pheophytin Jukart..
서론
재료및방법
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650 nm (10).
3. 124 15 3 68 2 3 124 2, 3, 43 5, 5 23 (5). 4. 5 4 5 53. 650 nm (10),..,.
16 pheophytin Jukart.. IC 50 1mg/ml..
pheophytin Jukart.. IC 50 1mg/ml.
(a)
CH 3 CH 3 H H H H O O COOMe O O O HO COOMe O Pheophytin a 10-Hydroxy pheophytin a
000921 silk06 : Prediction of degradation 430 nm 80000 70000 60000 50000 40000 30000 20000 10000 0-10000 0 20 40 60 80 100 Retention Time, minutes A
1 2 3 4 5 6 700 350 Area 600 500 400 300 200 100 0 0.25ul 0.5ul 1ul 2ul 4ul Dose of CpD Fluorescence Area 1 of CpD Treated Cells Tumor ormal Area 300 250 200 150 100 50 0 0.25ul 0.5ul 1ul 2ul 4ul Dose of CpD Fluorescence Area 2 of CpD Treated Cells Tumor ormal M ultiplicity (Tum or/orm al) 15 10 5 0 0.25ul 0.5ul 1ul 2ul 4ul Dose of CpD A1 T/ Selective Intake of CpD by Tumor Cells A2 T/
60 Apoptotic response (%) 50 40 30 20 10 0 0 60 120 180 240 300 CpD dose (ug) Jurkat HL-60 HeLa MCF7 MCF7/ADR HepG2 Fig. 6. The CpD dose dependence of the apoptotic fraction measured by flow cytometery among human tumnor cell lines. F1 F2 F3 F4
pheophorbide a H H HOOC H H 3 COOC O Molecular Weight =592.70 Exact Mass =592 Molecular Formula =C35H364O5 Molecular Composition =C 70.93% H 6.12% 9.45% O 13.50%
O OH OH O OH OH O anthrarufin (1,5-DHAQ) 1,8-dihydroxyanthraquinon e (1,8-DHAQ) O (a) (b) Intensity (a.u.) Intensity (a.u.) -2 0 2 4 6 8-2 0 2 4 6 8 Time (µs) -10 0 10 20 30 40-10 0 10 20 30 40 Time (µs)
H 3 CO OCH 3 S S S S S S H 3 CO OCH 3 meso-tetraphenyl-trithiasapphyrin meso-tetra(p-methoxyphenyl)-trithiasapphyrin ((p-meo)-s 3 TPS) (S 3 TPS)
H 2 TPP OTPPH STPPH OSTPP H H O H S H O S 12.
Absorption (arb. units) (a) H 2TPP OTPPH STPPH 10 400 500 600 700 W avelength (nm ) Absorption (arb. units) (b) 500 600 700 400 500 600 700 W avelength (nm ) Fluorescence (arb. units) (c) 600 650 700 750 800 W avelength (nm ) H 2 TPP OTPPH STPPH OSTPP
Intensity (arb. uints) IRF Partially degassed Air-saturated 0 5 10 15 Time (ns) 0.10 0.04 O.D. 0.05 O.D. 0.02 0.00 0 25 50 75 100 Excitation Power (% ) 0.00 0 5 10 15 20 Time (µs)
Intensity (arb. units) OTPPH OSTPP 0 25 50 75 Time (µs)
R R R R H H R R R R Ge (a) 2 x 20 H 2 TPP (p-oh) 4 TPPH 2 (b) GeTPPCl 2 Ge(p-OH) 4 TPPCl 2 O. D. 1 Intensity (a.u.) 0 300 400 500 600 700 Wavelength (nm) 600 700 800 W avelength (nm) 30000 IRF (a) Counts 20000 10000 (b) (c) (d) 0 0 5 10 15 Time (ns)
0.06 S S O. D. 0.04 0.02 x 5 S S 0.00 200 400 600 800 1000 1200 W avelength (nm ) 21.
(a) (b) Intensity (a.u.) 1 2 1-10 0-10 0 10 20 30 40 50 10 20 30 40 50 Time (µs) Time (µs) Intensity (a.u.)
