MXPA01001905A - Method for administering insulinotropic peptides - Google Patents

Abstract

The claimed invention relates to a method of administering glucagon-like peptide-1 molecules by inhalation, a method for treating diabetes by administering glucagon-like peptide-1 molecules by inhalation, and a method for treating hyperglycemia by administering glucagon-like peptide-1 molecules by inhalation.

Description

USE OF INSULINOTROPIC PEPTIDES TO MANUFACTURE MEDICATIONS THAT ARE ADMINISTERED BY INHALATION Field of the Invention This invention relates to methods for treating humans suffering from diabetes and insulin resistance. In particular, the invention relates to the pulmonary delivery of peptide-1 similar to glucagon (GLP-1) and analogs thereof for systemic absorption through the lungs to eliminate the need to administer compounds ant i i cos -diabét by injection .

Background of the Invention Glucagon-like peptide-1 was first identified in 1987 as an incretin hormone, a peptide secreted by the intestine due to food ingestion. Glucagon-like peptide-1 is secreted by the L cells of the intestine after it is processed by prototyping the 160 amino acid precursor protein, preproglucagon. Cutting preproglucagon first produces glucagon-like peptide-1, a 37-amino acid peptide that is poorly active. A subsequent cut of the REF link. DO NOT. 126841 ^ S &UJF "peptide between residues 6 and 7 yields the peptide-1 biologically active glucagon-like referred to as GLP-l (7-37) It should be noted that this specification uses the nomenclature scheme that has developed around this. By convention in the art, the amino terminus of GLP-I (7-37) has been assigned the number 7 and the carboxy terminus is the number 37. Approximately 80% of GLP-I (7-37) is assigned. synthesized is amidated at the C after removal of the terminal glycine residue in the l cells and metabolic effects biological produce GLP-l (7-37) free acid and amide, GLP-1 (7-36 ) NH2, are indistinguishable As used here, these two naturally occurring forms will be referred to collectively as GLP-1.

GLP-1 is known to stimulate insulin secretion (action insul rópica inot) causing glucose uptake by cells which decreases the levels of serum glucose (see, eg, Mojsov, S., In t. J Pep ti de Pro t ei n Res ea rch, 4_0: 333-343 (1992)). Numerous GLP-1 analogues and derivatives that demonstrate inotropic insul action are known in the art. It has also been shown that the N-terminal histidine residue (His 7) is very important for activity ^ ^^^^^^^^ ^^^^^^^^^^^^^^^^^^^^^^^ insulinotropic GLP-1 (Suzuki, S., et al. Diabetes Res.; Clinical Practice 5 (Supp.1): S30 (1988).

Multiple authors have demonstrated the link between laboratory and mammalian experimentation, particularly human, inotropic responses to the exogenous administration of GLP-1. See, e.g., Nauck, M.A., et al. , Diabetologia, 36: 741-744 (1993); Gutniak, M. , et al., New England J. of Medicine, 326 (20): 1316-1322 (1992); Nauck, M.A., et al. , J. Clin. Invest. , £ 1: 301-307 (1993); and Thorens, B., et al. , Diabetes, 42 .: 1219-1225 (1993)].

The GLP-1-based peptides hold great promise as alternatives for insulin therapy for patients with diabetes who have failed in sulphuretics. GLP-1 has been studied intensively by academic researchers, and this research has established the following for patients with type II diabetes who have failed in sulphureas: 1) GLP-1 stimulates insulin secretion, but only during hyperglia periods. The safety of GLP-1 compared with insulin is improved for this property of GLP-1 and for the observation that the amount of insulin secreted is proportional to the magnitude of the hyperglycemia. In addition, GLP-1 therapy will result in the pancreatic release of insulin and the first step of insulin action to the liver. This results in lower circulating insulin levels in the periphery compared to subcutaneous insulin injections. 2) GLP-1 suppresses glucagon secretion, and this, in addition to the release of insulin via the portal vein helps to suppress excessive hepatic glucose output in diabetic patients. 3) GLP-1 decreases the gastric release that is desirable because it extends the absorption of nutrients over a long period of time, decreasing the glucose peak after tprandia 1. 4) Several reports have suggested that GLP-1 could improve insulin sensitivity in peripheral tissues such as muscle and fat.

) Finally, GLP-1 has been shown to be a potential appetite regulator.

The mealtime use of GLP-1-based peptides offers several advantages over insulin therapy. Insulin therapy requires monitoring blood glucose, which is expensive and painful. Glucose dependence of GLP-1 provides an improved therapeutic window compared to insulin, and should minimize the need to monitor blood glucose. Weight gain can also be a problem with therapy intensive insulin, particularly in obese type II diabetic patients.

The therapeutic potential for native GLP-1 is further increased if its use is considered in patients with type I diabetes. A number of studies have demonstrated the effectiveness of native GLP-1 in the treatment of insulin-dependent diabetes mellitus. Similar to patients with type II diabetes, GLP-1 is effective in rapidly reducing hipergl icemia by means of its attic glucagonost properties. Additional studies have indicated that GLP-1 also reduces postprandial glycemic excursions in type I patients, most likely through a delay in gastric release. These observations indicate that GLP-1 could be useful as a treatment in type I and type II patients.

To date, the administration of clinically proven peptide hormones and like GLP-1 has been carried out by subcutaneous injection and which is inconvenient and unattractive. Therefore, many investigators have studied alternative routes for the administration of peptide hormones such as the oral, rectal, transdermal and nasal routes. However, in this manner, routes of administration have not resulted in clinically proven peptide hormone therapy.

It has been known for a number of years, that some proteins can be absorbed from the lung. For example, insulin administered by aerosol inhalation to the lung first was reported by Gaensslen in 1925. Despite the fact that a number of human and animal studies have shown that some insulin formulations can be absorbed through the lungs, the release Pulmonary peptide hormones have not been vigorously continued due to the very low bioavailability. Higher proteins, such as cytokines and factors of ? kM, gg > These growths, which are generally greater than 150 amino acid residues, are often easily absorbed by the cells that cover the alveolar regions of the lung. The pulmonary absorption of lower proteins is, however, much less predictable; by means of insulin (51 residues), calcitonin (32 residues) and parathyroid hormone (34 residues) have been reported to be absorbed systemically through the pulmonary route. See US Patent No. 5,607,915, incorporated herein by reference. Despite the systemic absorption by the lung of some small protein hormones, the pharmacodynamics associated with the pulmonary release of the peptides is unpredictable.

