Plasma Half-life Research Articles

The effects of breed and routes of administration on the plasma pharmacokinetics and faecal excretion of robenacoxib in goats

Robenacoxib (RX) is a non-steroidal anti-inflammatory drug (NSAID) of the coxib class. This study aimed to evaluate the plasma dispositions and faecal excretion profiles of RX in Alpine and Saanen goats following oral and subcutaneous routes. Two different goat breeds were allocated into two treatment groups concerning the breed. RX was administered subcutaneously to animals at a dose of 4 mg/kg b.w. Following a one-week washout period, RX was administered by oral route to the same animals at the same dose. Heparinized blood samples were collected from all animals before drug administration (0 h) and subsequently up to 24 h. Faecal samples were collected at various times between 8 h and 36 h. The concentrations of RX in plasma and faeces were determined by HPLC. The plasma half-life (T1/2λz) of RX in Saanen goats (1.21 h) was significantly longer (P < 0.017) than in Alpine goats (0.90 h) after subcutaneous administration. In both goat breeds, statistical differences were observed between subcutaneous and oral administration of RX for T1/2λz, Tlast, Cmax, AUC0-∞, and MRT0-∞. Faecal Cmax and Tmax parameters following oral administrations were 0.92 µg/g and 0.85 µg/g at 30 h and at 24 h in Alpine and Saanen goats, respectively. The difference in plasma protein ratio between Alpine and Saanen goats may have affected the T1/2λz of the drug. NSAIDs are among the drug groups frequently detected in aquatic and terrestrial ecosystems around the world and there are data on the effects of NSAID residues on wildlife and aquatic species. Therefore, revealing the excretion of NSAIDs, which are frequently used in the veterinary field, in faeces and urine should be considered for ecological sustainability.

Structure-based design, synthesis, and biological characterization of indolylarylsulfone derivatives as novel human immunodeficiency virus type 1 inhibitors with potent antiviral activities and favorable drug-like profiles.

In the current antiretroviral landscape, continuous efforts are still needed to search for novel chemotypes of human immunodeficiency virus type 1(HIV-1) inhibitors with improved drug resistance profiles and favorable drug-like properties. Herein, we report the design, synthesis, biological characterization, and druggability evaluation of a class of non-nucleoside reverse transcriptase inhibitors. Guided by the available crystallographic information, a series of novel indolylarylsulfone derivatives were rationally discovered via the substituent decorating strategy to fully explore the chemical space of the entrance channel. Among them, compound 11h bearing the cyano-substituted benzyl moiety proved to be the most effective inhibitor against HIV-1 wild-type and mutant strains (EC50 = 0.0039-0.338 μM), being far more potent than or comparable to etravirine and doravirine. Besides, 11h did not exhibit cytotoxicity at the maximum test concentration. Meanwhile, the binding target of 11h was further confirmed to be reverse transcriptase (IC50 = 0.055 μM). Preliminary structure-activity relationship were discussed to guide further optimization work. Molecular docking and dynamics simulation studies were investigated in detail to rationalize the biological evaluation results. Further drug-likeness assessment indicated that 11h possessed excellent physicochemical properties. Moreover, no apparent hERG blockade liability and cytochrome P450 inhibition were observed for 11h. Notably, 11h was characterized by favorable in vitro metabolic stability with moderate clearance rates and long half-lives in human plasma and liver microsomes. Overall, 11h holds great promise as an ideal Anti-HIV-1 lead compound due to its potent antiviral efficacy, low toxicity, and favorable drug-like profiles.

Development of a Clonal and High-Yield Mammalian Cell Line for the Manufacturing of a Hyperactive Human DNase I with Extended Plasma Half-Life Using PASylation® Technology.

Cumulative evidence from several pre-clinical studies suggests that restoration of plasma DNase activity in a thrombo-inflammatory state may improve clinical outcomes. Following injury, hyperactivated immune cells release large amounts of granular proteins together with DNA, which often accumulate in the surrounding environment in so-called neutrophil extracellular traps (NETs). Degradation of excess NETs by systemic DNase administration offers a promising therapeutic approach to ameliorate inflammation and dissolve intravascular clots. In order to expand the therapeutic utility of human DNase I, a variant of the enzyme was developed that has both a prolonged systemic half-life and a higher catalytic activity compared to Dornase alfa (Pulmozyme®), the recombinant form of DNase I approved for inhaled therapy of cystic fibrosis. The hyperactive enzyme was "PASylated" by genetic fusion with a strongly hydrophilic and biodegradable PAS-polypeptide to increase its hydrodynamic volume and retard kidney filtration. A stable TurboCell™ CHO-K1-based cell line was generated which is suitable for the future production of PASylated DNase I according to good manufacturing practice (GMP). Furthermore, a robust bioprocess strategy was devised and an effective downstream process was developed. The final protein product is characterized by excellent purity, favorable physicochemical properties, a 14-fold higher DNA-degrading activity than Dornase alfa and a sustained pharmacokinetic profile, with a 22-fold slower clearance in rats.

