High-concentration Formulation Research Articles

Relative bioavailability, immunogenicity, and safety of two adalimumab-adbm formulations in healthy volunteers: a double-blind, randomized, single-dose, parallel-arm Phase I trial (VOLTAIRE-HCLF)

ABSTRACT Objective VOLTAIRE-HCLF compared the relative bioavailability of citrate-free high-concentration and reference formulations of the biosimilar adalimumab-adbm (Cyltezo®), including pharmacokinetic (PK) profiles, immunogenicity, and safety profiles in healthy volunteers. Methods Healthy volunteers (N = 200) aged 18–55 years and with body mass index of 18.5–29.9 kg/m2 and no prior exposure to adalimumab were randomized in a 1:1 ratio to receive a single subcutaneous injection of either adalimumab-adbm 40 mg/0.4 mL (high-concentration formulation) or 40 mg/0.8 mL (reference formulation). Participants completed 13 follow-up visits over 57 days, followed by a safety follow-up period of up to 70 days. Results The main PK parameters were similar for the high-concentration and reference groups. For all primary endpoints, the geometric mean ratios and 90% confidence intervals of AUC0–1344, AUC0–∞, and Cmax for both groups were entirely within the standard 80–125% bioequivalence acceptance range at 101.88% (93.31–111.23%), 105.38% (95.06–116.81%), and 91.29% (84.38–98.76%), respectively. There were no differences in the proportion of anti-drug antibody-positive participants or in the distribution of anti-drug antibody titers between the two formulations at any time point after drug dosing. Participants who were given the high-concentration formulation of adalimumab-adbm experienced a lower incidence of adverse events and local reactions than those who were given the reference formulation. Conclusions Overall, the high-concentration and reference adalimumab-adbm formulations had highly similar PK and immunogenicity profiles and were safe and well tolerated. Clinical trial registration NCT05203289

Automated optimization of the solubility of a hyper-stable α-amylase.

Most successes in computational protein engineering to date have focused on enhancing one biophysical trait, while multi-trait optimization remains a challenge. Different biophysical properties are often conflicting, as mutations that improve one tend to worsen the others. In this study, we explored the potential of an automated computational design strategy, called CamSol Combination, to optimize solubility and stability of enzymes without affecting their activity. Specifically, we focus on Bacillus licheniformis α-amylase (BLA), a hyper-stable enzyme that finds diverse application in industry and biotechnology. We validate the computational predictions by producing 10 BLA variants, including the wild-type (WT) and three designed models harbouring between 6 and 8 mutations each. Our results show that all three models have substantially improved relative solubility over the WT, unaffected catalytic rate and retained hyper-stability, supporting the algorithm's capacity to optimize enzymes. High stability and solubility embody enzymes with superior resilience to chemical and physical stresses, enhance manufacturability and allow for high-concentration formulations characterized by extended shelf lives. This ability to readily optimize solubility and stability of enzymes will enable the rapid and reliable generation of highly robust and versatile reagents, poised to contribute to advancements in diverse scientific and industrial domains.

'Totality of Evidence' Approach in the Development of GP2017, an Approved Adalimumab Biosimilar.

Hyrimoz®, (GP2017 [SDZ-ADL]), is a biosimilar to Humira® (REF-ADL). SDZ-ADL was approved in 2018 by both the United States Food and Drug Administration (US FDA) and European Medicines Agency (EMA) for the indications of REF-ADL not protected by orphan exclusivity. In 2023, the US FDA and EMA also approved a citrate-free high-concentration formulation (HCF) of SDZ-ADL. Approval of SDZ-ADL was based on data gathered using the US FDA, EMA and World Health Organization (WHO)-recommended step-wise Totality of Evidence approach. This approach is a robust dataset confirming high confidence in analytical, functional, pharmacokinetic (PK) and clinical biosimilarity between the biosimilar and reference medicine determined through analytical and clinical investigation. Evidence supporting the biosimilarity of SDZ-ADL and REF-ADL was reported at each stage of investigation. Comprehensive comparative analytical and functional assessments demonstrated that SDZ-ADL was analytically indistinguishable from REF-ADL in required critical quality attributes, including receptor binding. Phase I clinical data showed PK similarity of SDZ-ADL and REF-ADL in healthy volunteers, with similar safety, tolerability and immunogenicity profiles. Phase III confirmatory efficacy and safety studies, ADACCESS (included in US/EU dossiers) and ADMYRA (separate to US/EU dossiers), both confirmed that SDZ-ADL's efficacy, safety, and immunogenicity matched REF-ADL in all patient groups with no clinically meaningful differences. More recently, this data package was the basis for a citrate-free HCF of SDZ-ADL to be developed, and its PK, safety and immunogenicity were confirmed against the initially approved formulation of SDZ-ADL. Overall, the Totality of Evidence provided for biosimilar adalimumab, SDZ-ADL, confirmed the analytical, functional and clinical similarity of SDZ-ADL to REF-ADL, supporting its regulatory approval and providing a data bridge with which to evaluate and support the approval of citrate-free HCF SDZ-ADL for clinical use.

