Subcutaneous Dose Research Articles

Mim8, a novel factor VIIIa mimetic bispecific antibody, shows favorable safety and pharmacokinetics in healthy adults

BackgroundMim8 (denecimig) is a novel activated coagulation factor VIII-mimetic bispecific antibody that assembles with activated coagulation FIX and FX on the platelet membrane surface. ObjectivesThe FRONTIER1 (NCT04204408, NN7769-4513) single ascending dose and the 4882 pharmacokinetic (PK) studies (NCT05127473, NN7769-4882) examined the safety, tolerability, PK, and pharmacodynamics (PD) of Mim8 in healthy adult males. MethodsThe FRONTIER1 single ascending dose study consisted of 6 cohorts, each with 6 participants who received a single subcutaneous (s.c.) dose of Mim8 and 2 participants who received a placebo. The 4882 PK study had 11 arms, each with 6 participants who received a single s.c. dose of Mim8. The primary endpoint for both studies was treatment-emergent adverse events. Other safety assessments included relative changes in D-dimer, prothrombin fragments 1 and 2, fibrinogen, and platelets. The PK and PD were assessed using Mim8 plasma concentration and activated partial thromboplastin clotting time and thrombin generation, respectively. ResultsMim8 was well tolerated, and there were no severe treatment-emergent adverse events. The PK properties of Mim8 in both studies were consistent with dose-proportionality. The terminal half-life of Mim8 after a single dose was approximately 1 month, and maximum plasma concentration was reached after 10 days. ConclusionThe PK and PD profiles suggest that Mim8 is suitable as a long-acting FVIIIa-mimetic bispecific antibody for hemophilia A prophylaxis.

Hypervitaminosis D induced hypercalcemia leading to coma: A case report

An elderly female presented with abdominal pain, vomiting and easy fatigability. Her sensorium gradually declined and became comatose. She was on Methotrexate, folic acid and prednisolone for her rheumatoid arthritis. The neurological examination and neuroimaging did not contribute in making the diagnosis. It is challenging to diagnose a metabolic cause of coma. Once diagnosis is established, the management becomes easier. On detailed investigation, she was found to have severe hypercalcemia with corrected serum calcium level being 18.2 mg%. Serum vitamin D level was also high, 150 ng/ml. On probing for detailed history, it was revealed that she had been taking a weekly dose of 60,000 IU of vitamin D for nearly 5 years. In recent times, association of various diseases has been reported with vitamin D deficiency. Vitamin D supplementation seems justified in this patient considering her age and the fact that she had rheumatoid arthritis; she was also taking corticosteroid. The recommended dose of vitamin D has been 400–800 IU/day. However, it is often prescribed at a dose of 60,000 IU/week, and sometimes patients self-medicate. There are no guidelines available for prescribing vitamin D at such a strength. Though safety of vitamin D has been established, vitamin D toxicity can occur sporadically with serious consequences. This was managed with intravenous fluids and diuretics. The patient also required subcutaneous calcitonin and low dose of corticosteroid, in addition to a session of haemodialysis for her hypercalcemia. The patient’s sensorium gradually improved as her calcium level returned to normalcy.

Zilebesiran, an RNA Interference Therapeutic Agent for Hypertension

BackgroundAngiotensinogen is the sole precursor of angiotensin peptides and has a key role in the pathogenesis of hypertension. Zilebesiran, an investigational RNA interference therapeutic agent with a prolonged duration of action, inhibits hepatic angiotensinogen synthesis.MethodsIn this phase 1 study, patients with hypertension were randomly assigned in a 2:1 ratio to receive either a single ascending subcutaneous dose of zilebesiran (10, 25, 50, 100, 200, 400, or 800 mg) or placebo and were followed for 24 weeks (Part A). Part B assessed the effect of the 800-mg dose of zilebesiran on blood pressure under low- or high-salt diet conditions, and Part E the effect of that dose when coadministered with irbesartan. End points included safety, pharmacokinetic and pharmacodynamic characteristics, and the change from baseline in systolic and diastolic blood pressure, as measured by 24-hour ambulatory blood-pressure monitoring.ResultsOf 107 patients enrolled, 5 had mild, transient injection-site reactions. There were no reports of hypotension, hyperkalemia, or worsening of renal function resulting in medical intervention. In Part A, patients receiving zilebesiran had decreases in serum angiotensinogen levels that were correlated with the administered dose (r=−0.56 at week 8; 95% confidence interval, −0.69 to −0.39). Single doses of zilebesiran (≥200 mg) were associated with decreases in systolic blood pressure (>10 mm Hg) and diastolic blood pressure (>5 mm Hg) by week 8; these changes were consistent throughout the diurnal cycle and were sustained at 24 weeks. Results from Parts B and E were consistent with attenuation of the effect on blood pressure by a high-salt diet and with an augmented effect through coadministration with irbesartan, respectively.ConclusionsDose-dependent decreases in serum angiotensinogen levels and 24-hour ambulatory blood pressure were sustained for up to 24 weeks after a single subcutaneous dose of zilebesiran of 200 mg or more; mild injection-site reactions were observed. (Funded by Alnylam Pharmaceuticals; ClinicalTrials.gov number, NCT03934307; EudraCT number, 2019-000129-39.)

Optimization of Drug-Linker to Enable Long-term Storage of Antibody-Drug Conjugate for Subcutaneous Dosing.