Molecular dynamics-guided optimization of BGM0504 enhances dual-target agonism for combating diabetes and obesity

The dual activation of glucagon-like peptide-1 receptor (GLP-1R) and glucose-dependent insulinotropic polypeptide receptor (GIPR) has emerged as a promising therapeutic strategy for managing type 2 diabetes and obesity. Tirzepatide, a dual agonist peptide, has exhibited superior clinical efficacy in glycemic and weight control compared to selective GLP-1R agonists. Nevertheless, the structural basis of Tirzepatide's extended half-life, attributed to an acylation side chain on the parent peptide, raises questions regarding its partial agonistic activity. Employing molecular dynamics simulations, we explored the dynamic processes of peptide-receptor interactions. We uncovered a crucial salt bridge between parent peptide and GLP-1R/GIPR at K20, a feature not discernible in cryo-electron microscopy structures. Building upon these insights, we developed an optimization strategy based on the parent peptide which involved repositioning the acylation side chain. The results of both in vitro and in vivo experiments demonstrated that the optimized peptide has twofold to threefold increase in agonistic activity compared to Tirzepatide while maintaining its extended half-life in plasma. This led to the design of BGM0504, which proved to be more effective than its predecessor, Tirzepatide, in both laboratory and animal studies.

Characterization of recombinant factor IX fusion proteins enabling subcutaneous administration

BackgroundThe X-linked bleeding disorder hemophilia B, caused by mutation(s) in the coagulation factor (F)IX gene, leads to partial or total loss of its function, requiring lifelong FIX replacement therapy. Although new recombinant FIX (rIX) therapeutics like albumin fusion proteins (rIX-FP) enable longer plasma half-life and thus less frequent administration, the complexity of intravenous (i.v.) injection remains. ObjectivesThe study aimed to characterize rIX-FP variants with anticipated enhanced specific activity, which would leverage rIX-FP’s superior pharmacokinetic profile with beneficial characteristics for subcutaneous (s.c.) administration. MethodsTwo rIX-FP variants, R338L (“Padua variant”) and R338L/E410K, were characterized in vitro. Pharmacokinetic profiles of FIX antigen and activity levels were evaluated in FIX-deficient mice after i.v. and s.c. administration of these variants (dosing based on antigen levels). The efficacy of the most promising variant was tested after i.v. and s.c. administration (dosing based on activity) in a tail clip bleeding model. A marketed wild-type (WT) rIX-FP product served as the comparator. ResultsBoth rIX-FP variants showed a 4- to 5-fold increase in specific activity in vitro compared with rIX(WT)-FP, while FXIa-mediated activation was the fastest for rIX(WT)-FP and rIX(R338L)-FP. Compared with rIX(WT)-FP and rIX(R338L/E410K)-FP, rIX(R338L)-FP exhibited higher FIX activity exposure after i.v. and s.c. administration and demonstrated comparable efficacy with rIX(WT)-FP in reducing bleeding time and blood loss in FIX-deficient mice requiring ∼4 times lower protein amount. ConclusionrIX(R338L)-FP was shown to be a promising candidate for s.c. administration, exhibiting increased specific activity combined with higher activity-based exposure and indicating efficacy at a lower protein dose.

Genetically engineered long-acting Esculentin-2CHa(1−30) fusion protein with potential applicability for the treatment of NAFLD

Esculentin-2CHa(1–30) (‟ESC”) has been reported as a potent anti-diabetic peptide with little toxicity. However, its very short plasma residence time severely limits the therapeutic efficacy. To address this issue, we genetically engineered a fusion protein of tandem trimeric ESC with an albumin binding domain (ABD) and a fusion partner, SUMO (named ‟SUMO-3×ESC-ABD”). The SUMO-3×ESC-ABD, successfully produced from E. coli, showed low cellular and hemolytic toxicity while displaying potent activities for the amelioration of hyperglycemia as well as non-alcoholic fatty liver disease (NAFLD) in vitro. In animal studies, the estimated plasma half-life of SUMO-3×ESC-ABD was markedly longer (427-fold) than that of the ESC peptide. In virtue of the extended plasma residence, the SUMO-3×ESC-ABD could produce significant anti-hyperglycemic effects that lasted for >2 days, while both the ESC or ESC-ABD peptides elicited little effects. Further, twice-weekly treatment for 10 weeks, the SUMO-3×ESC-ABD displayed significant improvement in blood glucose control with a reduction in body weight. Most importantly, a significant improvement in the conditions of NAFLD was observed in the SUMO-3×ESC-ABD-treated mice. Along the systemic effects (by improved glucose tolerance and body weight reduction), direct inhibition of the hepatocyte lipid uptake was suggested as the major mechanism of the anti-NAFLD effects. Overall, this study demonstrated the utility of the long-acting SUMO-3×ESC-ABD as a potent drug candidate for the treatment of NAFLD.

