Combination of Conventional Drugs with Biocompounds Derived from Cinnamic Acid: A Promising Option for Breast Cancer Therapy.

Despite the options available for breast cancer (BC) therapy, several adverse effects and resistance limit the success of the treatment. Furthermore, the use of a single drug is associated with a high failure rate. We investigated through a systematic review the in vitro effects of the combination between conventional drugs and bioactive compounds derived from cinnamic acid in BC treatment. The information was acquired from the following databases: PubMed, Web of Science, Embase, Scopus, Lilacs and Cochrane library. We focused on "Cinnamates", "Drug Combinations" and "Breast neoplasms" for publications dating between January 2012 and December 2022, based on the PRISMA statement. The references of the articles were carefully reviewed. Finally, nine eligible studies were included. The majority of these studies were performed using MCF-7, MDA-MB-231, MDA-MB-468 and BT-20 cell lines and the combination between cisplatin, paclitaxel, doxorubicin, tamoxifen, dactolisib and veliparib, with caffeic acid phenethyl ester, eugenol, 3-caffeoylquinic acid, salvianolic acid A, ferulic acid, caffeic acid, rosmarinic acid and ursolic acid. The combination improved overall conventional drug effects, with increased cytotoxicity, antimigratory effect and reversing resistance. Combining conventional drugs with bioactive compounds derived from cinnamic acid could emerge as a privileged scaffold for establishing new treatment options for different BC types.

In vitro evidence that the anti-inflammatory effect of synthetic cinnamate-derived dienes is directly linked to a macrophage repolarization.

The inflammatory process is a mammalian physiological reaction against infectious agents or injuries. Among the cells involved, the macrophages have a highlighted role during this process. Depending on the inflammatory context, they can polarize into pro- or anti-inflammatory profiles (M1 and M2). In this context, compounds derived from cinnamic acid have demonstrated strong evidence of anti-inflammatory activity; however, the mechanism responsible for this effect remains unclear. In this study, we investigated the anti-inflammatory activity of five cinnamate-derived dienes of synthetic origin. The compounds that did not demonstrate significant cytotoxicity were tested to assess anti-inflammatory activity (NOx ) in RAW 264.7 cells stimulated with LPS. Then, the selected compound (diene 1) was evaluated as to its ability to inhibit the secretion of pro-inflammatory cytokines (IL-1ß, TNF-α, INF-γ, MCP-1, and IL-6) and increase the production of anti-inflammatory cytokines (IL-13, IL-4, and IL-10). Finally, diene 1 was able to reduce the expression of TLR4 and increase the phagocytic activity of the macrophages. Gathering these results together, we conclude that diene 1 showed an important anti-inflammatory effect, and this effect is linked to its immunomodulatory characteristic. Since the M1 markers were reduced at the same time, M2 markers were increased by the treatment of the macrophages with diene 1.

Synthesis of cinnamic acid ester derivatives with antiproliferative and antimetastatic activities on murine melanoma cells.

Melanoma is the most aggressive skin cancer, and its incidence has continued to rise during the past decades. Conventional treatments present severe side effects in cancer patients, and melanoma can be refractory to commonly used anticancer drugs, which justify the efforts to find new potential anti-melanoma drugs. An alternative to promote the discovery of new pharmacological substances would be modifying chemical groups from a bioactive compound. Here we describe the synthesis of seventeen compounds derived from cinnamic acid and their bioactivity evaluation against melanoma cells. The compound phenyl 2,3-dibromo-3-phenylpropanoate (3q) was the most effective against murine B16-F10 cells, as observed in cytotoxicity and cell migration assays. Simultaneously, this compound showed low cytotoxic activity on non-tumor cells. At the highest concentration, the compound 3q was able to trigger apoptosis, whereas, at lower concentrations, it affected the cell cycle and melanoma cell proliferation. Furthermore, cinnamate 3q impaired cell invasion, adhesion, colonization, and actin polymerization. In conclusion, these results highlight the antiproliferative and antimetastatic potential of cinnamic acid derivatives on melanoma.

