The hypovascular nature of pancreatic tumors creates a nutrient-scarce, hypoxic microenvironment, yet pancreatic cancer cells adapt by altering their metabolism to thrive under austere conditions-a phenomenon known as "austerity." Targeting this adaptation offers a promising strategy for next-generation therapeutics that selectively impair pancreatic cancer cell viability in nutrient-deprived states without toxicity under nutrient-rich conditions. Here, we evaluated the anti-pancreatic cancer properties of grandifloridin D, a synthetic derivative of (+)-grandifloracin. In vitro antiausterity assays demonstrated that grandifloridin D potently and preferentially reduced the viability of MIA PaCa-2 pancreatic cancer cells under nutrient deprivation at a PC concentration of 0.14 μM. Live-cell imaging and ethidium bromide/acridine orange (EB/AO) dual staining confirmed that grandifloridin D induces cell death by disrupting membrane integrity. Under nutrient-rich conditions, grandifloridin D exhibited antimetastatic activity, significantly inhibiting MIA PaCa-2 cell migration in real-time assays and suppressing colony formation and spheroid formation. Western blot analysis revealed that grandifloridin D is a potent inhibitor of the protein kinase B (Akt) and mammalian target of rapamycin (mTOR) signaling pathway while also suppressing the autophagy-related proteins microtubule-associated protein 1 light chain 3 (LC3). These results suggest that grandifloridin D is a promising antiausterity agent for pancreatic cancer drug development.

Radiation therapy exerts its therapeutic effect by killing cells via the induction of DNA double-strand breaks (DSBs) in malignant tumors, but cancer cells can repair damaged DNA, leading to radiation resistance (radioresistance). Therefore, a radiosensitizing effect can be expected by suppressing mechanism(s) involved in DNA repair after irradiation. Here, we show that the P2Y12 receptor is involved in the radioresistance of mouse B16 melanoma cells, and that P2Y12 antagonist treatment decreases the radioresistance both in vitro and in vivo by inhibiting DNA repair after γ-irradiation. P2Y12 receptor antagonists Clopidogrel and PSB0739 increased cellular sites of unrepaired DNA by suppressing the DNA damage response (DDR) after γ-irradiation and enhanced radiation-induced proliferative death in B16 melanoma cells. On the other hand, ADP (a P2Y12 receptor agonist) enhanced DDR after γ-irradiation and increased radioresistance. Knockdown of the P2Y12 receptor resulted in an increase of unrepaired DNA damage and enhanced proliferative death after γ-irradiation. Suppression of the P2Y12 receptor also contributed to the enhancement of the cancer-killing effect of γ-irradiation, even in fractionated irradiation samples in which the cancer-killing effect decreased due to sublethal damage recovery. Finally, PSB0739 significantly enhanced the antitumor effect of γ-irradiation in vivo. Our results suggest that P2Y12 receptor antagonists are promising candidates as radiosensitizers to improve radiation therapy.

Opioid analgesics are indispensable therapeutic agents for patients experiencing cancer pain. Recently, immune checkpoint inhibitors (ICIs) have made substantial progress in cancer treatment, and the range of cancer types for which they are applicable has expanded. Consequently, the combined use of ICIs with other drugs has also increased. However, it has been pointed out that ICIs may interfere with opioid nociception and weaken the analgesic effects of opioids. Therefore, in this study, we investigated the drug-drug interactions between opioid analgesics and ICIs in patients with cancer, especially their influence on analgesic effects. A multicenter collaborative study was conducted to investigate changes in pain intensity in patients with cancer pain who were prescribed opioid analgesics and ICIs. As a result, when opioids were administered to patients receiving ICIs, the median pain intensity, measured using the Numerical Rating Scale (NRS), significantly decreased from 4.5 at baseline to 3.0 on Day 1 and 2.5 on Day 7, demonstrating favorable analgesic effects. Similarly, when ICIs were administered to patients receiving opioids, the median pain intensity (NRS) significantly decreased from 3.0 at baseline to 2.0 on Day 1 and 2.75 on Day 7. These results suggest that opioids provided effective analgesia when administered alongside ICIs.

