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Simultaneous Dual Selective Targeted Delivery of Two Covalent Gemcitabine Immunochemotherapeutics and Complementary Anti-Neoplastic Potency of [Se]-Methylselenocysteine.

Coyne CP, Jones T, Bear R - J Cancer Ther (2015)

Bottom Line: Gemcitabine-(C4-amide)-[anti-EGFR] and gemcitabine-(C4-amide)-[anti-HER2/neu] produced progressive increases in anti-neoplastic cytotoxicity that were greatest between gemcitabine-equivalent concentrations of 10(-9) M and 10(-6) M.Dual simultaneous combinations of gemcitabine-(C4-amide)-[anti-EGFR] with gemcitabine-(C4-amide)-[anti-HER2/neu] produced levels of anti-neoplastic cytotoxicity intermediate between each of the individual covalent gemcitabine immunochemotherapeutics.Total anti-neoplastic cytotoxicity of the dual simultaneous combination of gemcitabine-(C4-amide)-[anti-EGFR] and gemcitabine-(C4-amide)-[anti-HER2/neu] against chemotherapeutic-resistant mammary adenocarcinoma (SKBr-3) was substantially higher when formulated with [Se]-methylsele-nocysteine.

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Affiliation: Department of Basic Sciences, College of Veterinary Medicine, Mississippi State University, Mississippi State, USA.

ABSTRACT

The anti-metabolite chemotherapeutic, gemcitabine is relatively effective for a spectrum of neoplastic conditions that include various forms of leukemia and adenocarcinoma/carcinoma. Rapid systemic deamination of gemcitabine accounts for a brief plasma half-life but its sustained administration is often curtailed by sequelae and chemotherapeutic-resistance. A molecular strategy that diminishes these limitations is the molecular design and synthetic production of covalent gemcitabine immunochemotherapeutics that possess properties of selective "targeted" delivery. The simultaneous dual selective "targeted" delivery of gemcitabine at two separate sites on the external surface membrane of a single cancer cell types represents a therapeutic approach that can increase cytosol chemotherapeutic deposition; prolong chemotherapeutic plasma half-life (reduces administration frequency); minimize innocent exposure of normal tissues and healthy organ systems; and ultimately enhance more rapid and thorough resolution of neoplastic cell populations.

Materials and methods: A light-reactive gemcitabine intermediate synthesized utilizing succinimidyl 4,4-azipentanoate was covalently bound to anti-EGFR or anti-HER2/neu IgG by exposure to UV light (354-nm) resulting in the synthesis of covalent immunochemotherapeutics, gemcitabine-(C4-amide)-[anti-EGFR] and gemcitabine-(C4-amide)-[anti-HER2/neu]. Cytotoxic anti-neoplastic potency of gemcitabine-(C4-amide)-[anti-EGFR] and gemcitabine-(C4-amide)-[anti-HER2/neu] between gemcitabine-equivalent concentrations of 10(-12) M and 10(-6) M was determined utilizing chemotherapeutic-resistant mammary adenocarcinoma (SKRr-3). The organoselenium compound, [Se]-methylselenocysteine was evaluated to determine if it complemented the anti-neoplastic potency of the covalent gemcitabine immunochemotherapeutics.

Results: Gemcitabine-(C4-amide)-[anti-EGFR], gemcitabine-(C4-amide)-[anti-HER2/neu] and the dual simultaneous combination of gemcitabine-(C4-amide)-[anti-EGFR] with gemcitabine-(C4-amide)-[anti-HER2/neu] all had anti-neoplastic cytotoxic potency against mammary adenocarcinoma. Gemcitabine-(C4-amide)-[anti-EGFR] and gemcitabine-(C4-amide)-[anti-HER2/neu] produced progressive increases in anti-neoplastic cytotoxicity that were greatest between gemcitabine-equivalent concentrations of 10(-9) M and 10(-6) M. Dual simultaneous combinations of gemcitabine-(C4-amide)-[anti-EGFR] with gemcitabine-(C4-amide)-[anti-HER2/neu] produced levels of anti-neoplastic cytotoxicity intermediate between each of the individual covalent gemcitabine immunochemotherapeutics. Total anti-neoplastic cytotoxicity of the dual simultaneous combination of gemcitabine-(C4-amide)-[anti-EGFR] and gemcitabine-(C4-amide)-[anti-HER2/neu] against chemotherapeutic-resistant mammary adenocarcinoma (SKBr-3) was substantially higher when formulated with [Se]-methylsele-nocysteine.

