Absorption, Distribution and Excretion
Particle size should be optimized to achieve targeted and extended drug delivery to the affected tissues. We describe here the effects of the mean particle size on the pharmacokinetics and photothrombic activity of meso-tetra(carboxyphenyl)porphyrin (TCPP), which is encapsulated into biodegradable nanoparticles based on poly(d,l-lactic acid). Four batches of nanoparticles with different mean sizes ranging from 121 to 343 nm, were prepared using the emulsification-diffusion technique. The extravasations of each TCPP-loaded nanoparticle formulation from blood vessels were measured, as well as the extent of photochemically induced vascular occlusion. These preclinical tests were carried out in the chorioallantoic membrane (CAM) of the chicken's embryo. Fluorescence microscopy showed that both the effective leakage of TCPP from the CAM blood vessels and its photothrombic efficiency were dependent on the size of the nanoparticle drug carrier. Indeed, the TCPP fluorescence contrast between the blood vessels and the surrounding tissue increased at the applied conditions, when the particle size decreased. This suggests that large nanoparticles are more rapidly eliminated from the bloodstream. In addition, after injection of a drug dose of 1 mg/kg body weight and a drug-light application interval of 1 min, irradiation with a fluence of 10 J/sq cm showed that the extent of vascular damage gradually decreased when the particle size increased. The highest photothrombic efficiency was observed when using the TCPP-loaded nanoparticles batch with a mean diameter of 121 nm. Thus, in this range of applied conditions, for the treatment of for instance a disease like choroidal neovascularization (CNV) associated with age-related macular degeneration (AMD), these experiments suggest that the smallest nanoparticles may be considered as the optimal formulation since they exhibited the greatest extent of vascular thrombosis as well as the lowest extravasation. /TCPP nanoparticles/
We studied the uptake of meso-tetra (carboxyphenyl) porphyrin (TCPP) nanoparticles by SW480 cells and carried out a systematic investigation of the cellular internalization mechanism of TCPP nanoparticles, also studied the photocytotoxicity of TCPP nanoparticles. At first, meso-tetra (carboxyphenyl) porphyrin (TCPP) nanoparticles were prepared by the method of mixing solvent techniques. SW480 cellular uptakes of photosensitizers (TCPP nanoparticles, TCPP-loaded poly (lactic-co-glycolic acid) (PLGA) nanoparticles and free TCPP) were analyzed by the method of fluorospectrophotometry. Endocytosis mechanism investigation was carried out by preincubating SW480 cells at 4 degrees C, and preincubating SW480 cells with sucrose, K+-free buffer solution and filipin. Clathrin HC expression after incubating SW480 cells with these three photosensitizers was analyzed by methods of Western blot and RT-PCR. At last, we analyzed the photo-cytotoxicity after incubating SW480 cells with photosensitizers and receiving irradiation. SW480 cells showed rapid uptake (0.0083 fmoles TCPP/cell) of TCPP nanoparticles after 1h incubation. We also demonstrated that the uptake of TCPP nanoparticles by SW480 cells was a clathrin-mediated endocytosis pathway. As a result of rapid internalization of TCPP nanoparticles by SW480 cells, this special photosensitizer showed very high photocytotoxic effect on SW480 cells in vitro. The nano-sized photosensitizer with no matrix cover: TCPP nanoparticles, can produce higher photocytotoxicity than other photosensitizers (TCPP-loaded PLGA nanoparticles and free TCPP). The in vivo tumor growth inhibition experiment indicated that TCPP nanoparticles plus PDT treatment induced the most dramatic tumor-inhibiting efficacy in all TCPP treated groups. The results of this study suggest that TCPP nanoparticles represent a potential and powerful photodynamic therapy agent. /TCPP nanoparticles/
Athymic nude mice with subcutaneously xenotransplanted urothelial carcinoma received intravenously injections of the synthetic sensitizer meso-tetra (4-carboxyphenyl) porphyrin. Fluorescence at the tumors and the skin was excited using a Kr+ laser (407 nm) and was detected at 652 nm using a fiberoptical sensor in combination with an optical multichannel analyzer at different times after application of the sensitizer. Photodynamic treatment was carried out using an Ar+ laser pumped dye laser (650 nm; 100 mW/sq cm; 100 J/sq cm). The delay of tumor growth was determined in a follow-up period of four weeks and was correlated to the in vivo fluorescence measurements.
Mesotetra(4-carboxyphenyl)porphyrin (mTCPP) is a commercially available small molecule fluorophore and photosensitizer with four free carboxylic acid groups. mTCPP can readily be conjugated with amines for facile attachment of functional groups. In this work, we synthesized and assessed tetravalent, lysine-conjugated mTCPP, for its potential applications in targeted imaging and photodynamic therapy. Fmoc-protected d-lysine or l-lysine was conjugated to mTCPP via amide coupling with the epsilon amine group of lysine, followed by Fmoc deprotection. The resulting compounds did not dissolve well in aqueous solvent, but could be solubilized with the assistance of surfactants, including cholic acid. The l-amino acid transporter (LAT1) can uptake diverse neutral l-amino acids. In vitro studies with U87 cells revealed a non-specific uptake of the hydrophobic Fmoc-protected lysine-conjugated mTCPP precursors, but not d- or l-lysine mTCPP. Likewise, only the Fmoc-protected compounds induced substantial phototoxicty in cells following incubation and irradiation with blue light. These experimental results do not provide evidence to suggest that lysine-mTCPP is able to specifically target cancer cells. However, they do highlight mTCPP as a convenient and accessible framework for assessing molecular targeting of photosensitizers. /lysine-conjugated mTCPP/

