RU2422461C1 - Exenatid analogues - Google Patents

Abstract

FIELD: medicine, pharmaceutics. ^ SUBSTANCE: invention refers to new Exenatid analogues of general formula H-HisGlyGluGlyThrPheThrSerAspLeuSerLysGlnNleGluGluGluAlaValArgLeuPhe-IleGluTrpLeuLysAsnGlyGlyProSerSerGlyXProProProSer-ol where X is selected from L-Ala or D-Ala. The produced analogues show the stability higher than Exenatid, high insulinotropic activity, and also have high activity on renal sodition discharge without potassium ion loss, intensifis calcium and magnesium ion discharge, osmotically free water dishcarge. These properties can be effective for water-salt metabolism rebalancing along with glycemia normalisation. ^ EFFECT: production of the compositions which can be used for treatment of diabetes, and also for treatment and prevention of diabetic nephropathy and cardiac failure. ^ 3 cl, 7 dwg, 4 tbl, 5 ex

Description

The invention relates to medicine and can be used in pharmacology as medicines for the treatment of diabetes mellitus, as well as for the treatment and prevention of diabetic nephropathy and heart failure.

Diabetes mellitus is a group of metabolic diseases characterized by pathologically elevated levels of glucose in the blood, which occurs due to defects in insulin secretion, insulin action, or both defects. The number of patients with diabetes in the world at the end of 1999 was 120-140 million, and this number is projected to double by 2025 [1]. The diagnostic criterion for diabetes mellitus is an increase in glucose concentration in fasting venous and capillary blood plasma> 7.8 mmol / l or in whole venous or capillary blood> 6.7 mmol / l; 2 hours after glucose loading (75 g orally), the glucose level in the plasma of venous blood> 11.1 mmol / L (200 mg / 100 ml) and in the plasma of capillary blood> 12.2 mmol / L (220 mg / 100 ml) ; in whole venous blood> 10.0 (180 mg / 100 ml) and in whole capillary blood> 11.1 mmol / l (200 mg / 100 ml). The main goal of diabetes treatment is to maintain normal blood glucose levels. For this purpose, various drugs are known to be used, for example, sulfonylureas, metformin and insulin. Exenatide is one of the drugs, the effectiveness of the treatment of which non-insulin-dependent diabetes mellitus is proven in clinical trials in humans.

Exenatide is a polypeptide originally known as exendin-4 (exendin-4), a method for the isolation of which from the lizard venom Heloderma suspectum and the amino acid sequence HGEGTFTSDL SKQMEEEAVR LFIEWLKNGG PSSGAPPPS-NH ₂ were first published in April 1992 [2]. Exendin-4 is a member of the incretin peptide family and has a wide range of activities, such as increasing insulin secretion, enhancing glucose processing, suppressing appetite, slowing the movement of food from the stomach, etc.

Exenatide is known for its use: for example, US Pat. No. 5,424,286 [3] discloses the use of exenatide (also called exendin-4) to stimulate the release of insulin with an efficiency greater than the endogenous hormone GLP-1. In the description of the invention, the above examples confirm that exenatide is effective for the treatment of diabetes mellitus.

Later, other therapeutic effects of exenatide were found: weight loss [4], decreased motility and slower gastric emptying [5, 6], as well as the inotropic and diuretic effect of exenatide [7, 8, 9]. The use of exenatide for the treatment of polycystic ovary [10, 11, 12], the treatment and prevention of nephropathy [13, 14], the prevention and treatment of cardiac arrhythmia [15], for the differentiation of bone marrow cells [16], and the treatment of diabetes in pregnant women [17] , 18], for modulating the level of triglycerides and treating dyslipidemia [19, 20], suppressing the secretion of glucagon [21, 22], for differentiating non-insulin-producing cells into insulin-producing [23]. Application [24] describes the use of exenatide, agonists and antagonists of exenatide to affect the central nervous system. Patents [25, 26] describe the use of exenatide and its agonists to reduce food absorption. Application [27] describes stabilized exenatide compounds. Patents [28, 29] describe new uses of exenatide (exendin and exendin agonists).

However, of all the therapeutic effects of exenatide described above, the main in its use is the treatment of non-insulin-dependent diabetes mellitus. However, its use has a number of significant drawbacks, such as the need for frequent use in the form of injections (twice a day daily), the high cost of exenatide, and the presence of a number of side effects, for example, individual intolerance and nausea. In addition, the therapeutic effectiveness of exenatide is limited by its high toxicity, and the achievement of an effective reduction (normalization) of glucose in the blood, the first and main goal of treating diabetes, is limited by the inability to use high doses of exenatide.