No MeSH data available.


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Relative anti-neoplastic cytotoxicity for dual simultaneous combinations of two different covalent gemcitabine-immunochemotherapeutics enhanced by [Se]-methylselenocysteine against chemotherapeutic-resistant human mammary adenocarcinoma. Legends: (◆) gemcitabine-(C4-amide)-[anti-EGFR] with gemcitabine-(C4-amide)-[anti-HER2/neu]; and (■) gemcitabine-(C4-amide)-[anti-EGFR] with gemcitabine-(C4-amide)-[anti-HER2/neu] in the presence of a fixed concentration of [Se]-methyl- cysteine (15 μM). The dual simultaneous combination of covalent gemcitabine-immunochemotherapeutics (+/− [Se]-methylcysteine) was formulated at gradient 50/50 gemcitabine-equivalent concentrations and incubated in direct contact for 96-hours with triplicate monolayer populations of chemotherapeutic-resistant human mammary adenocarcinoma (SKBr-3). Anti-neoplastic cytotoxicity was measured using a MTT cell vitality assay relative to matched negative reference controls.
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Figure 10: Relative anti-neoplastic cytotoxicity for dual simultaneous combinations of two different covalent gemcitabine-immunochemotherapeutics enhanced by [Se]-methylselenocysteine against chemotherapeutic-resistant human mammary adenocarcinoma. Legends: (◆) gemcitabine-(C4-amide)-[anti-EGFR] with gemcitabine-(C4-amide)-[anti-HER2/neu]; and (■) gemcitabine-(C4-amide)-[anti-EGFR] with gemcitabine-(C4-amide)-[anti-HER2/neu] in the presence of a fixed concentration of [Se]-methyl- cysteine (15 μM). The dual simultaneous combination of covalent gemcitabine-immunochemotherapeutics (+/− [Se]-methylcysteine) was formulated at gradient 50/50 gemcitabine-equivalent concentrations and incubated in direct contact for 96-hours with triplicate monolayer populations of chemotherapeutic-resistant human mammary adenocarcinoma (SKBr-3). Anti-neoplastic cytotoxicity was measured using a MTT cell vitality assay relative to matched negative reference controls.

Mentions: Methylseleninate produced levels of anti-neoplastic cytotoxicity that were substantially greater than those detected for [Se]-methylselenocysteine at the selenium-equivalent concentrations of 10 μM, and 20 μM but approached similar levels at 30 μM and 40 μM with essentially equivalent potency observed at 50 mM respectively (Figure 9). Methylseninate had almost equivalent maximal levels of anti-neoplastic cytotoxicity at and between the selenium-equivalent concentrations of 10 μM and 50 μM while [Se]-methylselenocysteine produced rapid progressive increases in anti-neoplastic cytotoxicity at and between 10 μM and 30 μM while levels were near maximum at 30 μM, 20 μM and 10 μM (Figure 9). [Se]-methylselenocysteine substantially contributed to the anti-neoplastic cytotoxicity of gemcitabine-standardized 50/50 formulations of gemcitabine-(C4-amide)-[anti-EGFR] with gemcitabine-(C4-amide)-[anti-HER2/neu] compared to only the dual simultaneous combination of the two covalent gemcitabine immunochemotherapeutics (Figure 8 and Figure 10). Gemcitabine-(C4-amide)-[anti-EGFR] and gemcitabine-(C4-amide)-[anti-HER2/neu] produced progressive increases in anti-neoplastic cytotoxicity that were most rapid at and between the gemcitabine-equivalent concentrations of 10−9 M and 10−6 M (Figure 8 and Figure 10). [Se]-methylselenocysteine (15 μM final concentration) in combination with the two covalent gemcitabine immunochemotherapeutics resulted in anti-neoplastic cytotoxicity levels of 93.5% at 10−10 M (6.5% residual survival), 93.9% at 10−9 M (6.1% residual survival), 94.7% at 10−8 M (5.3% residual survival), 94.2% at 10−7 M (5.8% residual survival), and 94.2% at 10−6 M (5.8% residual survival) following a direct-contact incubation period (Figure 10).


