A novel needle for subcutaneous injection of interferon beta-1a: effect on pain in volunteers and satisfaction in patients with multiple sclerosis.

BACKGROUND: To reduce injection pain and improve satisfaction, a thinner (29-gauge [29G]), sharper (5-bevel) needle than the 27G/3-bevel needle used previously to inject interferon (IFN) beta-1a, 44 or 22 mcg subcutaneously (sc) three times weekly (tiw), was developed for use in multiple sclerosis (MS). METHODS: Two clinical trials in healthy volunteers and five surveys of patients with MS were conducted to assess whether the 29G/5-bevel needle with a Thermo Plastic Elastomer (TPE) needle shield (a sleeve that houses the tip of the needle in a secure location) is an improvement over the 27G/3-bevel needle with a rubber shield for injection of IFN beta-1a, 44 or 22 mcg sc tiw. Parameters assessed were: pain and ease of insertion (healthy volunteer and nurse responses on subjective pain measurement scales); and patient satisfaction (surveys of patients with MS). RESULTS: In healthy volunteers, the 29G/5-bevel needle with TPE shield was associated with the least perceived pain on the Visual Analog Scale (VAS) and Verbal VAS (VB-VAS); mean VAS pain scores decreased by 40% and skin penetration improved by 69% compared with the 27G/3-bevel needle with standard rubber shield (p < 0.01). Pooled results from surveys of patients with MS indicated that 63% of patients thought that injections were less painful with the 29G/5-bevel needle than the 27G/3-bevel needle. Results from individual surveys indicated that the 29G/5-bevel needle was an improvement over the 27G/3-bevel needle for ease of insertion, injection-site reactions, bruising, burning and stinging. CONCLUSION: Together these studies indicate that the 29G/5-bevel needle with the TPE shield is an improvement over the 27G/3-bevel needle with standard rubber shield in terms of pain, ease of insertion and patient satisfaction. These improvements are expected to result in improved compliance in patients with MS treated with IFN beta-1a, 44 or 22 mcg sc tiw.

The Rebif new formulation story: it's not trials and error.

The rapid evolution of the biopharmaceutical industry and the development of innovative technologies have provided an opportunity to improve recombinant interferon (IFN)-beta formulations. A number of strategies have been developed to improve the stability, tolerability and immunogenicity of IFNbeta formulations that are used in the long-term treatment of patients with multiple sclerosis (MS). This review focuses on the production of recombinant IFNs and discusses the development of one such biopharmaceutical, Rebif New Formulation (RNF).RNF was developed with the aim of further improving the tolerability and immunogenicity of Rebif, an approved IFNbeta-1a formulation administered subcutaneously three times per week (sc tiw). To this end, numerous candidate drug vehicles and formulations were developed. However, unlike other formulations of IFNbeta, the new candidate formulations in this case were free from all serum-derived components. Specifically, each RNF candidate was free from human serum albumin and produced without fetal bovine serum. The physicochemical stability, injection-site tolerability, pharmacokinetic profile and immunogenic potential of each candidate formulation were systematically tested. This involved initial screening of a large pool of formulations for promising candidates. Two candidate formulations were selected and subjected to further, extensive evaluation.Ex vivo T-cell assays were used to compare the immunogenicity of RNF candidates with that of the current (at the time of writing) approved formulation and an IFNbeta standard. A single RNF candidate induced less T-cell activation, in terms of proliferation and proinflammatory cytokine secretion, than the other two formulations. The results provided ex vivo evidence of the improved immunogenic potential of RNF. A murine model was used to compare the relative immunogenicity of RNF in vivo with two approved formulations of IFNbeta-1a. Mice treated with RNF developed neutralising antibodies more slowly and produced lower titres than mice treated with equivalent doses of the current IFNbeta-1a sc tiw formulation or another approved IFNbeta-1a formulation administered intramuscularly once per week (Avonex). RNF also demonstrated better local tolerability than the current IFNbeta-1a sc tiw formulation after single subcutaneous doses in healthy volunteers. One RNF candidate was superior to the others in all preclinical and phase I studies, and was chosen as the final RNF. This formulation is currently undergoing assessment in a 96-week, phase IIIb clinical trial in patients with MS. This single-arm, open-label, multicentre study will compare the immunogenicity and tolerability of RNF with historical data on the current formulation; results of a 48-week, interim analysis indicate that RNF has improved local tolerability and immunogenicity compared with the current formulation. It is anticipated that the benefits of RNF will translate into an improved long-term benefit-to-risk profile. Further assessment of RNF and other MS drugs is ongoing with the aim of enhancing the therapeutic options available for patients with MS.