To facilitate subcutaneous dosing, biotherapeutics need to exhibit properties that enable high-concentration formulation and long-term stability in the formulation buffer. For antibody-drug conjugates (ADCs), the introduction of drug-linkers can lead to increased hydrophobicity and higher levels of aggregation, which are both detrimental to the properties required for subcutaneous dosing. Herein we show how the physicochemical properties of ADCs could be controlled through the drug-linker chemistry in combination with prodrug chemistry of the payload, and how optimization of these combinations could afford ADCs with significantly improved solution stability. Key to achieving this optimization is the use of an accelerated stress test performed in a minimal formulation buffer.

Evolution of Symptoms After Ustekinumab Induction Therapy in Patients With Crohn’s Disease

Ustekinumab is an effective treatment of Crohn's disease (CD). Of interest to patients is knowing how soon symptoms may improve. We analyzed ustekinumab response dynamics from the ustekinumab CD trials. Patients with CD received intravenous induction with ustekinumab ∼6 mg/kg (n= 458) or placebo (n= 457). Week 8 ustekinumab responders received subcutaneous ustekinumab 90 mg as the first maintenance dose or as an extended induction dose for nonresponders. Patient-reported symptom changes (stool frequency, abdominal pain, general well-being) within the first 14 days and clinical outcomes through week 44 were evaluated using the CD Activity Index. After ustekinumab infusion, stool frequency improvement was significantly (P < .05) greater than placebo on day 1 and for all patient-reported symptoms by day 10. In patients with no history of biologic failure or intolerance, cumulative clinical remission rates increased from 23.0% at week 3 to 55.5% at week 16 after the subcutaneous dose at week 8. Corresponding cumulative rates for patients with a history of biologic failure or intolerance increased from 12.9% to 24.1%. Neither change from baseline in CD Activity Index score nor week 8 ustekinumab pharmacokinetics were associated with week 16 response. Among all patients who received subcutaneous ustekinumab 90 mg q8w, up to 66.7% were in clinical response at week44. Ustekinumab induction provided symptom relief by day 1 post-infusion. Following ustekinumab infusion and a subcutaneous 90 mg injection, clinical outcomes continued to increase through week 16 and up to week 44. Regardless of week 8 clinical status or ustekinumab pharmacokinetics, patients should receive additional treatment at week8. gov numbers, NCT01369329, NCT01369342, and NCT01369355.

The effects of chronic high-dose morphine on microgliosis and the microglial transcriptome in rat spinal cord.

Background: Opioids are efficacious and safe analgesic drugs in short-term use for acute pain but chronic use can lead to tolerance and dependence. Opioid-induced microglial activation may contribute to the development of tolerance and this process may differ between males and females. A link is suggested between this microglial activation and inflammation, disturbances of circadian rhythms, and neurotoxic effects. We set out to further delineate the effects of chronic morphine on pain behaviour, microglial and neuronal staining, and the transcriptome of spinal microglia, to better understand the role of microglia in the consequences of long-term high-dose opioid administration. Experimental Approach: In two experiments, we administered increasing subcutaneous doses of morphine hydrochloride or saline to male and female rats. Thermal nociception was assessed with the tail flick and hot plate tests. In Experiment I, spinal cord (SC) samples were prepared for immunohistochemical staining for microglial and neuronal markers. In Experiment II, the transcriptome of microglia from the lumbar SC was analysed. Key Results: Female and male rats had similar antinociceptive responses to morphine and developed similar antinociceptive tolerance to thermal stimuli following chronic increasing high doses of s.c. morphine. The area of microglial IBA1-staining in SC decreased after 2weeks of morphine administration in both sexes. Following morphine treatment, the differentially expressed genes identified in the microglial transcriptome included ones related to the circadian rhythm, apoptosis, and immune system processes. Conclusions: Female and male rats showed similar pain behaviour following chronic high doses of morphine. This was associated with decreased staining of spinal microglia, suggesting either decreased activation or apoptosis. High-dose morphine administration also associated with several changes in gene expression in SC microglia, e.g., those related to the circadian rhythm (Per2, Per3, Dbp). These changes should be considered in the clinical consequences of long-term high-dose administration of opioids.

365 Phase 2 trial in progress—lirentelimab in adults with moderate-to-severe atopic dermatitis inadequately controlled by topical treatments

