A Randomized, Double-Blind, Placebo-Controlled, Ascending Dose Phase 1 Study of AK002, a Novel Siglec-8 Selective Monoclonal Antibody, in Healthy Subjects

Specific molecular markers for various normal and pathogenic cell states and cell types provide knowledge of basic biological systems and have a direct application in targeted therapy. We describe a proteomic method based on the combination of new and improved phage display antibody technologies and mass spectrometry that allows identification of cell type-specific protein markers. The most important features of the method are (i) reduction of experimental noise originating from background binding of phage particles and (ii) isolation of affinity binders after a single round of selection, which assures a high diversity of binders. The method demonstrates, for the first time, the ability to detect, identify, and analyze both secreted and membrane-associated extracellular proteins as well as a variety of different cellular structures including proteins and carbohydrates. The optimized phage display method was applied to analysis of human skin keratinocytes resulting in the isolation of a panel of antibodies. Fourteen of these antibodies were further characterized, half of which predominantly recognized keratinocytes in a screen of a range of different cell types. Three cognate keratinocyte antigens were subsequently identified by mass spectrometry as laminin-5, plectin, and fibronectin. The combination of phage display technology with mass spectrometry methods for protein identification is a general and promising approach for proteomic analysis of cell surface complexity.

N-glycosylation status of purified beta-haptoglobin from sera of 17 patients, and from sera of 14 healthy volunteer subjects, was compared by blotting with various lectins and antibodies. Patients in this study were diagnosed as having colon cancer through histological examination of each tumor tissue by biopsy. Blotting index of serum beta-haptoglobin with Aleuria aurantia lectin (AAL) was clearly higher for cancer patients than for healthy subjects. No such distinction was observed for blotting with three other lectins and two monoclonal antibodies. To determine tumor-associated reactivity of AAL binding as compared to inflammatory processes in colonic tissues, beta-haptoglobin separated from sera of 5 patients with Crohn's disease (CD), and 4 patients with ulcerative colitis (UC), was studied. All these cases, except one case of UC, showed AAL index lower than that in cancer cases, similarly to healthy subjects. The higher AAL binding of beta-haptoglobin in colon cancer patients than in healthy subjects appeared to be due to alpha-L-fucosyl residue, since it was eliminated by bovine kidney alpha-fucosidase treatment. N-linked glycans of serum haptoglobin from colon cancer patients vs. healthy subjects were released by N-glycanase, fluorescence-labeled, and subjected to normal-phase high performance liquid chromatography (NP-HPLC). Glycan structures were determined based on glucose unit (GU) values and their changes upon sequential treatment with various exoglycosidases. Glycosyl sequences and their branching status of glycans from 14 cases of serum beta-haptoglobin were characterized. The identified glycans were sialylated or nonsialylated, bi-antennary or tri-antennary structures, with or without terminal fucosylation.

Tildrakizumab, a high-affinity humanized IgG1k antibody that selectively binds interleukin (IL)-23 p19 subunit of cytokine IL-23 and neutralizes its function, is under investigation for treatment of moderate-to-severe chronic plaque psoriasis. The objective of this analysis was to assess the pharmacokinetics, bioavailability and safety/tolerability of single ascending doses of tildrakizumab after intravenous (IV) and subcutaneous (SC) dosing in healthy subjects. P05661 was a phase 1, single-dose, randomized, placebo-controlled study of tildrakizumab IV doses of 0.1, 0.5, 3 and 10mg/kg, or placebo. P05776 was a phase 1, single-dose, randomized, placebo-controlled study of tildrakizumab SC doses of 50 or 200mg, or placebo. After either single IV or SC dosing, tildrakizumab exhibited slow systemic clearance (CL), limited volume of distribution and a long t1/2 . Both the Cmax and the area under the curve (AUC) increased proportionally with doses from 0.1 to 10mg/kg, or 50-200mg. The bioavailability of SC dosing was ~80% (90% CI: 62-103%) for 50mg and ~73% (90% CI: 46-115%) for 200mg, respectively, versus 0.5 and 3mg/kg IV. Across both studies, six of 43 evaluable subjects were positive for post-dose antidrug antibodies; two of these were positive for neutralizing antibodies. Most adverse events (AEs) were mild; the most frequent AEs included upper respiratory tract infection and headache. Single doses of tildrakizumab 0.1, 0.5, 3 and 10mg/kg administered IV or single doses of 50 and 200 mg administered SC were safe and well tolerated in healthy adult subjects.