A phase I randomized study to evaluate safety, pharmacokinetics, and pharmacodynamics of SIR2446M, a selective RIPK1 inhibitor, in healthy participants.

Activation of receptor-interacting protein kinase 1 (RIPK1), a broadly expressed serine/threonine protein kinase, by pro-inflammatory cytokines and pathogens can result in apoptosis, necroptosis, or inflammation. RIPK1 inhibition has been shown to reduce inflammation and cell damage in preclinical studies and may have therapeutic potential for degenerative and inflammatory diseases. SIR2446 is a potent and selective novel small molecule RIPK1 kinase inhibitor. This phase I, randomized, double-blind, placebo-controlled study in Australia (ACTRN12621001621808) evaluated the safety (primary objective), pharmacokinetics, and pharmacodynamics of single (3-600 mg) and multiple (5-400 mg for 10 days) ascending oral doses of SIR2446M (SIR2446 magnesium salt form) in healthy adults from Nov 24, 2021, until May 01, 2023. All treatment-emergent adverse events (TEAEs) were mild/moderate. The most reported TEAEs were vascular access site pain, headache, and rash morbilliform. SIR2446M plasma half-lives ranged from 11 to 19 h and there were no major deviations from dose proportionality for maximum concentration and area under the curve across doses. Renal excretion of unchanged SIR2446 was minimal. No marked accumulation was observed (mean accumulation ratio, 1.2-1.6) after multiple daily doses. A high-fat meal mildly reduced the exposure but was not considered clinically significant. SIR2446M had a rapid and sustained inhibitory effect on the activity of RIPK1, with an overall 90% target engagement at repeated doses ranging from 30 to 400 mg in peripheral blood mononuclear cells exvivo stimulated to undergo necroptosis. The favorable safety, pharmacokinetic, and pharmacodynamic profile of SIR2446M in healthy participants supports its further clinical development in patients with degenerative and inflammatory diseases.

Improving Circulation Half-Life of Therapeutic Candidate N-TIMP2 by Unfolded Peptide Extension.

Matrix Metalloproteinases (MMPs) are drivers of many diseases including cancer and are established targets for drug development. Tissue inhibitors of metalloproteinases (TIMPs) are human proteins that inhibit MMPs and are being pursued for the development of anti-MMP therapeutics. TIMPs possess many attractive properties of a drug candidate, such as complete MMP inhibition, low toxicity and immunogenicity, high tissue permeability and others. A major challenge with TIMPs, however, is their formulation and delivery, as these proteins are quickly cleared from the bloodstream due to their small size. In this study, we explore a new method for plasma half-life extension for the N-terminal domain of TIMP2 (N-TIMP2) through appending it with a long intrinsically unfolded tail containing a random combination of Pro, Ala, and Thr (PATylation). We design, produce and explore two PATylated N-TIMP2 constructs with a tail length of 100- and 200-amino acids (N-TIMP2-PAT100 and N-TIMP2-PAT200, respectively). We demonstrate that both PATylated N-TIMP2 constructs possess apparent higher molecular weights compared to the wild-type protein and retain high inhibitory activity against MMP-9. Furthermore, when injected into mice, N-TIMP2-PAT200 exhibited a significant increase in plasma half-life compared to the non-PATylated variant, enhancing the therapeutic potential of the protein. Thus, we establish that PATylation could be successfully applied to TIMP-based therapeutics and offers distinct advantages as an approach for half-life extension, such as fully genetic encoding of the gene construct, mono-dispersion, and biodegradability. Furthermore, PATylation could be easily applied to N-TIMP2 variants engineered to possess high affinity and selectivity toward individual MMP family members, thus creating attractive candidates for drug development against MMP-related diseases.

TRLS-17. INTERIM CLINICAL, PHARMACOKINETIC, AND TECHNICAL REPORT OF A PHASE 2 STUDY OF SONODYNAMIC THERAPY (SDT) USING SONALA-001 TOGETHER WITH MR-GUIDED LOW-INTENSITY FOCUSED ULTRASOUND (MRGFUS) IN CHILDREN WITH DIFFUSE INTRINSIC PONTINE GLIOMA (DIPG)

