. Juodzev i c i us , A . S. Piskarskas, V i l n i u s U n i v . (L i thuan ia)

144 W o r k i n g out the early diagnost ics and controls for the c a n c e r t reatment method with the use of photosensi t izer of model ing act ion [ 2 3 2 5 - 1 6 ] V . B. Loschenov , Genera l Physics Inst i tute (Russia); R. W . Steiner, U n i v . U l m (FRG)

S E S S I O N 3 R O U N D T A B L E : N E W P E R S P E C T I V E S IN P H O T O D Y N A M I C T H E R A P Y

1 50 N e w f luorinated photosensi t izers based on tet rak is(hydroxyphenyl )porphyr ins [ 2325 -1 7] R. Bonne t t , S. P. Songca, Q u e e n M a r y and Wes t f i e l d Co l l ege (UK)

1 55 Immune modulat ion using t ransdermal photodynamic therapy [ 2 3 2 5 - 1 8 ] J. G . Levy , R. K. C h o w d h a r y , Q u a d r a Log ic T e c h n o l o g i e s Inc. (Canada) ; L. Ratkay, D. W a t e r f i e l d , U n i v . o f Br i t ish C o l u m b i a (Canada); M . O b o c h i , S. Leong , D. H u n t , A . H . C h a n , Q u a d r a Log ic T e c h n o l o g i e s Inc. (Canada)

1 66 Photodynamic decontaminat ion of blood for t ransfusion [ 2 3 2 5 - 1 9 ] E. Ben-Hur , H . M a r g o l i s - N u n n o , P. G o t t l i e b , S. Lus t i gman , B. H o r o w i t z , N e w York B l o o d Ctr. (USA)

1 74 Lethal photosensit izat ion of Helicobacter species [ 2 3 2 5 - 2 0 ] C. E. M i l l s o n , U n i v . Co l l ege L o n d o n M e d i c a l Schoo l (UK) ; M . W i l s o n , Eastman Den ta l Ins t i tu te fo r O r a l Hea l th Care Sciences (UK); A . J. MacRobe r t , U n i v . Co l l ege L o n d o n M e d i c a l S c h o o l (UK) ; W . T h u r r e l l , W h i t t i n g t o n Hosp i ta l (UK); P. M l k v y , C. Dav ies , S. G . B o w n , U n i v . C o l l e g e L o n d o n M e d i c a l Schoo l (UK)

S E S S I O N 4 P H O T O D Y N A M I C T H E R A P Y

182 Pr imary targets in photochemica l inactivat ion of cel ls in cul ture [ 2 3 2 5 - 2 1 ] K. Berg, S. G . Jones, Inst i tute for Cancer Research ( N o r w a y ) ; K. Prydz , U n i v . o f O s l o ( N o r w a y ) ; J. M o a n , Inst i tu te for Cancer Research ( N o r w a y )

iv I SPIE Vol. 2325 Photodynamic Therapy of Cancer II (1994)

189 K inet ics of ce l lu la r uptake and retention of the benzoporphyr in der ivat ive (BPD) : r e levance to photodynamic therapy [ 2 3 2 5 - 2 2 ] A . M . Richter , Q u a d r a Log ic T e c h n o l o g i e s Inc. (Canada); H . M e a d o w s , A . K. Jain, A . J. C a n a a n , U n i v . o f Br i t ish C o l u m b i a (Canada); J. G . Levy, Q u a d r a Logic T e c h n o l o g i e s Inc. (Canada)

198 C o m p a r i s o n of the ce l l photodynamic sensitivity for the a luminum phtha locyan ine of dif ferent int racel lu lar loca l izat ions [ 2 3 2 5 - 2 3 ] Ε. B. C h e r n y a e v a , M . Y u . Porosh ina , M o s c o w State U n i v . (Russia); J. G r e v e , A . M . van L e e u w e n , B. G . de G r o o t h , U n i v . o f T w e n t e (Ne ther lands) ; A . P. Savi tsky, A . N . Bach Inst i tu te o f B iochem is t r y (Russia)

