ABSTRACTGlioblastoma multiforme (GBM) is among the most aggressive brain tumors, largely due to the restrictive blood–brain barrier (BBB) and limited drug bioavailability. Temozolomide (TMZ), an alkylating agent, and perillyl alcohol (POH), a monoterpene with antitumor properties, have shown promise in GBM therapy. This study developed and validated a UPLC‐MS/MS method for the simultaneous quantification of TMZ, its active metabolite 5‐aminoimidazole‐4‐carboxamide (AIC), and perillic acid (PA), the primary POH metabolite, in rat plasma and tissues. The method met FDA and EMA validation criteria, showing high selectivity, linearity (R2 > 0.995), accuracy (80%–120%), precision (RSD < 15%), and matrix‐specific stability across plasma, brain, liver, kidney, spleen, and lung. Following single oral doses of TMZ (2 mg/kg) and POH (47 mg/kg), pharmacokinetic analysis revealed that TMZ had the highest systemic exposure (AUC0–24h: 8173.64 ng·h/mL) and Cmax (1448.64 ng/mL), while PA showed the fastest absorption (tmax: 0.5 h). AIC levels confirmed efficient TMZ metabolism. Biodistribution analysis showed TMZ accumulation in the brain (~1000 ng/mL), supporting its CNS efficacy. PA was mainly distributed to liver and kidneys, with limited brain penetration. The validated method enables preclinical pharmacokinetic and tissue distribution studies, offering valuable insights into TMZ and POH behavior for GBM treatment.

Compartmentalized orthogonal biosynthesis for boosting limonene and oxygenated monoterpenoid production in Saccharomyces cerevisiae.

Background: Limonene, a monocyclic monoterpene abundantly found in Eucalyptus oil and citrus fruits, has gained increasing attention for its potential anticancer properties. Limonene may be safe and well-tolerated while suppressing tumor development, inducing apoptosis, and modulating many molecular pathways, according to preclinical and clinical studies.It is a promising candidate for cancer prevention and treatment due to its natural origin, bioavailability, and selective accumulation in lipid-rich tissues.With an emphasis on its modes of action, safety profile, and therapeutic potential across a range of cancer models, the current review attempts to assess the concentration, dosage, and anticancer effects of limonene extract from eucalyptus oil.  Materials and Methods: A thorough search was conducted using databases such as PubMed, Scopus, and Google Scholar to gather pertinent studies on limonene's anticancer effects. In vitro, in vivo, and clinical investigations were analyzed to determine its bioactivity, tissue distribution, effective dosage ranges, and molecular interactions. The collected data was examined and presented to provide a full understanding of the role of limonene in cancer treatment.  Results: Limonene has shown considerable anticancer action in a dose-dependent manner by inducing apoptosis, autophagy, and arresting the G2/M cell cycle. It affects key pathways like caspase activation, Ras signaling, p53 regulation, and oxidative stress mitigation. Clinical investigations have shown its safety, with little gastrointestinal symptoms noted at therapeutic levels. Combination therapy with drugs like curcumin and andrographolide improves efficacy, whereas derivatives like perillyl alcohol improve potency and selectivity.  Conclusion: Limonene is a safe, bioactive, multifunctional chemical with powerful anticancer properties. However, variations in concentration, dose, and formulation indicate the need for more standardized clinical trials to optimize delivery methods and establish their therapeutic significance in cancer treatment.

Abstract The blood-brain barrier (BBB) is a selective, protective barrier that serves to shield the brain from harmful substances while regulating transport of essential molecules. While this barrier is crucial for maintaining the brain’s homeostasis, it presents a significant challenge for drug delivery, particularly for treating central nervous system (CNS) disorders. Many therapeutic agents, especially large or hydrophilic molecules, are unable to cross the BBB effectively. This includes a wide range of antibiotics, anticancer agents, monoclonal antibodies, and most biologics. Even small molecule drugs can be actively pumped out by efflux transporters like P-glycoprotein, reducing their CNS bioavailability. Consequently, numerous potentially effective drugs fail to reach therapeutic concentrations in the brain. This limitation has major implications for treating brain malignancies, such as glioma. For example, the chemotherapy drug doxorubicin, while effective against many cancers, is largely excluded from the brain due to the BBB. To overcome this obstacle, we have investigated the potential of intra-nasal co-administration of therapeutic drugs with NEO100 (a highly pure, pharmaceutical-grade form of the natural monoterpene perillyl alcohol) to enhance brain delivery through direct nose-to-brain transport. We have shown that NEO100 significantly enhances the brain tumor specific uptake and retention of doxorubicin, dexamethasone, and remdesivir when administered via an intra-nasal route in a formulation that contains NEO100 at concentrations as low as 1%, when compared to traditional oral or IV routes of administration. The evaluation of both drugs alone and in the presence of NEO100 was performed by quantitative HPLC analysis at time points ranging from 15 minutes to 24 hours post administration. Understanding how NEO100 functions in manipulating the barrier’s physiology is crucial for developing next-generation therapeutics capable of treating CNS diseases effectively. Continued innovation in drug design and delivery technologies is essential for expanding the range of treatable neurological conditions