Simultaneous Dual Selective Targeted Delivery of Two Covalent Gemcitabine Immunochemotherapeutics and Complementary Anti-Neoplastic Potency of [Se]-Methylselenocysteine.

Coyne CP, Jones T, Bear R - J Cancer Ther (2015)

Relative anti-neoplastic cytotoxicity for dual simultaneous combinations of two different covalent gemcitabine-immunochemotherapeutics enhanced by [Se]-methylselenocysteine against chemotherapeutic-resistant human mammary adenocarcinoma. Legends: (◆) gemcitabine-(C4-amide)-[anti-EGFR] with gemcitabine-(C4-amide)-[anti-HER2/neu]; and (■) gemcitabine-(C4-amide)-[anti-EGFR] with gemcitabine-(C4-amide)-[anti-HER2/neu] in the presence of a fixed concentration of [Se]-methyl- cysteine (15 μM). The dual simultaneous combination of covalent gemcitabine-immunochemotherapeutics (+/− [Se]-methylcysteine) was formulated at gradient 50/50 gemcitabine-equivalent concentrations and incubated in direct contact for 96-hours with triplicate monolayer populations of chemotherapeutic-resistant human mammary adenocarcinoma (SKBr-3). Anti-neoplastic cytotoxicity was measured using a MTT cell vitality assay relative to matched negative reference controls.
© Copyright Policy - open-access
Related In: Results  -  Collection

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Show All Figures
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Figure 10: Relative anti-neoplastic cytotoxicity for dual simultaneous combinations of two different covalent gemcitabine-immunochemotherapeutics enhanced by [Se]-methylselenocysteine against chemotherapeutic-resistant human mammary adenocarcinoma. Legends: (◆) gemcitabine-(C4-amide)-[anti-EGFR] with gemcitabine-(C4-amide)-[anti-HER2/neu]; and (■) gemcitabine-(C4-amide)-[anti-EGFR] with gemcitabine-(C4-amide)-[anti-HER2/neu] in the presence of a fixed concentration of [Se]-methyl- cysteine (15 μM). The dual simultaneous combination of covalent gemcitabine-immunochemotherapeutics (+/− [Se]-methylcysteine) was formulated at gradient 50/50 gemcitabine-equivalent concentrations and incubated in direct contact for 96-hours with triplicate monolayer populations of chemotherapeutic-resistant human mammary adenocarcinoma (SKBr-3). Anti-neoplastic cytotoxicity was measured using a MTT cell vitality assay relative to matched negative reference controls.
Mentions: Methylseleninate produced levels of anti-neoplastic cytotoxicity that were substantially greater than those detected for [Se]-methylselenocysteine at the selenium-equivalent concentrations of 10 μM, and 20 μM but approached similar levels at 30 μM and 40 μM with essentially equivalent potency observed at 50 mM respectively (Figure 9). Methylseninate had almost equivalent maximal levels of anti-neoplastic cytotoxicity at and between the selenium-equivalent concentrations of 10 μM and 50 μM while [Se]-methylselenocysteine produced rapid progressive increases in anti-neoplastic cytotoxicity at and between 10 μM and 30 μM while levels were near maximum at 30 μM, 20 μM and 10 μM (Figure 9). [Se]-methylselenocysteine substantially contributed to the anti-neoplastic cytotoxicity of gemcitabine-standardized 50/50 formulations of gemcitabine-(C4-amide)-[anti-EGFR] with gemcitabine-(C4-amide)-[anti-HER2/neu] compared to only the dual simultaneous combination of the two covalent gemcitabine immunochemotherapeutics (Figure 8 and Figure 10). Gemcitabine-(C4-amide)-[anti-EGFR] and gemcitabine-(C4-amide)-[anti-HER2/neu] produced progressive increases in anti-neoplastic cytotoxicity that were most rapid at and between the gemcitabine-equivalent concentrations of 10−9 M and 10−6 M (Figure 8 and Figure 10). [Se]-methylselenocysteine (15 μM final concentration) in combination with the two covalent gemcitabine immunochemotherapeutics resulted in anti-neoplastic cytotoxicity levels of 93.5% at 10−10 M (6.5% residual survival), 93.9% at 10−9 M (6.1% residual survival), 94.7% at 10−8 M (5.3% residual survival), 94.2% at 10−7 M (5.8% residual survival), and 94.2% at 10−6 M (5.8% residual survival) following a direct-contact incubation period (Figure 10).

