US 2011 0082079A1 (19) United States (12) Patent Application Publication (10) Pub. No.: US 2011/0082079 A1 Spetzler et al. (43) Pub. Date: Apr. 7, 2011 (54) GLUCAGON-LIKE PEPTDE-1 DERVATIVES Publication Classification AND THEIR PHARMACEUTICAL USE (51) Int. Cl. A638/22 (2006.01) (75) Inventors: Jane Spetzler, Bronsho (DK); C07K I4/575 (2006.01) Lauge Schäffer, Lyngby (DK); A6IP3/10 (2006.01) Jesper Lau, Farum (DK); Thomas A6IP 9/12 (2006.01) Kruse, Herlev (DK); Patrick A6IP3/04 (2006.01) William Garibay, Holte (DK); A6IP 9/10 (2006.01) Steffen Runge, Frederiksberg A6IPI/00 (2006.01) (DK); Henning Thogersen, Farum A6IPL/04 (2006.01) (DK); Ingrid Petersson, Frederiksberg (DK) (52) U.S. Cl. .......................... 514/7.2:530/399; 514/11.7 (57) ABSTRACT (73) Assignee: Novo Nordisk A/S, Bagsvard (DK) The invention relates to protracted Glucagon-Like Peptide-1 Appl. No.: (GLP-1) derivatives and therapeutic uses thereof. The GLP-1 (21) 12/676,451 derivative of the invention comprises a modified GLP-1 (7- PCT Fled: 37) sequence having a total of 2-12 amino acid modifications, (22) Sep. 5, 2008 including Glu22 and Arg26, and being derivatised with an PCT NO.: PCT/EP2008/061755 albumin binding residue or pegylated in position 18, 20, 23. (86) 30, 31, 34, 36, 37, or 39. These compounds are useful in the S371 (c)(1), treatment or prevention of diabetes type 2 and related dis (2), (4) Date: Oct. 13, 2010 eases. The compounds are potent, stable, have long half-lives, a high affinity of binding to albumin, and/or a high affinity of (30) Foreign Application Priority Data binding to the extracellular domain of the GLP-1 receptor (GLP-1R), all of which is of potential relevance for the overall Sep. 5, 2007 (EP) .................................. O7115746.5 aim of achieving long-acting, stable and active GLP-1 deriva Jan. 28, 2008 (EP) .................................. O81 010084 tives with a potential for once weekly administration. US 2011/0082079 A1 Apr. 7, 2011 GLUCAGON-LIKE PEPTDE-1 DERVATIVES 0011. A further object of the invention is to provide a long AND THER PHARMACEUTICAL USE acting, i.e. having an administration regimen as described above, GLP-1 derivative. FIELD OF THE INVENTION 0012 Another object of this invention is to provide a 0001. This invention relates to the field of therapeutic pep GLP-1 derivative with high potency (receptor affinity) in tides, i.e. to new protracted peptide derivatives of Glucagon order to reduce the therapeutic dose used for example for once Like Peptide-1 (GLP-1). weekly s.c. dosing or alternatively for non-invasive delivery. 0013 Another object of this invention is to provide a BACKGROUND OF THE INVENTION GLP-1 derivative with a high binding affinity to the extracel lular domain of the GLP-1 receptor (GLP-1R). 0002. A range of different approaches have been used for 0014) Another object of this invention is to provide a modifying the structure of GLP-1 compounds in order to GLP-1 derivative with high albumin binding affinity which provide a longer duration of action in vivo. protects the peptide for proteolytic degradation and reduce 0003 WO 2006/097535 discloses various peptide ago renal clearance of the peptide. nists of the glucagon family with secretin like activity and 00.15 Potency, stability, half-life, binding affinity to albu their therapeutic use. GLP-1 (7-37) derivatives comprising a min, and binding affinity to the extracellular domain of the modified GLP-1 (7-37) sequence are disclosed in Examples 3 GLP-1 receptor are properties of potential relevance for an and 5 thereof. These derivatives, however, do not have a Glu overall object of achieving long-acting, stable and of course residue at position 22 and an Arg residue at position 26. therapeutically active GLP-1 derivatives with a potential for 0004 WO 01/04156 discloses peptides that lower blood once weekly administration. glucose levels. Compounds 4, 5, 6, 7, 10, 11, 12, and 13 thereof are GLP-1 derivatives, however none of these com SUMMARY OF THE INVENTION pounds have a Glu residue at position 22 and an Arg residue at position 26. 0016. In one aspect of the invention, a GLP-1 derivative is 0005 EP 1364967 discloses glucagon-like insulinotropic provided which comprises a modified GLP-1 (7-37) sequence peptides, compositions and methods. None of the five GLP-1 having:ii) a total of 2-12 amino acid modifications as com compositions of Example 1 thereof comprises a modified pared to GLP-1 (7-37) (SEQ ID No: 1), including a) a Glu GLP-1 sequence having Glu at position 22 and Argat position residue at a position equivalent to position 22 of GLP-1 (7- 26. 