Abstract Glioblastoma multiforme (GBM) remains one of the most lethal and treatment-resistant brain tumors due to its diffuse infiltration, poor response to standard therapies, and protection by the intact blood-brain barrier (BBB). Sonodynamic therapy (SDT), which combines low-intensity focused ultrasound (FUS) with ultrasound-responsive drugs (sonosensitizers), offers a promising, noninvasive strategy to overcome these limitations. However, the discovery and validation of effective sonosensitizers suitable for clinical translation has been constrained by traditional in vitro models that are low-throughput and poorly mimic tumor microarchitecture. To address this gap, we developed an integrated screening platform combining artificial intelligence (AI)-guided drug prioritization with high-throughput 3D bioprinted GBM spheroid models. This approach enables systematic evaluation of BBB-penetrant candidates under clinically relevant ultrasound parameters. We focused on NEO100, a Phase 2a investigational perillyl alcohol derivative with established BBB permeability, and NEO212, a Phase 1 novel conjugate of NEO100 and temozolomide (TMZ). Both were assessed for their potential as sonosensitizers using magnetically printed 3D GBM cultures and sonication protocols aligned with translational FUS applications. This platform represents a scalable and physiologically relevant method to identify ultrasound-responsive compounds for SDT. Given NEO100’s clinical status and favorable pharmacological properties, this methodology may accelerate the path toward personalized SDT strategies in GBM and other brain malignancies for patients.

Exploring the hepatoprotective effects of perillyl alcohol in the experimental models of acute liver injury.

Perillyl alcohol ameliorates high-fat diet-induced obesity in mice by modulating gut microbiota and activating IRF4-mediated brown fat thermogenesis.

IntroductionCymbopogon martinii (Roxb.) Wats. essential oil (CMEO) exhibits significant variation in composition and bioactivity across different growth stages. Understanding these changes is crucial for optimizing its therapeutic and industrial applications.MethodsCMEO was extracted at the vegetative, reproductive, and post-reproductive stages using hydro-distillation. Chemical composition was analyzed by GC-MS. Antimicrobial activity was assessed using disc diffusion and direct contact assays, while antioxidant potential was evaluated through DPPH, ABTS, and β-carotene bleaching assays. Correlation analysis was performed to link major bioactive compounds with biological activities.ResultsA total of 59 compounds were identified, with the reproductive stage showing the highest diversity (49 compounds, 97.86%) and oil yield. Major compounds varied across stages: carveol (20.87%), trans-p-mentha-1(7),8-dien-2-ol (12.9%), and D-limonene (6.2%) dominated the vegetative phase; cis-piperitol (15.27%), cis-p-mentha-1(7),8-dien-2-ol (15.52%), and carvone (3.31%) were abundant in the reproductive phase; while the post-reproductive phase was rich in trans-p-mentha-1(7),8-dien-2-ol (19.58%) and carveol (11.32%). Antibacterial and antifungal activities were highest during the reproductive stage, particularly against Staphylococcus aureus and Alternaria alstroemeriae. Antioxidant potential peaked during the post-reproductive phase, with the lowest IC50 values.DiscussionCorrelation analysis revealed negative associations between key bioactive compounds (e.g., carvone, D-limonene, α-methylcinnamaldehyde, and (S)-perillyl alcohol) and microbial/oxidative inhibition thresholds, confirming their contribution to CMEO bioactivity. These findings highlight the critical role of harvest timing in maximizing the chemical richness, antimicrobial efficacy, and antioxidant potential of CMEO.

