Good Penetration Research Articles

Evaluation of Permeability, Safety, and Stability of Nanosized Ketoprofen Co-Spray-Dried with Mannitol for Carrier-Free Pulmonary Systems

Pulmonary drug delivery presents a promising approach for managing respiratory diseases, enabling localized drug deposition and minimizing systemic side effects. Building upon previous research, this study investigates the cytotoxicity, permeability, and stability of a novel carrier-free dry powder inhaler (DPI) formulation comprising nanosized ketoprofen (KTP) and mannitol (MNT). The formulation was prepared using wet media milling to produce KTP-nanosuspensions, followed by spray drying to achieve combined powders suitable for inhalation. Cell viability and permeability were conducted in both alveolar (A549) and bronchial (CFBE) models. Stability was assessed after storage in hydroxypropyl methylcellulose (HPMC) capsules under stress conditions (40 °C, 75% RH), as per ICH guidelines. KTP showed good penetration through both models, with lower permeability through the CFBE barrier. The MNT-containing sample (F1) increased permeability by 1.4-fold in A549. All formulations had no effect on cell barrier integrity or viability after the impedance test, confirming their safety. During stability assessment, the particle size remained consistent, and the partially amorphous state of KTP was retained over time. However, moisture absorption induced surface roughening and partial agglomeration, leading to reduced fine particle fraction (FPF) and emitted fraction (EF). Despite these changes, the mass median aerodynamic diameter (MMAD) remained stable, confirming the formulation’s continued applicability for pulmonary delivery. Future research should focus on optimizing excipient content, alternative capsule materials, and storage conditions to mitigate moisture-related issues. Hence, the findings demonstrate that the developed ketoprofen–mannitol DPI retains its quality and performance characteristics over an extended period, making it a viable option for pulmonary drug delivery.

208. Exposure-response relationship of ceftazidime/avibactam in adults with central nervous system infections caused by carbapenem-resistant Klebsiella pneumoniae: a prospective observational study

Abstract Background Central nervous system (CNS) infections caused by carbapenem-resistant Klebsiella pneumoniae (CRKP) poses significant challenges for clinicians in ICU. Achieving optimal drug exposure in the cerebrospinal fluid (CSF) is crucial for good outcomes. This study aimed to evaluate the exposure-response relationship of ceftazidime/avibactam (CZA) in plasma and CSF. Methods All patients received CZA (2.5 g), IV infusion (over 2 hours), q8h. At steady-state after the fourth dose of CZA, the concentrations of CZA in the plasma and CSF were determined by ultra-performance liquid chromatography (UPLC) at 1, 2, 2.25, 2.5, 3, 4, 6, and 8 h post-dose. The PK/PD index of ceftazidime (CAZ) and avibactam(AVI) was calculated for each patient. Demographic, clinical efficacy, microbiological eradication, and 28-day mortality were reviewed. Blood and CSF samples were collected for calculation of PK parameters by non-compartmental pharmacokinetics (PK) modeling. The attainment of CAZ and AVI PK/PD targets in plasma and CSF was calculated to assess the exposure-response relationship of CZA and the corresponding clinical efficacy rate and bacterial eradication rate. Results CAZ and AVI demonstrated good penetration into CSF, evidenced by median CSF/plasma AUCtau ratio of 43.8 % and 55.8 %, respectively. CZA 2.5 g q8h achieved CAZ PK/PD target of 50%fT >MIC in both plasma and 50%T >MIC in CSF of all patients and reached AVI PK/PD target of 50%T >CT=1 mg/L in CSF of 80% (16/20) of patients. The dosing regimen also achieved CAZ PK/PD target of 100% fT >4MIC in plasma of 9 patients and 100% T >4MIC in CSF of 14 patients. At the end of the treatment, the overall clinical and microbiological efficacy rates were 80.0% and 100%, respectively. Only one case of CZA-related adverse event (Clostridioides difficile-associated diarrhoea) was reported. Conclusion The good CSF penetration of CZA enables the dosing regimen of CZA 2.5 g q8h a promising treatment option for CNS infections caused by CRKP. Disclosures Nanyang Li, Master of Medicine, TenNor Therapeutics (Suzhou) Ltd: Investigator Jing Zhang, Doctor of Medicine, TenNor Therapeutics (Suzhou) Ltd: Investigator

Radiation-induced aerobic oxidation via solvent-derived peroxyl radicals.