Thus, there is a need to provide a reliable pulmonary method for the release of GLP-1 and related analogues, because it would offer patients an attractive, noninvasive alternative to insulin. This need is particularly true given that insulin has a very narrow therapeutic index, while GLP-1 treatment offers a way to normalize blood glucose only in response to hyperglycemic conditions without the threat of hypoglycemia.

Not all protein hormones can be efficiently absorbed through the lungs, and there are many factors that affect it. The absorption of proteins in the lung is largely dependent on the physical characteristics of the protein. Thus, even though the pulmonary release of some protein hormones has been observed, the physical properties and short length of GLP-1 and some related peptides, it is unclear whether such peptides could be effectively released through the pulmonary route.

Efficient pulmonary release is dependent on the ability to release the protein to the deep alveolar epithelium of the lung. The protein particles that lodge in the epithelium of the upper duct are not absorbed to a significant degree due to the functions of the overlying mucus to trap, and then clean the debris by mucociliary transport above the duct. This mechanism is also a major contributor to low bioavailability. The degree to which proteins are not absorbed and instead are eliminated by these í »k ^^^^^^ Ai» ^ ¿i ^? If the routes depend on their solubility, size and other widely uncharacterized mechanisms.

Even though a peptide hormone can be released reproductively into the deep alveolar epithelium of the lung, it is difficult to predict whether it will be rapidly absorbed and transported into the blood. The absorption values for some proteins released through the lungs have been calculated and are in the range of fifteen minutes for the parathyroid hormone (1-34) at 48 hours for the glycosylated al -ant i t r ips.

In addition, there is a variety of endogenous peptidases in the lung, which can degrade the peptides before absorption. In this way, the longer the time to take a peptide particle to dissolve and absorb, the greater the opportunity for enzymatic inactivation. In this way, due to the small size of GLP-1 and its inherent susceptibility to certain enzymes, it was more surprising to find that an aerosolized GLP-1 analog could be released reproducibly and effectively through the lungs.

Brief Description of the Invention The present invention relates to a method for The present invention relates to administering a glucagon-like peptide-1 molecule, comprising administering an effective amount of the peptide to a patient in need thereof by pulmonary release. The present invention also relates to a method of treating diabetes, comprising administering an effective dosage of a glucagon-like peptide-1 to a patient in need thereof by pulmonary release. Another aspect of the invention relates to a method for treating hyperglia, comprising administering an effective dosage of a glucagon-like peptide-1 to a patient in need thereof by pulmonary release. Preferably, the glucagon-like peptide-1 molecule is released by inhalation and into the patient's lower passage.

Glucagon-like peptide-1 can be released in a vehicle, as a solution or suspension or as a dry powder, using any variety of devices suitable for administration by inhalation. Preferably, glucagon-like peptide-1 is released in an effective particle size to achieve the lower lung passages. . ^ ..to íd.A? ... .. * ~. ^^ Ja ^^^ fe ^^^^^^^^,. ^, ^^, ^^, Detailed Description of the Invention The term "GLP-1" refers to human glucagon-like peptide-1 whose sequences and structures are known in the art. See US Patent No. 5,120,712, which is incorporated herein by reference. As previously discussed, there are two native forms of human GLP-1, GLP-1 (7-37) and GLP-1 (7 -36) NH 2, which will be distinguished only when necessary. The term "GLP-1 analogue" is defined as a molecule having one or more substitutions, deletions, inversions or additions of amino acids, compared to GLP-1. Many of the GLP-5 1 analogs are known in the art and include, for example, GLP-1 (7-34) and GLP-1 (7-35), GLP-I (7-36), Val8-GLP - 1 (7 - 37), Gln9-GLP-1 (7-37), D-Gln9-GLP-1 (7-37), Thr16-Lys 16-GLP-1 (7-37) and Lys18-GLP- 1 (7-37). The preferred GLP-1 analogs are GLP-I (7-34) and GLP-I (7-35), which are described in US Pat. No. 5,118,666, which is incorporated herein by reference.

The term "GLP-1 derivative" is defined as a molecule having the amino acid sequence of GLP-1 5 * or GLP-1 analog, but having additionally the chemical modification of one or more of its side groups of amino acids, α-carbon atoms, terminal amino groups or terminal carboxylic acid group. A chemical modification includes, but is not limited to, adding chemical radicals, creating new bonds and removing chemical radicals. Modifications in the amino acid side groups include, without limitation, acylation of the e-amino groups of lysine, N-alkylation of arginine, histidine or lysine, alkylation of glutamic or aspartic carboxylic acid groups and deamination of glutamine or asparagine. Modifications of the amino terminus include, without limitation, the modifications des-amino, N-lower alkyl, N-di-lower alkyl and N-acyl. Modifications of the carboxy terminal group include, without limitation, the modifications amide, lower alkyl amide, dialkyl amide and lower alkyl ester. The lower alkyl is alkyl Cj-C ,,, In addition, one or more side groups, or terminal groups, could be protected by protecting groups known to those skilled in the art of protein chemistry. The α-carbon of an amino acid could be mono or dimet i side.

The term "GLP-1 molecule" means GLP-1 & £ ^ Éá | ßÉj | s > Analogue of LPG-1 or LPG derivative 1.

Another preferred group of GLP-1 analogs is defined by the formula: Ri-X-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Val-Ser-Ser-Tyr-Leu-Y-Gly-Gln-Ala-Ala-Lys-Z-Phe-Ile-Ala-Trp- Leu-Val-Lys-Gly-Arg-R2 (SEQ ID NO: 1) and pharmaceutically acceptable salts thereof, wherein: Rx is selected from the group consisting of L-histidine, D-histidine, desamino-hi st idine, 2-amino-histidine, b-hydroxy-st stine, homohi st idina , alpha-f luoromet i 1 -hi st idina and 1 fa-me ti 1 -hi st idina; X is selected from the group consisting of Ala, Gly, Val, Thr, lie and al-Fa-met-Ala; And it is selected from the group consisting of Glu, Gln, Ala, Thr, Ser and Gly; Z is selected from the group consisting of Glu, Gln, Ala, Thr, Ser and Gly; and R2 is selected from the group consisting of NH2 and Gly-OH; with the proviso that when Rx is His, X is Ala, Y is Glu and Z is Glu, R2 must be NH2.