Abstract BACKGROUND Children with DIPG have limited treatment options and poor prognosis with median survival of 9-12 months. Preclinical studies reveal that sonodynamic therapy (SDT) via MR-guided focused ultrasound (MRgFUS) activates SONALA-001 (ALA) metabolite, protoporphyrin IX (PpIX), inducing tumor cell death and extending survival in glioma models. METHODS We present preliminary data from an ongoing first-in-child drug-device trial (NCT05123534) of SONALA-001 SDT in children with DIPG aged ≥ 5 years post-radiation therapy. The trial evaluates the safety of ALA-SDT, preliminary efficacy and pharmacokinetics (PK) of SONALA-001 at 5mg/kg administered 6-12 hours before SDT and establishes the RP2D. Each cohort involves three subjects: the first receiving half-pons treatment 30-days apart; the others receiving entire pons treatment. Single-treatment approach was amended to allow up to 12 monthly-treatments. RESULTS Six patients (4M:2F, 5-12 years) in 2 cohorts received 22 SDT-treatments between August 2022 and February 2023. Sonication duration averaged 147 minutes. Five of 6 patients received repeated treatments. All were discharged post-procedure day 1 with no DLTs or related AE grade ≥ 3. For SONALA-001, the Cmax occurred at end of infusion, followed by rapid clearance (15.7 mL/min/kg); plasma half-life of &amp;lt; 1 hour, indicating rapid distribution. The PpIX-Cmax occurred 4-6 hours post-dose, declining with a longer mean half-life than SONALA-001 of 6 hours. Both Cmax and AUCall demonstrate circulating-PpIX and systemic exposure were significantly lower than for SONALA-001. Two of 6 patients achieved a partial response per RAPNO central-review, and 2 continue in the study treatment for 11 and 15 months, respectively. CONCLUSION SDT is an innovative and to-date well-tolerated strategy for children with DIPG post-initial radiotherapy. Sonications did not result in any adverse procedural effects. SONALA-001 exhibits rapid distribution and clearance, while PpIX PK align with metabolite formation. After dose escalation completion, dose-expansion at R2PD will offer insights into safety, PK, and efficacy.

Wound-healing effects of curcumin and its nano formulation

Wound healing is a method of tissue repair or regeneration. Section Damages. Plants and plant-derived bioactive substances have been found effective in the treatment of many diseases. Types of wounds. Curcumin is a natural polyphenol that has been used since ancient times. Ayurveda is used times for its healing properties to relieve pain and aid in many healing processes. Plates. Various studies of curcumin administration to the site of pain have reported the following effects: Curcumin eliminates reactive oxygen species and increases the ability to improve collagen deposition, Chapter: Granulation data finally makes the wound contract. Curcumin is widely available and has been studied for its ability to relieve pain, but in addition it has low solubility and rapid metabolism. The short plasma half-life limits its application in wound healing. Since nanotechnology existed Chapter Proven to be a good method for wound healing with proper support Chapter injury sites. This review highlights the potential of curcumin and its Nano formulations; Preparation of liposomes, nanoparticles, nano emulsions, etc. usage This article is about Curcumin’s many biomedical applications underlie its anti-biofilm properties Chapter and its wound healing effect.

Disposition kinetics and tissue residues of tilmicosin following intravenous, subcutaneous, single and multiple oral dosing in geese (Anser anser domesticus).

Tilmicosin (TMC), a semi-synthetic macrolide antibiotic, is widely used in veterinary medicine due to its broad-spectrum, bacteriostatic properties. Frequently administered in various birds species, it is likely used off-label in geese as well. The study sought to investigate TMC's pharmacokinetics, tissue residues, in geese through invivo experiments. The study involved longitudinal open studies on 15 healthy adult males, with three phases separated by one-month washout periods. Geese were administered TMC through intravenous (IV, 5 mg/kg), subcutaneous (SC, 10 mg/kg), and oral (PO, 25 mg/kg for five consecutive days) routes, with blood samples drawn at specific intervals. Tissue samples were also collected for subsequent analysis at pre-assigned times. TMC in goose plasma was quantified by a fully validated HPLC method. Plasma concentrations were quantified up to 4 hr for the PO and IV routes, and up to 10 hr in the SC route. Significant variations in bioavailability were observed between SC (87%) and PO (4%) routes. The body extraction ratio was low at 0.03, suggesting minimal ability of the liver and kidneys to eliminate TMC. Multiple oral doses showed no plasma accumulation, but tissue data revealed extensive distribution and prolonged residence, up to 120 h, suggesting a sustained therapeutic effect despite the brief plasma half-life. Regarding the multiple PO doses, provisional withdrawal times of 6, 7.5, and 8 days were suggested for the liver, muscles, and kidneys, respectively, according to the MRL set for these matrices in chickens by EMA. In conclusion, while the practical oral administration is discouraged at the population level, SC administration of TMC may be appropriate for geese, albeit impractical for flock therapy.

Revolutionizing Stroke Care: Nanotechnology-Based Brain Delivery as a Novel Paradigm for Treatment and Diagnosis.