Therefore, there is the task of creating new exenatide analogues that have lower cost, greater efficacy, and without the side effects of exenatide.

Applications [30, 31, 32] describe new pharmaceutical compositions using exenatide (exendin). The main disadvantage of the pharmaceutical compositions described in them is the introduction of additional substances in the dosage form, which leads to an increase in the cost of such drugs.

Applications [33, 34] describe modified exendin and exendin agonists. Applications [35, 36] describe novel exendin agonist compounds. Application [37] describes new peptide analogues of GLP-1 and exenatide. Applications [38, 39] describe novel derivatives of the insulinotropic exenatide peptide (exendin-4). Application [40] describes exendin and exendin agonists of prolonged action. Application [41] describes new peptide agonists with GLP-1 and exenatide activity. Application [42] describes a new analog of exenatide. Application [43] describes new exenatide analogues for the treatment of diabetes mellitus. Application [44] describes new derivatives of exenatide. Application [45] describes new exenatide analogues. Applications [46, 47] describe novel exenatide agonist compounds. Application [48] describes new varieties of exenatide for the treatment of diabetes. The main disadvantage of the exenatide analogues described in these patents is the possibility of enzymatic cleavage at the C-terminal amino acid sequence.

Application [49] describes new analogues of the GLP-1 peptide and exenatide. Applications [50, 51] describe a new analog of exenatide with effects on the brain. The main disadvantage of these analogues is the use of peptide drugs in higher doses than exenatide itself.

Application [52] describes new peptides, exenatide analogues for lowering blood glucose levels. The application [53] describes new insulinotropic peptides of prolonged action, analogues of exenatide. Application [54] describes new peptides for lowering blood glucose levels, exenatide analogues. The main disadvantage of the exenatide analogues described in these patents is the need for additional modifications of the peptide molecule, additional stages of the synthesis of such drugs.

The closest analogue of the claimed invention is the International application (WO - publication: WO 2009/011544 A; Korean patent) [55], selected as a prototype, in which the synthesis of exenatide analogues is common with the claimed invention.

The main disadvantage of the known invention is the need to use in the synthesis of non-natural synthetic acids, such as desamino-histidine, β-hydroxy-imidazo-propionic acid, β-carboxy-imidazo-propionic acid, imidazo-acetic acid and dimethyl-histidine, which complicates the synthesis scheme and leads to an increase in the cost of a drug based on such exenatide derivatives.

The claimed invention describes exenatide analogues with amino acid substitutions at positions 14 and 35, as well as those having a modification of the amino acid residue at position 39.

It is known that peptide molecules in blood plasma undergo enzymatic cleavage, which leads to a noticeable loss of biological activity. To prevent this process, in particular, in the peptide, some L-amino acids are replaced by D-amino acids ([56] - replacement in GLP-1 Ala (8) with D-Ala for resistance to DPP-IV). Therefore, substitutions of certain L-amino acids with D-amino acids in exenatide will lead to greater resistance to enzymatic cleavage.

An aqueous solution of exenatide has a slightly acid reaction (pH <5) (see Pharmacopoeia article on BAETA). In acidic aqueous solutions, peptide molecules are more susceptible to various chemical modifications, such as, for example, deamidation and hydrolysis. Therefore, a decrease in the number of amide groups in the molecule will lead to greater stability of the drug.

US patent application [57] describes exenatide degradation products having oxidized methionine in position 14 in their structure. An exenatide analogue is also provided with Met (14) replaced by Leu (14) [58], but this peptide is less active and has more short lifetime compared with exenatide. Therefore, the search for other analogues with the replacement of methionine at position 14 is relevant.

In this case, it is desirable that the new stable analogs of Exenatide have greater therapeutic capabilities in comparison with the already known analogues. In addition, such compounds will be preferred for practical use if they are stable during storage and use. In some drug delivery devices, such as pen syringes or hydrogel implants, the peptides should not be degraded when stored in aqueous solutions or in a humid environment for several months.

Changes in the primary structure of peptides, which lead to an increase in the stability of these drugs, can also lead to serious changes in their biological properties, for example, binding to specific receptors, activation of receptors, etc. Each of these properties can affect the change in activity of these compounds. New stable analogs of exenatide should be no less effective than exenatide.