Metabolic enzymes are occasionally downregulated in in vitro induction studies. Recently, HepaRG cells have been used for CYP induction assays instead of human hepatocytes in the early drug discovery stage; however, there is limited information on CYP downregulation by drug stimulation. In this study, we evaluated the effect of hypoxia-inducible factor-prolyl hydroxylase (HIF-PH) inhibitors, which downregulate CYP in human hepatocytes, on CYP gene expression in HepaRG cells. Microarray analysis to determine the expression levels of pharmacokinetics-related enzymes and RT-PCR to determine the expression levels of CYP3A4, CYP2B6, CYP1A2, and their nuclear receptor mRNA were conducted in HepaRG cells treated with HIF-PH inhibitors. Treatment of HepaRG cells with HIF-PH inhibitors decreased the expression of several pharmacokinetics-related metabolic enzymes, whereas Erythropoietin (EPO) and Pyruvate Dehydrogenase Kinase1 (PDK1) genes were induced. The expression of CYP3A4 and CYP2B6 in HepaRG cells showed concentration- and time-dependent downregulation following treatment with the HIF-PH inhibitor. The downregulation of these enzymes was correlated with the decrease of PXR/RXRα and CAR/RXRα, respectively. CYP1A2 decreased transiently, but recovered with continued HIF-PH inhibitor treatment. CYP3A4 and CYP2B6 were downregulated by HIF-PH inhibitors in HepaRG cells and human hepatocytes. In contrast, CYP1A2 in HepaRG cells responded differently to HIF-PH inhibitors than in human hepatocytes. Since CYP downregulation is commonly observed with HIF-PH inhibitors, along with the induction of EPO and PDK1 genes, stabilizing HIF may be one of the factors involved in CYP downregulation.

Tumor necrosis factor-α inhibitors (TNFis) are associated with a risk of paradoxical psoriasis, but quantitative data remain limited. One proposed mechanism is the induction of interferon (IFN) production following TNFi administration. Etanercept and certolizumab pegol, which contain immunoglobulin fragments in their structures, reportedly induce IFN production in T cells more than monoclonal antibody (mAb) TNFi agents. Based on this, we hypothesized that non-mAb TNFi agents might carry a higher risk of paradoxical psoriasis than mAb agents. This study compared the risk of paradoxical psoriasis between mAb and non-mAb TNFi agents in rheumatoid arthritis (RA) patients. Using a claims database, we identified 1577 subjects in the mAb group and 1517 in the non-mAb group. Patient characteristics, including sex, age, and prior RA treatment, were extracted, and the onset of psoriasis was identified. Multivariable Cox regression analysis showed the hazard ratio (HR) for psoriasis onset in the mAb group versus the non-mAb group was 1.66 (95% confidence interval [CI]: 0.79-3.48). Subgroup analyses revealed that compared to etanercept, the HR for adalimumab was 1.43 (95% CI: 0.49-4.19), and compared to certolizumab pegol, it was 0.67 (95% CI: 0.19-2.39). These findings suggest that our hypothesis was not supported and that the risk of paradoxical psoriasis may vary even among non-mAb agents, as indicated by differences observed between etanercept and certolizumab pegol.