Toxicity Summary
IDENTIFICATION AND USE: Tetracarboxyphenylporphine (TCPP) is a dark blue to purple to black powder. TCPP is a potent photosensitizer activated by visible light. It has been used as a research chemical, chemical intermediate, and porphyrin dye. It has been tested for photodynamic therapy of cancer in animal cell models, and as a diagnostic noninvasive assay in lung cancer patients. HUMAN STUDIES: There are no data available. ANIMAL STUDIES: The photosensitizing effects of TCPP on ColE1 supercoiled DNA were studied using agarose gel electrophoresis. Photoinduced single- and double-strand breaks were observed to form under neutral conditions.

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Interactions
The role of the neoplastic cell in both porphyrin localization and the photochemotherapeutic response was investigated with the use of a series of tumor-localizing porphyrins and the L1210 tumor system. In vivo photoirradiation of DBA/2Ha mice bearing L1210 solid tumors and previously given injections of meso-tetra-(4-sulfonatophenyl)-porphine, meso-tetra-(4-carboxyphenyl)-porphine, or hematoporphyrin derivative (Hpd) indicated that all three chemicals elicited a photodynamic response resulting in necrosis of exposed tissue. Isolation of tumor cells from mice given injections of porphyrin with the use of mild mechanical means and physiologic conditions followed by in vitro photoirradiation of the cells under conditions established to optimize rapid cytocidal effects resulted in no appreciable cell death. A similar situation was noted with the use of spleen cells from mice given injections of Hpd, the spleen cells presumably containing substantial amounts of porphyrin. Both fluorescence microscopy and chemical extraction and quantitation of the porphyrins in the cells indicated that the inability to elicit a rapid cytocidal effect upon in vitro photoirradiation resulted from either the absence of or the presence of only very small amounts of porphyrin. These results indicate that in this particular tumor system the neoplastic cell per se plays only a minor role in porphyrin localization and, as a consequence, cannot be readily killed upon photoirradiation, suggesting that rapid cytocidal effects, due solely to porphyrin contained within the cell, probably do not occur among the majority of parenchymal cells during in vivo photoirradiation.

[1]. Removal of Cu(II) from water by tetrakis(4-carboxyphenyl) porphyrin-functionalized mesoporous silica. J Hazard Mater. 2011 Jan 30;185(2-3):1311-7.