Correlating Surface Activity with Interface-Induced Aggregation in a High-Concentration mAb Solution.

Interface-induced aggregation resulting in protein particle formation is an issue during the manufacturing and storage of protein-based therapeutics. High-concentration formulations of therapeutic proteins are even more prone to protein particle formation due to increased protein-protein interactions. However, the dependence of interface-induced protein particle formation on bulk protein concentration is not understood. Furthermore, the formation of protein particles is often mitigated by the addition of polysorbate-based surfactants. However, the details of surfactant-protein interactions that prevent protein particle formation at high concentrations remain unclear. In this work, a tensiometer technique was used to evaluate the surface pressure of an industrially relevant mAb at different bulk concentrations, and in the absence and presence of a polysorbate-based surfactant, polysorbate 20 (PS20). The adsorption kinetics was correlated with subvisible protein particle formation at the air-water interface and in the bulk protein solution using a microflow imaging technique. Our results showed that, in the absence of any surfactant, the number of subvisible particles in the bulk protein solutions increased linearly with mAb concentration, while the number of protein particles measured at the interface showed a logarithmic dependence on bulk protein concentration. In the presence of surfactants above the critical micelle concentration (CMC), our results for low-concentration mAb solutions (10 mg/mL) showed an interface that is surfactant-dominated, and particle characterization results showed that the addition of the surfactant led to reduced particle formation. In contrast, for the highest concentration (170 mg/mL), coadsorption of proteins and surfactants was observed at the air-water interface, even for surfactant formulations above CMC and the surfactant did not mitigate subvisible particle formation. Our results taken together provide evidence that the ratio between the surfactant and mAb molecules is an important consideration when formulating high-concentration mAb therapeutics to prevent unwanted aggregation.

ProtT5 and random forests-based viscosity prediction method for therapeutic mAbs

Viscosity is a key characteristic of therapeutic antibodies for subcutaneous administration which requires low volume and high concentration formulations. It would be highly beneficial to accurately predict the viscosity of newly developed therapeutic antibodies in the early stages of development. In this work, a ProtT5-XL-UniRef50 (ProtT5) and Random Forests (RF)-based prediction method was proposed for accurately predicting the viscosity of monoclonal antibodies, with only corresponding sequences needed. Starting from the given heavy and light chain V-region sequences, corresponding features were first extracted from the ProtT5 pretrained model. Kernel principal analysis (Kernel-PCA) was then used for reducing the extracted 2048-D (1024-D for each sequence) feature vector to a reasonable level for efficient training of the RF-regressor. Then, the RF model was constructed on 40 commercially available therapeutic antibodies and tested with 3-folds cross-validation. Test results show that the model could reproduce the viscosity value at a high level (Pearson correlation coefficient (PCC) = 0.928). Performance on classifying high (>30 cP) and low (<30 cP) viscosity is much more satisfactory, the Accuracy (ACC) and the area under precision-recall curve (AUC) of the classification model from validation tests are 0.975 and 1.000, respectively. Compared to 5 existing state-of-the-art viscosity prediction methods, the proposed method performs best which would facilitate high concentration antibody viscosity high-throughput screening.

A plain language summary describing how two different concentrations of GP2017, a biosimilar adalimumab medicine, are associated with similar drug levels within the body.

