Open Label Single Period Pilot Clinical Trial to Standardize Procedures for Evaluation of Glucose Dependent Insulin Secretion in Subjects with Type 2 Diabetes Mellitus

In this study you will not participate in an investigation with a new compound,
but you will participate in a pilot study in which the reactions of patients
with Diabetes Mellitus Type 2 will be evaluated. In order to participate in
this study, you are not allowed to take your own diabetic medication for a
maximum of 1.5 days. In order to obtain a pre-defined blood sugar level, you
will receive a temporary insulin infusion administered with a pump. In Part 1
you will also receive an infusion of glucose solution. During this study you
will receive insulin. Insulin is an hormone produced by the human body
naturally. Insulin is involved in the regulation of the metabolism of sugar and
fat in the body. Insulin causes cells in the liver, muscle, and fat tissue to
take up blood sugar (glucose) from the blood, storing it as a long-term energy
depot in the liver and muscle. When the natural control of insulin production
by the body fails, the complication diabetes mellitus will develop. The insulin
that will be administered temporarily, is used on a large scale and is
internationally registered (NovoRapid) and approved.

To standardize the procedures and analyses to support evaluation of glucose
dependent insulin secretion in Type 2 Diabetes Mellitus in subjects with Type 2
Diabetes.

Design:
an open-label, single period pilot clinical study in a maximum of six type 2
diabetes mellitus patients to standardize procedures for evaluation of glucose
dependent insulin; each subject will receive a continuous intravenous infusion
of insulin for up to 12 hours starting on Day -1 of the study, followed by a
160-minute GGI procedure conducted on Day 1 of the study: glucose (20% glucose)
will be administered as a stepwise, graded infusion of 2, 4, 6, and 10
mg/kg/min (down-regulated to 8 mg/kg/min at last step if applicable), each
infusion step should be maintained over exactly 40 minutes, all subjects will
undergo a 10 hour overnight fast before initiation of the GGI. Of these 6
subjects, 4 subjects may be selected randomly to participate in Part 2 of the
study in which only the insulin infusion is given during the day.

Procedures and assessments
Screening and follow-up:
clinical laboratory, physical examination, weight, 12-lead ECG, vital signs
(including oral temperature), urine drug screen; at eligibility screening:
medical history, HbA1C, HBsAg, anti HCV, anti-HIV 1/2; weight and vital signs
to be repeated upon admission. For Part 2, no screening visit or follow-up
visit is necessary.

Observation period:
Part 1: one period in clinic from -24 h before start of the GGI up to
completion of the weaning procedure in the afternoon of Day 1
Part 2: one period in clinic from -12 h before start of the insulin infusion
up to completion of the insulin infusion the afternoon of Day 1

Blood sampling:
for glucose (Part 1 and Part 2): every 30 minutes during the insulin infusion.
for glucose (Part 1 only): 10 and 3 minutes before the start of the GGI, and
10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 120, 130, 140, 150 and 160 min after
start of GGI
for C-peptide (Part 1 only): 10 and 3 minutes before the start of the GGI, and
10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 120, 130, 140, 150 and 160 min after
start of GGI
for insulin (Part 1 only): 10 and 3 minutes before the start of the GGI, and
10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 120, 130, 140, 150 and 160 min after
start of GGI
for GLP-1/glucagon and for archive(Part 1 only): 10 minutes before the start of
the GGI and 40, 80, 120 and 160 min after start of GGI

Urine sampling:
for glucose (Part 1 only): interval from 15 min before start of GGI up to 160
min after start of GGI

Safety assessments:
adverse events: throughout the study; vital signs: 12 h and 60 min before GGI
and prior to discharge; blood sample for glucometer (Part 1) and iSTAT/YSI
(Part 2): every hour during insulin infusion; Only Part 2: 10 and 3 minutes
before GGI and 40, 80, 120 and 160 min after start of GGI and 10 times during
the weaning procedure

Insulin Infusion Procedures:
IV catheter placement: ~60 min before start of the insulin infusion ; insulin
infusion: 11 * 1 h before GGI (only Part 1), blood glucose concentration should
be kept around 5.6 mmol/L

GGI Procedure:
Part 1 only: infusion of glucose: 0 - 160 min on Day 1, glucose (20% glucose)
will be administered as a stepwise, graded infusion of 2, 4, 6, and 10
mg/kg/min (down-regulated to 8 mg/kg/min at last step if applicable), each
infusion step should be maintained over exactly 40 minutes

Bionalysis:
analysis of glucose samples using a clinical chemistry method by PRA
analysis of C-peptide samples using a clinical chemistry method by PRA
analysis of insulin samples using a clinical chemistry method by PRA
analysis of GLP-1/glucagon samples using a validated method by PRA

Insulin infusion

Procedures: pain, light bleeding, heamatoma, possibly an infection.