A randomized, double-blind, placebo-controlled PK/PD guided, phase I study in healthy volunteers given escalating, single intravenous doses of NI-0101 in the absence or presence of a systemic LPS challenge.

NI-0101 is a humanized immunoglobulin gamma (IgG)1 kappa (*) monoclonal
antibody (mAb) that binds and antagonizes TLR4. NI-0101 binds with high
affinity (dissociation constant, Kd = 139 pM), independently of MD-2 (myeloid
differentiation factor-2). NI-0101 prevents TLR4 dimerization and, thus, ligand
driven receptor-mediated intracellular signalling. Two amino acid
substitutions, introduced within the SKAF motif of the Fc-domain of NI-0101,
destroy the binding sites for interactions with Fc*RIII and the complement
component, C1q, while maintaining the binding to Fc*RI and to Fc*RII.
Polymorphisms for the Fc*RIIa will affect the potency of NI-0101 for blocking
TLR4-mediated cytokine release (half maximum inhibitory concentrations, IC50s
of 128, 223, 2191 pM for genotype 131R/R, 131R/H and 131H/H, respectively, with
distribution of the genotypes of 21%, 46%, and 33%, respectively).
Using NI-0101 and a murine NI-0101 surrogate (5E3), the role of TLR4 activation
in the pathogenesis and progression of RA has been further corroborated in a
number of experiments performed by NovImmune. Examples of these are the in vivo
experiments in murine animal models of RA (i.e. collagen-induced arthritis, CIA
and IL-1Rn-/- models) using 5E3 and the studies to investigate the effect of
NI-0101 in blocking the spontaneous production of inflammatory cytokines by
synovial tissue explants obtained from RA patients.
NI-0101 provides a novel and unique means of inhibiting the TLR4-mediated
inflammatory response and thus appears to be an attractive agent to blunt the
inflammatory response in diseases like RA, where inflammation initiated by TLR4
plays a critical role in the pathophysiology of the disease.

Part 1:
- To assess tolerability, safety and PK of escalating single Intravenous (IV)
doses of NI-0101 in healthy volunteers with the aim of covering a wide range of
plasma concentrations up to those theoretically reflecting a potential
therapeutic effect on inflamed tissues;
- To determine the PD effects of NI-0101 by an ex vivo whole blood challenge
with an exogenous TLR4 ligand (LPS) and, optionally, an endogenous ligand, as a
proof of mechanism;
- To determine the potential impact of Fc*RIIA polymorphism on NI-0101 effects;
- To investigate immunogenicity of NI-0101, i.e. the presence of anti-drug
antibodies (ADAs).

Part 2:
- To assess the tolerability, safety , pharmacodynamic effects and PK of
escalating single IV doses of NI-0101 in healthy volunteers exposed to an in
vivo LPS challenge;
- To describe the dose-concentration-inhibition relationship of in vivo and ex
vivo LPS challenges performed at the end of NI-0101 infusions;
- To demonstrate that the above described relationships hold with time after
NI-0101 administration;
- To investigate immunogenicity of NI-0101, i.e. the presence of anti-drug
antibodies.

Randomized, double-blind, placebo-controlled, PK/PD guided, single ascending
dose interventional Phase I study.

Part 1: assessment of effects using whole blood ex vivo stimulations (LPS and
optionally an endogenous ligand).
Part 2: assessment of effects using an in vivo challenge with LPS.

A single intravenous infusion of NI-0101 or placebo without (Part 1) or with
(Part 2) an in vivo LPS challenge.

NI-0101 is a humanized immunoglobulin gamma (IgG1) kappa monoclonal antibody
(mAb) that blocks both the signal transduction pathways of the trans-membrane
protein, human Toll-Like Receptor 4 (TLR-4). The matched placebo is a sterile
solution for intravenous infusion that is identical to the NI-0101 drug product
but does not include the active substance. LPS (lipopolysaccharide) is an
endotoxin frequently used in human research.

TLR4 signalling is essential in host defense and excessive and long-lasting
inhibition is associated with impaired host defense. Although there is a
theoretical chance that this may occur in the trial, it is unlikely as the
trial is designed so that excessive and long-lasting inhibition of TLR4
signalling is unlikely to occur. The dose of NI-0101 will not exceed a
predefined threshold of residual cytokine levels for adequate immunity against
microbial threat. It is considered that problems can be adequately managed in
case of serious adverse events calamity as CHDR is closely linked to Leiden
University Medical Center.
Determination of a safe starting dose was achieved by combining classical
methods and the MABEL approach using data on the in-vitro inhibition of the
compound assessed in human blood. Dose-escalation for dosing next cohorts will
only be performed after consensus by the Principle Investigator, the sponsor
and -if needed- external experts (Safety Review Committee) that this is safe to
proceed to the next dose level.