Oxidation is a fundamental transformation in synthesis. Developing facile and effective aerobic oxidation processes under ambient conditions is always in high demand. Benefiting from its high energy and good penetrability, ionizing radiation can readily produce various reactive species to trigger chemical reactions, offering another option for synthesis. Here, we report an ionizing radiation-induced aerobic oxidation strategy to synthesize oxygen-containing compounds. We discovered that molecular oxygen (O2) could be activated by reactive particles generated from solvent radiolysis to produce solvent-derived peroxyl radicals (RsolOO·), which facilitated the selective oxidation of sulfides and phosphorus(iii) compounds at room temperature without catalysts. Density functional theory (DFT) calculations further revealed that multiple RsolOO· enable the oxidation reaction through an oxygen atom transfer process. This aerobic oxidation strategy broadens the research scope of radiation-induced chemical transformations while offering an opportunity to convert nuclear energy into chemical energy.

Development of a curcumin-piperine nanoparticle system using dissolving microneedles for transdermal drug delivery in malaria treatment: In vitro evaluation.

The combination of the active compounds curcumin and piperine (CP) is effective as an antimalarial; however, the solubility and bioavailability of CP are very low. This study aims to formulate CP in nanoparticles (NP), which are then fabricated into dissolving microneedles (DMN). The NPs were prepared with a concentration ratio of CP-Chitosan-So.TPP-So.Alginate (0.1:0.04:0.02:0.03). Subsequently, NPs-CP-DMN were formulated with NPs-CP concentrations (35:40:50 w/w) and a mixture of the polymers polyvinyl alcohol (PVA) and polyvinylpyrrolidone (PVP) in a ratio of (35:65, 40:60, 50:50). Characterization of the nanoparticles and microneedles was conducted, including dissolution time tests, permeation studies, hemolysis assessment, dermatokinetics, and in vitro antiplasmodial activity testing. The results showed that NPs-CP had an average size of 446.67 ± 40.27 nm and 367.6 ± 26.31 nm. On the formula NPs-CP-DMN the addition of PVA and PVP polymers (F2) resulted in DMNs with good mechanical strength and penetration ability, capable of penetrating five layers of Parafilm®. This formulation completely dissolved in 10 min without leaving any residue, with a curcumin flux value of 25.7 ± 0,51 µg/mL and piperine flux of 28.5 ± 0,51 µg/mL. The formulation showed no toxicity, with a hemolysis percentage of < 5 %, Tmax of 7 h, and Cmax values of 11.07 ± 0.31 µg/cm3 for curcumin and 17.40 ± 3.3 µg/cm3 for piperine. Moreover, this formulation effectively inhibited the P.falciparum FCR3 strain parasite, with an IC50 value of 35.9 μg/mL. Therefore, this study holds promise as a new strategy for malaria treatment.

Selective Tissue Penetration of the Corneal Layers of Cyclosporin 2% Associated with Miglyol in Rabbits.

Purpose: Cyclosporin A (CsA) is a drug used to prevent immune rejection in corneal transplantation. Most grafts performed today are endothelial grafts which are complicated with poor penetration of CsA into the endothelium due to its hydrophobicity. To improve CsA penetration into the corneal a new ocular formulation of CsA 2% with Miglyol was developed and is commercially available. The purpose of this pharmacokinetic study was to determine the concentrations of CsA in all layers of the cornea, in particular in the endothelium after topical administration of CsA 2%-Miglyol in rabbits. Methods: Sixteen rabbits were divided in 4 groups according to the time from the last instillation of CsA 2%-Miglyol to corneal sampling. Rabbit eyes received one drop of CsA twice a day for 5 days. Corneal tissue and plasma samples were collected at 2, 6, 12, and 24 h. CsA concentrations were measured using liquid chromatography-tandem mass spectrometry method. Results: Maximum concentrations (Cmax) of CsA were obtained in corneal tissues within 2 h after the last instillation of CsA 2%-Miglyol, except in the endothelium (12 h). Cmax were 59.75 ± 12.09 ng/mg, 15.66 ± 4.31 ng/mg, 5.17 ± 0.88 ng/mg, and 2.65 ± 0.47 ng/mg for the epithelium, endothelium, anterior stroma and posterior stroma. CsA concentrations remained high over a 24-h in all corneal layers. Conclusions: The topical application of CsA 2%-Miglyol allowed a high concentration of CsA in the corneal endothelium. The good penetration of CsA into the endothelium suggests that this formulation can be effective to prevent endothelial graft rejection.