Background: Paroxysmal nocturnal hemoglobinuria (PNH) is a rare, acquired genetic disorder caused by clonal mutations in the PIGA gene. Deficiency of PIGA protein renders erythrocytes susceptible to complement-mediated destruction, causing intravascular and extravascular hemolysis, which can lead to anemia, fatigue, thromboembolism, major organ damage, and death. Despite the approval of one anti-C3 and two anti-C5 therapies for PNH, high treatment costs limit market access in many countries. CAN106 is a novel anti-C5 monoclonal antibody designed for optimal pH-dependent binding to C5 and enhanced binding to FcRN to increase intracellular recycling and prolong its half-life. Aims: This first-in-human Phase 1 study aimed to evaluate the single-dose safety and tolerability, pharmacokinetics, pharmacodynamics, and immunogenicity of CAN106. Methods: This was a randomized, double-blind, placebo-controlled, single-ascending-dose study. We enrolled 31 healthy adult subjects at a single site in Singapore who received CAN106 (mg/kg) or placebo at one of the following 6 doses (mg/kg) and ratios: (0.25, n=1:0; 0.75, n=1:0; 2, n=3:2; 4, n=6:2; 8, n=6:2 or 12, n=6:2). The primary endpoint was the incidence of adverse events overall and by intensity, seriousness, type, and relatedness to study drug. Secondary endpoints were the pharmacokinetic characterization of CAN106, the pharmacodynamic effects on free C5 (target engagement) and CH50 (serum hemolytic activity), and the incidence of anti-drug antibodies. The original study duration of 196 days was shortened to 112 days based upon the observed half-life of CAN106 and the return of CH50 to normal values. Results: The mean age of subjects was 34 years, and 97% were male. All 31 subjects completed the study. Seven CAN106 subjects experienced 20 treatment-emergent adverse events (TEAEs), and three placebo subjects experienced five TEAEs. Most TEAEs were mild and not related to CAN106, and all resolved without any sequelae. Three CAN106 subjects experienced seven drug-related TEAEs at the two highest doses. Most were mild, none was serious, and all resolved without any sequelae. These events included a moderate infusion-related reaction that resolved upon discontinuing treatment; mild dizziness; and mild elevations in ALT, AST, hemoglobin, hematocrit, and RBC. CAN106 exposure (Cmax and AUC) was dose-proportional, linear, and had low inter-subject variability (<20% CV). The terminal elimination half-life was approximately 32 days. CAN106 led to dose-dependent reductions in free C5 and CH50 within 24 hours. At the 8 and 12 mg/kg doses, all subjects showed >99% reduction in free C5 and >90% inhibition of CH50, with the latter sustained for 2 to 4 weeks. Two subjects in the 4 mg/kg cohort tested positive for anti-drug antibodies at single time points (pre-dose and Day 28). Summary/Conclusion: CAN106 was safe and well-tolerated at single doses up to 12 mg/kg in healthy adult subjects. The dose-exposure relationship was linear, and the 32-day half-life in healthy subjects was similar to that of ravulizumab, which is dosed every 8 weeks in patients, suggesting the potential for an extended dosing interval. CAN106 led to rapid, dose-dependent reductions in free C5 and CH50, and at the two highest doses, achieved complete and sustained complement blockade (>90% inhibition of CH50) for up to 4 weeks. These promising results support further clinical development of CAN106 in PNH and other complement-mediated diseases.

Raxibacumab is a fully humanized monoclonal antibody that blocks the interaction of Bacillus anthracis toxins, thereby protecting target cells from its effects. Raxibacumab is approved in the USA for the treatment of adults and children with inhalational anthrax in combination with antibiotics, and for prophylaxis of inhalational anthrax. The aim of this investigation was to characterise the population pharmacokinetics and assess the effect of baseline demographic covariates on the disposition of raxibacumab. The data used for this analysis were obtained from 3 clinical trials and include 2229 blood samples from 322 healthy subjects who were randomised to receive a 40 mg/kg intravenous dose of raxibacumab over a period of 2.25 hours. Population pharmacokinetic modelling was performed using a nonlinear mixed effects approach. Secondary parameters of interest were the area under the curve, maximum concentration and the time of serum raxibacumab concentrations greater than or equimolar to the highest serum protective antigen concentrations observed for at least 28 days in any monkey challenged with B. anthracis that died. Raxibacumab exposure in healthy subjects was described by a 2-compartment model. Interindividual variability was estimated for all model parameters, whilst residual variability was described by a proportional and additive error model. Weight was the only influential covariate with significant effect on disposition parameters. A dose of 40 mg/kg provided comparable exposure across the overall healthy subject population. Interindividual variability in raxibacumab vs. time profiles could partially be accounted for by differences in body weight.

