Glioblastoma (GBM) is a fatal cancer with a dismal prognosis and a dire need for novel chemotherapeutics. Metabolic reprogramming is an established hallmark of cancer. In our previous study on GBM, we aimed at targeting the metabolic reprogramming of cancer by using stiripentol (STP), a putative lactate dehydrogenase (LDH) inhibitor and an FDA-approved anti-epileptic drug. However, the precise mechanism of STP’s anti-cancer activity remains unclear. We aimed to elucidate the mechanism of action of STP in GBM to further develop STP as a therapeutic. We employed a multiomic approach followed by metabolic and cellular assays. STP treatment induced genetic and metabolic alterations in GBM cells. Inhibition of LDH by STP was moderate but not potent. The cellular changes were accompanied by an increase in reactive oxygen species, a decrease in mitochondrial membrane potential, and induction of senescence in GBM cells. Our research indicates that further research in senescence-inducing agents and novel LDH inhibitors can provide novel therapeutics for GBM.

The administration of traditional chemotherapeutic drugs to treat lung cancer has obvious disadvantages such as toxic side effects, the development of drug resistance, the need for combinational therapy, and poor bioavailability. To address the limitations of drugs and improve therapeutic efficacy, we developed novel transferrin ligand-conjugated resveratrol-cyclodextrin-loaded nanoparticles (CD-RES TNPs), which are capable of actively targeting the transferrin receptor (TfR), overexpressed on non-small cell lung cancer (NSCLC) cells. The results demonstrated that the % encapsulation efficiency (%EE) was approximately 90%. Cytotoxicity studies in the NSCLC cell line H1299 resulted in significant reductions in IC50 values with Tf-conjugated nanoparticles compared to plain RES or non-targeted NPs, along with reduced % colony growth, cell migration, and considerable apoptosis induction, while also retaining the antioxidant activity of the phytochemical. Hemolysis assay suggested that the TNPs exhibited good biosafety and were suitable for clinical applications and patient acceptability. Additionally, 3D spheroid studies exhibited enhanced efficiency of conjugated nanoparticles in penetrating the tumor tissue, and inhibiting growth. The current findings helped lay the foundation for using CD-RES TNPs in treating lung cancer.

INTRODUCTION:Endothelins (ETs) are a family of versatile peptides composed of 21 amino acids with three isoforms: ET-1, ET-2, and ET-3. As the most abundant of the three isoforms, ET-1 is involved in various biological processes, such as regulation of vascular tone, humoral homeostasis, and neural crest development. However, focus is now being directed towards investigating the functions of the ET axis in the progression of different tumor types including ovarian, prostate, breast, lungs etc. HJP 272 is a novel ETAR antagonist and while our group has previously researched its effects on lung inflammation and preterm birth, this study marks the first time its role in cancer has been explored. METHODS:We evaluated the in vitro activities of HJP 272 in the ET-1 and ETAR overexpressing cell lines MDA-MB-231 (TNBC), and A549 (NSCLC). While HJP 272 had no effect on the viability of cancer cells, we observed a significant inhibition in the migration, invasion, and clonogenic capacities of both cell lines. RNA-seq and western blot data demonstrate the potential underlying molecular mechanisms of this compound in vitro. Furthermore, HJP 272 was evaluated in a 3D spheroid assay for its ability to inhibit tumor formation in both cell lines, revealing a significant change in MDA-MB-231 cells while no significant changes were observed in A549 cells. CONCLUSIONS:Our work indicates a therapeutic potential for HJP 272 in cancer metastasis. The distinct outcomes between the two cell lines shed light on the potential differences of HJP 272's effects across multiple cancer types.

Gankyrin (PSMD10) is a 25 kDa oncogenic protein and regulatory subunit of the 26S proteasome, characterized by a sevenfold ankyrin repeat domain. Gankyrin is overexpressed in various malignancies, particularly gastrointestinal (GI) cancers. Gankyrin contributes to tumorigenesis by modulating key signaling pathways and engaging in oncogenic protein-protein interactions with tumor suppressors, including p53 and Rb, thereby promoting cell proliferation, metastasis, and resistance to treatment. Recent advances have shed light on the structural basis of gankyrin's molecular interactions, its potential as a diagnostic and prognostic biomarker, and emerging therapeutic strategies. Together, targeting gankyrin represents a promising strategy for precision oncology in GI cancers.

