Journal of Drug Design and Medicinal Chemistry

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The Study of Cyclic Analogues of Carnosine Peptide on Isolated Mitochondria of A549 Cells

Received: Oct. 08, 2022    Accepted: Nov. 09, 2022    Published: Jan. 31, 2023
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Abstract

Bachground & Aims: In this work a various range of toxicity on A549 cells was resulted by the cyclic Carnosine analogues using MTT assay. The purpose of this research was to study the effects of analogues Carnosine peptide on A549 cells using several experiments. Also, applying peptides on the mitochondria of A549 cells, a raise of mitochondrial reactive oxygen species (ROS) level, mitochondrial swelling, mitochondrial membrane potential (ψm) collapse, release of cytochrome c of the affected mitochondria were detected. Methods: For determination of Cytotoxicity, Normal calls and A549 cancerous cells (1×107/well) were transferred into 96-well plates and treated with 10 μg/mL concentration of Carnosine analogues for 12h. The effect of peptides on the activity of SDH was assayed by MTT test. 100 μL mitochondrial suspensions from A549 and normal groups were incubated with applied concentration of peptides (10μg/mL) at 37°C for 30 min. The fluorescence intensity of DCFH which is an indicator of ROS concentration was then assayed by a Shimadzu RF-5000U fluorescence spectrophotometer. Mitochondrial accumulation and also redistribution of the cationic fluorescent dye, Rhoda mine 123 (Rh 123, concentration, 10 µM), from mitochondria into the cytosol have been used for the determination of MMP collapse. Mitochondria from A549 and normal groups were suspended in corresponding assay buffer and incubated at 37°C with 10µg/mL peptides. The release of Cytochrome c by peptides was assayed by the Quantikine Cytochrome c. Results: The results showed that all the synthesized cyclic peptides increased ROS in various levels in comparison with unaffected mitochondria isolated from A549 group. A significant increase of ROS was resulted by 2c, 4c and 6c analogues at 30 min. Rh 123 fluorescence staining indicated that the integrity of the mitochondria was damaged by the cyclic peptides. Compounds 2c, 4c and 6c significantly increased the collapse of MMP among the Carnosine analogues in comparison with mitochondria isolated from A549 group. Peptides 2c, 4c and 6c significantly increased mitochondrial swelling in comparison with untreated mitochondria isolated from the A549 group. The result was that cyclic peptides 2c, 4c and 6c significantly increased the release of cytochrome c in comparison with unaffected mitochondria isolated from the A549 group. Conclusion: Based on the overall results, cyclic analogues of Carnosine peptide, especially compounds 2c [Cyclo-(Pro-β-alanine-His-β-alanine-His)], 4c [Cyclo-(Pro-His-β-alanine-β-alanine-His)] and 6c [Cyclo-(Pro-β-alanine-His-His-β-alanine)], showed more toxic activity than other Carnosine analogues, which would be supporting to develop these peptide analogues as new anticancer and complementary therapeutic agents for the treatment of lung cancer.

DOI 10.11648/j.jddmc.20220804.12
Published in Journal of Drug Design and Medicinal Chemistry ( Volume 8, Issue 4, December 2022 )
Page(s) 50-56
Creative Commons

This is an Open Access article, distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution and reproduction in any medium or format, provided the original work is properly cited.

Copyright

Copyright © The Author(s), 2024. Published by Science Publishing Group

Keywords

Cyclic Analogues, Linear Analogues, Cytotoxicity, Mitochondria

References
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    Mohammadreza Gholibeikian, Nastaran Gholami Samali, Amirreza Arvaneh. (2023). The Study of Cyclic Analogues of Carnosine Peptide on Isolated Mitochondria of A549 Cells. Journal of Drug Design and Medicinal Chemistry, 8(4), 50-56. https://doi.org/10.11648/j.jddmc.20220804.12