Stroke, a severe medical condition arising from abnormalities in the coagulation-fibrinolysis cycle and metabolic processes, results in brain cell impairment and injury due to blood flow obstruction within the brain. Prompt and efficient therapeutic approaches are imperative to control and preserve brain functions. Conventional stroke medications, including fibrinolytic agents, play a crucial role in facilitating reperfusion to the ischemic brain. However, their clinical efficacy is hampered by short plasma half-lives, limited brain tissue distribution attributed to the blood-brain barrier (BBB), and lack of targeted drug delivery to the ischemic region. To address these challenges, diverse nanomedicine strategies, such as vesicular systems, polymeric nanoparticles, dendrimers, exosomes, inorganic nanoparticles, and biomimetic nanoparticles, have emerged. These platforms enhance drug pharmacokinetics by facilitating targeted drug accumulation at the ischemic site. By leveraging nanocarriers, engineered drug delivery systems hold the potential to overcome challenges associated with conventional stroke medications. This comprehensive review explores the pathophysiological mechanism underlying stroke and BBB disruption in stroke. Additionally, this review investigates the utilization of nanocarriers for current therapeutic and diagnostic interventions in stroke management. By addressing these aspects, the review aims to provide insight into potential strategies for improving stroke treatment and diagnosis through a nanomedicine approach.

Anticoagulation in Pediatric Patients

The use of antithrombotic agents is increasing in infants, children and adolescents. The more recent routine inclusion of children in FDA-monitored clinical trials has propelled the rapid accumulation of safety and efficacy data on these agents in pediatric patients. Antithrombotic agents in current use include indirect or antithrombin-dependent anticoagulants, intravenous direct thrombin inhibitors, direct oral anticoagulants (DOACs) targeting thrombin or factor Xa, antiplatelet agents and thrombolytic therapies. Each class of antithrombotic agent has distinct mechanisms of action, clearance routes, half-lives, safety and dosing. Anticoagulant efficacy is dependent upon the specific clinical indication and stability of the pediatric patient. Duration of anticoagulant course is also dependent upon the clinical indication as well as rate of thrombus resolution. This manuscript reviews the mechanism of action, route of administration, route of clearance and plasma half-life for the antithrombotic agents in current use in children. Use of anticoagulation in the context of thrombolytic therapy is discussed.

Daily compared with alternate-day levamisole in pediatric nephrotic syndrome: an open-label randomized controlled study.

Levamisole is less expensive and has a better toxicity profile compared to other steroid sparing agents used in nephrotic syndrome. It has a plasma half-life of 2.0 to 5.6hours, but is conventionally administered on alternate days. We aimed to assess whether daily levamisole is safe and more effective than standard alternate-day therapy in maintaining remission in children with frequently relapsing or steroid-dependent nephrotic syndrome (FR/SDNS). An open-label randomized controlled trial was conducted in children with FR/SDNS. Group A received daily while Group B received alternate-day levamisole (2-3mg/kg/dose) for 12months. Prednisolone was tapered off by 3months. Patients were monitored for relapses, further steroid requirement, and adverse effects. A total of 190 children with FR/SDNS (94 in Group A and 96 in Group B) were analyzed. Sustained remission for 12months was observed in 36% of Group A and 27% of Group B patients (p = 0.18). Numbers completing 12months in the study were 67% in Group A and 56% in Group B (p = 0.13). Time to first relapse, persistent FR/SDNS, and withdrawal due to poor compliance were statistically similar in both groups, while relapse rate and cumulative steroid dosage were significantly lower in Group A compared to Group B (p = 0.03 and p = 0.02, respectively). The incidence of adverse effects was comparable in both groups, with reversible leucopenia and hepatic transaminitis being the commonest. Daily levamisole therapy was not superior to alternate-day therapy in maintaining sustained remission over 12months. Nevertheless, relapse rate and cumulative steroid dosage were significantly lower without increased adverse effects.

Interim report of a phase 2 study of sonodynamic therapy (SDT) using SONALA-001 together with MR-guided low-intensity focused ultrasound (MRgFUS) in children with diffuse intrinsic pontine glioma (DIPG).

2067 Background: Children with DIPG have limited treatment options and a poor prognosis, with median survival of 9-12 months. Preclinical studies reveal that sonodynamic therapy (SDT) with MR-guided focused ultrasound (MRgFUS) activates ALA metabolite, protoporphyrin IX (PpIX), leading to tumor cell death and extending survival in glioma models. Methods: SDT-201 (NCT05123534) is a multi-center, first-in-child dose escalation trial of SDT with SONALA-001 and MRgFUS administered in children aged ≥ 5 years with DIPG, enrolled 4-20 weeks post-radiation therapy and prior to disease progression. The trial evaluates the safety and preliminary efficacy of SDT and pharmacokinetics (PK) of intravenous ALA (SONALA-001) at 5mg/kg, administered 6-12 hours before MRgFUS to the pons (delivered to half the pons 30 days apart for the first patient in each cohort of 3, with subsequent patients receiving entire pons treatment). The trial was subsequently amended to allow up to 12 monthly treatments. Results: Six patients in 2 cohorts were treated (4 male:2 female, range 5-12 years) between August 2022 and December 2023. No DLTs or related AE grade ≥ 3 were observed. For SONALA-001, the Cmax occurred at the end of infusion, followed by rapid clearance (15.7 mL/min/kg) with plasma half-life of &lt; 1 hour, indicating rapid distribution. The PpIX Cmax occurred 4-6 hours post-dose and declined with a longer mean half-life than SONALA-001 of 6 hours. Both Cmax and AUCall demonstrate circulating-PpIX and systemic exposure were significantly lower than for SONALA-001. Two of the first 6 patients achieved a partial response per RAPNO central-review, and 2 subjects continuing treatment are 11 to 15 months from start of study therapy. Conclusions: SDT is a well-tolerated strategy for patients with DIPG after initial radiotherapy. SONALA-001 shows rapid distribution and clearance, and PpIX PK align with metabolite formation. Following dose escalation completion, dose-expansion will be initiated at RP2D offering further insights into safety, PK, and efficacy. Clinical trial information: NCT05123534 .