21 2 D i s c r e p a n c y between photodynamic injuries and pheophorb ide A a c c u m u l a t i o n in digestive t issues [ 2 3 2 5 - 2 5 ] S. Evrard , M . K o e n i g , C. D a m g e , J. Marescaux , M . A p r a h a m i a n , H ö p i t a u x Un ive rs i ta i res (France)

2 2 0 Pharmacok ine t ics of the far red absorbing octa-a -buty loxy-z inc phtha locyan ine in Lewis lung c a r c i n o m a bear ing mice [ 2 3 2 5 - 2 6 ] C. Dress ler , Freie U n i v . Ber l in and Lase r -Med i z i n -Zen t rum Ber l in G m b H (FRG); M . S. Isma i l , Freie U n i v . Ber l in (FRG); C. N o w a k , M a x - D e l b r ü c k - C e n t r u m (FRG); R. Her ter , T e c h n i s c h e F a c h h o c h s c h u l e Ber l in and H u m b o l t U n i v . Ber l in (FRG); R. G . Senz, L a s e r - M e d i z i n - Z e n t r u m Ber l in G m b H and Techn i sche Fachhochschu le Ber l in (FRG); R. H a g e m a n n , L a s e r - M e d i z i n - Z e n t r u m Ber l in G m b H (FRG); B. Röder, H u m b o l d t U n i v . Ber l in (FRG); H.-P. Be r l i en , Freie U n i v . Ber l in and Lase r -Med i z i n -Zen t rum Ber l in G m b H (FRG)

2 2 8 Ac t ive immunotherapy of Wa lke r -256 c a r c i n o s a r c o m a by tumor-infi l trating lymphocytes assoc ia ted with photodynamic therapy [ 2 3 2 5 - 2 7 ] V . F. D i m a , C a n t a c u z i n o Inst i tute (Romania) ; V . V a s i l i u , Inst i tute o f A t o m i c Physics (Roman ia ) ; D. Laky, V i c t o r Babes Inst i tute (Romania) ; P. l onescu , M i l i t a r y Cent ra l Hosp i t a l (Roman ia ) ; S. V . D i m a , C a n t a c u z i n o Inst i tute (Romania)

2 4 0 C l in ica l photodynamic therapy of mal ignant neoplasms [ 2325 -32 ] Ε. E. S t ranadko , Ο . K. S k o b e l k i n , G . L i t w i n , T. As t r ak hank i na , State Research Ctr. fo r Laser M e d i c i n e (Russia)

SESSION 5 POSTER SESSION

2 4 8 Q u a n t u m yields of photodegradat ion of protoporphyr in IX in solut ion: luminescent , spect rophotomet r ic , and biological testing of photobleaching [ 2 3 2 5 - 3 4 ] L. Bezde tnaya , Ctr . A lex i s Vau t r i n (France) and Russian State M e d i c a l U n i v . ; N . Z e g h a r i , M . B a r b e r i - H e y o b , J.-L. M e r l i n , Ctr. A lex i s Vau t r i n (France); A . Po tapenko , Russian State M e d i c a l U n i v . ; F. H. G u i l l e m i n , Ctr . A lex i s Vau t r i n (France)

2 5 9 Te t rasu lphonated a luminum phtha locyanine as a pH-sensi t ive probe [ 2 3 2 5 - 3 6 ] Ε. B. C h e r n y a e v a , M . Y u . Porosh ina , L. V . Z h o r i n a , Α . V . Ag ronskaya , A . Y u . C h i k i s h e v , V . A . S c h i n o v , S. Y u . A rdgan tsev , M o s c o w State U n i v . (Russia); M . G . G a l p e r n , M o s c o w Inst i tute o f O r g a n i c In te rmedia tes and Dyes (Russia)

2 7 0 F l u o r e s c e n c e detect ion system using a krypton laser: appl icat ion to c a n c e r diagnosis in photodynamic therapy [ 2 3 2 5 - 3 7 ] M.-L. D i l l e r , Y. G r a n j o n , Ctr. de Recherche en A u t o m a t i q u e de N a n c y (France); F. H . G u i l l e m i n , Ctr. A lex i s Vau t r i n (France); E. Y v r o u d , Ctr. de Recherche en A u t o m a t i q u e de N a n c y (France)

