A Randomized, Double-blinded, Active Controlled Crossover Clinical Trial to Investigate PK, PD and Safety of GBPD002
Condition: Natural Blood and Blood Product Toxicity
Study type: Interventional
Phase: Phase 1/Phase 2

Summary: 
      Erythropoietin (EPO) biosimilarity for GBPD002 (test candidate) and Eprex® (comparator) has
      been evaluated by comparing the pharmacokinetic (PK) and pharmacodynamic (PD) properties
      following subcutaneous injection in human subjects. This was a randomized, double-blind,
      two-sequence, crossover study. Subjects were randomly assigned and received a dose (4,000 IU)
      of either the test or comparator EPO. The subjects received the alternative formulation after
      the wash out period (4 weeks) of the first administration. The primary PK parameters, viz.,
      maximum observed concentration (Cmax) and area under the curve extrapolated to infinity
      (AUC00-inf), were calculated with the serum EPO concentrations from blood samples and were
      found comparable for both formulations. The geometric mean ratios (@90% CI) of the Cmax and
      AUCinf were 1.16 and 0.89, respectively, which were within the regulatory range of 0.80-1.25.
      The reticulocyte, hematocrit, hemoglobin and red blood cell counts were measured as PD
      markers. The time-matched serum EPO concentrations and PD markers denoted a counterclockwise
      hysteresis, and thereby suggesting a time delay between the observed concentration and the
      response. ANOVA derived P-values (all were greater than 0.05) for the effectors clearly
      revealed the similarity between effects on PD markers for both formulations. Both
      formulations were found tolerated well, and anti-drug antibodies were not observed. Thus, the
      two formulations are projected to be used interchangeably in clinical settings.
    

Detailed description: 
      Erythropoietin (EPO) is a glycoprotein hormone that plays a key role in the formation of red
      blood cells (RBCs). EPO is primarily synthesized in the peritubular cells of the kidney and
      released into the systemic circulation in adult individuals. Circulating EPO binds to the EPO
      receptor on bone marrow erythroid progenitors, triggering multiple signaling pathways that
      support differentiation into mature RBCs. A reduction in EPO production is the primary cause
      of anemia in people with chronic renal failure [4]. Human recombinant epoetin (rHuEPO) or
      erythropoiesis stimulating agents (ESA) have been demonstrated to stimulate erythropoiesis in
      anemic patients with chronic renal failure, including those who need and don't need dialysis.
      ESAs are used to treat chemotherapy-induced anemia in cancer patients and to reduce the
      requirement for allogenic blood transfusions in patients with mild anemia who are undergoing
      surgery. Furthermore, human recombinant epoetin is recommended for patients who are at high
      risk for perioperative transfusions due to considerable blood loss.

      Recombinant rHuEPO was manufactured using recombinant DNA techniques. Alpha epoetins are the
      most commonly used type of rHuEPO among the other forms. Eprex®, the pioneer product of alpha
      epoetins, is a regular medicine with proven efficacy and tolerability. Globe Biotech Limited,
      a Bangladeshi biopharmaceutical company has developed GBPD002, a biosimilar of Eprex® which
      is synthesized in genetically engineered Chinese hamster ovary (CHO) cells. Upstream and
      downstream process development and validation was done for large scale production. Step by
      step identical analytical results confirmed the biosimilarity of GBP002 with Eprex®. Single
      and repeat dose toxicity was performed in Wister rat to analyze the toxicity of GBPD002 with
      Eprex®.

      The aim of this study is to analyze the bioequivalence of GBPD002 and Eprex®. The purpose of
      this study is to compare the pharmacokinetic (PK), pharmacodynamics (PD) and safety of human
      recombinant erythropoietin, GBPD002, developed by Globe Biotech Limited and the reference
      product Eprex® manufactured by Janssen Cilag Ltd., UK in healthy volunteers. An independent
      ethics committee gave ethical clearance for this study and the protocol for this study was
      approved by the Directorate General of Drug Administration (DGDA) of Bangladesh.

      A randomized, double-blind, single-dose, and two-sequence crossover trial was conducted in 42
      healthy volunteers in 2 groups/cohorts/arms. The Clinical Research Organization (CRO Ltd.)
      conducted this investigation at Farabi General Hospital, Dhanmondi R/A, Dhaka 1209, in
      compliance with the principles of the Declaration of Helsinki and the International
      Conference on Harmonization's Guideline for Good Clinical Practice. Bangladesh's Directorate
      General of Drug Administration (DGDA) approved the study methodology and informed consent
      form. Volunteers were given thorough information about the study and they have signed an
      informed consent form to affirm their willingness to participate.

      The study was open to healthy male participants aged 19-45 years old who weighed 55.0-90.0 kg
      and had a body mass index of 18.0-27.0 kg/m2. Subjects were eliminated if they had at least
      one of the following clinical laboratory test results: Hemoglobin level <12 g/dL or >17 g/dL,
      vitamin B12 level <200 pg/mL, ferritin level <21.8 ng/mL, transferrin level <190 mg/dL and
      any anomalous range for the reticulocyte (RET) count, erythrocytes, platelets or serum
      potassium levels. The number of subjects was determined based on the results of a previous
      clinical study on a single subcutaneous injection of epoetin alfa in healthy subjects. The
      total number of subjects was 42, assuming a 20% dropout rate. COVID-19, HIV, HBsAg, and HCV
      (Hepatitis C Virus) positive individuals were excluded from the study.The recruited
      volunteers were randomly assigned to one of the two sequences and received a single
      subcutaneous injection of 4,000 IU of either the comparator drug (Eprex®) or the test drug
      (GBPD002) in the abdomen from a prefilled syringe (PFS), based on the allocated sequences
      with a 28-day washout period: sequence A, administered the comparator drug in period 1
      followed by the test drug in period 2; sequence B, given the test drug in period 1 followed
      by the comparator drug in period 2.Blood samples were taken for the PK evaluation at predose
      and at 1, 3, 6, 8, 10, 14, 24, 48, 72, 96, 120 and 144 h postdose. For the PD evaluation, the
      reticulocyte count (RET; %), hematocrit (HCT; %), haemoglobin (HB) (g/L) and red blood cell
      (RBC) count (106/mm3) were calculated at predose and at 72, 144, 216, and 312 h postdose.
    

Intervention name(s): Erythropoietin alpha
Intervention type(s): Biological
Intervention description(s): To analyze bio-similarity of GBPD002 with Eprex®, the Globe Biotech Ltd performed quite a few experiments to determine total analytical study of GBPD002. They performed peptide mapping to determine amino acid composition of GBPD002 and found similar result with erythropoietin alpha. The molecular weight was determined through SDS-PAGE analysis of GBPD002 and was compared with Eprex. The western blot analysis was performed to determine immune chemical characterization. Characterization of different isoforms was conducted through 2D gel electrophoresis.
Intervention MeSH terms: Epoetin Alfa
Allocation: Randomized
Intervention model: Crossover Assignment
Intervention model description: Healthy volunteers participating in two cohorts. In each cohort 21 volunteers are participating in the clinical trial.
Masking: Quadruple (Participant, Care Provider, Investigator, Outcomes Assessor)
Primary purpose: Treatment
Keywords: human recombinant erythropoietin