Still another preferred group of compounds t ^ áiS? ? Í Í Í k k k St St St St St St St St St St St St St St St St St? «-. - '"consistent with the present invention is described in WO 91/11457 (US Patent No. 5,545,618, which is incorporated herein by reference) and consists essentially of GLP-1 (7-34), GLP-1 (7-35), GLP-1 (7-36) 5 or GLP-I (7-37), or the amide forms thereof, and the pharmaceutically acceptable salts thereof, which have at least one modification selected from the group consisting of: (a) substitution of glycine, serine, cysteine, threonine, asparagine, glutamine, tyrosine, alanine, valine, isoleucine, leucine, methionine, phenylalanine, arginine or D-lysine by lysine at position 26 and / or position 34; or the substitution of glycine, serine, cysteine, threonine, asparagine, glutamine, tyrosine, alanine, valine, isoleucine, leucine, methionine, phenylalanine, lysine or D-arginine by arginine in position 36; (b) substitution of an oxidation-resistant amino acid by tryptophan at position 31; (c) substitution of at least one of: tyrosine for valine at position 16; lysine by serine in the position 18; aspartic acid by glutamic acid in the position 21; serine by. glycine in position 22; arginine for glutamine in position 23; arginine by alanine in position 24; and glutamine by lysine at position 26; and 5 (d) substitution of at least one of: glycine, serine or cysteine for alanine at position 8; Aspartic acid, glycine, serine, cysteine, threonine, asparagine, glutamine, tyrosine, alanine, valine, isoleucine, leucine, metiotine or phenylalanine for glutamic acid in position 9; serine, cysteine, threonine, asparagine, glutamine, tyrosine, alanine, valine, isoleucine, leucine, methionine or phenylalanine by glycine in position 10; and glutamic acid by aspartic acid in position 15; Y (e) substitution of glycine, serine, cysteine, threonine, asparagine, glutamine, tyrosine, alanine, valine, isoleucine, leucine, methylline or phenylalanine or the D or N form acylated or alkylated from histidine by histidine at position 7; wherein, in the substitutions is (a), (b), (d) and (e), the substituted amino acids may be optionally in the D form and the amino acids substituted in the 7-position may optionally be in the form N acylated or N rented Because the enzyme, idipept idyl-peptidase IV (DPP IV), could be responsible for the rapid inactivation x n vx vo observed of GLP-1 administered [see, e.g., Mentlein, R., et al. , Eu r. J. Bx or ch em. , 214: 829-835 (1993)], administration of GLP-1 analogs and derivatives that are protected from DPP IV activity is preferred, and administration of Gly8-GLP-1 (7 -36) is most preferred. NH2, Va 18-GLP-1 (7-37) OH, α-methyl-Ala8-GLP-I (7-36) NH2yGly8-Gln21-GLP-I (7-37) OH or the pharmaceutically acceptable salts of the smos.

The use in the present invention of a molecule claimed in U.S. Pat. No. 5,188,666, which is incorporated herein by reference. Such a molecule is selected from the group consisting of a peptide having the amino acid sequence: His-Ala-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Val-Ser-Ser-Tyr-Leu-Glu-Gly-Gln-Ala-Ala-Lys-Glu-Phe-Ile-Ala-Trp- Leu-Val-X (SEQ ID NO: 2) where X is selected from the group consisting of Lys and Lys-Gly; and a peptide derivative, wherein the peptide is selected from the group consisting of: a pharmaceutically acceptable acid addition salt of the peptide; a pharmaceutically acceptable carboxylate salt of the peptide; a pharmaceutically acceptable lower alkyl ester of the peptide and a pharmaceutically acceptable amide of the peptide selected from the group consisting of amide, lower alkyl amide and lower dialkyl amide. Another preferred group of molecules for use in the present invention consists of the compounds described in U.S. Pat. No. 5,512,549 which is incorporated herein by reference, which has the formula: R'-Ala-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Val-Ser-Ser-Tyr-Leu-Glu-Gly-Gln-Ala-Ala-Xaa-Glu-Phe-Ile-Ala Trp-Leu-Val-Lys-Gly-Arg-R3 R2 (SEQ ID NO: 3) and the pharmaceutically acceptable salts thereof, wherein R1 is selected from the group consisting of 4-imide zopropioni lo, 4-imide zoace 11 lo or 4-imidazo-a, a dimet i 1 -ace ti lo; R2 is selected from the group that consists of unbranched C6-C10 acyl, or is absent; R3 is selected from the group consisting of Gly-OH or NH2; and, Xaa is Lys or Arg, could be used in the present invention The most preferred compounds of SEQ ID NO: 3 for use in the present invention are those in which Xaa is Arg and R2 is unbranched C6-C10 acyl.

The highly preferred compounds of SEQ ID NO: 3 for use in the present invention are those at Xaa is Arg, R2 is unbranched C6-C10 acyl and R3 is Gly-OH.

The most highly preferred compounds of the SEC ID NO: 3 for use in the present invention are those in which Xaa is Arg, R2 is unbranched C6-C10 acyl, R3 is Gly-OH, and R1 is 4-imido zopropioni lo.

The most preferred compound of SEQ ID NO: 3 for use in the present invention is that in which Xaa is Arg, R 2 is unbranched C 8 acyl, R 3 is Gly-OH, and R 1 is 4-zopropionylimide.

The use of Val8-GLP-1 (7-37) OH is highly preferred The present invention also provides a pharmaceutically acceptable salt thereof, as claimed in US Patent No. 5,705,483, which is incorporated herein by reference in the present invention.

Methods for preparing GLP-1, GLP-1 analogs or GLP-1 derivatives useful in the present invention are well known in the art and are easily within the purview of the chemists or biochemists of proteins skilled in the art. The amino acid portion of the active compound used in the present invention, or a precursor thereof, can be made either by chemical synthesis in solid phase, purification of GLP-1 molecules from natural sources or recombinant DNA technology. Organic routine synthesis techniques allow the alkylation and acylation of the GLP-1 derivatives.