This plain language summary explains, in simple terms, the results of a study from 2022 discussing a biosimilar medicine called GP2017 (called SDZ-ADL in this summary, sold as Hyrimoz®). This medicine is used to treat people with inflammatory conditions. This study investigated a new, high-concentration formulation of GP2017 (SDZ-ADL-HCF) in order to show that the high concentration option acts the same way in the body as SDZ-ADL. SDZ-ADL-HCF has been submitted for regulatory approval to health authorities on the basis of this study and was recently approved by the US Food and Drug Administration (FDA) and the European Medicines Agency (EMA) for people with the inflammatory conditions that SDZ-ADL is used to treat. This newly developed formulation provides the option for receiving injections less often with reduced volumes which can have a positive impact on the injection experience and increase patient convenience. This study looked at the pharmacokinetics of SDZ-ADL and SDZ-ADL-HCF, meaning it compared how the active medicine behaved in the body at different times after the injection of each of the formulations. The study also looked at how each formulation was recognized by the body's immune system (known as immunogenicity), and the side effects associated with each formulation. This study was randomly assigned and double-blinded, meaning that neither the participants nor the researchers knew which formulation each participant received. This reduces the risk of bias in the results. The study found that an injection of SDZ-ADL-HCF resulted in similar amounts of the medicine being present within the blood as an injection of SDZ-ADL. This information was needed for the approval of SDZ-ADL-HCF. Participants also experienced similar immune reactions and the number of participants with side effects was similar between both concentrations of medicine. The results confirmed that SDZ-ADL-HCF behaves in the same way in the body and is expected to have the same treatment effects as SDZ-ADL, while at the same time offering an improved formulation with a more positive injection experience and increased patient convenience.

Reversible protein complexes as a promising avenue for the development of high concentration formulations of biologics

High concentration formulations have become an important pre-requisite in the development of biological drugs, particularly in the case of subcutaneous administration where limited injection volume negatively affects the administered dose. In this study, we propose to develop high concentration formulations of biologics using a reversible protein-polyelectrolyte complex (RPC) approach. First, the versatility of RPC was assessed using different complexing agents and formats of therapeutic proteins, to define the optimal conditions for complexation and dissociation of the complex. The stability of the protein was investigated before and after complexation, as well as upon a 4-week storage period at various temperatures. Subsequently, two approaches were selected to develop high concentration RPC formulations: first, using up-concentrated RPC suspensions in aqueous buffers, and second, by generating spray-dried RPC and further resuspension in non-aqueous solvents. Results showed that the RPC concept is applicable to a wide range of therapeutic protein formats and the complexation-dissociation process did not affect the stability of the proteins. High concentration formulations up to 200 mg/mL could be achieved by up-concentrating RPC suspensions in aqueous buffers and RPC suspensions in non-aqueous solvents were concentrated up to 250 mg/mL. Although optimization is needed, our data suggests that RPC may be a promising avenue to achieve high concentration formulations of biologics for subcutaneous administration.

Predicting Colloidal Stability of High-Concentration Monoclonal Antibody Formulations in Common Pharmaceutical Buffers Using Improved Polyethylene Glycol Induced Protein Precipitation Assay.

Colloidal stability is an important consideration when developing high concentration mAb formulations. PEG-induced protein precipitation is a commonly used assay to assess the colloidal stability of protein solutions. However, the practical usefulness and the current theoretical model for this assay have yet to be verified over a large formulation space across multiple mAbs and mAb-based modalities. In the present study, we used PEG-induced protein precipitation assays to evaluate colloidal stability of 3 mAbs in 24 common formulation buffers at 20 and 5 °C. These prediction assays were conducted at low protein concentration (1 mg/mL). We also directly characterized high concentration (100 mg/mL) formulations for cold-induced phase separation, turbidity, and concentratibility by ultrafiltration. This systematic study allowed analysis of the correlation between the results of low concentration assays and the high concentration attributes. The key findings of this study include the following: (1) verification of the usefulness of three different parameters (Cmid, μB, and Tcloud) from PEG-induced protein precipitation assays for ranking colloidal stability of high concentration mAb formulations; (2) a new method to implement PEG-induced protein precipitation assay suitable for high throughput screening with low sample consumption; (3) improvement in the theoretical model for calculating robust thermodynamic parameters of colloidal stability (μB and εB) that are independent of specific experimental settings; (4) systematic evaluation of the effects of pH and buffer salts on colloidal stability of mAbs in common formulation buffers. These findings provide improved theoretical and practical tools for assessing the colloidal stability of mAbs and mAb-based modalities during formulation development.