Abstract Atopic dermatitis (AD) is a chronic pruritic inflammatory dermatitis that affects approximately 16.5 million (7.3%) adults in the USA, of which around 6.6 million (40%) have moderate-to-severe disease. The current standard of care includes topical treatments supplemented with corticosteroids with and without immunosuppressants. Patients with AD inadequately controlled by topical treatments and immunosuppressants are often treated with biologics and/or JAK inhibitors. Some patients do not have an adequate clinical response or are unable to tolerate these treatments prompting the need for novel therapies. Mast cells (MCs) and eosinophils (eos) are implicated in the pathogenesis of AD. Sialic acid-binding Ig-like lectin (Siglec)-8 is expressed selectively on MCs and eos; engagement with an antibody results in broad inhibition of MC activation and eos depletion. Lirentelimab (AK002) is a humanized, nonfucosylated IgG1 monoclonal antibody that binds specifically to Siglec-8. Preclinical data demonstrate that lirentelimab can broadly inhibit multiple modes of MC stimulation that drive AD pathogenesis. Lirentelimab, administered every 4 weeks as an infusion (IV), has been tested in over 600 healthy volunteers and patients with inflammatory and allergic diseases; in those with concomitant AD, improvements in AD were observed. Overall, lirentelimab IV has been well-tolerated; the most common AE being infusion related reactions (IRRs) typically associated with the initial infusions. In a phase 1 study, a subcutaneous (SC) formulation of lirentelimab was well-tolerated with no IRRs. The results of these studies provide a strong rationale for conducting this phase 2 proof-of-concept, randomized, double-blind, placebo-controlled study of lirentelimab SC in adults with moderate-to-severe AD inadequately controlled by topical treatments (NCT05155085, ‘ATLAS’). Adults (18–80 years) are eligible for screening if they have had chronic AD presented for ≥3 years with moderate-to-severe symptoms defined as: Eczema Area and Severity Index (EASI) score ≥16, involvement of ≥10% of the Body Surface Area and Investigator Global Assessment (IGA) score ≥3, documented recent history of inadequate response to treatment with medications such as topical corticosteroids, calcineurin inhibitors, JAK inhibitors or PDE4 inhibitors (crisaborole), and biologic-naïve or biologic-exposed (secondary loss of response, intolerance or lack of access). The complete list of inclusion/exclusion criteria will be presented. One hundred and thirty patients will be randomized 1 : 1 to receive either seven SC injections of 300-mg lirentelimab or placebo every 2 weeks and then followed for 12 weeks after the last dose. The primary endpoint is the proportion of patients who achieve EASI-75 at week 14 (2 weeks after last dose) compared with baseline. All patients who receive study medication will be included in the safety analysis. Secondary endpoints include percent change in EASI at week 14 from baseline and proportion of patients with IGA of 0 or 1 and a 2-point improvement at week 14 compared with baseline. Patients who complete the study (Day 99 visit of double-blind period) will be given the option to enroll in an open-label extension for seven doses of 300-mg lirentelimab SC for 12 weeks. Qualified participants will receive medication, lab tests and medical care related to the study at no cost. This study is currently enrolling in the USA and Germany. Please visit clinicaltrials.gov (NCT05155085) or email atlas.info@allakos.com to learn more about this study.

75-LB: CT-388, a Novel Once-Weekly Dual GLP-1 and GIP Receptor Modulator, Is Safe, Well-Tolerated, and Produces More than 8% Weight Loss in Four Weeks in Overweight and Obese Adults

GLP-1 and GIP can deliver complementary pharmacology when combined. CT-388 is a biased dual GLP-1 and GIP receptor modulator that exhibits no to minimal β-arrestin coupling at either receptor, designed for once-weekly subcutaneous dosing. A Phase 1, randomized, placebo-controlled, double-blind study was conducted where single ascending doses (0.5-7.5 mg) and multiple ascending doses (MAD; doses 5-12 mg via titration in 3 cohorts) for 4 weeks were administered to overweight/obese adults. Primary objective was to investigate safety and tolerability of CT-388. Total of 64 participants (men/women: 28/36; median age/BMI: 34 years/33 kg/m2) received at least 1 dose of CT-388 or placebo. PK profile was investigated over a wide dose range (0.5-12 mg) and supports once-weekly administration. Percent change in body weight from baseline at Day 29 was dose responsive and significantly greater in CT-388 treated participants versus placebo (p &amp;lt;0.0001) across the 3 MAD cohorts with Least Square Mean difference [95% CI] of -4.8% [-6.3, -3.3], -6.4% [-7.8, -5.0], and -8.5% [-10.4, -6.7]. Mean percent decrease from baseline at Day 23 in fasting glucose (↓8-10%), insulin (↓20-26%), HOMA-IR (↓26-33%), AUC0-120min glucose (↓26-28%) and AUC0-120min insulin (↓33-58%) during oGTT were seen in cohorts dosed up to 12 mg CT-388, suggestive of improved insulin sensitivity. Over the 4-week treatment period, CT-388 was generally well tolerated with no treatment-related discontinuations. The most frequent side effects reported were gastrointestinal (decreased appetite, nausea, vomiting, diarrhea), which were mostly mild in severity. In summary, CT-388 delivers clinically meaningful weight loss and metabolic control with a favorable tolerability profile. These data warrant further clinical evaluation of CT-388, possibly with minimal to no titration, for the treatment of obesity, T2DM, and other weight-related comorbidities. Disclosure M. Chakravarthy: Employee; Carmot Therapeutics, Inc. F. A. Argüelles-Tello: Research Support; Carmot Therapeutics, Inc. A. A. Sun: None. M. Elliott: Employee; Carmot Therapeutics, Inc. L. Acosta: None. J. E. Rankin: None. S. K. Hansen: Board Member; Carmot Therapeutics, Inc.

Perspectives in weight control in diabetes – BI 456906

Glucagon-like peptide-1 receptor (GLP-1R) agonists are approved treatments for Type 2 diabetes mellitus, with liraglutide and semaglutide also approved for the treatment of obesity. The natural gut hormone oxyntomodulin is a weak dual agonist of the glucagon receptor (GCGR) and GLP-1R. Development of poly-agonists mimicking oxyntomodulin, such as the novel dual GCGR/GLP-1R agonist BI 456906, represents an important step towards a more effective treatment for people with Type 2 diabetes mellitus and obesity. BI 456906 is a 29-amino acid peptide derived from glucagon, with the incorporation of potent GLP-1 activities. It contains a C18 diacid which mediates binding to albumin, thereby prolonging the half-life to enable once-weekly subcutaneous dosing. The utilisation of GCGR agonism aims to enhance body weight-lowering effects by increasing energy expenditure in addition to the anorectic action of GLP-1R agonists. Glucose-lowering efficacy of BI 456906 has been demonstrated in a Phase II trial in patients with Type 2 diabetes mellitus and obesity and was associated with clinically meaningful body weight loss. These data highlight the potential of dual GCGR/GLP-1R agonism for reducing glycated haemoglobin and body weight in patients with Type 2 diabetes mellitus, and for greater therapeutic efficacy compared with GLP-1R agonism alone.