Immunogenicity comparison of interferon beta-1a preparations using the BALB/c mouse model: assessment of a new formulation for use in multiple sclerosis.

The in vivo immunogenicity of a new interferon (IFN) beta-1a product (Rebif New Formulation; RNF) was compared with that of two approved recombinant human IFN beta-1a products (Rebif and Avonex). Immunogenic potential was assessed based on time to development of neutralizing antibodies (NAbs) and NAb titer. Female BALB/c mice (six in each group) received RNF, Rebif or Avonex (1.0 microg/mL subcutaneously three times weekly), and serum samples collected on Days 7, 21, and 35 (Study 1), or 28, 42, 49, and 60 (Study 2) were assayed for NAbs. In Study 1, no mice had NAbs at Day 7, but by Day 21 one mouse in the RNF group had NAbs, compared with three and four mice in the Rebif and Avonex groups, respectively. Results were similar in Study 2. All control mice were NAb negative; all actively treated mice had NAbs by day 35 or 42. Throughout Study 1, NAb titers were lowest in the RNF group and highest in the Avonex group, and at day 35, NAb titers were significantly lower in the RNF group than the Rebif group (p = 0.037). Results indicate that, on a gram-for-gram basis, RNF appears less immunogenic than Rebif or Avonex.

Immunogenicity and tolerability of an investigational formulation of interferon-beta1a: 24- and 48-week interim analyses of a 2-year, single-arm, historically controlled, phase IIIb study in adults with multiple sclerosis.

BACKGROUND: RNF (Rebif New Formulation, Merck Serono International S.A., Geneva, Switzerland), a formulation of interferon-beta1a (IFN-beta1a) without human- or animal-derived components, is currently under investigation. It was developed with the aim of maximizing the treatment benefit for patients with multiple sclerosis (MS) by improving injection tolerability and reducing the development of neutralizing antibodies (NAbs). OBJECTIVE: This paper reports the results of planned 24- and 48-week interim analyses comparing immunogenicity and tolerability data from an ongoing study of RNF with historical-control data for the currently approved formulation of IFN-beta1a from the EVIDENCE (EVidence of Interferon Dose-response: European North American Comparative Efficacy) study. METHODS: Patients in the 96-week, multicenter, singlearm, Phase IIIb RNF study received 44 microg/0.5 mL SC tiw; patients in the EVIDENCE study received an identical regimen of the currently approved formulation of IFN-beta1a. Criteria for inclusion in the RNF study were age between 18 and 60 years, an Expanded Disability Status Scale (EDSS) score <6.0, and a diagnosis of relapsing MS (McDonald criteria). Criteria for inclusion in the EVIDENCE study were age between 18 and 55 years, an EDSS score of 0 to 5.5, and a diagnosis of clinically definite relapsing-remitting MS (Poser criteria). Patients in both studies were treatment naive. Both studies used the same cytopathic-effect assay for NAbs to assess immunogenicity; patients who had NAb titers >or=20 neutralizing units (NU)/mL were considered NAb+. The primary end point was to compare the proportions of NAb+ patients in the RNF study and the historical data. Comparisons were descriptive and used exact 95% CIs. Safety analyses included 8 prespecified adverse events (AEs) of interest. RESULTS: Baseline demographic characteristics were well balanced between the RNF (N = 260) and EVIDENCE (N = 339) studies, except that patients in the RNF study were slightly younger (median age, 34.0 vs 39.0 years, respectively), and a few had secondary progressive MS (n = 6) or progressive relapsing MS (n = 1). At week 48, 87.3% of patients in the RNF study remained on treatment. The incidence of the prespecified AEs of interest in the RNF and EVIDENCE studies was as follows: flu-like symptoms (70.8% and 48.1%, respectively), injection-site reactions (29.6% and 83.8%), hepatic disorders (13.1% and 16.8%), cytopenia (9.6% and 11.8%), depression and suicidal ideation (5.8% and 19.8%), skin rashes (5.4% and 12.1%), hypersensitivity reactions (5.4% and 3.2%), and thyroid disorders (2.3% and 5.0%). Overall, the majority (96.9%) of AEs in the RNF study were mild (69.5%) or moderate (27.5%) in severity. The proportions of patients in the RNF and EVIDENCE studies with NAbs at both 24 and 48 weeks were 2.5% (95% CI, 0.9-5.5) and 14.3% (95% CI, 10.7-18.6), respectively; the proportions with NAbs at week 48 only were 13.9% (95% CI, 9.9-18.7) and 24.4% (95% CI, 19.9-29.4). The proportions of NAb+ patients with high NAb titers (>1000 NU/mL) at week 48 were 11.1% in the RNF study and 19.5% in the EVIDENCE study. CONCLUSIONS: The results of these interim analyses suggest that RNF had an improved overall tolerability and safety profile and a lower immunogenic potential compared with the approved IFN-beta1a formulation assessed in the EVIDENCE study. Two-year results from the RNF study are anticipated before the end of 2007.