ABSTRACTIn this study, (S)‐4‐((4‐(prop‐1‐en‐2‐yl)cyclohex‐1‐enyl)methoxy)phthalonitrile, as a novel phthalonitrile derivative, which is the starting material for phthalocyanine synthesis, was prepared and characterized. Then, its symmetric metal‐free [H2Pc (2)] and metallophthalocyanine derivatives [ZnPc (3), CuPc (4), and CoPc (5)] were successfully synthesized, and their structures were characterized with the combination of suitable spectroscopic methods. The solubility and aggregation properties of novel phthalocyanines were investigated. Also, the photophysical and photochemical properties of the newly synthesized H2Pc (2) and ZnPc (3) were studied in solution media. The singlet oxygen quantum values are 0.45 for H2Pc (2) and 0.75 for ZnPc (3). According to these results, the peripheral perillyl alcohol substitution of phthalocyanines increased the therapeutic efficacy. At the same time, the newly synthesized phthalocyanines (2–5) displayed the strongest inhibitory effect. The results indicate a high affinity and selectivity for carbonic anhydrases. This underscores their potential as promising lead compounds for developing therapeutic agents to treat conditions such as glaucoma, epilepsy, and certain types of cancer.

Evaluation of the efficacy of Perillyl alcohol in the treatment of Eimeria tenella infection.

Perillyl alcohol (POH) is a natural compound presented in essential oils extracted from plants, recognized for its capacity to stimulate apoptosis in tumor cells while leaving normal cells unharmed. The present study has established a square‐wave adsorptive anodic stripping voltammetric (SW‐AdASV) technique for the detection of POH in both bulk, and biological fluids, utilizing an activated graphite sensor (AGPS). The electrochemical oxidation performance of POH was studied via cyclic voltammetry and subsequently optimized with SW‐AdASV technique. Under the standard operational conditions, linear calibration curves for POH were obtained within concentration ranges of 0.001–0.013 nM and 0.003–0.018 nM. The detection limits (LODs) for bulk and human serum were evaluated to be 0.3 pM and 0.9 pM, respectively. Moreover, The AGPS effectively detected POH concentrations in human serum during routine analyses, with no interference from other serum components. Additionally, it demonstrated satisfactory repeatability, reproducibility, and a storage stability of 94.54%.

Glioblastoma is the most aggressive and fatal primary brain tumor in adults, characterized by poor prognosis and limited treatment efficacy due to the impermeability of the blood-brain barrier (BBB) and its treatment-resistant nature. This review aims to evaluate the potential of intranasal terpene treatment (ITT) as a novel and non-invasive strategy to bypass the BBB and improve glioblastoma treatment outcomes. A review of recent preclinical and clinical studies on intranasally administered compounds (especially terpenes such as Perillyl alcohol (POH)) is presented in terms of their molecular mechanisms, bioavailability, and clinical effects in the central nervous system (CNS).

Osteoarthritis (OA) is a common degenerative disease characterized by chondrocyte death and extracellular matrix (ECM) degradation, but the present OA treatments are only symptomatic and not aetiologic. Autophagy is a key protective mechanism for OA through the inhibition of chondrocyte apoptosis and degeneration. Perillyl alcohol (POH) is a monomer derived from Chinese herbal medicines that can regulate autophagy and has anti-inflammatory effects. In this study, cell viability, chondrocyte phenotype and the expression of molecules involved in autophagy and the PI3K-AKT-mTOR signalling pathway were detected to explore the hypothesis that POH may decrease the progression of OA by regulating autophagy. We found that POH significantly inhibited the expression of inflammatory cytokines, maintained the chondrocyte phenotype, and protected against ECM degeneration. We also provide evidence that the anti-inflammatory effect of POH works by regulating autophagy via the PI3K-AKT-mTOR signalling pathway. This study demonstrates for the first time that POH has a therapeutic effect on arthritis, which suggests that POH may be a promising candidate for OA therapy.

This work focused on developing CS/PPy, a pH-responsivecontrolled-releasenanocarrier composed of natural polymer, the chitosan (CS), and syntheticpolymer, the polypyrrole (PPy). The goal was to evaluate its potentialapplication in controlled release studies of perillyl alcohol (POH).To enhance its properties, CS/PPy was cross-linked with glutaraldehyde,resulting in a new material, CS/PPy/GA. CS/PPy and CS/PPy/GA demonstratedhigh encapsulation efficiency for POH. The release of POH was investigatedat pH 4.5, 6.0, and 7.4. The POH release rate exhibited pH-dependentbehavior, with a significantly higher release observed in acidic media(pH 4.5) compared to neutral/basic media (pH 7.4). This suggests thepotential for controlled release of POH in response to specific physiologicalenvironments. The Korsmeyer-Peppas model was followed for the releaseof POH from CS/PPy at all pH values that were studied. The releasebehavior of POH from CS/PPy/GA exhibited a pH-dependent mechanism.The Korsmeyer-Peppas model best described release at pH 4.5 and 6.0,while the Higuchi model provided a better fit at pH 7.4. The resultssuggested that the synthesized materials exhibited high affinity forPOH, and the release behavior is pH-dependent.

