The ethical approval was taken from National Health Service Ethics Review Committee (NHERC) to conduct the trial and was carried with the Declaration of Helsinki and the principles of the International Conference on Harmonization which requires Good Clinical Practice.
Study Population
The study was carried out in Mirpur in Dhaka. The study recruited 41 healthy men and women. Participants had to weigh between 40.0 and 95 kg and have a BMI of 18.5–29.9 kg m− 2 to be eligible for the study. The study protocol required that all the vital signs of participants must be within normal limits, and evaluated by physical examination. Laboratory tests including blood and urinary parameter along with a 12-lead electrocardiogram (ECG) as well as a radio imaging of the chest must be within the normal range or declared clinically non-significant. Female participants must be tested negative for pregnancy (screened and confirmed by serum and urinary β-human Chorionic Gonadotropin/ β-hCG). We excluded participants having a history of malignancy, such as lymphoma, leukemia, or skin cancer. Study participants with a positive surface antigen test for hepatitis B, antibody test for hepatitis C, and/or IgG and/or IgM for core antibody test and positive antibody test for human immunodeficiency virus at screening were excluded from the study. Participants with a positive laboratory test for drug abuse by urine examination and a positive breath alcohol test were not enrolled. Smokers who smokes more than 10 cigarettes per day or refused to refrain from smoking or using any other product that contains nicotine throughout the trial's hospital stay were excluded. Other significant exclusion criteria were history and/or current presence of atopic allergy; any present or previous local fungal infections; active or latent tuberculosis; known or suspected hypersensitivity to any drugs; history of invasive systemic fungal infections; any infection that requires hospitalization and/or anti-infectives or antibiotics within 6 months prior to trial drug administration.
The participants were completely aware of the trial's goal and were given a concept of pharmacological effects as well as the potential adverse effects of the study drug. Written informed consent was obtained from all participants in accordance with the Declaration of Helsinki. In addition, each participant received a wage loss for taking part in the study.
Participants were screened around one week before receiving their adalimumab dose. Participants were admitted to the inpatient unit at the Universal Medical College and Hospital facilities 12 hours before dosing and stayed for two days following dosing. An interim safety analysis was undertaken with data accumulated at that point after 5 participants per arm had been examined for a minimum of 2 days.
Participants were randomly assigned to groups before receiving the investigational medicinal product (IMP). Single subcutaneous dose of 40 mg adalimumab; Advixa or Humira® was injected around 3 cm away from the navel or mid-thigh by trained nurses at the in-house clinical setting. Participants were required to remain in a relaxed state after receiving the medication and were not allowed to engage in intense activities. Blood samples were taken at each time point, and safety checks were performed as per the protocol.
Prior to dosing (-15 minutes) and at 4, 12, 24, 36, 48, 72 ± 2, 96 ± 2,120 ± 2, 144 ± 2, 168 ± 2, 192 ± 2, 360 ± 2, 528 ± 2, 696 ± 2, 864 ± 2, 1032 ± 2, 1200 ± 2, 1536 ± 2 hours blood samples were collected for pharmacokinetic analysis (19-time points).
All blood samples were collected in Serum Separator tubes. The samples were kept upright for 30 minutes at room temperature followed by centrifugation for 10 minutes at 2000 RCF (2°C-8°C). Serum samples were stored at or below − 70°C after aliquoting into duplicate microcentrifuge tubes.
PK Assessments And Endpoints
The concentrations of Advixa and Humira in the blood were measured by the ELISA method. Bound analyte, and 3,3′,5,5′-tetramethylbenzidine for colorimetric readout. were identified by using a TNF-coated plate, a horseradish peroxidase-conjugated anti-human IgG antibody. A plate reader was used to measure colorimetric intensity at 450 nm (detection) and 630 nm (reference) wavelengths.
Cmax, terminal half-life (t1/2), AUC from time 0 extrapolated to infinity [AUC (0)], and AUC from time 0 to the last quantifiable concentration [AUC (0, tlast)] were the primary PK endpoints.
Safety Evaluation
During the trial, all adverse events (AEs) reported by the participants during the study period were well documented. Any clinically significant aberration was reported as an AE when compared to the baseline. A safety check was done for subjects who completed the 1536-hour follow-up visit or withdrew from the trial early. When adverse events (AEs) occurred during the trial, the individual was tracked until the AE was resolved or stabilized. All adverse events were listed, including chest discomfort, pain, restlessness, headache, low blood pressure, swelling at the site of blood collection, vertigo, abdominal cramp, increased frequency of loose stool, upper abdominal burning sensation, itching at the site of IP administration, vomiting, burning sensation to the other thigh. The adverse events (AEs) associated with the study medication were summarized.
Statistical analysis
A one-way ANOVA model with treatment as a fixed effect was used to investigate AUC (0), AUC (0, tlast), and the (Cmax). The PK parameters were log-transformed for analysis, and the difference in mean parameters between the two groups was calculated, then equivalency was assessed by re-expressing on the original ratio scale. For each comparison, PK equivalence was declared if the 90 percent CI for the test: reference ratio fell entirely within the 70–145 percent equivalence margin. Bioequivalence was established if all three PK metrics met the PK equivalence criterion. Other PK metrics, serum drug concentrations, and safety/tolerance data were analyzed using descriptive statistics.