Efeito antimicrobiano e anti-inflamatório de ácidos fenólicos isolados, combinados e incorporados em hidrogéis de quitosana para aplicação endodôntica / Antimicrobial and anti-inflammatory effect of phenolic acids, alone, combined and incorporated in chitosan hydrogels for endodontic application

O objetivo do tratamento endodôntico é manter a integridade da raiz, bem como, prevenir ou resolver doenças periapicais, pela erradicação dos microrganismos e de suas fontes de nutrientes provenientes do sistema de canais radiculares. A complexidade da anatomia dos canais radiculares e dos biofilmes multiespécies aumenta a dificuldade em eliminar os microrganismos e controlar a inflamação por procedimentos químico-mecânicos convencionais, o que justifica o uso de medicações intracanais. Novos compostos com amplo efeito antimicrobiano e potencial antiinflamatório, como os ácidos fenólicos, poderiam ser explorados como princípios ativos de medicamentos intracanais. Entretanto, para aumentar a solubilidade, controlar a liberação e estender os efeitos biológicos dos ácidos fenólicos, seria interessante incorporá-los em carreadores de medicamentos como os hidrogéis de quitosana. Este estudo foi dividido em dois capítulos que apresentaram como objetivos: 1) avaliar as atividades antimicrobiana, antibiofilme e antiinflamatória e a citotoxicidade do ácido cinâmico e seus derivados; 2) sintetizar e caracterizar as propriedades químicas e físico-mecânicas de hidrogéis termossensíveis de quitosana e poloxamer contendo ácidos fenólicos, e avaliar o efeito desses hidrogéis sobre biofilmes multiespécies e na viabilidade de macrófagos e fibroblastos. No capítulo 1, a atividade antimicrobiana do ácido cinâmico (CI) e seus derivados ácido cumárico (CO), ácido cafeico (CA), ácido ferúlico (FE) e ácido sinápico (SI) foi avaliada pela determinação da concentração inibitória e bactericida mínima (CIM/CBM) e Concentração Inibitória Fracionada (CIF) sobre Enterococcus faecalis, Streptococcus mutans, Lactobacillus casei, Actinomyces israelii e Fusobacterium nucleatum. Os ácidos fenólicos foram selecionados e seu efeito em biofilmes dual-espécies e multiespécies com as mesmas cepas padrão ou cepas clínicas foram avaliados por contagem bacteriana, microscopia eletrônica de varredura e microscopia confocal. A viabilidade de fibroblastos L929 e macrófagos RAW 264.7 na presença desses ácidos fenólicos foi avaliada por ensaios de resazurina. Além disso, os níveis de mRNA dos marcadores pró-inflamatórios TNF-α, IL-1ß, iNOS e COX-2 foram determinados por PCR quantitativo TaqMan após exposição de macrófagos aos ácidos fenólicos e ao lipopolissacarídeo (LPS). No capítulo 2, foi realizada a síntese e caracterização físicomecânica de hidrogéis de quitosana-poloxamer (CPH) contendo ácidos fenólicos e avaliado seus efeitos no biofilme multiespécies e na viabilidade de macrófagos e fibroblastos. Os dados foram analisados estatisticamente considerando p< 0,05. O ácido cinâmico e o ácido cafeico apresentaram efeito inibitório e bactericida contra todas as espécies bacterianas testadas, com os menores valores de CIM e CBM. Entretanto, não houve efeito sinérgico entre eles (FICI> 0,5). Ambos os compostos (5x a CIM mais alta) foram eficazes na eliminação de biofilmes dual-espécies e na redução significativa de biofilmes multiespécies, especialmente o ácido cinâmico. O ácido cinâmico causou toxicidade mínima para ambas as culturas celulares nas concentrações de CIM e o ácido cafeico não foi citotóxico em concentrações abaixo de 0,125 mg/mL. Ambos os compostos reduziram significativamente TNF-α, IL-1ß, iNOS e COX-2, de maneira dose-dependente. Os CPH foram caracterizados como termorreversíveis e com propriedades mecânicas e bioadesivas desejáveis. O efeito dos hidrogéis CPH+CA (77,8%) e CPH+CI (73,2%) em reduzir os biofilmes multiespécies foi superior ao CPH+ hidroxido de cálcio (CH) (53,6%) e CPH+ clorexidina (CHX) (39,9%). Em geral, CPH + CI causou menor citotoxicidade quando comparado a CPH + CA, para ambas as linhagens celulares. Conclui-se que o ácido cinâmico e ácido cafeico apresentaram efeito bactericida e contra biofilmes formados por bactérias associadas com infecções endodônticas, causando baixa citotoxicidade. Ambos os compostos apresentaram efeito antiinflamatório, inibindo a expressão de marcadores próinflamatórios em macrófagos estimulados por LPS. Os hidrogéis de quitosana-poloxamer foram termorreversíveis e apresentaram adequadas propriedades mecânicas e adesivas para aplicação clínica, e quando combinados principalmente com ácido cinâmico, promoveram a redução de biofilmes multiespécies formados nos túbulos dentinários, causando baixa toxicidade em fibroblastos e macrófagos(AU)

