Synthesis, Spectroscopy, and Photocytotoxicity of Glycosylated Amino Acid Porphyrin Derivatives as Promising Molecules for Cancer Phototherapy

Article abstract of DOI:10.1021/jo982499+

To obtain molecules that can target malignant cells, two series of new meso glucosylporphyrins bearing amino acid residues are synthesized in four steps. The first series contained n meso glycosyl moieties and (4 -n) alanyl groups on the ortho or para positions of the meso phenyl. In the second series, the carbohydrate moiety is separated from the aryl substituent by a serine unit. Starting from p- or o-nitrobenzaldehyde, p- or o-acetylbenzaldehyde or -tolualdehyde, and pyrrole, the glycosylnitrophenylporphyrins 3-6 and tritolylporphyrins 8a,b are synthesized under optimized conditions tailored from Lindsey's method. The nitro function is then reduced and N-Fmoc-L-alanine or acetylglycosylated N-Fmoc-serine are coupled on the amino function. A detailed ¹H and ¹³C NMR study allows complete structural elucidation. The UV-visible fluorescence and MALDI mass spectra are presented. Compounds 19-22 produced ¹O₂, and photocytotoxicities against the K562 leukemia cell line are compared to hematoporphyrin. As a result of their sensitizing abilities, these resultant compounds are of considerable interest for photodynamic therapy.

Full text of DOI:10.1021/jo982499+

J . Org. Chem. 1999, 64, 4431-4444
4431
Syn th esis, Sp ectr oscop y, a n d P h otocytotoxicity of Glycosyla ted
Am in o Acid P or p h yr in Der iva tives a s P r om isin g Molecu les for
Ca n cer P h ototh er a p y
V. Sol, J . C. Blais, V. Carr e´ , R. Granet, M. Guilloton, M. Spiro, and P. Krausz*,†
†
‡
§
†
§
†,|
Universit e´ de Limoges-Laboratoire de Chimie des Substances Naturelles 123, Avenue Albert Thomas,
7060 Limoges, France, Universit e´ Pierre et Marie Curie, Laboratoire de Chimie Organique Structurale et
Biologique, Centre National de la Recherche Scientifique EP 1034, Place J ussieu, 75005 Paris, France,
and Universit e´ de Limoges-Institut de Biotechnologie 123, Avenue Albert Thomas,
8
8
7060 Limoges, France
Received December 23, 1998
To obtain molecules that can target malignant cells, two series of new meso glucosylporphyrins
bearing amino acid residues are synthesized in four steps. The first series contained n meso glycosyl
moieties and (4 - n) alanyl groups on the ortho or para positions of the meso phenyl. In the second
series, the carbohydrate moiety is separated from the aryl substituent by a serine unit. Starting
from p- or o-nitrobenzaldehyde, p- or o-acetylbenzaldehyde or -tolualdehyde, and pyrrole, the
glycosylnitrophenylporphyrins 3-6 and tritolylporphyrins 8a ,b are synthesized under optimized
conditions tailored from Lindsey’s method. The nitro function is then reduced and N-Fmoc-L-alanine
1
13
or acetylglycosylated N-Fmoc-serine are coupled on the amino function. A detailed H and C NMR
study allows complete structural elucidation. The UV-visible fluorescence and MALDI mass spectra
1
are presented. Compounds 19-22 produced O
2
, and photocytotoxicities against the K562 leukemia
cell line are compared to hematoporphyrin. As a result of their sensitizing abilities, these resultant
compounds are of considerable interest for photodynamic therapy.
The chemistry of porphyrins bearing glycosylated
groups has made rapid progress during recent years
owing to the development of new photosensitizers being
applied in cancer photochemotherapy.1 Photodynamic
therapy (PDT) is based on the principle that porphyrins
become concentrated in tumor cells and, upon subsequent
irradiation with visible light in the presence of oxygen,
It occurred to us that porphyrins linked to a cellular
recognition element and an hydrophilic moiety could
appear as promising candidates for an application in
PDT. Thus, porphyrins with sugar moieties have not only
good solubility in aqueous solutions but also possible
specific membrane interaction.3 This property can be
enhanced by use of appropriately selected short oligopep-
tides. Indeed, it was recently shown that two tripeptides
2
specifically destroy the cells. Although the exact mech-
(Arg-Gly-Asp and Asn-Gly-Arg) linked to a chemothera-
anism of PDT in cancer treatment is unknown, there is
scope for exploring new variations to amplify the thera-
peutic efficiency of the senzitizer.
peutic molecule (doxorubicin) targets the drug to the new
blood vessels that nourish the tumor.4 This demon-
strates that attaching small peptides to cancer drugs can
b,c
4
enhance their efficacy, reduce their toxicity, and also
*
To whom correspondence may be adressed. Fax: 33.(0)5.55.45.72.02.
improve tumor cell targeting. Moreover, as glycopeptides
and glycoproteins have been involved in important
biological recognition phenomena and transport pro-
E.mail: krausz@unilim.fr.
†
Universit e´ de Limoges-Laboratoire de Chimie des Substances
Naturelles.
‡
Universit e´ Pierre et Marie Curie.
Universit e´ de Limoges-Institut de Biotechnologie.
Permanent address: University of Tirana, Laboratory of Biophys-
5
§
cesses, they have been used as carriers of metabolite
|
inhibitors and toxic drugs; from this perspective and as
a first approach, we have synthesized (i) mono-, di-, and
tri-alanine glycoporphyrin derivatives 19a ,b, 20, 21, and
22a ,b (Scheme 1). Indeed, the free R-amino group of
alanine is known to improve specific cancer cells
targeting.4 In a second step, (ii) two tristolylporphyrins
are substituted by a glycosylated serine 23a ,b because
the presence of lipophilic aryl groups and hydrophilic
substituents could increase the interaction with the
ics, Faculty of Natural Sciences, Tirana, Albania.
1) (a) Czuchajowski, L.; Li, H. Tetrahedron Lett. 1994, 35, 1629.
b) Cornia, M.; Casiraghi, G.; Binacchi, S.; Zanardi, F.; Rassu, G. J .
(
(
Org. Chem. 1994, 59, 1226. (c) Momenteau, M.; Oulmi, D.; Maillard,
Ph.; Croisy, A. SPIE-Int. Soc. Opt. Eng. 1994, 13, 2325. (d) Oulmi, D.;
Maillard, Ph.; Guerquin-Kern, J . L.; Heul, C.; Momenteau, M. J . Org.
Chem. 1995, 60, 1554 and reference therein. (e) Gaud, O.; Granet, R.;
Kaouadji, M.; Krausz, P.; Blais, J . C.; Bolbach, G. Can. J . Chem. 1996,
a
7
4, 481. (f) Driaf, K.; Granet, R.; Krausz, P.; Kaouadji, M.; Thomasson,
F.; Chulia, A. J .; Verneuil, B.; Spiro, M.; Blais, J . C.; Bolbach, G. Can.
J . Chem. 1996 74, 1550. (g) Hombrecher, H. K.; Ohm, S.; Koll, D.
Tetrahedron 1996, 52, 5441. (h) Hombrecher, H. K.; Schell, C.; Thiem,
J . Bioorg. Med. Chem. Lett. 1996, 6, 1199. (i) Sol, V.; Blais, J . C.;
Bolbach, G.; Carr e´ , V.; Granet, R.; Guilloton, M.; Spiro, M.; Krausz,
P. Tetrahedron Lett. 1997, 38, 6391. Mikata, Y.; Onchi, Y.; Tabata,
K.; Ogura, S.; Okura, I.; Ono, H.; Yano, S. Tetrahedron Lett. 1998, 39,
(3) (a) Kieda, C.; Monsigny, M. Invasion Metastasis 1986, 6, 347.
(b) Monsigny, M.; Roche, A. C.; Midoux, P.; Kieda, C.; Mayer, R. Lectins
and glycoconjugates in oncology: structure, function, clinical applica-
tion; Gabius, H. J ., Nagel, G. A., Eds.; Springer-Verlag: Heidelberg,
1988; 1.
4
505.
2) (a) Dougherty, T. J .; Grindey, G. B.; Fiel, R.; Weishaupt, K. R.;
(
Boyle, D. G. J . Natl. Cancer Inst. 1975, 55, 115. (b) Kessel, D. Biochem.
Photobiol. 1986, 43, 681. (c) Van der Berg, H. Chem. Br. 1986, 22, 430.
(4) (a) Tsay, B. L.; Wolfinbarger, L., J r. Cancer Chemother. Phar-
macol. 1987, 19, 190. (b) Barinaga, M. Science 1998, 279, 323. (c) Arap,
W.; Pasqualini, R.; Ruoslahti, E. Science 1998, 279, 377.
(5) Kunz, H. Pure Appl. Chem. 1993, 65, 1223.
(
d) Henderson, B. W.; Dougherty, T. J . Photochem. Photobiol. 1992,
5, 145.
5
1
0.1021/jo982499+ CCC: $18.00 © 1999 American Chemical Society
Published on Web 05/15/1999

4
432 J . Org. Chem., Vol. 64, No. 12, 1999
Sol et al.
Sch em e 1a
a
a and b refer to ortho and para position, respectively.
lipidic membrane. Several examples of coupling of amino
acids to porphyrins have been reported in the literature,
the great majority of which have been designed as models
Resu lts a n d Discu ssion
Syn th esis. The different porphyrins were prepared
according to the general route outlined in Schemes 2 and
3. Aldehyde 1a was prepared from commercially available
helicin (2-(â-D-glucopyranosyloxy)benzaldehyde) by acetyl-
ation at 0 °C during 50 min with acetic anhydride in dry
pyridine (yield 80%). 4-(2′,3′,4′,6′-tetra-O-acetyl-â-D-glu-
copyranosyloxy)benzaldehyde 1b was synthesized from
6
of hematoproteins or for studies in electron-transfer
7
reactions. They have been also used in magnetic reso-
8
9
nance imaging and in DNA sequence recognition.
In the present paper, we report full experimental data
concerning the synthesis and characterization of amino
acid glycosylporphyrin derivatives 19a ,b, 20, 21, 22a ,b,
and 23a ,b (Scheme 1) and compare their in vitro photo-
cytotoxic activities with hematoporphyrin (HP).
2
4
,3,4,6-tetra-O-acetyl-R-D-glucopyranosyl bromide and
-hydroxybenzaldehyde (3 equiv) in acetone in the pres-
1
0
ence of 7% aqueous NaOH. After 24 h, the expected
compound was obtained in 35% yield. Compound 2
(
6) (a) Smith, K. M.; Milgrom, L. R. J . Chem. Soc., Perkins Trans.
1981, 2065 and references therein. (b) Nishino, N.; Mihara, H.;
Hasegawa, R.; Yanai, T.; Fujimoto, T. J . Chem. Soc., Chem. Commun.
992, 692. (c) Akerfeldt, K. S.; Kim, K. M.; Canac, D.; Groves, J . T.;
(Scheme 3) was prepared from â-D-glucose peracetate and
1
N-Fmoc-L-serine with BF O (3 equiv) in acetonitrile
3
‚Et
2
11
1
for 2 h at room temperature. Workup gave compound
Lear, J . D.; Degrado, W. F. J . Am. Chem. Soc. 1992, 114, 9456. (d)
Choma, C. T.; Kaestle, K.; Akerfeldt, K. S.; Kim, R. M.; Groves, J . T.;
DeGrado, W. F. Tetrahedron Lett. 1994, 35, 6191.
2 in a 30% yield. Nitrophenylporphyrin derivatives 3-8
12
were synthesized following Lindsey’s method. This
(7) (a) Pispise, B.; Venanzi, M.; Palleschi, A.; Zanotti, G. Macromol-
ecules 1994, 27, 7800. (b) Hayashi, T.; Takimura, T.; Ohara, T.; Hitomi,
Y.; Ogoshi, H. J . Chem. Soc., Chem. Commun. 1995, 2503.
(10) (a) J eanloz, R. W.; Stoffyn, P. J . Methods Carbohydr. Chem.
1962, 1, 221. (b) Schuster, B.; Winter, M.; Hermann, K. Z. Naturforsh.
1986, 41, 511.
(11) Steffan, W.; Schutkowski, M.; Fischer, G. Chem. Commun.
1996, 313 and references therein.
(12) Lindsey, J . S.; Schreiman, I. C.; Hsu, H. C.; Kearney, P. C.;
Marguerettaz, A. M. J . Org. Chem. 1987, 52, 827.
(
8) Matthews, S. E.; Pouton, C. W.; Threadgill, M. D. J . Chem. Soc.,
Chem. Commun. 1995, 1809.
9) (a) Perr e´ e-Fauvet, M.; Verchere-B e´ aur, C.; Tarnaud, E.; Anne-
heim-Herbelin, G.; B oˆ ne, N.; Gaudemer, A. Tetrahedron 1996, 52,
3569 and reference therein. (b) J ain, R. K.; Sarracino, D. A.; Richert,
C. Chem Commun. 1998, 423.
(
1

Glycosylated Amino Acid Porphyrin Derivatives
J . Org. Chem., Vol. 64, No. 12, 1999 4433
Sch em e 2a
a
Reaction conditions: (i) BF3OEt2/CH2Cl2, 18 h, then p-choranil; (ii) H2, 10% Pd-C, THF, rt, 5 h; (iii) CH2Cl2, DCC, Fmoc-L-Ala, 15 h,
rt; (iv) morpholine, then NaOMe/MeOH/CH2Cl2.
procedure (Schemes 2 and 3) consists of the condensation
of a suitable aldehyde with pyrrole in the presence of BF
with H
2
/Pd-C in THF for 5 h.13 Glycosylated aminopor-
phyrins 9a ,b, 10, 11, and 12a ,b were obtained after
purification on PLC, in 56-76% yield (Scheme 2). Unlike
glycosylated porphyrins, mononitrophenyltristolylpor-
3
/
etherate (formation of porphyrinogen) followed by oxida-
tion by p-chloranil to afford the porphyrin 3-8. The
condensation of pyrrole (4 equiv) with para nitrobenz-
aldehyde (3 equiv) and 2- or 4-(2′,3′,4′,6′-tetra-O-acetyl-
â-D-glucopyranosyloxy)benzaldehyde 1a ,b (1 equiv) gave
compounds 3a ,b (14% and 16% yield, respectively). The
relative proportion (Table 1) of reagents (pyrrole/4-
nitrobenzaldehyde/1b) have been studied in order to
increase the yields of compounds 3b, 4, 5, and 6b. In all
phyrins were reduced in CHCl
3
/acetic acid by the usual
procedure (SnCl
2
/HCl) (Scheme 3).14 The mixture was
stirred under reflux overnight and then was neutralized
with NaOH. After extraction and purification on PLC,
compounds 13a ,b15 were obtained in 90% yield.
Two series of amino acid porphyrins have been syn-
thesized. The first involves the coupling of 9a ,b, 10, 11,
or 12a ,b with N-Fmoc-L-alanine to give mono-, di-, or tri-
cases, these reagents were added to CH
under argon at room temperature. After usual workup,
2 2
Cl as solvent
amino acid porphyrins glycosylated with protected NH
2
1
d
groups 14a ,b, 15, 16, and 17a ,b (Scheme 2). The second
results from the coupling of 13a ,b with 2 to give mono-
glucosylamino acid porphyrin derivatives 18a ,b with a
porphyrins were purified by a silica gel column and PLC
1
and then individually characterized by H NMR analysis.
For meso-tetraglucosylarylporphyrin 7,1d it was iolated
as a byproduct from the synthesis of 6b. The yields are
shown in Table 1. The synthesis of mononitrophenyltris-
tolylporphyrins 8a ,b are shown in Scheme 3. For the
preparation of these compounds, pyrrole was condensed
with p-tolualdehyde and 2- or 4-nitrobenzaldehyde in a
relative proportion of 4/3/1, but only mononitrophenyl-
tristolylporphyrins were isolated by silica gel column and
purified by PLC. Porphyrins 8a ,b were obtained in 14%
and 15% yield, respectively. The usual reduction of the
protected NH group (Fmoc) (Scheme 3).
The general coupling method involves the formation
of a symmetrical anhydride as a coupling agent with
dicyclohexylcarbodiimide (DCC) and N-Fmoc-L-alanine in
2
2 2
CH Cl (15 h in darkness), which reacts with porphyrin
derivatives. After removing the insoluble dicyclohexyl-
urea by filtration, compounds 14-18 were obtained after
purification on TLC in 90% yield.15 The N-9-fluorenyl-
(
13) Baldwin, Crossley, M. J .; Debernadis, J . Tetrahedron 1982, 38,
85.
(14) Collman, Gagne, R. R.; Reed, C. A.; Halbert, T. R.; Lang, G.;
Robinson, W. T. J . Am. Chem. Soc. 1975, 97, 1427.
15) Tamiaki, H.; Numara, K.; Maryama, K. Bull. Chem. Soc. J pn.
1993, 66, 3062.
6
2
nitro function with SnCl /HCl could not be used because
sugar moieties are sensitive under strongly acidic condi-
tions. So the amino functions were obtained by reduction
(