SPIE Vol. 2325 Photodynamic Therapy of Cancer II (1994) Ι ν

2 7 7 C om par a t i ve analys is of different p h t h a l o c y a n i n e photosensi t izers for exper imenta l P D T of c a n c e r [ 2 3 2 5 - 3 9 ] M . G . G a l p e r n , I. N o v o z h i l o v a , G . Sokol»ova, G . N. V o r o z h t s o v , M o s c o w Inst i tu te o f O r g a n i c In te rmed ia tes and Dyes (Russia); N . N . Z !harkova, D. N . K o z l o v , Gene ra l Physics Inst i tu te (Russia); V . V . S o k o l o v , G . M . S u k h i n , P.A. He r t zen M o s c o w Research O n c o l o g y Inst i tu te (Russia)

2 8 1 D y n a m i c s of phtha locyanine AI a c c u m u l a t i o n in s tomach c a n c e r photodynamic therapy [ 2 3 2 5 - 4 0 ] S. S. Kharnas, M o s c o w M e d i c a l Academy/ (Russia); V . B. Loschenov , A . A . S t r a t o n n i k o v , T. A . K r a m a r e n k o , Genera l Physics Ins t i tu te (Russia); Ο . V . A r t e m j e v a , M o s c o w M e d i c a l A c a d e m y (Russia); V . P. Bakon in , Μ . I. K iuz in , V . Ya. Z a v o d n o v , G e n e r a l Physics Ins t i tu te (Russia); R. W . Ste iner , U n i v . U l m (FRG)

2 9 0 Photodynamic therapy of colorecta l c a n c e r using a new light s o u r c e : f rom in vi tro studies to a patient t reatment [ 2 3 2 5 - 4 1 ] H . Kashtan, R. H a d d a d , Y. Skorn ick , T e l - A v i v M e d i c a l Ctr . and Sackler Schoo l o f M e d i c i n e (Israel)

2 9 7 N e w ch lor in and bacter iochlor ine- type (photosensit izers for photodynamic therapy [ 2 3 2 5 - 4 7 ] A . N . Kozy rev , Α . V . E f imov , O . A. Efrermova, P. Y u . P e r e p y o l k i n , A . F. M i r o n o v , M .V . L o m o n o s o v M o s c o w State A c a d e m y o f Fine C h e m i c a l T e c h n o l o g y (Russia)

3 0 6 Combina t ion of photodynamic therapy a n d x- i rradiat ion: a study on 5-aminolevul in ic ac id (ALA) radiomodify ing properties [ 2 3 2 5 - 4 9 ] Z . Luks iene , L i t huan ian O n c o l o g i c a l Ctr.;; K. Berg, J. M o a n , N o r w e g i a n Rad ium H o s p i t a l

3 1 2 Synthesis and propert ies of e ther -bonded porphyr in -ch lor in d imers [ 2 3 2 5 - 5 0 ] A . F. M i r o n o v , E. G . Lev inson , M.V. L o m o n o s o v M o s c o w State A c a d e m y o f Fine C h e m i c a l T e c h n o l o g y (Russia)

321 F l u o r e s c e n c e detect ion of MSH-receptoirs in me lanoma as a target of hormone-d i rec ted photosensi tat ion in P D T [ 2325-51 ] S. Roehrs , j . G . M o s e r , H e i n r i c h - H e i n e - U n i v . (FRG); Y. S a l o m o n , W e i z m a n n Inst i tu te o f Sc ience (Israel)

3 2 6 Photodynamic therapy of human b ladder c a r c i n o m a cel ls in vitro with l iposomes as a c a r r i e r for protoporphyr indisodiumsal t [2325-5 2] K. Schm id t , U . K. W e n d e r o t h , Ε. Reich, R:. Ε. H a u t m a n n , U n i v . U l m (FRG)