The objective of endodontic treatment is to maintain the integrity of the root, as well as to prevent or resolve periapical diseases, by eradicating microorganisms and their sources of nutrients from the root canal system. The complexity of root canal anatomy and multispecies biofilms increases the difficulty in eliminating microorganisms and controlling inflammation by conventional chemical-mechanical procedures, which justifies the use of intracanal medications. New compounds with broad antimicrobial effect and anti-inflammatory potential, such as phenolic acids, could be explored as active principles of intracanal medications. However, to increase the solubility, control the release and extend the biological effects of phenolic acids, it would be interesting to incorporate them into drug carriers such as chitosan hydrogels. This study was divided into two chapters with the following objectives: 1) to evaluate the antimicrobial, antibiofilm and anti-inflammatory activities and the cytotoxicity of cinnamic acid and its derivatives; 2) synthesize and characterize chemical and physicomecanical properties of thermosensitive chitosan and poloxamer hydrogels containing phenolic acids and evaluate the effect of these hydrogels on multispecies biofilms and on the viability of macrophages and fibroblasts. In chapter 1, the antimicrobial activity of cinnamic acid (CI) and its derivatives coumaric acid (CO), caffeic acid (CA), ferulic acid (FE) and sinapic acid (SI) was evaluated by determining the minimum inhibitory and bactericidal concentration (MIC/MBC) and Fractional Inhibitory Concentration (FIC) on Enterococcus faecalis, Streptococcus mutans, Lactobacillus casei, Actinomyces israelii and Fusobacterium nucleatum. Phenolic acids were selected and their effect on bispecies and multispecies biofilms with the same standard or clinical strains were evaluated by bacterial counts, scanning electron microscopy and confocal microscopy. The viability of L929 fibroblasts and RAW 264.7 macrophages in the presence of these phenolic acids was evaluated by resazurin assays. In addition, mRNA levels of the proinflammatory markers TNF-α, IL-1ß, iNOS and COX-2 were determined by quantitative TaqMan PCR after macrophage exposure to phenolic acids and lipopolysaccharide (LPS). In chapter 2, the synthesis and physical-mechanical characterization of chitosanpoloxamer (CPH) hydrogels containing phenolic acids were performed and their effects on multispecies biofilm and on the viability of macrophages and fibroblasts were evaluated. Data were statistically analyzed considering p< 0.05. Cinnamic acid and caffeic acid showed an inhibitory and bactericidal effect against all bacterial species tested, with the lowest MIC and MBC values. However, no synergistic effect was observed between the compounds (FICI> 0.5). Both compounds (at 5x the highest MIC) were effective in eliminating dual-species biofilms and significantly decreasing multispecies biofilms, especially cinnamic acid. Cinnamic acid caused minimal toxicity to both cell cultures at MIC concentrations and caffeic acid was not cytotoxic at concentrations below 0.125 mg/mL. Both compounds significantly reduced TNF-α, IL1ß, iNOS and COX-2, in a dose-dependent manner. CPH were characterized as thermoreversible and with adequate mechanical and bioadhesive properties. The effect of CPH+CA (77.8%) and CPH+CI (73.2%) hydrogels against multispecies biofilms was superior to CPH + calcium hydroxide (CH) (53.6%) and CPH + chlorhexidine (CHX) (39.9%). In general, CPH + CI caused less cytotoxicity when compared to CPH + CA, for both cell lines. In conclusion, cinnamic acid and caffeic acid showed bactericidal effect and against biofilms of bacteria associated with endodontic infections, causing minimal cytotoxicity. In addition, both compounds showed an anti-inflammatory effect, inhibiting the expression of pro-inflammatory markers in LPS-stimulated macrophages. The chitosan-poloxamer hydrogels were thermoreversible and presented adequate mechanical and bioadhesive properties for clinical application, and when combined specially with cinnamic acid, they promoted the reduction of multispecies biofilms formed in the dentinal tubules, causing low toxicity to fibroblasts and macrophages(AU)