New exenatide agonists were obtained using a single synthetic scheme suitable for the preparation of various analogues of this peptide.

The technical result of the claimed invention is the creation of new, more stable, exenatide analogues having no less biological activity than the prototype, which can become the basis for use as medicines. The obtained analogues have a high ability to normalize the concentration of glucose in the blood after glucose loading, and also have high activity in relation to the excretion of sodium ions by the kidney without loss of potassium ions, enhance the release of calcium and magnesium ions, the release of osmotically free water, which has not been described previously for other known exenatide analogues. These properties may be useful for restoring water-salt metabolism along with the normalization of glycemia.

The technical result in the claimed invention is achieved by using compounds of the formula:

H-HisGlyGluGlyThrPheThrSerAspLeuSerLysGlnNleGluGluGluAlaValArgLeuPheIleGlu-TrpLeuLysAsnGlyGIyProSerSerGlyXProProProSer-ol with insulinotropic and diuretic effects.

In addition, the technical result is achieved by the fact that in a compound having insulinotropic and diuretic action, L-Ala is used: H-HisGlyGluGlyThrPheThrSerAspLeuSerLysGlnNleGluGluGluAlaValArgLeuPheIleGlu-TrpLeuLysAsnGlGProGlyGlyGlyProly

In addition, the technical result is achieved by the fact that in a compound having insulinotropic and diuretic action, D-Ala is used: H-HisGlyGluGlyThrPheThrSerAspLeuSerLysGlnNleGluGluGluAlaValArgLeuPheIleGlu-TrpLeuLysAsnGerGlyProPro.

The complete sequence of the claimed analogues I and II was obtained by carbodiimide condensation in a solution of the corresponding protected fragments 1-34 and 35-39. Fragment 1-34 common to these analogues was obtained by condensation of fragments 1-4 and 5-34 by the solid-phase convergent method on a 2-chlorotrityl chloride resin using DIC / HOBt. A prototype of the obtained peptide was released in a mixture of TFA: TIS: EDT: H ₂ O. The purity by HPLC was 85% of the peak of the molecular ion in the mass spectrum of MALDI-TOF corresponded to the calculated one (Figure 1), the results of amino acid analysis are presented in table 1.

Table 1 The results of the amino acid analysis of the peptide Fragment 1-34 Analogue I Analog II A1a 1.01 2.08 2.01 Arg 0.91 0.99 0.98 Asp 1.97 1.91 2.03 Glu 6.20 6.25 6.17 Gly 5.12 5.01 5.22 His 0.89 0.94 0.90 Ile 1.13 1.10 1.09 Leu 3.25 3.23 3.31 Lys 2.02 2.08 2.17 Nle 1.00 1.00 1.00 Phe 1.97 1.95 2.01 Pro 0.95 4.02 3.91 Ser 3.78 3.76 3.82 Thr 1.80 1.77 1.76 Trp 0.94 0.99 0.95 Val 1.15 1.16 1.09

Fragments 35-39 (H-AlaProProProSer (tBu) ol and H-D-AlaProProProSer (tBu) ol) were synthesized by the classical method in solution. The BocProProoOH dipeptide was obtained by reacting the succinimide ester of N-tert-butyloxycarbonylproline and the proline sodium salt in tetrahydrofuran-water mixture. BocProProProOH was prepared by reacting a dipeptide succinimide ester and a proline sodium salt. The tert-butyloxycarbonyl group was removed by the action of a solution of trifluoroacetic acid in dichloromethane. The reaction of succinimide ester of N-benzyloxycarbonylalanine or N-benzyloxycarbonyl-D-alanine and tripeptide trifluoroacetate in dioxane in the presence of diisopropylethylamine gave the corresponding tetrapeptides, which were used in the carbodiimide condensation with O-tert-butyl-serinol. The benzyloxycarbonyl group from the resulting compounds was removed by catalytic hydrogenolysis in methanol.

Fmoc-Ser (tBu) -ol was prepared according to a typical procedure for the reduction of a carboxyl group with sodium borohydride [59].