37), and b) an Arg residue at a position equivalent to position 0006 WO 98/08871 discloses protracted GLP-1 deriva 26 of GLP-1 (7-37); and ii) which is derivatised with an albu tives including liraglutide, which is a GLP-1 derivative for min binding residue or pegylated in a position selected from once daily administration developed by Novo Nordisk A/S. a position equivalent to position 18, 20, 23, 30, 31, 34, 36,37. Liraglutide is expected to be marketed for the treatment of or 39 of GLP-1 (7-37). type 2 diabetes as of 2009. Liraglutide does not have Glu at 0017. In a further aspect, pharmaceutical compositions position 22 and Argat position 26. The derivative disclosed in and methods and uses of the derivatives according to the Example 30 of this reference does, but is not derivatised in invention, are provided. position 18, 20, 23, 30, 31, 34, 36, 37, or 39. 0007 WO 2005/027978 discloses novel GLP-1 deriva DESCRIPTION OF THE INVENTION tives including three which comprise a modified GLP-1 (7-37) sequence having a Glu at position 22 and an Arg at position Definitions and Preferred Embodiments 26. These derivatives, however, are not derivatised in position 18, 20, 23, 30, 31, 34, 36, 37, or 39. 0018. In the present specification, the following terms 0008 Runge et al (Journal of Biological Chemistry, vol. have the indicated meaning: 283, no. 17, pp. 11340-11347), which was published after the (0019. The term “polypeptide' and “peptide' as used priority dates of the present invention, discloses the crystal herein means a compound composed of at least five constitu structure of the extracellular domain of the ligand-bound ent amino acids connected by peptide bonds. The constituent GLP-1 receptor. amino acids may be from the group of the amino acids 0009. Many diabetes patients particularly in the type 2 encoded by the genetic code and they may be natural amino diabetes segment are subject to so-called “needle-phobia’, acids which are not encoded by the genetic code, as well as i.e. a Substantial fear of injecting themselves. In the type 2 synthetic amino acids. Natural amino acids which are not diabetes segment most patients are treated with oral hypogly encoded by the genetic code are e.g., Y-carboxyglutamate, cemic agents, and since GLP-1 compounds are expected to be ornithine, phosphoserine, D-alanine and D-glutamine. Syn an injectable pharmaceutical product these patients will be thetic amino acids comprise amino acids manufactured by administered, the fear of injections may become a serious chemical synthesis, i.e. D-isomers of the amino acids obstacle for the widespread use of the clinically very prom encoded by the genetic code such as D-alanine and D-leucine, ising compounds. Thus, there is a need to develop new com Aib (CL-aminoisobutyric acid), Abu (CL-aminobutyric acid), pounds which can be administered less than once daily, e.g. Tle (tert-butylglycine), B-alanine, 3-aminomethyl benzoic once every second or third day preferably once weekly, while acid, anthranilic acid. retaining an acceptable clinical profile or optionally via non 0020. The 22 proteogenic amino acids are: Alanine, Argi invasive administration Such as pulmonary, nasal, Sublingual, nine, Asparagine, Aspartic acid, Cysteine, Cystine, bucal or oral administration. Glutamine, Glutamic acid, Glycine. Histidine, Hydroxypro 0010. One object of the present invention is to provide a line, Isoleucine, Leucine, Lysine, Methionine, Phenylala chemically, physically and enzymatically stable GLP-1 nine, Proline, Serine, Threonine, Tryptophan, Tyrosine, derivative, preferably with a high alpha-helical content. Valine. US 2011/0082079 A1 Apr. 7, 2011 0021. Thus a non-proteogenic amino acid (which may also 37) sequence (having the sequence of SEQID No: 1). Corre be designated a non-natural amino acid) is a moiety which can sponding positions are easily deduced, e.g. by simple hand be incorporated into a peptide via peptide bonds but is not a writing and eyeballing. In the alternative, a standard protein proteogenic amino acid. Examples are y-carboxyglutamate, or peptide alignment program may be used, such as “align” ornithine, phosphoserine, the D-amino acids such as D-ala which is a Needleman-Wunsch alignment. The algorithm is nine and D-glutamine, Synthetic non-proteogenic amino described in Needleman, S. B. and Wunsch, C. D., (1970), acids comprising amino acids manufactured by chemical Syn Journal of Molecular Biology, 48: 443-453, and the align thesis, i.e. D- isomers of the amino acids encoded by the program by Myers and W. Miller in "Optimal Alignments in genetic code such as D-alanine and D-leucine, Aib C-ami Linear Space’ CABIOS (computer applications in the bio noisobutyric acid), Abu (CL-aminobutyric acid), Tle (tert-bu sciences) (1988) 4:11-17. For the alignment, the default scor tylglycine), 3-aminomethyl benzoic acid, anthranilic acid, ing matrix BLOSUM50 and the default identity matrix may des-amino-histidine (abbreviated DesaminoHis, alternative be used, and the penalty for the first residue in a gap may be name imidazopropionic acid, abbreviated Impr), the betaana set at -12 and the penalties for additional residues in a gap at logs of amino acids such as B-alanine etc., D-histidine, -2.