Retinoic acid receptor-related orphan receptor gamma (RORγ) is a key transcriptional factor that plays a crucial role in the differentiation and activation of Type 17 cells, such as interleukin-17 (IL-17)-producing CD4 T (Th17) cells and CD8 T (Tc17) cells, which are known to boost antitumor responses. Although a RORγ agonist (LYC-55716) has been under clinical evaluation, the precise effects of RORγ agonists on immune cells within tumor environments remain unclear. In our study, we investigated the role of tumor-infiltrating immune cells in the MC38 syngeneic mouse model of colorectal cancer using Compound-34, a novel orally available RORγ-selective agonist we discovered. Our findings revealed that Compound-34 exerts its antitumor efficacy by modulating immune cell activity rather than directly targeting tumor cells. Specifically, Compound-34 increased the infiltration of effector T cells, including Th17 and Tc1 (interferon [IFN]-γ CD8 T) cells, as well as innate immune cells like natural killer (NK) and natural killer T (NKT) cells, within the MC38 tumor tissue. Following the administration of Compound-34, there was an increase in IFNγ and Granzyme B within the MC38 tumor tissue, accompanied by an increase in the infiltration of cytotoxic immune cells. Moreover, the addition of Th17-derived cytokines to MC38 cells stimulated the release of CXCL10, a chemokine crucial for immune cell recruitment. These results offer valuable insights into the immunomodulatory and therapeutic potential of RORγ agonists in cancer immunotherapy, highlighting their role in enhancing immune cell infiltration and activity within tumors.

Glioblastoma (GBM) is the most malignant form of glioma. Glioma stem cells (GSCs) contribute to the initiation, progression, and recurrence of GBM via their self-renewal potential and tumorigenicity. This review presents a brief overview of the influences of two protein posttranslational modifications in GSCs on the properties of GSCs and malignancy of GBM: (I) the ubiquitination of transforming growth factor (TGF)-β receptor (TGFBR) by SMAD-specific E3 ubiquitin protein ligase 2 (SMURF2) and (II) the phosphorylation of extracellular signal-regulated kinase 5 (ERK5) by mitogen-activated protein kinase/ERK kinase 5 (MEK5). The phosphorylation state of SMURF2Thr249 regulates the stemness and tumorigenicity of GSCs via the ubiquitin-dependent modification of the TGFBR-SMAD-SOX axis, along with the downregulation of SMURF2Thr249 phosphorylation in patients with GBM. MEK5 controls the self-renewal and tumorigenic potential of GSCs by phosphorylating the ERK5-signal transducer and activator of transcription 3 (STAT3) axis concomitant with high expression and activity of ERK5 in GSCs. These findings contributed to our understanding of the molecular mechanisms underlying the maintenance of the stemness and tumorigenicity of GSCs through protein posttranslational modifications. We propose that these two protein posttranslational modifications in GSCs might be explored as an effective therapeutic approach against various cancers whose malignancies are associated with the stemness of cancer stem cells.

In Japan, the simple N-substituted substances triethylamine and trimethylamine have been designated possibly hazardous air pollutants also requiring further investigation to protect the aquatic environment. Triethylamine is relevant to human biomonitoring in manufacturing workers because of its possible adverse effects; in contrast, trimethylamine exposure from normal daily dietary consumption is considered nontoxic. Although a role for flavin-containing monooxygenase 3 in the metabolism of triethylamine was recently reported, no simplified physiologically based pharmacokinetic (PBPK) model to estimate human plasma and urinary levels of triethylamine and its N-oxide has currently been developed. In this study, in silico human plasma and urine concentrations of triethylamine were estimated after virtual oral administration using a newly established triethylamine PBPK model. The results were compared with our previously established trimethylamine PBPK model. In silico plasma and urinary concentration curves were generated after single virtual administrations of triethylamine and trimethylamine. After 28 d of daily exposure to reported maximum oral doses of triethylamine and trimethylamine from the public water supply of 0.02 and 0.68 µg/kg body weight/d, respectively, the mean modeled urinary levels for the final day were 3.7 and 1.1 pmol/mL. The proposed occupational standard of 10 mg triethylamine/m of air reportedly corresponds to a urinary excretion of approx. 0.4 nmol/mL. The results of the current forward dosimetry analyses, therefore, indicate at least a two-order safety margin (drinking water versus occupational standard) for triethylamine. The present PBPK model for triethylamine and its N-oxide could estimate daily exposures using forward dosimetry and thereby facilitate risk assessment in humans.