3 3 9 S p e c t r o s c o p i c studies on Si-phthalocyamines and S i -naphtha locyanines [ 2 3 2 5 - 5 3 ] C. v o n S c h ö n e r m a r k , A . V o l k m e r , H u m b o l d t U n i v . Ber l in (FRG); S. M ü l l e r , D. W ö h r l e , U n i v . B remen (FRG); B. Röder, H u m b o l d t U n i v . Ber l in (FRG)

3 4 9 Interesting method for the synthesis of rnonofunct ional ized phtha locyan ines v ia subphtha locyan ines for photodynamic tlherapy of c a n c e r [ 2 3 2 5 - 5 4 ] R. G . Senz, T e c h n i s c h e Fachhochschu le Ber l in and Lase r -Med i z i n -Zen t rum Ber l in G m b H (FRG); R. Her ter , T e c h n i s c h e Fachhochschu le Be r l i n (FRG)

351 Mult iple laser i rradiat ion of O R L organ Humors in photodynamic therapy [ 2 3 2 5 - 5 5 ] V . V . Shen ta l , N . E. Edynak, N. A. A b d u l i i n , Y. P. K u v s h i n o v , Β. K. P o d d u b n y i , T. D. T a b o l i n o v s k a y a , O n c o l o g i c a l Sc ien t i f i c Ctr. (Russia); V . B. Loschenov , P. V . P o l e s h k i n , E. A . Luk ' yane ts , Gene ra l Physics Ins t i tu te (Russia)

v/ / SPIE Vol. 2325 Photodynamic Therapy of Cancer II (1994)

355 I n f l u e n c e o f p h o t o d y n a m i c t h e r a p y o n t h e d e l a y o f metastas is d e v e l o p m e n t in Lew is l u n g c a r c i n o m a [ 2 3 2 5 - 5 6 ] V . M a n t a r e v a , Inst i tute o f O r g a n i c Che m i s t r y (Bulgar ia) ; K. Kassabov, N a t i o n a l Ctr . o f O n c o l o g y (Bulgar ia) ; M . Shopova , Inst i tu te o f O r g a n i c Chemis t r y (Bulgar ia) ; S. M ü l l e r , D. W ö h r l e , U n i v . B remen (FRG)

3 6 4 F i rs t c l i n i c a l resu l ts w i t h a n e w d r u g f o r P D T [2325 -57 ] V . V . S o k o l o v , V . I. Ch issov , Ε. V . F i l o n e n k o , R. I. Yakubovskaya , D. G . S u k h i n , P.A. H e r t z e n M o s c o w Research O n c o l o g y Inst i tute (Russia); M . G . G a l p e r n , G . N . V o r o z h t s o v , M o s c o w Research Inst i tu te o f O r g a n i c In termedia tes and Dyes (Russia); Α . V . G u l i n , Μ . B. Z h i t k o v a , Research Inst i tu te Pol jus (Russia); N . N . Z h a r k o v a , D. N . Koz lov , V . V . S m i r n o v , G e n e r a l Phys ics Inst i tu te (Russia)

3 6 7 P h o t o d y n a m i c t h e r a p y o f c a n c e r w i t h t h e p h o t o s e n s i t i z e r P H O T O G E M [ 2 3 2 5 - 5 8 ] V . V . S o k o l o v , V . I. Ch issov , Ε. V . F i l o n e n k o , G . M . S u k h i n , R. I. Y a k u b o v s k a y a , T. A . Be lous , P.A. He r t zen M o s c o w Research O n c o l o g y Inst i tute (Russia); Ν . N . Z h a r k o v a , D. N . K o z l o v , V . V . S m i r n o v , Gene ra l Physics Inst i tute (Russia)