Advancing Subcutaneous Dosing Regimens for Biotherapeutics: Clinical Strategies for Expedited Market Access.

In recent years, subcutaneous administration of biotherapeutics has made significant progress. The self-administration market for rheumatoid arthritis has witnessed the introduction of additional follow-on biologics, while the first subcutaneous dosing options for monoclonal antibodies have become available for multiple sclerosis. Oncology has also seen advancements with the authorization of high-volume subcutaneous formulations, facilitated by the development of high-concentration formulations and innovative delivery systems. Regulatory and Health Technology Assessment bodies increasingly consider preference data in filing dossiers, particularly in evaluating novel drug delivery methods. The adoption of a pharmacokinetic-based clinical bridging approach has become standard for transitioning from intravenous to subcutaneous administration. Non-inferiority studies with pharmacokinetics as the only primary endpoint have started deviating from traditional randomization schemes, favoring the subcutaneous route and comparing with historical intravenous data. While nonclinical and computational models made progress in predicting safety and immunogenicity for subcutaneously dosed antibodies, clinical trial evidence remains essential due to inter-individual variations and the impact of formulation parameters on anti-drug antibody formation. Ongoing technological advancements and the expanding knowledge base on pharmacokinetic-pharmacodynamic correlation across specialty areas are expected to further accelerate clinical development of subcutaneous biologics.

Effects of arginine in therapeutic protein formulations: a decade review and perspectives

Abstract Arginine (Arg) is a natural amino acid with an acceptable safety profile and a unique chemical structure. Arg and its salts are highly effective in enhancing protein refolding and solubilization, suppressing protein–protein interaction and aggregation and reducing viscosity of high concentration protein formulations. Arg and its salts have been used in research and 20 approved protein injectables. This review summarizes the effects of Arg as an excipient in therapeutic protein formulations with the focus on its physicochemical properties, safety, applications in approved protein products, beneficial and detrimental effects in liquid and lyophilized protein formulations when combined with different counterions and mechanism on protein stabilization and destabilization. The decade literature review indicates that the benefits of Arg overweigh its risks when it is used appropriately. It is recommended to add Arg along with glutamate as a counterion to high concentration protein formulations on top of sugars or polyols to counterbalance the negative effects of Arg hydrochloride. The use of Arg as a viscosity reducer and protein stabilizer in high concentration formulations will be the inevitable future trend of the biopharmaceutical industry for subcutaneous administration.

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.

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 .

Crystalline Antibody-Laden Alginate Particles: A Platform for Enabling High Concentration Subcutaneous Delivery of Antibodies.

Subcutaneous (SC) administration is a desired route for monoclonal antibodies (mAbs). However, formulating mAbs for small injection volumes at high concentrations with suitable stability and injectability is a significant challenge. Here, this work presents a platform technology that combines the stability of crystalline antibodies with injectability and tunability of soft hydrogel particles. Composite alginate hydrogel particles are generated via a gentle centrifugal encapsulation process which avoids use of chemical reactions or an external organic phase. Crystalline suspension of anti-programmed cell death protein1 (PD-1) antibody (pembrolizumab) is utilized as a model therapeutic antibody. Crystalline forms of the mAb encapsuled in the hydrogel particles lead to stable, high concentration, and injectable formulations. Formulation concentrations as high as 315mg mL-1 antibody are achieved with encapsulation efficiencies in the range of 89-97%, with no perceivable increase in the number of antibody aggregates. Bioanalytical studies confirm superior maintained quality of the antibody in comparison with formulation approaches involving organic phases and chemical reactions. This work illustrates tuning the alginate particles' disintegration by using partially oxide alginates. Crystalline mAb-laden particles are evaluated for their biocompatibility using cell-based in vitro assays. Furthermore, the pharmacokinetics (PK) of the subcutaneously delivered human anti-PD-1 mAb in crystalline antibody-laden alginate hydrogel particles in Wistar rats is evaluated.

AF4 and PEG Precipitation as Predictive Assays for Antibody Self-Association.

