Exendin-4 derivatives

GLP1fGIP or trigonal dual agonists of GLP1fGIPfglucagon

FIELD OF THE INVENTION

The present invention relates to exendin-4 peptide analogs that activate the glucagon-like peptide 1 receptor (GLP-1) and glucose dependent insulinotropic polypeptide (GIP) and optionally the glucagon receptor (GCG) and its medical use , for example, in the treatment of metabolic syndrome disorders, which include diabetes and obesity, in addition to the reduction of excessive food intake.

BACKGROUND OF THE INVENTION

Exendin-4 is a peptide of 39 amino acids that is produced by the salt glands of the Gila monster (Heloderma suspectum) (Eng. J. et al., J. Biol. Chem., 267: 7402-05, 1992). . Exendin-4 is a glucagon-like peptide-1 receptor (GLP-1) receptor activator, considering that it shows only very low GIP receptor activation and does not activate the glucagon receptor (see Table 1).

Table 1. Powers of exendin-4 in human GLP-1, GIP and glucagon receptors (indicated in pM) at increasing concentrations and measuring cAMP formed as described in Methods

<dl><dt>SE ID NO: </dt><dd>peplified EC50 of hGLP-1 R [pM] CESG of hGIP R [pM] Cessation of hGlucagon R [pM] </dd></dl>

<dl><dt>1 </dt><dd>exendin-4 0.4 12500.0 &gt; 10000000 </dd></dl>

Exendin-4 shares many of the glucorregulatory actions observed with GLP-1 Clinical and non-clinical studies have shown that exendin-4 has several beneficial antidiabetic properties that include a glucose-dependent enhancement in insulin synthesis and secretion, dependent suppression of glucose from glucagon secretion, slowing of gastric emptying, reduction of food intake and body weight, and an increase in beta cell mass and markers of beta cell function (Gentilella R et al., Diabetes Obes Metab., 11: 544-56, 2009; Norris SL et al., Oiabet Med., 26: 837-46, 2009; Bunck MC et al., Diabetes Care .. 34: 2041-7, 2011)

These effects are beneficial not only for diabetics, but also for patients suffering from obesity. Patients with obesity have a higher risk of diabetes, hypertension, hyperlipidemia, cardiovascular and musculoskeletal diseases.

With respect to GLP-1 and GIP, exendin-4 is more resistant to cleavage by dipeptidyl peptidase-4 (DPP4), producing a prolonged in vivo half-life and duration of action (Eng J., Diabetes, 45 (SuppI 2 ): 152A (abstract 554), 1996; Deacon CF, Horm Melab Res, 36: 761-5, 2004).

It was also shown that exendin-4 was much more stable towards degradation by neutral endopeptidase (NEP), when compared with GLP-1, glucagon or oxintomodulin (Druce MR et al., Endocrinology, 150 (4), 1712-1721 , 2009)

However, exendin-4 is chemically labile due to the oxidation of methionine at position 14 (Hargrove DM et al., Regul. Pepl., 141. 113-9, 2007), in addition to deamidation and isomerization of the asparagine in position 28 (WO 2004/035623).

The amino acid sequence of exendin-4 is shown as SE ID NO:

HGEGTFTSoLSKoMEEEAVRLFIEWLKNGGPSSGAPPPS-NH2

The amino acid sequence of GLP-1 (7-36} -amide is shown as SEa ID NO · 2

HAEGTFTSDVSSYLEGaAAKEFIAWLVKGR-NH2

Liraglutide is a marketed chemically modified GLP-1 analogue in which, among other modifications, a fatty acid is linked to a lysine in position 20 leading to a prolonged duration of action (Drucker DJ et al., Nature Drug Disc Rev. 9, 267-268, 2010; Buse, JB et al., Lancet, 374: 39-47, 2009)

The amino acid sequence of liraglutide is shown as SEO ID NO: 3.

HAEGTFTSDVSSYLEGQAAK ((S) -4-Carboxy-4-hexadecanoylamino-butyryl-)

EFIAWLVRGRG-OH

GIP (glucose dependent insulinotropic polypeptide) is a 42 amino acid peptide that is released from intestinal K cells after food consumption. GIP and GLP-1 are the two hormones derived from enteroendocrine cells of the intestine that represent the incretin effect, which represents more than 70% of the insulin response to oral glucose exposure (Baggio LL, Drucker DJ, Biology of incretins · GLP-1 and GIP Gastroenterology 2007; 132: 2131-2157).

The amino acid sequence of GIP is shown as SEQ ID NO: 4 ·

YAEGTFISDYSIAMDKIHQQDFVNWLLAQKGKKNDWKHNITQ-OH

Glucagon is a peptide of 29 amino acids that is released into the bloodstream when glucose in circulation is low. The amino acid sequence of glucagon is shown in SEQ ID NO: 5

HSQGTFTSDYSKYLDSRRAQDFVQWLMNT-OH

During hypoglycemia, when blood glucose levels drop below normal, glucagon sends a signal to the liver to degrade glycogen and release glucose, causing an increase in blood glucose levels to reach a normal level. Hypoglycaemia is a common side effect of patients treated with insulin with hyperglycaemia (elevated blood glucose levels) due to diabetes. Thus, the most predominant function of glucagon in the regulation of glucose is to counteract the action of insulin and maintain blood glucose levels.

Holst (Holst, JJ Physiol, Rev 2007,87, 1409) and Meier (Meier, JJ Nat. Rev. Endocrinol, 2012, 8, 728) describe that GLP-1 receptor agonists, such as GLP-1, liraglutide and exendin-4, improve glycemic control in patients with T2DM, reducing fasting and postprandial glucose (FPG and PPG). Peptides that bind to and activate the GLP-1 receptor are described in patent applications WO 98108871 A1, W02008 / 081418 A1 and W02008 / 023050 A1.

WO 2008/081418 A1 discloses glucagon-like protein 1 receptor-like target compounds, comprising GLP-1 receptor targeting conjugated agent-conjugate conjugates to an antibody combining site, in addition to uses of such compounds that include diabetes prevention or treatment methods

or conditions related to diabetes

WO 2008/023050 A1 refers to therapeutic peptides derived from exendin-4, in which a lysine is derivatized to give an acylated lysine residue

It has been described that dual activation of GLP-1 and GIP receptors, for example, by combining the actions of GLP-1 and GIP in a preparation, leads to a therapeutic principle with significantly better reduction of blood glucose levels , high insulin secretion and reduced body weight in mice with T2DM and obesity in comparison with the commercialized GLP-1 agonist liraglutide (for example, VA Gault et al., Clin Sci (Lond), 121, 107-117, 2011) . It was demonstrated that native GLP-1 and GIP in humans after ca-infusion interacted in an additive manner with a significantly elevated insulinotropic effect compared to GLP-1 alone (MA Nauck et al., J. Clin. Endocrinol. Metab. , 76, 912-917, 1993)

The design of hybrid molecules that combine agonism on the GLP-1 receptor, the GIP receptor and glucagon offers the therapeutic potential to achieve a significantly better reduction of blood glucose levels, high insulin secretion and a significant effect even more pronounced on the reduction of body weight compared to the commercialized GLP-1 agonist liraglutide (eg VA Gault et al., Clin Sci (Lond), 121, 107-117, 2011).

The compounds of the present invention are derivatives of exendin-4, which show agonist activity at the GLP-1 and GIP receptor and optionally the glucagon receptor and which have -among others- preferably the following modifications: Tyr at position 1 e lIe in position 12

Surprisingly, it was found that modification of the selective GLP-1R agonist exendin-4 by Tyr at position 1 and lIe at position 12 produces a peptide with high dual activity at the GLP-1 and GIP receptors. This observation is surprising , since the same modification in other GLP-1 agonists, such as GLP-1 itself, does not produce high activity in the GIP receptor, as shown in Table 2.

Table 2 · Powers of peptide analogues exendin-4 and GLP-1 in GLP-1 and GIP receptors (indicated in pM) at increasing concentrations and measuring cAMP formed as described in Methods.

<dl><dt>SEQ ID NO · </dt><dd>peptide CE50 of hGIP R [pM] EC50 of hGLP 1 R [pM] </dd></dl>

<dl><dt>6 </dt><dd>Tyr (1) lIe (12) -exendin-4 93, 9 1,3 </dd></dl>

<dl><dt>7 </dt><dd>Tyr (1) lIe (12) GLP1 3660.0 5.0 </dd></dl>

Peptides that bind and activate both the GIP receptor and the GLP-1 receptor and optionally the glucagon receptor, and improve glycemic control, suppress body weight gain and reduce food consumption are described in the patent WO 2011/119657 A1, WO 2012/138941 A1, WO 2010/011439 A2, WO 2010f148089 A1, WO 2011 f094337 A1, WO 2012f088116 A2_ These applications disclose that mixed agonists of the GLP-1 receptor, the GIP receptor and optionally the glucagon receptor as analogs of the native GIP or glucagon sequences.

The compounds of the present invention are analogs of the exendin-4 peptide comprising leucine in the 10-position and glutamine in the 13-Krstenansky et al. (Biochemistry, 25, 3833-3839, 1986) show the importance of residues 10 to 13 of glucagon for their interactions with adenylate cyclase receptor and activation. In the exendin-4 peptide analogs of the present invention, several of the underlying residues are different from said glucagon. In particular, the Tyr10 and Tyr13 residues are substituted with leucine at the 10-position and glutamine, a non-aromatic polar amino acid, at the position 13_ This substitution, especially in combination with isoleucine at position 23 and glutamate at position 24,

In addition, the compounds of the present invention are derivatives of exendin-4 with fatty acid acyl residues in the 14-position. This functionalization of the fatty acid in the 14-position produces an improved pharmacokinetic profile. Surprisingly, the functionalization of the fatty acid in the 14-position also leads to peptides with a significantly higher GIPR activity, for example those shown in Example 5, Table 8.

The compounds of the present invention are analogs of the exendin-4 peptide containing alpha, alpha dialkylated amino acids with a basic side chain at the 20-position. Surprisingly, modifying the sequence of the exendin-4 with one of these amino acids leads to compounds with an improved biophysical profile, such as the solubility behavior (particularly at low pH, especially at pH 4.5) or aggregation in solution, when the non-natural amino acid is incorporated in position 20_ The resultant exendin-4 analogues thus maintain their high activity in the GLP-1 receptor, the GIP receptor and optionally the glucagon receptor. The incorporation of these non-natural amino acids also increases the enzymatic stability of the peptides, possibly producing improved pharmacokinetic properties.

BRIEF SUMMARY OF THE INVENTION

Exendin-4 analogs that potently activate the GLP-1 and GIP receptor and optionally the glucagon receptor are provided herein. In these exendin-4 analogs, among other substitutions, methionine at position 14 is substituted with an amino acid bearing a -NH1 group on the side chain, which is further substituted with a lipophilic side chain (for example, a fatty acid optionally combined with a linker)

The invention provides a peplidic compound having the formula (1):

R1 _Z _R2 (1)

wherein Z is a peptide residue having the formula (11)

Tyr-Aib-X3-Gly-Thr -Phe-Thr -Ser-Asp-Leu-Ser -X 12 -Gln-X 14-X 15-X 16-X 1 7 -X 18-X 19 -X20-X21- PheIle-Glu-T rp-Leu-L ys-X28-X29-Gly-Pro-Ser -Ser -Gly-Ala-Pro-Pro-Pro-Ser -X40 (11)

X3 represents an amino acid residue selected from Gln, Glu and His,

X12 represents an amino acid residue selected from lIe and Lys,

X14 represents an amino acid residue that has a side chain with an -NH2 group, in which the side chain group -NH2 is functionalized by -C (0) -R5, in which RS can be a residue comprising up to 50

or up to 100 carbon atoms and optionally heteroatoms selected from halogen, N, O, S and / or P,

X15 represents an amino acid residue selected from Asp and Glu,

X16 represents an amino acid residue selected from Ser, Lys, Glu and Gln,

X17 represents an amino acid residue selected from Arg, Lys, Glu, Gln, Leu, Aib, Tyr and Ala,

X18 represents an amino acid residue selected from Ala, Arg, Aib, Leu and Tyr,

X19 represents an amino acid residue selected from Ala, Val and Aib,

X20 represents an amino acid residue selected from Pip, (S) MeLys, (R) MeLys and (S) MeOrn, X21 represents an amino acid residue selected from Asp, Glu and Leu,

X28 represents an amino acid residue selected from Asn, Ala, Aib and Ser,

X29 represents an amino acid residue selected from Gly, Thr, Aib, D-Ala and Ala,

X40 is absent or represents Lys,

R 'represents NH2,

R2 represents OH or NH2.

or a solvate salo thereof.

The compounds of the invention are GLP-1 and GIP receptor agonists and optionally glucagon receptor agonists as determined by the observation that they can stimulate the formation of intracellular cAMP. The determination of in vitro potency in cellular agonist assays is quantified by determining the concentrations that produce 50% activation of the maximum response (CESO) as described in Methods.

In certain embodiments, the invention, therefore, provides a peptide compound having the formula (1):

(1) wherein Z is a peptide residue having the formula (11) Tyr-Aib-X3-Gly-Thr -Phe-Thr -Ser -Asp-Leu-Ser -X 12 -Gln-X 14-X 1S-X 16-X 1 7 -X 18-X 19-X20-X21-Phe

Ile-Glu-T rp-Leu-L ys-X28-X29-Gly-Pro-Ser -Ser -Gly-Ala-Pro-Pro-Pro-Ser -X40 (11) X3 represents an amino acid residue selected from Gln, Glu and His, X12 represents an amino acid residue selected from lIe and Lys, X14 represents an amino acid residue having a side chain with an -NH2 group, in which the group of

laleral chain -NH2 is functionally raised by _C (0) _R5, wherein R5 is a moiety comprising up to 50 or up to 100 carbon atoms and optionally heteroatoms selected from halogen, N, O, S and P, X15 represents an amino acid residue selected from Asp and Glu, X16 represents an amino acid residue selected from Ser, Lys, Glu and Gln, X17 represents an amino acid residue selected from Arg, Lys, Glu, Gln, Leu, Aib, Tyr and Ala, X18 represents an amino acid residue selected from Ala, Arg, Aib, Leu and Tyr, X19 represents an amino acid residue selected from Ala, Val and Aib, X20 represents an amino acid residue selected from Pip, (S) MeLys, (R) MeLys and (S) MeOm, X21 represents an amino acid residue selected from Asp, Glu and Leu, X28 represents an amino acid residue selected from Asn, Ala, Aib and Ser, X29 represents an amino acid residue selected from Gly, Thr, Aib, D-Ala and Ala, X40 is absent or represents Lys, R 'represents NH2, R2 represents OH or NH2

or a solvate salt thereof, wherein the peptide compound has a relative activity of at least 0.04%, preferably at least 0.08%, more preferably at least 0.2% as compared to GIP natural in the GIP receiver.

In addition, the peptide compound, particularly with a lysine at position 14 which is further substituted with a lipophilic moiety, has a relative activity of at least 0.07%, preferably at least 0.1%, more preferably at least 0.14%, more preferably at least 0.35% and even more preferably at least 0.4% compared to that of GLP-1 (7-36) at the GLP-1 receptor. In addition, the peptide compound , particularly with a lysine at position 14 which is further substituted with a lipophilic moiety, exhibits a relative activity of at least 0.04% (ie, CEsolt; 1000 pM), more preferably 0.08% (i.e. , CEsolt; 500 pM) and even more preferably 0.2% (ie, CEsolt; 200 pM) compared to that of natural GIP in the GIP receptor (EC so'quot; 0.4 pM)

Optionally, in some embodiments, the peptide compound, particularly with a lysine at position 14 which is further substituted with a lipophilic moiety, exhibits a relative activity of at least 0.1%, preferably at least 0.2%, more preferably at least 0.3%, more preferably at least 0.4% and even more preferably at least 0.5% compared to that of the natural glucagon at the glucagon receptor.

The term "activity" as used herein, it preferably refers to the ability of a compound to activate the human GLP-1 receptor, the human GIP receptor and optionally the human glucagon receptor. More preferably, the term "activity" as used herein refers to the ability of a compound to stimulate the formation of intracellular cAMP. The term "relative activity" as used herein it is understood that it refers to the ability of a compound to activate a receptor in a certain relationship with respect to another receptor agonist or with respect to another receptor. The activation of the receptors by the agonists (for example, by measuring the level of cAMP) is determined as described herein, for example,

According to one embodiment, the compounds of the invention have an EC so for the hGLP-1 receptor of 500 pM or less, preferably 200 pM or less; more preferably 150 pM or less, more preferably 100 pM or less, more preferably 90 pM or less, more preferably 80 pM or less, more preferably 70 pM or less, more preferably 60 pM or less, more preferably of 50 pM or less. more preferably 40 pM or less, more preferably 30 pM or less, and more preferably 20 pM or less

According to one embodiment, the compounds of the invention have a CE for the hGIP receptor of 500 pM or less. preferably 200 pM or less; more preferably 150 pM or less, more preferably 100 pM or less, more preferably 90 pM or less, more preferably 80 pM or less, more preferably 70 pM or less, more preferably 60 pM or less, more preferably 50 pM or less, more preferably 40 pM or less, more preferably 30 pM or less, and more preferably 20 pM or less

According to another embodiment, the compounds of the invention optionally have a HGlucagon receptor CE of 500 pM or less, preferably 200 pM or less; more preferably 150 pM or less, more preferably 100 pM or less, more preferably 90 pM or less, more preferably 80 pM or less, more preferably 70 pM or less, more preferably 60 pM or less, more preferably 50 pM or less, more preferably 40 pM or less, more preferably 30 pM or less, and more preferably 20 pM or less

According to another embodiment, the compounds of the invention have a CE for the hGLP-1 receptor of 500 pM or less. preferably 200 pM or less; more preferably 150 pM or less, more preferably 100 pM or less, more preferably 90 pM or less, more preferably 80 pM or less, more preferably 70 pM or less, more preferably 60 pM or less, more preferably of 50 pM or less, more preferably of 40 pM or less, more preferably of 30 pM or less, and more preferably of 20 pM or less, and / or an EC so for the hGIP receptor of 500 pM or less, preferably of 200 pM or less; more preferably 150 pM or less, more preferably 100 pM or less, more preferably 90 pM or less, more preferably 80 pM or less, more preferably 70 pM or less, more preferably 60 pM or less, more preferably 50 pM or less, more preferably 40 pM or less. more preferably 30 pM or less, and more preferably 20 pM or less, and / or optionally a CE for the hGlucagon receptor of 500 pM or less, preferably 200 pM or less; more preferably 150 pM or less, more preferably 100 pM or less, more preferably 90 pM or less. more preferably 80 pM or less, more preferably 70 pM or less, more preferably 60 pM or less, more preferably 50 pM or less, more preferably 40 pM or less, more preferably 30 pM or less, and more preferably 20 pM or less and / or optionally a CE for the hGlucagon receptor of 500 pM or less, preferably 200 pM or less; more preferably 150 pM or less, more preferably 100 pM or less, more preferably 90 pM or less. more preferably 80 pM or less, more preferably 70 pM or less, more preferably 60 pM or less, more preferably 50 pM or less, more preferably 40 pM or less, more preferably 30 pM or less, and more preferably 20 pM or less and / or optionally a CE for the hGlucagon receptor of 500 pM or less, preferably 200 pM or less; more preferably 150 pM or less, more preferably 100 pM or less, more preferably 90 pM or less. more preferably 80 pM or less, more preferably 70 pM or less, more preferably 60 pM or less, more preferably 50 pM or less, more preferably 40 pM or less, more preferably 30 pM or less, and more preferably 20 pM or less

In another further embodiment, the CEso for both receptors, ie, for the hGLP-1 receptor and for the hGIP receptor, is 500 pM or less, more preferably 200 pM or less, more preferably 150 pM or less, more preferably 100 pM or less, more preferably 90 pM or less, more preferably 80 pM or less, more preferably 70 pM or less, more preferably 60 pM or less, more preferably 50 pM or less, more preferably 40 pM or less, more preferably 30 pM or less, more preferably pM or less, more preferably 20 pM or less

In another further embodiment, the CE for the three receptors, ie, for the hGLP-1 receptor, for the hGIP receptor and for the hGlucagon receptor, is 500 pM or less, more preferably 200 pM or less, more preferably 150 pM or less, more preferably 100 pM or less, more preferably 90 pM or less, more preferably 80 pM or less, more preferably 70 pM or less, more preferably 60 pM or less, more preferably 50 pM or less, more preferably 40 pM or less, more preferably pM or less, more preferably 30 pM or less, more preferably 20 pM or less.

The CE for the hGLP · 1 receptor, hGIP receptor and hGlucagon receptor can be determined as described in Methods herein and as used to generate the results described in Example

5.

The compounds of the invention have the ability to reduce intestinal transit, increase gastric content and / or reduce a patient's food intake. These activities of the compounds of the invention can be evaluated in animal models known to the skilled artisan and also described herein in Methods. The results of such experiments are described in Example 10. Preferred compounds of the invention can increase the gastric content of mice, preferably of female NMRI mice, if administered as a single dose, preferably subcutaneously, of 0.02 mg / ml. kg of body weight to at least 25%, more preferably to at least 30%, more preferably to at least 40%, more preferably to at least 50%, more preferably to at least 60%,

Preferably, this result is measured 1 h after the administration of the respective compound and 30 min after the administration of a bolus, and / or reduces the intestinal transit of mice, preferably of female NMRI mice, if administered as a single dose, preferably subcutaneous dose, from 0.02 mgfkg of body weight to at least 45%; more preferably at least 50%, more preferably at least 55%, more preferably at least 60%, and more preferably at least 65%; and / or reduce the food intake of mice, preferably female NMRI mice, over a period of 22 h, if administered as a single dose, preferably subcutaneously, from 0.01 mg / kg of body weight to at least 10 %, more preferably 15%, and more preferably 20%

The compounds of the invention have the ability to reduce the blood glucose level and / or reduce HbA1c levels of a patient. These activities of the compounds of the invention can be evaluated in animal models known to the skilled person and are also described herein in Methods. The results of such experiments are described in Examples 8 and 9.

Preferred compounds of the invention can reduce the blood glucose level of mice, preferably female diabetic db / db female mice deficient in the leptin receptor, for a period of 24 h, if administered as a single dose, preferably subcutaneous dose , from 0.01 mgfkg of body weight to at least 4 mmolfl; more preferably at least 6 mmolll, more preferably at least 8 mmolfl. If the dose is increased to 0.1 mg / kg of body weight, a more pronounced reduction in blood glucose levels in mice can be observed over a period of 24 h, if administered as a single dose, preferably subcutaneously. Preferably, the compounds of the invention lead to a reduction of at least 7 mmolll; more preferably at least 9 mmolll, more preferably at least 11 mmolll.

