A phase I double-blind, placebo-controlled, safety, tolerability, pharmacokinetic and pharmacodynamic study of ascending single doses of ANF-Rho in comparison to standard of care-Neulasta® in healthy volunteers

ANF Rho is a new kind of pegfilgrastim which is similar to Neulasta®. Neulasta®
is a drug registered for treatment of a shortage of white blood cells in order
to prevent infections. It is used mostly for cancer patients to treat the side
effects of chemotherapy. Neulasta® is the brand name; the active ingredient is
called pegfilgrastim. This is a protein which is very similar to the human
version of the growth factor *granulocyte colony stimulating factor* (also
known as G-CSF or filgrastim). G-CSF is present in the human body by nature.
Therefore, Neulasta® is called a *biological*. The difference between naturally
occurring G-CSF and pegfilgrastim is the attachment of a large chain of
molecules (a polymer) to the protein. This makes the protein stay longer in the
body so patients need to receive drug less often to achieve the same effect.
ANF Rho is similar to Neulasta® but contains somewhat more polymer. Both
Neulasta® and ANF Rho are produced with the help of bacteria which have
received a human gene which makes them able to produce this protein. This is
the first time that the ANF Rho compound is being given to humans.

The purpose of the study is research.

The study investigates to what extent ANF Rho is tolerated in comparison to
Neulasta® (representing standard of care treatment) and placebo. A placebo is
the same formulation as the study medication without the active ingredient. It
will also be investigated how quickly and to what extent ANF Rho is absorbed
and eliminated from the body (this is called pharmacokinetics). In addition,
the effect of the compound on blood cells and bone will be investigated (this
is called pharmacodynamics).

This study will be performed in maximally 93 male and/or female volunteers
divided over 9 groups of 5 to 12 participants each.

The actual study will consist of 1 period during which you will stay in the
clinical research center in Zuidlaren for 8 days (7 nights) followed by 14 days
during which you will visit the clinical research center in Zuidlaren on 4
occasions (on Day 8, Day 10, Day 12, Day 14). The follow-up visit will take
place on Day 22. Your participation to the entire study, from pre-study
screening until the post study screening, will be maximally 7 weeks.

On Day 1 you will receive ANF Rho, Neulasta® or placebo after breakfast in the
morning through an injection under the skin of the abdomen. In the highest dose
group (50 µg/kg) of ANF Rho, the dose will be split into 2 subcutaneous
injections. The injection volume will be less than 1 mL per injection.

All potential drugs cause side effects; the extent to which this occurs
differs. The overall risks of ANF Rho administration are considered to be
minimal, although some are unforeseeable as the testing of this drug is still
at an early stage. As ANF Rho will be administered to humans in this study for
the first time, side effects in humans have not been reported to date. With the
dose used in this study no serious side effects are expected, but as all drugs
may potentially cause side effects to some extent, the occurrence of known or
other effects cannot be excluded. This means that there is a chance of a minor
side effect and a remote chance of something serious happening. It is expected
that ANF Rho, which resembles Neulasta®, will likely have the same side effects
as Neulasta®.

Neulasta® has been used now for 12 years and has been registered as a drug in
over 100 countries worldwide. The most common side effects of Neulasta® (seen
in more than 1 in 10 patients) are bone and muscle pain, headache and nausea
(feeling sick). Other common side effects (seen in between 1 in 10 and 1 in 100
patients) are reactions where the injection is given such as pain. Less common
but potentially serious reactions can include a low number of blood platelets,
allergic reactions, rapid drop in blood pressure, enlarged or ruptured spleen,
skin reactions and lung problems.

The body may recognize ANF Rho as foreign. As a result an immune response can
occur, for example by making antibodies against the study medication. However,
based on experience with Neulasta®, the chance of this happening is considered
very low.