Monoclonal antibodies (mAbs) are often formulated as high-protein-concentration solutions, which in some cases can exhibit physical stability issues such as high viscosity and opalescence. To ensure that mAb-based drugs can meet their manufacturing, stability, and delivery requirements, it is advantageous to screen for and select mAbs during discovery that are not prone to such behaviors. It has been recently shown that both these macroscopic properties can be predicted to a certain extent from the diffusion interaction parameter (kD), which is a measure of self-association under dilute conditions.1 However, kD can be challenging to measure at the early stage of discovery, where a relatively large amount of a high-purity material, which is required by traditional methods, is often not available. In this study, we demonstrate asymmetric field-flow fractionation (AF4) as a tool to measure self-association and therefore identify antibodies with problematic issues at high concentrations. The principle lies on the ability to concentrate the sample close to the membrane during the injection mode, which can reach formulation-relevant concentrations (>100 mg/mL).2 By analyzing a well-characterized library of commercial antibodies, we show that the measured retention time of the antibodies allows us to pinpoint molecules that exhibit issues at high concentrations. Remarkably, our AF4 assay requires very little (30 μg) sample under dilute conditions and does not need extensive sample purification. Furthermore, we show that a polyethylene glycol (PEG) precipitation assay provides results consistent with AF4 and moreover can further differentiate molecules with issues of opalescence or high viscosity. Overall, our results delineate a two-step strategy for the identification of problematic variants at high concentrations, with AF4 for early developability screening, followed by a PEG assay to validate the problematic molecules and further discriminate between opalescence or high-viscosity issues. This two-step antibody selection strategy enables us to select antibodies early in the discovery process, which are compatible with high-concentration formulations.

An Intercompany Perspective on Practical Experiences of Predicting, Optimizing and Analyzing High Concentration Biologic Therapeutic Formulations

Developing high-dose biologic drugs for subcutaneous injection often requires high-concentration formulations and optimizing viscosity, solubility, and stability while overcoming analytical, manufacturing, and administration challenges. To understand industry approaches for developing high-concentration formulations, the Formulation Workstream of the BioPhorum Development Group, an industry-wide consortium, conducted an inter-company collaborative exercise which included several surveys. This collaboration provided an industry perspective, experience, and insight into the practicalities for developing high-concentration biologics. To understand solubility and viscosity, companies desire predictive tools, but experience indicates that these are not reliable and experimental strategies are best. Similarly, most companies prefer accelerated and stress stability studies to in-silico or biophysical-based prediction methods to assess aggregation. In addition, optimization of primary container-closure and devices are pursued to mitigate challenges associated with high viscosity of the formulation. Formulation strategies including excipient selection and application of studies at low concentration to high-concentration formulations are reported. Finally, analytical approaches to high concentration formulations are presented. The survey suggests that although prediction of viscosity, solubility, and long-term stability is desirable, the outcome can be inconsistent and molecule dependent. Significant experimental studies are required to confirm robust product definition as modeling at low protein concentrations will not necessarily extrapolate to high concentration formulations.

Stable High-Concentration Monoclonal Antibody Formulations Enabled by an Amphiphilic Copolymer Excipient.

Monoclonal antibodies are a staple in modern pharmacotherapy. Unfortunately, these biopharmaceuticals are limited by their tendency to aggregate in formulation, resulting in poor stability and often requiring low concentration drug formulations. Moreover, existing excipients designed to stabilize these formulations are often limited by their toxicity and tendency to form particles such as micelles. Here, we demonstrate the ability of a simple "drop-in", amphiphilic copolymer excipient to enhance the stability of high concentration formulations of clinically-relevant monoclonal antibodies without altering their pharmacokinetics or injectability. Through interfacial rheology and surface tension measurements, we demonstrate that the copolymer excipient competitively adsorbs to formulation interfaces. Further, through determination of monomeric composition and retained bioactivity through stressed aging, we show that this excipient confers a significant stability benefit to high concentration antibody formulations. Finally, we demonstrate that the excipient behaves as an inactive ingredient, having no significant impact on the pharmacokinetic profile of a clinically relevant antibody in mice. This amphiphilic copolymer excipient demonstrates promise as a simple formulation additive to create stable, high concentration antibody formulations, thereby enabling improved treatment options such as a route-of-administration switch from low concentration intravenous (IV) to high concentration subcutaneous (SC) delivery while reducing dependence on the cold chain.