The compounds of the invention also have the ability to reduce a patient's body weight. These activities of the compounds of the invention can be evaluated in animal models known to the skilled person and are also described herein in Methods and in Example 7.

Surprisingly, it was found that the peptide compounds of formula (1), particularly those with a lysine (or similar analogues) at position 14 which is further substituted with a lipophilic moiety, showed activation of very potent GLp · 1 and GIP receptors. ; additionally in combination with amino acids such as Gln in position 3, very potent glucagon receptor activation can also be provided

It is described in the literature (Murage EN et al., 8ioorg, Med. Chem. 16 (2008), 10106 · 10112), that a GLP-1 analog with an acetylated lysine in pos. 14 showed significantly reduced potency compared to natural GLP-1.

In addition, the oxidation (in vitro or in vivo) of methionine, present in the central structure of exendin-4, is no longer possible for the peptide compounds of formula (1)

In addition, the compounds of the invention preferably have high solubility at acidic and / or physiological pH values, for example, at pH 4.5 and pH 7.4 at 25 OC, in another embodiment at least 0.5 mg / ml and in a particular embodiment at least 1.0 mg / ml.

In addition, according to one embodiment, the compounds of the invention preferably have high stability when stored in solution. The preferred test conditions for determining stability is storage for 7 days at 25 ° C in solution at pH 4.5 or pH 7.4. The remaining amount of peptide is determined by chromatographic analyzes as described in Methods and Examples. Preferably, after 7 days at 25 ° C in solution at pH 4.5 or pH 7.4, the remaining amount of peptide is at least 80%, more preferably at least 85%, even more preferably at least 90% and even more preferably at least 95%

Preferably, the compounds of the present invention comprise a Z peptide moiety (formula 11) which is a linear sequence of 39-40 aminocarboxylic acids, particularly a-aminocarboxylic acids linked by peptide, ie, carboxamide bonds.

In one embodiment, the X14 position represents an amino acid residue with a side chain group -NH2 functionalized, such as Lys, Orn, Dab or functionalized Dap, more preferably functionalized Lys and X40 is absent or represents Lys

An amino acid residue with a side chain group -NH2, for example Lys, Orn, Dab or DaR, is functionalized by the fact that at least one H atom of the side chain group -NH2 is substituted by _C (O) -Rs , wherein R5 is a moiety comprising up to 50 or up to 100 carbon atoms and optionally heteroatoms selected from halogen, N, O, S and / or P

In certain embodiments, RS may comprise a lipophilic moiety, for example a straight or branched acyclic saturated hydrocarbon group, wherein R 5 particularly comprises a saturated or unsaturated saturated (C 4 -C 30) acyclic hydrocarbon group, linear or branched, and a cyclic group saturated, unsaturated or aromatic, particularly a mono-, bi- or tricyclic group comprising 4 to 14 carbon atoms and 0.1 I heteroatoms selected from N, OYS, for example cyclohexyl, phenyl, biphenyl, chromanyl, phenanthrenyl or naphlyl, wherein the acyclic or cyclic group can be unsubstituted or substituted, for example, by halogen, -OH and C02H.

More preferred R5 groups may comprise a lipophilic moiety, for example a straight or branched acyclic, saturated or unsaturated hydrocarbon group (C 12 "quot; Cn). The lipophilic moiety can be attached to the side chain group -NH2 by a linker in all stereoisomeric pons, for example a linker comprising one or more, for example 2, 3 or 4 amino acid linking groups such as y-aminobutyric acid (GASA ), t-aminohexanoic acid (t-Ahx), y-Glu yfo ~ -Ala. In one embodiment, the lipophilic moiety is linked to the side chain -NH2 group by a linker. In another embodiment, the lipophilic moiety is directly linked to the side chain group -NH2. Specific examples of amino acid linking groups are (~ -Ala) 1-4, (y-GIU) 1-4, (E-Ahx) 1- 4 o (GASAh.4) Preferred amino acid connecting groups are f3, -Ala, y-Glu, f3, -A1a-f3,

Specific preferred examples of groups C (O) -R5 are listed in the following Table 3, which are selected from the group consisting of (S) -4-carboxy-4-hexadecanoylamino-butyryl-, (8} -4-carboxy- 4-Qctadecanoylamino-butyryl-, 4-hexadecanoylamino-buli ril-, 4- {3 - [(R} -2.5, 7, 8-telramelyl-2 - ((4R, SR) -4, S, 12 -trimelil -Iridecil) -chroman -6 · iloxic rbonyl) propionylamino} -butyryl, 4-octadecanoylamine-butyryl-, 4- ((Z) -octadec-9-enylamino) -butyryl-, 6 - [(4, 4 -diphenyl-cyclohexyloxy) hydroxy-phosphoryloxy] -hexanoyl, hexadecanoyl-, {S) -4-carboxy-4- (lS-carboxy-pentadecanoylamino) -butyryl-, (S) -4-carboxy-4 {3- [3 - {(2S, 3R, 4S, SR) -S-carboxy-2,3,4, S-tetrahydroxy-pentanoylamino) -propionylamino] -propionylamino} -butyryl-, (S) -4carboxy-4- {3 - [{R) -2, 5, 7, a-tetra me! Il-2 - {{4 R, aR) -4, 8, 12-! Rim e!il-tridecyl) -chroman-6-yloxycarbonyl] -propionylamino} -butyryl-, (S) -4-ca rboxi-4 - {{9Z, 12Z) -octadeca-9, 12 -dienoylamino) -butyryl- , (S) -4-ca rboxi-4- [6. {{2S, 3R, 4S, SR) -S-ca rboxi-2,3, 4, 5-hydrohydroxy-penlanoylamino) -hexanoylamino] -bubiryl-, (S) ) -4-carboxy-4- «2S, 3R, 4S, 5R} -5-carboxy-2,3,4,5-lelrahydroxypenlanQinylamino) -bubiryl-, (S) -4-carboxy-4-lelradeca noilam -bubiryl-, (S} -4- (l 1 -benzyloxy rbon il-undecanylamino) -4carboxy -bubiryl-, (S} -4-carboxy-4- [11- ((28, 3R, 4R, SR) -2, 3.4, S, 6-penlah idroxy-hexylcarbamoyl) -undecanoylamine] -bubyryl-, (S) -4-ca rboxi-4- ((Z} -ocladec-9-enoylamino) -butyryl-, (S) -4-ca-rboxi-4- (4-dodecyloxy-benzoylamino) -butyryl, (S) -4-carboxy-4-henicosanoylamino-butyryl-, (S) -4-carboxy-4-docosanoylamino-butyryl-, (S} -4-carboxy-4- ((Z) -nonadec-l O-enoylamino) buliryl-,(S) -4-carboxy-4- (4-decyloxy-benzoylamino) -butyryl., (S) -4-carboxy-4 - [(4'-cyloxy-biphenyl-4-carbonyl) -a mino] - buliril-, (S) -4-ca rboxi-4- (12-phenyl-dodecanylamino) -butyryl-, (S} -4-carboxy-4-icosa noilam ino-buliryl-, (S) -4- carboxy-4- ((S} -4-carboxy-4-hexadecanoylamino-butyrylamino} -butyryl-, (S) -4-carboxy-4- 'S) -4-carboxy-4-octadecanoylamino-butyrylamino) bUyryl-, 3- (3-ocladecanoylamino-propionylamino) -propionyl-, 3- (3-hexadecanoylamino-propionylamino) -propionyl-, 3-hexadecanoylamino-propionyl-, (S) -4-carboxy-4 - [{R) -4 - {{3R, SS, 7R, aR, 9R, I OS, 12S, 13R, 14R, 17R) -3,7, 12-trihydroxy, l, 13-lrim ethyl-hexadecah idro-cyclopenla [a] phena nlhren-17 -il) pentylamino) -bubiryl-, (S) -4-carboxy-4- {(R) -4 "3R, 5R, SR, 9S, 1 OS, 13R, 14S, 17R) -3-hydroxy-1O, 13-dim elil-hexadecah idro-cyclopenla [a] phenanlren -17-yl) pentanoylamino] -bubiryl-,(S) -4-carboxy-4 - {{9S, 10R) -9,1-O, 16-trihydroxy-hexadecanoylamino) -butyryl-, tetradecanoyl-, 11-carboxy-undecanoyl-, l-benzyloxy-rbonyl-undecanoyl-, (S ) -4-ca rboxi-4- ((S} -4-carboxy-4-lelradeca-noylaminobutyrylamino} -bubyryl-, 6- [hydroxy- (naphthalene-2-yloxy) -phosphoryloxy] -hexanoyl-, 6- {hydroxy} - (5-phenyl-pentyloxy} -phosphoryloxy) hexanoyl-, 4- (Naphthalene-2-sulfonylamino) -4-oxo-butyryl-, 4- (biphenyl-4-sulfonyl-amino) -4-oxo-bulyryl., ( S) -4-carboxy-4 - {(S} -4ca rboxi -4- {2 - (2 - {2- {2- (2- {2 - [(S) -4-ca rboxi-4- ( 17 -carboxy-he pladeca noilamino) -bubyrylamino] -eloxy} -eloxy) -acetyl amino) ethoxy} -ethoxy) -acetylamino] -butyrylamino} -bubyryl-, (S) -4-carboxy-4. [2- (2 - {2 - [2 - (2 - {2 - [(S) -4-carboxy-4- (1 7 -carboxyheptadecanoylamino) -butyrylamino) -ethoxy} -ethoxy) -acetylamino] -ethoxy} -ethoxy) -acetylamino) -butyryl-,(S) -4-carboxy-2 - {(S) -4ca rboxi -2 - {2 - (2 - {2- {2- (2- {2 - [(S) -4-ca rboxi-4- (17 -carboxy-he pladeca nolamino Q) -bubyrylamino] -eloxy} -eloxy) -acetyl amino) ethoxy} -ethoxy) -acetylamino] -butyrylamino} -bubiryl-, (S) -4-carboxy-2 - [2 - (2 - {2 - [2 - (2 - {2 - [(S) -4-carboxy-4- (1 7 -carboxyheptadecanoylamino) -butyrylamino] -ethoxy-eloxy) -acetylamino] -ethoxy} -ethoxy) -acetylamino] -butyryl-, (S) -4-carboxy-4 - {(S} -4ca rboxi -4- [2 - (2 - {2 - [(S} -4 -carboxy-4- (17- rboxi-heptadeca noylamino) -butyrylamino) -ethoxy} -ethoxy) -acetylamino) bulyrylamino} -bubiryl-, (S) -4-carboxy-4- [2 - (2 - {2 - [(S) -4- carboxy-4- (17 -carboxy-hepladecanoylamino) -bubyrylamino] -eloxy} ethoxy) -acetylamino] -butyryl-, (S) -4-carboxy-2 - {(S} -4-ca(S) -4-carboxy-2 - [2 - (2 - {2 - [2 - (2 - {2 - [(S) -4-carboxy-4- (1 7 -carboxyheptadecanoylamino) -butyrylamino] -ethoxy -eloxi) -acetylamino] -ethoxy} -ethoxy) -acetylamino] -butyryl-, (S) -4-carboxy-4 - {(S} -4ca rboxi -4- [2 - (2 - {2 - [( S} -4-carboxy-4- (17-ca rboxi-heptadeca noylamino) -butyrylamino) -ethoxy} -ethoxy) -acetylamino) bulyrylamino} -bubiryl-, (S) -4-carboxy-4- [2 - ( 2 - {2 - [(S) -4-carboxy-4- (17 -carboxy-hepladecanoylamino) -bubyrylamino] -eloxy} ethoxy) -acetylamino] -butyryl-, (S) -4-carboxy-2 - { (S} -4-ca(S) -4-carboxy-2 - [2 - (2 - {2 - [2 - (2 - {2 - [(S) -4-carboxy-4- (1 7 -carboxyheptadecanoylamino) -butyrylamino] -ethoxy -eloxi) -acetylamino] -ethoxy} -ethoxy) -acetylamino] -butyryl-, (S) -4-carboxy-4 - {(S} -4ca rboxi -4- [2 - (2 - {2 - [( S} -4-carboxy-4- (17-ca rboxi-heptadeca noylamino) -butyrylamino) -ethoxy} -ethoxy) -acetylamino) bulyrylamino} -bubiryl-, (S) -4-carboxy-4- [2 - ( 2 - {2 - [(S) -4-carboxy-4- (17 -carboxy-hepladecanoylamino) -bubyrylamino] -eloxy} ethoxy) -acetylamino] -butyryl-, (S) -4-carboxy-2 - { (S} -4-ca(S) -4-carboxy-4- [2 - (2 - {2 - [(S) -4-carboxy-4- (17 -carboxy-hepladecanoylamino) -bubyrylamino] -eloxy} ethoxy) -acetylamino] - butyryl-, (S) -4-carboxy-2 - {(S} -4-ca(S) -4-carboxy-4- [2 - (2 - {2 - [(S) -4-carboxy-4- (17 -carboxy-hepladecanoylamino) -bubyrylamino] -eloxy} ethoxy) -acetylamino] - butyryl-, (S) -4-carboxy-2 - {(S} -4-carboxi-2 - [2- (2 - {2 - [(S) -4-ca rboxi-4- (17-carboxy-heptadecanylamino) bulylamino) -eloxi} -eloxy) -acetylamino] -bubirila mino} -bubiril -, (S) -4-ca rboxi-2- [2- (2- {2- [(S) -4-carboxy-4- (1 7 -ca rboxihepladeca noilamno) -bubirilamno) -eloxi} - eloxi) -acetyla mino] -butyryl-, 2- (2 - {2 - [2- (2- {2- {(S) -4-ca rboxi-4- (1 7 -carboxyhepta-decanoylamino) -b ulyrylamino ] -ethoxy} -ethoxy} -acetylamino) -ethoxy} -ethoxy) -acetyl-, 2- (2 - {2 - [(S) -4-carboxy-4- (1 7 -carboxyhepdecanelamino) -bubyrylamino) -oxoxy } -eloxi) -acetyl, (S) -4-carboxy-4- «S} -4-carboxy-4 - {(S) -4-carhoxy-4 - [(S) -4carboxy-4- (19- carboxy-nonadecanoylamino) -butyrylamino] -butyryl, mino} -buliri lamino) -butyryl, 2- (2 - {2 - [2- (2 - {2 - [(S} -4carboxi -4- (16-1 H- telrazol-5-yl-hexadecan-ola-mino) -bubiryl-mino) -eloxy} -eloxy) -acetylamino] -ethoxy} -eloxy) -acetyl +,2- (2- (2 [2- (2- {2 - [(S) -4-carboxy-4- (16-carboxy-hexadecanoylamino) -bubyrylamino] -ethoxy) -ethoxy) -acetylamino] -ethoxy} - ethoxy) -acetyl-, {S) -4-carboxy-4 - {(S) -4-carboxy-4- [(8) -4-cartloxy-4- (17-carboxy-heptadecanylamino) · butyryl mino] · Butyrylamino} -butyryl-, (S) -4-carboxy-4- «S} -4-carboxy-4- {2- [2- (2- {2 · [2- (2 - {(S) - 4-carboxy-4- [10- (4-carboxy-phenoxy) -decanoylamino] -butyrylamino).

5-ethoxy) -ethoxy] -acetylamino) -ethoxy) -ethoxy) -acetylamino} -butyryl-, (S) -4-carboxy-4 - {(S) -4-carboxy-4- [2- (2- { 2- [2- (2- {2 - [(S) -4carboxy-4- (7-carboxy-heptanoylamino) -butyrylamino] -ethoxy) -ethoxy) -acetylamino] -ethoxy} -ethoxy) -acetylamino] -butyrylamino } butyryl-, (S} -4-carboxy-4 - {(S} -4-ca rboxi-4- [2- (2- {2 - [2 - (2 - {2 - [(S} -4 -carboxy-4- (11-carboxy-undecanoylamino) -butyrylamino] ethoxy} -ethoxy) -acetylamino] -ethoxy} -ethoxy) -acetylamino] -butyrylamino} -butyryl-, (S) -4-carboxy-4 - {(S) -4-carboxy-4- [2- (2- {2- [2- (2 {2-¡(S) -4-carboxy-4- (13-carboxy-tridecanoylamino) -butyrylamino] -ethoxy} -ethoxy) -acetylamino] -ethoxy} -ethoxy) -acetylamino]

10-butyrylamino} -butyryl-, (S) -4-ca rboxi-4- {(S) -4-carboxy-4- [2 - (2 - {2 - [2 - (2 - {2 - [(8 ) -4-carboxy-4- (1 S-ca rboxypentadecanoylamino) -butyrylamino] -ethoxy) -ethoxy) -acetylamino] -ethoxy} -ethoxy) -acetylamino] -butyrylamino} -butyryl-y (S) -4carboxy-4 - {(S) -4-carboxy-4-y2- (2- {2-y2- (2- {2 - [(S} -4-carboxy-4- (19-carboxy-nonadecanoylamino) -butyrylamino ] -ethoxy} ethoxy) -acetylamino] -ethoxy) -ethoxy) -acetylamino] -butyrylamino) -butyryl-

More preferred are stereoisomers, particularly enantiomers of these groups, either S or R enantiomers.

15 The tennlno quot; Rquot; in Table 3 is meant to mean the binding site of -C (O) -R to the peptidyl skeleton, i.e., particularly the E-amino group of Lys.

Table 3

<dl><dt>structure </dt><dd>IUPAC first name </dd></dl>

<dl><dt>or,) l .-. / '-... / ~~,' ~ oe </dt><dd>(S) -4-Carboxy-4-hexadecanoylamino-buliril yE-x53 </dd></dl>

<dl><dt>or ~~ ~ '1ftI ~ C1 or </dt><dd>~ (S) 4-Carboxy-4-octadecanoylamino-butyryl yE-x70 </dd></dl>

<dl><dt>or ~: L ('/' - / ' </dt><dd>~ 4-Hexadecanoylamino-butyryl GABA-x53 </dd></dl>

<dl><dt>1. ~. </dt><dd>[ 1 I 11T ni i j J 4- (3- {(R) -2,5,7,8-Telramethyl-2 "4R, 8R) -4, 8,12 -trimethyl-tridecyl) -chroman-6-yloxycarbonyl) -propionylamino} -bubiryl GABA-x60 </dd></dl>

<dl><dt>OR </dt><dd /></dl>

<dl><dt>~~~ O </dt><dd>4-octadecanoylamino-butyryl GABA-x70 </dd></dl>

<dl><dt>OO Jlq-JL .............-........ / NI; gt; / .......... ~ / ~ </dt><dd /></dl>

<dl><dt>l ,, </dt><dd /></dl>

<dl><dt>quot; </dt><dd>4- «Z) -Octadec-9-enoylamino) -butyryl GABA-x74 </dd></dl>

<dl><dt>structure </dt><dd>IUPAC first name </dd></dl>

<dl><dt>/ 'gt; ! Ji Q c: r · 1 ;. ' l '/'- -ORO i, {/ -.... ~ /' '-' ~ o .. IL-lf ~ ./ Y 1: 'p ~~ O-lt;</dt><dd>6 - [(4,4-Diphenyl-cyclohexyloxy) -hydroxy-phosphoryloxy) -hexanoyl Fosfo1 </dd></dl>

<dl><dt>R y ~ ............... / O </dt><dd>Hexadecanoil 53 </dd></dl>

<dl><dt>oo • • • w ~, quot;, -, ~ .... R .... fquot; - quot; '/' · -'- quot; '/' · _ / '-..- quot; '' '' '' '/' ~ A ... O! 1 'IO' ~: J ~ l </dt><dd>(S) -4-Carboxy-4- (1 5-carboxypentadecanoylamine) -bubiril '52 </dd></dl>

<dl><dt>~~~ / y: p, quot; l ~ O ooo (] -l G!</dt><dd>(8) -4-Carboxy-4- {3- [3 - ((28.3R, 48.5R) -5-carboxy-2,3,4,5-tetrahydroxypentanoylamino) -propionylamino] -propionylamino} -butyryl yE-x59 </dd></dl>

<dl><dt>'~' quot; Cy ~ • o ~ v'- ~,. ~~~~~~~</dt><dd>(8) -4-Carboxy-4- {3- [(R) -2, 5, 7, 8-tetramethyl-2 - ({4 R, 8 R) -4, 8, 12 -trimethyl-tridecyl) chroman -6-ioxycarbonyl) -propionylamino} -bubiryl yE-x60 </dd></dl>

<dl><dt>~ "quot; OOI! ~ NH ~ quot; ~ O</dt><dd>(S) -4-Carboxy-4- ((92,122) -ocladeca-9, 12 -dienoylamino) -buli ril yE-x61 </dd></dl>

<dl><dt>structure </dt><dd>IUPAC first name </dd></dl>

<dl><dt>o Olt; Olt; or ~ .l ,, /, - ~~ Volt; • lt; lt; ~ o</dt><dd>(S} -4-Carboxy-4- [6 - ((2S, 3R, 4S, 5R) -5-carboxy-2,3,4,5-tetrahydroxypentanoylamino) -hexanoylamino) -butyryl yE-x64 </dd></dl>

<dl><dt>O CH CH O ~ y ~ ji R! 1, Y '; Y' ot ~ OO CH CH</dt><dd>(S} -4-Carboxy-4 - ((2S, 3RAS, 5R) -5-carboxy-2,3,4,5-tetrahydroxypentanoylamino) -butyryl yE-x65 </dd></dl>

<dl><dt>or ~ H, quot; ('- ~~.' ~ oo </dt><dd>(S} -4-Carboxy-4-tetradecanoylamino-butyryl yE-x69 </dd></dl>

<dl><dt>~~ '~, ~ o, .00 0 n </dt><dd>(S} -4- {11-Benzyloxycarbonyl-undecanoylamino) -4-carboxy-butyryl yE-x72 </dd></dl>

<dl><dt>~~~~ and V '~ ~ oo quot; . ! ~</dt><dd>(S} -4-Carboxy-4- [11 - ((2S, 3R, 4R, 5R) -2,3,4,5,6-pentahydroxyhexylcarbamoyl) undecanoylamino-butyryl yE-x73 </dd></dl>

<dl><dt>or ~ Nlt; ~. .TO. '\</dt><dd>(S} -4-Carboxy-4 - ((Z) -octadec-9-enoylamino} -butyryl yE-x74 </dd></dl>

<dl><dt>oo ~ A ---- yquot; ' ',' ~. / ',, Agt;</dt><dd>(S} -4-Carboxy-4- (4-dodecyloxy-benzoylamino) -butyryl yE-x75 </dd></dl>