Exposure to ionizing radiation during early pregnancy poses serious risks to fetal development, particularly during organogenesis. In this study, we evaluated the protective effects of AHCC, a standardized extract of Lentinula edodes mycelia with known immunomodulatory and metabolic properties, using an ICR mouse model. Intraperitoneal administration of AHCC prior to 1.4-Gy γ-irradiation on gestational day 9 significantly reduced the incidence of tail malformations and diaphragmatic hernias, without affecting implantation or fetal survival. Additionally, lifelong AHCC ingestion across three generations did not alter reproductive parameters, namely, gestation length and litter size, supporting its safety during pregnancy. These findings suggest that AHCC may mitigate radiation-induced teratogenic effects through maternal immune modulation, without compromising reproductive outcomes.

Erectile responses in reproduction have not been analyzed extensively because of the limitation of their visualization analyses of inner penile structure and its dynamic changes. In addition, the complex and rapid changes of erectile responses require new manipulation techniques suitable to regulate erected and flaccid status. The current review describes novel strategies for the above visualization of erection and regulation of the erectile responses.

This study established experimental models of acute lung injury (ALI) both in vitro, using human alveolar epithelial cells, and in vivo, using murine models, with ALI induced by lipopolysaccharide (LPS). Propofol was administered to the animal models, and its effects on alveolar epithelial cell damage, lung tissue pathology, and inflammatory mediator levels were evaluated to assess its potential protective role against lung injury. The results showed that LPS-induced reactive oxygen species (ROS) played a central role in the development of ALI in both the cell and animal models, triggering pyroptosis of alveolar epithelial cells. This, in turn, led to the release of pro-inflammatory cytokines, exacerbating the lung injury. Importantly, propofol was found to reduce LPS-induced ALI by inhibiting ROS production and modulating key proteins involved in the pyroptosis pathway. These findings offer new insights into the mechanisms underlying ALI, providing a basis for the development of clinical treatments and suggesting promising therapeutic strategies for ALI patients.

Dai-kenchu-to is used to prevent and relieve ileus post-gastrectomy; however, it has been reported to cause hepatobiliary injury in 0.3% of patients, and knowledge regarding its risk in this context is limited. In this study, we aimed to evaluate the effect of dai-kenchu-to, a representative traditional Japanese herbal medicine, on the risk of hepatobiliary injury in patients with gastric cancer who underwent gastrectomy, using real-world data from acute care hospitals. The diagnosis procedure combination data of patients admitted with gastric cancer who underwent gastrectomy between April 2008 and May 2019 were analyzed. The collected data included sex, age, comorbid injuries and illnesses at admission, medication records during hospitalization, and tumor node metastasis classifications. The primary outcome, occurrence of hepatobiliary injury, was identified using records of the use of hepatoprotective agents or cholagogues. The odds ratios of dai-kenchu-to for predisposition to hepatobiliary injury after gastrectomy were estimated using multivariate logistic regression analysis. Herein, 33137 gastric cancer patients who underwent gastrectomy were included in the analysis. The incidences of hepatobiliary injury among patients in the dai-kenchu-to-exposed and non-exposed groups were both 5.9%. The multivariate odds ratio of dai-kenchu-to exposure for hepatobiliary injury was 0.97 (95% confidence interval, 0.73-1.29; p = 0.847). In this study, no risk of moderate-to-severe hepatobiliary injury due to the postoperative administration of dai-kenchu-to was noted in patients with gastric cancer who had undergone gastrectomy. These results provide information regarding the safety of dai-kenchu-to use after gastrectomy in patients with gastric cancer.