The term "GLP-1-related compound" refers to any compound that falls within the definition of GLP-1, GLP-1 analog or GLP-1 derivative.

The term condoms' s e r e f e e r üigfe ^ compound added to a pharmaceutical formulation to act as an ant i-microbial agent. A parenteral formulation must satisfy the guidelines for condom effectiveness to be a commercially viable multi-use product. Among condoms known in the art to be effective and acceptable in parenteral formulations are benzalkonium chloride, benzethonium, chlorhexidine, phenol, m-cresol, benzyl alcohol, methyl paraben, chlorobutanol, o-cresol, p-cresol, chlorocresol, phenylmercuric nitrate, thimerosal, benzoic acid and various mixtures thereof. I will see . g. , Wallhauser, K., De ve l op. Bi or l. S t a n da rd, 24: 9-28 (Basel, S. Krager, 1974). The term "buffer" or "pharmaceutically acceptable buffer" refers to a compound that is known to be safe for use in protein formulations and has the effect of Control the pH of the formulation at the desired pH for the formulation. Pharmaceutically acceptable buffers for controlling pH at a moderately acidic pH at a moderately basic pH include, for example, such compounds as phosphate, acetate, citrate, TRIS, arginine or histidine.

The term "isotonicity agent" refers to a compound that is physiologically tolerated and imparts an appropriate tonicity to a formulation to prevent the net flow of water through the cell membrane. Compounds, such as glycerin, are commonly used for such purposes at known concentrations.

A group consisting of a peptide has the amino acid sequence: His-Ala-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Val-Ser-Ser-Tyr-Leu-Glu-Gly-Gln-Ala-Ala-Lys-Glu-Phe-Ile-Ala-Trp- Leu-Val-X (SEQ ID NO: 2) wherein X is selected from the group consisting of Lys and Lys-Gly; and a peptide derivative, wherein the peptide is selected from the group consisting of: a pharmaceutically acceptable acid addition salt of the peptide; a pharmaceutically acceptable carboxylate salt of the peptide; a pharmaceutically acceptable lower alkyl ester of the peptide and a pharmaceutically acceptable amide of the peptide selected from the group consisting of amide, lower alkyl amide and lower dialkyl amide Another preferred group of molecules for use in the present invention consists of the compounds described in U.S. Pat. No. 5,512,549 which is incorporated herein by reference, which has the formula: R'-Ala-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Val-Ser-Ser-Tyr-Leu-Glu-Gly-Gln-Ala-Ala-Xaa-Glu-Phe-Ile-Ala-Trp - 10 Leu-Val-Lys-Gly-Arg-R3 (SEQ ID NO: 3) R2 and pharmaceutically acceptable salts thereof, wherein R1 is selected from the group consisting of 4- 15 imide zopropioni lo, 4- in the level of insulin circulation followed by a rapid drop in blood glucose levels. 20 Different inhalation devices typically provide similar acoractance when comparing similar particle sizes and similar levels of deposition in the lung.

According to the invention, GLP-1 and the analogues and ^ k ^ .. ^ ^^ ^, ^: ^^ s ^^^ ^^ ,. ^ &st * ^. «*. ^. - * moa® saife iais.S.i. ^^ GLP-1 derivatives can be released by any of a variety of inhalation devices known in the art for the administration of a therapeutic agent by inhalation. These devices include metered dose inhalers, nebulizers, dry powder inhalers, sprays, and the like. Preferably, GLP-1 and analogs and GLP-1 derivatives are released by an inhaler or a dry powder atomizer. There are several desirable characteristics of an inhalation device for the administration of GLP-1 and GLP-1 analogs and derivatives. For example, the release by the inhalation device is advantageously reliable, reproducible and accurate. He The inhalation device should release small particles, e.g. less than about 10 μm average mass aerodynamic diameter (MMAD), preferably about 1-5 μm MMAD, for good breathability. Some specific examples of The commercially available inhalation devices, or those of the last stage of development, suitable for the practice of this invention are turbohaler ™ (Astra), Rotahaler® (Glaxo), Diskus® (Glaxo), Spiros ™ inhaler (Dura), devices that developed by Inhale Therapeutics, AERx ™ (Aradigm), r? ^ ^ J ^ '^ c-Á "-'-" • ^ - ^^ Sa.jaittfc. ^^ gg ^ m the Ultravent® nebulizer (Mallinckrodt), the Acorn II® nebulizer (Marquest Medical Products), the Ventolin® metered dose inhaler (Glaxo), the Spinhaler® powder inhaler (Fisons) or similar.

As those skilled in the art will recognize, the formulation of GLP-1 or GLP-1 analogues or derivatives, the amount of the formulation released and the duration of administration of a single dosage, depend on the type of inhalation device employed. For some aerosol release systems, such as nebulizers, the frequency of administration and the duration of time during which the system is activated will depend primarily on the concentration of the GLP-1 molecule in the aerosol. For example, short periods of administration at high concentrations of GLP-1 and analogs and GLP-1 derivatives can be used in the nebulizer solution. Devices, such as metered dose inhalers, can produce high concentrations of aerosol, and can be operated for short periods to release the desired amount of GLP-1 and analogs and GLP-1 derivatives. Devices such as powder inhalers release the active agent until a given charge is expelled.

^ ^^? £? T ^ ¿i? AÁ ^ device agent. In this type of inhaler, the amount of GLP-1 and analogs and derivatives of GLP-1 in a given amount of the powder determines the dosage released in a single administration.

The particle size of GLP-1 and GLP-1 analogs and derivatives in the formulation released by the inhalation device is critical with respect to the ability of the protein to deposit in the lungs, and preferably in the lower ducts or alveoli. Preferably, GLP-1 and GLP-1 analogs and derivatives are formulated such that at least about 10% of the released peptide is deposited in the lung, preferably about 10% to about 20%, or more. It is known that the maximum efficiency of lung deposition for humans breathing through the mouth is obtained with particle sizes of about 2 μm to about 3 μm MMAD. When the particle sizes are above about 5 μm MMAD, the lung deposition decreases substantially. Particle sizes below approximately 1 μm MMAD cause the lung deposition to decrease, and it becomes difficult to release particles with sufficient mass that are ». * ..? AS & Therapeutically effective. In this manner, the GLP-1 particles and GLP-1 analogs and derivatives released by inhalation, have a particle size preferably of less than about 10 μm MMAD, more preferably in the range of about 1 μm to about 5 μm MMAD, and more preferably in the range of about 2 μm to about 3 μm MMAD. The formulation of GLP-1 and GLP-1 analogs and derivatives is selected to produce the desired particle size in the chosen inhalation device.