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    Mohammadreza Gholibeikian; Nastaran Gholami Samali; Amirreza Arvaneh. The Study of Cyclic Analogues of Carnosine Peptide on Isolated Mitochondria of A549 Cells. J. Drug Des. Med. Chem. 2023, 8(4), 50-56. doi: 10.11648/j.jddmc.20220804.12

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    AMA Style

    Mohammadreza Gholibeikian, Nastaran Gholami Samali, Amirreza Arvaneh. The Study of Cyclic Analogues of Carnosine Peptide on Isolated Mitochondria of A549 Cells. J Drug Des Med Chem. 2023;8(4):50-56. doi: 10.11648/j.jddmc.20220804.12

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  • @article{10.11648/j.jddmc.20220804.12,
      author = {Mohammadreza Gholibeikian and Nastaran Gholami Samali and Amirreza Arvaneh},
      title = {The Study of Cyclic Analogues of Carnosine Peptide on Isolated Mitochondria of A549 Cells},
      journal = {Journal of Drug Design and Medicinal Chemistry},
      volume = {8},
      number = {4},
      pages = {50-56},
      doi = {10.11648/j.jddmc.20220804.12},
      url = {https://doi.org/10.11648/j.jddmc.20220804.12},
      eprint = {https://download.sciencepg.com/pdf/10.11648.j.jddmc.20220804.12},
      abstract = {Bachground & Aims: In this work a various range of toxicity on A549 cells was resulted by the cyclic Carnosine analogues using MTT assay. The purpose of this research was to study the effects of analogues Carnosine peptide on A549 cells using several experiments. Also, applying peptides on the mitochondria of A549 cells, a raise of mitochondrial reactive oxygen species (ROS) level, mitochondrial swelling, mitochondrial membrane potential (ψm) collapse, release of cytochrome c of the affected mitochondria were detected. Methods: For determination of Cytotoxicity, Normal calls and A549 cancerous cells (1×107/well) were transferred into 96-well plates and treated with 10 μg/mL concentration of Carnosine analogues for 12h. The effect of peptides on the activity of SDH was assayed by MTT test. 100 μL mitochondrial suspensions from A549 and normal groups were incubated with applied concentration of peptides (10μg/mL) at 37°C for 30 min. The fluorescence intensity of DCFH which is an indicator of ROS concentration was then assayed by a Shimadzu RF-5000U fluorescence spectrophotometer. Mitochondrial accumulation and also redistribution of the cationic fluorescent dye, Rhoda mine 123 (Rh 123, concentration, 10 µM), from mitochondria into the cytosol have been used for the determination of MMP collapse. Mitochondria from A549 and normal groups were suspended in corresponding assay buffer and incubated at 37°C with 10µg/mL peptides. The release of Cytochrome c by peptides was assayed by the Quantikine Cytochrome c. Results: The results showed that all the synthesized cyclic peptides increased ROS in various levels in comparison with unaffected mitochondria isolated from A549 group. A significant increase of ROS was resulted by 2c, 4c and 6c analogues at 30 min. Rh 123 fluorescence staining indicated that the integrity of the mitochondria was damaged by the cyclic peptides. Compounds 2c, 4c and 6c significantly increased the collapse of MMP among the Carnosine analogues in comparison with mitochondria isolated from A549 group. Peptides 2c, 4c and 6c significantly increased mitochondrial swelling in comparison with untreated mitochondria isolated from the A549 group. The result was that cyclic peptides 2c, 4c and 6c significantly increased the release of cytochrome c in comparison with unaffected mitochondria isolated from the A549 group. Conclusion: Based on the overall results, cyclic analogues of Carnosine peptide, especially compounds 2c [Cyclo-(Pro-β-alanine-His-β-alanine-His)], 4c [Cyclo-(Pro-His-β-alanine-β-alanine-His)] and 6c [Cyclo-(Pro-β-alanine-His-His-β-alanine)], showed more toxic activity than other Carnosine analogues, which would be supporting to develop these peptide analogues as new anticancer and complementary therapeutic agents for the treatment of lung cancer.},
     year = {2023}
    }
    