Recent years have seen increased focused on developing new anti-parasitic drugs that are both highly effective and low in toxicity, as the widespread use of existing anti-parasitic drugs has raised growing concerns. Natural products have gained significant interest due to their diverse biological activities with minimal toxic side effects. Our previous studies have already demonstrated the good anti-E. tenella effect of Perillyl Alcohol. To further investigate its efficacy against other protozoa, we selected Toxoplasma gondii as the researchsubject. In this study, we utilized the CCK-8 assay in vitro to assess the cytotoxicity of Perillyl Alcohol on DF-1 cells. The impact of Perillyl Alcohol of T. gondii tachyzoite invasion and intracellular proliferation were investigated in vitro. In vivo, we evaluated the effect of Perillyl Alcohol on the pathogenicity of T. gondii, including host survival time, liver and spleen tissue damage, cysts formed in the brain. Furthermore, we analyzed the expression levels of isoprenylation-related genes using quantitative PCR (qPCR). The half maximal inhibitory concentration (CC50) of Perillyl Alcohol against DF-1 cells was determined to be 525.0. In vitro studies showed that treatment with Perillyl Alcohol effectively inhibited the invasion rate and intracellular proliferation of T. gondii tachyzoite. The half maximal inhibitory concentration (IC50) was calculated to be 314.3, and further analysis yielded a selectivity index (SI) of 1.67. In vivo, Perillyl Alcohol treatment prolonged the survival time and increased the survival rate of T. gondii-infected mice, while reducing the parasite burden in liver and spleen tissues. It also demonstrated a certain protective effect against T. gondii-induced tissue damage, including effectively alleviating hepatosplenomegaly and mitigating the elevation of serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels induced by hepatic injury. Building on this foundation, we further explored the impact of Perillyl Alcohol on the formation of brain cysts and found that it could significantly reduce the number of brain cysts induced by the Pru strain of T. gondii infection. After treatment with Perillyl Alcohol, the expression levels of isoprenylation-related enzymes Tgmecs, Tgdxr, and Tghdr were significantly reduced. These results demonstrate that Perillyl Alcohol may suppress the growth of T. gondii by significantly inhibiting the expression of enzymes involved in isoprenoid biosynthesis.

Neuroprotective effect of Perillyl alcohol in sporadic Alzheimer's disease in rats.

ABSTRACTHaematological cancer types, such as leukaemia and lymphoma, represent diseases that are life‐threatening to canine and human patients alike, and better treatments are needed. We are developing a novel anticancer agent, NEO212, a conjugate of two cancer drugs, the alkylating agent temozolomide (TMZ) and the monoterpene perillyl alcohol (POH). NEO212 has revealed robust therapeutic activity in preclinical tumour models harbouring different human cancer types. In the comparative preclinical study presented here, a two‐species (canine and human) and two‐cancer (leukaemia and lymphoma) analysis was performed to determine whether the promising therapeutic activity of NEO212 would span species and cancer types. We investigated the activity of NEO212 in human and canine leukaemia and lymphoma cell lines in vitro and in corresponding mouse models in vivo. Our results show that in vitro NEO212 is significantly more potent than TMZ and POH in all cell lines and exerts activity even against strongly TMZ‐resistant tumour cells. In vivo, oral NEO212 strikingly extends the survival of mice harbouring human or canine leukaemia or lymphoma cells. At the same time, NEO212 is well tolerated in dogs at dosages higher than those that achieved therapeutic activity in mouse models. Our study introduces NEO212 as a novel oral cancer drug candidate for both human and veterinary oncology applications.