Advantageously, for administration as a dry powder, GLP-1 and GLP-1 analogs and derivatives are prepared in a particulate form resulting in an emitted particle size of less than about 10 μm MMAD, preferably about 1 to about 5 μm MMAD, and more preferably about 2 μm to about 3 μm MMAD. The preferred particle size is effective to be released into the alveolus of the patient's lung. Preferably, the dry powder is largely composed of produced particles, so that a majority of the particles have a size in the desired range. Advantageously, at least about 50% * > ~ > ^? ^ UI ^ St? TKEISUf ^ ll ^ j? ÜU ?? ^ tVI &eJlIl ItA m of the dry powder is made of particles having a diameter of less than about 10 μm MMAD. Such formulations can be achieved by spray drying, milling or critical point condensation of a solution containing the particular GLP-1 molecule or other desired ingredients. Other methods also suitable for generating particles useful in the current invention are known in the art.

The particles are usually separated from a dry powder formulation in a container and then transported in the lung of a patient via a conveyor air stream. Typically, in current dry powder inhalers, the force to disperse the solid is provided exclusively by the inhalation of the patient. An appropriate dry powder inhaler is the Turbohaler® manufactured by Astra (Sodertalje, Sweden). In another type of inhaler, the air flow generated by the inhalation of the patient activates a driving motor that deagglomerates the particles of the GLP-1 molecule. The Dura Spiros® inhaler is such a device.

The formulations of GLP-1 and analogs and GLP-1 derivatives for the administration of an inhaler of dry powder typically includes a finely divided dry powder containing the peptide, but the powder may also include a bulk agent, carrier, excipient, other additive or the like. The additives can be included in a dry powder formulation of GLP-1 and analogs and GLP-1 derivatives, for example, to dilute the powder as required for the release of the particular powder inhaler, to facilitate the processing of the formulation, to provide the powder properties advantageous for the formulation, to facilitate the dispersion of the powder from the inhalation device, to stabilize the formulation (eg, antioxidants or buffering agents), to provide flavor to the formulation, or the like. Advantageously, the additive does not adversely affect the patient's passages. The GLP-1 and GLP-1 analogs and derivatives can be mixed with an additive at a molecular level or the solid formulation can include particles of the peptides mixed with or coated on the additive particles. Typical additives include mono, di and polysaccharides; alcohols of sugars and other polyols, such as, for example, lactose, glucose, raffinose, melezitose, lactitol, maltitol, trehalose, sucrose, mannitol, starch or combinations thereof; surfactants, ,. ^? ^^. . ^, ... _ ^^^^. ^ ...... - iiiÉll ^ such as sorbitoles, diphosphatidyl choline or lecithin; or similar. Typically, an additive, such as a bulking agent, is present in an effective amount for a purpose described above, frequently to about 50% to about 90% by weight of the formulation. Additional agents known in the art for protein formulation can also be included in the formulation.

In another aspect of the invention, an atomization including GLP-1 and analogs and GLP-1 derivatives can be produced by forcing a suspension or solution of the peptide through a nozzle under pressure. The size and configuration of the nozzle, the pressure applied and the liquid feed speed can be chosen to achieve the desired particle size and output. An electro-atomization can be produced, for example, by an electric field in connection with a capillary or nozzle feed.

Advantageously, the drops of GLP-1 and analogs and GLP-1 derivatives released by an atomizer have an inhaled droplet size of less than about 10 μm MMAD, preferably in the range of about 1 μm to about 5 μm MMAD, and more preferably about 2 μm a approximately 3 μm MMAD.

The formulations of GLP-1 and GLP-1 analogs and derivatives suitable for use with an atomizer 5 are typically from about 1 mg to about 20 mg of the peptide per ml of the solution. The formulation may include agents such as an excipient, a buffer, an isotonicity agent, a preservative, a surfactant and metal cations. The formulation may also include an excipient or agent to stabilize the peptide, such as a buffer, a reducing agent, a bulk protein or a carbohydrate. The volume proteins useful in the formulation of GLP-1 And GLP-1 analogues and derivatives include albumin, protamine or the like. Typical carbohydrates useful in the formulation of GLP-1 and analogs and GLP-1 derivatives include sucrose, mannitol, lactose, trehalose, glucose or the like. The GLP-1 formulations and GLP-1 analogs and derivatives may also include a surfactant, which may reduce or prevent surface induced aggregation of the peptide caused by atomization of the solution in the formation of an aerosol. Several can be used conventional surfactants, such as esters and polyoxyethylene fatty acid alcohols, and polyoxyethylene sorbitol fatty acid esters. The quantities, in general, will be in the range between 0.001 and 4% by weight of the formulation. Other surfactants may also be used, such as diphosphatidyl choline or lecithin. Especially preferred surfactants for the purposes of this invention are polyoxyethylene sorbitan monooleate, polysorbate 80, polysorbate 20 or the like. Additional agents known in the art for protein formulation can also be included in the formulation.

GLP-1 and GLP-1 analogues and derivatives can be administered by a nebulizer, such as a jet nebulizer or an ultrasonic nebulizer. Typically, in a jet nebulizer, a source of compressed air is used to create a high velocity air jet through a hole. As the gas expands beyond the nozzle, a low pressure region is created, which triggers a solution of the peptide through a capillary tube connected to a liquid reservoir. The liquid stream from the capillary tube is cut into unstable filaments and the droplets as they exit the tube, creating the aerosol. A range of configurations, flow rates and types of BHi.jBt-aaK-iaafc divisions can be used to achieve the desired development characteristics of a given jet nebulizer. In an ultrasonic nebulizer, high frequency electrical energy is used to create the vibrational, mechanical energy, which typically employs a piezoelectric transducer. This energy is transmitted to the peptide formulation either directly through a coupling fluid, creating an aerosol. Advantageously, the drops of GLP-1 and analogs and GLP-1 derivatives released by a nebulizer have a particle size of less than about 10 μm MMAD, preferably in the range of about 1 μm to about 5 μm MMAD, and more preferably about 2 μm to about 3 μm MMAD.