#4024 PHASE 1 STUDY OF THE SAFETY, TOLERABILITY, PHARMACOKINETICS, AND PHARMACODYNAMICS OF POVETACICEPT FOR AUTOIMMUNE GLOMERULONEPHRITIDES

Abstract Background and Aims B cell activating factor (BAFF) of the tumor necrosis factor (TNF) family and a proliferation-inducing ligand (APRIL), cytokines which bind and signal through BAFF-R, transmembrane activator and CAML interactor (TACI), and/or B cell maturation antigen (BCMA) on B cells, play overlapping and non-redundant roles in B cell development, proliferation, function, and survival. Therapeutic agents targeting BAFF and/or APRIL have demonstrated promising clinical potential in autoantibody-related glomerulonephritides (GN) such as lupus nephritis (LN), immunoglobulin (Ig) A nephropathy (IgAN), membranous nephropathy, and other B cell-related diseases such as systemic lupus erythematosus (SLE); however, there is still need for more safe and efficacious therapies. Povetacicept (ALPN-303) is an Fc fusion protein of an engineered TACI variant TNFRSF domain (vTD) with enhanced affinity for APRIL and BAFF which mediates more potent inhibitory activity than wild type (WT) TACI-Fc or BAFF- or APRIL-specific antibodies. In preclinical studies, povetacicept demonstrated enhanced pharmacokinetic (PK) and immunomodulatory properties vs. WT TACI-Fc, which may translate to lower and/or less frequent doses in humans. Povetacicept also suppressed autoantibodies, renal IgG deposition, and nephritis in mouse models. Povetacicept may therefore significantly improve clinical outcomes in autoantibody-mediated GNs and other B cell-related diseases. Method In this first-in-human study (NCT05034484), 66 healthy adult volunteers were randomized 4:2 into single ascending dose cohorts of intravenous (IV) or subcutaneous (SC) povetacicept or placebo. Participants were followed to assess safety and PK, circulating Ig, galactose-deficient IgA1 (Gd-IgA1), and circulating leukocyte populations. Results Povetacicept has been well tolerated in all cohorts evaluated as single IV or SC doses of up to 960 mg. Overall, it exhibits dose-related PK and expected pharmacodynamic (PD) effects, including dose-related reductions in serum IgA, IgM, IgG, and Gd-IgA1 (Figure 1), and in circulating antibody-secreting cells (ASC; plasmablasts and plasma cells) (Figure 2). In the same setting, these PD effects appear greater than those reported for WT TACI-Fc molecules and appear to be saturated at doses ≥80 mg. Coverage of free APRIL was maintained for 2-3 weeks with 80 mg and ≥4 weeks with 240 mg, respectively. The most frequent adverse event has been mild headache. To date, there have been no imbalances of infections between placebo and povetacicept groups, no treatment-related reactions other than mild injection site pain, and no adverse trends in safety laboratories. Conclusion To date, povetacicept has demonstrated acceptable safety and tolerability as single IV or SC doses, exhibiting dose-dependent PK and PD that appear to differentiate favorably vs WT TACI-Fc. Based on the magnitude and duration of the observed PD effects, dose regimens of 80-240 mg SC every 4 weeks are anticipated for use in future studies. Overall, the study findings support future clinical development of povetacicept in multiple autoantibody-related GN, as well as other B cell- and/or autoantibody-related diseases such as SLE and autoantibody-associated cytopenias.

#4033 RAPID COMPLEMENT INHIBITION WITH THE C5 INHIBITOR CROVALIMAB: A TIMING ANALYSIS USING ANIMAL MODEL AND COMPOSER TRIAL DATA

Abstract Background and Aims Atypical haemolytic uraemic syndrome (aHUS) is a life-threatening disease, characterised by acute kidney injury, thrombocytopenia, and microangiopathic haemolytic anaemia due to complement dysregulation. Currently approved treatments for aHUS include the anti-complement C5 monoclonal antibodies eculizumab and ravulizumab. Although currently approved therapeutic options are effective at both inhibiting complement-mediated thrombotic microangiopathy (TMA) and improving renal function, the regular intravenous (IV) infusion dosing regimens impose a significant treatment burden, particularly in paediatric patients (pts) with aHUS, who may need dialysis in the future due to chronic kidney injury and would benefit from preservation of vascular access. Crovalimab is a novel monoclonal antibody developed against complement C5, engineered to allow for small volume subcutaneous (SC) self-injection every 4 weeks, in a weight-based dosing regimen [1]. In the adaptive Phase I/II COMPOSER trial (NCT03157635) evaluating crovalimab in pts with paroxysmal nocturnal haemoglobinuria (PNH), that, like aHUS, is a complement disorder characterised by uncontrolled complement activation, crovalimab has shown maintained disease control and was well tolerated after a median exposure of 3 years. Further, a Phase III study evaluating crovalimab in previously untreated pts met its primary efficacy endpoints [2]. The efficacy and safety of crovalimab are currently being evaluated in adult and paediatric pts with aHUS, either treatment-naive or switching from another complement inhibitor, in the ongoing Phase III single-arm COMMUTE-a and COMMUTE-p trials [3]. Due to the rapidly progressing nature of aHUS, pts experiencing a TMA require rapid suppression of complement activation upon diagnosis to shut down the complement cascade and avoid further kidney deterioration and organ damage. Here, data from in vivo models and the COMPOSER trial were used to determine the time to complete complement inhibition after first crovalimab IV dose. Method Crovalimab's ability to suppress C5 function and complement activity rapidly was initially studied in in vivo models of cynomolgus monkeys. This study assessed the pharmacokinetics and pharmacodynamics of crovalimab in these monkeys after a single IV or SC dose. As part of this study, four animals per group were evaluated with single 4 mg/kg IV and 20 mg/kg IV doses. Crovalimab was also evaluated in the four-part, Phase I/II COMPOSER trial. Part 2 (n = 10) and Part 4A (n = 8) enrolled pts with PNH who were naive to complement inhibition. Pts in Part 2 received crovalimab 375 mg IV on Day 1, 500 mg IV on Day 8, 1000 mg IV on Day 22 and 170 mg SC weekly from Day 36 for 20 weeks. Pts in Part 4A received an optimised crovalimab dosing regimen of 1000 mg IV on Day 1, 340 mg SC on Days 2, 8, 15, and 22 and 680 mg SC Q4W from Day 29 onwards for 20 weeks. Crovalimab concentration, free C5 and complement activity were measured using validated assays in cynomolgus monkeys and in pts with PNH. Results Compared with baseline values in cynomolgus monkeys, a single crovalimab IV dose of 4 mg/kg reduced mean free C5 concentration by 99.6% and terminal complement activity by 81.4%, within 5 minutes after administration. In treatment-naive pts from COMPOSER Parts 2 and 4, mean free C5 concentration dropped to below 1 μg/mL, indicating a high level of target engagement within 1–6 hours from the first IV dose (Figure 1). Correspondingly, inhibition of terminal complement activity was reached within 1 hour, with values near or below the lower limit of quantification (10 U/mL; Figure 2). Complete complement blockade was generally maintained long-term, up to Week 20, in both Parts 2 and 4, regardless of dose. Conclusion Crovalimab induced a complete, rapid, and sustained blockade of terminal complement activity in both non-human primates and pts with PNH, within hours from first dose. The dosing schedule of crovalimab included an initial IV loading dose, allowing for a rapid onset of action, followed by a convenient long-term SC maintenance regimen.