S1057 A Randomized Phase I Study Comparing Pharmacokinetics, Safety and Immunogenicity of High Concentration Formulation (100 mg/mL) With GP2017 Formulation (50 mg/mL), an Adalimumab Biosimilar, in Healthy Male Subjects

Introduction: GP2017 (adalimumab-adaz) is an approved biosimilar of adalimumab (ADL)1, with a concentration of 50 mg/mL. A high-concentration formulation (HCF) (100 mg/mL) can reduce the injection volume, and consequently decrease the number of injections required for patients who need to administer a higher dose. A phase I study (randomized, double-blind, parallel, 2-arm study with healthy male subjects) was conducted to demonstrate pharmacokinetics (PK) comparability between a newly developed GP2017 citrate-free -HCF and the currently approved GP2017. Methods: Healthy male subjects were randomized to receive a single 40 mg s.c. injection of either GP2017 or GP2017-HCF. PK, safety, and immunogenicity were assessed over 72 days post-injection. Primary endpoints were maximum serum concentration (Cmax), area under the concentration–time curve from time zero to infinity (AUC0–inf) and area under the concentration–time curve from time zero to 360 hours (AUC0-360); area under the concentration–time curve from time zero to the last quantifiable concentration (AUC0–last) was assessed as a secondary endpoint. PK comparability was concluded when the 90% confidence intervals (CIs) of the ratios of geometric means were within the predefined comparability margin of 0.80 to 1.25. Results: 331 subjects were randomized, of which 330 received study treatment (162 GP2017-HCF; 168 GP2017). The 90% CI of the ratios (GP2017-HCF / GP2017) of geometric means for the PK endpoints (Cmax, AUCinf, AUC0-360 and AUC0–last) were all contained within the pre-defined comparability limits of 0.80 to 1.25 (Figure). Safety was similar across groups (Table). The numbers and proportions of subjects with positive anti-drug antibodies (ADA) and with neutralizing antibodies (NAb) were comparable overall (up to day 72: GP2017-HCF 69.1% / 63%; GP2017 64.9% / 61.3%, respectively) and at all individual visits. Conclusion: PK comparability between GP2017-HCF and GP2017 was demonstrated, supporting the development of the new formulation of GP2017. Safety and immunogenicity profiles were comparable between treatment groups and consistent with previously reported adalimumab data.Figure 1.: Forest plot to compare primary and secondary PK endpoints between the treatment groups Table 1. - TEAEs by primary system organ class (at least 2% of subjects in any treatment group) Primary system organ classPreferred term GP2017-HCFN=162n (%) GP2017N=168n (%) Number of subjects with at least 1 event 80 (49.4) 95 (56.5) General disorders and administration site conditions 55 (34.0) 69 (41.1) Investigations 16 (9.9) 24 (14.3) Musculoskeletal and connective tissue disorders 8 (4.9) 14 (8.3) Nervous system disorders 8 (4.9) 6 (3.6) Skin and subcutaneous tissue disorders 10 (6.2) 4 (2.4) Gastrointestinal disorders 8 (4.9) 3 (1.8) Respiratory, thoracic and mediastinal disorders 2 (1.2) 6 (3.6) Eye disorders 1 (0.6) 4 (2.4) Injury, poisoning and procedural complications 4 (2.5) 1 (0.6)

Antibody apparent solubility prediction from sequence by transfer learning

SummaryDeveloping therapeutic monoclonal antibodies (mAbs) for the subcutaneous administration requires identifying mAbs with superior solubility that are amenable for high-concentration formulation. However, experimental screening is often material and labor intensive. Here, we present a strategy (named solPredict) that employs the embeddings from pretrained protein language modeling to predict the apparent solubility of mAbs in histidine (pH 6.0) buffer. A dataset of 220 diverse, in-house mAbs were used for model training and hyperparameter tuning through 5-fold cross validation. solPredict achieves high correlation with experimental solubility on an independent test set of 40 mAbs. Importantly, solPredict performs well for both IgG1 and IgG4 subclasses despite the distinct solubility behaviors. This approach eliminates the need of 3D structure modeling of mAbs, descriptor computation, and expert-crafted input features. The minimal computational expense of solPredict enables rapid, large-scale, and high-throughput screening of mAbs using sequence information alone during early antibody discovery.