<dl><dt>structure </dt><dd>IUPAC first name </dd></dl>

<dl><dt>~? '(' '-'''_ /, ~~ .., ..... quot; </dt><dd>(S) -4-Carboxy-4-henicosanoylamino-buliril VE-x76 </dd></dl>

<dl><dt>"Q Jl ~, quot; -</dt><dd>(8) -4-Carboxy-4-docosanoylamino-butyryl VE-x77 </dd></dl>

<dl><dt>(~ quot; ~ / 1,., Agt; u </dt><dd>(8) -4-Carboxy-4- «Z) -nonadec-1 O-enoylamino) -buti ril VE-x79 </dd></dl>

<dl><dt>r J- / Yquot;! vJ ..ru </dt><dd>(8) -4-Carboxy-4- (4-decyloxy-benzoylamino) -butyryl VE-x80 </dd></dl>

<dl><dt>Or Io ~~ quot; quot; p I, - 'R i I quot; quot; - ~ ./~o II d ~</dt><dd>(S) -4-Carboxy-4 - [(4'-octyloxy-biphenyl-4-carbonyl) -amino) -butyryl- VE-x81 </dd></dl>

<dl><dt>or Ú ~ 1: ~ ~ ~ ~ ~ .., Aa </dt><dd>(8) -4-Carboxy-4- (12-phenyl-dodecanoylamino) -butyryl VE-x82 </dd></dl>

<dl><dt>structure </dt><dd>IUPAC first name </dd></dl>

<dl><dt>~~ -quot; ~ 1 ~ :: 'I' </dt><dd>(S) -4-Carboxy-4-icosanoylamino-buliril yE-X95 </dd></dl>

<dl><dt>'and quot ;, ~, ~ O? 0lt; 0 o </dt><dd>(Sr4-Carboxy-4- «S) -4-carboxy-4-hexadecanoylamino-butyrylamino) -butyryl yE-yE-x53 </dd></dl>

<dl><dt>'' '' ('-1 _ ~~~~ (quot;) quot;' ~) -: gt; quot;</dt><dd>(S) -4-Carboxy-4- «S) -4-carboxy-4-octadecanoylamino-butyrylamino) -butyryl yE-yE-x70 </dd></dl>

<dl><dt>, '1 ~~~~~ quot; -',, or </dt><dd>3- (3-octadecanoylamino-propionylaminorpropionyl) B-Ala-B-Ala, 70 </dd></dl>

<dl><dt>'~ U ~. ~ Quot;, r ---./'-./'-./'-./'-./'- quot; {l ~) ~</dt><dd>3- (3-Hexadecanoylamino-propionylamino) -propionyl p-Ala-p-Ala, 53 </dd></dl>

<dl><dt>H ~ / 'O o </dt><dd>3-Hexadecanoylamino-propionyl p-Ala-x53 </dd></dl>

<dl><dt>~ l-0- ~, ~ O ~ 00 'H' OH </dt><dd>(Sr4-Carboxy-4 - [(R) -4- "3R, 5S, 7R, 8R, 9R, 1 OS, 12S, 13R, 14R, 17R) -3,7,12-Hydroxy-B, 1 O, 13- trimethyl-hexadecahydro-cyclopenta [ajfenantren-17-yl pentanoylaminoj-butyryl- yE-x16 </dd></dl>

<dl><dt>or 0quot; /,-1. U, ¡'' quot ;. '~ NH: ~ O ~ I CH</dt><dd>(S) -4-Carboxy-4 - [(R) -4- "3R, 5R, 8R, 9S, 1 OS, 13R, 14S, 17R) -3-hydroxy-1 O, 13-dimethyl-hexadecahydrocyclopenta [ajfenantren-17 -yl rpentanoylamino j-butyryl yE-x19 </dd></dl>

<dl><dt>structure </dt><dd>IUPAC first name </dd></dl>

<dl><dt>~ 'quot; .c ••: Ji ¡. , llt; Tquot; '~ o</dt><dd>(8) -4-Carboxy-4- "98, 1 OR) -9, 1O, 16-trihydroxy-hexadecanoylamino) -butyry 1- yE-x25 </dd></dl>

<dl><dt>and ~~ O </dt><dd>Tetradecanoil '69 </dd></dl>

<dl><dt>OY Olt; OR</dt><dd>11-Carboxi-undecanoil 71 </dd></dl>

<dl><dt>/ '~~~~~~~~ Jo [~ O, 1) </dt><dd>11-Benzyloxycarbonyl undecanoyl , 72 </dd></dl>

<dl><dt>yJ; 0 '~ quot; , UT ¡or Il cfquot; O o</dt><dd>(S) -4-Carboxy-4-S) -4-carboxy-4-tetradecanoylamino-butyrylamino) -butyryl yE-yE-x69 </dd></dl>

<dl><dt>ffd 'OOI ¿~~ quot; ¡YI Olt; </dt><dd>6- [Hydroxy- (naphthalen-2-yloxy) -phosphoryloxy] -hexanoyl Fosfo2 </dd></dl>

<dl><dt>Or ~ a + ~ I quot; CH</dt><dd>6- [Hydroxy- (5-phenyl-pentyloxy) -phosphoryloxy] -hexanoyl Fosfo3 </dd></dl>

structure

~ o

I

OR

cxjO ~

IHO

"-quot;

0: 5

~; ro

quot; quot; 'quot;

! I

TO

. ~~~~ Xy

'~~ 0 (' - '~~

· V · ~

! 1, '' quot; '' '......., J .. ~ ........- Y-, J ... ..

. ! . ') ..:.

lY quot;

quot; . '~~

I! . lt;.,

, ......

1quot; yquot; . {quot; yquot;

. ~

..

1 1 ~

IUPAC

4- (Naphthalene-2-sulfonylamino) -4-oxo-butyryl

4- (Biphenyl-4-sulfonylamino) -4-oxo-butyryl

(8) -4-Carboxy-4- {(8) -4-carboxy-4- [2 - (2- {2- [2- (2- {2 - [{8) -4-carboxy-4-} {17 carboxy-heptadecanoylamino) -butyrylamino) -ethoxy} -ethoxy) -acetylamino] -ethoxy} ethoxy) -acetylamino] -butyrylamino} -butyryl

(8) -4-Carboxy-4- [2 - (2 - {2- [2 - (2 - {2 - [(8) -4-carboxy-4- (17 -ca-rboxiheptadecanoylamino) -butyrylamino] -eloxi } -eloxi) -acetylamino] -ethoxy} ethoxy) acetylamino] -butyryl

(8) -4-Carboxy-2 - {(8) -4-carboxy-2- [2- (2- (2- [2 - {2 - (2 - [(8) -4-carboxy-4-) (1 7carboxy-heptadecanoylamino) -butyrylamino) -ethoxy} -ethoxy) -acetylamino] -ethoxy} ethoxy) -acetylamino] -butyrylamino} -butyryl

(S) -4-Carboxy-2 - [2- (2 - {2 - [2- (2- {2 - [(S) -4-carboxy-4- (1 7 -ca rboxiheptadecanoylamino) -butyrylamino] - ethoxy} -ethoxy) -acetylamino] -ethoxy} -ethoxy) acetylamino) -butyryl

(8) -4-Carboxy-4 {(8) -4-carboxy-4- [2 - (2 - {2 - [(8) -4-carboxy-4- (1 7 -carboxyhepdecanelamino) -bubyrylamino] - ethoxy} -ethoxy) -acetylamino-bulyrylamino} bulyryl

first name

Sulfonamide

Sulfonamide 2

x100

x101

x102

x103

04

structure

, gt; --lquot; '-' '' »~

0¡ ~. ~~

and '' '' quot; gt; quot; 1.¡, ~~ il-l_.

TZ '

1 quot;

quot; vquot;

J 'YOgt; ~ '

'~ ~

1, quot; ~ quot; ''; l

, quot;

"Quot;" quot; quot;

~

. ~~~ '~

I

,, - 1y quot;

~~;) ~ quot;

! 'fr

JY

') ~ quot; ~, ~~~

,, ---. or "P and O")

': c ~~ /' _-. Jquot; /, / y ,,,, ~~. ~ and

1 "Yquot; ,

¡•! - ~ '- /' '- and-,' '

~

0 ~~

J "

0 '

IUPAC

(8) -4-Carboxy-4- [2 - (2- {2 - [(8) -4-carboxy-4- {17 -carboxy-heptadecanoylamino) butyrylamino] -ethoxy} -ethoxy) -acetylamino] -butyryl

(8) -4-Carboxy-2 [(8) -4-carboxy-2 - [2- (2- {2 - [(8) -4-carboxy-4- (17-carboxyheptadecanoylamino) -butyrylamino) -ethoxy } -ethoxy) -acetylamino) -butyrylamino} butyryl

(8) -4-Carboxy-2 - [2 - (2 - {2 - [(8) -4-carboxy-4- (17 -carboxy-heptadecanoylamino) butyrylamino] -ethoxy} -ethoxy) -acetylamino) -butyryl

2- (2 - {2 - [2 - {2- {2 - [(8) -4-Carboxy-4- (1 7 -carboxy-heptadecanoylamino) butyrylamino] -ethoxy} -ethoxy) -acetylamino] -ethoxy } -ethoxy) -acetyl

2- (2 - {2 - [(8) -4-Carboxy-4- (1 7 -carboxy-heptadecanoylamino) -butyrylamino-ethoxy} -ethoxy) -acetyl

(8) -4-Carboxy-4- ((8) -4-carboxy-4 {(8) -4-carboxy-4- [(8) -4-carboxy-4- (19-carboxy-nonadecanoylamino) -butyrylamino] -butyrylamino} -butyrylamino) -butyryl

2- (2- {2 - [2 - (2 - {2 - {(8) -4-Carboxi-4- (16-1 H-tetrazol-5-yl-hexadecanoylamino) butyrylamino] -ethoxy} -ethoxy) -acetylamino) -ethoxy} -ethoxy) -acetyl

2- (2- {2 - [2 - (2 - {2 - [(8) -4-Carboxy-4- (16-carboxy-hexadecanoylamino) butyrylamino] -ethoxy} ethoxy) -acetylamino) -ethoxy} -ethoxy )-acetyl

(8) -4-Carboxy-4- {(8) -4-carboxy-4 - [(8) -4-carboxy-4- (1 7 -carboxyheptadecanoylamino) -butyrylamino) -butyrylamino} -butyrylO

first name

x105

x106

x107

x10B

x109

x110

x111

x112

x113

structure

. ' ~ ,. 'v'O

_~~~ , -quot; J ' ~, ••

p ',. .

'quot; lt;: Ivlt; quot; ,,;, ..... -.; ..

'Y' '' '-_' '' A / '' '' Y ~~~

-quot;

quot; and ...

1.Y '. j

quot; .

,

. ' ~ quot; ~ -'I '

'and' 'and quot;

~~~~ ,,,, - ,, ~ ,, - ,,, ~, ~.

., or,

w. ~.

4y ~ ~ V '

~~~~~

quot; .0,. ,.,

'~~~~~ I:' ~

IUPAC

(5) -4-Carboxy-4 - ((5) -4-carboxy-4- {2- [2- (2- {2- [2- (2 - {(5) -4-carboxy-4-) [10- (4-carboxy-phenoxy) -decanoylamino] -butyrylamino} -α-toxy) -ethoxy] -acetylamino} -ethoxy) ethoxy) -acetylamino} -butyryl

(S) -4-Carboxy-4 - {(S) -4-carboxy-4- [2- (2- {2- [2- (2- {2 - [(S) -4-carboxy-4-) (7-carboxyheptanoylamino) -butyrylamino] -ethoxy) ethoxy) -acetylamino) -ethoxy} -ethoxy) acetylamino) -bubyrylamino} -butyryl

(S) -4-Carboxy-4 - {(S) -4-carboxy-4- [2- (2- {2- [2- (2- {2 - [(S) -4-carboxy-4-) (11-carboxy-undecanoylamino) -butyrylamino] -ethoxy} -ethoxy) -acetylamino] -ethoxy) ethoxy) -acetylamino] -butyrylamino} -butyryl

(S) -4-Carboxy-4 - {(S) -4-carboxy-4- [2- (2- {2- [2- (2- (2 - [(S) -4-carboxy-4-) (13carboxytridecanoylamino) -butyrylamino] -ethoxy} -ethoxy) -acetylamino] -ethoxy} -ethoxy) acetylamino) -butyrylamino} -butyryl

(S) -4-Carboxy-4 - {(S) -4-carboxy-4- [2- (2- {2- [2- (2- {2 - [(S) -4-carboxy-4-) (15carboxy-pentadecanoylamino) -bubyrylamino] -eloxy} -ethoxy) -acetylamino] -ethoxy) ethoxy) -acetylamino] -butyrylamino} -butyryl

(S) -4-Carboxy-4 - {(S) -4-carboxy-4- [2- (2- {2- [2- (2- {2 - [(S) -4-carboxy-4-) (1-carboxy-nonadecanoylamino) -butyrylamino] -ethoxy} -ethoxy) -acetylamino] -ethoxy) ethoxy) -acetylamino) -butyrylamino} -butyryl

first name

x11 4

x11 5

x116

x117

x11 8

x119 5

In some embodiments, the invention relates to peplide compounds of formula (1) as defined above, wherein X 14 represents an amino acid residue selected from Lys, Qrn, Oab and Dap, wherein the side chain group - NH2 is functionalized by -C (Q} -R5, X40 represents an amino acid residue selected from Lys, Qrn, Dab and Dap, in which the side chain group -NH2 can be functionalized by _C (Q) -R5, and R5 is a lipophilic moiety selected from a straight or branched acyclic saturated or unsaturated hydrocarbon (C4-C30) group, and / or a saturated, unsaturated or cyclic aromatic group, in which the lipophilic moiety can be attached to the NH2 side chain group by a connector selected from (~ -Ala) 1-4, (v-Gluh4, (E-Ahx) 1-4 or (GASA) l -4 in all stereoisomeric forms.

In certain embodiments, X14 represents an amino acid residue with a side chain group -NH2 functionalized, such as Lys, Qrn, Oab or functionalized Dap, wherein at least one H atom of the side chain group -NH2 is substituted by _C (Q) -R5, which is selected from the group consisting of the substituents according to Table 3 above

In some embodiments, X14 represents an amino acid residue selected from Lys, Qrn, Dab and Dap, wherein the side chain group -NH2 is functionalized by _C (Q) _R5, and _C (Q} -R5 is selected from the group which consists of the substituents according to Table 3 above.

In some embodiments of the invention, the X14 and / or X40 position in formula (11) represents Lysine (Lys). Lys at position 14 is functionalized with a group _C (Q) R5 as described above. In other embodiments, X40 is absent and X14 is Lys functionalized with _C (Q) -R5, wherein R5 is as defined above. In particular, X14 is Lys functionalized with C (Q) _R5, which is selected from the group consisting of (S) -4-carboxy-4

hexadecanoylamino-butyryl (VE-xS3), (S) -4-carboxy-4-octadecanoylamino-butyryl (VE-x70), 4-hexadecanoylaminobutyryl (GASA-xS3), 4- {3 - [(R) -2, 5 , 7, 8-tetramethyl-2- "4R, 8R) -4, 8,12 -trimethyl-tridecyl) -chroman-6-yloxycarbonyl] propionylamino} -butyryl- (GABA-x60), 4-ocladecanoylamino-butyryl ( GABA-x70), 4- "Z} -octadec-9-enoylamino) -butyryl (GABA-x 7 4), 6 - [(4,4-diphenyl-cyclohexyloxy) -h-idroxy-phosphoryloxy] -hexa-noyl (Phospho1 ), hexadecanoyl (xS3), (S) -4-ca rboxi-4 (1S-carboxy-pentadecan oylamino) -butyryl (xS2), (S) -4-ca rboxi-4- {3- [3- (( 2S, 3R, 4S, SR) -S-carboxy-2, 3,4, S-tetrahydroxypentanoylamino) -propionylamino] -propionylamino} -butyryl (VE-x59), (5) -4-carboxy-4- { 3-J (R) -2,5,7, 8-te tramethyl-2 "4R, 8R) -4, 8, 12 -trimethyl-tridecyl) -chroman-6-yloxycarbonyl] -propionylamino} -buyl (VE-x60), (5) -4-ca rboxi-4- ((9Z, 12Z) octadeca-9,12-d ienoilamno) -butyryl (VE-x61), (S) -4-ca rboxi-4- [6- «2S, 3R, 4S, SR) -S-carboxy-2, 3,4, S- tetrahydroxypentanoylamino) -hexanoylamino] -butyryl (yE-x64), (8) -4-ca-rboxi-4- ((28.3R, 48, SR) -S-ca rboxi-2,3, 4, S- tetrahydroxypentanoylamino) -butyryl (VE-x6S), (8) -4.-carboxy-4-tetrad ecanoylamino-butyryl (VE-x69), (8) -4- (11-benzyloxycarbonylundecanoylamino) -4-ca rboxi-butyryl (VE-x 72), (5) -4-ca rboxi-4- [11- ((25, 3R, 4R, 5R) -2, 3,4,5, 6-penta hydroxyhexylcarbamoyl) -undecanoylamino ) -butyryl (VE-x 73), (S) -4-carboxy-4- 'Z) -octadec-9-enoylamino) -butyryl (VE-x 7 4), (S) 4-carboxy-4- (4-dodecyloxy-benzoylamino) -butyryl (VE-x 7 5), (S} -4-carboxy-4-henicosanoylamino-butyryl (VE-x76), (S} -4carboxy-4-docosa noilam-n-butyryl (VE-x 77), (S} -4-carboxy-4- "Z) -nonadec-1 O-enoylamino) -butyryl (VE-x79),(S} -4-carboxy-4- (4-decyloxy-benzoyl-mino) -butyryl (VE-x80), (S) -4-carboxy-4 - [(4'-cyloxybiphenyl-4-carbonyl) -amino) - butyryl (VEx81), (S) -4-carboxy-4- (12-phenyl-dodecanoylamino) -butyryl (VE-x82), (S) -4-carboxy-4-ycosanoylamino-butyryl (VE-x9S), ( S) -4-carboxy-4- «S) -4-carboxy-4-hexadecanoylamino-butyrylamino) -butyryl (VE-VE-xS3), (S) -4-carboxy-4-« S) -4-carboxy4 -octadecanoylamino-butyrylamino) -butyryl (VE-VE-x70) and 3- (3-ocladecanoylamino-propionylamino) -propionyl (p-Ala-pAla-x70)(S) -4-carboxy-4- «S) -4-carboxy-4-octadecanoylamino-butyrylamino) -butyryl (VE-VE-x70) and 3- (3-ocladecanoylamino-propionylamino) -propionyl (p-Ala-pAla- x70)(S) -4-carboxy-4- «S) -4-carboxy-4-octadecanoylamino-butyrylamino) -butyryl (VE-VE-x70) and 3- (3-ocladecanoylamino-propionylamino) -propionyl (p-Ala-pAla- x70)

In some embodiments, X14 is Lys functionalized with C {Q) _R5, which is selected from the group consisting of (S) -4-carboxy-4-hexadecanoylamino-butyryl (VE-xS3) and (S) -4-carboxy- 4-octadecanoylamino-butyryl (VE-x70).

A further embodiment refers to a group of compounds, wherein

R1

It's NH2,

R2 is NH2o

R1

and R2 are NH2.

It also reveals a group of compounds, in which

X3 represents an amino acid residue selected from Gln, Glu and His,

X12 represents an amino acid residue selected from lIe and Lys,

X14 represents an amino acid residue that fills a side chain with a group -NH2, wherein the side chain group -NH2 is driven by _C (Q) _R5, wherein R5 is as described above,

X 15 represents an amino acid residue selected from Asp and Glu,

X16 represents an amino acid residue selected from Ser, Lys, Glu and Gln,

X17 represents an amino acid residue selected from Arg, Lys, Glu, Ie, Gln, Leu, Aib, Tyr and Ala,

X18 represents an amino acid residue selected from Ala, Arg, Aib, Leu, Lys and Tyr, X19 represents an amino acid residue selected from Ala, Gln, Val and Aib,

X20 represents an amino acid residue selected from Gln, Aib, Phe, Arg, Leu, Lys and His,

X21 represents an amino acid residue selected from Asp, Glu, Tyr and Leu,

X28 represents an amino acid residue selected from Asn, Ala, Aib, Arg and Lys,

X29 represents an amino acid residue selected from Gly, Thr, Aib, D-Ala and Ala,

X40 is either absent or represents Lys. A further embodiment refers to a group of compounds, wherein

X3 represents an amino acid residue selected from Gln, Glu and His,

X12 represents an amino acid residue selected from lIe and Lys,

X14 represents an amino acid residue having a side chain with a -NH2 group, in which the group of

side chain -NH2 is functionalized by -C (O) -R5, wherein R5 is as described above,

X15 represents an amino acid residue selected from Asp and Glu,

X16 represents an amino acid residue selected from Ser, Lys, Glu and Gln,

X17 represents an amino acid residue selected from Arg, Lys, Glu, Gln, Leu, Aib, Tyr and Ala,

X18 represents an amino acid residue selected from Ala, Arg, Aib, Leu and Tyr,

X19 represents an amino acid residue selected from Ala, Val and Aib,

X20 represents an amino acid residue selected from Pip, (S) MeLys, (R) MeLys and (S) MeOrn,

X21 represents an amino acid residue selected from Asp, Glu and Leu,

X28 represents an amino acid residue selected from Asn, Ala, Aib and Ser,

X29 represents an amino acid residue selected from Gly, Thr, Aib, D-Ala and Ala,

X40 is either absent or represents Lys A further embodiment refers to a group of compounds, wherein

X3 represents an amino acid residue selected from Gln, Glu and His,

X12 represents ne,

X14 represents an amino acid residue having a side chain with a -NH2 group, in which the group of

side chain -NH2 is functionalized by _C (O) _R5, wherein R5 is as described above,

X15 represents an amino acid residue selected from Asp and Glu,

X16 represents an amino acid residue selected from Ser, Lys, Glu and Gln,

X17 represents an amino acid residue selected from Arg, Lys, Glu, Gln, Leu, Aib, Tyr and Ala,

X18 represents an amino acid residue selected from Ala and Arg,

X19 represents an amino acid residue selected from Ala and Val,

X20 represents an amino acid residue selected from Pip, (S) MeLys, (R) MeLys and (S) MeOrn,

X21 represents an amino acid residue selected from Asp, Glu and Leu,

X28 represents an amino acid residue selected from Asn and Ala,

X29 represents an amino acid residue selected from Gly, Thr and O-Ala,

X40 is either absent or represents Lys It also reveals a group of compounds, in which

X3 represents an amino acid residue selected from Gln, Glu and His, X12 represents an amino acid residue selected from Ile and Lys, X14 represents an amino acid residue having a side chain with a -NH2 group, in which the group of

side chain -NH2 is functionalized by _C (Q) _Rs, wherein RS is as described above, X15 represents an amino acid residue selected from Asp and Glu, X16 represents an amino acid residue selected from Ser, Lys, Glu and Gln, X17 represents an amino acid residue selected from Arg, Lys, Glu, Gln, Leu, Aib, Tyr and Ala, X18 represents an amino acid residue selected from Ala and Arg, X19 represents an amino acid residue selected from Ala and Val, X20 represents an amino acid residue selected from Gln, Aib, Lys and His, X21 represents an amino acid residue selected from Asp, Glu and Leu, X28 represents an amino acid residue selected from Asn and Ala, X29 represents an amino acid residue selected from Gly, Thr and O-Ala, X40 is either absent or represents Lys

A group of compounds is also disclosed, in which X3 represents an amino acid residue selected from G! N and Glu, X 12 represents Ile, X14 represents Lys, in which the side chain group -NH2 is functionalized by one of the groups

selected from (S) -4-carboxy-4-hexadecanoylamino-butyryl-, (S) -4-carboxy-4-octadecanoylamino-butyryl-, (S} -4-carboxy-4-S) -4-carboxy-4-octadecanoylamino-butyrylamino) -butyryl-, 3- (3-octadecanoylamino-propionylamino} propionyl- and 4-octadecanoylamino-butyryl-, (S) -4-ca rboxi-4-henicosanoylamino-butyryl-,

X15 represents an amino acid residue selected from Glu and Asp, X16 represents an amino acid residue selected from Ser and Lys, X17 represents Arg, X18 represents Ala, X19 represents Ala, X20 represents an amino acid residue selected from Gln and Aib, X21 represents an amino acid residue selected from Asp and Glu, X28 represents an amino acid residue selected from Asn and Ala, X29 represents an amino acid residue selected from Gly and Thr, X40 is absent

A group of compounds is also disclosed, in which X3 represents Glu, X 12 represents lIe, X14 represents Lys, in which the side chain group -NH2 is functionalized by one of the groups

selected from (S) -4-carboxy-4-hexadecanoylamino-butyryl-, (S) -4-carboxy-4-octadecanoylamino-butyryl-, (S} -4-carboxy-4-S) -4-carboxy-4-octadecanoylamino-butyrylamino) -butyryl-, 3- (3-octadecanoylamino-propionylamino} propionyl- and 4-octadecanoylamino-butyryl-, (8) -4-caboxy-4-henicosanoylamino-butyryl-,

X15 represents an amino acid residue selected from Glu and Asp, X16 represents a remainder of the ammount selected from Ser and Lys,

X17 represents Arg, X18 represents Ala, X19 represents Ala, X20 represents an amino acid residue selected from Gln and Aib, X21 represents an amino acid residue selected from Asp and Glu, X28 represents an amino acid residue selected from Asn and Ala, X29 represents an amino acid residue selected from Gly and Thr, X40 is absent.