Light-Directed Self-Powered Metal-Organic Framework Based Nanorobots for Deep Tumor Penetration.

Effective intratumoral distribution of anticancer agents with good tumor penetration is of great practical importance for oncotherapy. How to break the limitation of traditional passive drug delivery relying on blood circulatory system into solid tumors remains a challenge. Herein, a light-directed self-powered nanorobot based on zirconium-based porphyrin metal-organic framework (MOF) is reported for smart delivery of chemodrug and photosensitizer for deep tumor penetration. The MOF-based carrier transporting the therapeutics is able to responsively generate hydrogen sulfide as propulsion gas and uniquely exhibits negative phototaxis in acidic tumor microenvironment. Under red light irradiation, the nanorobots are accordingly propelled from the tumor surface toward deep tumor far away from the exterior light source in a specific deep-into-tumor direction, greatly enhancing tumor penetration. Along with tumor-responsive release and activation of the delivered therapeutics inside tumor tissues, photo-chemotherapy with significantly improved efficacy is achieved.

Discovery of NP3-253, a Potent Brain Penetrant Inhibitor of the NLRP3 Inflammasome.

Activation of the NLRP3 inflammasome in response to danger signals is a key innate immune mechanism and results in the production of the pro-inflammatory cytokines interleukin-1β (IL-1β) and interleukin-18 (IL-18) as well as pyroptotic cell death. Aberrant NLRP3 activation has been linked to many acute and chronic conditions ranging from atherosclerosis to Alzheimer's disease and cancer, and based on the clinical success of IL-1-targeting therapies, NLRP3 has emerged as an attractive therapeutic target. Herein we describe our discovery, characterization, and structure-based optimization of a pyridazine-based series of NLRP3 inhibitors initiating from an high-throughput screening campaign. The scaffold, exemplified by lead molecule NP3-253, has excellent potency and physicochemical and pharmacokinetic properties, including good brain penetration. The establishment of pharmacokinetic/pharmacodynamic relationships in the periphery and central nervous system in mechanistic models facilitates the use of NP3-253 as a tool to further interrogate the biology of NLRP3 in peripheral and neuroinflammatory models.

Development of a PET Probe Targeting Bromodomain and Extra-Terminal Proteins for In Vitro and In Vivo Visualization.

Background: Bromodomain and extra-terminal (BET) proteins are critical regulators of gene transcription, as they recognize acetylated lysine residues. The BD1 bromodomain of BRD4, a member of the BET family, has emerged as a promising therapeutic target for various diseases. This study aimed to develop and evaluate a novel C-11 labeled PET radiotracer, [11C]YL10, for imaging the BD1 bromodomain of BRD4 in vivo. Methods: [11C]YL10 was synthesized and evaluated for its ability to bind to the BD1 bromodomain selectively. PET imaging studies were conducted in mice to assess brain penetration, pharmacokinetics, and selectivity. In vitro autoradiography and blocking experiments were performed to confirm the tracer's specificity for the BD1 domain. Results: [11C]YL10 demonstrated good brain penetration, high selectivity for the BD1 bromodomain, and favorable pharmacokinetics in initial PET imaging studies. In vitro autoradiography and blocking experiments confirmed the specific binding of [11C]YL10 to the BD1 domain of BRD4, further validating its potential as a targeted radiotracer. Conclusions: The development of [11C]YL10 provides a new tool for studying BRD4 bromodomains using PET imaging technology. This radiotracer offers potential advancement in the diagnosis and research of neurodegenerative diseases and related disorders involving BRD4 dysregulation.