Dosing Patterns of Dulaglutide and Semaglutide in Patients with Type 2 Diabetes in the United Kingdom and Germany: A Retrospective Cohort Study.

As new glucagon-like peptide 1 receptor agonist (GLP-1 RA) formulations are available, the aim of this study was to understand dulaglutide and subcutaneous (s.c.) semaglutide dosing patterns in people with type 2 diabetes mellitus (T2DM) in the UK and Germany as well as oral semaglutide in the UK. Adults with evidence of T2DM and a prescription of dulaglutide or semaglutide between August 2020 and December 2021 were identified using the IQVIA Longitudinal Prescription Data (LRx). Patients were divided into cohort 1 (incident users) and cohort 2 (prevalent users) based on previous exposure to GLP-1 RAs and were followed up to 12months post-index. During the patient selection window in Germany and the UK, 368,320 and 123,548 patients respectively received at least one prescription of a study GLP-1 RA. Among dulaglutide users in Germany at 12months post-index, the 1.5-mg dosage formulation was the most common for both cohort 1 (65.6%) and 2 (71.2%). Among s.c. semaglutide users at 12months post-index, 39.2% and 58.4% of cohort 1 received 0.5mg and 1.0mg, respectively. In the UK, at 12months post-index, the most common dulaglutide dosage formulation was 1.5mg (71.7% cohort 1 and 80.9% cohort 2). Among s.c. semaglutide users at 12months post-index, 0.5- and 1.0-mg formulations were the most common for both cohort 1 (38.9% and 56.0%, respectively) and cohort 2 (29.5% and 67.1%, respectively). Prescribing of the more recently introduced 3.0- and 4.5-mg formulations for dulaglutide and oral semaglutide was also reported in the study. Dosing patterns of GLP-1 RAs, although similar between the UK and Germany, were heterogeneous over time. Given that the higher dulaglutide doses and oral semaglutide were recently introduced to the market, additional real-world evidence studies which include clinical outcomes is required.

Phase I, Randomized, Placebo-Controlled, Dose-Escalation Study of GB223, a Fully-Humanized Monoclonal Antibody to RANKL, in Healthy Chinese Adults.

GB223 is a novel, fully-humanized monoclonal antibody against the receptor activator of nuclear factor-kappa B ligand (RANKL). In this phase I study, the safety, tolerability, pharmacokinetics, pharmacodynamics, and immunogenicity of GB223 were investigated. This was a randomized, double-blinded, placebo-controlled, single-dose escalation study conducted in 44 healthy Chinese adults. Participants were randomly assigned to receive a single subcutaneous injection dose of 7, 21, 63, 119, or 140 mg of GB223 (n=34) or placebo (n=10) and were followed up for 140-252 days. The results of noncompartmental analysis showed that GB223 was slowly absorbed after dosing, with a time to reach maximum concentration (Tmax) ranging from 5 to 11 days. Serum GB223 concentrations decreased slowly, with a long half-life ranging from 7.91 to 19.60 days. A two-compartment Michaelis-Menten model was found to best describe the pharmacokinetics of GB223, and the absorption rate of GB223 differed between males (0.0146 h-1) and females (0.0081 h-1). Serum C-terminal telopeptide of type I collagen decreased significantly postdose, and the inhibition lasted 42-168 days. No deaths or drug-related serious adverse events occurred. The most frequent adverse events were blood parathyroid hormone increased (94.1%), blood phosphorus decreased (67.6%) and blood calcium decreased (58.8%). In the GB223 group, 44.1% (15/34) of subjects were antidrug antibody positive after dosing. In this study, we demonstrated for the first time that a single subcutaneous injection of GB223, from 7 to 140 mg, is safe and well tolerated in healthy Chinese subjects. GB223 has a nonlinear pharmacokinetic profile, and sex was a potential covariate that may affect the absorption rate of GB223. NCT04178044 and ChiCTR1800020338.

