Chemoselective Synthesis of Mannich Adducts from 1,4-Naphthoquinones and Profile as Autophagic Inducers in Oral Squamous Cell Carcinoma
Oral squamous cell carcinoma (OSCC) is a worldwide public health problem, accounting for approximately 90% of all oral cancers, and is the eighth most common cancer in men. Cisplatin and carboplatin are the main chemotherapy drugs used in the clinic. However, in addition to their serious side effect...
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MDPI AG
2022-12-01
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| Series: | Molecules |
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| Online Access: | https://www.mdpi.com/1420-3049/28/1/309 |
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| author | Amanda A. Borges Michele P. de Souza Anna Carolina C. da Fonseca Guilherme F. Wermelinger Ruan C. B. Ribeiro Adriane A. P. Amaral Cláudio José C. de Carvalho Lucas S. Abreu Lucas Nicolau de Queiroz Elan C. P. de Almeida Vitor W. Rabelo Paula A. Abreu Bruno Pontes Vitor F. Ferreira Fernando de C. da Silva Luana da S. M. Forezi Bruno K. Robbs |
| author_facet | Amanda A. Borges Michele P. de Souza Anna Carolina C. da Fonseca Guilherme F. Wermelinger Ruan C. B. Ribeiro Adriane A. P. Amaral Cláudio José C. de Carvalho Lucas S. Abreu Lucas Nicolau de Queiroz Elan C. P. de Almeida Vitor W. Rabelo Paula A. Abreu Bruno Pontes Vitor F. Ferreira Fernando de C. da Silva Luana da S. M. Forezi Bruno K. Robbs |
| author_sort | Amanda A. Borges |
| collection | DOAJ |
| description | Oral squamous cell carcinoma (OSCC) is a worldwide public health problem, accounting for approximately 90% of all oral cancers, and is the eighth most common cancer in men. Cisplatin and carboplatin are the main chemotherapy drugs used in the clinic. However, in addition to their serious side effects, such as damage to the nervous system and kidneys, there is also drug resistance. Thus, the development of new drugs becomes of great importance. Naphthoquinones have been described with antitumor activity. Some of them are found in nature, but semi synthesis has been used as strategy to find new chemical entities for the treatment of cancer. In the present study, we promote a multiple component reaction (MCR) among lawsone, arylaldehydes, and benzylamine to produce sixteen chemoselectively derivated Mannich adducts of 1,4-naphthoquinones in good yield (up to 97%). The antitumor activities and molecular mechanisms of action of these compounds were investigated in OSCC models and the compound <b>6a</b> induced cytotoxicity in three different tumor cell lines (OSCC4, OSCC9, and OSCC25) and was more selective (IS > 2) for tumor cells than the chemotropic drug carboplatin and the controls lapachol and shikonin, which are chemically similar compounds with cytotoxic effects. The <b>6a</b> selectively and significantly reduced the amount of cell colony growth, was not hemolytic, and tolerable in mice with no serious side effects at a concentration of 100 mg/kg with a LD<sub>50</sub> of 150 mg/kg. The new compound is biologically stable with a profile similar to carboplatin. Morphologically, <b>6a</b> does not induce cell retraction or membrane blebs, but it does induce intense vesicle formation and late emergence of membrane bubbles. Exploring the mechanism of cell death induction, compound <b>6a</b> does not induce ROS formation, and cell viability was not affected by inhibitors of apoptosis (ZVAD) and necroptosis (necrostatin 1). Autophagy followed by a late apoptosis process appears to be the death-inducing pathway of <b>6a</b>, as observed by increased viability by the autophagy inhibitor (3-MA) and by the appearance of autophagosomes, later triggering a process of late apoptosis with the presence of caspase 3/7 and DNA fragmentation. Molecular modeling suggests the ability of the compound to bind to topoisomerase I and II and with greater affinity to hPKM2 enzyme than controls, which could explain the mechanism of cell death by autophagy. Finally, the in-silico prediction of drug-relevant properties showed that compound <b>6a</b> has a good pharmacokinetic profile when compared to carboplatin and doxorubicin. Among the sixteen naphthoquinones tested, compound <b>6a</b> was the most effective and is highly selective and well tolerated in animals. The induction of cell death in OSCC through autophagy followed by late apoptosis possibly via inhibition of the PKM2 enzyme points to a promising potential of <b>6a</b> as a new preclinical anticancer candidate. |