Efficacy and biomarker analysis of phase 2 (P2) and window-of-opportunity (WoO) cohorts of patients with recurrent glioblastoma (rGBM) treated with ST101, an inhibitor of the transcription factor C/EBPβ.

2011 Background: CCAAT/enhancer-binding protein β (C/EBPβ) is an important transcription factor in GBM cell maintenance and is associated with worse outcomes and resistance to chemotherapy through transcription of genes associated with cell cycle and DNA repair. C/EBPβ is also required for maintenance of immunosuppressive tumor-associated macrophages. ST101 is a first-in-class antagonist of C/EBPβ that promotes selective GBM cell death without impacting normal cell viability (Darvishi, 2022), and repolarization of macrophages toward an immunostimulatory program. In an all comers P1 study, ST101 was well-tolerated, showed significant tumor uptake, ~40 hr plasma half-life, and signal of efficacy (1PR +8SD). With strong rationale targeting C/EBPβ in GBM and evidence that ST101 crosses the BBB in preclinical biodistribution studies, cohorts in rGBM were explored. Methods: Pts with ≤2 recurrences of GBM were enrolled in a P2 cohort (n=30) and treated with 500mg QW. A WoO cohort of 6 pts with rGBM received 2-4 doses of ST101 500mg QW prior to surgery and resumed ST101 after surgery until progression (NCT04478279). Endpoints for the two cohorts included response, PFS and OS while the WoO cohort also included tissue analyses (GBM subtypes, tumor microenvironment (TME) characterization, ST101 tumor penetration, and C/EBPβ target engagement). Clinical data is available for both cohorts and tissue analysis is available for the WoO cohort. Results: Data is a snapshot as of February 1, 2024. ST101 was safe and well-tolerated in both cohorts with no discontinuations due to AEs. Pts in the P2 study show 30% DCR (1PR + 8SD), 20% 6-mo PFS and 37% OS at 1 year. Median duration of treatment is 3.4 mos with 2 pts stable and still on study for 10+ and 16+ mos. In the WoO study, efficacy from post-surgery MRI showed 67% DCR (2 PR + 2 SD) and ~50% 6-mo PFS. Median OS cannot be assessed at this early stage. The median duration of treatment is 6.02 mos with 2 pts still on study for 7+ and 10+ mos. Tissue compared before and after ST101 exposure shows ST101 uptake and C/EBPβ target engagement by IHC, and pathologic evidence of treatment effect, such as geographic necrosis, in 5/6 pts. Evaluation of tumor and TME characteristics in post-ST101 treatment tissue will be presented. Conclusions: Monotherapy ST101 in pts with rGBM has an excellent safety profile and results in comparable outcomes to current 2nd line treatments. ST101 crosses the BBB, engages its target, and potentially induces treatment-related necrosis. These encouraging clinical activity, safety, and tissue findings, as well as preclinical data showing additive activity in combination with lomustine, support advancing ST101 to a randomized, placebo-controlled study comparing ST101 + lomustine vs. lomustine + placebo in rGBM. We aim to initiate this study in Q3 2024. Clinical trial information: NCT04478279 .

Study update of the oral CDK9 inhibitor KB-0742 in relapsed or refractory transcriptionally addicted advanced solid tumors.

