Pharmacokinetics of GLP2-2G-XTEN in cynomolgous monkeys.

<p>GLP2-2G-XTEN plasma concentration is shown following 25 nmol/kg administration via intravenous (triangle) or subcutaneous (square) route. Three animals were dosed by each route of administration. Data points are the average ± s.d. of three animals per time point.</p

GLP2-2G-XTEN administration improves small intestine histopathology in the indomethacin-induced rat disease model.

<p>A) Hematoxylin and eosin staining of jejunum sections from A) diseased, vehicle-treated rats, B) diseased, GLP2-2G-XTEN high dose treated rats (125 nmol/kg total dose divided into five daily 25 nmol/kg doses), C) diseased, GLP2-2G-XTEN low dose treated rats (75 nmol/kg total dose given once on day −3) and D) diseased, GLP2-2G-XTEN low dose treated rats (divided into three 25 nmol/kg doses on day −3, −1 and 1).</p

Efficacy of low dose GLP2-2G-XTEN treatment in rat indomethacin-induced disease model.

<p>Data shown are effects on A) small intestine adhesions and B) small intestine trans-ulcerations. Treatment with GLP2-2G peptide (12.5 nmol/kg per injection; twice per day for 5 days) or GLP2-2G-XTEN (25 nmol/kg once daily or only once at the indicated doses) was initiated on Day −3 and sacrifice is on Day 2. Data are means and SE, n = 10 per group. * P<0.05 compared to diseased, vehicle-treated rats.</p

Allometric scaling human projection based on mouse, rat and monkey pharmacokinetic data.

<p>Allometric scaling was performed by fitting a linear regression model with the log of animal mass as the X variable and log of terminal half-life as the Y variable. The resulting linear model was extrapolated to a 70 kg human to produce a predicted value. The terminal half-life in humans can also be estimated using the predicted values for clearance (Cl) and volume of distribution (Vd) in the formula T½ = 0.693×Vd/Cl. Applying this formula yields a predicted terminal half-life of 240 hours in humans, which agrees with the linear extrapolation.</p

Treatment effects on small intestine length correlate with small intestine histopathology.

<p>A) Small intestine length B) villi height and C) mucosal atrophy, ulceration, infiltration measurements from diseased, vehicle-treated, GLP2-2G peptide-treated, and GLP2-2G-XTEN-treated rats. Quantitative histopathology was not performed on GLP2-2G-XTEN 75 nmol/kg single dose group.</p

Efficacy of equal molar GLP2-2G-XTEN and GLP2-2G peptide in rat indomethacin-induced disease model.

<p>Data shown are A) change in body weight from day −3 through day 2, B) small intestine length, C) small intestine adhesion score, D) small intestine trans-ulceration score and E) small intestine TNFα concentration. Open bars are healthy rats treated with vehicle; colored bars are indomethacin-induced diseased rats treated with vehicle, GLP2-2G peptide (12.5 nmol/kg per injection; twice per day for 5 days) or GLP2-2G-XTEN (25 nmol/kg once per day for 5 days) as indicated. Compounds were dosed starting three days prior to first indomethacin injection (Day −3) and data shown are from the time of sacrifice on Day 2 (24 hours post second indomethacin injection). Data are means and SE, n = 10 per group. * P<0.05 compared to diseased, vehicle-treated rats, ^# P<0.05 compared to diseased GLP2-2G peptide treated rats.</p

Intestinotrophic effects of GLP2-2G peptide and GLP2-2G-XTEN.

<p>Small intestine weight A) and length B) in normal rats following treatment with a total dose of 300 nmol/kg GLP2-2G peptide or GLP2-2G-XTEN. Data shown are from day 12 following SC injection of GLP2-2G peptide (12.5 nmol/kg per injection; twice per day for 12 days) and GLP2-2G-XTEN (25 nmol/kg per injection; once per day for 12 days). Data are means and SE, n = 10 per group. * P<0.05 compared to vehicle-treated rats, ^# P<0.05 compared to GLP2-2G peptide treated rats.</p

<i>In vitro</i> characterization of purified GLP2-2G-XTEN protein.

<p>A) ESI-MS analysis, major peak 83,142 Da – full length intact GLP2-2G-XTEN; minor peak 83,003 Da – des-His GLP2-2G-XTEN. B) non-reducing SDS-PAGE. Lane 1: molecular weight markers, lanes 2–4: 2 µg, 5 µg and 10 µg of GLP2-2G-XTEN, respectively. C) Size exclusion HPLC analysis. Solid curve: GLP2-2G-XTEN, 20 µg; dashed curve: molecular weights standard, which includes thyroglobulin 670 kDa, IgG 156 kDa, BSA 66 kDa, ovalbumin 45 kDa, myoglobin 17 kDa. D) Potency GPCR assay of GLP2-2G EC50 = 7 nM (95% confidence interval 4 nM to 12 nM) and GLP2-2G-XTEN EC50 = 380 nM (95% confidence interval 330 nM to 430 nM).</p

Construct 1 inhibits increase in blood glucose after end of fasting in cynomolgus monkeys.

<p>The effect of long-lived construct 1 on appetite suppression was tested in normal cynomolgus monkeys. Panels A–C show overlaid plots of blood glucose profiles after placebo or construct 1 administration for each individual animal. Solid arrows mark the time when food was returned to the animals (t = 6 hours).</p

Gcg-XTEN confers temporally-controlled resistance to insulin-induced hypoglycemia in dogs.

<p>Beagle dogs were fed three hours prior to the start of the experiment and fasted thereafter. At time = 0, animals received either a dose of 0.6 nmol/kg Gcg-XTEN or placebo (open arrows). Animals (n = 4 per group) received a challenge of 0.05 U/kg insulin to induce hypoglycemia at either 6 hr (A) or 12 hr (B) after initial dose, indicated by solid arrows. Values shown are the average blood glucose plus or minus the standard deviation. (C) A hypothetical timeline for human administration. Assuming Gcg-XTEN dosing at 21:00, 6 hr post dose corresponds to 03:00 (during sleep) where protection of hypoglycemia is desired, and 12 hr post dose corresponds to 09:00 where the pharmacodynamic effect should have expired to allow for a morning meal.</p