4
434 J . Org. Chem., Vol. 64, No. 12, 1999
Sol et al.
Sch em e 3a
a
Reaction conditions: (i) BF3OEt2/CH2Cl2, 18 h, then p-chloranil, reflux, 1 h; (ii) SnCl2/HCl, reflux, 15 h; (iii) DCC, 2, 15 h, rt; (iv)
morpholine, then H2N-NH2/MeOH/CH2Cl2.
Ta ble 1. Effect of Ra tio of Rea gen ts on th e Yield s of
pa r a P olyglu cosyla ted 4-Nitr op h en ylp or p h yr in s
Ma ss Ch a r a cter iza tion . Mass spectrometry of all
porphyrin derivatives was performed using the MALDI
4/3/1a
4/2/2a
4/1/3a
19
porphyrins obtained
(matrix-assisted laser desorption ionization) technique.
16b
1.8
1
1
4
Positive ion mass spectra exhibited a base peak corres-
ponding to the intact porphyrin, and no fragment ions
were detected. The analysis of the isotopic components
3
4
5
6
7
b
2
14
10
2
3.5
31.5
1
1
1
16
11
30
b
+
indicated the presence of a protonated species (M + H)
+•
with a minor contribution of the radical cation M
total yield
23.8
(Figure 1), allowing the determination of the molecular
a
Ratio of pyrrole/p-nitrobenzaldehyde/1b. b Yield of pure iso-
mass with an accuracy generally around 0.001%.
lated compounds in %.
1
H NMR Ch a r a cter iza tion . 1H NMR spectra re-
methoxycarbonyl (Fmoc) protective group of the R-amino
function is frequently used in peptide chemistry and was
selectively removed in high yield with a weak base (50%
corded at 400 MHz were used for characterization of
compounds 3-18 in CDCl . The detailed resonance
3
assignments are based on integration and selective
morpholine in CH
literature,17 the acetate groups of glucose moieties (14-
7) were easily removed by treatment at room temper-
ature (1 h) with NaOMe in MeOH/CH Cl (8/2), yielding
2
Cl
2
, rt, 1 h).16 According to the
homonuclear decoupling, as well as NOE and 2D homo-
nuclear COSY experiments. The spectra of these com-
pounds are governed by the symmetry properties of the
product1 and by the orientation of the carbohydrates
3a ,b, 9a ,b, 14a ,b, N-Fmoc-L-alanine 17a ,b, and glyco-
sylated serine 18a ,b. For meso-tetraglucosylarylporphy-
rins 7,1d the resonances of the eight equivalent pyrrolic
protons appear as a single peak at 8.90 ppm. In contrast,
the pyrrolic proton resonances of the others compounds
are more complicated. They depend on the number, the
linking position, and the nature of meso substituents. For
porphyrin derivatives bearing four different mesophenyl
1
e
2
2
compounds 19-22 in 75-64% (Scheme 2). However, for
compounds 18a ,b, deacetylation of the glucose moieties
was not obvious because the usual basic condition leads
to â-elimination.18 We therefore utilized a procedure
developed by Kunz1
8a,b
that relies upon hydrazine in
2 2
MeOH/CH Cl to remove the acetates nucleophilically.
Deacetylation was complete after 2 h at 25 °C, and
compounds 23a ,b were obtained in 71% and 57% yield,
respectively (Scheme 3).
groups with para substitution (3, 6, 7, 9, 12-14, 17, 18)
and according to the literature,1
e,20
the observation and
(16) Carpino, L. A.; Han, G. Y. J . Org. Chem. 1972, 37, 3404.
(17) Zemplen, G.; Gerecs, A.; Hadacsy, I. Berichte 1996, 69, 1827.
(18) (a) Schultheiss-Reimann, P.; Kunz, H. Angew. Chem., Int. Ed.
the comparison of the â pyrrolic protons permits distinc-
tion between three classes of compounds. Indeed, meso
para tetraphenylporphyrin derivatives 3, 6, 8b, 9, 12,
Eng. 1993, 22, 62. (b) Kunz, H. Angew. Chem., Int. Ed. Engl. 1991,
6, 294. (c) Bardaji, E.; Torres, J . L.; Clapes, P.; Albericio, F.; Barany,
2
G.; Valencia, G. Angew. Chem., Int. Ed. Engl. 1991, 29, 291. (d) Polt,
R.; Szubo, L.; Treiberg, J .; Li, Y.; Hruby, J . J . Am. Chem. Soc. 1992,
(19) Karas, M.; Hillenkamp, F. Anal. Chem. 1988, 60, 2299.
(20) Meng, G. G.; J ames, B. R.; Skov, K. A. Can. J . Chem. 1994, 72,
1894.
1
14, 10249.

Glycosylated Amino Acid Porphyrin Derivatives
J . Org. Chem., Vol. 64, No. 12, 1999 4435
F igu r e 1. Example of MALDI mass spectra for compound 22b.
1
3b, 14, 17, and 18b whose meso positions were differ-
and then as two singlets at δ 8.82 and 8.78 ppm for H-12,
-13, -17, and -18. The signals of the nitrophenyl protons
are broadened (Figure 2). These phenomena reflect a
reduced symmetry induced by the orientation of the
substituent that in turn leads to an unsymmetrical folded
structure because of steric hindrance.
ently substituted (three identical meso phenyl groups)
possess a symmetry plane. Thus, the pyrrolic proton
resonance is split into two doublets (J ) 4.9 Hz) and one
singlet because protons H-12,18 and H-13,17, possessing
a quasiequivalent electronic environment, are acciden-
tally equivalent.1d When three meso positions are sub-
stituted by 4-nitrophenyl groups, which are electron-
withdrawing, the resonance structures predict lower
electron densities at H-3, H-7.20 Thus, the doublet at δ
13C NMR An a lysis. 13C NMR spectroscopy was also
used to elucidate the structure by means of DEPT
1
13
experiments and H- C shift correlation. The observa-
tion and the comparison of the â pyrrolic carbons, in meso
para phenyl-substituted compounds (3b, 4, 5, 6b, 7),
permits the distinction of four differents signals. The
appearance and the broadness of these signals are
governed by the symmetry properties of the product,
allowing us to differentiate molecules from each other.
Figure 3 shows the spectra of mono-, bis-, or trisgluco-
sylaryl tri-, di-, or mononitrophenylporphyrins 3b, 4, 5,
and 6b and tetraglucosylarylporphyrin 7 for the â
pyrrolic carbon. In the series of related compounds
characterized by ortho glucosyl, alanyl, and glucosylseryl
substitution (3a , 9a , 14a , 18a ), we found the same
8
.95 is assigned to these two pyrrolic protons, the doublet
at δ 8.78 is assigned to H-2, H-8, and the H-12, H-13,
H-17, H-18 resonance is assigned to the singlet at δ 8.80.
When the three mesophenyl groups are electron-donating
(
as the 4-aminophenyl group 9b, the 4-(N-Fmoc-L-alanyl)-
aminophenyl group 14b, and the 4-glucosylphenyl groups
b, 12b, 17b), they induce an upfield shift for protons
H-2, H-8 (doublet) and for protons H-12, H-13, H-17, H-18
singlet). We have observed the same behavior of â
6
(
pyrrolic protons for tristolylporphyrin derivatives whose
chemical shifts depend on the electronic effect of the
4
1
-nitrophenyl 8b, 4-aminophenyl 13b, or 4-glucosylseryl-
phenomenon as in H NMR, that is, a shielding of ortho
aminophenyl 18b groups. The two other classes of
porphyrins were substituted by two identical meso phenyl
groups. The observation of their pyrrole protons permits
us to distinguish between cis and trans isomers readily
substituents and a modification of â pyrrolic carbons that
reflect a diminished symmetry of the compounds and a
distortion of the porphyrin macrocycle.
UV-visible Absor p tion . The porphyrins synthesized
in this work show typical electronic spectra, with a Soret
1
by H NMR. Thus, pyrrole protons of trans compounds
(4, 10, 15) (Scheme 2) which are the most symmetrical,
band near 420 nm and four less intense visible bands Q
give two doublets for any pair of adjacent protons. For
the cis isomer (5, 11, 16) (Scheme 2), the four types of
pyrroles protons give rise to two singlets (H-7(8), H-17-
21
(
I, II, III, and IV) in CH
2
Cl
2
.
Generally, these porphyrins
), but there are a few
exceptions. The relative intensities of the Q (II) and Q
I) band of certain porphyrin derivatives 9a ,b, 10, 11,
2b, 14b, 15 show a nonstandard type (ꢀ
give etio spectra (ꢀIV > ꢀIII > ꢀII > ꢀ
I
(18)) and two coupled doublets (H-2(13), H-3(12)).
(
1
For porphyrin derivatives which possess only one meso
I
/ꢀII > 1). In
phenyl group with ortho substitution (3a , 6a , 8a , 9a , 12a ,
3a , 14a , 17a , 18a ), we have observed an obvious change
aqueous solutions, glucosylseryltristolylporphyrins 23a ,b
1
exhibited an extremely broadened split Soret band
1
f
in the chemical shift of the phenyl ring. Furthermore,
compounds (3a , 9a , 14a , 18a ) which are characterized
by ortho glucosyl, alanyl, and glucosylseryl substitutions,
(Figure 4). This result is due to the combination of
cofacial and edge-to-edge interaction of self-assembled
22
aggregates. In contrast, the Soret band of compound
1
are subject to considerable changes in the H NMR
2
2b is broadened and blue-shifted, whereas the Soret
chemical shifts and/or in figures relative to most of the
nuclei. Thus, for example, all the sugar protons experi-
ence a particularly pronounced shielding from -0.31 ppm
band of alanylphenylglucosylarylporphyrins 19-22a are
split into two bands that are also blue-shifted (Table 2).
These blue-shift absorptions could be attributed to face-
(H-6
A
) to -1.31 ppm (H-2), and two acetyl groups that
23
to-face orientation.
would otherwise show a larger shift toward 2.0 ppm are
also shifted downfield (δ 1.26 and δ -0.92). These nuclei
are obviously located well within the range of the
shielding current above the porphyrin macrocycle. More-
over, the â pyrrole proton signals of 3a appeared as four
doublets (J ) 4.9 Hz) at 8.74, 8.77, 8.81, and 8.92 ppm
(
21) Falk, J . E. Porphyrins and metalloporphyrins; Elsevier: Am-
sterdam, 1964.
22) Furhop, J . H.; Demoulin, C.; Boettcher, C.; Koning, J .; Siggel,
U. J . Am. Chem. Soc. 1992, 114, 4159.
23) Barber, D. C.; Freitag-Beeston, R. A.; Whiten, D. C. J . Phys.
Chem. 1991, 95, 4074.
(
(

4
436 J . Org. Chem., Vol. 64, No. 12, 1999
Sol et al.
F igu r e 2. Behavior of â pyrrolic (δ 8.7-9.0 ppm) and meso glucosylaryl (7.4-8.7 ppm) protons for 3a ,b compounds.
F igu r e 3. â pyrrole carbon part of 13C NMR spectra of compounds 3-7. The structures indicate the molecular symmetry.
F lu or escen ce Sp ectr oscop y. All fluorescence spectra
of compounds in CH Cl were characterized by two
emission bands.1 The only porphyrins (6a , 8a ) bearing
an NO group in the ortho position gave no fluorescence
emission, most likely as a result of specific interaction
of the NO function with the porphyrin ring, whereas
isomers 6b and 8b with the NO group in the para
position showed a significant fluorescence emission. The
fluorescence emission wavelengths of 20, 21, and 22a ,b
in aqueous solutions were identical to those obtained in
THF, but the emission was strongly quenched. This decay
of fluorescence can be explained by the formation of
aggregates and supports the results observed in the
UV-visible spectra.
Sin glet Oxygen P r od u ction . To determine the pho-
2
2
i
22
2
2
tosensitizing properties of porphyrins 19-23, the trap-
1
2
ping reactions of O
2
with ergosterol acetate were carried
out.24 Reference experiments with eosin and HP as
(24) Ohloff, G. Pure Appl. Chem. 1975, 43, 481.

Glycosylated Amino Acid Porphyrin Derivatives
J . Org. Chem., Vol. 64, No. 12, 1999 4437
Ta ble 2. UV-vis Sp ectr a of P or p h yr in Der iva tives in Va r iou s Solven ts
λnm (ꢀ × 10 3, cm-1 mol-1 L)
-
compd
solvent )
a
(
Soret 1
Soret 2
IV
III
II
I
ratio ꢀI/ꢀII
3
3
4
5
6
6
7
8
8
9
9
a (a)
b (a)
(a)
422 (380.3)
424 (152.0)
422 (279.5)
422 (380.3)
420 (233.3)
420 (335.9)
420 (458.0)
420 (365.4)
420 (325.4)
426 (205.9)
426 (434.7)
424 (384.2)
424 (376.5)
422 (344.8)
422 (312.0)
420 (435.7)
422 (335.7)
422 (309.1)
422 (240.9)
422 (319.6)
422 (352.7)
422 (240.6)
420 (348.6)
420 (473.4)
420 (379.0)
420 (46.3)
518 (10.6)
516 (12.7)
516 (16.6)
518 (10.6)
516 (13.3)
516 (13.9)
516 (17.6)
516 (16.7)
518 (12.3)
520 (10.9)
522 (11.0)
520 (16.3)
520 (14.9)
518 (11.7)
518 (13.4)
516 (19.3)
518 (16.5)
516 (11.5)
518 (11.4)
518 (8.8)
554 (5.9)
552 (6.8)
554 (9.5)
554 (5.9)
552 (8.2)
552 (6.8)
552 (9.8)
552 (7.8)
554 (9.2)
560 (9.0)
562 (10.4)
558 (13.3)
558 (12.2)
554 (7.5)
554 (9.1)
552 (10.1)
554 (5.4)
552 (6.1)
554 (7.7)
554 (5.5)
554 (7.0)
552 (6.3)
552 (6.8)
552 (9.0)
552 (5.3)
558 (6.4)
558 (6.5)
560 (6.0)
558 (6.8)
560 (6.5)
560 (6.3)
554 (6.3)
556 (5.6)
592 (3.5)
590 (4.2)
592 (5.3)
592 (3.5)
592 (5.0)
592 (4.3)
592 (5.5)
592 (5.0)
592 (4.3)
596 (4.0)
596 (4.0)
594 (5.5)
594 (5.2)
594 (4.9)
592 (4.3)
592 (6.0)
594 (2.6)
592 (3.5)
592 (3.9)
594 (2.7)
592 (4.9)
592 (3.7)
592 (3.6)
590 (5.3)
592 (2.2)
598 (3.3)
600 (3.6)
598 (3.1)
600 (3.8)
600 (3.7)
598 (3.7)
594 (3.8)
596 (3.2)
648 (2.5)
646 (2.5)
648 (4.2)
648 (2.5)
648 (4.2)
648 (2.9)
648 (4.4)
648 (3.8)
648 (3.0)
654 (5.1)
654 (5.5)
652 (7.1)
652 (6.5)
650 (4.5)
650 (4.5)
648 (4.0)
650 (2.1)
648 (3.2)
648 (3.8)
650 (2.8)
648 (4.8)
650 (3.6)
648 (3.5)
646 (4.1)
648 (2.0)
654 (3.3)
654 (3.5)
652 (3.0)
654 (3.6)
654 (3.6)
652 (3.5)
652 (3.5)
652 (3.1)
0.71
0.59
0.79
0.71
0.84
0.67
0.80
0.76
0.70
1.27
1.37
1.29
1.25
0.92
1.05
0.67
0.81
0.91
1.02
1.04
0.98
0.97
0.97
0.77
0.91
1.00
0.97
0.97
0.95
0.97
0.95
0.92
0.97
(a)
a (a)
b (a)
(a)
a (a)
b (a)
a (a)
b (a)
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
2
2
2
2
2
2
0 (a)
1 (a)
2a (a)
2b (a)
3a (a)
3b (a)
4a (a)
4b (a)
5 (a)
6 (a)
518 (9.5)
7a (a)
7b (a)
8a (a)
8b (a)
9a (b)
9b (b)
0 (b)
516 (10.9)
516 (11.8)
516 (17.2)
516 (9.3)
406 (75.6)
404 (79.4)
404 (73.1)
406 (82.1)
396 (70.4)
522 (7.9)
b
420 (65.4)
522 (8.1)
422 (44.1)
522 (7.1)
b
1 (b)
422 (60.1)
522 (8.9)
2a (b)
2b (b)
3a (b)
3b (b)
416 (40.1)
416 (79.6)
450 (50.3)
450 (42.9)
526 (8.1)
524 (8.0)
b
414 (17.1)
526 (12.4)
526 (10.1)
b
416 (16.6)
a
The solvents used are as follows: (a) CH2Cl2, (b) aqueous solutions. b Soret band appears as a “shoulder”.
lower than with HP, and did not increase with illumina-
tion times. Moreover, these dead cell counts were identi-
cal even if the irradiation time increased. Overall, the
2
4 h incubation in the dark resulted in an increase of
dead cell counts. Although the immediate effects account
for early necrotic death, the delayed postirradiation
deaths are likely the result of photoirradiation-induced
apoptosis followed by secondary necrosis. Although com-
pounds 20, 21, and 22a containing alanyl groups induced
limited immediate cell death when compared to the
immediate effect of HP, they lead to a large percentage
of dead cells after 24 h postirradiation incubation in the
dark, provided that the incubation is longer than 60 min.
All other synthetic compounds showed a lower effect,
especially compounds 23a ,b containing three tolyl groups
and one glucosylseryl moiety, for which postirradiation
dead cell count stalled as soon as they were irradiated
for 40 min. Differences in activities of these glucosylated
amino acid porphyrin derivatives may be attributed to
the nature, number, and meso positions of amino acids
and glucose.
-
5
F igu r e 4. Absorption spectra of porphyrins 23b (3.3 × 10
M) in THF (- - -) and in aqueous solution (s).
sensitizers gave ergosterol acetate endoperoxide with
nearly quantitative yields. In the same experimental
conditions, porphyrins 19-23 were almost as efficient as
HP, which is known as a photosensitizer that produces
singlet oxygen.
Biologica l E ssa ys. The photocytotoxicity of gluco-
sylated amino acid porphyrin derivatives 19a ,b and
2a ,b have been described in a previous paper. Using
the same conditions, the photocytotoxicity of compounds
0, 21, and 23a ,b was tested against K562 human
chronic myelogenous leukemia cells. Figure 5 displays
dead cell counts as a function of irradiation time and the
subsequent increase following a further 24 h incubation
in the dark. HP was used as a control. For all synthetic
porphyrins, the immediate dead cell counts were always
In summary, this paper describes the synthesis and
the characterization of glucosylaminoacylporphyrin deriv-
atives that are obtained in best yield by condensation in
different proportions of two benzaldehydes with pyrrole
using Lindsey’s conditions. To improve cell membrane
recognitions, these tetrapyrrolic macrocycles are substi-
tuted on the meso positions by amino acids and glucose.
Substitutions in the ortho and para positions lead to
further insights into cell membrane interaction. The
photocytotoxicity of these synthetic porphyrins against
K562 indicated that the immediate dead cell counts were
always lower than those observed with HP. However, for
three porphyrin derivatives, the delayed effects (24 h
1
i
2
2