Chemical composition, antimicrobial, and antioxidant properties of essential oils from Artemisia herba-alba asso. and Artemisia huguetii caball. from Morocco: in vitro and in silico evaluation

IntroductionMorocco is home to a remarkable diversity of flora, including several species from the Artemisia genus. This study aims to thoroughly examine the chemical composition of essential oils derived from Artemisia species and assess their antibacterial and antioxidant properties through in vitro experiments and in silico simulations.MethodsSamples of Artemisia herba-alba Asso. were collected from Boulemane and Ifrane in Morocco, while Artemisia huguetii Caball. was sampled from Tata, representing regions of the Central Middle Atlas and Western Anti-Atlas. Essential oils were extracted using hydrodistillation, and their chemical composition was analyzed by gas chromatography-mass spectrometry (GC-MS). Antibacterial and antifungal activities were evaluated, and antioxidant properties were assessed using the DPPH assay. In silico predictions of antibacterial and antioxidant activities were performed using computational models.ResultsThe extraction yields varied depending on the geographical origin, ranging from 1.54% to 2.78%. GC-MS analysis revealed significant differences in the chemical composition of the oils from different Artemisia species and regions, with a notable prevalence of oxygenated monoterpenes. Specifically, the oil from Boulemane was rich in thujone, the oil from Ifrane was predominantly composed of camphor, and the oil from Tata contained both camphor and thujone. The oils exhibited stronger antifungal than antibacterial properties, with Enterobacter cloacae strains showing high sensitivity, with minimum inhibitory concentrations (MIC) of approximately 12.5 mg/mL. The Boulemane oil of A. herba-alba displayed the highest antioxidant activity, effectively inhibiting DPPH at a concentration of 13.501 μg/mL.DiscussionThe in silico simulations predicted that the primary compounds in these essential oils, such as davanone, eucalyptol, camphor, and thujone, would exhibit potent antibacterial and antioxidant properties. These compounds were found to have favorable ADMET characteristics, including good blood-brain barrier permeability, gastrointestinal absorption, and skin penetration. Molecular docking studies revealed strong interactions between these compounds and key target proteins, such as NADPH-dependent catalase and dihydrofolate reductase. The stability of the protein-ligand complexes was confirmed by molecular dynamics, with davanone showing a significant impact. Overall, this study provides a comprehensive understanding of the biological potential of Artemisia essential oils, highlighting davanone as a promising molecule for medicinal or pharmaceutical applications.

Experimental and numerical study on jet penetrating multi-layer metal rubber air-spaced composite target

Abstract A multi-layer metal rubber air-spaced composite target was designed. Based on the 140 mm caliber shaped charge, the penetration test and simulation study were carried out. By analyzing the damage of the target plate and the interaction process between the jet and the target plate in the simulation, the interference mechanism of the composite target to the jet is revealed. The equivalent armor thickness and anti-jet penetration ability of the composite target under oblique penetration with large inclination angle are obtained by using the equivalent criterion of residual penetration depth. It is concluded that the multi-layer metal rubber air-spaced composite target can not only reduce the load of the armored vehicle, but also has good anti-jet penetration performance, which can provide support and basis for the design of other rubber composite target structures.

Photodegradation of steroid hormone micropollutants with palladium-porphyrin coated porous PTFE of varied morphological and optical properties.

In flow-through reactors, the photodegradation rate can be improved by enhancing contact and increasing the photocatalyst loading. Both can be attained with a higher surface-to-volume ratio. While previous studies focused on thin membranes (30 - 130 µm) with small pore sizes of 20 - 650 nm, this work employed poly(tetrafluoroethylene) (PTFE) supports, of which pore sizes are in the order of 10 µm, while the porosities and thicknesses are variable (22.5 - 45.3 % and 0.2 - 3 mm, respectively). These porous materials were anticipated to allow a higher loading of porphyrin photosensitisers and better light penetration for subsequent photodegradation of steroid hormone micropollutants via singlet oxygen (1O2) generation. The reactor surface refers to the surface within the PTFE pores, while the reactor volume is the total void space inside these pores. The surface-to-volume ratios between 105 and 106 m2/m3 are higher than those of typical microreactors (103 to 104 m2/m3). The weighted average light transmittance varied from 38 % with the thinnest and most porous support to 4.8 % with the thickest support. Good light penetration combined with minimal absorption by PTFE enhanced the light utilisation of the porphyrins when coated in the porous supports. Changes in the support porosity of the coated supports minimally affected steroid hormone removal, because the collision frequency in the very large pores remained relatively constant. However, varying the support thickness, porphyrin loading (0.3 - 7.7 μmol/g), and water flux (150 - 3000 L/m2.h), hence the resulting hydraulic residence time, influenced the collision frequency and steroid hormone removal. Results showed that the supports did not outperform membranes most likely because the larger pore size in the former limited contact between the hormones and 1O2. From photostability testing of the pristine supports, perfluoroalkyl substances (PFAS) released from the supports were found at 10 - 300 ng/L concentrations during accelerated ageing. While PFAS formation was detectable, the quantities during water treatment operations would be extremely low. In summary, this study elucidates the capability and limitations of porous supports coated with photosensitisers to remove waterborne micropollutants.