375 C l i n i c a l f l u o r e s c e n c e d iagnos t i cs in t h e c o u r s e o f p h o t o d y n a m i c t h e r a p y o f c a n c e r w i t h t h e p h o t o s e n s i t i z e r P H O T O G E M [ 2 3 2 5 - 5 9 ] V . V . S o k o l o v , V . I. Ch issov , Ε. V . F i l o n e n k o , P.A. Her tzen M o s c o w Research O n c o l o g y Ins t i tu te (Russia); Ν . N . Z h a r k o v a , D. N . K o z l o v , V . V . Sm i rnov , Genera l Physics Inst i tu te (Russia)

381 E x p e r i m e n t a l s tud ies o f t h e c o m b i n a t i o n o f PDT and t u m o r c h e m o t h e r a p y o r 6 0 C o i r r a d i a t i o n [ 2 3 2 5 - 6 0 ] J. D i d z i a p e t r i e n e , G . P rasmick iene , D. Sukeliene, L i thuan ian O n c o l o g y Ctr. ; R. Ro tomsk is , G . S t recky te , V . A t k o c i u s , L. S tac iok iene , V . Smi lgev i c ius , V i l n i u s U n i v . (L i thuan ia)

3 8 9 I n v e s t i g a t i o n o f p h t a l o c y a n i n e A l p h o t o s e n s i t i z e r and b l o o d i n t e r a c t i o n [ 2 3 2 5 - 6 3 ] N . L. T o r s h i n a , V . B. Loschenov , A . A . S t ra tonn ikov , A . Y u . D u p l i k , A . M . Posypanova , G e n e r a l Physics Inst i tu te (Russia)

391 Q u a n t i t a t i v e d a t a o n b l o o d f l o w d u r i n g t u m o r PDT o b t a i n e d by laser D o p p l e r s p e c t r o s c o p y in t h e hen ' s egg tes t sys tem [2325 -65 ] A . V e r v o o r t s , A . R o o d , H e i n r i c h - H e i n e - U n i v . (FRG); M . K lo tz , K lo tz A n a l y t i c a l Regis t ra t ion T e c h n i q u e s (FRG); J. G . Mose r , H e i n r i c h - H e i n e - U n i v . (FRG); M . R o s e n b r u c h , Bayer A G (FRG)

4 0 0 F l u o r e s c e n c e o b s e r v a t i o n s o f pa t i en t s in t h e c o u r s e o f p h o t o d y n a m i c t h e r a p y o f c a n c e r w i t h t h e p h o t o s e n s i t i z e r P H O T O S E N S [ 2 3 2 5 - 6 7 ] Ν . Ν . Z h a r k o v a , D. Ν . K o z l o v , V . V . S m i r n o v , Genera l Physics Inst i tu te (Russia); V . V . S o k o l o v , V . I. Ch i ssov , Ε. V . F i l o n e n k o , D. G . S u k h i n , P.A. Her tzen M o s c o w Research O n c o l o g y Ins t i tu te (Russia); M . G . G a l p e r n , G . N . V o r o z h t s o v , M o s c o w Research Inst i tu te o f O r g a n i c In te rmed ia tes and Dyes (Russia)

4 0 4 L i ke l y m e c h a n i s m o f se lec t i ve p h o t o s e n s i t i z e r a c c u m u l a t i o n in m a l i g n a n t t u m o r s : t h e m a t h e m a t i c a l m o d e l [ 2325 -07 ] Ν . V . S tepanova , L. V . Z h o r i n a , Ε. B. Che rnyaeva , M o s c o w State U n i v . (Russia)

4 1 6 P h e o p h o r b i d e s as p h o t o s e n s i t i z e r s f o r t h e p h o t o d y n a m i c t h e r a p y o f t u m o r s [ 2325 -1 1 ] C. T a n i e l i a n , C. W o l f f , Ecole Eu ropeenne des Hautes Etudes des Indust r ies (France); M . Kobayash i , U n i v . o f T o k y o (Japan)

4 2 5 P h o t o d y n a m i c t h e r a p y o f h u m a n t u b u l o - v i l l o u s a d e n o m a s [ 2 3 2 5 - 7 3 ] T. W a r l o e , Q . Peng, H. H e y e r d a h l , H . Waahre, J. M o a n , H . Steen, K.-E. G i e r c k s k y , N o r w e g i a n Rad ium Hosp i t a l