BackgroundUV-4 (N-(9’-methoxynonyl)-1-deoxynojirimycin, also called MON-DNJ) is an iminosugar small-molecule oral drug candidate with in vitro antiviral activity against diverse viruses including dengue, influenza, and filoviruses and demonstrated in vivo efficacy against both dengue and influenza viruses. The antiviral mechanism of action of UV-4 is through inhibition of the host endoplasmic reticulum-resident α-glucosidase 1 and α-glucosidase 2 enzymes. This inhibition prevents proper glycan processing and folding of virus glycoproteins, thereby impacting virus assembly, secretion, and the fitness of nascent virions.Methodology/Principal findingsHere we report a first-in-human, single ascending dose Phase 1a study to evaluate the safety, tolerability, and pharmacokinetics of UV-4 hydrochloride (UV-4B) in healthy subjects (ClinicalTrials.gov Identifier NCT02061358). Sixty-four subjects received single oral doses of UV-4 as the hydrochloride salt equivalent to 3, 10, 30, 90, 180, 360, 720, or 1000 mg of UV-4 (6 subjects per cohort), or placebo (2 subjects per cohort). Single doses of UV-4 hydrochloride were well tolerated with no serious adverse events or dose-dependent increases in adverse events observed. Clinical laboratory results, vital signs, and physical examination data did not reveal any safety signals. Dose-limiting toxicity was not observed; the maximum tolerated dose of UV-4 hydrochloride in humans has not yet been determined (>1000 mg). UV-4 was rapidly absorbed and distributed after dosing with the oral solution formulation used in this study. Median time to reach maximum plasma concentration ranged from 0.5–1 hour and appeared to be independent of dose. Exposure increased approximately in proportion with dose over the 333-fold dose range. UV-4 was quantifiable in pooled urine over the entire collection interval for all doses.Conclusions/SignificanceUV-4 is a host-targeted broad-spectrum antiviral drug candidate. At doses in humans up to 1000 mg there were no serious adverse events reported and no subjects were withdrawn from the study due to treatment-emergent adverse events. These data suggest that therapeutically relevant drug levels of UV-4 can be safely administered to humans and support further clinical development of UV-4 hydrochloride or other candidate antivirals in the iminosugar class.Trial registrationClinicalTrials.gov NCT02061358https://clinicaltrials.gov/ct2/show/NCT02061358.

Inhibiting cMet has potential therapeutic value in oncology. The Janssen WAVE Early Development unit, a team focused on efficient proof-of-concept evaluations, sought to rapidly assess the safety, pharmacokinetics, and pharmacodynamics of a novel, highly selective small molecule cMet TK inhibitor with a favorable preclinical safety profile. We conducted a placebo-controlled, randomized, double blind single ascending dose and multiple dose trial in 84 healthy males at Quotient Clinical LTD (UK). The trial design was adaptive in several respects: to expedite determination of an optimal formulation and to maintain sufficient systemic exposure to sustain target inhibition. In the single ascending dose phase, doses ranged from 6-350 mg, with early transition from a solution to capsule formulation at 18 mg. Two multiple dose cohorts received 60 mg twice daily for 7 days (inter-dose intervals of 4.5 and 12 hours). Target engagement biomarkers were plasma hepatocyte growth factor and soluble cMet. Evaluation of renal safety was of particular interest. Clinical evaluation, including both single ascending and multiple dose phases, was completed in only 6 months. All doses were safe and well tolerated. Pharmacokinetic analysis revealed dose-proportional increases in exposure. Dose escalation was limited by concentrations of a major metabolite that approached the ceiling established in preclinical toxicology studies. Renal toxicity was not identified. Initial evaluation of this oncology drug in healthy subjects offered the advantages of rapid trial enrollment and completion, robust placebo comparison data, absence of concomitant illness or laboratory abnormalities, rapid dose escalation in a carefully-monitored, controlled setting, a thorough preliminary safety evaluation, and reduced costs. This approach, enabled by the acceptable preclinical safety profile of the compound, efficiently produced high quality clinical safety and pharmacokinetic data to support compound progression decisions. Citation Format: A Dawn Millington, Sandra R. Chaplan, Zuleima Aguilar, Dennis M. Fisher, Jo Collier, Marielena Mata, Geert Mannens, Nico Goyvaerts, Vijay Peddareddigari, Chris Takimoto. Rapid evaluation of a novel small molecule cMet tyrosine kinase inhibitor in healthy subjects. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr CT309. doi:10.1158/1538-7445.AM2015-CT309