Therapeutic Uses
/EXPL THER/ Particle size should be optimized to achieve targeted and extended drug delivery to the affected tissues. We describe here the effects of the mean particle size on the pharmacokinetics and photothrombic activity of meso-tetra(carboxyphenyl)porphyrin (TCPP), which is encapsulated into biodegradable nanoparticles based on poly(d,l-lactic acid). Four batches of nanoparticles with different mean sizes ranging from 121 to 343 nm, were prepared using the emulsification-diffusion technique. The extravasations of each TCPP-loaded nanoparticle formulation from blood vessels were measured, as well as the extent of photochemically induced vascular occlusion. These preclinical tests were carried out in the chorioallantoic membrane (CAM) of the chicken's embryo. Fluorescence microscopy showed that both the effective leakage of TCPP from the CAM blood vessels and its photothrombic efficiency were dependent on the size of the nanoparticle drug carrier. Indeed, the TCPP fluorescence contrast between the blood vessels and the surrounding tissue increased at the applied conditions, when the particle size decreased. This suggests that large nanoparticles are more rapidly eliminated from the bloodstream. In addition, after injection of a drug dose of 1 mg/kg body weight and a drug-light application interval of 1 min, irradiation with a fluence of 10 J/sq cm showed that the extent of vascular damage gradually decreased when the particle size increased. The highest photothrombic efficiency was observed when using the TCPP-loaded nanoparticles batch with a mean diameter of 121 nm. Thus, in this range of applied conditions, for the treatment of for instance a disease like choroidal neovascularization (CNV) associated with age-related macular degeneration (AMD), these experiments suggest that the smallest nanoparticles may be considered as the optimal formulation since they exhibited the greatest extent of vascular thrombosis as well as the lowest extravasation. /TCPP nanoparticles/
/EXPL THER/ The role of the neoplastic cell in both porphyrin localization and the photochemotherapeutic response was investigated with the use of a series of tumor-localizing porphyrins and the L1210 tumor system. In vivo photoirradiation of DBA/2Ha mice bearing L1210 solid tumors and previously given injections of meso-tetra-(4-sulfonatophenyl)-porphine, meso-tetra-(4-carboxyphenyl)-porphine, or hematoporphyrin derivative (Hpd) indicated that all three chemicals elicited a photodynamic response resulting in necrosis of exposed tissue. Isolation of tumor cells from mice given injections of porphyrin with the use of mild mechanical means and physiologic conditions followed by in vitro photoirradiation of the cells under conditions established to optimize rapid cytocidal effects resulted in no appreciable cell death. A similar situation was noted with the use of spleen cells from mice given injections of Hpd, the spleen cells presumably containing substantial amounts of porphyrin. Both fluorescence microscopy and chemical extraction and quantitation of the porphyrins in the cells indicated that the inability to elicit a rapid cytocidal effect upon in vitro photoirradiation resulted from either the absence of or the presence of only very small amounts of porphyrin. These results indicate that in this particular tumor system the neoplastic cell per se plays only a minor role in porphyrin localization and, as a consequence, cannot be readily killed upon photoirradiation, suggesting that rapid cytocidal effects, due solely to porphyrin contained within the cell, probably do not occur among the majority of parenchymal cells during in vivo photoirradiation.
/EXPL THER/ We studied the uptake of meso-tetra (carboxyphenyl) porphyrin (TCPP) nanoparticles by SW480 cells and carried out a systematic investigation of the cellular internalization mechanism of TCPP nanoparticles, also studied the photocytotoxicity of TCPP nanoparticles. At first, meso-tetra (carboxyphenyl) porphyrin (TCPP) nanoparticles were prepared by the method of mixing solvent techniques. SW480 cellular uptakes of photosensitizers (TCPP nanoparticles, TCPP-loaded poly (lactic-co-glycolic acid) (PLGA) nanoparticles and free TCPP) were analyzed by the method of fluorospectrophotometry. Endocytosis mechanism investigation was carried out by preincubating SW480 cells at 4 degrees C, and preincubating SW480 cells with sucrose, K+-free buffer solution and filipin. Clathrin HC expression after incubating SW480 cells with these three photosensitizers was analyzed by methods of Western blot and RT-PCR. At last, we analyzed the photo-cytotoxicity after incubating SW480 cells with photosensitizers and receiving irradiation. SW480 cells showed rapid uptake (0.0083 fmoles TCPP/cell) of TCPP nanoparticles after 1h incubation. We also demonstrated that the uptake of TCPP nanoparticles by SW480 cells was a clathrin-mediated endocytosis pathway. As a result of rapid internalization of TCPP nanoparticles by SW480 cells, this special photosensitizer showed very high photocytotoxic effect on SW480 cells in vitro. The nano-sized photosensitizer with no matrix cover: TCPP nanoparticles, can produce higher photocytotoxicity than other photosensitizers (TCPP-loaded PLGA nanoparticles and free TCPP). The in vivo tumor growth inhibition experiment indicated that TCPP nanoparticles plus PDT treatment induced the most dramatic tumor-inhibiting efficacy in all TCPP treated groups. The results of this study suggest that TCPP nanoparticles represent a potential and powerful photodynamic therapy agent. /TCPP nanoparticles/
/EXPL THER/ Introduction: Early detection of lung cancer in high-risk individuals reduces mortality. Low-dose spiral computed tomography (LDCT) is the current standard but suffers from an exceedingly high false-positive rate (>96%) leading to unnecessary and potentially dangerous procedures. We, therefore, set out to develop a simple, noninvasive, and quantitative assay to detect lung cancer. Methods: This proof-of-concept study evaluated the sensitivity/specificity of the CyPath Early Lung Cancer Detection Assay to correctly classify LDCT-confirmed cohorts of high-risk control (n = 102) and cancer (n = 26) subjects. Fluorescence intensity parameters of red fluorescent cells (RFCs) from tetra (4-carboxyphenyl) porphyrin (TCPP)-labeled lung sputum samples and subjects' baseline characteristics were assessed for their predictive power by multivariable logistic regression. A receiver operating characteristic curve was constructed to evaluate the sensitivity/specificity of the CyPath assay. Results: RFCs were detectable in cancer subjects more often than in high-risk ones (p = 0.015), and their characteristics differed between cohorts. Two independent predictors of cancer were the mean of RFC average fluorescence intensity/area per subject (p < 0.001) and years smoked (p = 0.003). The CyPath-based classifier had an overall accuracy of 81% in the test population; false-positive rate of 40% and negative predictive value of 83%. Conclusions: The tetra (4-carboxyphenyl) porphyrin -based CyPath assay correctly classified study participants into cancer or high-risk cohorts with considerable accuracy. Optimizing sputum collection, sample reading, and refining the classifier should improve sensitivity and specificity. The CyPath assay thus has the potential to complement LDCT screening or serve as a stand-alone approach for early lung cancer detection.
/EXPL THER/ Athymic nude mice with subcutaneously xenotransplanted urothelial carcinoma received intravenously injections of the synthetic sensitizer meso-tetra (4-carboxyphenyl) porphyrin. Fluorescence at the tumors and the skin was excited using a Kr+ laser (407 nm) and was detected at 652 nm using a fiberoptical sensor in combination with an optical multichannel analyzer at different times after application of the sensitizer. Photodynamic treatment was carried out using an Ar+ laser pumped dye laser (650 nm; 100 mW/sq cm; 100 J/sq cm). The delay of tumor growth was determined in a follow-up period of four weeks and was correlated to the in vivo fluorescence measurements.