The fragments of H-AlaProProProSer (tBu) ol and H-D-AlaProProProSer (tBu) ol were purified using reverse phase HPLC. The purity of the peptides was more than 95%, the peaks of molecular ions in the mass spectra of MALDI-TOF corresponded to the calculated ones, the values of the chemical shifts of protons and carbons by NMR spectra are presented in table 2.

table 2 The values of chemical shifts of protons and carbons from NMR spectra H-AlaProProProSer-ol FROM H H-D-AlaProProProSer-ol C H Ala D-ala -CH- 47.45 4.19 -Ch- 47.82 3.98 -CH ₃ 14.74 1.38 -CH ₃ 15.92 1.19 Pro Pro -CH- 60.23 4.25 -CH- 60.23 4.25 60.30 4,59 60.30 4,59 60.46 4.64 60.46 4.64 -N-CH ₂ - 47.45 -N-CH ₂ - 47.45 47.57 3.40- 47.57 3.40- 47.87 3.75 47.87 3.75 -N-CH ₂ -CH ₂ - 24.43 1.8-2.00 -N-CH ₂ -CH ₂ - 24.43 (3) 1.8-2.00 (3) -CH-CH ₂ - 29.40 -CH-CH ₂ 29.40 27.68 1.65- 27.68 1.65- 27.81 2,35 27.81 2,35 Ser-ol Ser-ol -CH- 52.64 3.81 -CH- 52.64 3.81 -O-CH ₂ - 58.53 3.38- -O-CH ₂ - 58.53 3.38- 58.68 3.57 58.68 3.57

After processing the obtained protected sequences 1-39 with a mixture of TFA: TIS: EDT: H ₂ O (complete removal of the protective groups), purification was performed using reverse phase HPLC. To confirm the structures of the obtained compounds, an amino acid analysis was performed (Table 1) and the molecular weight was determined using mass spectrometry (MALDI-TOF) (Figs. 2 and 3).

The technical result is confirmed by numerous experimental studies on the synthesis of the claimed exenatide analogues and the study of their biological activity.

Example 1

To obtain analogue (I), a convergent solid-phase synthesis method and classical synthesis in solution were used. Exenatide's 39 amino acid sequence was fragmented. Methionine at position 14 was replaced by norleucine, serine amide at position 39 was replaced by serinol. Some fragments were synthesized by the solid-phase method using the Fmoc- / tBu-strategy on a 2-chlorotrityl chloride resin, fragment 35-39 was obtained both by the solid-phase method and by the classical method in solution. The final condensation of fragments 1-34 and 35-39 was carried out in solution, controlling the degree of the reaction using HPLC.

After deblocking in a mixture of TFA: TIS: EDT: H20 and purification by reverse phase HPLC, the target peptide was obtained with a purity of 98%. The peak of the molecular ion in the MALDI-TOF mass spectrum corresponded to the calculated one.

Example 2

To obtain analogue (II), a convergent solid-phase synthesis method and classical synthesis in solution were used. Exenatide's 39 amino acid sequence was fragmented. Methionine at position 14 was replaced by norleucine, L-alanine at position 35 was replaced by D-alanine, the serine amide at position 39 was replaced by serinol. Some fragments were synthesized by the solid-phase method using the Fmoc- / tBu-strategy on a 2-chlorotrityl chloride resin, fragment 35-39 was obtained both by the solid-phase method and by the classical method in solution. The final condensation of fragments 1-34 and 35-39 was carried out in solution, controlling the degree of the reaction using HPLC.

After deblocking in a mixture of TFA: TIS: EDT: H ₂ O and purification by reverse-phase HPLC, the target peptide with a purity of 97% was obtained. The peak of the molecular ion in the MALDI-TOF mass spectrum corresponded to the calculated one.

Example 3

Aqueous buffer solutions of compounds I and II were incubated at 40 ° C and pH 7 for 3 months. Aliquots of these solutions were periodically selected and analyzed by HPLC. Over the entire period of the study, the degradation of peptides was <3% (<10% during the month in the patent application [60). Thus, it was shown that compounds I and II are more stable than exenatide and exenatide analogues, previously known.

Example 4

Verification of the insulinotropic effect of compounds I and II.

The insulinotropic effect of analogues I and II was determined by the change in blood glucose level. The experiments were performed on Wistar rats (Rattus norvegicus var. Albino), 4-6 months old, body weight 150-220 g. In the morning the day before the experiment, the rats were fed as usual, and for the subsequent time until the experiment they given food with free access to water. The experiments were carried out in accordance with international standards for working with experimental animals and approved by the Ethics Committee of the IEFB RAS. A study of the hypoglycemic effect of drugs was carried out on anesthetized rats in order to exclude the influence of stressful effects during the experiment on blood glucose levels. For anesthesia, rats were injected intraperitoneally with 6 ml / kg body weight of a mixture of 0.75% nembutal and 0.37% chloralose. 30 minutes after the administration of anesthesia, a drop of capillary blood was taken from the tail in the rats and the glucose level was determined using test strips (Accu-Chek Go glucometer, Roche, Switzerland).