The recent London tragedy [1] induces a huge emotion in the clinical pharmacology world. As decided by the Medicines and Healthcare products Regulatory Agency, the main point is to redefine the acceptable conditions for fully humanized agonist monoclonal antibodies in man. Such anxiety could also be reinforced by the fact that three deaths in healthy subjects have recently been reported in the USA [2–4]. Therefore, based on all the available data, it is necessary to reassess whether Phase I – early drug development in healthy subjects – is still safe: In the literature, only 15 deaths have been published during the last 30 years in Western countries, although probably 100 000 healthy subjects are dosed every year; moreover, only three deaths could not have been avoided if accurate common rules had been strictly adhered to. However, there is probably a higher risk in elderly subjects [5, 6]. Recently, a register has been initiated in France under the responsibility of the ‘Club Phase I’ (http://www.clubphase1.org/). This annual register is focused on serious adverse events (SAEs) and is limited to healthy subjects – young and elderly, men and women – and to Phase I studies. SAE refers to good clinical practice (GCP): death, life-threatening, requires hospitalization, disability, congenital anomaly and medically important event. It was implemented in 2004 and 2005; 15 386 healthy subjects have been included and dosed during the 2-year period (see Table 1). Two deaths occurred: one motorbike accident in a young man within a wash-out period and one lung cancer in an elderly woman undetected at screening in a study without drug administration (biomarker pilot study). No deaths were related to the tested drug. Sixty other SAEs were reported (not including the two above-mentioned deaths) (see Table 2). Only eight SAEs were really clinically serious, i.e. worrying with high risk, three out of eight only being related to the study drug: rash and fever, agranulocytosis and long-lasting atrial fibrillation. The others were not related: unstable angina, peritonitis, thrombosis (retina central vein), coma (car accident) and acute leukemia. Total SAE incidence is 0.4%, including worrying SAE 0.05%, of which related SAE 0.02%. In young subjects SAE incidence is 0.3%, but it is much higher in the elderly 2%, particularly in elderly women (3%). This register reporting more than 90% of the actual French activity in healthy subjects suggests clear conclusions: Phase I is safe, without related deaths and only 0.02% worrying and related SAEs; but a higher risk incidence is reported in elderly subjects, particularly in women. A recent Japanese register draws the same conclusions: from 1993 to 2004, 95 780 subjects, no deaths, low incidence of SAEs [47 (0.05%)], of which only 23 (0.02%) were related to drugs, all reversible without sequelae [7]. Thus, one can conclude: The risk of SAE in a Phase I study is low when common safety rules are applied: quality of drug supplies, use of a relevant method for the choice of doses, prudent progression from dose to dose using stopping criteria defined in protocols and approved by an ethics committee, resuscitation track and permanent presence of trained physicians on site . . . all, in fact, from GCP. The new data suggest reinforcing safety rules in two areas: Drug administration practice: define the number of subjects to administer at the same time, especially for first-in-class compounds, parenteral route, or possible immunogenicity of the drug. The same prudent approach is relevant in the case of possible low predictability of animal data: CNS, immune targets, fully humanized and recombinant compounds. Elderly subjects: special scrutiny and a more prudent approach are required. The authors thank all the French Clinical Research Organizations and Centres d’Investigations Cliniques that have provided data.

The purpose of this study is to evaluate the pharmacokinetics, immunogenicity, safety, and tolerability of guselkumab, a human monoclonal antibody with high affinity and specificity for binding to interleukin-23. In this first-in-human, phase 1, randomized study, a single intravenous (IV; 0.03-10mg/kg) or subcutaneous (SC; 10-300mg) dose of guselkumab was administered to 47 healthy subjects, and a single SC dose (placebo, 10, 30, 100, 300mg) was administered to 24 patients with moderate-to-severe psoriasis. Mean maximum observed serum concentration and area under the zero-to-infinity serum concentration-time curve of guselkumab increased in an approximately dose-proportional manner over the dose range of 0.03-10mg/kg following a single IV administration or 10-300mg following a single SC administration. Mean clearance and volume of distribution ranged from 3.62-6.03mL/day/kg and 99.38-123.22mL/kg, respectively. Mean half-life ranged from 12 to 19days in healthy subjects and patients with psoriasis. Among guselkumab-treated subjects/patients, 1/30 (3.3%) healthy subjects in the IV group, 0/6 healthy subjects in the SC group, and 1/20 (5.0%) patients with psoriasis tested positive for antibodies to guselkumab. No clinically significant adverse events were identified in this study. Guselkumab pharmacokinetic profiles were generally comparable between healthy subjects and patients with psoriasis. Guselkumab, administered as an IV infusion or SC injection, was well tolerated in healthy subjects and patients with psoriasis.

