Aug 9, 2002 - tration of the dipeptidyl peptidase IV inhibitor isoleu cine thiazolidide. .... It has been further surprisingly discovered that the chronic oral ...
US 20030119736A1 US 20030119736A1
(19) United States (12) P a ten t A p p lica tio n P u b lication (io) Pub. No.: U S 2003/0119736 A l Demuth et al. (43) Pub. Date: Ju n . 26,2003 (54)
(76)
METHODS FOR IM PROVING ISLET STGNAT JN G IN DIABETES M ELLITUS AND FOR ITS PREVENTION Inventors: Hans-Ulrich Demuth, Halle/Saale (DE); Konrad Glund, Halle/Saale (DE); J. Andrew Pospisilik, West Vancouver (CA); Kerstin Kuehn-W ache, Halle/Saale (DE) Correspondence Address: BROW N, RUDNICK, BERLACK & ISRAELS, LLP. BOX IP, 18TH FLOOR ONE FINANCIAL CENTER BOSTON, M A 02111 (US)
(21)
A ppl. No.:
10/216,349
(22)
Filed:
A ug. 9, 2002 Related U.S. Application Data
(63)
Continuation-in-part o f application No. 09/824,622, filed on Apr. 2, 2001, now Pat. No. 6,500,804.
Publication Classification (51) (52)
(57)
Int. Cl.7 ...........................A 61K 38/18; A 61K 31/70 U.S. C l..................................................... 514/12; 514/23
ABSTRACT
The present invention discloses methods for therapeutically treating mammals, including but not limited to humans, to increase the relative insulin producing performance of endogenous pancreatic β-cells, to cause differentiation of pancreatic epithelial cells into insulin producing β-cells, to improve muscle sensitivity to insulin and other weight control efforts by the chronic oral administration o f a DP IV-inhibitor. The administration causes the active form of G LP-I and other non-nutrient stimulated growth hormones to remain biologically active longer under physiological conditions. The extended presence o f such hormones, in particular in the pancreatic tissue can also facilitate differ entiation and regeneration of the β-cells already present that are in need of repair.
Patent Application Publication
Fig. I
GLP-Ia 25 η
c
ο 15 c φ
10
O
C
-
O O
O
50
100
150
tim e (min)
200
250
300
US 2003/0119736 A l
-50
Sheet I of 16
"7,5 mg ■15 mg — A- 30 mg - -K- 60 mg — X—-120 mg -240 mg —I—-Placebo P01 -Placebo P02
-
Jun. 26,2003
20
Patent Application Publication
Jun. 26,2003
Sheet 2 of 16
US 2003/0119736 A l
Fig. 2
GLP-Ia (AUC 250min) v s P32/98
C O O
A la » S e r, T h r» G ly , Val. W hile the proteins D P IV, DP II, FA Pa (Seprase), DP 6, DP 8 and DP 9 are structurally related and show a high sequence homology, attractin is an extraordinary functional DP IV-Iike enzyme (49). [0056] Further DP IV-Iike enzymes are disclosed in WO 01/19866, WO 02/04610, WO 02/34900 and WO02/31134. WO 01/19866 discloses human dipeptidyl aminopeptidase 8 (DPP8) with structural und functional similarities to DP IV and fibroblast activation protein (FAP). The dipeptidyl pep tidase IV-Iike enzyme o f WO 02/04610 is well known in the art. In the GENE BANK data base, this enzyme is registered as KIAA1492 (registration in February 2001, submitted on Apr. 4, 2000, AB040925) and in the MEROPS data base. WO 02/34900 discloses a dipeptidyl peptidase 9 (DPP9) with significant homology to the amino acid sequences of DP IV and DPP8. W O 02/31134 discloses three DP IV-Iike enzymes, DPRP1, DPRP2 and DPRP3. Sequence analysis revealed that D P R P l is identical to DPP8, as disclosed in WO 01/19866, that DPRP2 is identical to DPP9 and that DPRP3 is identical to KIAA1492 as disclosed in WO 02/04610. [0057] A s a consequence of the resulting enhanced stabil ity of the endogenous GLP-1 (including G LP-l-derived) circulating peptides caused by the inhibition of DP IVactivity, GLP-1 activity is prolonged resulting in function ally active GLP-1 (including G LP-l-derived) circulating peptide hormones facilitating growth-hormone-like stim u lation of pancreatic cells in such a way that these cells proliferate to functionally active cells of the Islets of Langerhans. Additionally, insensitive pancreatic cells or impaired pancreatic cells may be transformed into functionally active cells of the islets of Langerhans when exposed to GLP-1. [0058] It w as expected, that the transformation of insen sitive pancreatic cells or impaired pancreatic cells to func tionally active cells of the islets of Langerhans results in an increased insulin secretion and in an increased insulin level in blood plasma. Surprisingly, in studies in healthy human volunteers and obese, diabetic Zucker rats, the insulin level decreased after treatment with the DP IV-inhibitor isoleucyl thiazolidine hemifumarate (P32/98) (see examples I and 2, respectively). Nevertheless, the resulting regeneration of the islets of Langerhans does change the efficacy of endogenous