Background: Gankyrin is an ankyrin-repeat protein that promotes cell proliferation, tumor development and cancer progression when overexpressed. Aim: To design and synthesize a novel series of gankyrin-binding small molecules predicated on a 2,5-pyrimidine scaffold. Materials & methods: The synthesized compounds were evaluated for their antiproliferative activity, ability to bind gankyrin and effects on cell cycle progression and the proteasomal degradation pathway. Results: Compounds 188 and 193 demonstrated the most potent antiproliferative activity against MCF7 and A549 cells, respectively. Both compounds also demonstrated the ability to effectively bind gankyrin, disrupt proteasomal degradation and inhibit cell cycle progression. Conclusion: The 2,5-pyrimidine scaffold exhibits a novel and promising strategy for binding gankyrin and inhibiting cancer cell proliferation.

Liver cancer is a complex disease that involves various oncoproteins and the inactivation of tumor suppressor proteins (TSPs). Gankyrin is one such oncoprotein, first identified in human hepatocellular carcinoma, that is known to inactivate multiple TSPs, leading to proliferation and metastasis of tumor cells. Despite this, there has been limited development of small molecule gankyrin binders for the treatment of liver cancer. In this study, we are reporting the structure-based design of gankyrin-binding small molecules which inhibit the proliferation of HuH6 and HepG2 cells while also increasing the levels of certain TSPs, such as Rb and p53. Interestingly the first molecule to exhibit inhibition by 3D structure stabilization is seen. These results suggest a possible mechanism for small-molecule inhibition of gankyrin and demonstrate that gankyrin is a viable therapeutic target for the treatment of liver cancer.

The majority of novel targeted anticancer drugs are weakly basic molecules with poor aqueous solubility which ultimately compromise their oral bioavailability. Lenvatinib (Lnv) is indicated for the first-line treatment of patients with hepatocellular carcinoma (HCC). Since it is a classical brick-dust molecule with extremely poor aqueous, organic solvent and oil solubility, Lnv formulation development is very challenging. We hypothesized that combining Lnv with valproic acid (a histone deacetylase (HDAC) inhibitor) will improve oil solubility and water hydrophobic ion pairing, as well as anticancer activity via synergistic interactions. Self-Nanoemulsifying Preconcentrate (SNEP) of Lnv was developed by screening and optimizing various biocompatible co-solvents, oils and non-ionic surfactants. Lnv and Valproic Acid (VA) loaded SNEP (LnVaNo) were optimized for kinetic stability, particle size, polydispersity index and in vitro release dissolution. In vitro cytotoxicity of LnVaNo was investigated in comparison with the individual drugs using an MTT assay in three hepatic cancer cell lines (Hep3B, HepG2, and HuH6) along with migration and clonogenic assays. The anticancer efficacy was also analyzed in 3D multicellular hepatic tumor spheroids by tumor growth analysis and a Live/Dead cell assay. The interaction of Lnv with VA enhanced the solubilization and improved the kinetic stability of Lnv in nanoglobules. The optimized Lnv-VA system spontaneously formed a homogenous dispersion with a particle size <100 nm. LnVaNo showed a 3-4-fold enhancement in IC50 against in the hepatic cancer cell lines compared to Lnv alone. Significantly higher red fluorescence (necrotic surface) and marked inhibition in tumor spheroid growth compared to the individual drug and control after 10 days of treatment indicated the superior efficacy of LnVaNo in tumor penetration and growth inhibition. Valproic acid facilitates molecular interaction to improve the solubility of weakly basic drugs and anticancer efficacy for the treatment of cancer.

Sorafenib (Soraf) is a second-generation tyrosine kinase inhibitor repurposed against different cancers by working on various molecular pathways, playing a vital role in cancer cell proliferation. Sorafenib is a BCS class II drug, and its low solubility further hinders its bioavailability and results in reduced therapeutic efficacy. Therefore, in this study, we aim to explore the potential of β-cyclodextrins (β-CD) derivatives to form an inclusion complex with sorafenib to enhance sorafenib's solubility, stability, and therapeutic efficacy. Our findings suggest that HP-β-CD portrayed a higher affinity towards sorafenib than SBE-β-CD and significantly improved the solubility of sorafenib. Additionally, spectroscopic analysis and solid-state studies provided valuable information regarding the intermolecular interaction between sorafenib and HP-β-CD.Furthermore, the stability of sorafenib in biological fluids and under physiological conditions was maintained under the influence of the inclusion complex, whereas plain sorafenib degraded quickly. In addition, β-CD complexation significantly improved sorafenib's intestinal permeability, as shown in physiologically relevant EpiIntestinal® models. Lastly, the sorafenib inclusion complex demonstrated increased cytotoxicity in NSCLC and HCC cell lines. This study shows the potential of HP-β-CD to improve the solubility and therapeutic efficacy of sorafenib, thus extending its clinical application.