Histone deacetylases (HDACs) regulate chromatin structure and gene expression, and their inhibition has been proposed as a radioprotective strategy. However, few studies have compared multiple HDAC inhibitors (HDACis) under identical conditions. This study evaluated the efficacy of seven HDACis in a mouse total body irradiation (TBI) model. Male ICR mice received 7.5 Gy TBI followed by a single administration of valproic acid (VPA; 300 or 600 mg/kg), sodium butyrate (NaB; 500 or 1000 mg/kg), trichostatin A (TSA; 0.5 or 1.0 mg/kg), vorinostat (10 or 50 mg/kg), panobinostat (25 or 50 mg/kg), givinostat (5 or 10 mg/kg), or entinostat (25 or 50 mg/kg). Survival was monitored for 20 d. Only VPA at 600 mg/kg significantly improved survival compared with vehicle (overall p = 0.00241; p = 0.0039 vs. vehicle), while all other HDACis showed no significant benefit. VPA's efficacy may reflect a combination of effects on DNA repair, inflammation, and redox regulation rather than HDAC inhibition alone. These findings suggest VPA to be a promising candidate for radioprotection and emphasize the need for further studies to optimize dosing and explore underlying mechanisms.

The aim of this study was to investigate whether ginsenoside Rb1 attenuates cockroach extract (CRE)-induced asthma by interfering with mitochondrial dysfunction. After induction with CRE, mice were administered different doses of Rb1. Hematoxylin-eosin (H&E) staining, enzyme-linked immunosorbent assay (ELISA), and flow cytometry analysis revealed that inflammatory cell infiltration, total immunoglobulin E (IgE) and CRE-specific IgE in serum, and inflammatory cytokines in bronchoalveolar lavage fluid were effectively inhibited by Rb1. Through Western blot, TUNEL, and immunofluorescence colocalization assays, we observed Rb1 also inhibited endogenous reactive oxygen species (ROS), tightly associated with increased superoxide dismutase, catalase levels, and decreased malondialdehyde levels. Subsequently, the silent information regulator sirtuin 1 (SIRT1)/peroxisome proliferator-activated receptor-γ coactivator 1α (PGC-1α) pathway was activated, whereas the phosphoinositide 3-kinase (PI3K)/protein kinase B (AKT) signaling pathway was inhibited. Additionally, Rb1 could rescue mitochondrial dysfunction by promoting the mitochondrial fusion protein mitofusion 1 (MFN1) and inhibiting dynamin-related protein 1 (DRP1) expression and apoptosis in the lungs. In BEAS-2B cells, Rb1 plays a role similar to that of a SIRT1 agonist (SRT1720), including enhancing mitochondrial membrane potential and decreasing mitochondrial ROS and DRP1 translocation to mitochondria. Our findings suggest that Rb1 maintains mitochondrial integrity by activating SIRT1/PGC-1α and inhibiting PI3K/AKT, thereby ameliorating asthmatic airway inflammation.

To study the toxicity of fentanyl analogs that damage the liver and kidneys in rats, these analogs were evaluated by examining two types of μ-opioid receptor (MOR) signaling responses using HEK293 cells. The results indicated that, in the MOR-mini-Gi recruitment assay, the 50% effective concentration (EC) ranked as iBF < DAMGO ≈ 4F-iBF < 4Cl-iBF, and the maximum response (E) ranked as iBF ≈ DAMGO > 4F-iBF > 4Cl-iBF. In the MOR-β-arrestin 2 recruitment assay, the EC ranking was DAMGO < iBF < 4F-iBF < 4Cl-iBF, and the E ranking was DAMGO > iBF > 4F-iBF. In addition, each of the desphenethylated metabolites, likely the major metabolites of these analogs, showed no MOR signaling responses.

This study investigated the protective effects of metformin against combined high glucose (HG)- and UVA-induced cytotoxicity in fetal rat skin keratinocytes (FRSK cells), a model of diabetic photoaging. HG combined with UVA caused a synergistic loss of cell viability accompanied by marked increases in phosphorylation of AMP-activated protein kinase (p-AMPK), reactive oxygen species (ROS) generation, senescence-associated β-galactosidase (SA-β-Gal) activity, and Sirtuin 1 (SIRT1) expression. HG alone induced moderate cytotoxicity and senescence, whereas UVA alone under normal glucose conditions (NG + UVA) produced negligible ROS and minimal viability loss. Metformin improved cell viability under dual stress conditions in a dose-dependent manner, with maximal protection observed at 8 mM. In UVA-free cultures, metformin increased p-AMPK in both NG and HG, peaking at 8 mM. Under HG + UVA, p-AMPK was higher than in NG + UVA and HG alone, with no additional increase following metformin treatment. ROS accumulation occurred only under HG + UVA and was strongly suppressed by metformin, nearly to baseline at 8 mM. The HG + UVA-induced increases in SA-β-Gal activity and SIRT1 expression were reduced in parallel with ROS suppression. These findings suggest that metformin's cytoprotective effect in this model is primarily mediated by attenuation of ROS rather than by further AMPK activation, indicating an AMPK-independent antioxidant mechanism.