C48H30N4O8

790.79

790.206

14609-54-2

86278368

Dark blue to purple to black powder

1.5±0.1 g/cm3

> 300 °C

1.734

11.46

6

10

8

60

1310

0

O([H])C(C1C([H])=C([H])C(=C([H])C=1[H])C1C2C([H])=C([H])C(=C(C3C([H])=C([H])C(C(=O)O[H])=C([H])C=3[H])C3=C([H])C([H])=C(C(C4C([H])=C([H])C(C(=O)O[H])=C([H])C=4[H])=C4C([H])=C([H])C(C(C5C([H])=C([H])C(C(=O)O[H])=C([H])C=5[H])=C5C([H])=C([H])C=1N5[H])=N4)N3[H])N=2)=O |c:20,86,t:58,80|

HHDUMDVQUCBCEY-UHFFFAOYSA-N

InChI=1S/C48H30N4O8/c53-45(54)29-9-1-25(2-10-29)41-33-17-19-35(49-33)42(26-3-11-30(12-4-26)46(55)56)37-21-23-39(51-37)44(28-7-15-32(16-8-28)48(59)60)40-24-22-38(52-40)43(36-20-18-34(41)50-36)27-5-13-31(14-6-27)47(57)58/h1-24,49,52H,(H,53,54)(H,55,56)(H,57,58)(H,59,60)

4-[10,15,20-tris(4-carboxyphenyl)-21,23-dihydroporphyrin-5-yl]benzoic acid

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Note: (1). This product requires protection from light (avoid light exposure) during transportation and storage.  (2). Please store this product in a sealed and protected environment (e.g. under nitrogen), avoid exposure to moisture.

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

DMSO: 4 mg/mL (5.06 mM)

Solubility in Formulation 1: 2.5 mg/mL (3.16 mM) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), suspension solution; with sonication.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL.
Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution.

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Solubility in Formulation 2: 2.5 mg/mL (3.16 mM) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), suspension solution; with ultrasonication.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly.
Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution.

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 (Please use freshly prepared in vivo formulations for optimal results.)