Tuberculosis (TB) is a contiguous airborne disease caused by Mycobacterium tuberculosis ( M.tb ), primarily affecting the human lungs. The progression of drug-susceptible TB to drug-resistant strains, MDR-TB and XDR-TB, has become a global challenge toward eradicating TB. Conventional TB treatment involves frequent dosing and prolonged treatment regimens predominantly by an oral or invasive route, leading to treatment-related systemic adverse effects and patient’s noncompliance. Pulmonary delivery is an attractive option as we could reduce dose, limit systemic side-effects, and achieve rapid onset of action. Delamanid (DLD), an antituberculosis drug, has poor aqueous solubility, and in this study, we aim to improve its solubility using cyclodextrin complexation. We screened different cyclodextrins and found that HP-β-CD resulted in a 54-fold increase in solubility compared to a 27-fold and 13-fold increase by SBE-β-CD and HP-ɣ-CD, respectively. The stability constant (265 ± 15 M^−1) and complexation efficiency (8.5 × 10^−4) suggest the formation of a stable inclusion complex of DLD and HP-β-CD in a 2:1 ratio. Solid-state characterization studies (DSC, PXRD, and NMR) further confirmed successful complexation of DLD in HP-β-CD. The nebulized DLD-CD complex solution showed a mass median aerodynamic diameter of 4.42 ± 0.62 μm and fine particle fraction of 82.28 ± 2.79%, suggesting deposition in the respiratory airways. In bacterial studies, minimum inhibitory concentration of DLD-CD complex was significantly reduced (four-fold) compared to free DLD in M.tb (H37Ra strain). Furthermore, accelerated stability studies confirmed that the inclusion complex was stable for 4 weeks with 90%w/w drug content. In conclusion, we increased the aqueous solubility of DLD through cyclodextrin complexation and improved its efficacy in vitro . Graphical Abstract

Non-small cell lung cancer (NSCLC) is ranked first worldwide amongst deadly cancers with poor patient survival rate. While chemotherapy, radiation and surgery are available treatment options, NSCLC tumor cells are known to develop resistance and quickly metastasize. Recently, PARP inhibitors have shown their potential as anticancer agents. One of the FDA approved PARP inhibitors, Olaparib (Ola) has been approved for advanced and metastatic ovarian, prostate, pancreatic and breast cancers; and has shown potential in NSCLC treatment. However, due to its low solubility and permeability, Ola possesses poor bioavailability leading to minimal therapeutic efficacy. In this study, we aimed to explore the potential of FDA approved pharmaceutical excipient, Polyvinylpyrrolidone (PVP), as a complexation agent to overcome the challenges of Olaparib by developing an inhaled dosage form for NSCLC treatment. Phase solubility studies indicated enhanced solubility of Ola when complexed with PVP. Complex formation between Ola and PVP was confirmed via solid state characterization studies. Accelerated stability study showed that Olaparib was not degraded at 37 degrees C when complexed with PVP, suggesting PVP rendering stability to Olaparib, thus shielding it from harsh environment. The formed Ola-PVP complex demonstrated excellent aerosolization performance and exhibited a higher cytotoxic profile against NSCLC cell lines. 3D tumor simulation studies also depicted improved therapeutic potential of Ola against an actual physiological tumor. This study lays the foundation for preclinical and potential clinical translation of inhaled Olaparib with PVP.

Malignant pleural mesothelioma (MPM) is a rare and aggressive cancer affecting the pleural lining of the lungs. Celastrol (Cela), a pentacyclic triterpenoid, has demonstrated promising therapeutic potential as an antioxidant, anti-inflammatory, neuroprotective agent, and anti-cancer agent. In this study, we developed inhaled surface-modified Cela-loaded poly(lactic-co-glycolic) acid (PLGA) microparticles (Cela MPs) for the treatment of MPM using a double emulsion solvent evaporation method. The optimized Cela MPs exhibited high entrapment efficiency (72.8 ± 6.1%) and possessed a wrinkled surface with a mean geometric diameter of ~2 µm and an aerodynamic diameter of 4.5 ± 0.1 µm, suggesting them to be suitable for pulmonary delivery. A subsequent release study showed an initial burst release up to 59.9 ± 2.9%, followed by sustained release. The therapeutic efficacy of Cela MPs was evaluated against four mesothelioma cell lines, where Cela MP exhibited significant reduction in IC50 values, and blank MPs produced no toxicity to normal cells. Additionally, a 3D-spheroid study was performed where a single dose of Cela MP at 1.0 µM significantly inhibited spheroid growth. Cela MP was also able to retain the antioxidant activity of Cela only while mechanistic studies revealed triggered autophagy and an induction of apoptosis. Therefore, these studies highlight the anti-mesothelioma activity of Cela and demonstrate that Cela MPs are a promising inhalable medicine for MPM treatment.