4
438 J . Org. Chem., Vol. 64, No. 12, 1999
Sol et al.
F igu r e 5. Percentage of PI stained cell vs time. Open bars: dead cell count after indicated irradiation time. Solid bars: dead cell
count after a further 24 h incubation in the dark.
incubation in the dark at 37 °C) were always comparable
to those observed with HP when irradiation was longer
than 60 min. The present strategy may be easily used
for preparation of glycosylated porphyrins substituted on
the meso positions by small peptides (three or four amino
acids).
temperature. p-Chloranil (0.75 equiv/pyrrole) was then added,
and the mixture was stirred under reflux for 1 h. The solvent
was evaporated to dryness, and the porphyrin mixture was
purified by column chromatography (toluene/acetone, 100/0 to
7
0/30) and thin-layer chromatography.
-[2-(2′,3′,4′,6′-Tetr a -O-a cetyl-â-D-glu cop yr a n osyloxy)-
p h en yl]-10,15,20-tr is(4-n itr op h en yl) P or p h yr in (3a ). 1a
452 mg, 1 mmol, 1 equiv), 4-nitrobenzaldehyde (453 mg, 3
mmol, 3 equiv), and pyrrole (0.28 mL, 4 mmol, 4 equiv)
afforded pure product 3a 154 mg (14%). R 0.41 (toluene/
acetone, 85/15). UV-visible (see Table 2). H NMR (CDCl
00 MHz) δ -2.80(2H, br s, NH); -0.92(3H, s, OAc); 1.26(3H,
s, OAc); 1.86(3H, s, OAc); 1.96(3H, s, OAc); 3.65(1H, ddd, J )
5
(
Exp er im en ta l Section
f
Gen er a l. All solvents and reagents were purchased from
1
3
,
Aldrich, Prolabo, or J anssen. Pyrrole was distilled over CaH
under reduced pressure immediately before use. Methylene
chloride and chloroform were distilled over P and then
CaH . Analytical thin-layer chromatography (TLC) was per-
formed on silica gel (Merck, 60F254). Merck precoated plates
silica gel 60, 2 mm) were used for preparative thin-layer
2
2
2
O
5
7
4
2
7
.7, 5.1, 2.5 Hz, H-5′ose); 4.00(1H, dd, J ) 12.1-2.6, H-6
b
′ose);
2
.12(1H, dd, J ) 12.1-5.2 Hz, H-6 ′ose); 4.18(1H, dd, J ) 7.7-
a
.2 Hz, H-2′); 4.64(1H, t, J ) 9.2 Hz, H-3′ose); 4.92(1H, d, J )
.7 Hz, H-1′ose); 7.52(1H, dt, J ) 7.5-1.6 Hz, H-5 aryl); 7.70-
(
chromatography. Column chromatography was carried out
with silica gel (60 ACC, 15-40 µm, Merck) or with Sephadex
LH20 (Pharmacia). The UV-visible spectra were obtained on
a spectrophotometer using 1 or 0.1 cm quartz cells, and
emission spectra were recorded by using a Perkin-Elmer LS-
(
1H, dt, J ) 7.5-1.6 Hz, H-3 aryl); 7.80(1H, dt, J ) 7.5-1.6
Hz, H-4 aryl); 8.05(1H, dd, J ) 7.5-1.6 Hz, H-6 aryl); 8.40-
6H, m, H-2,6 nitrophenyl); 8.65(6H, br d, J ) 8.6 Hz, H-3,5
nitrophenyl); 8.74(1H, d, J ) 4.9 Hz, H-2 or H-8 â pyrrole);
.77(1H, d, J ) 4.9 Hz, H-8 or H-2 â pyrrole); 8.78(2H, s, H-12,-
(
8
5
B spectrofluorimeter. Melting points were determined by
1
13
18 or H13,17 â pyrrole); 8.81(1H, d, J ) 4.9 Hz, H-3 or H-7 â
pyrrole); 8.82(2H, s, H-12,18 or H-13,17 â pyrrole); 8.92(1H,
capillary tube apparatus and were not corrected. H and
C
NMR spectra were recorded in CDCl with tetramethylsilane
3
1
3
d, J ) 4.9 Hz, H-7 or H-3 â pyrrole). C NMR (CDCl
7.5(1C, CH CO); 19.8(1C, CH CO); 20.3(1C, CH
1C, CH CO); 62.1(1C, C-6′ose); 68.4(1C, C-4′ose); 70.1(1C,
C-2′ose); 72.0(1C, C-5′ose); 72.3(1C, C-3′ose); 99.7(1C, C-1′ose);
16.6(1C, C meso); 117.6(1C, C meso); 117.7(1C, C meso); 117.8-
1C, C-3 aryl); 118(1C, C meso); 121.9(6C, C-3,5 nitrophenyl);
22.7(1C, C-5 aryl); 130.4(C â pyrrole and C-4 aryl); 131.7(C
3
, 50 MHz)
CO); 20.5-
as an internal standard. The chemical shifts are given in ppm,
and coupling constants are in Hz. MALDI mass spectra were
obtained on a Voyager Elite (Framingham, MA) time-of-flight
mass spectrometer. E.I. mass spectra were performed at the
1
(
3
3
3
3
1
(
1
“Laboratoire D e´ partemental d’Analyse de Limoges” and el-
emental analyses were carried by the “Service R e´ gional de
Microanalyse de l’Universit e´ Pierre et Marie Curie, Paris”.
Syn th esis. Benzaldehydes 1a ,b and glycosylated serine
â pyrrole); 132.2(1C, C-1 aryl); 135.0(6C, C-2,6 nitrophenyl);
135.9(1C, C-6 aryl); 146.2(8C, C-R pyrrole); 147.9(3C, C-4
nitrophenyl); 148.5(3C, C-1 nitrophenyl); 156.2(1C, C-2 aryl);
derivative 2 were synthesized according to the literature.1
f,10,11
Syn th esis of Nitr op h en ylp or p h yr in s. Pyrrole (4 equiv),
per-O-acetyl glucosylated benzaldehyde 1a ,b (1, 2, or 3 equiv)
or p-tolualdehyde (3 equiv), and 2- or 4-nitrobenzaldehyde (1,
1
1
66.9(1C, CH
3
CO); 169.0(1C, CH
CO). MS (MALDI) m/z: 1096.2 ([M + H] mono-
16: C, 63.55; H, 4.14; N,
.94. Found: C, 63.35; H, 4.16; N, 8.92.
3 3
CO); 169.5(1C, CH CO);
+
70.3(1C, CH
3
isotopic). Anal. Calcd for C58
8
45 7
H N O
2
, or 3 equiv) were added to methylene chloride (500 mL)
purged with argon for 30 min. The mixture was stirred and
purged with argon for a further 10 min, after which a BF
3
/
5-[4-(2′,3′,4′,6′-Tetr a -O-a cetyl-â-D-glu cop yr a n osyloxy)-
p h en yl]-10,15,20-tr is(4-n itr op h en yl) P or p h yr in (3b). 1b
(452 mg, 1 mmol, 1 equiv), 4-nitrobenzaldehyde (453 mg, 3
etherate solution (0.5 mL, 0.2 M) in methylene chloride was
added. This reaction mixture was stirred overnight at room