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  • TY  - JOUR
    T1  - The Study of Cyclic Analogues of Carnosine Peptide on Isolated Mitochondria of A549 Cells
    AU  - Mohammadreza Gholibeikian
    AU  - Nastaran Gholami Samali
    AU  - Amirreza Arvaneh
    Y1  - 2023/01/31
    PY  - 2023
    N1  - https://doi.org/10.11648/j.jddmc.20220804.12
    DO  - 10.11648/j.jddmc.20220804.12
    T2  - Journal of Drug Design and Medicinal Chemistry
    JF  - Journal of Drug Design and Medicinal Chemistry
    JO  - Journal of Drug Design and Medicinal Chemistry
    SP  - 50
    EP  - 56
    PB  - Science Publishing Group
    SN  - 2472-3576
    UR  - https://doi.org/10.11648/j.jddmc.20220804.12
    AB  - Bachground & Aims: In this work a various range of toxicity on A549 cells was resulted by the cyclic Carnosine analogues using MTT assay. The purpose of this research was to study the effects of analogues Carnosine peptide on A549 cells using several experiments. Also, applying peptides on the mitochondria of A549 cells, a raise of mitochondrial reactive oxygen species (ROS) level, mitochondrial swelling, mitochondrial membrane potential (ψm) collapse, release of cytochrome c of the affected mitochondria were detected. Methods: For determination of Cytotoxicity, Normal calls and A549 cancerous cells (1×107/well) were transferred into 96-well plates and treated with 10 μg/mL concentration of Carnosine analogues for 12h. The effect of peptides on the activity of SDH was assayed by MTT test. 100 μL mitochondrial suspensions from A549 and normal groups were incubated with applied concentration of peptides (10μg/mL) at 37°C for 30 min. The fluorescence intensity of DCFH which is an indicator of ROS concentration was then assayed by a Shimadzu RF-5000U fluorescence spectrophotometer. Mitochondrial accumulation and also redistribution of the cationic fluorescent dye, Rhoda mine 123 (Rh 123, concentration, 10 µM), from mitochondria into the cytosol have been used for the determination of MMP collapse. Mitochondria from A549 and normal groups were suspended in corresponding assay buffer and incubated at 37°C with 10µg/mL peptides. The release of Cytochrome c by peptides was assayed by the Quantikine Cytochrome c. Results: The results showed that all the synthesized cyclic peptides increased ROS in various levels in comparison with unaffected mitochondria isolated from A549 group. A significant increase of ROS was resulted by 2c, 4c and 6c analogues at 30 min. Rh 123 fluorescence staining indicated that the integrity of the mitochondria was damaged by the cyclic peptides. Compounds 2c, 4c and 6c significantly increased the collapse of MMP among the Carnosine analogues in comparison with mitochondria isolated from A549 group. Peptides 2c, 4c and 6c significantly increased mitochondrial swelling in comparison with untreated mitochondria isolated from the A549 group. The result was that cyclic peptides 2c, 4c and 6c significantly increased the release of cytochrome c in comparison with unaffected mitochondria isolated from the A549 group. Conclusion: Based on the overall results, cyclic analogues of Carnosine peptide, especially compounds 2c [Cyclo-(Pro-β-alanine-His-β-alanine-His)], 4c [Cyclo-(Pro-His-β-alanine-β-alanine-His)] and 6c [Cyclo-(Pro-β-alanine-His-His-β-alanine)], showed more toxic activity than other Carnosine analogues, which would be supporting to develop these peptide analogues as new anticancer and complementary therapeutic agents for the treatment of lung cancer.
    VL  - 8
    IS  - 4
    ER  - 

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Author Information
  • Department of Organic Chemistry, Faculty of Chemistry, University of Kashan, Kashan, I.R. Iran

  • Department of Analytical Chemistry, Faculty of Chemistry, Shahid Chamran University of Ahvaz, Ahvaz, Iran

  • Department of Organic Chemistry, Faculty of Chemistry, University of Science and Technology, Tehran, I.R. Iran

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