Lefitolimod (TLR agonist) and ipilimumab in patients with advanced solid tumors: A phase I trial.

2564 Background: The TLR-9 agonist lefitolimod (MGN1703) significantly increases the Th1 response in pre-clinical models and has demonstrated antitumor activity in early phase clinical trials. This trial assessed the safety and preliminary efficacy of the rational combination of lefitolimod with ipilimumab in patients (pts) with advanced solid tumors. Methods: This was a single-center, open-label, investigator-initiated, dose-finding, and expansion Phase I trial (ClinicalTrials.gov no. NCT0266877). Key inclusion criteria were histologically confirmed advanced solid tumor, ECOG performance status ≤ 2; age ≥ 18 years. Patients (pts) were enrolled at escalating doses of lefitolimod (15-120 mg/kg), subcutaneously (SC) administered weekly and a fixed dose of ipilimumab 3 mg/kg in a 3-week cycle. For the expansion cohort, lefitolimod was administered intratumorally (IT) at 15 mg weekly. The primary endpoint was safety and tolerability according to CTCAE v4.0, dose-limiting toxicity (DLT) and maximum tolerated dose (MTD)/recommended Phase II dose (RP2D) for the lefitolimod in this combination. The secondary endpoint was efficacy, evaluated according to Immune-Related Response Criteria (irRC). Exploratory endpoints included changes in the intra-tumoral T-cell microenvironment in pre- and post- treatment tumor biopsies and the correlation of immune biomarkers with response. Results: From March 2017 to February 2020, 28 pts were enrolled. Median age was 56 years (range: 19-92). Median prior lines of therapy was 4 (range: 0-12). Pts 1-4 received lefitolimod 15 mg SC, pts 5-8 received lefitolimod 30 mg SC, pts 9-12 received lefitolimod 60 mg SC, pts 13-19 received lefitolimod 120 mg SC, and pts 20-28 received lefitolimod 15 mg IT. Eleven pts had at least one treatment-related AE (TRAE). The most common TRAE was skin rash (n=4, 14.2%), followed by fatigue (n=3, 10.7%) and pruritis (n=2, 7.1%). There were no grade 4 or 5 AEs, none of the pts required dose reduction, and there was no discontinuation of treatment due to AEs. DLT and MTD were not reached and the RP2D was established as lefitolimod SC 120 mg weekly with ipilimumab 3mg/Kg every 3 weeks. Of the 28 pts, 8 pts had stable disease as best response. 7 pts could not be evaluated by irRC due to clinical progression before restaging, non-measurable lesions and withdrawal of consent. Paired biopsy samples were analyzed by flow cytometry. Pro-inflammatory immune conditioning of the tumor microenvironment by the combination of the TLR9 agonist MGN1703 in combination with ipilimumab showed increases in intra-tumoral CD8 T cell frequency, memory CD8 phenotype (CD45RO+) and proliferation (Ki67+). Conclusions: The combination of high subcutaneous doses or intra-tumoral administration of lefitolimod in combination with ipilimumab is safe and well tolerated in patients with advanced cancers, with preliminary antitumor activity observed. Clinical trial information: NCT0266877 .

Subcutaneous amivantamab (ami) in patients (pts) with advanced solid malignancies: The PALOMA study—Updated safety and identification of the recommended phase 2 dose.

9126 Background: Ami, an EGFR-MET bispecific antibody, is approved for pts with advanced EGFR exon 20 insertion non-small cell lung cancer after progression on platinum-based chemotherapy. Intravenous (IV) delivery is associated with infusion-related reactions (IRRs) in 67% of pts, requiring splitting the first dose over 2 days (Park Ann Oncol 2021;32[suppl_5]:S981). PALOMA (NCT04606381) is an ongoing phase 1b dose escalation study of subcutaneous (SC) ami ± rHuPH20 (a hyaluronidase that aids SC agent absorption) in pts with advanced solid tumors who may benefit from EGFR- or MET-directed therapy. Preliminary results showed SC ami was well tolerated, improved time and ease of administration, and meaningfully reduced IRRs (Krebs Cancer Res 2022;82:12_Supplement, CT198). We present updated safety results and identification of the recommended phase 2 dose (RP2D) for ami SC Q2W administration. Methods: PALOMA enrolled pts with various advanced solid tumors. Objectives were to evaluate administration feasibility, safety, and PK of low and high concentration formulations of ami SC (50 mg/mL ami ± rHuPH20 [Part 1; Cohorts 1a/b] and 160 mg/mL ami ± rHuPH20 [Part 2; Cohorts 2a/b, Cohort 3a, Cohort 5a]). Cohorts 1a/b and 2a/b received 1050 mg (1400 mg, ≥80 kg), Cohort 3a received 1600 mg (2240 mg, ≥80 kg), and Cohort 5a received 2560 mg (3360 mg, ≥80 kg). Cohorts 1-3 were dosed weekly for the first 4 weeks and Q2W thereafter. Cohort 5a was dosed weekly for the first 3 weeks and Q3W thereafter. Results: As of Jan 3, 2023, 81 pts were enrolled (16 pts in Part 1, 65 pts in Part 2) and majority had NSCLC (71; 88%). Median age was 64 years, 44 (54%) pts were female, and most pts were White (44; 54%) or Asian (34; 42%). Across all doses, IRRs were reported by 13 (16%) pts; all of grade 1-2. The most frequent manifestations of IRRs were chills (7%), pyrexia (7%), and asymptomatic tachycardia (4%). Treatment-emergent AEs (TEAEs) of rash were reported by 59 (73%) pts, with no grade ≥3. In total, 3 (4%) pts discontinued ami SC due to toxicity (2 pneumonitis, 1 asthenia). Grade ≥3 related TEAEs were reported by 3 (4%) pts (hypoalbuminemia, lymphopenia, hypertension). Full ami SC dosing on day 1 was feasible (≤7 min), obviating the need for split dosing. PK analysis confirmed that ami SC 1600 mg (2240 mg, ≥80 kg) Q2W resulted in similar exposure to the approved IV dose (1050 mg [1400 mg, ≥80 kg] Q2W). Compared to IV, ami SC resulted in lower Cmax and equal or higher Ctrough and AUC0-336h at Cycles 2 and 4. Based on these data, ami SC 1600 mg (2240 mg, ≥80 kg) was selected as the RP2D for ami SC Q2W administration. Conclusions: Ami SC was well tolerated with meaningful reductions in administration time and TEAEs. Ami SC provided a quantitative and qualitative improvement in the symptoms of IRRs vs historical IV rates. The identified RP2D for ami SC on the Q2W schedule achieved similar exposure as the approved IV dose. Clinical trial information: NCT04606381 .