Glycosylated Amino Acid Porphyrin Derivatives
J . Org. Chem., Vol. 64, No. 12, 1999 4439
mmol, 3 equiv), and pyrrole (0.28 mL, 4 mmol, 4 equiv) gave
equiv), and pyrrole (0.28 mL, 4 mmol, 4 equiv) gave 6a 238
3
(
b 175 mg (16%); R
f
0.53 (toluene/acetone, 85/15). UV-visible
see Table 2). H NMR (CDCl , 200 MHz) δ -2.77(2H, br s,
NH); 2.12(3H, s, OAc); 2.13(3H, s, OAc); 2.14(3H, s, OAc); 2.23-
mg (14%). R
f
0.45 (toluene/acetone, 60/40). UV-visible (see
1
1
3
Table 2). H NMR (CDCl , 200 MHz) δ -2.71(2H, br s, NH);
3
2.11(9H, s, -OAc); 2.12(9H, s, -OAc); 2.13(6H, s, OAc); 2.14-
(3H, s, -OAc); 2.23(9H, s, -OAc); 4.08(3H, ddd, J ) 9.7-5.3-
(
(
5
7
3H, s, OAc); 4.08(1H, ddd, J ) 9.6-5.2-2.6 Hz, H-5′ ose); 4.32-
1H, dd, J ) 12.3-2.6 Hz, H-6 ′ose); 4.43(1H, dd, J ) 12.3-
′ose); 5.35(1H, m, H-4′ose); 5.49(3H, m, H-1′,2′,3′ose);
.43(2H, d, J ) 8.6 Hz, H-3,5 aryl); 8.15(2H, d, J ) 8.6 Hz,
b
2.8 Hz, H-5′ ose); 4.31(3H, dd, J ) 12.2-2.2, H-6
4.43(3H, dd, J ) 12.2-5.3 Hz, H-6
b
′ ose);
′ ose); 5.34(3H, br d, J )
.2 Hz, H-6
a
a
9.7 Hz, H-4′ ose); 5.48(9H, m, H-1′,2′,3′ ose); 7.40(6H, br d, J
) 7.3 Hz, H-3,5 aryl); 7.96 (2H, m, H-4,5 o-nitrophenyl); 8.14
(6H, brd, J ) 7.5 Hz, H-2,6 aryl); 8.25(1H, dd, J ) 7.1-2.1
Hz, H-6 o-nitrophenyl); 8.44(1H, dd, J ) 7.3-2.1 Hz, H-3
o-nitrophenyl); 8.67(2H, d, J ) 4.9 Hz, H-3,7 â pyrrole); 8.85-
(2H, d, J ) 4.8 Hz, H-2,8 â pyrrole); 8.86(4H, s, H-12,13,17,18
H-2,6 aryl); 8.38(6H, d, J ) 8.9 Hz, H-2,6 nitrophenyl); 8.65-
6H, d, J ) 8.6 Hz, H-3,5 nitrophenyl); 8.78(2H, d, J ) 4.9 Hz,
H-2,8 â pyrrole); 8.80(4H, s, H-12,13,17,18 â pyrrole); 8.95-
(
1
3
(
2
2H, d, J ) 4.9 Hz, H-3,7 â pyrrole). C NMR (CDCl
3
, 50 MHz)
0.6(2C, CH CO); 20.8(2C, CH CO); 62.1(1C, C-6′ose); 68.4-
3
3
(
(
1C, C-4′ose); 71.4(1C, C-2′ or 3′ose); 72.3(1C, C-5′ose); 72.8-
1C, C-2′ or 3′ ose); 99.1(1C, C1′ose); 115.6(2C, C-3,5 aryl);
â pyrrole). 13C NMR (CDCl
(6C, CH
3
, 50 MHz) 20.6(6C, CH
3
CO); 20.7-
3
CO); 62.1(3C, C-6′ose); 68.4(3C, C-4′ose); 71.3(3C, C-2′
1
(
1
17.7(1C, C meso); 118.0(2C, C meso); 121.1(2C, C meso); 122.0-
6C, C-3,5 nitrophenyl); 131.2(C â pyrrole); 132.3(C â pyrrole);
35.0(6C, C-2,6 nitrophenyl); 135.6(2C, C-2,6 aryl); 136.4(1C,
C-1 aryl); 146.2(8C, C R pyrrole); 147.6(3C, C-4 nitrophenyl);
48.5(3C, C-1 nitrophenyl); 156.9(1C, C-4 aryl); 168.3(1C,
or C-3′ose); 72.3(3C, C-5′ose); 72.6(3C, C-2′ or C-3′ose); 99.1-
(3C, C-1′ose); 113.9(1C, C meso); 115.1(6C, C-3,5 aryl); 119.6-
(2C, C meso); 120,0(1C, C meso); 124,0(1C, C-3 nitrophenyl);
129.5(1C, C-4 nitrophenyl); 130.9(1C, C-5 nitrophenyl); 131.1-
(C-â pyrrole); 131.5(C-â pyrrole); 135.5(6C, C-2,6 aryl); 136.5-
(1C, C-6 nitrophenyl); 136.8(1C, C-1 nitrophenyl); 136.9(2C,
C-1 aryl); 137.1(1C, C-1 aryl); 146.5(8C, C-R pyrrole); 169.4
1
CH
3
CO); 169.4(1C, CH
CO). MS (MALDI) m/z: 1096.2 ([M + H] monoisotopic).
16: C, 63.55; H, 4.14; N, 8.94. Found:
C, 63.48; H, 4.11; N, 8.91.
,10,15,20-B is (4-n it r o p h e n y l)-b is [(2′,3′,4′,6′-t e t r a -
3 3
CO); 170.3(1C, CH CO); 170.5(1C,
+
CH
3
Anal. Calcd for C58
H
45
N
7
O
(6C, CH
(MALDI) m/z: 1699.5 ([M + H] monoisotopic). Anal. Calcd for
O: C, 59.49; H, 5.13; N, 4.00. Found: C, 59.55;
H, 5.15; N, 4.02.
3 3 3
CO); 170.2(4C, CH CO); 170.5(2C, CH CO). MS
+
5
C
86
H
83
N
5
O
32‚2H
2
O-a cetyl-â-D-glu cop yr a n osyloxy)p h en yl] P or p h yr in (4)
a n d (5). 1b (904 mg, 2 mmol, 2 equiv), 4-nitrobenzaldehyde
5-(4-Nitr oph en yl)-10,15,20-tr is[4-(2′,3′,4′,6′-tetr a-O-acetyl-
â-D-glu cop yr a n osyloxy)p h en yl] P or p h yr in (6b). 1b (1.35
g, 3 mmol, 3 equiv), 4-nitrobenzaldehyde (151 mg, 1 mmol, 1
equiv), and pyrrole (0.28 mL, 4 mmol, 4 equiv) gave 6b 272
(
302 mg, 2 mmol, 2 equiv), and pyrrole (0.28 mL, 4 mmol, 4
equiv) gave 4 196 mg (14%) and 5 140 mg (10%). 4: R 0.42
toluene/acetone, 75/25, two elutions). UV-visible (see Table
f
(
2
1
). H NMR (CDCl
3
, 200MH) δ -2.80(2H, br s, NH); 2.04(6H,
mg (16%). R
f
0.50 (toluene/acetone, 85/15). UV-visible (see
1
s, OAc); 2.05(12H, s, OAc); 2.06(6H, s, OAc); 4.08(2H, ddd, J
9.6-5.2-2.5 Hz, H-5′ose); 4.32(2H, J ) 12.3-2.6 Hz,
H-6 ′ose); 5.33(2H, m,
Table 2). H NMR (CDCl , 200 MHz) δ -2.77(2H, br s, NH);
3
)
2.12(9H, s, -OAc); 2.13(18H, s, -OAc); 2.23(9H, s, -OAc);
4.08(3H, ddd, J ) 9.8-5.3-2.6 Hz, H-5′ ose); 4.32(3H, dd, J )
b
′ose); 4.42(2H, J ) 12.3-5.4 Hz, H-6
a
H-4′ose); 5.47(6H, m, H-1′,2′,3′ose); 7.40(4H, d, J ) 8.6 Hz,
H-3,5aryl); 8.13(4H, d, J ) 8.6 Hz, H-2,6 aryl); 8.37(4H, d, J
12.1-2.1 Hz, H-6′
b
ose); 4.42(3H, dd, J ) 12.1-5.1 Hz, H-6′
a
ose); 5.35(3H, m, H-4′ ose); 5.49(9H, m, H-1′,2′,3′ ose); 7.39-
(6H, d, J ) 8.5 Hz, H-3,5 aryl); 8.14(6H, d, J ) 8.4 Hz, H-2,6
aryl); 8.39(2H, d, J ) 8.6 Hz, H-2,6 nitrophenyl); 8.64(2H, d,
J ) 8.6 Hz, H-3,5 nitrophenyl); 8.75(2H, d, J ) 4.9 Hz, H-3,7
â pyrrole); 8.88(4H, s, H-12,13,17,18 â pyrrole); 8.90(2H, d, J
)
8.7 Hz, H-2,6 nitrophenyl); 8.63(4H, d, J ) 8.7 Hz, H-3,5
nitrophenyl); 8.75(4H, d, J ) 4.8 Hz, H-3,7,13,17 â pyrrole);
1
3
8
.91(4H, d, J ) 4.8 Hz, H-2,8,12,18 â pyrrole). C NMR
(CDCl , 50 MHz) 20.5(8C, CH CO); 62.0(2C, C-6′ose); 68.3(2C,
3
3
1
3
C-4′ose); 71.3(2C, C-2′ or C-3′ose); 72.5(2C, C-5′ose); 72.7(2C,
C-2′ or C-3′ose); 99.0(2C, C-1′ose); 115.2(4C, C-3,5 aryl); 117.5-
) 4.9 Hz, H-2,8 â pyrrole). C NMR (CDCl
3
, 50 MHz) 20.6-
(8C, CH CO); 20.8(4C, CH CO); 62.1(3C, C-6′ ose); 68.4(3C,
3
3
(
1
(
2C, C meso); 120.2(2C, C meso); 121.6(4C, C-3,5 nitrophenyl);
31.7(8C, C â pyrrole); 135.0(4C, C-2,6 nitrophenyl); 136.5-
2C, C-1 aryl); 146.2(8C, C R pyrrole); 147.8(2C, C-4 nitrophe-
nyl); 148.7(2C, C-1 nitrophenyl); 161.2(2C, C-4 aryl); 169.3(4C,
CH CO); 170.2(2C, CH CO); 170.5(2C, CH CO). MS (MALDI)
m/z: 1397.4 ([M + H]+ monoisotopic); Anal. Calcd for
O: C, 60.43; H, 4.78; N, 5.87. Found: C, 60.25;
H, 4.80; N, 5.83. 5: 0.44 (toluene/acetone,75/25, two
elutions). UV-visible (see Table 2). H NMR (CDCl
δ -2.77(2H, br s,NH); 2.12(6H, s, OAc); 2.13(6H, s, OAc); 2.14-
6H, s, OAc); 2.23(6H, s, OAc); 4.08(2H, ddd, J ) 9.6-5.2-2.5
Hz, H-5′ose); 4.33(2H, dd, J ) 12.5-2.5, H-6 ′ose); 4.45(2H,
′ose); 5.33(2H, m, H-4′ose); 5.48-
6H, m, H-1′,2′,3′ose); 7.14(4H, d, J ) 8.6 Hz,H-3,5 aryl); 8.14-
C-4′ ose); 71.4(3C, C-2′ or C-3′ ose); 72.3(3C, C-5′ ose); 72.8-
(3C, C-3′ or C-2′ ose); 99.1(3C, C-1′ ose); 115.2(6C, C-3,5 aryl);
116.8(1C, C meso); 119.2(2C, C meso); 120.2(1C, C meso); 121.7-
(2C, C-3,5 nitrophenyl); 130.3(C-â pyrrole); 131.7(C-â pyrrole)
135.1(2C, C-2,6 nitrophenyl); 135.5(6C, C-2,6 aryl); 136.7(3C,
C-1 aryl); 146.2(8C, C-R pyrrole); 147.8(1C, C-4 nitrophenyl);
149.1(1C, C-1 nitrophenyl); 156.7(3C, C-4 aryl); 168.4(6C,
3
3
3
C
72
H
64
N
6
O
24‚2H
2
R
f
CH
m/z: 1699.5 ([M
32: C, 60.81; H, 4.92; N, 4.12. Found: C, 60.67; H,
3
CO); 170.3(3C, CH
3
CO); 170.6(3C, CH
3
CO). MS (MALDI)
1
+
3
, 200 MHz)
+ H] monoisotopic). Anal. Calcd for
86 83 5
C H N O
(
4.89; N, 4.09.
b
5,10,15,20-Tetr a [4-(2′,3′,4′,6′-tetr a -O-a cetyl-â-D-glu cop y-
r a n osyloxy)p h en yl] P or p h yr in (7).1d This compound was
isolated as a byproduct from the synthesis of 6b. 220 mg were
dd, J ) 12.5-5.2 Hz, H-6
(
(
nitrophenyl); 8.64(4H,d,H-3,5 nitrophenyl); 8.76(2H,d, J ) 4.9
Hz,H-3,12 â pyrrole); 8.78(2H,s,H-7,8 â pyrrole); 8.91(2H,s,H-
a
4H,d, J ) 8.6 Hz,H-2,6 aryl); 8.38(4H,d, J ) 8.6 Hz,H-2,6
f
obtained (11%). R 0.49(toluene/acetone/methanol, 80/10/10).
1
UV-visible (see Table 2). H NMR (CDCl , 200 MHz) -2.78-
3
(2H, br s, NH); 2.11(3H, s, OAc); 2.12(3H, s, OAc); 2.13(3H, s,
1
3
1
7,18 â pyrrole); 8.93(2H, d, J ) 4.9 Hz, H-2,13 â pyrrole).
NMR (CDCl , 50 MHz) 20.6(4C, CH CO); 20.8(4C, CH CO);
2.1(2C, C-6′ose); 68.4(2C, C-4′ose); 71.3(2C, C-2′ose or C-3′ose);
C
OAc); 4.08(4H, ddd, J ) 2.4-5.3-9.6 Hz, H-5′ ose); 4.32(4H,
3
3
3
dd, J ) 2.4-5.3-9.6 Hz, H-6 ′ose); 4.44(4H, dd, J ) 5.3-12.3
b
6
7
1
1
Hz, H-6′a ose); 5.35(4H, m, H-4′ ose); 5.49(12H, m, H-1′,2′,3′
ose); 7.40(8H, d, J ) 8.6 Hz, H-3,5 aryl); 8.14(8H, d, J ) 8.5
2.3(2C, C-5′ose); 72.6(2C, C-2′or C-3′ose); 99.1(2C, C-1′ose);
15.2(4C, C-3,5 aryl); 117.2(2C, C meso); 120.7(2C, C meso);
21.9(4C, C-3,5 nitrophenyl); 130.9(C-â pyrrole); 131.6(C-â
Hz, H-2,6 aryl); 8.90(8H, s, H-â pyrrole). 13C NMR (CDCl
, 50
3 3
MHz) 26.6(3C, CH CO); 20.8(1C, CH CO); 62.1(4C, c-6′ ose);
3
pyrrole); 136.4(2C, C-1 aryl); 135.0(4C, C-2,6 nitrophenyl);
35.5(4C, C-2,6 aryl); 146.2(8C, C-R pyrrole); 147.6(2C, C-4
nitrophenyl); 148.7(2C, C-1 nitrophenyl); 156.8(2C, C-4 aryl);
69.4(4C, CH CO); 170.3(2C, CH CO); 170.6(2C, CH CO). MS
MALDI) m/z: 1397.4 ([M + H] monoisotopic). Anal. Calcd for
O: C, 59.67; H, 4.87; N, 5.80. Found: C, 59.87;
H, 4.90; N, 5.84.
-(2-Nitr oph en yl)-10,15,20-tr is[4-(2′,3′,4′,6′-tetr a-O-acetyl-
68.4(4C, C-4′ ose); 71.4(4C, C-2′ ose or C-3′ ose); 72.3(4C, C-5′
ose); 72.8(4C, C-3′ ose or C-2′ ose); 99.2(4C, C-1′ ose); 115.1-
(8C, C-3,5 aryl); 119.3(4C, C meso); 131.1(8C, C-â pyrrole);
135.5(8C, C-2,6 aryl); 137.5(4C, C-1 aryl); 146.2(8C, C-R
1
1
(
C
3
3
3
+
pyrrole); 156.6(4C, C-4 aryl); 169.4(2C, CH
CH CO); 170.5(1C, CH CO). MS (MALDI) m/z: 1999.9 ([M +
H] monoisotopic). Anal. Calcd for C100 40: C, 60.06; H,
3
CO); 170.3(1C,
72
H
64
N
6
O
24‚3H
2
3
3
+
H
102 4
N O
5
5.14; N, 2.80. Found: C, 60.04; H, 5.17; N, 2.82.
â-D-glu cop yr a n osyloxy)p h en yl] P or p h yr in (6a ). 1a (1.35
g, 3 mmol, 3 equiv), 2-nitrobenzaldehyde (151 mg, 1 mmol, 1
5-(2-Nitr oph en yl)-10,15,20-tr istolylpor ph yr in (8a). 2-Ni-
trobenzaldehyde (151 mg, 1 mmol, 1 equiv), p-tolualdehyde