Research on the Penetration Performance of Zirconium Liner

Abstract As the core part of the shaped charge warhead, the selection of the material of the liner has an important influence on the formation and penetration ability of the shaped charge jet. In this paper, metal zirconium is used as the material of the liner. The Autodyn-2D numerical simulation platform is used to simulate the jet formation and penetration ability of the zirconium liner under the shaped charge structure. By comparing the penetration depth and opening diameter of the same target with the conventional liner material, the penetration ability of different material liners is analysed. The results show that the zirconium liner has good penetration performance compared with the titanium liner, and has good opening performance compared with the copper liner, and the zirconium liner jet will respectively produce secondary hole expansion phenomenon and secondary necking phenomenon when penetrating thick steel target and thick concrete target; When the zirconium liner penetrates the steel target with high strength. the opening ability will gain, but the penetration ability will be impaired. Moreover, the tiny change of the target thickness has little effect on the residual velocity and the opening diameter of the zirconium liner head; The velocity loss of the jet head of the zirconium liner is closely related to the strength of the target plate and the thickness of the target plate. This relationship makes the zirconium liner more conducive to the penetration of medium strength and medium thickness targets.

Revealing inducible clindamycin resistance in methicillin-resistant S aureus : A vital diagnostic imperative for effective treatment.

The World Health Organization added methicillin-resistant S aureus (MRSA) to the list of "priority pathogens," given its capacity to cause life-threatening infections. Clindamycin is a preferred treatment for non-complicated S aureus-induced skin and soft tissue infections. Its good tissue penetration and oral absorption make it suitable for outpatient therapy. However, the emergence of inducible and constitutive (MLS B ) resistance led to clinical challenges, primarily due to the potential oversight of inducible resistance in routine antimicrobial sensitivity testing. This cross-sectional study was conducted at a tertiary care hospital during 2020-2022. A total of 158 MRSA isolates from various clinical specimens were analyzed. The Kirby-Bauer disk diffusion method using cefoxitin disk and D-test were used to identify MRSA and detect inducible clindamycin resistance (ICR), respectively. Among the 158 MRSA isolates, 34.17% showed constitutive clindamycin resistance (MLS B c), while 22.15% displayed ICR (MLS B i). In addition, 10.13% of isolates demonstrated the MS phenotype, clindamycin, and erythromycin susceptibility, with 53 (33.54%) isolates susceptible to both antibiotics. The relative risk of clindamycin treatment failure was 7.66 times higher if the D-test was not used. To prevent clindamycin treatment failures, the D-test must be implemented to detect ICR in MRSA isolate. Neglecting simple and cost-effective tests may lead to inaccurate susceptibility reporting, jeopardizing treatment success.

Discovery of Thiochroman Derivatives as Potent, Oral Selective Estrogen Receptor Degraders and Antagonists for the Treatment of Endocrine-Resistant Breast Cancer.

Selective estrogen receptor degraders (SERDs) deplete the ER signaling pathway via antagonism and degradation of ERα and represent a promising strategy to tackle endocrine resistance. Here, we report a new class of SERDs by pharmacological evolution of a selective estrogen receptor modulator, lasofoxifene. The structure-activity relationship study and efforts to circumvent the issue of human ether-a-go-go-related gene led to the identification of compounds 51. This bifunctional compound displayed broad activity across a vast array of cell backgrounds and was capable of effectively degrading and antagonizing wild-type ERα and clinically relevant ERα mutants. 51 exhibited favorable pharmacokinetic properties and good brain penetration, with a brain/plasma ratio of 3.05, and significantly suppressed the growth of tumor in a tamoxifen-resistant MCF-7 Tam1 xenograft model. Overall, the study demonstrates 51 as a highly potent, oral, and brain penetrant ER degrader and pure antagonist, showing a good potential in overcoming endocrine resistance.