3102 Background: KB-0742 is a potent, and selective, oral inhibitor of CDK9 being evaluated in a phase I/II study in patients with transcriptionally addicted advanced solid tumors (NCT04718675). Interim data from the first 4 dose levels were presented previously noting manageable safety (MTD not reached), a 24-hour plasma half-life, linear PK, CDK9 target engagement in peripheral blood mononuclear cells (PBMCs), and anti-tumor activity in patients with transcription factor fusion (TFF) driven sarcomas. Here we present updated KB-0742 safety, pharmacokinetics (PK), pharmacodynamic data (PD) and anti-tumor activity for patients from the ongoing dose escalation through 5 dose levels and 60 mg expansion. Methods: Study objectives include evaluation of safety, tolerability, PK, PD, and identification of KB-0742 MTD and RP2D. KB-0742 is administered orally once daily for 3 consecutive days followed by 4 days off, weekly in 28-day cycles, until unacceptable toxicity or disease progression. Eligible patients were enrolled in 5 escalation cohorts (10, 20, 40, 60 and 80 mg) or 60 mg dose expansion. Eligibility criteria include age &gt;18 years, relapsed or refractory solid tumors, and ECOG PS &lt; 2. PD is assessed in (PBMCs) and tumor tissue from pre- and on-treatment biopsy samples. Results: As of January 4, 2024, 112 patients were enrolled, 42 in dose escalation and 70 in expansion. Patients received a median of 3 lines of prior therapy. The most common tumor types enrolled were soft tissue sarcoma (STS) (n=36; 18 TFF positive) and adenoid cystic carcinoma (ACC) (n=18). Treatment-emergent adverse events occurring in &gt;15% of patients include nausea, vomiting, anemia, fatigue, diarrhea, and constipation; none assessed as grade 4 or 5. The most common reason for treatment discontinuation was disease progression (54.5%). Across 5 dose levels, PK remains linear with a terminal half-life of 24 hours. At 60 mg, evidence of target engagement was observed in post-treatment paired tumor tissue biopsies. Within STS, TFF positive patients displayed a trend towards improved outcomes vs. those without a TFF with a disease control rate (DCR) of 42.8% vs. 29.4%, and one partial response was observed in a patient with TFF positive myxoid liposarcoma at 60mg. The best observed response was durable stable disease (SD) yielding a DCR of 53.8% in ACC (n=18), and 83% in NSCLC (n=6). Two patients (MYCL1+ ovarian, NSCLC) with prolonged SD (&gt;140 days) continue treatment on 60mg. Conclusions: KB-0742 treatment at 60 and 80 mg was well tolerated, with manageable toxicity. Achievement of long-term SD and some preliminary anti-tumor efficacy in highly pretreated patients motivates continued enrollment of patients with transcriptionally addicted tumors. Dose escalation and expansion in transcriptionally addicted ( MYC amplification/overexpression) or TFF driven tumors continues. Clinical trial information: NCT04718675 .

A phase 2/3 study of Bicycle toxin conjugate BT8009 targeting nectin-4 in patients with locally advanced or metastatic urothelial cancer (la/mUC): Duravelo-2.

TPS4619 Background: BT8009 is a Bicycle toxin conjugate (BTC), comprising a highly selective bicyclic peptide targeting nectin-4 linked to the cytotoxin monomethyl auristatin E (MMAE) via a cleavable linker. Nectin-4 is an adhesion molecule commonly expressed in many tumor types, including urothelial cancer (UC), and is a validated therapeutic target (Hoffman-Censits 2021). BT8009 has a low molecular weight and short plasma half-life, with potential to rapidly penetrate solid tumors and reduce toxicity by minimizing exposure to normal tissue (Rigby 2022). Results from the ongoing phase 1/2 clinical trial of BT8009 (NCT04561362) indicate preliminary antitumor activity and a tolerable safety profile in patients (pts) with advanced malignancies including UC (Baldini 2023). This global, open label, phase 2/3 multicenter adaptive study aims to evaluate the safety and efficacy of BT8009 as monotherapy, or combined with pembrolizumab (pembro), vs chemotherapy in pts with locally advanced or metastatic UC (la/mUC) (NCT06225596/BT8009-230; Duravelo-2). Methods: The trial will enroll up to 956 adult pts in 2 cohorts. Cohort 1 will include n≤641 previously untreated pts eligible for platinum-based chemotherapy. Cohort 2 will include n≤315 pts with ≥1 prior systemic therapy, excluding enfortumab vedotin or other MMAE-based therapy. Pts must have la/mUC of the renal pelvis, ureter, bladder, or urethra, ECOG performance status ≤2 (Cohort 1) or ≤1 (Cohort 2), and adequate organ function. Cohort 1 will be randomized 1:1:1 to receive: 1) BT8009 5 mg/m2 on days (D)1, 8, and 15 + pembro 200 mg on D1; 2) BT8009 6 mg/m2 on D1 and 8 + pembro 200 mg on D1; or 3) chemotherapy (gemcitabine + cisplatin / carboplatin, followed by avelumab maintenance). Cohort 2 will be randomized 1:1 to receive: 1) BT8009 5 mg/m2 on D1, 8, and 15 or 2) BT8009 6 mg/m2 on D1 and 8. Cycle lengths will be 21D (28D for avelumab). After 30 pts in each dose arm have 9 weeks follow-up, an interim analysis will determine the optimal dose of BT8009 + pembro (Cohort 1) or BT8009 monotherapy (Cohort 2) to be used for the rest of the study. An additional Cohort 2 arm, optimal dose of BT8009 + pembro, will open after completion of the interim analysis. Discontinuation criteria include planned completion of therapy, progressive disease, and intolerable toxicity. Primary endpoints are progression-free survival (PFS; Cohort 1) and objective response rate (ORR; Cohort 2) assessed by blinded independent central review. Secondary endpoints are ORR (Cohort 1), PFS (Cohort 2), and overall survival, duration of response, disease control rate, safety/tolerability, and health-related quality of life (Cohorts 1 and 2). Pharmacokinetics, incidence/titers of antidrug antibodies, and tumor/peripheral biomarkers are exploratory endpoints. Efficacy endpoints will be assessed per RECIST v1.1. This study is actively recruiting. Clinical trial information: NCT06225596 .