Effects of heterologous platelet-rich plasma on liver enzymes, oxidative stress biomarkers and pancreatic microarchitecture in diabetic mice

Diabetes Mellitus (DM) is a chronic metabolic disorder associated with inflammation and oxidative stress. Platelet-rich plasma (PRP) has an established role in alleviating DM. The current study aimed to evaluate the effect of heterologous PRP on liver functioning, oxidative stress, and microarchitecture of pancreatic islets in diabetic mice. Twenty-four (24) healthy male (4–5 weeks old) albino mice were selected and grouped as G1, G2, G3, and G4 (n ​= ​6). G1 was control group and remained untreated while G2 (PRP group) was given a subcutaneous dose of PRP (0.5 ​mL/kg body weight) twice a week for four weeks. G3 and G4 were first given a single dose of alloxan intraperitoneally (150 ​mg/kg) to induce diabetes. PRP treatment (0.5 ​mL/kg body weight) was given to G4 only for four weeks (twice a week). Completion of experimentation was followed by blood collection through the retro-orbital puncture and dissection for pancreas excision. Sera were isolated and liver functioning and oxidative stress were assessed. ALT (P ​= ​0.0022) and AST (P ​= ​0.0010) level were found higher in G3 compared to control (G1). Conversely, PRP treatment reduced it significantly in G4. MDA (P ​< ​0.0001) and GSH (P ​< ​0.0001) also showed a statistically significant difference while CAT (P ​= ​0.5288) showed no significant difference among all groups. Furthermore, histological analyses revealed the deformation in the microarchitecture of the pancreas in G3 as it displayed vacuolated cytoplasm, damaged islets, and collagen deposition. PRP treatment restored the islets as it was observed in G4. Results revealed that PRP restored transaminases and oxidative stress biomarkers in DM and ameliorate pancreatic microarchitecture.

Peripheral Administration of the Kv1.3-Blocking Peptide HsTX1[R14A] Improves Cognitive Performance in Senescence Accelerated SAMP8 Mice

Increased expression of the voltage-gated potassium channel Kv1.3 in activated microglia, and the subsequent release of pro-inflammatory mediators, are closely associated with the progression of Alzheimer’s disease (AD). Studies have shown that reducing neuroinflammation through the non-selective blockade of microglial Kv1.3 has the potential to improve cognitive function in mouse models of familial AD. We have previously demonstrated that a potent and highly-selective peptide blocker of Kv1.3, HsTX1[R14A], not only entered the brain parenchyma after peripheral administration in a lipopolysaccharide (LPS)-induced mouse model of inflammation, but also significantly reduced pro-inflammatory mediator release from activated microglia. In this study, we show that microglial expression of Kv1.3 is increased in senescence accelerated mice (SAMP8), an animal model of sporadic AD, and that subcutaneous dosing of HsTX1[R14A] (1 mg/kg) every other day for 8 weeks provided a robust improvement in cognitive deficits in SAMP8 mice. The effect of HsTX1[R14A] on the whole brain was assessed using transcriptomics, which revealed that the expression of genes associated with inflammation, neuron differentiation, synapse function, learning and memory were altered by HsTX1[R14A] treatment. Further study is required to investigate whether these changes are downstream effects of microglial Kv1.3 blockade or a result of alternative mechanisms, including any potential effect of Kv1.3 blockade on other brain cell types. Nonetheless, these results collectively demonstrate the cognitive benefits of Kv1.3 blockade with HsTX1[R14A] in a mouse model of sporadic AD, demonstrating its potential as a therapeutic candidate for this neurodegenerative disease.

In Vivo Antiviral Activity of Baloxavir against PA/I38T-Substituted Influenza A Viruses at Clinically Relevant Doses.