4 3 7 Author Index

SPIE Vol. 2325 Photodynamic Therapy of Cancer II (1994) f vii

Conference Committee

Conference Chairs

Daniel Brault, Museum National d'Histoire Naturelle (France) Giulio Jori, University of Padova (Italy) Johan Moan, Norwegian Radium Hospital (Norway) Benjamin Ehrenberg, Bar-llan University (Israel)

Session Chairs

1 Chemistry of Photosensitizers Daniel Brault, Museum National d'Histoire Naturelle (France)

2 Physical Chemistry and Spectroscopy of Photosensitizers Benjamin Ehrenberg, Bar-llan University (Israel)

3 Roundtable: New Perspectives in Photodynamic Therapy Giulio Jori, University of Padova (Italy)

4 Photodynamic Therapy Giulio Jori, University of Padova (Italy) Johan Moan, Norwegian Radium Hospital (Norway)

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Carrier systems in PDT: On the way to novel anti-tumor drugs

Jörg G.Moser1, Anja Heuermannl, Peter Oehr 1, Hugo Scheer2, Anja Vervoorts1, and Sonja Andrees1

11nstitute of Laser Medicine, Heinrich-Heine-University, Universitaetsstr.1 D-40225 Duesseldorf (Germany)

2lnstitute of Botany, Ludwig-Maximilians-University, Menzänger Str. 67 D-80638 Munich (Germany)

Abstract

A novel mode to apply photosensitizing drugs specifically to tumor tissue using the principles of polyphasic tumor therapies is lined out. Key compounds are tumor specific functionalized antibodies with reduced immunogenicity. These bind to drug inclusion complexes in a multiplicative manner. Drug inclusion complexes are designed on the basis of tethered functionalized ß-cyclodextrin dimers with maximum affinity to porphyrinoid photosensitizers forming monomeric chlathrates. To enhance porphyrin-cyclodextrin interaction peripheral groups of the porphyrin have to be chemically modified. The development of the method is not yet completed. First results are demonstrated.

Introduction

In a tumor-bearing organism there are two general routes by which photosensitizers find their target tissue: water soluble sensitizers (e.g. the polysulphonated m-tetraphenyl porphines or phthalocyanines) enrich in tumor tissue according to pH-equilibria of their differently ionized forms (1). Apolar photosensitizers use the lipoprotein pathway to be taken up as complexes via the lipoprotein receptors of the tumor cells (2). Both these pathways allow for targeting of healthy organs and lead to the wellknown side effects of PDT as well as of general chemotherapy.

To avoid these side effects we started with the simple consideration to construct a porphyrinoid -containing inert drug which cannot be taken up by any living cell. Such a drug should be functionalized in a manner that it can be bound exclusively to tumor cells. The stability of such a drug should be sufficiently high to prevent interaction with the known distribution pathways.

Water soluble complexes: cyclodextrins

Cyclodextrins are wellknown to form soluble complexes with several water-insoluble drugs (3). For example, α-cyclodextrin is admitted as an aid to solubilize prostaglandin Ε 1 for injections, γ-cyclodextrin has been used successfully to solubilize tin etio-purpurin for PDT purposes (4). In general, aggregates of apolar photosensitizers dissolve in cyclodextrin solutions into monomeric host-guest complexes (fig. 1). Highly symmetrical apolar phthalocyanins and porphycenes achieve an orientation towards the substructures of the cell membrane, and this results in a greatly enhanced uptake velocity by the cell (fig. 2). Despite the considerable stability of the 1:1 host-guest complexes (fig. 3) an interaction with the lipoprotein pathway is pre-programmed (4).