Homozygous or compound heterozygous mutation of the CD36 gene (CD36-/-) in humans results in severe defects of the uptake of long-chain fatty acids (LCFAs) in the heart. Because the effect of a single mutation of this gene (CD36+/-) on the LCFA uptake is not known, it was evaluated in 29 subjects with the CD36 wild-type gene (WT) (6 healthy subjects, 10 patients with heart disease), CD36+/- (4 healthy subjects, 5 patients) and CD36-/- (4 patients). The CD36 genotype was identified in the coding region of genomic DNA, and the expression of CD36 protein was examined by flow cytometry after staining with monoclonal anti-CD36 antibody. The LCFA uptake in the heart was assessed as the radioactivity accumulation ratio of heart to mediastinum after intravenous administration of iodine-123 15-(p-iodophenyl)-3-R, S-methylpentadecanoic acid (H/M ratio). The H/M ratios in WT, CD36+/- and CD36-/- were 2.28 +/- 0.10, 1.90 +/- 0.06 and 1.40 +/- 0.11, respectively (p < 0.0001, among groups). The H/M ratio between healthy subjects and patients with heart disease for WT and CD36+/- did not differ significantly (ie, those of WT and CD36+/- in healthy subjects and patients were 2.29 +/- 0.08 vs 2.27 +/- 0.12 and 1.90+/- 0.07 vs 1.89 +/- 0.05, respectively). Not only CD36-/- but also CD36+/- resulted in a significant reduction of the LCFA uptake in the heart independent of heart disease, suggesting genotype dependency and that CD36 might be a fundamental determinant of myocardial LCFA uptake.

Dupilumab is a fully human monoclonal antibody directed against the interleukin (IL)‐4 receptor α subunit (IL‐4Rα) of IL‐4 heterodimeric type I and type II receptors that mediate IL‐4/IL‐13 signaling through this pathway. Blockade of these receptors broadly suppresses type 2 inflammation associated with atopic/allergic diseases, including atopic dermatitis and asthma. Six phase 1 studies investigated the pharmacokinetics, pharmacodynamics, safety, and tolerability of dupilumab in healthy subjects. Two randomized, double‐blind, placebo‐controlled, sequential studies assessed safety and tolerability of single escalating dupilumab doses administered intravenously or subcutaneously (one included various racial groups, and one included exclusively Japanese subjects); 3 randomized, parallel‐group, single‐dose studies compared the pharmacokinetic profiles of different dupilumab products and formulations after single subcutaneous doses; and one study assessed dupilumab administered as fast versus slow subcutaneous injections. Dupilumab concentrations in serum were measured in all studies, and total immunoglobulin E (IgE) and thymus‐ and activation‐regulated chemokine (TARC) concentrations were measured in 2 studies as pharmacodynamic markers. Across the phase 1 studies, dupilumab exhibited target‐mediated pharmacokinetics consisting of parallel linear and nonlinear elimination, with the target‐mediated phase highly dominated by nonlinearity at lower drug concentrations. Systemic exposure and tolerability of dupilumab were consistent irrespective of differences in product, formulation, or racial background. Dupilumab reduced circulating concentrations of total IgE and TARC, indicating blockade of IL‐4Rα–mediated signaling. Dupilumab had a favorable safety profile across the wide range of doses administered. Together, these findings support the continued development and use of dupilumab in treatment of type 2 diseases.

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An overview and discussion of clinical studies with IC14, a chimeric monoclonal antibody directed against human CD14 is presented. These studies include phase 1 trials in: (i) healthy subjects; (ii) healthy subjects challenged with lipopolysaccharide (LPS); and (iii) patients with severe sepsis. The results from clinical studies of IC14 demonstrate this chimeric monoclonal antibody diminishes the systemic inflammatory response directed against LPS and may have potential as a treatment to prevent organ dysfunction in patients with severe sepsis. Furthermore, IC14 treatment was generally well-tolerated in both healthy subjects and severe sepsis patients and did not increase the incidence of secondary infections. Analysis of pharmacology data indicate that higher doses of IC14 are required to saturate membrane-bound CD14 receptors effectively in patients with severe sepsis than in healthy subjects. Finally, the results of non-clinical studies indicate that IC14 should only be administered with concurrent antimicrobial therapy in patients with infection.

Selection of monoclonal antibodies for immunoaffinity chromatography: model studies with antibodies against soy bean trypsin inhibitor

Transient transforming growth factor β1 modulation in monocytes and natural killer cells following plasma or plasma-platelet apheresis donation procedures.