Glioblastoma multiforme (GBM) is a highly proliferative grade IV malignant astrocytoma with a 5-year survival rate of ~5%. The current treatment strategies for GBM including surgery, radiation, and chemotherapy face multiple challenges such as resistance to standard chemotherapeutic temozolomide (TMZ), sensitive location, and presence of blood-brain barrier that hinders permeation of various chemotherapeutics. The dire need and urgency to develop a new therapeutic for the treatment of GBM led us to explore metabolic vulnerabilities of cancer. Stiripentol (STP) is an FDA approved drug used for the treatment of Dravet's syndrome (a rare type of epilepsy) and a putative LDH inhibitor that we screened out of many metabolic inhibitors for GBM. STP showed efficacy in cytotoxicity and proliferation assay on U87 and U138 GBM cells. STP also showed efficacy in clonogenic assay and wound healing assay in GBM cells. To further explore and discover the mechanistic target of STP for the treatment of GBM, we performed apoptosis assay, cell cycle assay, and studied the expression of proteins involved in apoptosis, metastasis, and cell signaling. In addition, 3D spheroid study was performed to mimic the tumor microenvironment and the effect of STP was assessed on the same. Next, synergy studies of STP with standard of care TMZ was performed on U87 cells. Cellular toxicity studies were performed to evaluate the safety profile of STP and the combination treatment with TMZ on normal human cells. We will elucidate mechanistic basis of STP anticancer activity in GBM cells and test preclinical potential of STP in tumor xenograft model. Therefore, STP is an effective compound that can augment TMZ for GBM treatment and merits further investigation.

Background: Gankyrin, a member of the 26S proteasome, is an overexpressed oncoprotein in hepatoblastoma (HBL) and hepatocellular carcinoma (HCC). Cjoc42 was the first small molecule inhibitor of Gankyrin developed; however, the IC50 values of >50 μM made them unattractive for clinical use. Second-generation inhibitors demonstrate a stronger affinity toward Gankyrin and increased cytotoxicity. The aim of this study was to characterize the in vitro effects of three cjoc42 derivatives. Methods: Experiments were performed on the HepG2 (HBL) and Hep3B (pediatric HCC) cell lines. We evaluated the expression of TSPs, cell cycle markers, and stem cell markers by Western blotting and/or real-time quantitative reverse transcription PCR. We also performed apoptotic, synergy, and methylation assays. Results: The treatment with cjoc42 derivatives led to an increase in TSPs and a dose-dependent decrease in the stem cell phenotype in both cell lines. An increase in apoptosis was only seen with AFM-1 and -2 in Hep3B cells. Drug synergy was seen with doxorubicin, and antagonism was seen with cisplatin. In the presence of cjoc42 derivatives, the 20S subunit of the 26S proteasome was more available to transport doxorubicin to the nucleus, leading to synergy. Conclusion: Small-molecule inhibitors for Gankyrin are a promising therapeutic strategy, especially in combination with doxorubicin.

Polypharmacology is a concept where a molecule can interact with two or more targets simultaneously. It offers many advantages as compared to the conventional single-targeting molecules. A multi-targeting drug is much more efficacious due to its cumulative efficacy at all of its individual targets making it much more effective in complex and multifactorial diseases like cancer, where multiple proteins and pathways are involved in the onset and development of the disease. For a molecule to be polypharmacologic in nature, it needs to possess promiscuity which is the ability to interact with multiple targets; and at the same time avoid binding to antitargets which would otherwise result in off-target adverse effects. There are certain structural features and physicochemical properties which when present would help researchers to predict if the designed molecule would possess promiscuity or not. Promiscuity can also be identified via advanced state-of-the-art computational methods. In this review, we also elaborate on the methods by which one can intentionally incorporate promiscuity in their molecules and make them polypharmacologic. The polypharmacology paradigm of “one drug-multiple targets” has numerous applications especially in drug repurposing where an already established drug is redeveloped for a new indication. Though designing a polypharmacological drug is much more difficult than designing a single-targeting drug, with the current technologies and information regarding different diseases and chemical functional groups, it is plausible for researchers to intentionally design a polypharmacological drug and unlock its advantages.