Although the prevalence of polymerase acidic (PA)/I38T strains of influenza virus with reduced susceptibility to baloxavir acid is low, there is a possibility of emergence under selective pressure. Furthermore, the virus may be transmitted between humans. We investigated the in vivo efficacy of baloxavir acid and oseltamivir phosphate against influenza A subtypes H1N1, H1N1pdm09, and H3N2, with PA/I38T substitution, at doses simulating human plasma concentrations. A pharmacokinetic/pharmacodynamic analysis was performed to strengthen the validity of the findings and the applicability in a clinical setting. Although the antiviral effect of baloxavir acid was attenuated in mice infected with PA/I38T-substituted viral strains compared with the wild type (WT), baloxavir acid significantly reduced virus titers at higher-but clinically relevant-doses. The virus titer reduction with baloxavir acid (30 mg/kg subcutaneous single dose) was comparable to that of oseltamivir phosphate (5 mg/kg orally twice daily) against H1N1 and H1N1pdm09 PA/I38T strains in mice, as well as the H3N2 PA/I38T strain in hamsters. Baloxavir acid demonstrated an antiviral effect against PA/I38T-substituted strains, at day 6, with no further viral rebound. In conclusion, baloxavir acid demonstrated dose-dependent antiviral effects comparable to that of oseltamivir phosphate, even though the degree of lung virus titer reduction was diminished in animal models infected with PA/I38T-substituted strains.

22 Influence of Cell-Mediated Immune Response Classification on Performance of Heat-Stressed Dairy Cattle

Abstract The objective was to evaluate the influence of cell-mediated immune response classification on performance of dairy cattle experiencing conditions of heat stress on pasture. On d 0, pregnant, lactating dairy cows (n = 27; 18 Holstein and 9 Jersey) received a subcutaneous sensitization dose of killed Candida albicans (CA; 0.5 mg) with Quil-A adjuvant (0.75 mg) in 2.5 mL of PBS. Tail skin fold thickness (SFT) was measured with a Harpenden caliper on d 14 immediately following an intradermal injection of killed CA (0.1 mg in 0.5 mL of PBS) in the right tail skin fold and a control injection (PBS) in the left tail skin fold. On d 15, change in SFT was measured and used to assign cell-mediated immune response classifications (CMIR) as High (HR; &amp;gt;0.5 SD above the mean; n = 9), Intermediate (IR; within 0.5 SD above or below the mean; n = 10), or Low (LR; &amp;gt;0.5 SD below the mean; n = 8). Cows from each CMIR were assigned to 1 of 3 environmental conditions (EC): No Abatement (no heat abatement provided; n = 9), Shade (portable shade structure; n = 9), or Sprinklers (polyvinyl chloride sprinkler system; n = 9) for a 38-d period (July 1, 2019 to August 8, 2019). Daily Milk Yield (DMY), Reticulorumen Temperature (RT), Heat Stress Score (HSS; 0 = respiration rate of &amp;lt; 80 breaths/min to 4 = moribund, labored breathing), and instances of Mouth Open (MO), Tongue Out (TO), and Drool were recorded daily. Respiration rate (RR; breaths/min) was recorded 3 times/day (morning, midday, and late afternoon) and averaged on Monday, Wednesday, and Friday weekly. Body Condition Score (BCS; 1 = emaciated to 5 = obese) was recorded weekly. Data were analyzed using MIXED procedures of SAS specific for repeated measures with CMIR, EC, Breed, Time and 2-way interactions as fixed effects and Pen as random. Low exhibited the lowest HSS and BCS, but the greatest MO and TO instances (P &amp;lt; 0.02). High exhibited the greatest instances of Drool (P &amp;lt; 0.02). Heat stress score, MO, and Drool were greatest in the No Abatement group (P &amp;lt; 0.01). Respiration rate was greatest in the No Abatement and Shade groups relative to Sprinklers (P &amp;lt; 0.01). Daily milk yield, RT, and Drool were increased whereas HSS and RR were decreased in Holstein relative to Jersey cows (P &amp;lt; 0.01). Interactions (P &amp;lt; 0.05) included: CMIR by EC for DMY, HSS, RR, MO, TO, and Drool; CMIR by Breed for RT, DMY, and BCS; EC by Breed for RT, DMY, HSS, and RR; and EC by Time for DMY, HSS, BCS, RR, MO, and TO. There was an effect of Time for all variables (P &amp;lt; 0.05). Preliminary results suggest an influence of cell-mediated immune response classification on performance of dairy cattle experiencing conditions of heat stress and warrant future studies with an increased number of animals.

Pharmacokinetics and safety of candidate tocilizumab biosimilar CT-P47 versus reference tocilizumab: a randomized, double-blind, single-dose phase I study

ABSTRACT Background CT-P47 is a candidate tocilizumab biosimilar. This study assessed the pharmacokinetic (PK) equivalence of CT-P47 and European Union-approved reference tocilizumab (EU-tocilizumab) in healthy Asian adults. Research design and methods This double-blind, multicenter, parallel-group trial randomized healthy adults (1:1) to receive a single (162 mg/0.9 mL) subcutaneous dose of CT-P47 or EU-tocilizumab. The primary endpoint (Part 2) was PK equivalence by area under the concentration – time curve (AUC) from time zero to last quantifiable concentration (AUC0–last), AUC from time zero to infinity (AUC0–inf), and maximum serum concentration (Cmax). PK equivalence was concluded if 90% confidence intervals (CIs) for the ratios of geometric least-squares means (gLSMs) were within the 80–125% equivalence margin. Additional PK endpoints, immunogenicity, and safety were evaluated. Results In Part 2, 289 participants were randomized (146 CT-P47; 143 EU-tocilizumab); 284 received study drug. AUC0–last, AUC0–inf, and Cmax were equivalent between CT-P47 and EU-tocilizumab: 90% CIs for the ratios of gLSMs were within the 80–125% equivalence margin. Secondary PK endpoints, immunogenicity, and safety were comparable between groups. Conclusions CT-P47 demonstrated PK equivalence with EU-tocilizumab and was well tolerated, following a single dose in healthy adults. Clinical trial registration www.clinicaltrials.gov identifier is NCT05188378.