Exenatide and its analogues (14-Nle-35-L-Ala-39-Serinol-exenatide (I) and H-Nle-35-D-Ala-39-Seriol-exenatide (II)) were dissolved in a 0.9% solution sodium chloride to a concentration of 1-5 μg / ml and were injected intramuscularly in a volume of 1 ml / kg of body weight. The control group of animals was injected with the same volume of 0.9% sodium chloride solution. 15 minutes after drug administration, rats were injected intraperitoneally with 3 ml / kg of a 50% glucose solution (1.5 g glucose / kg body weight). Animals of the control group were injected intraperitoneally with 3 ml / kg of 0.9% sodium chloride solution. 15, 30, and 60 minutes after glucose administration, capillary blood samples were taken from rats from the tail and glucose concentration was determined using a glucometer.

Statistical processing was performed using the program Statistica 6.0. All data are presented as M ± σ. The significance of differences between the groups was evaluated by the ANOVA test and the Newman-Cales test for multiple comparisons (at p <0.05).

results

Table 3 The effect of exenatide on the concentration of glucose in rat capillary blood under glucose loading Series of experiments n 0 min 15 minutes 30 minutes 60 min The control 5 6.1 ± 0.4 5.1 ± 0.3 4.6 ± 0.4 5.0 ± 0.2 Glucose 9 6.2 ± 0.6 11.4 ± 1.8 ^§ 10.3 ± 1.1 ^§ 5.4 ± 0.7 glucose + exenatide 1 mcg / kg 6 6.1 ± 0.2 10.3 ± 1.4 ^§ 8.3 ± 1.8 ^§ 4.4 ± 0.6 glucose + analog I 1 μg / kg 5 6.5 ± 0.5 11.0 ± 1.2 ^§ 11.3 ± 0.7 ^§ 5.7 ± 0.6 glucose + exenatide 2 mcg / kg 10 6.4 ± 0.7 8.4 ± 1.6 * ^§ 6.6 ± 1.6 * ^§ 4.7 ± 0.6 glucose + analog I 2 μg / kg 5 5.6 ± 0.4 10.1 ± 0.8 ^§ 7.9 ± 1.7 ^§ 4.6 ± 0.3 glucose + analog II 2 μg / kg 5 6.5 ± 0.6 8.9 ± 1.3 * ^§ 6.5 ± 0.7 * ^§ 4.9 ± 0.9 glucose + analog I 5 μg / kg 5 6.5 ± 0.7 6.9 ± 0.3 * ^§ 5.4 ± 0.9 * 4.3 ± 1.0 ^§ - significance of differences with the control group, * - significance of differences with the group with the introduction of glucose without drugs (p <0.05).

Thus, it was shown that for compounds I and II, with greater stability, the specific activity in normalizing the concentration of glucose in the blood against the background of sugar load remains at the level of exenatide.

Example 5

Testing the effect of compounds I and II on renal function.

The effect of analogues I and II on renal function was determined by measuring ion excretion, diuretic activity, and osmoregulatory renal function. The experiments were conducted on non-anesthetized Wistar rats (Rattus norvegicus var. Albino), 4-6 months old, body weight 150-220 g. Exenatide analogues (14-Nle-35-L-Ala-39-Serinol-exenatide (I) and 14-Nle-35-D-Ala-39-Seriol-exenatide (II)) was dissolved in 0.9% sodium chloride solution and was administered intramuscularly at a dose of 2 μg / kg body weight. The control group animals were injected with 0.9% sodium chloride solution. Animals were placed in cage-cases with a wire bottom, through the openings of which urine flowed down a funnel into a test tube. Urine excretion was recorded during spontaneous urination, urine volume, the concentration of sodium, potassium, magnesium, calcium and osmolality in it were measured. Blood was taken at the end of experiments from the carotid artery under ether anesthesia.