As a result of the study on antimicrobial activity it was established that the essential oil of the herb of the drooping dragonhead (Dracocephalum nutans L.) exhibits pronounced antimicrobial activity in relation to strains of gram-positive bacteria Staphylococcus aureus (ATCC 6538), Bacillus subtilis (ATCC 6633) and weak activity in relation to gram-negative bacteria Escherichia coli (ATCC 25922), Pseudomonas aeruginosa (ATCC 27853) and weak activity to the yeast fungus Candida albicans (ATCC 10231). Taking into account the above, the purpose of our research is to develop a gel with antimicrobial properties. The essential oil of Dracocephalum nutans L. was chosen as the object of the study. The genus Dracocephalum L. belongs to the family Lamiaceae Lindl. (Labiatae), the plants of which are of interest as sources of medicinal preparations. Studies have shown that some species of Dracocephalum possess antibacterial, antitussive, antidiarrheal, antioxidant, anticancer, anti-inflammatory, antidiabetic and sedative properties. Isolated compounds from the genus Dracocephalum collected from natural ones such as limonene, carvacrol, α-pinene, γ-terpinene, perillaldehyde, caryophyllene oxide, 1,8-cineole, verbenone, perillyl alcohol, neral and geranyl acetate have pronounced antimicrobial properties. Among the plants of this genus, Dracocephalum nutans L. is of interest. Dracocephalum nutans L. (Synonyms: Dracocephalum nutans var. alpinum Kar. &amp; Kir., Dracocephalum nutans subsp. subarctium Kuvaev) contains a complex of biologically active substances and is used in the medicine of the peoples of Southeast Asia in the treatment of kidney inflammation and gastrointestinal diseases such as hepatitis, gastritis, etc. The herb of Dracocephalum nutans L. has a high content of various biologically active compounds that determine the medicinal properties of this species: essential oils, flavonoids, saponins, coumarins, traces of alkaloids. The main natural components of the essential oil of Dracocephalum nutans L. were 1,8-cineole (34%), α-pinene (6.7%), β-pinene (5.2%), β-myrcene (5.3%). The chemical composition of the essential oil was studied by chromatography-mass spectrometry. A unified method for determining the component composition of essential oils, as well as a 7890A Agilent Technologies chromatographic system with a 5975C mass spectrometric detector were used for the analysis. Essential oils have various biological activities: antimicrobial, anti-inflammatory, immunomodulatory, anesthetic. There is data on the use of essential oils as an activator of absorption of pharmacologically active components of the dosage form. One of the main properties of essential oils is a high antimicrobial effect, combined with low toxicity and the absence of side effects in therapeutic use. It is assumed that the mechanism of the antimicrobial action of essential oils lies in the destruction of membrane structures of microbial cells with subsequent disruption of intracellular metabolism and death of microorganisms.

PROTOCOL OF A RANDOMIZED PILOT STUDY ON SURVIVAL IN NEWLY DIAGNOSED GLIOBLASTOMA PATIENTS UNDERGOING CHEMORADIATION VERSUS COMBINED CHEMORADIATION WITH INTRANASAL PERILLYL ALCOHOL

Avocado is a rich source of numerous nutrients, such as micro- and macroelements, essential unsaturated fatty acids, and vitamins essential for the correct functioning of the body. Consequently, its consumption has significantly increased in recent years. The primary edible part of the fruit is the flesh, while the seed is still considered biowaste. Currently, various methods for utilization of this biowaste are being explored, prompting the authors of this work to investigate the catalytic properties of ground avocado seeds. Dried, ground avocado seeds were used as the catalyst in the environmentally friendly oxidation of limonene with oxygen. The process was carried out in mild conditions, without the use of any solvent and at atmospheric pressure. The studies examined the influence of temperature (70–110 °C), the amount of the catalyst (0.5–5.0 wt%), and the reaction time (15–360 min). The analyses of the post-reaction mixtures were performed using the gas chromatography method (GC). The maximum value of the conversion of limonene obtained during the tests was 36 mol%. The main products of this process were as follows: 1,2-epoxylimonene, carveol, and perillyl alcohol. Also, the following compounds were determined in the post-reaction mixtures: carvone and 1,2-epoxylimonene diol. The studied process is interesting, taking into account both the management of waste in the form of avocado seeds and possible wide applications of limonene transformation products in medicine, cosmetics and the food industry. Given that limonene is now increasingly being extracted from waste orange peels, this is also a good way to manage the future naturally derived limonene and reduce the amount of waste orange peels. The presented studies fit perfectly with the goals of sustainable development and circular economy and may be the basis for the future development of “green technology” for obtaining value-added oxygenated derivatives of limonene. These studies show the use of waste biomass in the form of avocado seeds to obtain a green catalyst. In this context, our research presents an effective way of waste valorization.