Assessment of the immunogenicity of different interferon beta-1a formulations using ex vivo T-cell assays.

Recombinant homologues of human proteins have the potential to induce an immunogenic response when used therapeutically. Each of the three interferon (IFN) beta therapies currently approved for multiple sclerosis can induce the development of neutralising antibodies, the full effects of which on IFN beta therapy remain unclear. To investigate the immunogenicity of the currently licensed formulation of subcutaneous IFN beta-1a, 22 or 44 microg three times weekly (Rebif), two new formulations of IFN beta-1a (Rebif) New Formulation [RNF]1 and RNF2) have been developed. In this study, the immunogenicity of the current formulation was investigated against RNF1, RNF2 and an IFN beta standard using ex vivo T-cells. Dendritic cells, isolated from peripheral blood monocytes donated by 26 healthy volunteers, were matured in vitro and incubated with the test antigens for 6h. Autologous CD4(+) T-cells from the same donors were added and further incubated before cytokine release was assessed by ELISpot assay and proliferation by [(3)H]thymidine pulse. Secretion of T-cell-derived interleukin-2 was 79%, 66% and 105% in incubations with RNF1, RNF2 and the current formulation, respectively (normalised to secretion with the IFN standard; p<0.05, RNF2 versus the current formulation). Secretion of IFN gamma was highly variable between donors, with no significant difference observed between formulations. Normalised values for T-cell proliferation were 56%, 44% and 88% with RNF1, RNF2 and the current formulation, respectively (p<0.05, RNF2 versus the current formulation). The immunogenic potential of RNF2 is significantly lower than that of the current formulation when tested ex vivo. This result is now being confirmed through ongoing clinical trials.

A comparison of the biologic activity of two recombinant IFN-beta preparations used in the treatment of relapsing-remitting multiple sclerosis.

Recent clinical trials with interferon-beta (IFN-beta) in relapsing-remitting multiple sclerosis (RRMS) have clearly demonstrated that the IFN-beta dosing regimen affects the clinical efficacy, thereby highlighting the importance of determining the relative biologic activities of the IFN-beta products currently available. Although studies have been published that examine the biologic activities of the two structurally different forms of recombinant IFN-beta, IFN-beta1a (Rebif), Serono, Geneva, Switzerland) and IFN-beta1b (Betaseron)/Betaferon), Berlex [Montville, NJ]/Schering [Berlin, Germany]), there have been few direct comparative studies. Therefore, to obtain a more accurate estimate of the relative biologic activities of Rebif and Betaseron, this study examined the antiviral activities of these two products within the same assay system and against the same natural human IFN-beta standard. Whereas the manufacturers' information suggests that the bioactivity of Betaseron is only about 8.7-fold less than that of Rebif, the results of the present direct, comparative study show that Rebif has an antiviral activity 14 times greater than that of Betaseron. This may have important clinical implications, because on the basis of the results reported here, Rebif at 44 microg t.i.w. is approximately double the maximal licensed weekly dose for Betaseron.