Rhodamine B Two Photon Raw Spectrum Rhodamine B Power Dependence Intensity (a.u.) 750 800 850 900 950 1000 1050 Wavelength (nm) Log ( Intensity ) 10 20 30 40 50 60 70 80 Power ( kw )
8 6 8 6 log pfu 4 log pfu 4 2 2 0 Untreated Light Acetone 0 0 1.8753.75 7.5 15 30 60 CpD dose (µg/ml)
3H-uridine incorpartion (cpm) 14000 12000 10000 8000 6000 4000 2000 0 0 3.75 7.5 30 CpD dose (µg/ml)
CpD dose(µg/ml) 8000 3 H-UTP incorporation (cpm) 6000 4000 2000 0 0 3.75 7.5 30 /C
F
(B)
pheophorbide a pyropheophorbide a H H H H HOOC H H 3 COOC O HOOC H H O Molecular Weight =592.70 Exact Mass =592 Molecular Formula =C35H364O5 Molecular Composition =C 70.93% H 6.12% 9.45% O 13.50% Molecular Weight =534.66 Exact Mass =534 Molecular Formula =C33H344O3 Molecular Composition =C 74.13% H 6.41% 10.48% O 8.98% R 3 R 3 R 2 R 1 R 1 R 2 R 1 R 1 R 2 H R 3 R 3 H H R 2 R 1 R 1 R 2 PF : R1, R2, R3 = F PC : R1, R2 = H, R3 = CO2CH3 PO : R1, R2, R3 = H [5, 10, 15-tris(pentafluorophenyl)corrole] [5, 10, 15-triphenylcorrole] [5, 10, 15-tris(p-methoxycarbonylphenyl)corrole]
O.D. (normalized) 1.0 0.5 0.2 0.1 0.0 500 600 700 0.0 400 500 600 700 W avelength (nm )
Intensity (normalized) 1.0 0.5 0.0 600 700 800 W avelength (nm )
Substance Φ " f λ f /nm ε PF 0.12 646 17078 Intensity (arb. units) PO PC H 2 TPP 0.14 0.16 0.09 * 666 14892 675 15784 16907-10 0 10 20 30 40 Time (ns)
0.09 0.04 O. D. 0.06 0.03 O. D. 0.03 0.02 0.01 0.00 0 20 40 60 80 100 Transmittance (%) 0.00-50 0 50 100 150 200 Time (µs) F Table 4. Triplet state quantum yields
Substance Φ T PF 0.85 PC 0.72 PO 0.67 H 2 TPP 0.72*
. ( ).. 28s.. 25-29s. 28-29s.........
Intensity (a.u.) 0.004 0.002 0.000 (a) Intensity (a.u.) 0.004 0.002 0.000 (b) Intensity (a.u.) 0.004 0.002 0.000-20 0 20 40 60 80 100 Time (µs) (c)
(a) (a) (b) (b) Current (a.u.) Current (a. u.) Current (a. u.) (c) 2.0 1.5 1.0 0.5 Potential (V vs A g/a gc l) 0.0 (c)
Substance PF PC PO E pa 0.92 0.83 0.71 H 2 TPP 1.07
Substance PF PC PO Φ f 0.14 0.18 0.16 k f / 10 6 s -1 / 10 6 s -1 / 10 6 s -1 / 10 6 s -1 k r k ic k isc 167 24 1 142 175 32 17 126 172 27 26 119 Φ T 0.85 0.72 0.67 PF PC PO Φ T 0.87 0.76 0.72 τ p 2 Φ S T T / µs O 0.82 24 0.94 ~1 0.62 16 0.82 ~1 0.49 14 0.68 ~1 E pa 0.92 0.83 0.71 q k /10 9 s -1 0.24 0.95 1.61 Substance k b f Φ f k f k T k T Φ / 10 6 s -1 / 10 6 s -1 / 1 0 3 s -1 / 10 6 s -1 T PF 167 189 0.12 17.7 3.9 0.85 PC 175 208 0.16 23.1 6.0 0.72 PO 172 200 0.14 33.7 6.3 0.67
2~3 4~6.