4
440 J . Org. Chem., Vol. 64, No. 12, 1999
Sol et al.
(
0.35 mL, 3 mmol, 3 equiv), and pyrrole (0.28 mL, 4 mmol, 4
equiv) gave 8a 105 mg (15%). R 0.75 (CHCl ). UV-visible (see
, 200 MHz) δ -2.70 (2H, s, NH); 2.71
5-[4-(2′,3′,4′,6′-Tetr a -O-a cetyl-â-D-glu cop yr a n osyloxy)-
p h en yl]-10,15,20-tr is(4-a m in op h en yl) P or p h yr in (9b). 3b
(30 mg, 0.027 mmol) and 10% Pd/C (172 mg) gave 15 mg of 9b
f
3
1
Table 2). H NMR (CDCl
3
(
(
(
2
8
9H, s, Me tolyl); 7.56 (6H, d, J ) 7.9 Hz, H-3-5 tolyl); 7.94
(56%). R
(see Table 2). H NMR (CDCl
NH); 2.10(3H, s, OAc); 2.11(3H, s, OAc); 2.12(3H, br s, OAc);
2.22(3H, br s, OAc); 4.03(2H, br s, NH ); 4.05(1H, ddd, J )
9.6-5.5-2.4 Hz, H-5′ose); 4.30(1H, dd, J ) 12.3-2.3 Hz,
H-6 ′ose); 4.41(1H, dd, J ) 12.2-5.5 Hz, H-6 ′ose); 5.31(1H,
f
0.47 (methylene chloride/ethanol, 95/5). UV-visible
1
2H, m, H-4,5 nitrophenyl); 8.11 (6H, br d, H-2,6 tolyl); 8.28
3
, 200 MHz) δ -2.74(2H, br s,
1H, dd, J ) 7.1-2.0, H-6 nitrophenyl); 8.43 (1H, dd, J ) 7.0-
.5 Hz, H-3 nitrophenyl); 8.78 (2H, dd, J ) 4.8 Hz, H-3-7 pyr);
.86 (4H, s, H-12,13,17,18 pyr); 8.87 (2H, d, J ) 5.1 Hz, H-2,8
2
1
3
pyr). C NMR (50 MHz) 21.5 (3C, Me tolyl); 113.9 (1C, C-5
meso); 120.5 (2C, C-10,20 meso); 121.0 (1C, C-15 meso); 123.9
b
a
m, H-4′ose); 5.46(3H, m, H-1′,2′,3′ose); 7.04(6H, d, J ) 8.3 Hz,
H-3,5 aminophenyl); 7.36(2H, d, J ) 8.6 Hz, H-3,5 aryl); 7.98-
(6H, d, J ) 8.4 Hz, H-2,6 aminophenyl); 8.13(2H, d, J ) 8.5
Hz, H-2,6 aryl); 8.80(2H, d, J ) 4.8 Hz, H-3,7 â pyrrole); 8.90-
(4H, s, H-12,13,17,18 â pyrrole); 8.91(2H, d, J ) 4.7 Hz, H-2,8
(
1C, C-3 nitrophenyl); 127.4 (6C, C-3,5 tolyl); 129.4 (1C, C-4
nitrophenyl); 130.8 (1C, C-5 nitrophenyl); 131.3 (Câ pyrrole);
31.9 (Câ pyrrole); 134.5 (6C, C-2,6 tolyl); 136.8 (1C, C-1
nitrophenyl); 136.5 (1C, C-6 nitrophenyl); 137.4 (3C, C-4 tolyl);
1
pyrrole). 13C NMR (CDCl
(2C, CH
, 50 MHz) 20.7(2C, CH
3
CO); 62.1(1C, C-6′ose); 68.4(1C, C-4′ose); 71.3(1C, C-2′
1
1
39.0 (2C, C-1 tolyl); 139.2 (1C, C-1 tolyl); 146.2 (8C, CR pyr);
3
3
CO); 20.8-
52.0 (1C, C-2 nitrophenyl). MS (MALDI) m/z: 702.3 ([M +
+
H] monoisotopic) . Anal. Calcd for C47
H
35
N
5
O
2
‚H
2
O: C, 78.40;
or C-3′ose); 72.3(1C, C-5′ose); 72.9(1C, C-2′ or C-3′ose); 99.2-
(1C, C-1′ose); 113.4(6C, C-3,5 aminophenyl); 114.9(2C, C-3,5
aminophenyl); 118.7(1C, C meso); 119.1(2C, C meso); 120.0-
H, 5.18; N, 9.72. Found: C, 78.16; H, 5.15; N, 9.67.
-(4-Nitr oph en yl)-10,15,20-tr istolylpor ph yr in (8b). 4-Ni-
trobenzaldehyde (151 mg, 1 mmol, 1 equiv), p-tolualdehyde
0.35 mL, 3 mmol, 3 equiv), and pyrrole (0.28 mL, 4 mmol, 4
equiv) gave 8b 99 mg (14%). R 0.53 (methylene chloride/
petroleum ether, 60/40). UV-visible (see Table 2). H NMR
CDCl , 200 MHz) δ -2.73(2H, s, NH); 2.72(9H, s, Me tolyl);
.56(6H, d, J ) 7.8 Hz, H-3,5 tolyl); 8.10(6H, d, J ) 7.9 Hz,
H-2,6 tolyl); 8.38 (2H, d, J ) 8.7 Hz, H-2,6 nitrophenyl); 8.62
2H, d, J ) 9.0 Hz, H-3,5 nitrophenyl); 8.72(2H, d, J ) 4.8 Hz,
H-3,7 â pyrrole); 8.89 (4H, s, H-12-13-17-18 â pyrrole); 8.92
5
(
1C, C meso); 130.9(8C, C-â pyrrole); 132.6(3C, C-4 amino-
phenyl); 135.5(2C, C-2,6 aryl); 135.7(6C, C-2,6 aminophenyl);
37.6(1C, C-1 aryl); 146.2(3C, C-1 aminophenyl); 146.5(8C, C-R
pyrrole); 156.5(1C, C-4 aryl); 168.3(1C, CH CO); 169.5(1C,
CH CO); 170.4(1C, CH CO); 170.7(1C, CH CO). MS (MALDI)
m/z: 1006.4 ([M H] monoisotopic). Anal. Calcd for
10: C, 69.24; H, 5.11; N, 9.75. Found: C, 69.13; H,
.15; N, 9.76.
,15-Bis(4-a m in op h en yl)-10,20-b is[-(2′,3′,4′,6′-t et r a -O-
(
1
f
1
3
3
3
3
(
7
3
+
+
58 51 7
C H N O
5
(
5
a cetyl-â-D-glu cop yr a n osyloxy)p h en yl] P or p h yr in (10). 4
(30 mg, 0.021 mmol) and 10% Pd/C (89 mg) gave 17 mg of 10
(
2H, d, J ) 4.8 Hz, H-2-8 â pyrrole). MS (MALDI) m/z: 703
+
13
(
1
M+H) . C NMR (50 MHz) δ 149.4 (1C, C-1 nitrophenyl);
47.7 (1C, C-4 nitrophenyl); 146.2 (8C, CR pyrrole); 139.1 (3C,
C-1 tolyl); 137.5 (3C, C-4 tolyl); 135.1 (2C, C-2,6 nitrophenyl);
(
62%). R
f
0.50 (methylene chloride/ethanol, 95/5). UV-visible
1
(see Table 2). H NMR (CDCl
3
, 200 MHz) δ -2.72(2H, br s,
NH); 2.11(6H, s, OAc); 2.12(6H, s, OAc); 2.13(6H, s, OAc); 2.23-
6H, s, OAc); 3.97(4H, br s, NH
1
1
34.5 (6C, C-2,6 tolyl); 131.4 (Câ pyrrole); 130.9 (Câ pyrrole);
27.5 (6C, C-3,5 tolyl); 121.8 (2C, C-3,5 nitrophenyl); 121.0 (1C,
(
2
2
); 4.06(2H, ddd, J ) 9.6-5.3-
′ose); 4.43(2H,
a
′ose); 5.32(2H, m, H-4′ose); 5.48-
.5, H-5′ose); 4.31(2H, dd, J ) 12.3-2.0, H-6
b
C-15 meso); 120.7 (2C, C-10,20 meso); 116.8 (1C, C-15 meso);
2
topic). Anal. Calcd for C47 O: C, 74.68; H, 5.45;
N, 9.26. Found: C, 74.86; H, 5.49; N, 9.20.
+
dd, J ) 12.3-5.4 Hz, H-6
(
phenyl); 7.38(4H, d, J ) 8.6 Hz, H-3,5 aryl); 7.98(4H, d, J )
8
8
4
1.5 (3C, Me tolyl). MS (MALDI) m/z: 702.3 ([M + H] monoiso-
6H, m, H-1′,2′,3′ose); 7.03(4H, d, J ) 8.4 Hz, H-3,5 amino-
H
35
N
5
O
2
‚3H
2
.3 Hz, H-2,6 aminophenyl); 8.14(4H, d, J ) 8.6 Hz, H-2,6 aryl);
Gen er a l P r oced u r e for R ed u ct ion of Glu cosyla t ed
.83(4H, d, J ) 4.8 Hz, H-2,8,12,18 â pyrrole); 8.93(4H, d, J )
Nitr op h en ylp or p h yr in s 3-6. Porphyrins 3a ,b, 4, 5, and
6
added. The mixture was stirred under H for 5 h, and then
2
the reaction was filtered through Celite. The solvent was
removed in vacuo, and the pure product was obtained after
purification on thin-layer chromatography.
13
.8 Hz, H-3,7,13,17 â pyrrole). C NMR (CDCl
3
, 50 MHz) 20.6-
a ,b were dissolved in THF and 10% palladium on carbon was
(
2C, CH
C-5′ose); 71.4(2C, C-2′ or C-3′ose); 72.3(2C, C-2′ or C-3′ose);72.9-
2C, C-2′ or C-3′ose); 99.2(2C, C-1′ose); 113.4(4C, C-3,5 ami-
3 3
CO); 20.8(2C, CH CO); 62.1(2C, C-6′ose); 68.5(2C,
1
3
(
nophenyl); 115.0(4C, C-3,5 aryl); 118.7(2C, C meso); 120.5(2C,
C meso); 130.7(C-â pyrrole); 131.2(C-â pyrrole); 132.3(2C, C-4
aminophenyl); 135.5(4C, C-2,6 aryl); 135.6(4C, C-2,6 ami-
nophenyl); 137.4(2C, C-1 aryl); 146.0(2C, C-1 aminophenyl);
5
-[2-(2′,3′,4′,6′-Tetr a -O-a cetyl-â-D-glu cop yr a n osyloxy)-
p h en yl]-10,15,20-tr is(4-a m in op h en yl) P or p h yr in (9a ). 3a
(
(
2
52 mg, 0.048 mmol) and 10% Pd/C (300 mg) gave 28 mg of 9a
60%). R 0.26 (toluene/acetone, 60/40). UV-visible (see Table
). H NMR (CDCl , 200 MHz) δ -2.74(2H, br s, NH); 0.92-
3H, s, OAc); 1.26(3H, s, OAc); 1.86(3H, OAc); 1.96(3H, OAc);
146.5(8C, C-R pyrrole); 156.6(2C, C-4 aryl); 169.4(4C, CH CO);
3
f
170.3(2C, CH CO); 170.6(2C, CH CO). MS (MALDI) m/z:
3
3
1
+
3
1337.6 ([M + H] monoisotopic). Anal. Calcd for C H N O :
7
2
68
6
20
(
C, 64.67; H, 5.12; N, 6.28. Found: C, 64.91; H, 5.14; N, 6.31.
5,10-Bis(4-a m in op h en yl)-15,20-b is[-(2′,3′,4′,6′-t et r a -O-
a cetyl-â-D-glu cop yr a n osyloxy)p h en yl] P or p h yr in (11). 5
(30 mg, 0.021 mmol) and 10% Pd/C (89 mg) gave 18 mg of 11
f
3
1
1
4
.65(1H, ddd, J ) 7.7-5.1-2.5 Hz, H-5′ ose); 4.00(1H, dd, J )
2.1-2.6 Hz, H-6 ′ose); 4.03(2H, br s, NH ); 4.12(1H, dd, J )
2.1-5.2 Hz, H-6 ′ose); 4.18(1H, dd, J ) 7.7-2.2 Hz, H-2′ose);
.64(1H, t, J ) 9.2 Hz, H-3′ose); 4.70(1H, t, J ) 9.2 Hz,
b
2
a
(64%). R 0.55 (methylene chloride/ethanol, 95/5). UV-visible
1
H-4′ose); 4.92(1H, d, J ) 7.7 Hz, H-1′ose); 7.04(6H, d, J ) 8.3
Hz, H-3,5 aminophenyl); 7.52(1H, dt, J ) 7.5-1.6 Hz, H-5
aryl); 7.70(1H, dt, J ) 7.5-1.6 Hz, H-3 aryl); 7.80(1H, dt, J )
(see Table 2). H NMR (CDCl , 200 MHz) -2.71(2H, br s, NH);
3
2.11(6H, s, OAc); 2.12(6H, s, OAc); 2.13(6H, s, OAc); 2.23(6H,
s, OAc); 3.97(4H, br s, NH ); 4.06(2H, ddd, J ) 9.6-5.3-2.4,
2
7
.5-1.6 Hz, H-4 aryl); 7.98(6H, d, J ) 8.4 Hz, H-2,6 ami-
H-5′ose); 4.31(2H, dd, J ) 12.2-2.4, H-6 ′ose); 4.44(2H, dd, J
b
nophenyl); 8.85(2H, d, J ) 4.9 Hz, H-3,7 â pyrrole); 8.90(4H,
s, H-12,13,17,18 â pyrrole); 8.92(2H, d, J ) 4.9 Hz, H-2,8 â
) 12.3-5.4, H-6 ′ose); 5.33(2H, m, H-4′ose); 5.48(6H, m,
a
H-1′,2′,3′ose); 7.02(4H, d, J ) 8.3 Hz, H-3.5 aminophenyl); 7.39-
(4H, d, J ) 8.6 Hz, H-3.5 aryl); 7.98(4H, d, J ) 8.3 Hz, H-2,6
aminophenyl); 8.15(4H, d, J ) 8.6 Hz, H-2,6 aryl); 8.83(2H, d,
J ) 4.9 Hz, H-2,13 â pyrrole); 8.84(2H, s, H-17,18 â pyrrole);
8.93(2H, s, H-7,8 â pyrrole); 8.94(2H, d, J ) 4.7 Hz, H-3,12 â
1
3
pyrrole). C NMR (CDCl
1C, CH CO); 20.5(2C, CH
C-4′ ose); 70.1(1C, C-2′ ose); 72.0(1C, C-5′ose); 72.3(1C, C-3′ose);
9.7(1C, C-1′ose); 113.4(6C, C-3,5 aminophenyl); 116.4(1C, C
3
, 50 MHz) 17.5(1C, CH
3
CO); 20.3-
(
3
3
CO); 62.1(1C, C-6′ ose); 68.4(1C,
9
pyrrole). 13C NMR (CDCl
(2C, CH
, 50 MHz) 20.6(2C, CH
CO); 62.1(2C, C-6′ose); 68.5(2C, C-5′ose); 71.4(2C, C-2′
meso); 117.8(1C, C-2 aryl); 119.7(2C, C meso); 120.1(1C, C
meso); 122.7(1C, C-5 aryl); 130.4(1C, C-4 aryl); 131.0(8C, C-â
pyrrole); 132.2(1C, C-1 aryl); 132.6(3C, C-4 aminophenyl);
3
3
CO); 20.7-
3
or C-3′ose); 72.3(2C, C-2′ or C-3′ose); 72.9(2C, C-2′ or C-3′ose);
99.2(2C, C-1′ose); 113.4(4C, C-3,5 aminophenyl); 115.1(4C,
C-3,5 aryl); 118.6(2C, C meso); 120.8(2C, C meso); 130.7(C-â
pyrrole); 131.2(C-â pyrrole); 132.4(2C, C-4 aminophenyl);
135.5(4C, C-2,6 aryl); 135.6(4C, C-2,6 aminophenyl); 137.4-
(2C, C-1 aryl); 146.0(2C, C-1 aminophenyl); 146.5(8C, C-R
1
35.7(6C, C-2,6 aminophenyl); 135.9(1C, C-6 aryl); 146.0(3C,
C-1 aminophenyl); 146.5(8C, C-R aminophenyl); 156.2(1C, C-2
aryl); 166.9(1C, CH CO); 169.0(1C, CH CO); 169.5(1C, CH CO);
70.3(1C, CH CO). MS (MALDI) m/z: 1006.4 ([M + H]
monoisotopic). Anal. Calcd for C58 10: C, 69.24; H, 5.11;
N, 9.75. Found: C, 69.04; H, 5.08; N, 9.72.
3
3
3
+
1
3
51 7
H N O
3
pyrrole); 156.6(2c, C-4 aryl); 169.4(4C, CH CO); 170.3(2C,