Formulations of GLP-1 and GLP-1 analogs and derivatives suitable for use with a nebulizer, either jet or ultrasonic, typically include an aqueous solution of the peptide at a concentration of about 1 mg to about 20 mg per ml of solution. The formulation may include agents, such as an excipient, a buffer, an ionicity agent, a preservative, a surfactant and a * s? t tS% tr "- divalent metal cation. The formulation may also include an excipient or agent to stabilize the peptide, such as a buffer, a reducing agent, a bulk protein or a carbohydrate. Volume proteins useful in the formulation of GLP-1 and analogs and GLP-1 derivatives include albumin, protamine or the like. Typical carbohydrates useful in the formulation of GLP-1 related proteins include sucrose, mannitol, lactose, trehalose, glucose or the like. The formulations of GLP-1 and GLP-1 analogs and derivatives can also include a surfactant, which can reduce or prevent surface induced aggregation of the peptide caused by atomization of the solution in the formation of an aerosol. Several conventional surfactants can be used, such as esters and alcohols of polyoxyethylene fatty acids, and esters of polyoxyethylene sorbital fatty acids. The quantities, in general, will be in the range between 0.001 and 4% by weight of the formulation. Other surfactants such as phosphatidyl choline or lecithin may also be used. Especially preferred surfactants for the purposes of this invention are polyoxyethylene sorbitan monooleate, polya sorbate 80, polysorbate 20 or the like. The additional agents known in The art for the formulation of a protein, such as molecules related to GLP-1, can also be included in the formulation.

Another aspect of the invention involves a metered dose inhaler (MDI). In this embodiment, a propellant, GLP-1 and analogs and derivatives of GLP-1, and any excipients or other additives are contained in a container as a mixture that includes a liquefied compressed gas. The actuation of the dosing valve releases the mixture as an aerosol, preferably containing the inhaled particles in the size range of less than 10 μm MMAD, preferably about 1 μm a About 5 μm MMAD, and more preferably about 2 μm to about 3 μm MMAD. The desired aerosol particle size can be obtained by using a formulation of GLP-1 and analogs and GLP-1 derivatives produced by various methods known to those skilled in the art, including jet grinding, spray drying, critical point condensation or the like. Preferred metered dose inhalers include those manufactured by 3M or Glaxo and which employ a hydrofluorocarbon propellant.

The formulations of GLP-1 and analogs and GLP-1 derivatives for use with a metered dose inhaler device, in general, will include a finely divided powder containing the peptide as a suspension in a non-aqueous medium, for example, suspended in a propeller with the help of a surfactant. The propellant could be any conventional material used for this purpose, such as chlorofluorocarbon, a hydrochlorofluorocarbon, a hydrofluorocarbon or a hydrocarbon, including trichlorofluoromethane, dichlorodi fluoromethane, di chlorotetra fluoroethanol and 1, 1, 1, 2 -tet f luoroethane, HFA-134 a (hydrofluoroalkane-134 a), HFA-227 (hydrofluoroal cano-227), or the like. Preferably, the propellant is a hydrofluorocarbon. The surfactant may be chosen to stabilize the GLP-1 molecule as a suspension in the propellant, to protect the active agent against mechanical degradation, and the like. Suitable surfactants include sorbitan trioleate, soy lecithin, oleic acid or the like. In some cases the aerosols in solution are preferred using solvents, such as ethanol. Other surfactants such as diphosphatidyl choline or lecithin may also be used. The additional agents known in the art for the ^^. ^, ^ k. .í ... ^ á ^ ^:, ^^^. ^ Í. ^ .. ^. ^^ A ^ .. ^ protein formulation can also be included in the formulation.

The present invention also relates to a pharmaceutical composition or formulation that includes GLP-1 and GLP-1 analogs and derivatives and suitable for administration by inhalation. According to the invention, GLP-1 and GLP-1 analogs and derivatives can be used for the manufacture of an appropriate formulation or medicament for administration by inhalation. The invention also relates to methods for the manufacture of formulations that include molecules related to GLP-1 in a form that is suitable for administration by inhalation. For example, a dry powder formulation can be manufactured in various ways, using conventional techniques. Particles in the appropriate size range for maximum deposition in the lower respiratory tract can be made by micronization, grinding, spray drying or the like. And a liquid formulation can be manufactured by dissolving the peptide in an appropriate solvent, such as water, at an appropriate pH, including buffers or other excipients.

The present invention could be better understood with reference to the following examples. These examples are intended to be representative of the specific embodiments of the invention, and are not intended to limit the scope of the invention.

EXAMPLES Val8-GLP-1 Serum Pharmacokinetics in Beaqle Dogs after Pulmonary Administration The GLP-1 analog, Va 18-GLP-1 (7-37) OH (SEQ ID NO: 4) was prepared in E co l using conventional recombinant DNA techniques and purified in homogeneity.

NH2-His-Val-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Val-Ser-Ser-Tyr-Leu-Glu-Gly-Gln-Ala-Ala-Lys-Glu-Phe-Ile-Ala Trp-Leu-Val-Lys-Gly-Arg-Gly-OH (SEQ ID NO: 4) A group of 6 female beagle dogs was exposed to inhaled Val8-GLP-1 for 15 minutes at an average aerosol concentration of 77.2 μg / L generated from a solution of Val8-GLP-1 in sterilized water. These animals were dosed with 100 μg / kg by subcutaneous administration approximately 1 week after inhalation exposures. The volume of lung ventilation, respiration rate and volume / minute were monitored before and throughout the exposure period. Blood was collected during the analysis of the plasma levels of Val8-GLP-1 at various time points after inhalation and subcutaneous administration. The bronchoalveolar lavage fluid (BAL) was collected approximately 4 hours post-exposure and analyzed for LDH, total protein, cell counts and leukocyte differentials.