It is known that the daily feeding cycle affects the dosing time-dependent changes in the pharmacodynamics and pharmacokinetics of many drugs. Our previous study demonstrated that administration of empagliflozin (EMPA), sodium-glucose cotransporter 2 (SGLT2) inhibitor, at the beginning of daily feeding cycle (active phase) effectively prevents the development of neuropathic pain in streptozotocin (STZ)-induced diabetic mice. Although the blood glucose levels are closely related to feeding, the relationship between the daily feeding cycle and the optimal dosing time of EMPA remains unclear. In this study, we used STZ-induced diabetic mice and implemented a daily time-restricted feeding (TRF) regimen to investigate whether the dosing time-dependent preventive effect of EMPA on the diabetic neuropathy is modulated by TRF. Animals were housed under a 12-h light/dark cycle, and were assigned to either light-phase TRF (feeding during the light phase) or dark-phase TRF (feeding during the dark phase). The hypoglycemic effect of EMPA was enhanced when the drug was administrated at the beginning of both TRF conditions. A similar influence of the daily feeding cycle on the dosing time-dependent hypoglycemic effect of EMPA was also observed in its preventive effect on the development of diabetic neuropathic pain. Further analysis revealed that dosing time-dependent variations in both the hypoglycemic effect of EMPA and its preventive effect on diabetes-induced pain hypersensitivity were attributable to corresponding changes in urinary glucose excretion. Our results support the notion that the administration of EMPA at the onset of daily feeding cycle effectively suppresses the development of diabetic peripheral neuropathy.

In vitro studies have reported that multiple vitamins may promote bacterial growth and infection. In clinical practice, patients receiving peripheral parenteral nutrition (PPN) may develop catheter-related bloodstream infections (CRBSIs). Since some PPN formulations contain multiple vitamins, they may increase the risk of CRBSIs. Therefore, the present study investigated the relationship between PPN infusions containing vitamins and the incidence of CRBSIs and examined the effects of different antiseptics used before catheter insertion. Patients were divided into the following groups: those receiving the BFLUID injection (1 vitamin), those receiving the PAREPLUS injection (9 vitamins), those disinfected with povidone iodine before the PAREPLUS injection, and those disinfected with chlorhexidine alcohol before the PAREPLUS injection. We analyzed infection rates in each group. The PAREPLUS group showed a significantly higher rate of infections by Staphylococcus species. Furthermore, a longer duration of PPN administration (≥2 weeks) was associated with a higher risk of CRBSI across all detected pathogens. Vitamin-enriched PPN increased approximately 2-fold in CRBSI compared with a thiamine-only formulation, independent of infusion duration. However, the incidence of CRBSI did not significantly differ between the different antiseptic groups. These results suggest that proper infection control and careful management are essential during PPN therapy, especially when multiple vitamin products are used over extended periods.

