Background: The ongoing treatment modalities for breast cancer (BC) primarily rely on the expression status of ER, PR and HER-2 receptors in BC tissues. Our strategy of chemosensitization provides new insights to counter chemoresistance, a major obstacle that limits the benefits of chemotherapy of mammary cancers.Methods: By utilizing a murine breast cancer model employing NSG mice bearing orthotopic triple-negative breast cancer (TNBC) xenografts, we have evaluated the ability of phytochemical curcumin in chemosensitizing BC to 5-Fluorouracil (5-FU) chemotherapy and the differential modulations of cellular events in response to this strategy, independent of their receptor status.Results: A significant synergistic antitumor potential was observed in the murine model with a sub-optimal dose treatment of 5-FU plus curcumin, as evaluated by a reduction in the tumor-related parameters. We authenticated the pivotal role of thymidylate synthase (TS) in regulating the 5-FU–curcumin synergism using the TNBC pre-clinical model. Our study also confirmed the pharmacological safety of this chemotherapeutic plus phytoactive combination using acute and chronic toxicity studies in Swiss albino mice. Subsequently, the molecular docking analysis of curcumin binding to TS demonstrated the affinity of curcumin towards the cofactor-binding site of TS, rather than the substrate-binding site, where 5-FU binds. Our concomitant in vivo and in silico evidence substantiates the superior therapeutic index of this combination.Conclusion: This is the first-ever pre-clinical study portraying TS as the critical target of combinatorial therapy for mammary carcinomas and therefore we recommend its clinical validation, especially in TNBC patients, who currently have limited therapeutic options.

We previously reported the remarkable potency of uttroside B (Utt-B), saponin-isolated and characterized in our lab from Solanum nigrum Linn, against HCC. Recently, the U.S. FDA approved Utt-B as an ‘orphan drug’ against HCC. The current study validates the superior anti-HCC efficacy of Utt-B over sorafenib, the first-line treatment option against HCC. The therapeutic efficacies of Utt-B vs. sorafenib against HCC were compared in vitro, using various liver cancer cell lines and in vivo, utilizing NOD.CB17-Prkdcscid/J mice bearing human HCC xenografts. Our data indicate that Utt-B holds an augmented anti-HCC efficacy over sorafenib. Our previous report demonstrated the pharmacological safety of Utt-B in Chang Liver, the normal immortalized hepatocytes, and in the acute and chronic toxicity murine models even at elevated Utt-B concentrations. Here, we show that higher concentrations of sorafenib induce severe toxicity, in Chang Liver, as well as in acute and chronic in vivo models, indicating that, apart from the superior therapeutic benefit over sorafenib, Utt-B is a pharmacologically safer molecule, and the drug-induced undesirable effects can, thus, be substantially alleviated in the context of HCC chemotherapy. Clinical studies in HCC patients utilizing Utt-B, is a contiguous key step to promote this drug to the clinic.

Our previous study has demonstrated that Uttroside B (Utt-B), a saponin isolated from the leaves of Solanum nigrum Linn induces apoptosis in hepatic cancer cells and exhibits a remarkable growth inhibition of Hepatocellular Carcinoma (HCC). Our innovation has been granted a patent from the US (US 2019/0160088A1), Canada (3,026,426.), Japan (JP2019520425) and South Korea (KR1020190008323) and the technology have been transferred commercially to Q Biomed, a leading US-based Biotech company. Recently, the compound received approval as ‘Orphan Drug’ against HCC from US FDA, which reveals the clinical relevance of evaluating its antitumor efficacy against HCC. In the present study, we report that Utt-B promotes pro-survival autophagy in hepatic cancer cells as evidenced by the increased expression of autophagy-related proteins, including LC3-II, Beclin1, ATG 5, and ATG 7, as well as a rise in the autophagic flux. Hence, we investigated whether Utt-B-induced autophagic response is complementing or contradicting its apoptotic program in HCC. Inhibition of autophagy using the pharmacological inhibitors, Bafilomycin A1(Baf A1), and 3-methyl adenine (3-MA), and the biological inhibitor, Beclin1 siRNA, significantly enhances the apoptosis of hepatic cancer cells and hence the cytotoxicity induced by Utt-B. We also found increased expression of autophagy markers in Utt-B-treated xenografts derived from HCC. We further analyzed whether the antimalarial drug, Chloroquine (Cqn), a well-known autophagy inhibitor, can enhance the anticancer effect of Utt-B against HCC. We found that inhibition of autophagy using Cqn significantly enhances the antitumor efficacy of Utt-B in vitro and in vivo, in NOD SCID mice bearing HCC xenografts. Taken together, our results suggest that the antitumor effect of Utt-B against HCC can be further enhanced by blocking autophagy. Furthermore, Utt-B in combination with Cqn, a clinically approved drug, if repurposed and used in a combinatorial regimen with Utt-B, can further improve the therapeutic efficacy of Utt-B against HCC.