Antitumor effects of plasma‑activated solution on a murine melanoma model in vivo and in vitro.

Melanoma is a common malignant skin tumor with highly invasive features and a high metastasis rate that can be difficult to treat clinically. Large-scale resection of primary cutaneous melanoma is often used to avoid postoperative recurrence. For advanced patients, radiotherapy, targeted therapy and immunotherapy are often needed. Low-temperature plasma has been proved to have significant antitumor effects on a variety of cancer cell lines cultured in vitro. The main limitation of direct low-temperature plasma treatment is that it has weak penetration ability and can only treat superficial lesions. In recent years, research on low-temperature plasma-activated solution has revealed that it also have good antitumor effects and low-temperature plasma penetration depth problems can be solved by local injection. The present study revealed that low-temperature plasma-activated phosphate buffer solution exhibited good antitumor effects and biosafety against melanoma in vitro and in vivo. It demonstrated that low-temperature plasma-activated solution has antitumor effects due to its regulation of intracellular redox, destruction of mitochondrial function and DNA damage. In vivo experiments demonstrated that treatment with low-temperature plasma-activated solution not only exhibited antitumor effects but also caused no significant damage to hematopoietic function or liver and kidney functions in mice. All these results demonstrated that low-temperature plasma-activated solution represent a promising antitumor treatment strategy.

Physics-Based and Data Driven Modeling of Electrophoretic Coatings: Hybrid and Multiphysics Strategy

Electrophoretic coatings are used in several applications where surface treatment is intended to provide corrosion protection. These coatings are widely used in the automotive, aerospace, or many other sectors. These kind of coatings are electrochemically applied and have good levelling, penetration, adhesion, and corrosion resistance properties. A wide variety of materials can be deposited by this technique and among these coatings, this presentation will focus on ceramic deposits. In this case, electrophoretic deposition is carried out using a suspension containing ceramic particles and the deposition takes place on a conductive substrate through the application of an electric field. The control of process parameters and post-processing parameters (heat treatment or sintering) necessary to consolidate the raw coatings obtained can be difficult to define.In order to improve process management and understanding of electrophoretic deposition, we will explore several strategies and modelling theories implemented in the field of electrophoretic coating through a comprehensive analysis in order to optimize the coating process, improve quality and increase efficiency on complex geometry parts. Several theoretical approaches and models have been used to better understand electrochemical interactions and forces involved in electrophoresis to identify parameters related to electrophoretic mobility. The multiple parameters involved in electrophoretic coatings make the modelling of this process complex and the models developed are neither universal nor applicable to all chemistries which requires a specific development for each electrolyte and suspension.In a second part, we discuss limits of theoretical models and describes a strategy to develop a new modelling approach. This approach combines a physics-based model of current distribution and a data-driven optimization applied to electrophoretic coating. This methodology allows to model the thickness distribution on a part with complex geometry on an industrial scale, with the ability to adapt to a variation in the chemistry of the suspension used. This hybrid methodology allow to reduce experimental and fundamental characterizations, and can be easily applied to a new chemistry to estimate and well describe the throwing power at a macroscale.Furthermore, in some cases, physics-based modeling with secondary distribution can exhibit accuracy defects when the local concentration of particles drops, due to their consumption and low mass transport phenomena.In order to better describe the process behavior in this specific case, we develop a multiphysics model of tertiary current distribution than can account for the local distribution of particles, particularly near the part. This modeling approach allows to better anticipate thickness distribution in areas with low fluid mixing and agitation. The local efficiency is linked to the local current and particle concentration, as describe by a two phase flow model.The models developed are intended to help accurately predict and control the distribution of coating thickness and also local particle concentration to predict critical depletion zones.In conclusion, this conference explore different modeling strategies of electrophoretic coatings, by focusing on the optimization of process parameters and the consideration of the different mechanisms governing deposition with the objective of improving coating distribution on complex part.