"Antibody engineering" is a promising strategy for generating high-affinity antibodies required for developing sensitive immunoassays. Therein, the variable domains (V and V) of the parental antibody are genetically randomized and combined to produce diverse single-chain Fv fragment (scFv) molecules. Subsequently, high-affinity scFv mutants are selectively isolated. In the randomization process, mutations have conventionally been targeted to the complementarity-determining regions (CDRs) in the variable domains, which often interact directly with antigens. However, we previously discovered that, pinpoint insertion of only a single amino acid (leucine, asparagine, aspartic acid, proline, glutamine, arginine, or histidine) between positions 6 and 7 in the framework region 1 (FR1) of the V, which is unlikely to interact with antigens, enhanced the affinity of an anti-cortisol scFv (original K, 3.6 × 10 M) up to 17-61-fold. These findings prompted us to conduct a comprehensive study of this affinity-enhancement phenomenon involving the remaining amino acids. Thus, we generated the necessary 13 scFv mutants and compared their K values. Remarkably, all mutants showed enhanced affinities, similar to those of the previous 7 mutants. Among the 20 mutants, the leucine-inserted scFv showed the largest K (2.2 × 10 M) and consequently enabled a 75-fold more sensitive enzyme-linked immunosorbent assay (midpoint, 9.86 pg/assay) compared to the assay using the parental scFv (midpoint, 744 pg/assay). In silico modeling suggested that, regardless of the amino acid inserted, elongated FR1 can alter the conformation of the CDR3 in V to facilitate a favorable interaction with cortisol.

Pregabalin is a first-line treatment for neuropathic pain, but its oral use is often limited by central nervous system side effects, which may require dose reduction or discontinuation. A transdermal formulation may help minimize this adverse effect. The aim of this study was to improve the skin permeability of pregabalin by assessing the feasibility of in-hospital preparation and evaluating its stability in the early stages of formulation development. A 0.4% pregabalin formulation was prepared using hydrophilic cream, lipophilic cream, and pluronic lecithin organogel (PLO) gel. Ex vivo skin permeation studies were conducted using Franz diffusion cells and excised dorsal skin from hairless male mice (HR-1, 7 weeks old). Samples were collected over time and analyzed by LC-tandem MS. Drug content, pH, and viscosity were assessed under 2 storage conditions (25°C/60% relative humidity [RH] and 40°C/75% RH) to evaluate stability. The hydrophilic cream showed the highest cumulative drug permeation, flux, and permeability coefficient. Penetration enhancers added to the PLO gel did not improve absorption. Drug content remained stable under 25°C/60% RH. An inverse correlation was observed between viscosity and drug permeation, suggesting that lower viscosity facilitated diffusion. pH values remained within the physiological range, indicating low irritation potential. The hydrophilic cream demonstrated superior transdermal delivery characteristics for pregabalin. These findings support its potential as an effective and safe alternative to oral administration for managing neuropathic pain.

This study established an orthotopic mouse model of colorectal cancer (CRC) liver metastasis to investigate the inhibitory effects of combined phytic acid (IP6) and inositol (INS) at different ratios on CRC liver metastasis, as well as their impact on the phosphatidylinositol 3-kinase/serine/threonine protein kinase (PI3K/AKT) signaling pathway and macrophage polarization. Combined treatment with varying ratios of IP6 and INS significantly enhanced survival rates and reduced both cecal tumor weight and the incidence of liver metastasis. Flow cytometry indicated an increased CD4/CD8 T cell ratio and a decrease in regulatory T cells, with the 1 : 3 group showing the most pronounced effects (p < 0.05). Cytokine levels were modulated after treatment with varying IP6-to-INS ratios, with a significant reduction in CCL20, interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α) (p < 0.05), and an increase in IL-10 and transforming growth factor-β (TGF-β) (p < 0.05). Western blot analysis confirmed a significant downregulation of PI3K/AKT pathway proteins, including PI3K, phosphorylated (p)-PI3K, AKT, p-AKT, and mammalian target of rapamycin (p < 0.05), with the 1 : 3 group exhibiting the greatest effect. Additionally, the expression of M2 macrophage polarization-related proteins CD163 and CD206 was downregulated (p < 0.05). Our findings suggest that the combination of IP6 and INS alleviates CRC metastasis by downregulating the PI3K/AKT pathway and influencing macrophage polarization, with the most significant inhibitory effect observed at an IP6-to-INS ratio of 1 : 3.