A group of compounds is also disclosed, in which X3 represents Gln, X12 represents lIe, X14 represents Lys, in which the side chain group -NH2 is functionalized by one of the groups

selected from (5) -4-ca rboxi-4-hexadecanoilami non-butyryl-, (5) -4-carboxy-4-octadecanoila m ino-buliryl-, (5) -4carboxi-4- «5) -4- carboxy-4-octadecanoylamino-butyrylamino) -butyryl-, 3- (3-octadecanoylamino-propionylamino) propionyl- and 4-ocladecanoylamino-butyryl-, (5) -4-carboxy-4-henicosanoylamino-butyryl-,

X15 represents an amino acid residue selected from Glu and Asp, X16 represents an amino acid residue selected from Ser and Lys, X17 represents Arg,

X18 represents Ala, X19 represents Ala, X20 represents an amino acid residue selected from Gln and Aib, X21 represents an amino acid residue selected from Asp and Glu, X28 represents an amino acid residue selected from Asn and Ala, X29 represents an amino acid residue selected from Gly and Thr, X40 is absent

A further embodiment refers to a group of compounds, wherein X14 represents Lys, wherein the side chain group -NH2 is functionalized by one of the selected groups of (5) -4-carboxy-4-hexadecanoylamino-butyryl -, (5) -4-carboxy-4-ocladecanoylamino-butyryl-, 4

octadecanoylamino-butyryl-, hexadecanoyl-, (5) -4-carboxy-4-henicosa noilam-n-butyryl-, (5) -4-ca-rboxi-4- ((5) -4-ca-rboxi-4-octadecanoylamino-butyrylamino) -butyryl-, 3- (3-octadecanoylamino-propionylamino) -propionyl-. A further embodiment refers to a group of compounds, in which X14 represents Lys, in which the side chain group -NH2 is functionalized by one of the groups

selected from (5) -4-carboxy-4-octadecanoylamino-butyryl-, 4-octadecanoylamino-butyryl-, (5) -4-carboxy-4-hydrogenicosaminyl-butyryl-, (5) -4-carboxy-4- «5) 4-carboxy-4-octadecanoylamino-butyrylamino) -butyryl-, 3- (3-octadecanoylamino-propionylamino) -propionyl-

A further embodiment refers to a group of compounds, wherein

X14 represents Lys, in which the side chain group -NH2 is functionalized by one of the selected groups of (S) -4-carboxy-4-hexadecanoylamino-butyryl-, (S) -4-carboxy-4-octadecanoylamino- butyryl-. It also reveals a group of compounds, in which

X3 represents an amino acid residue selected from Gln and Glu, X12 represents lIe,

X14 represents Lys, in which the side chain group -NH2 is functionalized by one of the groups

selected from (S) -4-carboxy-4-hexadecanoylamino-butyryl-y (S) -4-carboxy-4-octadecanoylamino-butyryl-,

X15 represents an amino acid residue selected from Glu and Asp,

X16 represents an amino acid residue selected from Ser and Lys,

X17 represents Arg,

X18 represents Ala,

X19 represents Ala,

X20 represents an amino acid residue selected from Gln and Aib,

X21 represents an amino acid residue selected from Asp and Glu,

X28 represents a remainder of the ammount selected from Asn and Ala,

X29 represents an amino acid residue selected from Gly and Thr,

X40 is absent A group of compounds is also revealed, in which

X3 represents an amino acid residue selected from Gln, His and Glu,

X 12 represents ne, X14 represents Lys, in which the side chain group -NH, is functionalized by one of the groups selected from (S) -4-carboxy-4-hexadecanoylamino-butyryl-y (S) -4-carboxy-4-octadecanoylamino-butyryl-,

X15 represents Glu,

X16 represents an amino acid residue selected from Glu and Lys,

X17 represents Glu,

X18 represents Ala,

X19 represents Va l,

X20 represents Arg,

X21 represents Leu,

X28 represents an amino acid residue selected from Asn, Aib and Ala,

X29 represents an amino acid residue selected from Gly and Thr,

X40 is absent. It also reveals a group of compounds, in which

X3 represents Glu,

X 12 represents lIe,

X14 represents Lys, in which the side chain group -NH2 is functionalized by one of the groups

selected from (S) -4-carboxy-4-hexadecanoylamino-butyryl-y (S) -4-carboxy-4-octadecanoylamino-butyryl-, X15 represents Glu, X16 represents an amino acid residue selected from Glu and Lys, X17 represents Glu, X18 represents Ala, X19 represents Val,

X20 represents Arg, X21 represents Leu,

X28 represents an amino acid residue selected from Asn, Aib and Ata,

X29 represents Gly,

X40 is absent. A further embodiment refers to a group of compounds, wherein

X3 represents an amino acid residue selected from Gln, His and Gtu,

X12 represents an amino acid residue selected from lIe and Lys,

X14 represents Lys, in which the side chain group -NH2 is functionalized by one of the groups

selected from (S) -4-carboxy-4-hexadecanoylamino-butyryl-y (S) -4-carboxy-4-octadecanoylamino-butyryl-,

X15 represents an amino acid residue selected from Glu and Asp,

X16 represents Glu,

X17 represents an amino acid residue selected from Arg and Gln,

X18 represents an amino acid residue selected from Ala and Arg,

X19 represents Ala,

X20 represents an amino acid residue selected from Pip, (S) MeLys, {R) MeLys and (S) MeOm,

X21 represents Glu,

X28 represents an amino acid residue selected from Asn, Ser and Ala,

X29 represents an amino acid residue selected from Gly and Thr,

X40 is absent A group of compounds is also revealed, in which

X3 represents an amino acid residue selected from Gln, His and Glu,

X12 represents a remainder of the ammount selected from lIe and Lys,

X14 represents Lys, in which the side chain group -NH2 is functionalized by one of the groups

selected from (S) -4-carboxy-4-hexadecanoylamino-butyryl-, hexadecanoyl- and (S) -4-carboxy-4

octadecanoylamino-butyryl-,

X15 represents an amino acid residue selected from Glu and Asp,

X16 represents an amino acid residue selected from Ser, Lys, Glu and Gln,

X17 represents an amino acid residue selected from Arg, Leu, Aib, Tyr, Glu, Ala and Lys,

X18 represents an amino acid residue selected from Ala, Aib, Leu and Tyr,

X19 represents an amino acid residue selected from Ala, Val and Aib,

X20 represents Aib,

X21 represents an amino acid residue selected from Glu, Leu and Tyr,

X28 represents an amino acid residue selected from Asn, Arg and Ala,

X29 represents an amino acid residue selected from Gly, Ala, D-Ala and Thr,

X40 is either absent or represents Lys. It also reveals a group of compounds, in which

X3 represents an amino acid residue selected from Gln, His and Glu,

X12 represents an amino acid residue selected from lIe and Lys,

X14 represents Lys, in which the side chain group -NH2 is functionalized by one of the groups

selected from (S) -4-carboxy-4-hexadecanoylamino-butyryl-y (S) -4-carboxy-4-octadecanoylamino-butyryl-,

X15 represents an amino acid residue selected from Glu and Asp,

X16 represents an amino acid residue selected from Ser, Lys and Glu,

X17 represents an amino acid residue selected from Arg, Lys, lIe, Glu and Gln,

X18 represents an amino acid residue selected from Ala, Arg and Lys,

X19 represents an amino acid residue selected from Ala, Val and Gln,

X20 represents an amino acid residue selected from Gln, Phe, Leu, Lys, His and Arg,

X21 represents an amino acid residue selected from Glu, Asp and Leu,

X28 represents a remainder of aminoacid selected from Asn, Arg, Lys and Ala,

X29 represents an amino acid residue selected from Gly, Aib and Thr,

X40 is either absent or represents Lys A further embodiment refers to a group of compounds, wherein

X 12 represents Ie A further embodiment refers to a group of compounds, wherein

X19 represents Ala. A further embodiment refers to a group of compounds, wherein

X16 represents Glu,

X20 represents an amino acid residue selected from Pip, (S) MeLys, (R) MeLys and (S) MeOrn. A further embodiment refers to a group of compounds, wherein

X28 represents Ala,

X29 represents Gly. A further embodiment refers to a group of compounds, wherein

X28 represents Asn,

X29 represents Thr. A further embodiment refers to a group of compounds, wherein

X3 represents an amino acid residue selected from Gln and Glu,

X12 represents an amino acid residue selected from lIe and Lys, X14 represents Lys, in which the side chain group -NH2 is functionally raised by _C (O) -R5, which is selected from (S) -4-carboxy-4-hexadecanoylamino-butyryl- (yE-x53) and (S) -4-carboxy-4-octadecanoylamino butyryl- (yE-x70),

X15 represents an amino acid residue selected from Asp and Glu,

X16 represents Glu,

X17 represents an amino acid residue selected from Arg and Gln,

X18 represents an amino acid residue selected from Ala and Arg,

X19 represents Ala,

X20 represents an amino acid residue selected from Pip, (S) -MeLys, (R) -MeLys, and (S) -MeOrn,

X21 represents Glu, X28 represents an amino acid residue selected from Asn, Ala and Ser,

X29 represents an amino acid residue selected from Gly and Thr,

X40 is absent.

Specific examples of peplid compounds of formula (1) are the compounds of SEO ID NO '8-16, in addition to salts and solvates thereof.

Specific examples of peptide compounds of formula (1) are the compounds of SEQ ID NO: 8-13 and 15, in addition to salts and solvates thereof.

In certain embodiments, that is, when the compound of formula (1) comprises genetically encoded amino acid residues, the invention further provides a nucleic acid (which may be DNA or RNA) encoding said compound, an expression vector comprising an acid nucleic such and a host cell containing a nucleic acid or expression vector such

In another aspect, the present invention provides a composition comprising a compound of the invention in admixture with a carrier. In preferred embodiments, the composition is a pharmaceutically acceptable composition and the carrier is a pharmaceutically acceptable carrier. The compound of the invention may be in the form of a salt, for example, a pharmaceutically acceptable salt or a solvate, for example, a hydrate. In yet another aspect, the present invention provides a composition for use in a method of medical treatment, particularly in human medicine.

In certain embodiments, the nucleic acid or expression vector can be used as therapeutic agents, for example, in gene therapy

The compounds of formula (1) are suitable for therapeutic application without an additional therapeutically effective agent. In other embodiments, however, the compounds are used in conjunction with at least one additional therapeutically active agent, as described in "Combination Therapy".

The compounds of formula (1) are particularly suitable for the treatment or prevention of diseases

or disorders caused by, associated with and / or accompanied by alterations in the metabolism of carbohydrates and / or lipids, for example, for the treatment or prevention of hyperglycemia, type 2 diabetes, glucose intolerance, type diabetes 1, obesity and metabolic syndrome. In addition, the compounds of the invention are particularly suitable for the treatment or prevention of degenerative diseases, particularly neurodegenerative diseases.

The disclosed compounds are used, among others, in preventing weight gain or promoting weight loss. Why "prevent"; it is indicated to inhibit or reduce when compared to the absence of treatment, and does not necessarily mean that it implies the complete cessation of a disorder

The compounds of the invention can produce a decrease in food intake and / or increase energy expenditure, producing the effect observed on body weight.

Regardless of their effect on body weight, the compounds of the invention can have a beneficial effect on circulating cholesterol levels, being able to improve lipid levels, particularly LDL, in addition to HOL levels (for example, increasing the HDlILOL relation)

Thus, the compounds of the invention can be used for the direct or indirect therapy of any condition caused

or characterized by excess body weight, such as the treatment and / or prevention of obesity, morbid obesity, inflammation linked to obesity, gallbladder disease linked to obesity, sleep apnea induced by obesity. They can also be used for the treatment and prevention of metabolic syndrome, diabetes, hypertension, atherogenic dyslipidemia, atherosisrosis, arteriosclerosis, coronary heart disease or cerebrovascular accident. Its effects on these conditions may be the result of or associated with its effect on body weight, or it may be independent of the same

Preferred medical uses include delaying or preventing the progression of the disease in type 2 diabetes, treating metabolic syndrome, treating obesity or preventing overweight, to decrease food intake, increase energy expenditure, reduce body weight, delay the progression of glucose intolerance (IGT) to type 2 diabetes; delay the progression from type 2 diabetes to diabetes requiring insulin; regulate the appetite; induce satiety; prevent re-fattening after a satisfactory weight loss; treat a disease or condition related to overweight or obesity; treat bulimia; treat binge eating; treat atherosclerosis, hypertension, type 2 diabetes, IGT, dyslipidemia, coronary heart disease, hepatic steatosis, treatment of beta-blocker poisoning,

Other preferred medical uses include the treatment or prevention of degenerative disorders, particularly neurodegenerative disorders such as Alzheimer's disease, Parkinson's disease,

Huntington, ataxia, for example, spinocerebellar ataxia, Kennedy's disease, myotonic dystrophy, dementia due to Lewy bodies, multi-systemic atrophy, amyotrophic lateral sclerosis, primary lateral sclerosis, spinal muscular atrophy, prion-associated diseases, for example, Creutzfeldt-Jacob, multiple sclerosis, telangiectasia, Batlen's disease, corticobasal degeneration, subacute combined degeneration of the spinal cord, tabes dorsalis, Tay-Sachs disease, toxic encephalopathy, infantile Refsum's disease, Refsum's disease, neuroacanthocytosis, Niemann-Pick, Lyme disease, MachadoJoseph's disease, Sandhoff's disease, Shy-Drager syndrome, wobbly hedgehog syndrome, proteopathy, cerebral-amyloid-angiopathy,degeneration of retinal ganglionic cells in glaucoma, synucleinopathies, tauopathies, frontotemporal lobar degeneration (FTLD), dementia, cadasil syndrome, hereditary cerebral hemorrhage with 10 amyloidosis, Alexander's disease, sepianopathies, familial amyloidic neuropathy, senile systemic amyloidosis, serpinopathies, AL amyloidosis (light chain) (primary systemic amyloidosis), amyloidosis AH (heavy chain), AA amyloidosis (secondary), medial aortic amyloidosis, ApoAI amyloidosis, ApoAII amyloidosis, ApoAIV amyloidosis, familial Finnish amyloidosis (FAF), lysozyme amyloidosis , amyloidosis by fibrinogen, amyloidosis by dialysis, myositis / myopathy by inclusion bodies, cataracts, retinitis pigmentosa with rhodopsin mutations,Frontotemporal lobar degeneration (FTLD), dementia, cadasil syndrome, hereditary cerebral hemorrhage with 10 amyloidosis, Alexander's disease, sepianopathies, familial amyloidic neuropathy, senile systemic amyloidosis, serpinopathies, AL (light chain) amyloidosis (primary systemic amyloidosis), amyloidosis AH (heavy chain), AA amyloidosis (secondary), aortic amyloidosis, ApoAI amyloidosis, ApoAII amyloidosis, ApoAIV amyloidosis, familial Finnish amyloidosis (FAF), lysozyme amyloidosis, fibrinogen amyloidosis, dialysis amyloidosis, myositis / myopathy by inclusion bodies, cataracts, retinitis pigmentosa with rhodopsin mutations,Frontotemporal lobar degeneration (FTLD), dementia, cadasil syndrome, hereditary cerebral hemorrhage with 10 amyloidosis, Alexander's disease, sepianopathies, familial amyloidic neuropathy, senile systemic amyloidosis, serpinopathies, AL (light chain) amyloidosis (primary systemic amyloidosis), amyloidosis AH (heavy chain), AA amyloidosis (secondary), aortic amyloidosis, ApoAI amyloidosis, ApoAII amyloidosis, ApoAIV amyloidosis, familial Finnish amyloidosis (FAF), lysozyme amyloidosis, fibrinogen amyloidosis, dialysis amyloidosis, myositis / myopathy by inclusion bodies, cataracts, retinitis pigmentosa with rhodopsin mutations,Senile systemic amyloidosis, serpinopathies, AL amyloidosis (light chain) (primary systemic amyloidosis), amyloidosis AH (heavy chain), AA amyloidosis (secondary), aortic amyloidosis, ApoAI amyloidosis, ApoAII amyloidosis, ApoAIV amyloidosis, familial amyloidosis of type Finnish (FAF), amyloidosis by lysozyme, amyloidosis by fibrinogen, amyloidosis by dialysis, myositis / myopathy by inclusion bodies, cataracts, retinitis pigmentosa with rhodopsin mutations,Senile systemic amyloidosis, serpinopathies, AL amyloidosis (light chain) (primary systemic amyloidosis), amyloidosis AH (heavy chain), AA amyloidosis (secondary), aortic amyloidosis, ApoAI amyloidosis, ApoAII amyloidosis, ApoAIV amyloidosis, familial amyloidosis of type Finnish (FAF), amyloidosis by lysozyme, amyloidosis by fibrinogen, amyloidosis by dialysis, myositis / myopathy by inclusion bodies, cataracts, retinitis pigmentosa with rhodopsin mutations,Amyloidosis by dialysis, myositis / myopathy by inclusion bodies, cataracts, retinitis pigmentosa with rhodopsin mutations,Amyloidosis by dialysis, myositis / myopathy by inclusion bodies, cataracts, retinitis pigmentosa with rhodopsin mutations,

15 medullary thyroid carcinoma, cardiac atrial amyloidosis, pituitary prolactinoma, hereditary reticular corneal dystrophy, lichen cutaneous amyloidosis, Mallory bodies. Amyloidosis due to corneal lactoferrin, pulmonary alveolar proteinosis, odontogenic amyloid tumor (Pindborg), cystic fibrosis, sickle cell disease or critical patient myopathy (MEC)

Other medical uses include treatment of bone-related disorders, such as osteoporosis or

20 osteoarthritis, etc., in which increased bone formation and reduction of bone resorption could be beneficial

Detailed description of the invention

Definitions

The amino acid sequences of the present invention contain one letter and three letter codes

25 conventional for naturally occurring amino acids, in addition to three-letter codes generally accepted for other amino acids, such as Aib (α-aminoisobuliric acid), Om (omitin), Dab (2,4-diamino-amino acid), Dap (2,3-acid) -diaminopropionic), Nle (norleucine), GABA (y-aminobuliric acid) or Ahx (acid (

aminohexanoic).

In addition, the following codes were used for the amino acids shown in Table 4 ·

30 Table 4:

<dl><dt>structure </dt><dd>first name code </dd></dl>

<dl><dt>H, '~ ,. OYOH quot ;,</dt><dd /></dl>

<dl><dt>(S) MeLys </dt><dd>(S) -a-methyl-lysine </dd><dt>(S) MeLys </dt><dd /></dl>

<dl><dt>quot; '</dt><dd>or ~ YOH HH, </dd></dl>

<dl><dt>(R) MeLys </dt><dd>(R) -a-methyl-lysine </dd><dt>(R) MeLys </dt><dd /></dl>

<dl><dt>or H. N ~ OH </dt><dd /></dl>

<dl><dt>quot; ', </dt><dd /></dl>

<dl><dt>(S) MeOm </dt><dd>(S) -a-methyl-omitine </dd><dt>(S) MeOm </dt><dd /></dl>

<dl><dt>structure </dt><dd>first name code </dd></dl>

<dl><dt>HNC} t OH NH, Pip </dt><dd>4-amino-piperidine-4-carboxylic acid Pip </dd></dl>

The term "exendin-4 native" refers to native exendin-4 having the sequence HGEGTFTSDLSKQMEEEAVRLFIEWLKNGGPSSGAPPPS-NH2 (SEQ ID NO: 1)

The invention provides peptide compounds as defined above.

The peptide compounds of the present invention comprise a linear backbone of aminocarboxylic acids linked by peptide, ie, carboxamide bonds. Preferably, the aminocarboxylic acids are aminocarboxylic acids and more preferably La-aminocarboxylic acids, unless otherwise indicated. The peptide compounds comprise a skeletal sequence of 39-40 aminocarboxylic acids

The peptide compounds of the present invention may have unmodified side chains, but carry at least one modification in one of the side chains.