Glycosylated Amino Acid Porphyrin Derivatives
J . Org. Chem., Vol. 64, No. 12, 1999 4441
CH
3
+
CO); 170.6(2C, CH
3
CO). MS (MALDI) m/z: 1337.6 ([M +
H] monoisotopic). Anal. Calcd for C72 20: C, 64.67; H,
.12; N, 6.28. Found: C, 64.82; H, 5.15; N, 6.30.
-(2-Am in op h en yl)-10,15,20-t r is[4-(2′,3′,4′,6′-t et r a -O-
a cetyl-â-D-glu cop yr a n osyloxy)p h en yl] P or p h yr in (12a ).
8.11(6H, d, J ) 8.0 Hz, H-2,6 tolyl); 8.88(2H, d, J ) 4.7 Hz,
H-2,8 â pyrrole); 8.89(4H, s, H-12,13,17,18 â pyrrole); 8.90-
68 6
H N O
5
(2H, d, J ) 4.7 Hz, H-3,7 â pyrrole). 13C NMR (CDCl
3
, 50 MHz)
5
146.7 (1C, C-1 aminophenyl); 146.2 (8C, CR pyr); 139.3 (1C,
C-1 tolyl); 139.1 (2C, C-1 tolyl); 137.4 (3C, C-4 tolyl); 134.8
(1C, C-1 aminophenyl); 134.5 (6C, C-2,6 tolyl); 131.7(Câ pyr);
130.9 (Câ pyr); 129.5 (1C, C-3 aminophenyl); 127.5 (1C, C-5
aminophenyl); 127.4 (6C, C-3,5 tolyl); 120.7 (1C, C-20 meso);
120.1 (2C, C-10,15 meso); 117.5 (1C, C-6 aminophenyl); 115.5
(1C, C-2 aminophenyl); 115.1 (1C, C-5 meso); 21.5 (3C, Me
6
1
a (67 mg, 0.04 mmol) and 10% Pd/C (85 mg) gave 40 mg of
2a (60%). R 0.40 (toluene/acetone, 70/30). UV-visible (see
, 200 MHz) δ -2.75(2H, br s, NH);
.11(18H, s, OAc); 2.12(12H, s, OAc); 2.22(6H, s, OAC); 3.55-
2H, br s, NH ); 4.07(3H, ddd, J ) 9.6-5.4-2.5 Hz, H-5′ose);
.31(3H, dd, J ) 12.3-2.0 Hz, H-6 ′ose); 4.43(3H, dd, J )
2.3-5.3 Hz, H-6 ′ose); 5.31(3H, m, H-4′ose); 5.47(9H, m,
f
1
Table 2). H NMR (CDCl
2
(
4
1
3
2
+
b
tolyl). MS (MALDI) m/z: 672.3 ([M + H] monoisotopic). Anal.
a
Calcd for C47
C, 79.49; H, 5.87; N, 9.93.
H
37
N
5
2
‚2H O: C, 79.75; H, 5.84; N, 9.89. Found:
H-1′,2′,3′ose); 7.15(2H, m, H-5,6 o-aminophenyl); 7.39(6H, d,
J ) 8.6 Hz, H-3,5 aryl); 7.60(1H, m, H-4 o-aminophenyl); 7.87-
5-(4-Am in op h en yl)-10,15,20-t r ist olylp or p h yr in (13b):
1
(
8
1H, d, J ) 7.6-1.3 Hz, H-3 o-aminophenyl); 8.13(6H, d, J )
43.2 mg (90%); R
NMR (CDCl
f
0.34 (CH
2
Cl
2
). UV-visible (see Table 2). H
.5 Hz, H-2,6 aryl); 8.85(4H, s,H-12,13,17,18 â pyrrole); 8.86-
3
, 200 MHz) δ 8.87(d, J ) 4.9 Hz, 2H, H-3,7 pyr);
(2H, d, J ) 4.7 Hz, H-2,8 â pyrrole); 8.91(2H, d, J ) 8.91 Hz,
8.85(s, 4H, H-12,13,17,18 pyr); 8.83(d, J ) 4.9 Hz, 2H, H-2,8);
8.10(d, J ) 7.9 Hz, 6H, H-2,6 tolyl); 7.90(d, J ) 8.3 Hz, 2H,
H-2,6 aminophenyl); 7.56(d, J ) 7.9 Hz, 6H, H-3,5 tolyl); 6.72-
1
3
H-3,7 â pyrrole). C NMR (CDCl
0.8(6C, CH CO), 62.1(3C, C-6′ose); 68.5(3C, C-4′ose); 71.4-
3C, C-2′ or C-3′ose); 72.3(3C, C-5′ose); 72.9(3C, C-2′ or C-3′ose);
3 3
, 50 MHz) 20.6(6C, CH CO);
2
(
3
(d, J ) 8.3 Hz, 2H, H-3,5 aminophenyl); 3.45(bs, 2H, NH
2
);
, 50
9
1
1
9.2(3C, C-1′ose); 115.1(6C, C-3,5 aryl); 115.1(1C, C meso);
15.7(1C, C-2 aminophenyl); 117.6(1C, C-6 aminophenyl);
19.2(3C, C meso); 127.1(1C, C-5 aminophenyl); 129.7(1C, C-3
2.71(s, 9H, Me tolyl); -2.73(s, 2H, NH). 13C NMR (CDCl
3
MHz) δ 146.2 (8C, CR pyr); 146.1 (1C, C-1 aminophenyl); 139.4
(3C, C-1 tolyl); 137.2 (3C, C-4 tolyl); 135.4 (2C, C-2,6 ami-
nophenyl); 134.5 (6C, C-2,6 tolyl); 132.7 (1C, C-4 aminophenyl);
131.7(C-â pyr); 130.8 (C-â pyr); 127.4 (6C, C-3,5 tolyl); 119.9
(3C, C-10,15,20 meso); 119.1 (1C, C-5 meso); 113.4 (2C, C-3,5
aminophenyl); 130.9(C-â pyrrole); 134.8(1C, C-4 aminophenyl);
1
1
35.5(6C, C-2,6 aryl); 137.0(4C, C-1 aryl); 137.2(2C, C-1 aryl);
46.5(8C, C-R pyrrole); 146.9(1C, C-1 aminophenyl); 156.7(3C,
C-4 aryl); 169.4(6C, CH
CH
Anal. Calcd for C86
C, 61.87; H, 5.15; N, 4.18.
-(4-Am in op h en yl)-10,15,20-t r is[4-(2′,3′,4′,6′-t et r a -O-
a cetyl-â-D-glu cop yr a n osyloxy)p h en yl] P or p h yr in (12b).
3
CO); 170.3(3C, CH
3
+
CO); 170.6(3C,
aminophenyl); 21.5 (3C, Me tolyl). MS (MALDI) m/z: 672.3 ([M
+
3
CO). MS (MALDI) m/z: 1668.5 ([M + H] monoisotopic).
+ H] monoisotopic). Anal. Calcd for C47
H
37
N
5
2
‚3H O: C, 77.76;
H
85
N
5
O
30: C, 61.90; H,5.13; N, 4.20. Found:
H, 5.96; N, 9.65; Found: C, 77.54; H, 5.97; N, 9.69.
Gen er a l P r oced u r e for Bin d in g Am in o Acid s on P or -
p h yr in s Am in o F u n ction . Porphyrins 9a ,b, 10, 11, 12a ,b,
5
and 13a ,b were reacted with Fmoc amino acid (3 equiv/NH
in the presence of dicyclohexylcarbodiimide (DCC) (3 equiv/
NH ) in CH Cl (10 mL). The mixture was stirred in darkness
2
)
6
1
b (25 mg, 0.015 mmol) and 10% Pd/C (32 mg) gave 19 mg of
2b (76%). R 0.44 (methylene chloride/ethanol, 90/10). UV-
f
2
2
2
1
visible (see Table 2). H NMR (CDCl
3
, 200 MHz) δ -2.75(2H,
at room temperature for 15 h. The solution was concentrated
in vacuo, and ethyl acetate was added to the residue in order
to remove insoluble dicyclohexylurea by filtration. The solvent
was evaporated, and the residue was purified on TLC.
5-[2-(2′,3′,4′,6′-Tetr a -O-a cetyl-â-D-glu cop yr a n osyloxy)-
p h en yl]-10,15,20-tr is[4-(N-(9-flu or en ylm eth oxyca r bon yl)-
L-a la n yla m in o)p h en yl] P or p h yr in (14a ). 9a (30 mg, 0.03
mmol), Fmoc-L-alanine (84 mg, 0.27 mmol), and dicyclohexy-
br s, NH); 2.11(9H, s, OAc); 2.12(9H, s, OAc); 2.13(9H, s, OAc);
2
5
4
5
2
.23(9H, s, OAc); 4.00(2H, br s, NH ); 4.07(3H, ddd, J ) 9.6-
.4-2.5 Hz, H-5′ose); 4.31(3H, dd, J ) 12.2-2.5 Hz, H-6 ′ose);
.44(3H, dd, J ) 12.2-5.3 Hz, H-6 ′ose); 5.35(3H, m, H-4′ose);
.49(9H, m, H-1′,2′,3′ ose); 7.05(2h, d, J ) 8.4 Hz, H-3,5
b
a
aminophenyl); 7.39(6H, d, J ) 8.5 Hz, H-3,5 aryl); 7.98(2H, d,
J ) 8.3 Hz, H-2,6 aminophenyl); 8.14(6H, d, J ) 8.6 Hz, H-2,6
aminophenyl); 8.83(2H, d, J ) 4.9 Hz, H-2,8 â pyrrole); 8.85-
lcarbodiimide (54 mg, 0.27 mmol) gave 51 mg of 14a (90%). R
f
1
(4H, s, H-12,13,17,18 â pyrrole); 8.95(2H, d, J ) 4.9 Hz, H-3,7
0.50 (toluene/acetone, 70/30). UV-visible (see Table 2).
NMR (CDCl , 400 MHz) δ -2.81(2H, br s, NH); -1.11(3H, s,
OAc); 1.28(3H, s, OAc); 1.64(9H, d, J ) 6.9 Hz, CH alanine);
1.84(3H, OAc); 2.02(3H, OAc); 3.65(1H, ddd, J ) 7.7-5.1-2.5
Hz, H-5′ose); 4.01(1H, br d, J ) 10.1 Hz, H-6 ′ose); 4.12(1H,
dd, J ) 12.1-5.1 Hz, H-6 ′ose); 4.20(1H, dd, J ) 7.9-1.5 Hz,
H-2′ose); 4.29(3H, t, J ) 7.2 Hz, H-9 Fmoc); 4.54(9H, m, CH
H
1
3
â pyrrole). C NMR (CDCl
6C, CH CO), 62.1(3C, C-6′ose); 68.4(3C, C-4′ose); 71.4(3C, C-2′
or C-3′ose); 72.3(3C, C-5′ose); 72.9(3C, C-2′ or C-3′ose); 99.2-
3C, C-1′ose); 113.2(2C, C-3,5 aminophenyl); 115.1(6C, C-3,5
aryl); 118.8(1C, C meso); 119.1(2C, C meso); 119.3(1C, C meso);
30.9(C-â pyrrole); 132.2(1C, C-4 aminophenyl); 135.0(2C,
C-2,6 aminophenyl); 135.5(6C, C-2,6 aryl); 137.2(3C, C-1 aryl);
46.1(1C, C-1 aminophenyl); 146.5(8C, C-R pyrrole); 156.6(3C,
C-4 aryl); 169.4(6C, CH CO); 170.3(3C, CH CO); 170.5(3C,
CH CO), MS (MALDI) m/z: 1668.5 ([M + H] monoisotopic).
Anal. Calcd for C86 30: C, 61.90; H, 5.13; N, 4.20. Found:
C, 61.86; H, 5.10; N, 4.22.
3
, 50 MHz) 20.6(6C, CH
3
CO); 20.7-
3
(
3
3
(
b
a
1
2
Fmoc); 4.56(1H, t, J ) 9.4 Hz, H-3′ose); 4.67(3H, CH alanine);
4.71(1H, t, J ) 9.5 Hz, H-4′ose); 4.83(1H, d, J ) 7.8 Hz,
H-1′ose); 5.96(3H, m, NH alanine); 7.32(6H, t, J ) 7.5 Hz,
H-Fmoc); 7.37(6H, t, J ) 7.7 Hz, H-Fmoc); 7.64(7H, m,
H-Fmoc and H-3 aryl); 7.71(7H, m, H-5 aryl and H-Fmoc);
7.87(6H, br d, J ) 8.1 Hz, H-3,5 amidophenyl); 7.91(1H, br d,
J ) 6.4 Hz, H-4 aryl); 8.05(6H, br d, J ) 8.3 Hz, H-2,6
amidophenyl); 8.16(1H, br d, J ) 7.37 Hz, H-6 aryl); 8.65(2H,
d, J ) 4.8 Hz, H-2,8 â pyrrole); 8.74(2H, d, J ) 4.8 Hz, H-3,7
â pyrrole); 8.79(4H, s, H-12,13,17,18 â pyrrole); 8.91(3H, br s,
1
3
3
+
3
85 5
H N O
Gen er a l P r oced u r e for Red u ction of Mon on itr op h en -
yltr istolylp or p h yr in . Reductions were performed according
to methods already described in the literature. Mononitro-
phenyltristolylporphyrin 8a ,b was dissolved in 9 mL of
1
4
chloroform/acetic acid (1/2), and then a solution (3 mL) of SnCl
3 equiv/NO ) in concentrated HCl was added. The mixture
2
NH amidophenyl). 13C NMR (CDCl
CO); 18.6(3C, CH alanine); 19.9(1C, CH
CO), 20.7(1C, CH
alanine); 61.9(1C, C-6′ ose); 67.5(3C, CH
3
, 100 MHz) 17.4(1C, CH
CO); 20.4(1C, CH
3
-
-
(
2
3
3
3
was stirred, refluxed overnight, and then neutralized with 1
M sodium hydroxide to pH 8-9, an ice-bath being used for
cooling. Chloroform (250 mL) was added, and the organic layer
was washed with water (3 × 75 mL), followed by drying over
magnesium sulfate. The solvent was evaporated in a vacuum.
The crude product was purified on TLC (CH
5
3
CO); 47.2(3C, CH-9 Fmoc); 49.2(3C, CH
Fmoc); 68.1(1C, C-4′
2
ose); 70.1(1C, C-2′ ose); 71.8(1C, C-5′ose); 72.1(1C, C-3′ose);
99.6(1C, C-1′ose); 114.7(1C, C meso); 118.2(7C, C-3 aryl and
C-3,5 amidophenyl); 119.2(1C, C meso); 119.6(1C, C meso);
119.8(1C, C meso); 120.1(6C, C-Fmoc); 122.5(1C, C-5 aryl);
125.0(6C, C-Fmoc); 127.2(6C, C-Fmoc); 127.6(6C, C-Fmoc);
130.7(1C, C-4 aryl); 131.0(8C, C-â pyrrole); 133.0(1C, C-1 aryl);
135.1(6C, C-2,6 amidophenyl); 136.1(1C, C-6 aryl); 137.5(3C,
2
Cl
2
).
1
5
-(2-Am in op h en yl)-10,15,20-tr istolylp or p h yr in (13a ):
1
4
f 2 2
3 mg (92%). R 0.52 (CH Cl ). UV-visible (see Table 2). H
NMR (CDCl
tolyl); 3.50(2H, br s, NH
aminophenyl); 7.18(1H, dt, J ) 7.4-1.0 Hz, H-5 aminophenyl);
.56(6H, d, J ) 8.0 Hz, H-3,5 tolyl); 7.61(1H, m, H-4 ami-
nophenyl); 7.91(1H, dd, J ) 7.5-1.5 Hz, H-3 aminophenyl);
3
, 200 MHz) δ -2.70(2H, s, NH); 2.72(9H, s, Me
IV
2
); 7.07(1H, dd, J ) 8.0-0.9 Hz, H-6
C-1 amidophenyl); 138.2(3C, C-4 amidophenyl); 141.3(6C, C
IV
Fmoc); 143.6(6C, C Fmoc); 146.0(4C, C-R Pyrrole); 156.2(1C,
7
C-2 aryl); 156.6(3C, CO Fmoc); 167.0(1C, CH
3
CO); 169.1(1C,
CH CO); 169.5(1C, CH CO); 170.5(1C, CH CO); 171.1(3C, CO
3 3 3