The release of Val8-GLP-1 was tolerated well with an inhaled dosage of 1198 μg / kg and an estimated deposited lung dosage of 240 μg / kg. Subcutaneous administration of 100 μg / kg was also well tolerated by all animals. Inhalation and subcutaneous administration of Val8-GLP-1 was released using the formulated material.

There were no clinical observations related to the treatment; body weights were not adversely affected by Val8-GLP-1. I only know observed minor pulmonary effects. The increases in Ventilation volume and volume / minute were observed during exposures to inhalation for 15 minutes, but the results were highly variable. No significant changes were observed for LDH, red blood cell counts, leukocyte counts, neutrophils, lymphocytes, eosinophils, epithelial cells, macrophages, basophils or monocytes. There was a moderate increase in the toral protein after the aerosol release of Val8-GLP-1.

The results of this study show that there was good bioavailability of Val8-GLP-1 (40%, based on AUC) released to the lungs of beagle dogs by inhalation in relation to subcutaneous administration. Val8-GLP-1 was well tolerated for up to 15 minutes with minimal effects on the lungs at an average inhaled dose of 1198 μg / kg, which was a level of unobserved adverse effects (NOAEL) in this study.

Preparation of Dosing Solutions The solutions of Val8-GLP-1 were prepared on the days of dosing at concentrations of 0.5 mg / ml or 8 mg / ml in sterile water for subcutaneous administration and pulmonary administration, respectively. An additional solution of Val8-GLP-1 (8 mg / ml) was prepared at the end of the live phase, to determine the particle size distribution of aerosolized Val8-GLP-1. The solutions were filtered through a 0.22 micron container filter of lower protein binding. The pH of the solution was adjusted with sodium hydroxide solution to 7.47.

Test animals Six female Beagle dogs (Marshall Farms, North Rose, NY) were used in this study. Each animal was identified only by a five-digit animal number and a seven-digit tattoo number (located on the inside of the ear) recorded on its cage card. All animals were acclimated to their restraining sling before beginning the study. The weight range of the animals at the beginning of the study was 8.4 to 11.1 kg. The age of the animals at the beginning of the study was 33 to 37 weeks.

Housing and care of the test animal The animals were housed in pairs in stainless steel cages except on the days of exposure. Each animal was individually housed on the day of exposure to monitor its feeding regime. The rooms were adjusted automatically to maintain a temperature of 70 ° C and maintain the current temperature within ± 8'F of the set point. The environmental control system is designed to maintain a relative humidity of 20% and a maximum of 80%. The light was on for a 12-hour cycle, with lights on between 0600 and 1800 hours. Subsequently, the lights went out between 1800 and 0600 hours, except when the blood samples were collected. The animals were fed once daily with Hill's Science Diet. The animals were fasted for approximately 12 hours before exposure. Water was provided from the tap to d l i bi t um except during exposures.

Treatment groups and study duration All 6 dogs were exposed for 15 minutes to aerosolized Val8-GLP-1. The dosage of the white deposited lung for the exposures of Val8-GLP-1 was 200 μg / kg of body weight. Approximately 7 days after the pulmonary administration of Val8-GLP-1, all dogs were dosed with 100 μg / kg of Val8-GLP-1 body weight by subcutaneous administration.

Aerosol generation The aerosols were generated using a Respirgard II nebulizer with an entrance of approximately 6.5 5 L / min. The output of the generator flowed directly into the head dome.

Sampling of atmospheric concentration All sampling for total gravimetric concentrations was performed with type A / E glass fiber filters containing in-line filter clips (Gelman Instruments Co., Ann Arbor, MI). Sampling of the camera filters took performed at a nominal sampling rate of 1 L / min, calibrated with a portable mass air flow calibrator (model 830, Sierra Instruments, Carmel Valley, CA). The duration of the sampling was 15 minutes. The particle size analysis is performed with a Sierra Model 218K Ambient Cascade Impactor (Anderson Samplers, Inc., Atlanta, GA). The cascade sampling of the chamber was carried out at a nominal sampling rate of 3 L / min, calibrated with a portable mass air flow calibrator (model 830, Sierra Instruments, Carmel Valley, CA). sfc? a The duration of the sampling was 31 minutes. The filters were allowed to dry for approximately 30 minutes before being reweighed.

Determination ote the dosage The dosage of inhaled Val8-GLP-1 during a 15-minute exposure was estimated as follows: the mean volume / minute (mL) during the 15-minute exposure was multiplied by the duration of the exposure to produce the total air breathed (L ) during exposure to inhalation. This value was multiplied by the concentration of the aerosol (μg / L) to determine the total dosage (mg). The inhaled dosage (μg / kg) was calculated by dividing the total dosage (μg) by the body weight of the animal (kg).

The dosage of Val8-GLP-1 deposited in the lungs was estimated as follows: inhaled dosage (μg / kg) was multiplied by 20 percent to produce the dosage of the estimated deposited lung (μg / kg). Aerosols with an MMEAD in the range of 1-2 μg MMAD have been shown to deposit in the lung with approximately 20 percent efficiency (Schlesinger RB, 1985. Comparative deposition of inhaled aerosols in experimental and human animals: a review JToxicol Environ Health 15: 197-214).

Pulmonary function All animals were weighed on days -5, 0 and 7. The breathing patterns (volume of lung ventilation, respiration rate and volume / minute) were monitored using a 0 'size pneumotachograph connected to a port in the head dome. The signals were collected on a personal computer using the Buxco XA Data Acquisition System (Buxco Electronics, Inc., Sharon, CT). At least 15 minutes of pre-exposure, the results were collected before the exposures began, followed by the collection of results through all exposure periods of 15 minutes. All results were analyzed as averages of 5 minutes. 20 Broncoa 1 veslar wash Bronchoalveolar lavage (BAL) was performed approximately 4 hours after each regimen of dosage. The animals were anesthetized with a Intravenous injection of 2% Brevital before BAL procedure, - Bronchoalveolar lavage was performed using a pediatric fiber optic bronchoscope (Olympus, model BF, Type 3C10, Lake Success, NY). The ronchoscope tip was weighed in a 5th to 7th generation duct of a lower lobe. The BALs alternated between the lobes of the right and left lung. Two 10 mL aliquots were infused and gently suctioned. The aliquots of the recovered wash fluid were used to determine leukocyte counts, red blood cell counts, leukocyte differentials, total proteins and lactate dehydrogenase.