For the avoidance of doubt, in the definitions provided herein it is generally intended that the sequence of the peptide moiety (11) be differentiated from the native exendin-4 at least in one of those positions that are established that allow variation. The amino acids within the peptide moiety (11) can be considered to be consecutively numbered from 0 to 40 in the conventional direction from the N-terminus to the C-terminus. Therefore, a reference to a "position" within the peptide moiety (11) should be created. , since it must refer to positions within native exendin-4 and other molecules, for example, in exendin-4, His is in position 1, Gly in position 2, Met in position 14,. And Being in position 39

The amino acid resins at position 14 having a lamellar chain with a -NH2 group, for example Lys, Orn, Dab or Dap, are conjugated to a functional group, for example acyl groups. Thus, one or more amino acids selected from the peptides in the present invention can carry a covalent linkage in their side chains. In some cases, those junctions can be lipophilic. These lipophilic side chain junctions have the potential to reduce the in vivo elimination of the peptides thus increasing their semividas in vivo.

The lipophilic linkage may consist of a lipophilic moiety which may be a branched or unbranched acyclic moiety, aliphatic

or unsaturated, and / or a cyclic moiety selected from one or more aliphatic or unsaturated homocycles or heterocycles, homocycles or aromatic heterocycles condensed or non-condensed, ether bonds, unsaturated bonds and substituents, for example hydroxy and / or carboxy groups. The lipophilic moiety can be linked to the peptide either by alkylation, reductive amination or by an amide bond, a carbamate bond or a sulfonamide in the case of amino acids bearing an amino group on its side chain

Non-limiting examples of lipophilic moieties that can be attached to the side chains of amino acids include fatty acids, for example C8-3 fatty acids (I such as palmitic acid, myristic acid, stearic acid and oleic acid, and / or cyclic groups as described above or derivatives thereof

There could be one or several connectors between the amino acid of the peptide and the lipophilic binding. Non-limiting examples of those connectors are 3-alanine, V-glutamic acid, o-glutamic acid, v-aminobutyric acid and / or [aminohexanoic acid or dipeptides, such as 3-Ala-3-Ala (also abbreviated 3A-3A herein) and / or V-Glu-V-Glu (also abbreviated VE-VE herein) in all its forms stereoisomer (S and R enantiomers)

Thus, a non-limiting example of a side chain linkage is palmitic acid which is covalently bound to the α-amino group of glutamic acid forming an amide bond. The v-carboxy group of this substituted glutamic acid can form an amide bond with the side chain amino group of a lysine within the peptide

In another aspect, the present invention provides a composition comprising a compound of the invention as described herein, or a solvate salt thereof, in admixture with a carrier.

The invention also provides the use of a compound of the present invention for use as a medicament, particularly for the treatment of a condition as described below.

The invention also provides a composition in which the composition is a pharmaceutically acceptable composition, and the carrier is a pharmaceutically acceptable carrier.

Synthesis of peptides

The skilled person knows a variety of different methods for preparing peptides that are described in the present invention. These methods include, but are not limited to. synthetic approaches and recombinant gene expression. Thus, a form of preparation of these peptides is the synthesis in solution or on a solid support and subsequent isolation and purification. A different form of preparation of the peptides is gene expression in a host cell into which a DNA sequence encoding the peptide has been introduced. Alternatively, gene expression can be achieved without using a cell system. The methods described above can also be combined in any way

A preferred way to prepare the peptides of the present invention is the solid phase synthesis on a suitable resin. Peptide solid phase synthesis is a well-established methodology (see, for example, Stewart and Young, Solid Phase Peptide Synthesis, Pierce Chemical CO., Rockford, IU, 1984, E. Atherton and RC Sheppard, Solid Phase Peptide Synthesis. A Practical Approach, Oxford-IRL Press, New York, 1989). Solid phase synthesis is initiated by attaching an amino acid protected from the N-terminus with its carboxy terminus to an inert solid support carrying a cleavable linker. This solid support can be any polymer that allows coupling of the initial amino acid, for example, a trityl resin, a chlorotryril resin, a Wang resin or a Rink resin in which the bond of the carboxy group (or carboxamide for the Rink resin) to the resin is acid sensitive (if Fmoc strategy is used). The polymer support must be stable in the conditions used to deprotect the Q-amino group during peptide synthesis.

After coupling the first amino acid to the solid support, the a-amino protecting group of this amino acid is removed. The remaining protected amino acids are then coupled one after the other in the order represented by the peptide sequence using appropriate amide coupling reagents, for example, BOP, HBTU, HATU or DIC (N, N'-diisopropylcarbodiimide) f HOBt (1-hydroxybenzotriazole), in which BOP, HBTU and HATU are used with tertiary amine bases. Alternatively, the released N-terminus can be functionalized with different groups of amino acids, for example, carboxylic acids, etc.

Normally, the reactive groups on the side chain of the amino acids are protected with suitable blocking groups. These protecting groups are removed after the desired peptides have been assembled. They are eliminated concomitantly with the cleavage of the desired product from the resin under the same conditions. Protective groups and methods for introducing protecting groups can be found in Protective Groups in Organic Synthesis, 38 ed., Greene, T. W. Wuts, PGM, Wiley &amp; Sons (New York: 1999)

In some cases it may be desired to have side chain protecting groups that can be selectively removed while other side chain protecting groups remain intact. In this case, the released functionality can be selectively functionalized. For example, a lysine can be protected with a protective group ivDde ([1- (4,4-dimethyl-2,6-dioxocyclohex-1-ylidene) -3-methylbutyl) (SR Chhabra et al., Tetrahedron Let. , (1998), 1603) which is labile to a very nUcleophilic base, for example 4% hydrazine in DMF (dimethylformamide). Thus, if the N-terminal amino group and all side chain functionalities are protected with acid-labile protecting groups, the ivDde group can be selectively removed using 4% hydrazine in DMF and the corresponding free amino group can then be further modified, for example by acylation. The lysine can alternatively be coupled to a protected amino acid and the amino group of this amino acid can then be deprotected resulting in another free amino group that can be acylated or bound to additional amino acids

Finally, the peptide is cleaved from the resin. This can be achieved using King's mix (DS King, C_ G. Fields,

G. 8. Fields, Inl. J. Peptide Protein Res. 36, 1990, 255-266). The starting material can then be purified by chromatography, for example, preparative RP-HPLC, if necessary.

Power

As used herein, the term "potentia"; or "in vitro potency"; is a measure of the ability of a compound to activate receptors for GLP-1, GIP or glucagon in a cell-based assay. Numerically expressed as the "CESO value", which is the effective concentration of a compound that induces an increase. at 50% response (e.g., intracellular cAMP formation) in a dose-response experiment

Therapeutic uses

The compounds of the invention are agonists for the GLP-1 and the GIP receptor receptors, in addition to the glucagon receptor optionally (eg "dual or trigonal agonists"). Such peptides which are coagonists of GIPfGLP-1, or tri-agonists of GIP / GLP-1 / glucagon, may provide therapeutic benefit to address a clinical need to target metabolic syndrome allowing the simultaneous treatment of diabetes and obesity.

Metabolic syndrome is a combination of medical conditions that, when produced together, increase the risk of developing type 2 diabetes, in addition to atherosclerotic vascular disease, for example, heart disease and stroke. The definition of medical parameters for the metabolic syndrome includes diabetes mellitus, glucose intolerance, high fasting glucose, insulin resistance, urinary albumin secretion, central obesity, hypertension, high triglycerides, high LDL cholesterol and reduced cholesterol HOL.

Obesity is a medical condition in which excess body fat has accumulated to such an extent that it can have an adverse effect on health and life expectancy and because of its increasing prevalence in adults and children it has become one of the main avoidable causes of death in the modern world. Increases the likelihood of various other diseases, which include heart disease, type 2 diabetes, obstructive sleep apnea, certain types of cancer, in addition to osteoarthritis, and most commonly occurs by a combination of excessive food intake, energy expenditure reduced, in addition to genetic susceptibility.

Diabetes mellitus, often simply called diabetes, is a group of metabolic diseases in which a person has high blood sugar levels, both because the body does not produce enough insulin, and because the cells do not respond to insulin that is produced. The most common types of diabetes are: (1) type 1 diabetes, in which the body fails to produce insulin; (2) type 2 diabetes, in which the body fails to use insulin properly, combined with an increase in insulin deficiency over time, and (3) gestational diabetes, in which women develop diabetes due to their pregnancy. All forms of diabetes increase the risk of long-term complications, which usually develop after many years. Most of these long-term complications are based on damage to the blood vessels and can be divided into the two categories "macrovascular" disease, which occurs from atherosclerosis of larger blood vessels, and "microvascular" disease, which It occurs from damage of small blood vessels. Examples of macrovascular disease conditions are ischemic heart disease, myocardial infarction, cerebrovascular accident and peripheral vascular disease. Examples of microvascular diseases are diabetic retinopathy, diabetic nephropathy, as well as diabetic neuropathy. which occurs from damage of small blood vessels. Examples of macrovascular disease conditions are ischemic heart disease, myocardial infarction, cerebrovascular accident and peripheral vascular disease. Examples of microvascular diseases are diabetic retinopathy, diabetic nephropathy, as well as diabetic neuropathy. which occurs from damage of small blood vessels. Examples of macrovascular disease conditions are ischemic heart disease, myocardial infarction, cerebrovascular accident and peripheral vascular disease. Examples of microvascular diseases are diabetic retinopathy, diabetic nephropathy, as well as diabetic neuropathy.

The receptors for GLP-1 and GIP, in addition to glucagon, are members of the transmembrane-spanning heterotrimeric protein G-coupled receptor family. They are structurally related to each other and share not only a significant level of sequence identity, but also have similar mechanisms of recognition of ligands and intracellular signaling pathways

Similarly, the GLP-1, GIP and glucagon peptides share regions of high identity / sequence similarity. GLP-1 and glucagon are produced from a common precursor, preproglucagon, which is differentially processed in a specific tissue-to-give manner. for example. GLP · 1 in intestinal endocrine cells and glucagon in pancreatic islet alpha cells. GIP is derived from a precursor of the proGRO major proGIP and is synthesized and released from K cells located in the small intestine

The incretin peptide hormones GLP-1 and GIP are secreted by intestinal endocrine cells in response to food and account for up to 70% of insulin secretion stimulated by food. Evidence suggests that GLP-1 secretion is reduced in subjects with impaired glucose tolerance or type 2 diabetes, while sensitivity to GLP-1 is still preserved in these patients. Thus, the targeting of the GLP-1 receptor with suitable agonists offers an attractive approach for the treatment of metabolic disorders, including diabetes. The receptor for GLP-1 is widely distributed, being found mainly in pancreatic islets, brain, heart, kidney and gastrointestinal tract. In the pancreas, GLP-1 acts in a manner strictly dependent on glucose, increasing the secretion of insulin from beta cells. This glucose dependence shows that activation of GLP · 1 receptors is unlikely to produce hypoglycemia. The GIP receptor is also widely expressed in peripheral tissues including pancreatic islets, adipose tissue, stomach, small intestine, heart, bone, lung. , kidney, testes, adrenal cortex, pituitary, endothelial cells, trachea, spleen, thymus, thyroid and brain. According to its biological function as incretin hormone, pancreatic B cells express the highest levels of the receptor for GIP in humans. There is some clinical evidence that signaling mediated by the GIP receptor could be altered in patients with T20M, but it is shown that the action of GIP is reversible and could be restored with improvement of the diabetic state.

At the level of bela cells, it has been shown that GLP-1 and GIP promote glucose sensitivity, neogenesis, proliferation, transcription of proinsulin and hypertrophy, as well as antiapoptosis. It could be anticipated that a peptide with dual agonist activity for the GLP-1 and GIP receptor has an additive or synergistic antidiabetic benefit. Other relevant effects of GLP · 1 beyond the pancreas include delayed gastric emptying, high satiety, decreased food intake, reduced body weight, in addition to neuroprotective and cardioprotective effects. In patients with type 2 diabetes, such extrapancreatic effects may be particularly important, considering the high rates of comorbidities such as obesity and cardiovascular disease. Other GIP actions in peripheral tissues beyond the pancreas include increased bone formation and decreased bone resorption, as well as neuroprotective effects that could be beneficial for the treatment of osteoporosis and cognitive defects such as Alzheimer's disease. Glucagon is a 29 amino acid peplidic hormone that is produced by pancreatic alpha cells and released into the bloodstream when glucose in circulation is low. An important physiological function of glucagon is to stimulate glucose output in the liver, which is a process that provides the main counterregulatory mechanism for insulin in the maintenance of glucose homeostasis in vivo. in addition to neuroprotective effects that could be beneficial for the treatment of osteoporosis and cognitive defects such as Alzheimer's disease. Glucagon is a 29-amino acid peplidic hormone that is produced by pancreatic alpha cells and released into the bloodstream when glucose in circulation is low . An important physiological function of glucagon is to stimulate glucose output in the liver, which is a process that provides the main counterregulatory mechanism for insulin in the maintenance of glucose homeostasis in vivo. in addition to neuroprotective effects that could be beneficial for the treatment of osteoporosis and cognitive defects such as Alzheimer's disease. Glucagon is a 29-amino acid peplidic hormone that is produced by pancreatic alpha cells and released into the bloodstream when glucose in circulation is low . An important physiological function of glucagon is to stimulate glucose output in the liver, which is a process that provides the main counterregulatory mechanism for insulin in the maintenance of glucose homeostasis in vivo.

Glucagon receptors are also expressed, however, in extrahepatic tissues such as the kidney, heart, adipocytes, lymphoblasts, brain, retina, adrenal gland and gastrointestinal tract, suggesting a wider physiological function beyond the homeostasis of glucose . Consequently, recent studies have reported that glucagon has therapeutic positive effects on energy management, which includes the stimulation of energy expenditure and thermogenesis, accompanied by the reduction of food consumption and the loss of body weight. Taken together, the stimulation of glucagon receptors could be useful in the treatment of obesity and metabolic syndrome

Oxyintomodulin is a peptide hormone consisting of glucagon with an extension of the C-terminus that encompasses eight amino acids. Like GLP-1 and glucagon, it is preformed into preproglucagon and cleaved and secreted in a tissue-specific fashion by endocrine cells of the small intestine. Oxintomodulin is known to stimulate both GLP-1 and glucagon receptors and is therefore the prototype of a dual agonist

As GLP-1 and GIP are known for their antidiabetic effects, GLP-1 and glucagon are both known for their suppressive effects of food consumption and glucagon is also a mediator of additional energy expenditure, it is conceivable that a combination of activities of the two hormones in a molecule can give a powerful medication for the treatment of metabolic syndrome and in particular its components diabetes and obesity.

Accordingly, the compounds of the invention can be used for the treatment of glucose intolerance, insulin resistance, pre-diabetes, high fasting glucose, type 2 diabetes, hypertension, dyslipidemia, arteriosclerosis, coronary heart disease, peripheral arteries, stroke or any combination of these individual disease components

In addition, they can be used to control appetite, food and calorie intake, increase in energy expenditure, prevention of weight gain, promotion of weight loss, reduction of excess body weight and overall obesity treatment, which includes morbid obesity

Other disease states and health conditions that could be treated with the compounds of the invention are obesity-linked amation, gallbladder disease linked to obesity and obesity-induced sleep apnea.

Although all of these conditions could be associated directly or indirectly with obesity, the effects of the compounds of the invention can be measured in whole or in part by an effect on body weight,

or independent of it.

In addition, diseases to be treated are osteoporosis and neurodegenerative diseases such as Alzheimer's disease or Parkinson's disease, or other degenerative diseases as described above.

In comparison with GLP-1, glucagon and oxintomodulin, exendin-4 has beneficial physicochemical properties, such as solubility and stability in solution and under physiological conditions (including enzymatic stability towards degradation by enzymes, such as DPP-4 or NEPl, which Thus, exendin-4 could serve as a good starting scaffold to obtain exendin-4 analogs with dual or even triple pharmacologies, for example, GLP-1-glucagon agonism and optionally in addition to glucagon

However, it has also been shown that exendin-4 is chemically labile due to the oxidation of methionine at position 14, in addition to the deamidation and isomerization of asparagine at position 28. Therefore, the stability could be further improved by substitution of methionine at position 14 and avoiding sequences known to be prone to degradation by the formation of aspartimide, especially Asp-Gly or Asn-Gly at positions 28 and 29.

Pharmaceutical compositions

The term "pharmaceutical composition" indicates a mixture that contains components that are compatible when mixed and that can be administered. A pharmaceutical composition may include one or more medicinal drugs. Additionally, the pharmaceutical composition may include vehicles, buffers, acidifying agents, alkalizing agents, solvents, adjuvants, tonicity adjuster, emollients, expanders, preservatives, physical and chemical stabilizers, for example, surfactants, antioxidants and other components, whether they are considered as active or inactive. Guidance for the skilled in the preparation of pharmaceutical compositions can be found, for example, in Remington: The Science and Practice of Pharmacy (20th ed.l ed. AR Gennaro A. R, 2000, Lippencott Williams &amp; Wilkins and in R C. Rowe et al.

The exendin-4 peptide derivatives of the present invention, or salts thereof, are administered in conjunction with a pharmaceutically acceptable carrier, diluent or excipient as part of a pharmaceutical composition. A "pharmaceutically acceptable vehicle" it is a vehicle that is physiologically acceptable (eg, physiologically acceptable pH), while retaining the therapeutic properties of the substance with which it is administered. Conventional acceptable pharmaceutical carriers and their formulations are known to one skilled in the art and are described, for example, in Remington: The Science and Practice of Pharmacy (20th ed.) Ed. A. R Gennaro A. R, 2000, LippencolI Williams &amp; Wilkins and in R C. Rowe et al. (Ed), Handbook of Pharmaceutical Excipients, PhP, May 2013 update.

In one embodiment, the carriers are selected from the group of buffers (e.g., citrate citric acid), acidifying (e.g., hydrochloric acid), alkalizing (e.g., sodium hydroxide), preservatives (e.g., phenol), co-solvents (e.g. for example, polyethylene glycol 400), tonicity adjusters (e.g., mannitol), stabilizers (e.g., surfactant, antioxidants, amino acids).

The concentrations used are in a range that is physiologically acceptable.

Acceptable pharmaceutical carriers or diluents include those used in formulations suitable for oral, rectal, nasal or parenteral administration (including subcutaneous, intramuscular, intravenous, intradermal and transdermal). The compounds of the present invention will normally be administered parenterally

The term "pharmaceutically acceptable salt" means salts of the compounds of the invention which are safe and effective for use in mammals. The pharmaceutically acceptable salts may include, but are not limited to, acid addition salts and basic salts. Examples of acid addition salts include chloride, sulfate, hydrogen sulfate, (hydrogen) phosphate, acetate, citrate, tosylate or mesylate salts. Examples of basic salts include salts with inorganic cations, for example, alkali metal or alkaline earth metal salts such as sodium, potassium, magnesium or calcium salts, and salts with organic cations such as amine salts. Other examples of pharmaceutically acceptable salts are described in Remington: The Science and Practice of Pharmacy, (20th ed.) Ed. A. R Gennaro A. R, 2000, Lippencott Williams &amp;

The term "solvate" means complexes of the compounds of the invention or salts thereof with solvent molecules, for example, organic solvent molecules and / or water.

In the pharmaceutical composition, the exendin-4 derivative may be in monomeric or oligomeric form.

The term "therapeutically effective amount" of a compound refers to a non-toxic, but sufficient amount of the compound, to provide the desired effect. The amount of a compound of formula necessary to achieve the desired biological effect depends on several factors, for example, the specific compound chosen, the intended use, the mode of administration and the clinical condition of the patient. An "effective" amount appropriate in any individual case may be determined by a routine experimentation in the art using routine experimentation. For example, the "therapeutically effective amount" of a compound of formula (1) is about 0.01 to 50 mg / dose, preferably 0.1 to 10 mg / dose.

The pharmaceutical compositions of the invention are those suitable for parenteral (for example, subcutaneous, intramuscular, intradermal or intravenous), oral, rectal, topical and oral administration (for example, sublingual), although the most appropriate mode of administration depends in each case individual of the nature and severity of the condition to be treated and of the nature of the compound of formula I used in each case.

Suitable pharmaceutical compositions may be in the form of separate units, for example, capsules, tablets and powders in vials or ampoules, each containing a defined amount of the compound; as powders or granules; as a solution or suspension in an aqueous or non-aqueous liquid; or as an oil in water or water in oil emulsion. It can be provided in single or multiple dose injectable form, for example, in the form of a pen. The compositions can be prepared, as already mentioned, by any suitable pharmaceutical method which includes a step in which the active principle and the vehicle (which can consist of one

or more additional components) are put in contact

In certain embodiments, the pharmaceutical composition may be provided together with a device for the application, for example, together with a syringe, an injection pen or an auto-injector. Such devices can be provided separately from a pharmaceutical composition or pre-loaded with the pharmaceutical composition

Combination therapy

The compounds of the present invention, dual agonists for GLP-1 and GIP receptors, trigonal agonists for the GLP-1, GIP and glucagon receptors, can be combined widely with other pharmacologically active compounds, such as all drugs mentioned in the Rote Liste 2012 and / or the Rote Liste 2013, for example, with all the antidiabetics mentioned in the Rote Liste 2012, chapter 12, and / or the Rote Liste 2013, chapter 12, all the weight-reducing agents or suppressors of the appetite mentioned in Rote Liste 2012, chapter 1, and / or Rote Liste 2013, chapter 1, all lipid-lowering agents mentioned in Rote Liste 2012, chapter 58, and / or Rote Liste 2013, chapter 58, all antihypertensives and nephroprotectors mentioned in the Rote Liste 2012, and / or the Rote Liste 2013,or all diuretics mentioned in Rote Liste 2012, chapter 36, and / or Rote Liste 2013, chapter 36.

The active ingredient combinations can be used especially for a synergistic improvement in the action. They can be applied either by separate administration of the active ingredients to the patient or in the form of combination products in which a plurality of active ingredients is present in a pharmaceutical preparation. When the active ingredients are administered by separate administration of the active ingredients, this can be done simultaneously or successively.

Most of the active ingredients mentioned hereinafter are disclosed in the USP Oictionary of USAN and International Orug Names, US Pharmacopeia, Rockville 2011

Other active substances which are suitable for such combinations include in particular those which, for example, enhance the therapeutic effect of one or more active substances with respect to one of the aforementioned indications and / or which make it possible to reduce the dosage of one or more active substances .