#753 Targeted anti-IL-1β bacteroides fragilis outer membrane vesicles alleviate inflammatory injury in diabetic nephropathy

Abstract Background and Aims It is accepted that diabetic nephropathy (DN) is not only a disease caused by impaired glucose metabolism, but also a chronic inflammatory disease. The interleukin-1β (IL-1β) plays an important role in inflammation-mediated renal injury in DN. IL-1β blocking therapy can prevent inflammatory cytokine-mediated kidney injury. However, the high cost and systemic side effects of IL-1β antibodies limit its application. Therefore, firstly this study aimed to construct kidney-targeting outer membrane vesicles (OMVs) of Bacteroides fragilis loaded with IL-1β single-chain variable fragment (scFv). These OMVs derived from human commensal non-toxigenic Bacteroides fragilis have been proven to play an immunosuppressive role in the intestine and distant organs. This targeted drug delivery platform built with this as a carrier is expected to relieve kidney inflammation in the development of DN. Methods We recombinantly expressed the scFv based on the Gevokizumab sequence in Escherichia coli, and loaded it into the OMVs derived from Bacteroides fragilis by multiple ultrasounds. The targeting peptide with the sequence (KKEEE)3{Lys(N3)} was connected to the membrane surface through a copper-free catalyzed click chemistry reaction. The package, which consisted of OMVs and the scFv targeting IL-1β in the kidney (OMV-KTP-scFv) was obtained. OMV-KTP-scFv treated in HK-2 cells and healthy mice respectively, and evaluated its in vitro and in vivo drug safety. The concentration of IL-1β scFv in blood and kidney tissues at different time points were measured using streptozotocin (STZ)-induced diabetic mice. DID-labeled OMVs was used for ex vivo infrared imaging of various organs to evaluate the delivery efficiency of the targeted carrier. In vitro, OMV-KTP-scFv was used to prevent inflammation in mouse primary renal tubular epithelial cells induced by high glucose, and in vivo to alleviate kidney injury in STZ-induced diabetic mice. Results We successfully prepared OMV-KTP-scFv and confirmed the co-localization of OMVs, KTP, and scFv using super-resolution microscopy. The prepared OMV-KTP-scFv had high stability, effectively extended the plasma half-life of scFv, and could deliver scFv accurately to the proximal renal tubules. Furthermore, OMV-KTP-scFv also had good safety in vitro and in vivo, and no obvious biotoxicity was found during long-term in vivo administration. In addition, our results show that OMV-KTP-scFv significantly inhibited the expression of inflammatory factors in renal tubular cells of DN, reduced the infiltration of renal interstitial inflammatory cells, and alleviated renal tubular injury. Conclusion Our findings demonstrated that OMVs of Bacteroides fragilis targeting the kidney can deliver the IL-1β scFv to the renal tubulointerstitium and reduce the renal interstitial injury in DN by antagonizing local inflammation.

Small extracellular vesicle-based delivery of interleukin-10 improves treatment of experimental autoimmune uveitis

Non-infectious uveitis is an intraocular autoimmune disease mainly characterized by immune dysregulation of the eye, which may cause blindness if not well treated. Interleukin 10 (IL-10) is a potent cytokine with multiple immunoregulatory functions. However, it's in vivo activity is unstable owing to its inherent protein instability and short plasma half-life. Therefore, our previous research tried to establish IL-10-overexpressing MSC-sEVs (sEVs-IL10) using lentiviral transfection. While this approach indeed improved drug delivery, it also suffered from tedious procedures and limited loading efficiency. Accordingly, we constructed a novel MSC-sEVs-based delivery system for IL-10 (IL-10@sEVs) by sonication. The obtained formulation (IL-10@sEVs) had relatively higher loading efficiency and exerted a more profound immunomodulatory effect than sEVs-IL10 in vitro. Furthermore, IL-10@sEVs had significant therapeutic effects in a mouse model of experimental autoimmune uveitis (EAU) by decreasing the percentage of Th17 cells, increasing regulatory T cells in the eye, and draining lymph nodes. In summary, sonication outperforms conventional transfection methods for loading IL-10 into MSC-sEVs.