92 /SPIE Vol. 2325 Photodynamic Therapy of Cancer II (1994) 0-8194-1658-4/95/$6.00

7 0 0 λ [nm] 8 0 0

1/[CO|l/mol -12000.00 -3000.00 -4000.00 0.00 4000.00 8000.00 1/1CO] l/mol

fig. 3 Association equilibria of tetra-t-butyl-phenoxyphthalocyanine and BPEE as revealed by TNS-exclusion fluorescence technique. Kp (TTBP) = 14.8 χ 10"6M

K D(BPEE) = 8.3x10' 6 M

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Stabilization of the complex: cyclodextrin oligomers

As a signpost towards stable complexes, BRESLOW and CHUNG (5) investigated the complexation of a very simple symmetrical diester, propane diol p-tert.butyl benzoic acid diester, with a dimeric "clamshell" ß-cyclodextrin. The stability of the complex exceeded the expected value by two orders of magnitude and resulted in a KQ = 10~ 1 0 M comparable with the complex stability between antibodies and their antigens. In the mean time several other groups constructed cyclodextrin dimers mainly to study and to mimick enzyme-like activities. Our first model of a more flexible cyclodextrin dimer to complex porphyrinoid compounds (6) has been reconstructed to a box-of-bricks principle (fig. 4): starting from tosylated ß-cyclodextrin (at positions 2 or 6) we substitute by nucleophilic attack with alkyl diamines of short chain length and connect the resulting N-alkyl amines with homobifunctional reagents to form ß-cyciodextrin dimers with variable extension including the possibility to introduce preformed break-points to finally break the complex at the target site.

Fx

fig. 4 Orientation of CD's against guests and bridge forming modalities to achieve optimum binding

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High affinity complexes: anchor structures

Porphyrins normally contain very short side chains which allow only poor interactions with the extended lipophilic cavity of ß-cyclodextrin. More voluminous side chains, like the p-tert.butyl phenyl group, may allow for much higher stability of the host-guest complex, and by the same time may enhance the dipole momentum of the included drug (7). At the moment we try to substitute pheophorbide a in C-31 with a p-tert.butyl phenyl ether according to the general method given by PANDEY et al (8) in order to better understand anchor functions (fig.5).

fig. 5 Substitution of apolar side chains may enhance complex stability

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Functionalization of the complex: biotin-avidin complexes

The biotin-avidin system is wellknown to improve considerably the tumor localization by radioscintigraphic methods (9).Thus, biotinylation of the drug-cyclodextrin complexes as well as of monoclonal tumor-localizing antibodies is investigated in our group. An anti-CEA antibody, B W 431/26, has been biotinylated successfully with up to 7 biotin molecules without loosing its affinity to the antigen localized on the surface of LoVo adenocarcinoma cells. Biotinylation of ß-cyclodextrins in position 6 over a Cg-spacer was easily achieved using the methods yet described.

Immunological problems: immunogenicity of foreign proteins

The introduction of foreign proteins, avidin from hen's egg white and mouse monoclonal antibodies, into the system raises immunological problems. BOSSLET et al.(10) circumwent the problem by constructing an ingenious fusion protein to be applied with the Antibody Directed Enzyme Prodrug Therapy (ADEPT) splitting an adriamycin glucuronide at the target site by a humanized antibody-ß-glucuronidase complex. This is beyond our today facilities. So we study the reduction of immunogenicity by coupling our proteins with unifunctional m-polyethylene glycols (11) which in a modified form may be used as additional centers to accumulate the drug complexes.

fig. 6 High stability of inclusion complexes prevents sensitizer transport on the lipoprotein pathway

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Outline of the targeting system: polyphasic PDT

The intended targeting approach is closely related to the ADEPT concept just in phase II-studies in Great Britain and in Germany. In a first step, biotinylated antibodies with tumor-localizing properties are injected and then cleared for some days as controlled by scintigraphy. In a second step avidin or just avidin-sensitizer chlathrate complexes can be injected at very low concentrations due to the high affinity to biotin. We expect concentrations as low as 10~1 2M to be able to saturate the antibody-bound biotins excluding any toxicity of ß-cyclodextrin. The multiplicative effect due to multiple binding sites of avidin and the polyvalency of the drug complexes should allow for an accumulation of 10 6 to 10 s molecules of sensitizer per tumor cell assuming 5 χ 10 4 antigenic binding sites. This is fairly in the range of phototoxic concentrations and should allow for irreversible damage by laser irradiation (fig. 6).