Therapeutic agents that are suitable for combinations include, for example, antidiabetic agents such as:

Insulin and insulin derivatives, for example: Glargine / Lantus®, 270-330 U / ml insulin glargine (EP 2387989 A), 300 U / ml insulin glargine (EP 2387989 A), glulisine / Apidra®, getemir / Levemir®, lispro / Humalog® / Liprolog®, degludec / DegludecPlus, aspart, basal insulin and analogs (eg, LY-2605541, LY2963016, NN1436), PEGylated insulin lispro, Humulin®, Unjeta, SuliXen®, NN1045, insulin more Symlin, PE0139, fast-acting and slow-acting insulins (eg, Linjeta, PH20, NN1218, HinsBet), ~ APC-002) hydrogel, oral, inhaled, transdermal and sublingual insulins (eg Exubera®, Nasulin, Afrezza, Tregopil, TPM 02, Capsulin, Oral-Iyn®, oral insulin Cobalamin®, ORMO0801, NN1953, NN1954, NN1956, VIAtab, oral insulin Oshadi).Additionally, those insulin derivatives that are bound to albumin or another protein by a bifunctional linker are also included.

GLP-1, GLP-1 amylogues and GLP-1 receptor agonists, for example: lixisenatide {AVE0010 I ZP10 I Lyxum ia, exenatide {exend ina-4 / Byeture {Bydureon {ITCA 650 {AC-2993, liraglutida { Victoza, semaglutide, taspoglutide, Syncria I albiglutide, dulaglutide, rExendina-4, CJC-1134-PC, PB-1023, TTP..Q54, langlenatide I HM-11260C, CM-3, GLP-1 Choose, ORMO-0901, NN-9924, NN-9926, NN-9927, Nodexen, Viador-GLP-1, CVX-096, ZYOG-1, ZYO-1, GSK-2374697, DA-3091, MAR-701, MAR709, ZP-2929, ZP-3022, TT-401, BHM..Q34, MOO-6030, CAM-2036, DA15864, ARI-2651, ARI-2255, exenatide-XTEN and glucagon-Xten

OPP-4 inhibitors, for example: alogliptin {Nesin, Trajenta / linagliptin / BI-1356 / Ondero / Trajenta I Tradjenta I Trayenta I Tradzenta, saxagliptin I Onglyza, sitagliptin I Januvia {Xelevia I Tesave {Janumet {Velmetia, Galvus {vildagliptin , anagliptin, gemigliptin, teneligliptin, melogliptin, trelagliptin, OA-1229, omarigliptin I MK-3102, KM-223, evogliptin, ARI-2243, PBL-1427, pinoxacin.

Inhibitors of SGL T2, for example: Invokana I canaglifozina, Forxiga {dapagliflozina, remoglifozina, sergliflozina, empagliflozina, ipragliflozina, tofogliflozina, luseogliflozina, LX-4211, ertuglifozina {PF-04971729, R0-4998452, EGT0001442, KGA-3235 / OSP- 3235, lIK066, SBM-TFC..Q39,

Biguanides (for example, metformin, buformin, phenformin), thiazolidinediones (e.g., pioglitazone, rivoglitazone, rosiglitazone, troglitazone), dual PPAR agonists (e.g., aleglitazar, murag1ylazar, tesaglitazar), sulfonylureas (e.g., tolbutamide, glibenclamide, glimepiride / amaryl, glipizide), meglitinides (eg, nateglinide, repaglinide, mitiglinide), alpha-glucosidase inhibitors (eg, acarbose, miglitol, voglibose), amylin and amylin analogues (eg, pramlintide, Symlin).

GPR119 agonists (eg, GSK-263A, PSN-821, MBX-2982, APD-597, ZYG-19, OS-8500), GPR40 agonists (eg, fasiglifam (TAK-875, TUG-424, P -1736, JTT-851, GW9508).

Other suitable combination components are: Cycloset, 11-beta-HSD inhibitors (for example, LY2523199, BMS770767, RG-4929, BMS816336, AZO-8329, HSO-016, BI-135585), glucokinase activators (e.g. TTP-399, AMG-151, TAK-329, GKM-001), DGAT inhibitors (for example, LCO-908), protein tyrosine phosphatase 1 inhibitors (for example, trodusquemin), glucose-6-phosphatase inhibitors , inhibitors of fructose-1,6-bisphosphatase, inhibitors of glycogen phosphonlase, inhibitors of phosphoenol pyruvate carboxykinase,

S

glycogen synthase kinase inhibitors, pyruvate dehydrokinase inhibitors, alpha2 antagonists, CCR-2 antagonists, SGLT-1 inhibitors (eg, LX-2761).

Also suitable are one or more lipid-lowering agents as combination components, such as for example: inhibitors of HMG-CoA reductase (eg, simvastatin, atorvastatin), fibrates (eg, bezafibrate, fenofibrate), nicolinic acid and the derivatives of the same (for example, niacin), PPAR- agonists (alpha, gamma or alpha / gamma) or modulators (for example, aleglitazar), PPAR-delta agonists, ACAT inhibitors (for example, avasimibe), absorption inhibitors of cholesterol (e.g., ezetimibe), bile acid binding substances (e.g., cholestyramine), inhibitors of ileal bile acid transport, MTP inhibitors or PCSK9 modulators.

Compounds that elevate HOL such as: CETP inhibitors (eg, torcetrapib, anacetrapid, dalcetrapid, evacetrapid, JTT-302, DRL-17822, TA-899S) or ABC1 regulators.

Other suitable combination components are one or more active substances for the treatment of obesity, such as, for example · sibutramine, tesofensin, orthothamid, cannabinoid-1 receptor antagonists, MCH-1 receptor antagonists, MC4 receptor agonists , NPYS2 or NPY2 antagonists (eg, velneperit), beta-3-agonists, leptin or leptin mimetics, 5HT2c receptor agonists (eg, lorcaserin), or combinations of bupropionane-1-trexone, bupropionazzonisamide, bupropionalphentermine, or pramlintide / metreleptin

Other suitable combination components are:

Other gastrointestinal peptides such as peptide YY 3-36 (PYY3-36) or analogs thereof, pancreatic polypeptide (PP) or analogs thereof

Agonists or antagonists of glucagon receptors, agonists or antagonists of GIP receptors, antagonists or inverse agonists of ghrelin, xenin and analogs thereof

In addition, combinations with drugs are suitable for influencing arterial hypertension, chronic heart failure or atherosclerosis such as, for example: angiotensin II receptor antagonists (for example, lelmisartan, candesartan, valsarlan, losartan, eprosartan, imesarlan, olmesartan, lasosartan , azilsarlan), ACE inhibitors, ECE inhibitors, diuretics, beta-blockers, calcium antagonists, centrally acting hypertensives, alpha-2-adrenergic receptor antagonists, neutral endopeptidase inhibitors, thrombocyte aggregation inhibitors and others or combinations thereof.

In another aspect, the present invention relates to the use of a compound according to the invention, or a physiologically acceptable salt thereof, combined with at least one of the active substances described above as a combination component, for preparing a medicament that is suitable for the treatment or prevention of diseases or conditions that can be affected by binding to GLP-1 and glucagon receptors and modulating their activity. This is preferably a disease in the context of the metabolic syndrome, particularly one of the diseases or conditions listed above, most particularly diabetes or obesity or complications thereof.

The use of the compounds according to the invention, or a physiologically acceptable salt thereof, in combination with one or more active substances can take place simul- taneously, separately or sequentially

The use of the compound according to the invention, or a physiologically acceptable salt thereof, in combination with another active substance, can take place simultaneously or at staggered times, but particularly within a short period of time. If administered simultaneously, the two active substances are administered to the patient together; if used at staggered times, the two active substances are administered to the patient within a period less than or equal to 12 hours, but particularly less than or equal to 6 hours.

Accordingly, in another aspect, the present invention relates to a medicament comprising a compound according to the invention or a physiologically acceptable salt of a compound such and at least one of the active substances described above as combination components, optionally together with one or more inert vehicles and or thinners

The compound according to the invention, or physiologically acceptable salt or solvate thereof, and the additional active substance to be combined therewith can both be present together in a formulation, for example, a tablet or capsule, or separately in two formulations identical or different, for example, as the so-called kit of parts.

LEGENDS OF THE FIGURES

Figure 1. Effect of the sc administration of the compound SEO ID NO: 11 to 3 ~ gfkg and 10 ~ gfkg on the body weight in female C57BU6NCrI mice with diet-induced obesity (DIO) after the chronic treatment of 3 weeks once at day. The data are average ± EEM

Figure 2. Effect of the sc administration of the compound SEO ID NO: 11 to 3 ~ glkg and 10 ~ glkg on the body weight in female C57BU6NCrI mice with diet-induced obesity (DIO) after the chronic treatment of 3 weeks once at day. Changes in body weight were calculated as the relative change from the initial level. The data are average t EEM

5 Figure 3. Effect of 4 weeks of treatment with SEO ID NO: 11 to 3 and 10 ~ glkg, sc on fasting glucose in diabetic dbdb mice, represented as the change from the initial level (O mmolll, dia -7) . The data are average t EEM.

Figure 4. Effect of 4 weeks of treatment with SEO ID NO: 11 to 3 and 10 ~ g / kg, sc on HbA1c in diabetic dbdb mice, represented as the change from the initial level (O%, day -7). The data are mean ± EEM.

10 Figure 5. Effect of 4 weeks of treatment with SEO ID NO: 11 to 3 and 10 ~ glkg, sc on oral glucose tolerance in diabetic dbdb mice, represented as the change from the initial level (t = 0 min, O mmol, immediately before the administration of glucose). The data are average ± EEM

Figure 6. Effect of 4 weeks of treatment with SEO ID NO: 11 to 3 and 10 ~ glkg, sc on oral glucose tolerance in diabetic dbdb mice, represented as the area under the glucose curve (ABC of

15 glucose). The data are average t EEM. Figure 7. Treatment effect with SEO ID NO: 11, SEO ID NO: 12 and SEO ID NO; 15 to 3 ~ glkg, sc on the reduction of glucose in female diabetic dbdb mice fasting, represented as the change from the initial level. The data are average t EEM

Figure 8. Effect of the sc administration of the compound SEO ID NO: 11 to 1, 10 and 100 ~ g / kg on gastric emptying and intestinal transit in female NMRI mice. The data are average t EEM. a) - + Gastric emptying b) ..... Transit of the small intestine with respect to the length of the small intestine METHODS The abbreviations used are the following: 25 AA amino acid cAMP cyclic adenosine monophosphate Boc tert-butyloxycarbonyl BOP hexafluorophosphate benzotriazol-1-yloxy) tris (dimethylamino) phosphonium BSA bovine serum albumin 30 IBu butyl tertiary Rio de 1- (4,4 -dimethyl-2,6-dioxocyclohexylidene) -ethyl IVOde 1- (4,4-d imethyl) -2, 6-dioxocyclohexylidene) 3-methyl-butyl Die N, N'-diisopropylcarbodiimide DIPEA N,

<dl><dt>HBTU </dt><dd>2- (1 H-benzotriazole.1-yl) -1, 1, 3,3-tetramethyl-uron hexafluorophosphate </dd></dl>

<dl><dt>HE PES </dt><dd>2- [4- (2-hydroxyethyl) piperazin-1-yl-methanesulfonic acid </dd></dl>

<dl><dt>HOBt </dt><dd>1-hydroxybenzotriazole </dd></dl>

<dl><dt>HOSu </dt><dd>N-hydroxysuccinimide </dd></dl>

HPLC high performance liquid chromatography

HTRF homogeneous fluorescence resolved over time

IBMX 3-isobutyl-1-methylx ntin a

CUEM liquid chromatography / mass spectrometry

Palm palmiloilo

PSS phosphate buffered saline

PEG polyethylene glycol

pe pharmacokinetics

RP-HPLC reverse phase high resolution liquid chromatography

Stea

TFA trifluoroacetic acid

Trityl Trt

uv ultraviolet

General synthesis of peptide compounds

Materials:

Different Rink amide resins (4- (2 ', 4'-dimethoxyphenyl-Fmoc-aminomethyl) -phenoxyacetamidonorleucilaminomethyl resin, Merck Biosciences resin; 4 - [(2,4-dimethoxyphenyl) (Fmoc-amino) methyljphenoxyacetamidomethyl resin, were used; Agilent Technologies) for the synthesis of peptide amides with fillers in the range of 0.3-0.4 mmollg.

Natural amino acids protected with Fmoc from Protein Technologies Inc., Senn Chemicals, Merck Biosciences, Novabiochem, Iris Biotech or Bachem were purchased. The following conventional amino acids were used in all the syntheses: Fmoc-L-Ala-OH, Fmoc-Arg (Pbf) -OH, Fmoc-L-Asn (Trt) -OH, Fmoc-L-Asp (OtBu) -OH, Fmoc-L-Cys (Trt) -OH, Fmoc-L-Gln (Trt) -OH, Fmoc-L-Glu {OtBu) -OH, Fmoc-Gly-OH, Fmoc-L-His (Trt) -OH, Fmoc-L-ne-OH, Fmoc-L-Leu-OH, Fmoc-L-Lys {Boc) -OH, Fmoc-L-Met-OH, Fmoc-L · Phe-OH, Fmoc-L-Pro-OH , Fmoc-L-Ser {tBu) -OH, Fmoc-L-Thr {tBu) OH, Fmoc-L-Trp {Boc) -OH, Fmoc-L-Tyr (tBu) -OH, Fmoc-L-Val- OH.

In addition, the following special amino acids were purchased from the same suppliers as before: Fmoc-LLys {ivDde) ​​-OH, Fmoc.L-Lys {Mmt) -OH, Fmoc-Aib-OH, Fmoc-D-Ser {tBu) - OH, Fmoc-D-Ala-OH, Boc-L-His {Boc) -OH (available as toluene solvates) and Boc-L-His (Trt) -OH

Solid phase peptide syntheses were performed, for example, on a Prelude peptide synthesizer (Protein Technologies Inc) or similar automated synthesizer using conventional Fmoc chemistry and activation with HBTUfOIPEA. DMF was used as solvent. Deprotection: 20% piperidinal DMF for 2 x 2.5 min. Washes: 7 x DMF. Coupling 2: 5: 10 AA 200 mM I HBTU 500 mM I DI PEA 2 M in DMF 2 x for 20 min Washes: 5 x DMF.

In cases where a side chain of Lys was modified, Fmoc-L-Lys (ivDde) ​​-OH or Fmoc-L-Lys (Mmt) -OH was used in the corresponding position. After completion of the synthesis, the ivDde group was removed according to a modified procedure from the literature (SR Chhabra et al., Tetrahedron Let !. 39, (1998), 1603), using 4% hydrated hydrazine in DMF. The Mmt group was removed by repeated treatment with 1% TFA in dichloromethane. The following acylations were snowed out by treating the resin with the N-hydroxysuccinimide esters of the desired acid or using coupling reagents such as HBTU / DIPEA or HOBVDIC.

All the peptides that had been synthesized were excised from the resin with the King cleavage mixture consisting of 82.5% TFA, 5% phenol, 5% water, 5% thioanisole, 2.5% EDT . The crude peptides were then precipitated in diethyl or diisopropyl ether, centrifuged and lyophilized. The peptides were analyzed by analytical HPLC and checked by ESI mass spectrometry. The crude peptides were purified by a conventional preparative HPLC purification procedure.

HPlC I analytical UPlC

Method A: Analytical UPLC I EM was performed in a Waters UPLC system with a C18 column of 1.7 11m

5 Waters UPLC HSS (2.1 x 100 mm) at 40 ° C with a gradient elution at a flow rate of 0.5 mlfmin and monitored at 215 and 280 nm. The gradients were established as 10% B to 90% B for 15 min and then 90% B for 1 min or as 15% BaSO% B for 12.5 min and then 50% B to 90% B for 3 min. Buffer A = 0.1% formic acid in water and B = 0.1% formic acid in acetonitrile.

A Waters LCT Premier time-of-flight instrument was used as a mass analyzer equipped with an electrospray in the positive ion mode.

Method B: Detection at 210-225 nm, optionally coupled to Waters Premier LCT mass analyzer, positive spray electro-ion mode

column · Waters ACQUITY UPLC® CSH TM C18 1.7 IJm (150 x 2.1 mm) at 50 oC

solvent · H20 + 0.5% TFA · ACN + 0.35% TFA (flow 0.5 ml / min)

gradient: 80:20 (O min) to 80:20 (3 min) to 25:75 (23 min) to 2:98 (23.5 min) to 2:98 (30, S min) to 80:20 ( 31 min) a

80:20 (37 min)

Method C: Detection at 215 nm

column · Aeris Peptide, 3.6 IJm, XB-C18 (250 x 4.6 mm) at 60 oC

solvent · H20 + 0.1% TFA: ACN + 0.1% TFA (flow 1.5 mlfmin)

gradient: 90:10 (O min) to 90:10 (3 min) to 10:90 (43 min) to 10:90 (48 min) to 90:10 (49 min) to 90:10 (50 min)

Method D: Detection at 214 nm

column: Waters X-Bridge C18 3.5 11m 2.1 x 150 mm

solvent · H20 + 0.5% TFA. ACN (flow 0,55 mlfmin)

gradient: 90:10 (O min) to 40:60 (5 min) to 1:99 (15 min)

Method E: Detection at 210-225 nm, optionally coupled to a Waters Premier LCT mass analyzer, positive spray electro-ion mode

column: Waters ACQUITY UPLCIIP BEHTM C18 1.7 IJm (150 x 2.1 mm) at 50 oC

solvent · H20 + 1% FA · ACN +1% FA (flow 0.9 mUmin)

gradient 95: 5 (O min) to 95: 5 (2 min) to 35:65 (3 min) to 65:35 (23.5 min) to 5:95 (24 min) to 95: 5 (26 min) to 95: 5 (30 min)

General preparative HPlC purification procedure:

The crude peptides were purified on both an Akta purification system and on a Jasco semiprep HPLC system. Preparative RP-C18-HPLC columns of different sizes and with different flow rates were used depending on the amount of crude peptide to be purified. They were used

25 acetonitrile + 0.05 0.1% TFA (8) and water + 0.05 to 0.1% TFA (A) as eluents. Alternatively, a buffer system consisting of acetonitrile and water with minor amounts of acetic acid was used. The fractions containing product were collected and lyophilized to obtain the purified product, usually as TFA or acetate salt

Solubility and stability test of exendin-4 derivatives

Before the test of solubility and stability of a batch of peptides, its content was determined. Therefore, two parameters were investigated, their purity (HPLC-UV) and the amount of batch salt load (ion chromatography)

For the solubility tests, the target concentration was 1.0 mg / ml of pure compound. Therefore, solutions of solid samples were prepared in different buffer systems with a concentration of 1.0 mg / ml of compound based on the previously determined content. UV-HPLC was performed after 2 h of gentle agitation of the supernatant, which was obtained by 20 min of centrifugation at 4000 rpm.

The solubility was then determined by comparing with the peak UV areas obtained with a stock solution of the peptide at a concentration of 2 mgfml in pure water or a variable amount of acetonitrile (optical control of which the compound was dissolved). This analysis also served as an initial point (tO) for the stability test.

For the stability test, an aliquot of the supernatant obtained for the solubility was stored for 7 days at 25 ° C. After this lapse of time, the sample was centrifuged for 20 min at 4000 rpm and the supernatant was analyzed with HPLC-UV.

For the determination of the amount of the remaining peptide the peak areas of the target compound were compared to 10 and t7, yielding the "remaining peptide%", following the equation

Remaining peptide% = [(peptide of peak area 17) x 100] / peptide of peak area tO.

The amount of soluble degradation products was calculated from the comparison of the sum of the peak areas of all observed impurities reduced by the sum of the peak areas observed at 10 (ie, to determine the amount of species related to peptides). recently formed). This value was given in percentage relation with respect to the initial amount of peptide in 10, following the equation:

% of soluble degradation products = {[(sum of peak areas of impurities 17) - (sum of peak areas of impurities 10)] x 100} / peak area peptide 10

The possible difference of the sum of the "remaining peptide%" and "% of soluble degradation products · up to 100% reflects the amount of peptide that did not remain soluble after stress conditions following the equation

% precipitate = 100 - ([% remaining peptide] + [% soluble degradation products])

This precipitate includes non-soluble degradation products, polymers and / or fibrils, which have been removed from the analysis by centrifugation

Chemical stability is expressed as "remaining peptide%".

Anion chromatography

Instrument: Dionex ICS-2000, prefolumn: Ion Pac AG-18 2 x 50 mm (Dionex) / AS18 2 x 250 mm (Dionex), eluent: aqueous sodium hydroxide, flow: 0.38 mlfmin, gradient: 0-6 min : 22 mM KOH, 6-12 min: 22-28 mM KOH, 1215 min: 28-50 mM KOH, 15-20 min: 22 mM KOH, suppressor: ASRS 300 2 mm, detection · conductivity

The method D or E has been used as HPLCfUPLC method

In vitro cell assays for the efficacy of the GIP receptor, GLP-1 receptor and the glucagon receptor

The agonism of compounds for the receptors was determined by functional assays measuring the cAMP response of HEK-293 cell lines stably expressing the GIP, GLP-1 or human glucagon receptor

The cAMP content of cells was determined using a Cisbio Corp. kit (No. of cal 62AM4PEC) based on HTRF (homogeneous fluorescence resolved over time). For the preparation, the cells were fractionated in T175 culture flasks and cultured overnight at near confluence in medium (DMEM f 10% FBS). The medium was then removed and the cells were washed with PBS lacking calcium and magnesium, followed by treatment with proteinase with Accutase (No. of lime from Sigma-Aldrich A6964). The detached cells were washed and resuspended in assay buffer (1 x HBSS, 20 mM HEPES, 0.1% BSA, 2 mM IBMX) and cell density was determined. They were then diluted to 400,000 cells / ml and 25 ~ I aliquots were dispensed into the wells of 96-well plates. For the measurement, 25 μl of test compound in assay buffer was added to the wells, followed by incubation for 30 minutes at room temperature. After the addition of HTRF reagents diluted in lysis buffer (kit components), the plates were incubated for 1 h, followed by measurement of the fluorescence ratio at 665 I 620 nm. The in vitro potency of agonists was quantified by determining the concentrations that caused 50% activation of the maximal response (EC50).

Bioanalytical screening method for the quantification of exendin-4 derivatives in mice and pigs

Mice were dosed with 1 mgfkg subcutaneously (mice were sacrificed and blood samples were collected after 0.25, 0.50, 1, 2, 4, B, 16 and 24 hours after the administration. analyzed after protein precipitation by mass spectrometry with liquid chromatography (EMfCL) .PK parameters and half-life were calculated using WinonLin Version 5.2.1 (non-compartmental model)

Gottlinger female minicerdose were dosed with 0.1 mgfkg subcutaneously (blood samples were collected after 0.25, 0.5, 1, 2, 4, 8, 24, 32, 48, 56 and 72 hours after the administration). Administration: Plasma samples were analyzed after protein precipitation by liquid chromatography-mass spectrometry (CLIEM) _ PK parameters and half-life were calculated using WinonLin Version 5_2_1 (non-compartmental model).