Pharmacokinetics, blood and urine profile effects, and injection site histopathology following three daily injections of subcutaneous high concentration buprenorphine in New Zealand white rabbits (Oryctolagus cuniculus)

Buprenorphine is a common analgesic administered to rabbits and high concentration formulations can reduce handling stress without sacrificing pain relief. The objective of this study was to evaluate the pharmacokinetics of high concentration buprenorphine and its metabolites following multiple subcutaneous doses in the rabbit. Laboratory variables (complete blood cell count , biochemistry profile, urinalysis) were compared for drug effects and injection sites were evaluated via histopathology. High concentration buprenorphine (HCB) was administered subcutaneously (0.24 mg/kg) to six, 17-week-old, intact female, New Zealand white rabbits ( Oryctolagus cuniculus ) for three doses, 24 hours apart. Two control animals received an equal volume of saline. Blood samples were collected at -72, 0, 0.33, 0.66, 1.5, 3, 6, 12, 24, 48, 72, 96, 120, and 144 hours. Buprenorphine and its metabolites were measured via liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS). Blood and urine profiles were collected prior to and following HCB administration and compared within and between groups. Injection sites and major organ systems were evaluated grossly and microscopically for drug effects. High concentration buprenorphine administered once every 24 hours for three doses had a variable accumulation index of 1.68 (1.24–2.29), reflecting variability in terminal half-life. Subsequently, the metabolites buprenorphine-3-glucuronide and norbuprenorphine-3-glucuronide had accumulation indices of 1.79 (1.25–2.77) and 1.84 (1.43–2.88), respectively. No significant laboratory changes were attributed to multiple dose drug administration. Local subcutaneous vasculitis and panniculitis were reported in rabbits receiving HCB. Ante-mortem examination did not find clinical disease, however 6 of 8 rabbits, including controls, had multifocal inflammatory changes in the liver and lungs on histopathology. Subcutaneous high concentration buprenorphine in rabbits has substantial pharmacokinetic variability and accumulation of both the parent drug and metabolites occurs with multiple dosing regimens. Subclinical illness may have impacted these findings and is a notable study limitation. Local inflammation in subcutaneous fat at injection sites was noted in rabbits administered HCB. High concentration buprenorphine has a quick onset of action when administered subcutaneously in the rabbit, but individual variability is notable. Subclinical inflammatory illness may prolong buprenorphine metabolism and clearance and should be considered in determining dosing regimens in rabbits, especially if disease is present or suspected. Inflammation at HCB injection sites is a drug sequela; monitoring injection sites is recommended.

GP2017-HCF, a high concentration formulation, demonstrates similar pharmacokinetics, immunogenicity and safety to GP2017, an approved adalimumab biosimilar

ABSTRACT Background GP2017 is an adalimumab biosimilar. The objective of this study is to compare the pharmacokinetics (PK) of GP2017 in its approved formulation and GP2017-high concentration formulation (HCF) in a randomized, double-blind, two-arm PK bridging study. Research design and methods Healthy male subjects received a single 40 mg subcutaneous injection of either GP2017-HCF (n = 162) or GP2017 (n = 168). PK, safety, and immunogenicity were assessed over 72 days post-injection. Results The 90% confidence intervals [CIs] of geometric mean ratios between GP2017-HCF and GP2017 for Cmax, AUC0-inf, AUC0-360 and AUC0-last were within the pre-defined margin of 0.80 to 1.25; thus, PK comparability between GP2017-HCF and GP2017 was demonstrated. Subgroup analysis of PK comparability by anti-drug antibody (ADA) subpopulation showed that the 90% CIs of geometric mean ratios between GP2017-HCF and GP2017 for Cmax, AUC0-inf, AUC0-360 and AUC0-last were within the margin of 0.80 to 1.25 in ADA-positive and ADA-negative subjects. The proportions of subjects with positive ADA responses and with neutralizing antibodies were comparable between the GP2017-HCF and GP2017 groups. GP2017-HCF and GP2017 were well tolerated, and there were no reports of deaths or other serious adverse events. Conclusion Results show PK comparability between GP2017-HCF and GP2017 and comparable safety and tolerability.