Despite the advancement in HPV prevention strategies, cervical cancer is a leading cause of cancer death in women worldwide. The anticancer potential of kaempferide, which was derived from Chromolaena odorata, was previously revealed in our in-vitro study. The current investigation aims to confirm the therapeutic efficacy of the molecule against cervical cancer, in-vitro and in-vivo. In NOD-SCID mice with HeLa Luc+ xenografts, kaempferide significantly increases ROS production resulting in a drastic decrease in the luc activity and growth of cervical tumours. It also exhibits down-regulation of oncoproteins E6, E7 and MDM2 and concurrent up-regulation of p53, p21 and pRb. The degradation of phospho-pRb, along with a strong expression of cleaved PARP, TUNEL positivity and a significant decrease in PCNA expression, confirms apoptotic mode of cell death in the treated tissues. Taken together, our study reveals the antioncogenic potential of kaempferide, which regulates oncoproteins and tumor suppressors accordingly. This is the first report depicting kaempferide as an inhibitor of HPV oncoproteins and hence as a candidate drug molecule against cervical cancer.

Metabolic dysfunction-associated steatohepatitis (MASH) is characterized by excessive accumulation of fat, accompanied by inflammation and liver injury, ultimately triggering chronic conditions, including fibrosis and cirrhosis, which may progress to hepatocellular carcinoma (HCC). Uttroside B (Utt-B), a phytosaponin isolated in our lab, has gained global recognition owing to its anti-HCC potential and is currently a United States FDA-designated ‘orphan drug’ against HCC. The present study highlights Utt-B as a propitious candidate drug against MASH and MASH-induced HCC. MASH and MASH-induced HCC were developed in C57BL/6J mice using two distinct murine models: a high-fat diet model and a streptozotocin-induced steatohepatitis-derived HCC model, followed by i.p. administration of Utt-B. Protein expression analysis was performed using real-time quantitative reverse transcription-PCR and immunoblotting, while H&E, Oil Red O, Sirius Red, and Masson’s Trichrome were utilized for staining cells/tissues. Nanostring n-Counter analysis was used to investigate the mechanism underlying the antifibrotic effects of Utt-B in MASH-induced HCC. Proliferation and apoptosis markers were also evaluated. Statistical analyses were conducted using R and GraphPad-Prism with significance set at p <0.05. Utt-B ameliorated MASH-associated pathological features, including steatosis, hepatocyte ballooning, and inflammation (N = 6; non-alcoholic fatty liver disease activity score [NAS] <2, p <0.0001). Utt-B upregulated the expression of autophagy markers autophagy-related 7 (ATG-7), Beclin-1, and microtubule-associated protein 1A/1B-light chain (LC3-II), and downregulated the expression of α-smooth muscle actin (α-SMA), which indicates the activation of hepatic stellate cells. Utt-B also halted the progression of MASH to HCC by hindering development of fibrosis with simultaneous inhibition of proliferative signals and induction of apoptosis in murine models (N = 6; NAS <3, p <0.01). Our investigation revealed effective impedance of MASH and its concomitant progression to HCC by Utt-B. Given the lack of anti-MASH drugs, these findings establish Utt-B as a potent drug for treating MASH and MASH-induced HCC. MASH, stage IV of non-alcoholic fatty liver disease, can lead to chronic conditions, including fibrosis and cirrhosis, elevating the risk of HCC. Impaired lipid metabolism, inflammation, inhibition of autophagy, and dysregulated collagen and extracellular matrix biosynthesis are key factors contributing to the advance of MASH to HCC. Our discovery of Utt-B, a phytosaponin that exhibits remarkable anti-HCC potential and is a United States FDA-designated orphan drug against HCC, has gained global recognition. The present study reveals Utt-B as a propitious candidate drug against MASH and MASH-induced HCC in a high-fat diet murine model and streptozotocin-induced steatohepatitis-derived HCC animal model, respectively.