4
442 J . Org. Chem., Vol. 64, No. 12, 1999
Sol et al.
+
alanine). MS (MALDI) m/z: 1885.7 ([M + H] monoisotopic).
Anal. Calcd for C112 O: C, 69.98; H, 5.24; N, 7.29.
Found: C, 70.08; H, 5.27; N, 7.34.
-[4-(2′,3′,4′,6′-Tetr a -O-a cetyl-â-D-glu cop yr a n osyloxy)-
Anal. Calcd for C108
Found: C, 66.10; H, 5.21; N, 5.67.
5,10-Bis[4-(N-(9-flu or en ylm eth oxyca r bon yl)-L-a la n yl-
amino)phenyl]-15,20-bis[4-(2′,3′,4′,6′-tetra-O-acetyl-â-D-gluco-
p yr a n osyloxy)p h en yl] P or p h yr in (16). 11 (41 mg, 0.03
mmol), Fmoc-L-alanine (56 mg, 0.18 mmol), and dicyclohexy-
H
98
N
8
O
26‚2H
2
O: C, 66.18; H, 5.24; N, 5.71.
H
96
N
10
O
19‚2H
2
5
ph en yl]-10,15,20-tr is[4-(N-(9-flu or en ylm eth oxyca r bon yl)-
L-a la n yla m in o)p h en yl] P or p h yr in (14b). 9b (15 mg, 0.015
mmol), Fmoc-L-alanine (42 mg, 0.14 mmol), and dicyclohexy-
lcarbodiimide (37 mg, 0.18 mmol) gave 48 mg of 16 (85%). R
f
1
lcarbodiimide (28 mg, 0.14 mmol) gave 51 mg of 14b (89%). R
f
0.45 (toluene/acetone, 60/40). UV-visible (see Table 2).
NMR (CDCl , 200 MHz) δ -2.87(2H, br s, NH); 1.64(6H, d, J
) 6.9 Hz, CH alanine); 2.08(6H, s, OAc); 2.10(6H, s, OAc);
H
0
2
.42 (methylene chloride/ethanol, 95/5). UV-visible (see Table
3
1
). H NMR (CDCl
3
, 300 MHz) δ -2.84(2H, br s, NH); 1.65-
alanine); 2.10(3H, s, OAc); 2.11(3H, s,
3
(9H, d, J ) 7.1 Hz, CH
3
2.11(6H, s, OAc); 2.21(6H, s, OAc); 4.06(2H, ddd, J ) 9.6-
5.2-2.5, H-5′ose); 4.29(2H, m, H-9 Fmoc); 4.31(2H, br dd, J )
OAc); 2.11(3H, br s, OAc); 2.21(3H, br s, OAc); 4.06(1H, ddd,
J ) 9.6-5.2-2.5 Hz, H-5′ose); 4.30(1H, dd, J ) 12.1-2.6 Hz,
10.2-2.6, H-6
b
′ose); 4.43(2H, dd, J ) 12.1-5.3 Hz, H-6
a
′ose);
H-6
b
′ose); 4.42(1H, dd, J ) 12.1-5.2 Hz, H-6
a
′ose); 4.18(1H,
4.58(6H, br d, J ) 6.8 Hz, CH
2
Fmoc and CH alanine); 5.30-
dd, J ) 7.7-2.2 Hz, H-2′ose); 4.28(3H, m, H-9 Fmoc); 4.56-
3H, d, J ) 6.8 Hz, CH Fmoc); 4.58(3H, d, J ) 6.8 Hz, CH
alanine); 5.31(1H, m, H-4′ose); 5.48(3H, m, H-1′,2′,3′ose); 5.65-
3H, m, NH alanine); 7.29(3H, dt, J ) 7.5-2.4 Hz, H-Fmoc);
.35(2H, d, J ) 8.6 Hz, H-3,5 aryl); 7.38(3H, dt, J ) 7.5-2.4
Hz, H-Fmoc); 7.66(3H, dt, J ) 7.5-2.4 Hz, H-Fmoc); 7.70-
1H, dt, J ) 7.5-1.6 Hz, H-3 aryl); 7.75(3H, d, J ) 7.5 Hz,
(2H, br t, J ) 9.6 Hz, H-4′ose); 5.49(6H, m, H-1′,2′,3′ose); 5.60-
(2H, m, NH alanine); 7.26-7.35(8H, m, H-Fmoc); 7.38(4H,br
d, J ) 8.6 Hz, H-3,5 aryl); 7.65(4H, br dd, J ) 6.8-2.7 Hz,
H-Fmoc); 7.76(4H, d, J ) 6.8 Hz, H-Fmoc); 7.90(4H, d, J )
8.6 Hz, H-3,5 amidophenyl); 8.15(8H, d, J ) 8.6 Hz, H-2,6
amidophenyl and H-2,6 aryl); 8.70(2H, br s, NH-Fmoc); 8.81-
(2H, d, J ) 4.8 Hz, H-3,12 â pyrrole); 8.83(4H, s, H-7,8,17,18
(
2
(
7
(
1
3
H-Fmoc); 7.80(1H, dt, J ) 7.5-1.6 Hz, H-4 aryl); 7.91(6H, br
d, J ) 8.6 Hz, H-3,5 amidophenyl); 8.11(2H, br d, J ) 8.6 Hz,
H-2,6 aryl); 8.15(6H, br d, J ) 8.6 Hz, H-2,6 amidophenyl);
â pyrrole); 8.84(2H, d, J ) 4.8 Hz, H-2,13 â pyrrole). C NMR
(CDCl , 50 MHz) 18.4(2C, CH alanine); 20.7(4C, CH CO);
20.8(4C, CH CO); 47.2(2C, CH-9 Fmoc); 49.2(2C, CH alanine);
62.1(2C, C-6′ ose); 67.4(2C, CH Fmoc); 68.4(2C, C-4′ ose); 71.3-
3
3
3
3
8
.70(3H, br s, NH amidophenyl); 8.80(2H, d, J ) 4.8 Hz, H-3,7
â pyrrole); 8.84(4H, s, H-12,13,17,18 â pyrrole); 8.87(2H, d, J
4.8 Hz, H-2,8 â pyrrole). 13C NMR (CDCl
, 100 MHz) 18.3-
3C, CH alanine); 20.6(1C, CH CO); 20.7(1C, CH CO); 47.2-
3C, CH-9 Fmoc); 51.4(3C, CH alanine); 62.1(1C, C-6′ ose);
7.4(3C, CH Fmoc); 68.5(1C, C-4′ ose); 71.4(1C, C-2′ or C-3′
2
(2C, C-2′ or C-3′ ose); 72.3(2C, C-5′ose); 72.9(2C,C-2 or C-3′ose);
99.1(2C, C-1′ose); 114.9(4C, C-3,5 aryl); 118.2(4C, C-3,5 ami-
dophenyl); 119.2(2C, C meso); 119.5(2C, C meso); 120.1(4C,
C-Fmoc); 125.0(4C, C-Fmoc); 127.2(4C, C-Fmoc); 127.9(4C,
C-Fmoc); 130.9(8C, C-â pyrrole); 135.1(4C, C-2,6 amido-
phenyl); 135.4(4C, C-2,6 aryl); 137.1(2C, C-1 aryl); 137.4(2C,
)
(
(
3
3
3
3
6
2
ose); 72.3(1C, C-5′ose); 72.9(1C, C-3′ or C-2′ ose); 99.1(1C,
C-1′ose); 115.1(2C, C-3,5 aryl); 118.2(6C, C-3,5 amidophenyl);
IV
C-1 amidophenyl); 138.2(2C, C-4 amidophenyl); 141.4(4C, C
IV
1
1
1
19.4(1C, C meso); 119.7(3C, C meso); 120.1(6C, C-Fmoc);
24.9(6C, C-Fmoc); 127.2(6C, C-Fmoc); 127.7(6C, C-Fmoc);
31.0(8C, C-â pyrrole); 135.0(6C, C-2,6 amidophenyl); 135.5-
Fmoc); 143.7(4C, C Fmoc); 146.2(4C, C-R Pyrrole); 156.6(2C,
C-4 aryl); 156.7(2C, CO Fmoc); 169.5(4C, CH CO); 170.4(2C,
3
CH
DI) m/z: 1923.7 ([M + H] monoisotopic). Anal. Calcd for
O: C, 65.58; H, 5.30; N, 5.66. Found: C,
3 3
CO); 170.7(2C, CH CO); 170.8(2C, CO alanine). MS (MAL-
+
(
2C, C-2,6 aryl); 137.1(1C, C-1 aryl); 137.4(3C, C-1 amido-
IV
phenyl); 138.2(3C, C-4 amidophenyl); 141.3(6C, C Fmoc);
C
108
H
98
N
8
O
26‚3H
2
IV
1
43.7(6C, C Fmoc); 146.2(4C, C-R pyrrole); 156.7(4C, CO
Fmoc and C-4 aryl); 169.5(2C, CH CO); 170.4(2C, CH CO);
70.7(3C, CO alanine). MS (MALDI) m/z: 1885.7 ([M +
65.82; H, 5.31; N, 5.65.
3
3
5-[2-(N-9-F lu or en ylm eth oxyca r bon yl)-L-a la n yla m in o)-
p h en yl]-10,15,20-t r is[4-(2′,3′,4′,6′-t et r a -O-a cet yl-â-D-glu -
cop yr a n osyloxy)p h en yl] P or p h yr in (17a ). 12a (15 mg,
0.009 mmol), Fmoc-L-alanine (9 mg, 0.027 mmol), and dicy-
clohexylcarbodiimide (6 mg, 0.0027 mmol) gave 15 mg of 17a
1
+
H] monoistopic). Anal. Calcd for C112
.13; N, 7.43. Found: C, 71.46; H, 5.15; N, 7.47.
,15-Bis[4-(N-(9-flu or en ylm eth oxyca r bon yl)-L-a la n yl-
96 10
H N O19: C, 71.32; H,
5
5
amino)phenyl]-10,20-bis[4-(2′,3′,4′,6′-tetra-O-acetyl-â-D-gluco-
p yr a n osyloxy)p h en yl] P or p h yr in (15). 10 (50 mg, 0.037
mmol), Fmoc-L-alanine (69 mg, 0.22 mmol), and dicyclohexy-
(85%). R
f
0.34 (toluene/acetone, 70/30). UV-visible (see Table
2). H NMR (CDCl , 200 MHz) δ -2.73(2H, br s, NH); 0.51-
(3H, br s, CH alanine); 2.11(12H, s, OAc); 2.12(12H, s, OAc);
2.22(12H, s, OAC); 3.22(1H, m, H-9 Fmoc); 3.38(2H, br s, NH );
3.50(2H, m, CH Fmoc); 4.06(3H, ddd, J ) 9.6-5.1-2.5 Hz,
H-5′ose); 4.31(4H, m, H-6 ′ose and NH alanine); 4.44(3H, dd,
J ) 12.3-5.3 Hz, H-6 ′ose); 5.32(3H, m, H-4′ose); 5.48(9H, m,
1
3
3
lcarbodiimide (46 mg, 0.22 mmol) gave 63 mg of 15 (88%). R
.47 (toluene/acetone, 60/40). UV-visible (see Table 2). 1
NMR (CDCl , 400 MHz) δ -2.80(2H, br s, NH); 1.64(6H, d, J
6.9 Hz, CH alanine); 2.11(6H, s, OAc); 2.12(12H, s, OAc);
f
2
0
H
2
3
b
)
3
a
2
4
5
.23(6H, s, OAc); 4.07(2H, ddd, J ) 9.6-5.2-2.5, H-5′ose);
H-1′,2′,3′ose); 6.21(2H, dt, J ) 7.6-1.2 Hz, H-Fmoc); 6.30-
(2H, dt, J ) 7.6-1.2 Hz, H-Fmoc); 6.41(2H, br d, J ) 7.4 Hz,
H-Fmoc); 6.68(2H, br t, J ) 7.3 Hz, H-Fmoc); 7.37(6H, d, J
) 8.1 Hz, H-3,5 aryl); 7.55(1H, dt, J ) 7.6-1.1 Hz, H-5
o-aminophenyl); 7.86(1H, dt, J ) 7.6-1.2 Hz, H-4 o-amino-
phenyl); 8.02(1H, dd, J ) 7.6-1.2 Hz, H-6 o-aminophenyl);
8.11(6H, d, J ) 8.2 Hz, H-2,6 aryl); 8.68(1H, d, J ) 8.1 Hz,
H-3 o-aminophenyl); 8.77(2H, d, J ) 4.7 Hz, H-2,8 or H-3,7 â
pyrrole); 8.82(4H, s,H-12,13,17,18 â pyrrole); 8.87(2H, d, J )
8.91 Hz, H-3,7 or H-2,8 â pyrrole); 8.87(1H, br s, NH ami-
.29(2H, m, H-9 Fmoc and H-6
.2, H-6 ′ose); 4.58(4H, m, CH
b
′ose); 4.43(2H, dd, J ) 12.2-
alanine); 4.62(2H, br s, CH
a
2
alanine); 5.31(2H, m, H-4′ose); 5.49(6H, m, H-1′,2′,3′ose); 5.61-
2H, m, NH-alanine); 7.30(4H, dt, J ) 7.0-3.1 Hz, H-Fmoc);
.40(4H, dt, J ) 7.0-3.0 Hz, H-Fmoc); 7.38(4H,br d, J ) 8.6
Hz, H-3.5 aryl); 7.65(4H, dd, J ) 8.2-3.0 Hz, H-Fmoc); 7.77-
(
7
(
4H, m, H-Fmoc); 7.95(4H,br d, J ) 8.5 Hz, H-3,5 amido-
phenyl); 8.15(8H,br d, J ) 8.6 Hz, H-2,6 aryl and H-2,6
amidophenyl); 8.63(2H, br s, NH amidophenyl); 8.84(4H, d, J
)
4.8 Hz, H-3,7,13,17 â pyrrole); 8.87(4H, d, J ) 4.8 Hz, H-2,8,-
dophenyl). 13C NMR (CDCl
20.6(6C, CH CO); 20.7(6C, CH
50.1(1C, CH alanine); 62.1(3C, C-6′ ose); 65.5(1C, CH
2
3
, 50 MHz) 17.8(1C, CH
CO); 45.7(1C, CH-9 Fmoc);
Fmoc);
3
alanine);
1
3
1
2,18 â pyrrole). C NMR (CDCl
alanine); 20.6(4C, CH CO); 20.8(4C, CH
Fmoc); 51.6(2C, CH alanine); 62.1(2C, C-6′ ose); 67.5(2C, CH
3
, 100 MHz) 18.3(2C, CH
3
3
3
3
3
CO); 47.2(2C, CH-9
2
68.4(3C, C-4′ ose); 71.4(3C, C-2′ or C-3′ ose); 72.3(3C, C-5′ose);
72.8(3C,C-2 or C-3′ose); 99.1(3C, C-1′ose); 112.9(1C, C meso);
115.1(6C, C-3,5 aryl); 119.3(2C, C-Fmoc); 119.7(2C, C meso);
120.2(1C, C meso); 121.2(1C, C-3 amidophenyl); 123.4(1C, C-5
amidophenyl); 124.0(1C, C-Fmoc); 124.1(1C, C-Fmoc); 126.3-
(1C, C-Fmoc); 126.5(1C, C-Fmoc); 127.0(1C, C-Fmoc); 127.1-
(1C, C-Fmoc); 129.6(1C, C-4 amidophenyl); 131.2(8C, C-â
pyrrole); 132.0(1C, C-1 amidophenyl); 134.6(1C, C-6 amido-
phenyl); 135.5(6C, C-2,6 aryl); 136.7(2C, C-1 aryl); 136.9(1C,
Fmoc); 68.5(2C, C-4′ ose); 71.4(2C, C-2′ or C-3′ ose); 72.3(2C,
C-5′ose); 72.9(2C,C-2 or C-3′ose); 99.2(2C, C-1′ose); 115.1(4C,
C-3,5 aryl); 118.2(4C, C-3,5 amidophenyl); 119.1(2C, C meso);
1
1
19.5(2C, C meso); 120.1(4C, C-Fmoc); 124.9(4C, C-Fmoc);
27.1(4C, C-Fmoc); 127.8(4C, C-Fmoc); 131.0(8C, C-â pyr-
role); 135.0(4C, C-2,6 amidophenyl); 135.5(4C, C-2,6 aryl);
1
37.2(2C, C-1 aryl); 137.5(2C, C-1 amidophenyl); 138.2(2C, C-4
IV
IV
amidophenyl); 141.3(4C, C Fmoc); 143.7(4C, C Fmoc); 146.2-
IV
(
4C, C-R Pyrrole); 156.7(4C, CO Fmoc and C-4 aryl); 169.4-
C-1 aryl); 138.1(1C, C-2 amidophenyl); 140.5(1C, C Fmoc);
140.6(1C, C Fmoc); 142.7(1C, C Fmoc); 142.9(1C, C Fmoc);
146.2(4C, C-R Pyrrole); 156.4(1C, CO Fmoc); 156.7(3C, C-4
IV
IV
IV
(2C, CH CO); 170.3(2C, CH CO); 170.6(4C, CH CO); 170.7(2C,
3
3
3
+
CO alanine). MS (MALDI) m/z: 1923.7 ([M + H] monoisotopic).