Gastric emptying and intestinal transit in mice

Female NMRI mice of a body weight between 20 and 30 g were used. The mice adapted to the housing conditions for at least one week

The mice fasted during the night, although they had water available all the time. On the day of the study, the mice were weighed, placed individually in cages and allowed access to 500 mg of feed for 30 min, while the water was removed. At the end of the 30-min feeding period, the remaining feed was removed and weighed. Then, the test compound f reference compound or its vehicle was administered in the control group subcutaneously_ 60 min later, to allow the compound to reach the relevant plasma exposure, a colored non-caloric bolus was instilled by nasogastric tube in the stomach_ After another 30 min, the animals were sacrificed and the stomach and small intestine were prepared. The full stomach was weighed, emptied, it was carefully cleaned and dried and weighed again. The content of the stomach, calculated as the weight of the full stomach subtracted from the weight of the empty stomach, indicated the degree of gastric emptying. The small intestine straightened without force and was measured in length. The distance from the gastric principle of the intestine to the stomach was then measured. tip of the bolus of intestinal content displaced further. The intestinal transit was facilitated as the ratio in percentage of this last distance and the total length of the small intestine. indicated the degree of gastric emptying_ The small intestine straightened without force and was measured in length_ Then the distance from the gastric principle of the intestine to the tip of the bolus of intestinal content moved further away was measured. The intestinal transit was facilitated as the ratio in percentage of this last distance and the total length of the small intestine. indicated the degree of gastric emptying_ The small intestine straightened without force and was measured in length_ Then the distance from the gastric principle of the intestine to the tip of the bolus of intestinal content moved further away was measured. The intestinal transit was facilitated as the ratio in percentage of this last distance and the total length of the small intestine.

Statistical analyzes were performed with Everstat 6.0 by unilateral ANOVA followed by ounnetl as a posteriori test. The ounnetl test was applied to compare against vehicle control. The differences were considered statistically significant at the p lt level; 0.05.

Automated evaluation of food intake in mice

NMRI female mice of a body weight between 20 and 30 g were used. The mice were adapted to the housing conditions for at least one week and for at least one day they were individually housed in the evaluation equipment, when the baseline data were recorded simultaneously . On the day of the study, the test product was administered subcutaneously close to the phase of switching off the lights (at 12 o'clock the lights were turned off and the evaluation of the feed intake started directly afterwards). The evaluation included continuous monitoring for 22 hours , while the data were processed as an average during every 30 min. It was possible to repeat this procedure for several days.The restriction of the evaluation to 22 hours was for practical reasons to allow re-weighing the animals, refilling of food and water and drug administration between procedures. The results could be evaluated as data accumulated during 22 hours or differentiated in intervals of 30 min. Comparable data can be obtained for both female and male mice.

Statistical analyzes were performed with Everstat 6.0 by bilateral ANOVA in repeated measures and later analysis of ounnetl. The differences were considered statistically significant at the level of p lt; 0.05.

Acute and subchronic effects of exendin-4 derivatives after subcutaneous treatment on blood glucose and body weight in female C57BU6NCrl mice with diet-induced obesity (DIO)

18 months with a ketogenic diet (method 1)

Female C57BU6NCrI mice were housed in groups in a specific pathogen-free barrier facility in a 12 h light / dark cycle with free access to water and a ketogenic diet. After 18 weeks with a ketogenic diet, the mice were stratified into treatment groups (n = 8), so that each group had a similar average body weight. A group of the same age with access at will to conventional feed was included as a conventional control group

Before the experiment, the mice were injected subcutaneously (sc) with vehicle solution and weighed for 3 days to acclimate them to the procedures.

1) Acute effect on blood glucose in DIO mice fed: initial blood samples were taken just before the first administration (sc) of vehicle (phosphate buffer solution) or exendin-4 derivatives at doses of 10, 30 And 100 1J9fkg (dissolved in phosphate buffer), respectively. The volume of administration was 5 ml / kg. The animals had access to water and their corresponding diet during the experiment, the food consumption was determined at all time points of the blood sampling. Blood glucose levels were measured at t = 0.5 h, t = 1 h, t = 2 h, t = 4 h, t = 6 h, t = 8 h and t = 24 h (method: d-glucose) hexokinase, hemolysed, AU640 Beckman Coulter). Blood sampling was performed by incision in the tail without anesthesia.

2) Subchronic effect on body weight: all animals were treated once a day sc in the evening, at the end of the light phase (at 12 o'clock the light is switched on) with both vehicle and exendin-4 derivatives to the doses mentioned above for 4 weeks. Body weight was recorded daily. On days 6 and 28, the total adipose mass was measured by nuclear magnetic resonance (NMR) using a Bruker minispec (Ettlingen, Germany).

14 weeks of previous feeding with a ketogenic diet (method 2)

Female C57BL / 6NCr1 mice were housed in groups in a specific pathogen-free barrier facility in a 12 h light / dark cycle with free access to water and a ketogenic diet. After 14 weeks with the ketogenic diet, the mice were stratified into treatment groups (n = 8), so that each group had a similar average body weight. A group of the same age was included with an access at will to standard feed and water as a standard control group.

Before the experiment, the mice were injected subcutaneously (sc) with vehicle solution and weighed for 3 days to acclimate them to the procedures. Subchronic effect on body weight: all animals were treated once a day sc at the end of the evening, at the end of the light phase (LO 12:12) with either vehicle or exendin-4 derivatives at the dose previously mentioned for 3 weeks. Body weight was recorded daily.

Statistical analyzes were performed with Everstat 6.0 by repeated measures of bilateral ANOVA and a posteriori analysis of Dunnelt (glucose profile) and unilateral ANOVA, followed by a posteriori Dunnelt test (body weight, body fat). The differences compared to DIO control mice treated with vehicle were considered statistically significant at the p lt level; 0.05

Acute and subchronic effects of exendin-4 derivatives after subcutaneous treatment on blood glucose and HbA1c in female diabetic dbfdb mice deficient in leptin receptors (method 3)

BKS.Cg-m + 1 + mice Leprdb / J (dbfdb) and BKS.Cg-m +1 + Leprdb / + (thin control) female from Charles River Laboratories, Germany, were obtained at an age of 9 -10 weeks. The animals were housed in groups in a specific pathogen-free barrier facility in a 12 h light / dark cycle with free access to water and conventional rodent feed. After 1 week of acclimatization, blood samples were taken from the tail without anesthesia and the blood glucose was determined (method: d-glucose hexokinase, hemolysed, AU640 Beckman Couller) and the level of HbA1c (method: hemolysed, Cobas6000 c501 , Rache Diagnostics, Germany)

HbA1c is a glycosylated form of hemoglobin whose level reflects the average level of glucose to which the erythrocyte has been exposed during its lifetime. In mice, HbA1c is a relevant biomarker for the average blood glucose level during the preceding 4 weeks (erythrocyte life in mice -47 days).

Db / db mice were stratified to treatment groups (n = 8), so that each group had similar blood glucose levels and similar HbA1c

1) Acute effect on blood glucose in fed db / db mice: initial blood samples were taken just before the first administration (sc) of vehicle (phosphate buffer solution) or exendin-4 derivatives at doses of 3 , 10 and 100 IJgfkg (dissolved in phosphate buffer), respectively. The volume of administration was 5 mllkg. The animals had access to water and feed during the experiment, food consumption was determined at all time points of the blood sampling. The blood glucose levels were measured at t = 0.5 h, t = 1 h, t = 2 h, t = 4 h, t = 6 h, t = 8 h and t = 24 h. Blood sampling was performed by incision in the tail without anesthesia. Comparable data can be obtained for both female and male mice.

2) Subchronic effect on blood glucose and HbA1c: all animals were treated once a day sc in the evening, at the end of the light phase (12 h the lights on), with either vehicle or derivatives of exendin-4 at the aforementioned doses for 4 weeks. At the end of the study, blood samples (tail, without anesthesia) were analyzed for glucose and HbA1c. Comparable data can be obtained for both female and male mice.

Statistical analyzes were performed with Everstat 6.0 by repeated measures of bilateral ANOVA and a posteriori analysis of Dunnetl. The differences compared to dbfdb control mice treated with vehicle were considered statistically significant at the p lt level; 0.05.

Effects of 4 weeks of treatment on glucose, HbA1c and oral glucose tolerance in female diabetic dbdb mice (method 4)

8-week-old female diabetic dbdb mice of average fasting glucose value of 14.5 mmolll and a body weight of 37-40 g were used. The mice were individually labeled and adapted to the housing conditions for at least one week.

7 days before the start of the study, the initial values ​​for non-fasting glucose and HbA1c were determined, 5 days before the start of the study, the mice were assigned to groups and cages (5 mice per cage, 10 per group) according to their values of HbA1c to ensure uniform distribution of lower and higher values ​​between groups (stratification)

The mice were treated for 4 weeks, by subcutaneous administration once a day, 3 hours before the dark phase (6 pm to 6 am). Blood samples were obtained from a tail tip incision for HbA1c on study day 21 and oral glucose tolerance was assessed in the 4th week. The oral glucose tolerance test was performed in the morning without prior administration of additional compound to evaluate mainly the effect of chronic treatment and the administration of the compound inferior to acute. The mice fasted for 4 hours before the administration of oral glucose (2 gl1lt; g, t = ° min). Blood samples were taken before administration of glucose and 15, 30, 60, 90, 120 and 180 min from here. The feed was returned after the last blood sample.

Statistical analyzes are performed with Everstat Version 6.0 based on SAS by unilateral ANOVA, followed by Dunnetl's subsequent test against vehicle control. The differences are considered statistically significant at the level of p lt; 0.05

Glucose reduction in female diabetic dbdb mice not fasting

Female diabetic dbdb mice with an average fasting glucose value of 20-22 mmolfl and a body weight of 42 g +/- 0.6 g (EEM) were used. The mice were individually labeled and adapted to the housing conditions for at least one week.

3-5 days before the start of the study, the mice were assigned to groups and cages (4 mice per cage, 8 per group, control group 16) according to their non-fasting glucose values ​​to ensure a homogeneous distribution of lower values and higher between groups (stratification). On the day of the study, the mice were weighed and dosed (t = O). Immediately prior to the administration of the compound, the feed was removed while the water was still available, and a first blood sample was drawn into an incision in the tail (initial level). Additional blood samples were removed from the tail incision at 30, 60, 90, 120, 240, 360, and 480 min.

The statistical analyzes are performed with Everstat Version 6.0 based on SAS by bilateral ANOVA in repeated measures, followed by the a posteriori test of Ounnel! against vehicle control. The differences are considered statistically significant at the level of p lt; 0.05

EXAMPLES

The invention is further illustrated by the following examples.

Example 1:

Synthesis of SEO ID NO: 8

Solid phase synthesis was carried out on Novabiochem Rink amine resin (4- (2 ', 4'-dimethoxyphenyl-Fmoc-aminomethyl) -phenoxyacetamido-norleucilaminomethyl) resin, 100-200 mesh, 0.34 loading mmolfg. The Fmoc synthesis strategy was applied with activation of HBTUfDIPEA. N-Boc-4- (Fmocamino) piperidine-4-carboxylic acid was used as the amino acid in the 20-position. Boc-Tyr (tBu) -OH and position 14 Fmoc-Lys (ivDde) ​​were used in the position. OH in the solid phase synthesis protocol The ivDde group was excised from the peptide on resin according to a modified procedure of the literature (SR Chhabra et al., Tetrahedron Lel. 39, (1998), 1603), using 4% hydrazine hydrated in OMF, then Palm-Glu (yOSu) -OtBu was coupled to the released amino group The peptide was cleaved from the resin with King's mixture (DS King, C.G. Fields, GB Fields, Inl. J. Peptide Protein Res. 36, 1990, 255-266). The crude product was purified by preparative HPLC on a Waters column (Sunfire, Prep C18) using a gradient of acetonitrile water (both buffers with 0.05% TFA). The purified peptide was analyzed by LC-MS (Method B). The deconvolution of the mass signals found under the peak with retention time 12.69 min revealed the mass of peptide 4618.71, which is in line with the expected value of 4619.21

Example 2 '

Synthesis of SEO ID NO: 11

Solid phase synthesis was carried out on Novabiochem Rink amine resin (4- (2 ', 4'-dimethoxyphenyl-Fmoc-aminomethyl) -phenoxyacetamido-norleucilaminomethyl) resin, 100-200 mesh, 0.34 loading mmollg. The 5 Fmoc synthesis strategy was applied with activation of HBTUfOIPEA. They were used in the position 1 Boc-Tyr (tBuquot; OH, in the position 14 Fmoc-Lys (ivDde) ​​-OH and in the position 20 Fmoc- (S) -MeLys (Boc) -OH in the synthesis protocol in solid phase The ivDde group was excised from the peptide on resin according to a modified procedure of the literature (SR Chhabra et al., Tetrahedron Lel., 39, (1998), 1603), using 4% hydrazine hydrated in OMF. Palm-Glu (yOSu) "OtBu was added to the released amino group, the peptide was excised from the resin with a mixture of

10 King (DS King, GC Fields, GB Fields, Inl. J. Peptide Protein Res. 36, 1990, 255-266). The crude product was purified by preparative HPLC on a Waters column (Sunfire, Prep C18) using a gradient of acetonitrile water (both buffers with 0.05% TFA). The purified peptide was analyzed by LC-MS (Method B). The deconvolution of the mass signals found under the peak with retention time of 12.88 min revealed the mass of peptide 4634.66, which is in line with the expected value of 4635.25

15 Example 3 '

Synthesis of SEO ID NO: 15

Solid phase synthesis was carried out on Novabiochem Rink amine resin (4- (2 ', 4'-dimethoxyphenyl-Fmoc-aminomethyl) -phenoxyacetamido-norleucilaminomethyl) resin, 100-200 mesh, 0.34 loading mmolfg. The Fmoc synthesis strategy was applied with activation of HBTUfOIPEA. They were used at position 1 Boc-Tyr (tBuquot; 20 OH Y at position 14 Fmoc-Lys (ivDde) ​​-OH and at position 20 Fmoc-alpha-methyl-ornithine {Boc) -OH in the synthesis protocol in solid phase. The ivOde group was cleaved from the peptide on resin according to a modified procedure of the literature (SR Chhabra et al., Tetrahedron Lel. 39, (1998), 1603), using 4% hydrazine hydrated in OMF. Stea-Glu (yOSu) -OlBu was then coupled to the liberated amino group. The peptide was cleaved from the resin with King's mixture (DS King, CG Fields, GB Fields, Inl. J. Peptide Protein Res. 36, 1990, 255-266). The raw product is

25 purified by preparative HPLC on a Waters column (Sunfire, Prep C18) using a gradient of acetonitrile water (both buffers with 0.1% TFA). The purified peptide was analyzed by LC-MS (Method B). The deconvolution of the mass signals found under the peak with retention time 12.90 min revealed the mass of peptide 4603.64, which is in line with the expected value of 4604.24

Analogously, the following peptides SEO ID NO: 8-17 were synthesized and characterized (Method AE), 30 see Table 5.

Table 5: List of peptides synthesized and comparison of calculated molecular weight against found.

<dl><dt>SEO ID NO ' </dt><dd>Masa cale Mass found </dd></dl>

<dl><dt>8 </dt><dd>4619.2 4618.7 </dd></dl>

<dl><dt>9 </dt><dd>4635.2 4634.8 </dd></dl>

<dl><dt>10 </dt><dd>4621, 2 4621, 1 </dd></dl>

<dl><dt>eleven </dt><dd>4635.2 4635.1 </dd></dl>

<dl><dt>12 </dt><dd>4649.3 4648.0 </dd></dl>

<dl><dt>13 </dt><dd>4634.3 4633.6 </dd></dl>

<dl><dt>,. </dt><dd>4649.3 4648.9 </dd></dl>

<dl><dt>fifteen </dt><dd>4604.2 4603.6 </dd></dl>

<dl><dt>16 </dt><dd>4548.2 4547.4 </dd></dl>

<dl><dt>1st </dt><dd>4252.7 4251.7 </dd></dl>

Unfixed comparison compound

Example 4: Chemical stability and solubility

The solubility and chemical stability of the peptide compounds were evaluated as described in Methods The results are given in Table 6.

Table 6: Chemical stability and solubility

<dl><dt>SEO ID NO: </dt><dd>Stability (pH 4.5) [%] Stability (pH 7.4) [%] Solubility (pH 4.5) [Jgfml] Solubility [Jgfml] (pH 7.4) </dd></dl>

<dl><dt>1 (Exendin-4) </dt><dd>100.0 77.5 933.6 1000 </dd></dl>

<dl><dt>8 </dt><dd>93.0 93.0 gt; 1000 gt; 1000 </dd></dl>

<dl><dt>eleven </dt><dd>100.0 99, 0 964.2 899.8 </dd></dl>

<dl><dt>12 </dt><dd>98.0 91, 0 gt; 1000 983.0 </dd></dl>

<dl><dt>fifteen </dt><dd>98.0 98.0 gt; 1000 gt; 1000 </dd></dl>

Example 5: In vitro data on the GLP-1, G1P and glucagon receptor

The potencies of peptide compounds in the GLP-1, GIP and glucagon receptors were determined

5 exposing cells expressing the human glucagon receptor (hGLUC R), human GIP (hGIP R) and the human GLP-1 receptor (hGLP-1 R) to the enumerated compounds at increasing concentrations and measuring cAMP formed as described in Methods

The results for the exendin-4 derivatives with activity in the human GIP receptor (hGIP R), human GLP-1 receptor (hGLP-1 R) and human glucagon receptor (hGLUC R) are shown in Table 7.

10 Table 7 CESO values ​​of exendin-4 peptide analogs in the GLP-1, GIP and glucagon receptors (indicated in pM)

<dl><dt>SEO ID NO · 8 9 10 11 12 13 14 15 16 </dt><dd>HESC of hGIP R [pM] 16.3 93.2 7.1 7.0.0.0.0.0.0.0.0.0.1.03.03.03.5 2.5 CESL of hGLP-1 R [pM] 4.7 13.5 4.9 5.1 11.6 13.6 7.2 16.1 4.1 HESEP of hGLUC R [pM] 34700.0 gt; 1000000 10400.0 3160.0 1.3 202.0 4730.0 4.0 19800.0 </dd></dl>

Comparison test

A selection of inventive exendin-4 derivatives comprising a functionalized amino acid has been tested

15 at position 14 against corresponding compounds having in this position 14 a 'non-functionalized' amino acid. The compounds by reference pairs and the corresponding CESO values ​​in the GLP-1 and GIP receptors (indicated in pM) are given in Table 8. As shown, the inventive exendin-4 derivatives show superior activity in comparison with compounds with a 'non-functionalized' amino acid at position 14

Table 8. Comparison of exendin-4 derivatives comprising a non-functionalized amino acid at position 14 against exendin-4 derivatives comprising an amino acid functionalized at position 14. CESO values ​​at GLP-1 and GIP receptors are indicated in pM. (K = lysine, L = leucine, and E-xS3 = {S) -4-carhoxy-4-hexadecanoylamino-butyryl-)

<dl><dt>SEO ID NO: </dt><dd>HOPE of hGIPR [pM] CESL of hGLP-1R [pM] rest at position 14 </dd></dl>

<dl><dt>eleven </dt><dd>7.0 5.1 K (yE xS3) </dd></dl>

<dl><dt>17 </dt><dd>103 5.9 L </dd></dl>

Example 6: Pharmacokinetic test Pharmacokinetic profiles were determined as described in Methods The calculated Tl12 and Cmax values ​​are shown in Table 9. Table 9. Pharmacokinetic profiles of exendin-4 derivatives.

<dl><dt>SEO ID NO · 11 15 </dt><dd>Mice (1 mgfkg) T12 [h] Cmax [ng / ml] 4.3 5940 2.9 3740 Minicerdos (0, 1 mg / kg) T12 [h] Cmax [ng / ml] 12.6 302 </dd></dl>

Example 7:

Subchronic effects of SE ID NO: 11 after subcutaneous treatment on body weight in mice

CS7BU6NCrl female with diet-induced obesity (DIO) (14 weeks of previous diet feeding

Ketogenic, method 2)

15 Female obese CS7BU6NCrl mice were treated for 3 weeks once a day subcutaneously at the end of the afternoon, before the end of the light phase (12 hours lights on) with 3 ~ g / kg AND 10 ~ g / kg of SEO ID NO: 11 or vehicle. Body weight was recorded daily

Treatment with SEQ ID NO: 11 reduced body weight, while the control group of the ketogenic diet even increased body weight (Fig. 1 and Table 10). The calculation of the relative body weight change from the

20 initial values ​​revealed a dose-dependent decrease in body weight, reaching 7.6% at 3 ~ gfkg and 17.4% at 10 ~ gfkg (Fig. 2), respectively.

Table 10. Weight change in DIO mice during a treatment period of 3 weeks (mean ± SEM) (represented as normalized at 0 mmol / L or min, Fig. 5), and the reduction of ABe under the glucose curve reached significance statistics at 3 and 10 IJg / kg compared to vehicle control (Fig. 6; plt; 0.05, unilateral ANOVA, followed by Dunnett's posterior test).

<dl><dt>Example (Dose) Diet control standard Ketogenic control diet SEO ID NO: 11 (3 ~ gfkg) SE ID NO: 11 (10 ~ g / kg) </dt><dd>Overall weight change (g) +0.3 ± 0.2 +2.7 tO, 3 -3.2 ± 0.6 -6.8 ± 0.7 </dd></dl>

<dl><dt>2S </dt><dd>Example 8: Effects of 4 weeks of treatment with SEO ID NO: 11 oral glucose in female diabetic dbdb mice (method 4) about glucose, HbA1c and tolerance to </dd></dl>

<dl><dt>Female dbdb mice received 3 and 10 ¡.gfkg of SEO ID NO: 11 or phosphate buffered saline (vehicle control) once a day, subcutaneously for four weeks. SEO ID NO: 11 REDUCED in a statistically significant way the non-fasting glucose compared to the vehicle control at doses of 3 and 10 μg / kg (Fig 3).</dt><dd /></dl>

<dl><dt>30 </dt><dd>In addition, SEO ID NO: 11 prevented an increase in HbA1c in a statistically significant manner compared to vehicle control at the dose of 3 and 10 ~ gfkg (Fig. 4; pL; O, OS, unilateral ANOVA, followed by Dunnett's post-test). Treatment with SEQ ID NO: 11 led to improved oral glucose tolerance</dd></dl>

Example 9: SEQ ID NO: 11, SEQ ID NO: 12 and SEQ ID NO: 15 on glucose reduction in dbdb mice 5 female diabetics not fasting

Female dbdb mice received 3 IJg / kg of SEQ ID NO: 11, SEQ ID NO: 12 and SEQ ID NO: 15 or phosphate buffered saline (vehicle control) subcutaneously, at the time O min_ The three compounds reduced immediately the glucose values ​​(initial level at 20-22 mmolll), reaching SEQ ID NO: 11 and SEQ ID NO: 12 the maximum effect of -11 mmolll and SEQ ID NO: 15 of -12 mmolll glucose reduction,

10 respectively, at 240 min and maintaining it until the end of the observation at 480 min (Fig. 7).