Abstract C049: Implementing human bone marrow organoids to interrogate microenvironmental influences on the efficacy of blood cancer immunotherapies

Abstract Cancers perpetuate immunosuppressive niches that fuel progression and hinder the clinical utility of immunotherapies. Pre-clinical validation of immunotherapies often relies on 2D models that lack spatial complexity, or mouse models that do not fully recapitulate the human tumor microenvironment (TME). We recently developed a human induced pluripotent stem cell (iPSC)-derived bone marrow organoid platform, creating a robust system to interrogate cancer-stroma cross talk. Here, we further develop the model to create the first in vitro system for evaluation of blood cancer immunotherapies in a human tissue environment. We focused on targeting myeloproliferative neoplasms (MPNs) – chronic blood cancers affecting ∼400,000 individuals in the US that are currently largely incurable. 1/3 of MPNs are driven by a mutation in calreticulin (mutCALR), and the mutCALR oncoprotein is displayed on the cell surface of disease-initiating stem cells and fibrosis-driving megakaryocytes. MPNs are characterized by pronounced inflammation and fibrosis (“myelofibrosis”). Therapeutic antibodies have entered first-in-human trials, and we recently presented pre-clinical data for the first mutCALR-directed CAR-T cell therapy. However, it is unclear how the TME may limit the efficiency of these agents. To address this, we first tested the utility of the organoid platform to evaluate target killing of leukemic cell lines and cells from patients using the anti-CD33 antibody-drug conjugate (ADC) gemtuzumab. The ADC showed good penetration and potent target cell killing in organoids, with only minimal reduction in efficacy in TGFβ-rich/fibrotic microenvironments. We then developed more sophisticated ‘chimeroids’ comprising the iPSC organoid scaffold engrafted with malignant stem cells plus CAR-Ts. Addition of mutCALR-directed CAR-Ts induced highly selective killing of mutCALR+ stem/progenitor cells with minimal toxicity to non-malignant cells including niche cells (n=5 donors), replicating the highly specific and robust killing seen in 2D cultures. To evaluate the impact of TME perturbations on CAR-T phenotype and function, we generated a scRNAseq dataset of 161,970 cells from ‘healthy’ and ‘myelofibrotic’ organoids engrafted with patient-derived mutCALR+ cells plus CAR-Ts, pre-stimulating organoids with two immunoregulatory cytokines highly abundant in myelofibrosis -TGFβ and Galectin-1. CAR- Ts exposed to mutCALR+ cells showed a 2x expansion of CD8+ effector memory cells, with significant enrichment of IFNγ, TNFα and IL-2 signaling gene-sets. The proportion of immunosuppressive Tregs increased in a high TGFβ TME, but not with Galectin-1. Ongoing analyses are further interrogating crosstalk between CAR-T, target cells, and niche components, and exploring the pro-inflammatory impact of CAR-T killing on the marrow niche – a potential mediator of delayed hematopoietic recovery. These studies demonstrate the utility of human organoids to evaluate blood cancer-targeting immunotherapies within a relevant human TME, with broad relevance for mechanistic and translational studies. Citation Format: Zoë C. Wong, Yuqi Shen, Alex Rampotas, Isaac Gannon, Charlotte Brierley, Camelia Benlabiod, Nawshad Hayder, Eleanor Murphy, Aude-Anaïs Olijnik, Claire Roddie, Martin Pulé, Adam J. Mead, Abdullah O. Khan, Bethan Psaila. Implementing human bone marrow organoids to interrogate microenvironmental influences on the efficacy of blood cancer immunotherapies [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Tumor-body Interactions: The Roles of Micro- and Macroenvironment in Cancer; 2024 Nov 17-20; Boston, MA. Philadelphia (PA): AACR; Cancer Res 2024;84(22_Suppl):Abstract nr C049.

Pharmacokinetic Positron Emission Tomography Imaging of an Optimized CD38-Targeted 68Ga-Labeled Peptide in Multiple Myeloma: A Pilot Study.