Glycosylated Amino Acid Porphyrin Derivatives
J . Org. Chem., Vol. 64, No. 12, 1999 4443
aryl); 170.1(1C, CO alanine); 169.4(6C, CH
CH CO); 170.5(3C, CH CO). MS (MALDI) m/z: 1961.6 ([M +
H] monoisotopic). Anal. Calcd for C104 O: C,
3.66; H, 5.24; N, 4.28. Found: C, 63.49; H, 5.20; N, 4.29.
-[4-(N-9-F lu or en ylm eth oxyca r bon yl)-L-a la n yla m in o)-
3
CO); 170.3(3C,
C-4 tolyl); 138.1(1C, C-2 amidophenyl); 139.2(3C, C-1 tolyl);
IV
IV
3
+
3
142.6(2C, C Fmoc); 142.8(2C, C Fmoc); 146.2(8C, C-R
pyrrole); 156.7(1C, CO Fmoc); 167.0(1C, CH CO); 169.1(1C,
CO); 170.1(1C, CO serine); 170.5(1C,
H
100
N
6
O
33‚H
2
3
6
CH
CH
3
3
CO); 169.5(1C, CH
CO). MS (MALDI) m/z: 1311.5 ([M + H] monoisotopic).
C, 72.38; H, 5.34; N, 6.41.
3
+
5
p h en yl]-10,15,20-t r is[4-(2′,3′,4′,6′-t et r a -O-a cet yl-â-D-glu -
cop yr a n osyloxy)p h en yl] P or p h yr in (17b). 12b (17 mg,
Anal. Calcd for C79
H
70
N
6
O
13
:
Found: C, 72.17; H, 5.31; N, 6.40.
0
.01 mmol), Fmoc-L-alanine (13 mg, 0.03 mmol), and dicyclo-
5-[4-(N-(9-F lu or en ylm et h oxyca r b on yl)-3-O-(2′,3′,4′,6′-
tetr a-O-acetyl-â-D-glu copyr an osyl)-L-ser ylam in o)ph en yl]-
10,15,20-tr istolylp or p h yr in (18b). 13b (7 mg, 0.01 mmol),
2 (20 mg, 0.03 mmol), and dicyclohexylcarbodiimide (6 mg, 0.03
hexylcarbodiimide (9 mg, 0.03 mmol) gave 18 mg of 17b (90%).
1
R
f
0.49 (toluene/acetone, 70/30). UV-visible (see Table 2). H
NMR (CDCl , 400 MHz) δ -2.87(2H, br s, NH); 2.03(6H, s,
OAc); 1.55(3H, d, J ) 6.8 Hz, CH alanine); 2.12(12H, s, OAc);
.13(6H, s, OAc); 2.23(12H, s, OAc); 3.98(3H, ddd, J ) 9.6-
.4-2.6 Hz, H-5′ose); 4.22(4H, m, H-9 Fmoc and H-6 ′ose);
.34(3H, dd, J ) 12.4-5.6 Hz, H-6 ′ose); 4.50(3H, m, CH
Fmoc); 5.23(3H, m, H-4′ose); 5.39(10H, m,
3
3
mmol) gave 11 mg of 18b (84%). R
methanol, 85/10/5). UV-visible (see Table 2). H NMR (400
MHz, CDCl ) δ -2.75(2H, br s, NH); 2.07(12H, s, OAc); 2.70-
(9H, s, Me tolyl); 3.88(1H, m, H-5′ ose); 4.0(1H, br t, J ) 8.2
Hz, H-â serine); 4.1(1H, dd, J ) 2.5-12.2 Hz, H-6 ′ ose); 4.18-
4.32(4H, m, H-6 ′ose, H-9 Fmoc, H-R and H-â serine); 4.53-
(3H, m, H-1′ ose and CH Fmoc); 5.10(1H, m, H-2′ ose);
f
0.54 (toluene/acetone/
1
2
5
4
b
3
a
alanine and CH
2
b
H-1′,2′,3′ ose and NH alanine); 7.24(2H, dt, J ) 7.5-1.2 Hz,
a
H-Fmoc); 7.28(6H, d, J ) 8.8 Hz, H-3,5 aryl); 7.33(2H, dt, J
2
)
7.5-1.1 Hz, H-Fmoc); 7.58(2H, dd, J ) 7.3-1.0 Hz,
5.18(1H, br t, J ) 9.3 Hz, H-4′ ose); 5.29(1H, br t, J ) 9.2 Hz,
H-3′ ose); 5.91(1H, br s, NH serine); 7.36(2H, br t, J ) 7.3 Hz,
H-Fmoc); 7.43(2H, br t, J ) 7.0 Hz, H-Fmoc); 7.55(6H, d, J
) 7.8 Hz, H-3,5 tolyl); 7.66(2H, br d, J ) 7.1 Hz, H-Fmoc);
7.79(2H, br d, J ) 7.1 Hz, H-Fmoc); 7.96(2H, d, J ) 7.9 Hz,
H-3,5 amidophenyl); 8.09(6H, d, J ) 7.8 Hz, H-2,6 tolyl); 8.20-
(2H, d, J ) 8.2 Hz, H-2,6 amidophenyl); 8.70(1H, br s, NH
amidophenyl); 8.86(8H, br s, H-â pyrrole). 13C NMR (100 MHz)
H-Fmoc); 7.69(2H, br d, J ) 7.5 Hz, H-Fmoc); 7.84(2H, d, J
)
8.6 Hz, H-3,5 amidophenyl); 8.06(6H, d, J ) 8.8 Hz, H-2,6
aryl); 8.08(2H, d, J ) 8.5 Hz, H-2,6 amidophenyl); 8.51(1H, br
s, NH-amidophenyl); 8.75(2H, d, J ) 4.9 Hz, H-3,7 â pyrrole);
8
.77(4H, s, H-12,13,17,18 â pyrrole); 8.79(2H, d, J ) 4.9 Hz,
1
3
H-2,8 â pyrrole). C NMR (CDCl
alanine); 20.6(12C, CH CO); 20.7(3C, CH
CO); 47.2(1C, CH-9 Fmoc); 51.4(1C, CH alanine); 62.1(3C, C-6′
ose); 67.6(1C, CH Fmoc); 68.5(3C, C-4′ ose); 71.4(3C, C-2′ or
C-3′ ose); 72.3(3C, C-5′ose); 72.9(3C,C-2 or C-3′ose); 99.2(3C,
C-1′ose); 115.1(6C, C-3,5 aryl); 118.0(2C, C-3,5 amidophenyl);
1
1
1
3
, 100 MHz) 18.2(1C, CH
CO); 20.8(3C, CH
3
3
3
3
-
20.6(2C, CH
(3C, Me tolyl); 47.2(1C, C-9 Fmoc); 54.7(1C, CH serine); 61.7-
(1C, CH Fmoc); 61.8(1C, C-6′ ose); 67.0(1C, CH serine);
3 3 3
CO); 20.7(1C, CH CO); 20.8(1C, CH CO); 21.5-
2
2
2
68.0(1C, C-4′ ose); 71.2(1C, C-2′ ose); 71.9(1C, C65′ ose); 72.5-
(1C, C-3′ ose); 102.2(C-1′ ose); 118.1(2C, C-3,5 amidophenyl);
119.0(1C, C meso); 120.1(2C, C Fmoc); 120.2(2C, C meso);
120.3(1C, C meso); 125.0(2C, C Fmoc); 127.2(2C, C Fmoc);
127.4(6C, C63,5 tolyl); 127.9(2C, C-Fmoc); 131.0(8C, C-â
pyrrole); 134.5(6C, C-2,6 tolyl); 135.1(2C, C-2,6 amidophenyl);
137.3(4C, C-4 tolyl and C-1 amidophenyl); 138.7(1C, C-4
amidophenyl); 139.2(3C, C-1 tolyl); 141.4(2C, CIV Fmoc); 143.6-
(2C, CIV Fmoc); 146.2(8C, CR-pyrrole); 156.7(1C, CO Fmoc);
19.3(3C, C meso); 119.5(1C, C meso); 120.1(2C, C-Fmoc);
24.9(2C, C-Fmoc); 127.2(2C, C-Fmoc); 127.8(2C, C-Fmoc);
31.0(8C, C-â pyrrole); 135.1(2C, C-2,6 amidophenyl); 135.5-
(
6C, C-2,6 aryl); 137.2(3C, C-1 aryl); 137.5(1C, C-1 amido-
IV
phenyl); 138.2(1C, C-4 amidophenyl); 141.4(2C, C Fmoc);
IV
1
43.8(2C, C Fmoc); 146.5(4C, C-R Pyrrole); 156.7(4C, C-4 aryl
and CO Fmoc); 169.4(6C, CH CO); 170.3(4C, CH CO and CO
alanine); 170.5(3C, CH CO). MS (MALDI) m/z: 1961.6 ([M +
H] monoisotopic). Anal. Calcd for C104 O: C,
1.96; H, 5.30; N, 4.16. Found: C, 62.09; H, 5.33; N, 4.19.
-[2-(N-(9-F lu or en ylm et h oxyca r b on yl)-3-O-(2′,3′,4′,6′-
3
3
3
+
H
100
N
6
O
33‚3H
2
169.5(2C, CH
3 3 3
CO); 169.6(1C, CH CO); 170.2(1C, CH CO);
6
170.7(1C, CO serine). MS (MALDI) m/z: 1311.6 ([M +
+
5
70 6
H] monoisotopic). Anal. Calcd for C79H N O13: C, 72.38; H,
tetr a-O-acetyl-â-D-glu copyr an osyl)-L-ser ylam in o)ph en yl]-
5.34; N, 6.41. Found: C, 72.21; H, 5.31; N, 6.42.
Gen er a l P r oced u r e for Syn th esis of Com p ou n d s 19a ,b,
20, 21, a n d 22a ,b. A 20 mg portion of the porphyrin was
dissolved in 2 mL of CH Cl /morpholine (1/1), and the mixture
2 2
was stirred for 60 min at room temperature. After evaporation
under vacuum, the crude product was dissolved in 2 mL of
1
2
0
0,15,20-tr istolylp or p h yr in (18a ). 13a (20 mg, 0.03 mmol),
(59 mg, 0.09 mmol), and dicyclohexylcarbodiimide (19 mg,
.09 mmol) gave 23 mg of 18a (60%). R
f
0.70 (CH
2
Cl
2
). UV-
1
visible (see Table 2). H NMR (400 MHz, CDCl
3
) δ 2.75(2H,
br s, NH); -1.11(3H, s, OAc); 1.24(3H, s, OAc); 1.84(3H, s,
OAc); 2.00(3H, s, OAc); 2.70(9H, s, Me tolyl); 3.20-3.38(3H,
2 2
CH Cl /MeOH (80/20). Sodium methanolate in dry methanol
m, H-9 Fmoc, H-R and H-â serine); 3.52(2H, m, CH
2
Fmoc);
(1.5 equiv/OAc, 1 M) was added, and the mixture was stirred
for 60 min at room temperature. Then porphyrin was precipi-
tated by addition of petroleum ether.
5-[2-(â-D-glu cop yr a n osyloxy)p h en yl]-10,15,20-tr is[4-(L-
a la n yla m in o)p h en yl]p or p h yr in (19a ). The title compound
was purified by gel filtration on a Sephadex LH20 column
3
.65(1H, ddd, J ) 2.5-5.1-9.7 Hz, H-5′ ose); 3.78(1H, m, H-â
b
serine); 4.00(1H, dd, J ) 2.6-12.1 Hz, H-6 ′ose); 4.12(1H, dd,
J ) 5.2-12.1 Hz, H-6
a
′ ose); 4.18(1H, dd, J ) 2.2-7.8 Hz, H-2′
ose); 4.30(1H, m, NH serine); 4.64(1H, t, J ) 9.2 Hz, H3′ ose);
.70(1H, t, J ) 9.2 Hz, H-4′ ose); 4.91(1H, d, J ) 7.8 Hz, H-1′
4
ose); 6.20(2H, dt, J ) 1.2-7.6 Hz, H Fmoc); 6.31(2H, m, H
Fmoc); 6.40(2H, br d, J ) 7.4 Hz, H Fmoc); 6.70(2H, m, H
Fmoc); 7.54(1H, m, H-4 amidophenyl); 7.56(6H, d, J ) 7.9 Hz,
H-3,5 tolyl); 7.85(1h, dt, J ) 1.3-7.9 Hz, H-3 amidophenyl);
eluted with THF/H
2 f 2 2
O (8/2), (8 mg, 71%). R 0.45 (CH Cl /
MeOH, 70/30). UV-visible (see Table 2). MS (MALDI) m/z:
+
1051.5 ([M + H] monoisotopic). Anal. Calcd for C59
58 10 9
H N O :
C, 67.44; H, 5.52; N, 13.33. Found: C, 67.36; H, 5.50, N, 13.30.
5-[4-(â-D-glu cop yr a n osyloxy)p h en yl]-10,15,20-tr is[4-(L-
a la n yla m in o)p h en yl]p or p h yr in (19b). The title compound
was purified by gel filtration on a Sephadex LH20 column
8
7
8
.03(1H, dd, J ) 1.2-7.6 Hz, H-5 amidophenyl); 8.10(6H, J )
.9 Hz, H-2,6 tolyl); 8.68(1H, d, J ) 8.1 Hz, H-2 amidophenyl);
.86(1H, br s, H-6 amidophenyl); 8.75(1H, d, J ) 4.9 Hz, H-3
or H-7 â pyrrole); 8.77(1H, d, J ) 4.9 Hz, H-3 or H-7 â pyrrole);
8
4
eluted with THF/H
2 f 2 2
O (8/2), (8 mg, 71%). R 0.43 (CH Cl /
.82(1H, d, J ) 4.9 Hz, H-2 or H-8 â pyrrole); 8.86(1H, d, J )
.9 Hz, H-2 or H-8 â pyrrole); 8.91(4H, s, H-12,13,17,18 â
MeOH, 70/30). UV-visible (see Table 2). MS (MALDI) m/z:
+
1051.5 ([M+H) monoisotopic). Anal. Calcd for C59
58 10 9
H N O :
1
3
pyrrole). C NMR (100 MHz) 17.3(1C, CH
CO); 20.4(1C, CH CO); 20.7(1C, CH CO); 21.5(3C, Me tolyl);
5.7(1C, C-9 Fmoc); 51.3(1C, CH serine); 61.3(1C, C-6′ ose);
5.5(1C, CH Fmoc); 66.5(1C, CH serine); 67.8(1C, C-4′ ose);
0.1(C-2′ ose); 71.2(C-5′ ose); 72.1(1C, C-3′ ose); 102.4(1C,
3
CO); 19.8(1C, CH
3
-
C, 67.44; H, 5.52; N, 13.33. Found: C, 67.39, H, 5.53, N, 13.29.
5,15-Bis[4-(L-a la n yla m in o)p h en yl]-10,20-bis[4-(â-D-glu -
cop yr a n osyloxy)p h en yl]p or p h yr in (20). The title com-
pound was purified by gel filtration on a Sephadex LH20
3
3
4
6
7
2
2
column eluted with THF/H
Cl /EtOH/H O, 4/6/2). UV-visible (see Table 2). MS (MALDI)
m/z: 1143.6 ([M
14: C, 65.17; H, 5.42; N, 9.80. Found: C, 65.02; H,
2 f 2
O (9/1), (9 mg, 75%). R 0.57 (CH -
C-1′ose); 114.1(1C, C meso); 119.5(2C, C-Fmoc); 120.5(2C, C
meso); 121.0(1C, C meso); 121.3(1C, C-3 amidophenyl); 12.3.4-
2
2
+
+
H] monoisotopic). Anal. Calcd for
(
1
1C, C-5 amidopheyl); 124.0(2C, C-Fmoc); 126.6(2C, C-Fmoc);
27.1(2C, C-Fmoc); 127.4(6C, C-3,5 tolyl); 129.6(1C, c-4 ami-
dophenyl); 130.9(8C, C-â pyrrole); 132.0(1C, C-1 amidophenyl);
34.4(6C, C-2,6 tolyl); 134.7(1C, C-6 amidophenyl); 137.3(3C,
62 62 8
C H N O
5.40; N, 9.81.
5,10-Bis[4-(L-a la n yla m in o)p h en yl]-15,20-bis[4-(â-D-glu -
cop yr a n osyloxy)p h en yl]p or p h yr in (21). The title com-
1

4
444 J . Org. Chem., Vol. 64, No. 12, 1999
Sol et al.
pound was purified by gel filtration on a Sephadex LH20
column eluted with THF/H O (9/1), (8 mg, 67%). R 0.54 (CH
Cl /EtOH/H O, 4/6/2). UV-visible (see Table 2). MS (MALDI)
m/z: 1143.6 ([M + H] monoisotopic). Anal. Calcd for
14: C, 65.17; H, 5.42; N, 9.80. Found: C, 65.07; H,
.39; N, 9.78.
18a ,b was dissolved in 2 mL of CH
mixture was stirred for 60 min at room temperature. After
evaporation under vacuum, the acetate groups of the carbo-
hydrate moieties were removed according to the literature.
The crude product was dissolved in 2 mL of hydrazine/MeOH/
2
Cl
2
/morpholine (1/1). The
2
f
2
-
2
2
+
18
62 62 8
C H N O
5
2 2
CH Cl (4/3/3), and the mixture was stirred for 2 h. Afterward,
5
-[2-(L-Ala n yla m in o)p h en yl]-10,15,20-t r is[4-(â-D-glu -
acetone (2 mL) was added to the solution to remove excess
hydrazine. The porphyrin was precipitated by addition of
petroleum ether, filtered, and then purified by gel filtration
cop yr a n osyloxy)p h en yl]p or p h yr in (22a ). The title com-
pound was purified by gel filtration on a Sephadex LH20
column eluted with MeOH/H
Cl /EtOH/H O, 4/6/2). UV-visible (see Table 2). MS (MALDI)
m/z: 1235.5 ([M + H] monoisotopic). Anal. Calcd for
19: C, 63.23; H, 5.35; N, 6.81. Found: C, 63.18; H,
.31; N, 6.78.
2 f 2
O (9/1), (9 mg, 71%). R 0.52 (CH -
on a Sephadex LH20 column, eluted with THF/H
5-[2-(3-O-(â-D-glu cop yr a n osyl)-L-ser yla m in o)p h en yl]-
0,15,20-tr istolylp or p h yr in (23a ): 10 mg (71%). R 0.48
CH Cl /MeOH,80/20). UV-visible (see Table 2). MS (MALDI)
m/z: 920.4 ([M
: C, 73.14; H, 5.55; N, 9.14. Found: C, 73.07; H,
2
O (9/1).
2
2
+
1
f
65 66 6
C H N O
(
2
2
5
+
+
H] monoisotopic). Anal. Calcd for
5
-[4-(L-Ala n yla m in o)p h en yl]-10,15,20-t r is[4-(â-D-glu -
cop yr a n osyloxy)p h en yl]p or p h yr in (22b). The title com-
56 51 6 7
C H N O
pound was purified by gel filtration on a Sephadex LH20
5.56; N, 9.10.
column eluted with MeOH/H
Cl /EtOH/H O, 4/6/2). UV-visible (see Table 2). MS (MALDI)
m/z: 1235.7 ([M + H] monoisotopic). Anal. Calcd for
19: C, 63.23; H, 5.35; N, 6.81. Found: C, 63.14; H,
2
O (9/1), (8 mg, 64%). R
f
049 (CH
2
-
5-[4-(3-O-(â-D-glu cop yr a n osyl)-L-ser yla m in o)p h en yl]-
2
2
1
0,15,20-tr istolylp or p h yr in (23b): 8 mg (57%). R
Cl /MeOH,80/20). UV-visible (see Table 2). MS (MALDI) m/z:
20.4 ([M + H] monoisotopic). Anal. Calcd for C56
f 2
0.42 (CH -
+
2
C
65
H
66
N
6
O
+
9
51 6 7
H N O :
5
.37; N, 6.81.
C, 73.14; H, 5.55; N, 9.14. Found: C, 73.04; H, 5.53; N, 9.15.
P r oced u r e for Dep r otection of Mon oglu cosylser yl-
tr istolylp or p h yr in s 23a ,b. A 20 mg portion of porphyrin
J O982499+