In blood and urine samples, osmolality was determined by cryoscopy using a 3300 microosmometer (Advanced Instruments, Inc., USA), the concentration of sodium and potassium ions using a Corning-410 flame photometer (United Kingdom) in an air-propane flame, and magnesium and calcium ions were measured using air-acetylene flame on a Hitachi-508 atomic absorption spectrophotometer (Japan).

Statistical processing was performed using the program Statistica 6.0. All data are presented as M ± o. The significance of differences between the groups was evaluated by the ANOVA test and the Newman-Cales test for multiple comparisons (at p <0.05).

results

After injection of exenatide analogues into rats, a marked increase in diuresis (Figure 4) of excretion of osmotically active substances (Table 4) and sodium (Figure 5) was observed. The excretion of magnesium and calcium cations increased slightly (Table 4), while potassium excretion did not significantly differ from the level in the control (Figure 6).

Table 4. The effect of exenatide analogues (2 μg / kg, i / m) on diuresis and excretion of cations and water by the kidney of rats. Excretion for 2 hours of experiment ^§ drug I (n = 6) drug II (n = 7) control (n = 10) Urine Volume, ml 2.0 ± 0.9 * 2.4 ± 0.6 * 0.4 ± 0.1 Osmotically active substances, microcosm 812 ± 223 * 865 ± 189 * 288 ± 46 Sodium ions, mmol 259 ± 82 * 283 ± 71 * 40 ± 12 Potassium ions, micromol 38 ± 18 37 ± 11 25 ± 6 Magnesium ions, mmol 5.4 ± 3.0 * 5.3 ± 2.2 * 2.2 ± 0.4 Calcium ions, mmol 1.3 ± 0.8 * 1.3 ± 0.8 * 0.3 ± 0.1 Osmotically free water, ml -0.7 ± 0.2 -0.5 ± 0.2 -0.6 ± 0.1 * - significance of differences with the control group (p <0.05). ^§ - indicators are calculated per 100 g of body weight.

The total reabsorption of osmotically free water for 2 hours of experiments did not change. When analyzing the dynamics of excretion / reabsorption of osmotically free water, it was found that in the first 30 minutes of drug action, a significant decrease in the reabsorption of osmotically free water in the kidney and the appearance of osmotically free water in the urine were noted. After 45 minutes of the action of the injected peptides, the excretion of osmotically free water changes to its intensive reabsorption (Fig. 7).

The obtained research results showed that compounds I and II have a high selective ion-regulating and diuretic activity, which were not described for other known exenatide analogues. Clinically valuable is the increased excretion of fluid and sodium by the kidney in combination with the preservation of potassium in the body, as this favors a decrease in blood pressure and does not cause adverse side effects associated with hypokalemia.

Thus, the claimed analogues of exenatide I and II:

1) more stable than exenatide and exenatide analogues (patent [60]);

2) have high insulinotropic activity;

3) have high activity in relation to the excretion of sodium ions by the kidney, the excretion of water in combination with the preservation of potassium ions, the excretion of calcium and magnesium ions, which has not been described for other known exenatide analogues.

Bibliography:

1. Diabetes mellitus, WHO information. Fact Sheet No. 138, Reviewed November (1999)

2. Eng, J. et al., Isolation and Characterization of Exendin-4, an Exendin-3 Analogue, from Heloderma suspectum Venom. J. Biol. Chem., Vol. 267, Issue 11, 7402-7405, Apr, 1992.

3. US 5424286 Exendin-3 and exendin-4 polypeptides, and pharmaceutical compositions containing the same

4. US 2005043238 Exendin formulations for weight reduction

5. US 6858576 Method for regulating gastrointestinal motility

6. US 9800449 Use of exendins and agonists therefore for the reduction of food intake

7. US 2005037958 Inotropic and diuretic effects of GLP-1 and GLP-1 agonists

8. US 6703359 Inotropic and diuretic effects of exendin

9. JP 2002509078 Inotropic and diuretic effects of exendin and GLP-1

10. US 2004266678 Methods and compositions for treating polycystic ovary syndrome

11. MX 2008015640 Compounds for treatment of metabolic disorders

12. US 2009291100 Therapeutic agent for polycystic ovary syndrome (PCOS)

13.JP 2006520747 Methods and compositions for treating polycystic ovary syndrome

14. US 2004209803 Compositions for the treatment and prevention of nephropathy

15. AU 2003297356 Prevention and treatment of cardiac arrhythmias

16. AU 2003262628 Bone marrow cell differentiation

17. US 2004092443 Long-acting exendins and exendin agonists

18. US 2004023871 Use of exendins and agonists therefore for the treatment of gestational diabetes mellitus

19. JP 2003519667 Use of exendins and agonists therefore for modulation of triglyceride levels and treatment of dyslipidemia