US 2003/0119736 A l
Jun. 26, 2003 4
insulin and other islet hormones, such as glucagon, in such a w ay that stimulation of carbohydrate metabolism o f a treated mam mal is effected. As a result, the blood glucose level drops below the glucose concentration characteristic for hyperglycemia, as shown in examples I and 2. The mechanism triggering these effects is not known in detail. However, this resulting regeneration o f the islet cells further affects anomalies of the metabolism including glucosuria, hyperlipidaemia as well as severe metabolic acidosis and Diabetes mellitus, by preventing or alleviating these sequelae. It has been further surprisingly discovered that the chronic oral administration of effectors of DP IV, such as orally active inhibitors thereof, can also result in increased rate o f progression to satiety during nutrient ingestion, decreased weight, decreases in chronic or long-term weight gains, and improved insulin sensitivity in muscles. Still another unforeseen effect involves the increased availability of hormones which are not regulated primarily in the short term by nutrient ingestion (e.g. acute changes in glucose levels) and which can improve β-cell activity and/or increas ing differentiation o f pancreatic cells to β-cells resulting in m easurably improved insulin output. [0059] In contrast to other proposed methods known in the art, such as pancreatic cell or tissue transplantation or ex-vivo treatment of pancreatic cells using GLP-1 or exendin-4 followed by re-implantation o f the treated cells, the present invention does not cause or require complicated and costly surgery, and provides an orally available therapy. The instant invention represents a novel approach for lowering the elevated concentration o f blood glucose, modifying satiety, weight gain, muscle sensitivity amongst other related effects. It is commercially useful and suitable for use in a therapeutic regime, especially concerning human disease, many of w hich are caused by prolonged elevated or blood glucose levels or improper or inadequate β-cell activity. BRIEF DESCRIPTION OF THE FIGURES [0060] Further understanding o f the instant invention may be had by reference to the figures wherein: [0061] FIG. I is a graphical representation o f the timedependency of circulating bioactive GLP-1 in humans (n=36) depending on the orally applied DP IV-inhibitor formulation P32/98; [0062] FIG. 2 is a graph representing the dependency of the AUC o f circulating bioactive GLP-1 in humans (n=36) on the orally applied DP IV-inhibitor formulation P32/98; [0063] FIG. 3 is a graphical representation showing the improvement of morning blood-glucose (MBG) after sub chronic mono therapeutic application o f 8.7 mg/kg/d of P32/98 to obese, diabetic fa/fa rats; [0064] FIG. 4a is a graphical representation showing improved glucose-control due to DP IV-inhibitor treatment after 16-days of treatment in obese diabetic rats [0065] FIG. 4b is a graphical representation showing reduced insulin-secretion due to DP IV-inhibitor treatment after 16 days of treatment in obese diabetic rats; [0066] FIG. 5a is a graphical representation showing the blood glucose levels as a function of time in the maintenance of improved glycemia after 21 days o f subchronic treatment of obese, diabetic fa/fa rats by the formulated DP IVinhibitor P32/98;
[0067] FIG. 5b is a graphical representation showing the plasma insulin levels as a function o f time in the m ainte nance of improved glycemia after 21 days of sub-chronic treatment of obese, diabetic fa/fa rats by the formulated DP IV-inhibitor P32/98; [0068] FIG. 6 shows body w eight and water intake m ea sured in DP IV-inhibitor treated (open circles) or control (solid squares; n=6) VDF rats. Body weight (A), and water intake (B) were measured along with morning and evening blood glucose levels and food intake (not shown) every two days. Statistical significance (p