| first_indexed | 2024-03-09T12:05:52Z |
| format | Article |
| id | doaj.art-91cc9dcabd21467cb4e362422dc6a0af |
| institution | Directory Open Access Journal |
| issn | 1420-3049 |
| language | English |
| last_indexed | 2024-03-09T12:05:52Z |
| publishDate | 2022-12-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Molecules |
| spelling | doaj.art-91cc9dcabd21467cb4e362422dc6a0af2023-11-30T22:57:51ZengMDPI AGMolecules1420-30492022-12-0128130910.3390/molecules28010309Chemoselective Synthesis of Mannich Adducts from 1,4-Naphthoquinones and Profile as Autophagic Inducers in Oral Squamous Cell CarcinomaAmanda A. Borges0Michele P. de Souza1Anna Carolina C. da Fonseca2Guilherme F. Wermelinger3Ruan C. B. Ribeiro4Adriane A. P. Amaral5Cláudio José C. de Carvalho6Lucas S. Abreu7Lucas Nicolau de Queiroz8Elan C. P. de Almeida9Vitor W. Rabelo10Paula A. Abreu11Bruno Pontes12Vitor F. Ferreira13Fernando de C. da Silva14Luana da S. M. Forezi15Bruno K. Robbs16Departamento de Química Orgânica, Instituto de Química, Campus do Valonguinho, Universidade Federal Fluminense, Niterói CEP 24020-150, BrazilPrograma de Pós-Graduação em Ciências Aplicadas a Produtos para Saúde, Faculdade de Farmácia, Universidade Federal Fluminense, Niterói CEP 24241-000, BrazilPrograma de Pós-Graduação em Odontologia, Instituto de Saúde de Nova Friburgo, Universidade Federal Fluminense, Nova Friburgo CEP 28625-650, BrazilDepartamento de Ciência Básica, Campus Universitário de Nova Friburgo, Universidade Federal Fluminense, Nova Friburgo CEP 28625-650, BrazilDepartamento de Química Orgânica, Instituto de Química, Campus do Valonguinho, Universidade Federal Fluminense, Niterói CEP 24020-150, BrazilDepartamento de Química Orgânica, Instituto de Química, Campus do Valonguinho, Universidade Federal Fluminense, Niterói CEP 24020-150, BrazilDepartamento de Química Orgânica, Instituto de Química, Campus do Valonguinho, Universidade Federal Fluminense, Niterói CEP 24020-150, BrazilDepartamento de Química Orgânica, Instituto de Química, Campus do Valonguinho, Universidade Federal Fluminense, Niterói CEP 24020-150, BrazilPrograma de Pós-Graduação em Ciências Aplicadas a Produtos para Saúde, Faculdade de Farmácia, Universidade Federal Fluminense, Niterói CEP 24241-000, BrazilDepartamento de Ciência Básica, Campus Universitário de Nova Friburgo, Universidade Federal Fluminense, Nova Friburgo CEP 28625-650, BrazilInstituto de Biodiversidade e Sustentabilidade, Campus Macaé, Universidade Federal do Rio de Janeiro, Macaé CEP 27965-045, BrazilInstituto de Biodiversidade e Sustentabilidade, Campus Macaé, Universidade Federal do Rio de Janeiro, Macaé CEP 27965-045, BrazilInstituto de Ciências Biomédicas, Universidade Federal do Rio de Janeiro, Rio de Janeiro CEP 21941-902, BrazilDepartamento de Tecnologia Farmacêutica, Faculdade de Farmácia, Universidade Federal Fluminense, Niterói CEP 24241-000, BrazilDepartamento de Química Orgânica, Instituto de Química, Campus do Valonguinho, Universidade Federal Fluminense, Niterói CEP 24020-150, BrazilDepartamento de Química Orgânica, Instituto de Química, Campus do Valonguinho, Universidade Federal Fluminense, Niterói CEP 24020-150, BrazilDepartamento de Química Orgânica, Instituto de Química, Campus do Valonguinho, Universidade Federal Fluminense, Niterói CEP 24020-150, BrazilOral squamous cell carcinoma (OSCC) is a worldwide public health problem, accounting for approximately 90% of all oral cancers, and is the eighth most common cancer in men. Cisplatin and carboplatin are the main chemotherapy drugs used in the clinic. However, in addition to their serious side effects, such as damage to the nervous system and kidneys, there is also drug resistance. Thus, the development of new drugs becomes of great importance. Naphthoquinones have been described with antitumor activity. Some of them are found in nature, but semi synthesis has been used as strategy to find new chemical entities