fig. 7 Sequential administration of antibodies and drug complexes leads to accumulation of sensitizer due to polyvalency of the single compounds

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A glance over the plate's border: tumor chemotherapy

We should take in mind that complexation of drugs by cyclodextrins is originally invented to solubilize water-insoluble drugs in general. Porphyrinoid binding is only one of the possible applications. Looking over the manifold of today anti-cancer agents there are several potent but harmful drugs which could be detoxified by inclusion into cyclodextrin oligomers and, by chance, transferred to its target without considerable side effects. Moreover, there are several potent anti-cancer drugs which cannot be given to a patient because of their high general toxicity. Some of them are porphyrin derivatives. So, in future such a system may be helpful to revive and to modify general chemotherapy to a curative modality without its inherent side effects.

Literature cited

(1) Pottier R, Kennedy JC (1990): The possible role of ionic species in selective biodistribution of photochemotherapeutic agents toward neoplastic tissue. J. Photochem. Photobiol. 8, 1-16

(2) Hamblin MR, Newmann EL (1994): On the mechanism of the tumor-localizing effect in photodynamic therapy. J. Photochem. Photobiol. 23, 3-8

Jori G, Reddi Ε (1993): The role of lipoproteins in the delivery of tumour-targeting photosensitizers. Int.J.Biochem. 25, 1369-1375

(3) Pitha J (1989): Cyclodextrins: solutions to insolubility. Neurotransmissions 5, (1) 1-4

(4) Kessel D, Morgan A, Garbo GM (1991): Sites and efficency of photodamage by tin etiopurpurin in vitro using different delivery systems. Photochem. Photobiol. 54, 193-196

(5) Breslow R., Chung S (1990): Strong binding of ditopic substrates by a doubly linked occlusive C^'clamshell" as distinguished from an aversive C 2 "loveseat" cyclodextrin. J.Am.Chem.Soc. 112, 9659-9660

(6) Moser JG t Haller C.,Herchenbach B, Vervoorts A (1991): How to prevent loss of photosensitizer into non-cancer tissues? Abstr. 4th Congr.ESP, Amsterdam. A 12

Moser JG, Vervoorts A, Billen A (1992): Molecular encapsulation of pheophorbides as chlathrates in dimeric cyclodextrins. Photodynamic Therapy and Biomedical Lasers (Spinelli ed.) 760-763

(7) Schnurpfeil G.(1994) unpublished results (8) Pandey RK, Shian FY, Sumlin AB, Dougherty TJ, Smith KM (1992): Structure/activity

relationship among photosensitizers related to pheophorbides and bacteriopheophorbides. Bio.Medicinal Chem.Lett. 2, 491-496

(9) Oehr P.,Liu Q, Schultes B, Altreuther M, Pollok J, Biersack HJ (1990): Effect of streptavidin on the localization of tumors by 99m-Tc-NHS-biotin. Conf.Proc. Bad GasteinVol. 19

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(10) Bosslet Κ, Czech J, Lorenz Ρ, Sedlacek HH, Schuermann M, Seemann G.(1992): Molecular and functional characterization of a fusion protein suited for tumour specific prodrug activation. Br.J.Cancer 65, 234-238

Bagshawe KD (1987): Antibody directed enzymes revive anticancer prodrugs concept. Br J.Cancer 56, 531

(11) Abuchowski A, Van Es Τ, Palczuk NC, Davis FF (1977): Alteration of immunological properties of bovine serum albumin by covalent attachment of polyethylene glycol. J.BioL Chem. 252, 3578-3581

Acknowledgements

This work is granted by a fellowship of the Ministry for Research and Technology (BMFT) of the Federal Republic of Germany. The fruitful cooperation with all groups of the cooperation "Photodynamische Tumortherapie", in particular the groups of B. Roeder (Berlin), D. Wöhrle (Bremen), Α Rück (Ulm), and Ε. Vogel (Köln) is thankfully acknowledged.

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