Multiple myeloma (MM) is an incurable disease characterized by its clinical and prognostic heterogeneity. Despite conventional chemotherapy and autologous hematopoietic stem cell transplantation, the management of relapsed and refractory MM disease poses significant challenges, both medically and socioeconomically. CD38, highly expressed on the surface of MM cells, serves as a distinct tumor biological target in MM. Peptides offer advantages over antibodies, enabling precise tumor imaging and facilitating early tumor diagnosis and dynamic immunotherapy monitoring. In this study, we developed PF381, a CD38-targeted peptide, and investigated its role in diagnosis, biodistribution, and dosimetry through 68Ga-labeling for preclinical evaluation in tumor-bearing models. We screened a microchip-based combinatorial chemistry peptide library to obtain the amino acid sequence of PF381. Affinity for human CD38 was evaluated by SPRi. PF381 was conjugated with DOTA for radiolabeling with 68Ga, and the complex was characterized by HPLC. PET imaging was performed in murine tumor models after the administration of [68Ga]Ga-DOTA-PF381. Biodistribution analysis compared CD38-positive H929 and CD38-negative U266 tumors, and human radiation dosimetry was estimated. Tumor sections were stained for CD38 expression. SPRi showed that PF381 had a high affinity for CD38 with a KD of 2.49 × 10-8 M. HPLC measured a radiolabeling efficiency of 78.45 ± 7.91% for [68Ga]Ga-DOTA-PF381, with >98% radiochemical purity. PET imaging revealed rapid and persistent accumulation of radioactivity in CD38-positive H929 tumors, contrasting with negligible uptake in CD38-negative U266 tumors. Biodistribution confirmed higher uptake in H929 tumors (0.75 ± 0.03%ID/g) vs U266 (0.26 ± 0.08%ID/g, P < 0.001). The kidney received the highest radiation dose (3.57 × 10-02 mSv/MBq), with an effective dose of 1.41 × 10-02 mSv/MBq. Immunofluorescence imaging supported PET and biodistribution findings. We developed a novel peptide targeting CD38 and proved that 68Ga-labeled PF381 had rapid targeting and good tumor penetration capabilities. Therefore, 68Ga-labeled PF381 could achieve high sensitivity in vivo imaging for CD38-positive hematological malignancies.

The Neuroprotection of 1,2,4-Triazole Derivative by Inhibiting Inflammation and Protecting BBB Integrity in Acute Ischemic Stroke.

The oxidative stress and neuroinflammation are important factors in acute ischemic stroke (AIS). Our former study showed the 1,2,4- triazole derivative (SYS18) had obviously neuroprotection by anti- oxidative stress on rat middle cerebral artery occlusion (MCAO) model. In this study, we continue to investigate its neuroprotection by anti-inflammatory effects and protecting BBB integrity in AIS. First, its effect on acute inflammation was evaluated by the mice model of increased peritoneal capillary permeability. Then, the MCAO cerebral edema models were built to evaluate its neuroprotection by reducing the neurological score, cerebral edema, improving the biochemical indicators, and pathological damage of brain tissue. At the same time, its protection on blood-brain barrier (BBB) integrity was proved by decreasing the BBB permeability and inhibiting glycocalyx degradation and regulating the BBB tight junction proteins expression of matrix metalloproteinase- 9 (MMP- 9) and claudin- 5 in brain tissue. Meanwhile, pharmacokinetic experiments showed that the compound had good BBB penetration. It has some advantages in the intensity of efficacy compared with the marketed drug edaravone. Based on these findings, SYS18 has a strong potential to become a neuroprotectant in the future.

Synthesis, characterisation and functionalization of Hercynite nanoparticles for improved antibacterial activity

Hercynite i.e.,FeAl2O4 is an earth-abundant spinel mineral with a cubic crystal structure and belongs to the normal spinel ferrites possessing optical absorption in the visible range as well as superior magnetic and thermal properties. Herein, we synthesized nanosized FeAl2O4 where the citric acid-mediated sol–gel auto-combustion method was employed to achieve its pure phase and studied its physicochemical properties. Furthermore, the superior colloidal dispersion stability of the FeAl2O4 nanoparticles was achieved required for antibacterial activity and standardised via post-synthesis surface functionalisation using amino-propyl-triethoxysilane (APTES). We further characterised the material using states of art characterisation techniques for their structural, morphological, optical and thermal properties. Finally, the antibacterial activity of pure and surface functionalised FeAl2O4 nanoparticles was investigated against the Escherichia coli (E. coli) strain. We observed good penetration of surface functionalised FeAl2O4 nanoparticles into the bacterial membranes due to the high degree of dispersion achieved via cationic surface charge. Conclusively, a key finding of this study is the enhanced antibacterial properties of surface functionalised FeAl2O4 nanoparticles for the concentration of 31 µg/mL compared to pure FeAl2O4 nanoparticles at 62 µg/mL. This study has great relevance in the area of wound healing and tissue regeneration in the future.