Bottom Line: Gemcitabine-(C4-amide)-[anti-EGFR] and gemcitabine-(C4-amide)-[anti-HER2/neu] produced progressive increases in anti-neoplastic cytotoxicity that were greatest between gemcitabine-equivalent concentrations of 10(-9) M and 10(-6) M.Dual simultaneous combinations of gemcitabine-(C4-amide)-[anti-EGFR] with gemcitabine-(C4-amide)-[anti-HER2/neu] produced levels of anti-neoplastic cytotoxicity intermediate between each of the individual covalent gemcitabine immunochemotherapeutics.Total anti-neoplastic cytotoxicity of the dual simultaneous combination of gemcitabine-(C4-amide)-[anti-EGFR] and gemcitabine-(C4-amide)-[anti-HER2/neu] against chemotherapeutic-resistant mammary adenocarcinoma (SKBr-3) was substantially higher when formulated with [Se]-methylsele-nocysteine.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Basic Sciences, College of Veterinary Medicine, Mississippi State University, Mississippi State, USA.

ABSTRACT

The anti-metabolite chemotherapeutic, gemcitabine is relatively effective for a spectrum of neoplastic conditions that include various forms of leukemia and adenocarcinoma/carcinoma. Rapid systemic deamination of gemcitabine accounts for a brief plasma half-life but its sustained administration is often curtailed by sequelae and chemotherapeutic-resistance. A molecular strategy that diminishes these limitations is the molecular design and synthetic production of covalent gemcitabine immunochemotherapeutics that possess properties of selective "targeted" delivery. The simultaneous dual selective "targeted" delivery of gemcitabine at two separate sites on the external surface membrane of a single cancer cell types represents a therapeutic approach that can increase cytosol chemotherapeutic deposition; prolong chemotherapeutic plasma half-life (reduces administration frequency); minimize innocent exposure of normal tissues and healthy organ systems; and ultimately enhance more rapid and thorough resolution of neoplastic cell populations.

Materials and methods: A light-reactive gemcitabine intermediate synthesized utilizing succinimidyl 4,4-azipentanoate was covalently bound to anti-EGFR or anti-HER2/neu IgG by exposure to UV light (354-nm) resulting in the synthesis of covalent immunochemotherapeutics, gemcitabine-(C4-amide)-[anti-EGFR] and gemcitabine-(C4-amide)-[anti-HER2/neu]. Cytotoxic anti-neoplastic potency of gemcitabine-(C4-amide)-[anti-EGFR] and gemcitabine-(C4-amide)-[anti-HER2/neu] between gemcitabine-equivalent concentrations of 10(-12) M and 10(-6) M was determined utilizing chemotherapeutic-resistant mammary adenocarcinoma (SKRr-3). The organoselenium compound, [Se]-methylselenocysteine was evaluated to determine if it complemented the anti-neoplastic potency of the covalent gemcitabine immunochemotherapeutics.

Results: Gemcitabine-(C4-amide)-[anti-EGFR], gemcitabine-(C4-amide)-[anti-HER2/neu] and the dual simultaneous combination of gemcitabine-(C4-amide)-[anti-EGFR] with gemcitabine-(C4-amide)-[anti-HER2/neu] all had anti-neoplastic cytotoxic potency against mammary adenocarcinoma. Gemcitabine-(C4-amide)-[anti-EGFR] and gemcitabine-(C4-amide)-[anti-HER2/neu] produced progressive increases in anti-neoplastic cytotoxicity that were greatest between gemcitabine-equivalent concentrations of 10(-9) M and 10(-6) M. Dual simultaneous combinations of gemcitabine-(C4-amide)-[anti-EGFR] with gemcitabine-(C4-amide)-[anti-HER2/neu] produced levels of anti-neoplastic cytotoxicity intermediate between each of the individual covalent gemcitabine immunochemotherapeutics. Total anti-neoplastic cytotoxicity of the dual simultaneous combination of gemcitabine-(C4-amide)-[anti-EGFR] and gemcitabine-(C4-amide)-[anti-HER2/neu] against chemotherapeutic-resistant mammary adenocarcinoma (SKBr-3) was substantially higher when formulated with [Se]-methylsele-nocysteine.

No MeSH data available.


Related in: MedlinePlus