for the treatment of cancer. In the present study, we promote a multiple component reaction (MCR) among lawsone, arylaldehydes, and benzylamine to produce sixteen chemoselectively derivated Mannich adducts of 1,4-naphthoquinones in good yield (up to 97%). The antitumor activities and molecular mechanisms of action of these compounds were investigated in OSCC models and the compound <b>6a</b> induced cytotoxicity in three different tumor cell lines (OSCC4, OSCC9, and OSCC25) and was more selective (IS > 2) for tumor cells than the chemotropic drug carboplatin and the controls lapachol and shikonin, which are chemically similar compounds with cytotoxic effects. The <b>6a</b> selectively and significantly reduced the amount of cell colony growth, was not hemolytic, and tolerable in mice with no serious side effects at a concentration of 100 mg/kg with a LD<sub>50</sub> of 150 mg/kg. The new compound is biologically stable with a profile similar to carboplatin. Morphologically, <b>6a</b> does not induce cell retraction or membrane blebs, but it does induce intense vesicle formation and late emergence of membrane bubbles. Exploring the mechanism of cell death induction, compound <b>6a</b> does not induce ROS formation, and cell viability was not affected by inhibitors of apoptosis (ZVAD) and necroptosis (necrostatin 1). Autophagy followed by a late apoptosis process appears to be the death-inducing pathway of <b>6a</b>, as observed by increased viability by the autophagy inhibitor (3-MA) and by the appearance of autophagosomes, later triggering a process of late apoptosis with the presence of caspase 3/7 and DNA fragmentation. Molecular modeling suggests the ability of the compound to bind to topoisomerase I and II and with greater affinity to hPKM2 enzyme than controls, which could explain the mechanism of cell death by autophagy. Finally, the in-silico prediction of drug-relevant properties showed that compound <b>6a</b> has a good pharmacokinetic profile when compared to carboplatin and doxorubicin. Among the sixteen naphthoquinones tested, compound <b>6a</b> was the most effective and is highly selective and well tolerated in animals. The induction of cell death in OSCC through autophagy followed by late apoptosis possibly via inhibition of the PKM2 enzyme points to a promising potential of <b>6a</b> as a new preclinical anticancer candidate.https://www.mdpi.com/1420-3049/28/1/309oral squamous cell carcinomalawsonelapacholMannich adductspyruvate kinase M2autophagy |
| spellingShingle | Amanda A. Borges Michele P. de Souza Anna Carolina C. da Fonseca Guilherme F. Wermelinger Ruan C. B. Ribeiro Adriane A. P. Amaral Cláudio José C. de Carvalho Lucas S. Abreu Lucas Nicolau de Queiroz Elan C. P. de Almeida Vitor W. Rabelo Paula A. Abreu Bruno Pontes Vitor F. Ferreira Fernando de C. da Silva Luana da S. M. Forezi Bruno K. Robbs Chemoselective Synthesis of Mannich Adducts from 1,4-Naphthoquinones and Profile as Autophagic Inducers in Oral Squamous Cell Carcinoma Molecules oral squamous cell carcinoma lawsone lapachol Mannich adducts pyruvate kinase M2 autophagy |
| title | Chemoselective Synthesis of Mannich Adducts from 1,4-Naphthoquinones and Profile as Autophagic Inducers in Oral Squamous Cell Carcinoma |
| title_full | Chemoselective Synthesis of Mannich Adducts from 1,4-Naphthoquinones and Profile as Autophagic Inducers in Oral Squamous Cell Carcinoma |
| title_fullStr | Chemoselective Synthesis of Mannich Adducts from 1,4-Naphthoquinones and Profile as Autophagic Inducers in Oral Squamous Cell Carcinoma |
| title_full_unstemmed | Chemoselective Synthesis of Mannich Adducts from 1,4-Naphthoquinones and Profile as Autophagic Inducers in Oral Squamous Cell Carcinoma |
| title_short | Chemoselective Synthesis of Mannich Adducts from 1,4-Naphthoquinones and Profile as Autophagic Inducers in Oral Squamous Cell Carcinoma |
| title_sort | chemoselective synthesis of mannich adducts from 1 4 naphthoquinones and profile as autophagic inducers in oral squamous cell carcinoma |
| topic | oral squamous cell carcinoma lawsone lapachol Mannich adducts pyruvate kinase M2 autophagy |
| url | https://www.mdpi.com/1420-3049/28/1/309 |
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