20. US 2003036504 Use of exendins and agonists therefore for modulation of triglyceride levels and treatment of dyslipidemia

21. US 9410225 Method for regulating gastrointestinal motility

22. RU 2247575 Method for inhibiting glucagon

23.US 2007041951 Differentiation of non-insulin producing cells into insulin producing cells by GLP-1 or exendin-4 and uses this

24. PT 1019077 Novel exendin agonist compounds

25. LU 91342 Use of exendins and agonists therefore for the reduction of food intake

26. US 2002137666 Use of exendins and agonists therefore for the reduction of food intake

27. US 2006194719 Stabilized exendin-4 compounds

28. US 2003087821 Exendins, exendin agonists, and methods for their use

29. EP 1419783 Use of a composition containing an exendin or a compound derived therefrom and a pharmaceutical carrier

30. CA 2487269 Novel exendin agonist formulations and methods of administration this

31. EP 1419783 Use of a composition containing an exendin or a compound derived therefore and a pharmaceutical carrier

32. US 2003087820 Novel exendin agonist formulations and methods of administration this

33. US 0011814 Modified exendins and exendin agonists

34. CN 1372570 Modified exendin and exendin agonists

35. US 0000902 Novel exendin agonist formulations and methods of administration therefore

36. CN 1284755 Cell lead, cell assemly using the same and combined cell

37. US 2004242853 Glp-1 exendin-4 peptide analogs and uses this

38. CN 0200316 Derivatives of exendin

39. US 2004142866 Derivatives of the insulinotropic peptide exendin-4 and methods of production thereof

40. US 2004092443 Long-acting exendins and exendin agonists

41. US 2004106547 Novel peptide agonists of GLP-1 activity

42. CN 1381271 Application of insulinotropic peptide Exendin 4 analog and its salt

43.US 6767887 Exendin analogues, processes for their preparation and medicaments containing them

44. EP 1056775 GLP-1 derivatives of GLP-1 and exendin with protracted profile of action

45. US 2009215694 Lyophilized formulations of exendins and exendin agonist analogs

46. US 9824273 Novel exendin agonist compounds

47. US 9824210 Novel exendin agonist compounds

48. DE 19637230 Truncated versions of exendin peptide (s) for treating diabetes

49. US 0224141 GLP-1 exendin-4 analogs and uses this

50. US 0126616 A novel peptide with effects on cerebral health

51. US2002115605 Novel peptide with effects on cerebral health

52. EP 1196444 Peptides that lower blood glucose levels

53. US 20080318861 Mucosal delivery of stabilized formulations of exendin

54. EP 1076066 Peptides for lowering blood glucose levels

55. KR 2008004170 Insulinotropic peptide derivative its own N-terminal ammo acid is modified (prototype)

56. CA 2003001470 Modified GLP-1 peptides with increased biological potency

57. US 20060194719 Stabilized exendin-4 compounds

58. Hargrove et. al., Biological activity of AC3174, a peptide analog of exendin-4. Regulatory Peptides 141 (2007) 113-119

59. Tetrahedron Letters, Vol 32, No. 7, pp. 923-926,1991

60. CA 2008000537 Exendin analogs

Claims (3)

1. The compound of the formula:
H-HisGlyGluGlyThrPheThrSerAspLeuSerLysGlnNleGluGluGluAlaValArg-LeuPheIleGluTrpLeuLysAsnGlyGlyProSerSerGlyXProProProSer-ol,
where X is selected from L-Ala or D-Ala,
possessing insulinotropic and diuretic action. 2. The compound of the formula according to claim 1, where X represents L-Ala:
H-HisGlyGluGlyThrPheThrSerAspLeuSerLysGlnNleGluGluGluAlaValArg-LeuPheIleGluTrpLeuLysAsnGlyGlyProSerSerGlyAlaProProProSer-ol. 3. The compound of the formula according to claim 1, where X represents D-Ala:
H-HisGlyGluGlyThrPheThrSerAspLeuSerLysGlnNleGluGluGluAlaValArg-LeuPheIleGluTrpLeuLysAsnGlyGlyProSerSerGlyD-AlaProProProSer-ol.

Images