The three compounds achieved a significant statistical reduction of glucose compared to vehicle control from t: = 60 min until the end of the observation (plt; 0.05, bilateral ANOVA in repeated measurements, followed by a posteriori test of Ounnetl ).

Example 2: Effect of SEQ ID NO: 11 on gastric emptying and intestinal transit in female NMRI mice

15 Female NMRI mice, weighing on average 25-30 g, received 1, 10 AND 100 IJg / kg of SEQ ID NO: 11, or phosphate buffered saline (vehicle control) subcutaneously, 30 min before the administration of the colored bolus_ 30 min later, the evaluation of the stomach contents and intestinal transit was made (Fig 8)

Comparable data can be obtained for both female and male mice.

In these studies, SEQ ID NO: 11 reduced intestinal transit by 44, 68 and 69% (plt; 0.0001) and increased the content

20 gastric remaining 17, 97 and 106% (plt; 0.0001 versus vehicle control, unilateral ANOVA, followed by the posterior test of Ounnell), respectively.

Table 11 · Sequences

<dl><dt>SEQ ID NO: </dt><dd>sequence </dd></dl>

<dl><dt>1 </dt><dd>HGEGT -FT -SDLSKQMEEEAVRLFIEW -LK-NGGPSSGAPPPS-NH2 </dd></dl>

<dl><dt>2 </dt><dd>HAEGT -FTSDVSSy -L -EGQAAKEFIAW -LV-KGR-NH2 </dd></dl>

<dl><dt>3 </dt><dd>HAEGTF -T -SDVSSy -L -EGQAAK (VE-x53) -EF-IA-WLVRGRG </dd></dl>

<dl><dt>4 </dt><dd>and AE GTFI SDYSI AM DKI HQQ D FV NW L-LA-QKG -KKNDW -K -HNITQ </dd></dl>

<dl><dt>5 </dt><dd>HSQGT -FT -SDy -SKy -L -DSRRAQDFVQ-WL-MNT </dd></dl>

<dl><dt>6 </dt><dd>Y -GEGTFT -SDLSIQMEEEAVRLFIEW -L-KN-GGP - $ - $ - GAPPPS-NH2 </dd></dl>

<dl><dt>7 </dt><dd>yAEGT -FT - $ - DVSIy -L -EGQAAKEFIAW -L -VK · G · R · N H2 </dd></dl>

<dl><dt>8 </dt><dd>Y-AI bE -GT -F -T -SDL -SIQK (VE-x53) -EERAAP ip-EF-IEWLKNTGPSSGAPPPS-NH2 </dd></dl>

<dl><dt>SEQ ID NO · </dt><dd>sequence </dd></dl>

<dl><dt>9 </dt><dd>y -A lb-EGT -FT -SDl -SIQK (yE -x53) -EERAA (R) M ys-E -F -1-E -W -l -KNTGPSSGAPPP -SN H2 </dd></dl>

<dl><dt>10 </dt><dd>And -Aib-EGT -FTSDl -SIQK (yE-x53) -EE -RAA (S) MeOrn-E -F -1-E -W -lK -NTGP -SSGAP -PP -S-NH2 </dd></dl>

<dl><dt>eleven </dt><dd>Y-Ai bEG -T -F -T -SDl -SIQK (yE-x53) -EERAA (S) Melys-E -F -1-E -W -lKNT -GP -SSGAP -PP -SN H 2 </dd></dl>

<dl><dt>12 </dt><dd>YA ib-QGT -FTSDl -SKQK (yE-x70) -DEQRA (S) M elys-E -F -IE -W -l -K -SG -GP -SSGAP -PP -SN H 2 </dd></dl>

<dl><dt>13 </dt><dd>Y -Aib-QGTFTSDL -SIQK (yE-x70) -DEQRA (R) MeL ys-E -F -1-E -W -LKSG -GPSSGAP -PP -SN H 2 </dd></dl>

<dl><dt>1. </dt><dd>Y -Aib-EGT -FTSDlSIQK (yE-x70) -DERAA (S) MeL ys-EFIEW -LKNlGPSSGAPPPS-NH2 </dd></dl>

<dl><dt>fifteen </dt><dd>and -Aib-QGTF -T -SDL -SIQK (yE-x70) -DEQRA (S) MeOrn-E -F -IEW -L -K -AGGP -SSGAP -P -P -SN H 2 </dd></dl>

<dl><dt>16 </dt><dd>Y-Aib-EGTFT-5-0-L-5-IQK (yE-x53) -EE -RAA (S) MeLys-E -F -IEW -L -KAGGP -SSG -AP -P -PS-NH2 </dd></dl>

<dl><dt>17 </dt><dd>And -Aib-EGT -FT -SDL -SIQlEERAA- (S) Melys-E -F-IE-W -l -KNTGPSSGAPPPSN H2 </dd></dl>

LIST OF SEQUENCES

lt; 11 0gt; Sanafi

5 lt; 120gt; GLPlIGIP or trigonal dual agonists of GLP1fGIPfglucagon

lt; 130gt; DE20121179

lt; 150g; EP12306647.4 10 lt; 151gt;

lt; 160gt; 17

lt; 170gt; Palentln version 3.5

lt; 210gt; 1

lt; 211gt; 39

lt; 212gt; PRT

lt; 213gt; Heloderma suspectum

5 lt; 220gt;

lt; 222gt; (39) .. (39)

lt; 223gt; Extreme and amidated

10 lt; 400gt; 1

His Gly Glu Gly Thr Phe Thr Ser Asp Leu Ser Lys Gln Met Glu Glu 1 5 10 15

Glu Ala Val ArQ Leu Phe Ile Glu Trp Leu Lys Asn Gly Gly Pro Ser 20 25 30

Ser Gly Ala Pro Pro Pro Ser 35

lt; 210gt; 2 15 lt; 211gt; 30

lt; 212gt; PRT

lt; 213gt; Sapiens oven

lt; 220gt; 20 lt; 221gt; MOD_RES

lt; 222gt; (30) .. (30)

lt; 223gt; Extreme and amidated

lt; 400gt; 2

HiS Ala Glu Gly Thr Phe Thr Ser Asp Val Ser Ser Tyr Leu Glu Gly 1 5 10 15

Gln Ala Ala Lys Glu Phe Ile Wing Trp Leu val Lys Gly Arq 20 2S 30

lt; 210gt; 3 lt; 211gt; 31

lt; 212gt; PRT

<dl><dt>lt; 213gt; Artificial sequence</dt><dd /></dl>

<dl><dt>5 10 </dt><dd>lt; 220gt; lt; 223gt; Artificial polypeptide lt; 220gt; lt; 221gt; RES MOD 222gt; (20) .. (20) lt; 223gt; Lys derivatized to N6 with (S) -4-carboxy-4-hexadecanoylamino-bulyryl; 400gt; 3</dd></dl>

<dl><dt>His Al.a 1 </dt><dd>Gl.u Gl.y Thr Phe Thr 5 Be Asp Val. 10 Be Being Tyr Leu Gl.u Gl.y 15 </dd></dl>

<dl><dt>Gl.n </dt><dd>To Als Lys Glu Phe 20 Il.e Ala Trp Leu Val. 25 Arq G1y Arg Gl.y 30 </dd></dl>

<dl><dt>1S </dt><dd>lt; 210gt; 4 lt; 21 1gt; 42 lt; 212gt; PRT &lt;213gt; Homo sapiens</dd></dl>

<dl><dt>twenty </dt><dd>lt; 400gt; 4</dd></dl>

<dl><dt>Tyr Al a 1 </dt><dd>Glu Gl and Thr Phe 5 Ile Ser Asp Tyr Ser 10 Ile Ala Met Asp Lys l.5 </dd></dl>

<dl><dt>Ile Hios </dt><dd>Gln Gln 20 A $ p Phe Val A $ n Trp Leu 25 Leu To Gln Lys Gly Lys 30 </dd></dl>

<dl><dt>Lys </dt><dd>Asn Asp Trp Lys His Asn 35 Ile Thr Gln 40 </dd></dl>

<dl><dt>25 </dt><dd>lt; 210gt; 5 lt; 211 gt; 29 lt; 212gt; PRT &lt;213gt; Homo sapiens</dd></dl>

<dl><dt>lt; 400gt; 5</dt><dd /></dl>

His Ser Gln Gly Thr Phe Thr Ser Asp Tyr Ser Lys Tyr Leu Asp Ser 1 5 10 15

Arq Arq Ala Gln Asp Phe Val Gln Trp Leu Met Asn Thr 20 25

lt; 210gt; 6

lt; 211gt; 39 5 lt; 212gt; PRT

lt; 213gt; Artificial sequence

lt; 220gt;

lt; 223gt; Artificial polypeptide

lt; 220gt;

lt; 221gt; MOD RES

lt; 222gt; (39) (39)

lt; 223gt; Extreme and amidated

lt; 400gt; 6

Tyr Gly Glu Gly Thr Phe Thr Ser Asp Leu Ser Ile Gln Met Glu Glu 1 5 10 15

Glu Ala Val Arg Leu Phe Ile Glu Trp Leu Lys Asn Gly Gly Pro Ser 20 25 30

Ser Gly Ala Pro Pro Pro Ser 35

20 lt; 210gt; 7

lt; 211gt; 30

lt; 212gt; PRT

lt; 213gt; Artificial sequence

25 lt; 220gt;

lt; 223gt; Artificial polypeptide

lt; 220gt;

lt; 221gt; MOD_RES 30 lt; 222gt; (30) __ (30)

<dl><dt>lt; 223gt; Extreme and amidated</dt><dd /></dl>

<dl><dt>lt; 400gt; 7</dt><dd /></dl>

<dl><dt>Tyr Ala Gl u 1 </dt><dd>Gly Thr Pha 5 Thr Ser Asp Val 10 HE Ila Tyr Lau Glu Gly 15 </dd></dl>

<dl><dt>5 </dt><dd>Gln Ala Ala Lys 20 G ~ u Phe Ile Ala Trp Leu Val 25 Lys Gly Arg 30 </dd></dl>

<dl><dt>10 </dt><dd>lt; 210gt; 8 lt; 21 1gt; 39 lt; 212gt; PRT &lt;213gt; Artificial sequence</dd></dl>

<dl><dt>lt; 220gt; lt; 223gt; Artificial polypeptide</dt><dd /></dl>

<dl><dt>1S </dt><dd>lt; 220gt; lt; 221gt; RES MOD 222gt; (2) ... (2) lt; 223gt; 2-Methylalanine</dd></dl>

<dl><dt>twenty </dt><dd>lt; 220gt; </dd></dl>

<dl><dt>lt; 222gt; (14) .. (14) lt; 223gt; Lys derivatized in N6 with (S) -4-carboxy-4-hexadecanoylamino-butyryl</dt><dd /></dl>

<dl><dt>25 </dt><dd>lt; 220gt; </dd></dl>

<dl><dt>lt; 222gt; (20) .. (20) lt; 223gt; Xaa is 4-aminopiperidine-4-carboxylic acid</dt><dd /></dl>

<dl><dt>30 </dt><dd>lt; 220gt; lt; 221gt; MOD_RES: 222gt; (39) .. (39) lt; 223gt; Extreme and amidated</dd></dl>

lt; 400gt; 8

Tyr Ala Glu Gly Thr Phe Thr Ser Asp Leu Ser Ile Gln Lys Glu Glu 1 5 10 15

Arq Ala Ala Xa Glu Phe Ile Glu Trp Leu Lys Asn Thr Gly Pro Ser

20 25 30

Ser Gly Ala Pro Pro Pro Ser 35

lt; 210gt; 9

lt; 211gt; 39

lt; 212gt; PRT

lt; 213gt; Artificial sequence

lt; 220gt;

lt; 223gt; Artificial polypeptide

lt; 220gt;

lt; 222gt; (2) .. (2)

lt; 223gt; 2-Methylalanine

lt; 220gt;

lt; 221gt; MOD RES

lt; 222gt; (14) .. (14)

lt; 223gt; Lys derivatized in N6 with (S) -4-carboxy-4-hexadecanoylamino-butyryl

lt; 220gt;

lt; 222gt; (20) .. (20)

lt; 223gt; (R) -alpha-methyl-lysine

lt; 220gt;

lt; 221gt; MOD_RES

lt; 222gt; (39) .. (39)

lt; 223gt; Extremely amidated lt; 400gt; 9

Tyr Ala Glu Gly Thr Phe Thr Ser Asp Leu Ser Ile Gln Lys Glu Glu 1 5 10 15

Arq Ala Ala Ly. Gl u Phe Ile Glquot; Trp Leu Ly. Asn Thr Gly Pro Ser 20 25 30

Ser Gly Al. Pro Pro Pro Ser 35

lt; 210gt; 10

lt; 211gt; 39

lt; 212gt; PRT

lt; 213gt; Artificial sequence

lt; 220gt;

lt; 223gt; Artificial polypeptide

lt; 220gt;

lt; 221gt; MOO_RES

lt; 222gt; (2) .. (2)

lt; 223gt; 2-Methylalanine

lt; 220gt;

lt; 221gt; MOO RES

lt; 222gt; (14) .. (14)

lt; 223gt; Lys derivatized in N6 with (S) -4-carboxy-4-hexadecanoylamino-butyryl

lt; 220gt;

lt; 221gt; MOO_RES

lt; 222gt; (20) .. (20)

lt; 223gt; Xaa is (S) -alpha-methyl-omitine

lt; 220gt;

lt; 221gt; MOO RES

lt; 222gt; (39) _quot; (39)

lt; 223gt; Extreme and amidated

lt; 400gt; 10

Tyr Ala Glu Gly Thr Phe Thr Ser Asp Leu Ser Ile Gln Lys Glu Glu 1 5 10 15

Arq Wing Ala Xaa Glu Phe rle Glu Trp Leu Lys Asn Thr Gly Pro Ser

20 25 30

Ser Gly Ala Pro P ~ o Pro Ser 35

lt; 210gt; 11

lt; 211gt; 39 5 lt; 212gt; PRT

lt; 213gt; Artificial sequence

lt; 220gt;

lt; 223gt; Artificial polypeptide

lt; 220gt;

lt; 221gt; MOO RES

lt; 222gt; (2) __ (2)

lt; 223gt; 2-Methylalanine

lt; 220gt;

lt; 222gt; (14) .. (14)

<dl><dt>lt; 223gt; </dt><dd>Lys derivatized to N6 with (S) -4-carboxy-4-hexadecanoylamino-butyryl 20 </dd></dl>

lt; 220gt;

lt; 221gt; MOO_RES

lt; 222gt; (20) .. (20)

<dl><dt>lt; 223gt; </dt><dd>(S) -alpha-me-1-lysine 25 </dd></dl>

lt; 220gt;

lt; 221gt; MOO RES

lt; 222gt; (39) .. (39)

<dl><dt>lt; 223gt; </dt><dd>Extreme and amidated 30 </dd></dl>

lt; 400gt; eleven

Tyr Ala Glu Gly Thr Phe Thr Ser Asp Leu Ser Ile Gln Lys 1 5 10

Arq Ala Ala Lys Glu Phe rle Glu Trp Leu Lys Asn Thr Gly

20 25 30

Ser Gly Ala Pro Pro Pro Ser 35

lt; 210gt; 12

lt; 211gt; 39 5 lt; 212gt; PRT

lt; 213gt; Artificial sequence

lt; 220gt;

lt; 223gt; Artificial peptide

lt; 220gt;

lt; 221gt; MOO RES

lt; 222gt; (2) .. (2)

lt; 223gt; 2-Methylalanine

lt; 220gt;

lt; 222gt; (14) .. (14)

lt; 223gt; Lys derivatized in N6 with (S) -4-carboxy-4-octadecanoylamino-butyryl

lt; 220gt;

lt; 221gt; MOO_RES

lt; 222gt; (20) .. (20)

lt; 223gt; (S) -alpha-me1yl-lysine

lt; 220gt;

lt; 221gt; MOO RES

lt; 222gt; (39) .. (39)

lt; 223gt; Extreme and amidated

lt; 400gt; 12

Glu Glu 15

Pro Being

Tyr Ala Gln Gly Thr Phe Thr Ser Asp Leu Ser Lys Gln Lys Asp Glu 1 5 10 15

Gln Arg Ala Lys Glu Phe Ile Glu Trp Leu Lys Ser Gly Gly Pro Ser

20 25 30

Ser Gly Ala Pro Pro Pro Ser 35

lt; 210gt; 13

lt; 211gt; 39 5 lt; 212gt; PRT

lt; 213gt; Artificial sequence

lt; 220gt;

lt; 223gt; Artificial polypeptide

lt; 220gt;

lt; 221gt; MOO RES

lt; 222gt; (2) .. (2)

lt; 223gt; 2-Methylalanine

lt; 220gt;

lt; 222gt; (14) .. (14)

lt; 223gt; Lys derivatized in N6 with (S) -4-carboxy-4-octadecanoylamino-butyryl

lt; 220gt;

lt; 221gt; MOO_RES

lt; 222gt; (20) .. (20)

lt; 223gt; (R) -alpha-methyl-lysine

lt; 220gt;

lt; 221gt; MOO RES

lt; 222gt; (39) .. (39)

lt; 223gt; Extreme and amidated

lt; 400gt; 13

Tyr Ala Gln Gly Thr Phe Thr Ser A8p Leu Ser Ile Gln Lys Asp Glu 1 5 10 15

Gln Arg Ala Lys G ~ u Phe Ile Glu Trp Leu Lys Ser Gly Gly Pro Ser 20 2S 30

Ser Gly Ala Pro P ~ o Pro Ser 35

lt; 210gt; 14

lt; 211gt; 39 5 lt; 212gt; PRT

lt; 213gt; Artificial sequence

lt; 220gt;

lt; 223gt; Artificial peptide

lt; 220gt;

lt; 221gt; MOO RES

lt; 222gt; (2) .. (2)

lt; 223gt; 2-Methylalanine

lt; 220gt;

lt; 222gt; (14) .. (14)

lt; 223gt; Lys derivatized in N6 with (S) -4-carboxy-4-octadecanoylamino-butyryl

lt; 220gt;

lt; 221gt; MOO_RES

lt; 222gt; (20) .. (20)

lt; 223gt; (S) -alpha-me1yl-lysine

lt; 220gt;

lt; 221gt; MOO RES

lt; 222gt; (39) .. (39)

lt; 223gt; Extreme and amidated

lt; 400gt; 14

Tyr Ala Glu Gly Thr Phe Thr Ser Asp Leu Ser Ile Gln Lys Asp Glu 1 5 10 15

Arq Ala Ala Lys Glu Phe rle Gl u Trp Leu Lys Asn Thr Gly Pro Ser

20 25 30

Ser Gly Ala Pro Pro Pro Ser 35

lt; 210gt; fifteen

lt; 211gt; 39 5 lt; 212gt; PRT

lt; 213gt; Artificial sequence

lt; 220gt;

lt; 223gt; Artificial peptide

lt; 220gt;

lt; 221gt; MOD RES

lt; 222gt; (2) .. (2)

lt; 223gt; 2-Methylalanine

lt; 220gt;

lt; 222gt; (14) .. (14)

lt; 223gt; Lys derivatized in N6 with (S) -4-carboxy-4-octadecanoylamino-butyryl

lt; 220gt;

lt; 222gt; (20) .. (20)

lt; 223gt; Xaa is (S) -alpha-methyl-omitine

lt; 220gt;

lt; 221 gt; MOD RES

lt; 222gt; (39) .. (39)

lt; 223gt; Extreme and amidated

lt; 400gt; fifteen

Tyr Ala Gln Gly Thr Phe Thr Ser A8p Leu Ser Ile Gln Lys Asp Glu 1 5 10 15

Gln Arg Ala Xaa Glu Phe Ile Glu Trp Leu Lys Wing Gly Gly Pro Ser

20 25 30

Ser Gly Ala Pro Pro Pro Ser 35

lt; 210gt; 16

lt; 211gt; 39 5 lt; 212gt; PRT

lt; 213gt; Artificial sequence

lt; 220gt;

lt; 223gt; Artificial polypeptide

lt; 220gt;

lt; 221gt; MOO RES

lt; 222gt; (2) .. (2)

lt; 223gt; 2-Methylalanine

lt; 220gt;

lt; 222gt; (14) .. (14)

lt; 223gt; Lys derivatized in N6 with (S) -4-carboxy-4-hexadecanoylamino-butyryl

lt; 220gt;

lt; 221gt; MOO_RES

lt; 222gt; (20) .. (20)

lt; 223gt; (S) -alpha-me1yl-lysine

lt; 220gt;

lt; 221gt; MOO RES

lt; 222gt; (39) .. (39)

lt; 223gt; Extreme and amidated

lt; 400gt; 16

Tyr Ala Glu Gly Thr Phe Thr Ser Asp Leu Ser Ile Gln Lys Glu Glu 1 5 10 15

Arq Ala Ala Lys Glu Phe rle Glu Trp Leu Lys Ala Gly Gly Pro Ser

20 25 30

Ser Gly Ala Pro P ~ o Pro Ser 35

lt; 210gt; 17

lt; 211gt; 39

5 lt; 212gt; PRT

lt; 213gt; Artificial sequence

lt; 220gt;

lt; 223gt; Artificial peptide

lt; 220gt;

lt; 221gt; MOD RES

lt; 222gt; (2) .. (2)

lt; 223gt; 2-Methylalanine

lt; 220gt;

lt; 222gt; (20) .. (20)

lt; 223gt; (S) -alpha-methyl-l isine

lt; 220gt;

lt; 222gt; (39) .. (39)

lt; 223gt; Extreme and safe

lt; 400gt; 17

Tyr Ala Glu Gly Thr Phe Thr Ser Asp Leu Ser Ile Gln Leu Glu Glu 1 5 10 15

Arq Wing Wing Lys Glu Phe ne Glu Trp Leu Lys Asn Thr G1y Pro Ser

20 25 30

Ser Gly Ala Pro PrO PrO Ser