H H H H H H CO 2 Me O CO 2 Me O O Phytyl O O
(a) Voyager Spec #1=>BC[BP = 535.8, 63173] 100 90 535.8139 593.8971 534.7965 6.3E+4 80 70 536.8103 592.8835 594.8999 % Intensity 60 50 40 610.9296 30 20 10 537.8135 608.9074 533.7900 625.9513 549.8071 595.9040 538.8122 611.9366 437.6604 481.7225 540.8050 627.9699 445.4667 519.7630 581.9254 632.9480 758.2881 0 0 423.0 547.2 671.4 795.6 919.8 1044.0 Mass (m/z)
(b) 100 90 Voyager Spec #1=>BC[BP = 535.8, 63173] 535.8139 593.8971 534.7965 6.3E+4 80 592.8835 609.9237 536.8103 70 594.8999 % Intensity 60 50 40 610.9296 30 608.9074 537.8135 533.7900 625.9513 20 595.9040 607.9052 549.8071 536.2860 594.4060 611.9366 626.9554 536.0727 10 551.8197 610.4446 525.7987 565.8258 593.3955 631.9355 544.6924 581.9254 592.6539 612.9530 554.1771 643.9991 629.9648 0 0 510.0 541.2 572.4 603.6 634.8 666.0 Mass (m/z) H 3 C H H H HOOC H O C 33 H 34 4 O 3 Exact Mass: 534.26 Mol. Wt.: 534.65 C, 74.13; H, 6.41;, 10.48; O, 8.98 H 3 C H H H O COOH CO 2 Me C 35 H 36 4 O 5 Exact Mass: 592.27 Mol. Wt.: 592.68 C, 70.93; H, 6.12;, 9.45; O, 13.50 H 3 C O O H H H O CO 2 Me C 55 H 74 4 O 5 Exact Mass: 870.57 Mol. Wt.: 871.20 C, 75.83; H, 8.56;, 6.43; O, 9.18 H H
Voyager Spec #1=>BC[BP = 284.6, 54548] 100 284.6522 5.5E+4 90 80 535.8530 70 % Intensity 60 50 40 30 20 172.2474 413.7105 10 234.2871 593.9298 429.6674 274.5859 0 0 99.0 479.2 859.4 1239.6 1619.8 2000.0 Mass (m/z)
Voyager Spec #1=>BC[BP = 284.7, 49772] 100 284.6779 5.0E+4 90 80 70 % Intensity 60 50 535.9012 40 30 172.2542 190.2923 20 10 379.5359 212.2937 549.8999 205.4219 429.7239 566.0001 872.5610 888.5559 0 0 99.0 479.2 859.4 1239.6 1619.8 2000.0 Mass (m/z) O H H HO MeO 2 C O O H 3 C O H H HO MeO 2 C O O H 3 C MeO 2 C H H MeO 2 C O
O.D.(normalized) 1 0 O.D.(normalized) 0.09 0.06 0.03 0.00 460 480 500 520 540 560 Wavelength(nm) 400 500 600 700 Wavelength(nm) Fluorescence Intensity(normalized) 1 0 600 650 700 750 800 Wavelength(nm) f f /nm(max)
f / ns f / ns (degassed) (air saturated) k s q / 10 10 s -1 0.04 (a) O.D. 0.02 0.00-100 0 100 200 300 400 500 0.06 Time( µs) O.D. 0.04 0.02 (b) 0.00 0.0 0.5 1.0 1.5 Time( µs) (a) degassed condition (b) air saturated condition
T / s (degassed) T / s (air saturated) /s
1265 1231 TiO2-HP HP 1288 1247 4000 3000 2000 1000 wave number(cm -1 )
CH 3 CH 3 H H H H O O COOMe O O O HO COOMe O Pheophytin a 10-Hydroxy pheophytin a
X CH 3 CH 3 CH 3 H H H H H H O OR COOMe O O OR HO COOMe O O OR HO COOMe O Dried, powdered Silkworm W aste (200 g) / water Filter Solid part was collected, dried Extracted with MeOH/chloroform Filter Remove solvent The mixture of chlorin derivatives (4-7 g) Column chromatography Pheophytin a (0.7 ~1.0 g, pure compound) Yellow pigments Other minor parts 10-hydroxypheophytin a (~100 mg, pure) Polar porphyrins
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