Cellular Counts and Differentials A complete blood count was made in the non-concentrated BAL fluid using a Technicon Hl system (Technicon Instruments Corporation). Cell differentials for BAL were made by microscopic evaluation of 200 Wright stained cells.

Blood collection For the analysis of plasma pharmacokinetics of Val8-GLP-1, approximately 2 to 3 mL of blood were collected in tubes of vacuum vessel (vacutainer) EDTA of the cephalic or jugular vein before exposure and to 0.25, 0.5, 1, 2, 3, 4, 6 , 8 and 12 hours 5 of pre-exposure. To obtain the plasma, each tube was centrifuged at approximately 3000 rpm for 10 minutes at 10'C. The plasma samples were stored at -70'C until they were sent for the test.

Exposure concentration / Particle size and lung dosage The concentration of the average exposure for each dog exposed to Val8-GLP-1 was found in the range of 64.0 to 101.3 μg / L. The mean concentration (± SD) for all animals was 77.2 ± 16.9 μg / L. The particle size was measured as an average mass median aerodynamic diameter (MMEAD) of 0.91 μm with a geometric standard deviation (GSD) of 20 2.37.

The average dosage of Val8-GLP-1 deposited in the lungs of 6 dogs treated with Val8-GLP-1 was calculated as 240 ± 42 μg / kg (mean ± SD). The average inhaled dosage (which enters the tract v ijí 'r --l? d * A * aß *. ~ *. ^ A.ri ^ * ^ ^ ^ ^ ^ a ... 'AÉ respiratory that omits the deposition) was 1198 ± 208 μg / kg (mean ± SD). The results of the individual animals are shown in the following table.

Lung dosage estimated for animals exposed to Val8-GLP-1 aerosols Volume Conc / minute Number Dosing Average aerosol dose (ml) to the lung to the animal lung (μg / L) inhaled deposited (μg / kg) (μg / kg) 27682 0.07717 * 8406 1060 212 27684 0.07717 * 9563 1030 206 27685 0.10133 7260 1350 270 27686 0.06400 12733 1360 272 27687 0.06733 9214 950 190 27689 0.07600 11890 1400 288 * The concentration of the aerosol was not determined for animals 27682 and 27684, therefore, the average concentration of aerosol was used to calculate a dosage to the inhaled lung estimated (μg / kg). m - ^ - ^ rs ^^^? -M ^^ Á .. 'No significant changes in body weight were observed during the treatment phase. The initial (Day -5) and final (Day 7) body weights were 9.5 ± 0.9 (mean ± SD, n = 6) and 9.7 ± 0.9 kg, respectively. No marked changes in respiration frequency were observed. Slight increases in the volume of lung ventilation and volume / minute were measured, but the results were potentially highly variable due to the short acclimation period before the initiation study. The results of individual animals are shown in the following table.

Changes in lung function during exposure to Val8-GLP-1 Animal Volume Volume Frequency / minute respiration lung ventilation (bpm) (mL / m? N) (mL) Number 10 '15' 10 '15' 10 '15' 27682 119.0 152.1 105.1 74 16.0 95.7 7265 9360 8592 27684 104.6 99.4 100.8 119.2 118.2 108.7 10300 9117 9272 27685 165.9 178.5 209.6 58.1 49.4 42.5 7938 7465 6377 27686 325.1 586.2 374.4 56.3 33. 59.4 12200 13300 12700 B Animal Volume Volume Volume / minute respiration lung ventilation (bpm) (mL / rnin) (mL) Number 10 '15' 10 '15' 10 '15' 27689 184.5 367.6 454.0 39.8 46.8 33.3 7271 13400 15000 Average 190.8 267.6 237.8 64.6 63.3 65.0 9044 10369 10120 Stdev 82.8 180.7 145.3 29, 32.2 30.3 1956 2426.1 3141 Average 239.0 216.8 207.6 45.9 51.4 64.4 ** 8448 8744.8 8223 ** (Base line) Stdev 161.4 137.2 209.5 12 10.2 32.3 2614 2614 156 ( (Base line) * minutes during exposure (values represent the average in 5 minutes) ** average calculated from n = 5 (instead of n = 6) because no value was recorded for animal # 27689 a 15-minute exposure.

Comparison of pharmacokinetics after subcutaneous and pulmonary administration of Val8-GLP-1 C max route (h) AUC0_tt (1) Tl / 2 (a) release (ng / mL) (ng * h / mL) (hours) Subcutaneous 10.53 ± 1.06 0.7110.14 36.37 ± 2.18 1.26 ± 0.11 Inhalation 8.66 ± 0.90 1.54 ± 0.59 35.2115.91 1.1910.11 * All values are reported as mean ± SEM (standard error of the mean). (l) AUC0_t, = area under the plasma concentration curve from time 0 to t, where t = 12 hours of post-dosing.

Plasma concentrations of immunoreactive Val8-GLP-1 (Table 3) were measured by a competitive radioimmunoassay (RIA). Absorption of Val8-GLP-1 via both release routes appeared to be rapid, reaching substantial plasma concentrations at 15 minutes post-dosing. The plasma time profiles were similar for subcutaneous injections and inhalation. The average Tmax value for inhalation was greater than that for subcutaneous injection. Also, the - ^^ ^ £ ^^^^^^^^^,. R ..., ._ ^ ........ - ^ ^ ^ ^^^^^^^^^^^^^^ ? ¿^ Ívjms¿. concentration of Val8-GLP-l », ¡_§ > High plasma (close to Cmax) achieved by inhalation appeared to remain near the level for a longer period of time than after subcutaneous injection.

Based on the average AUC values, the bioavailability of inhaled Val8-GLP-1 (inhaled dosage averaged 1198 μg / kg) in relation to subcutaneous injection (100 μg / kg) was approximately 7.7%. On the basis of the dosage of the deposited lung, it was estimated as 240 μg / kg, the bioavailability in relation to the subcutaneous injection was 40%.

LIST OF SEQUENCES < 110 > Eli Lilly and Company < 120 > METHOD FOR ADMINISTERING INSULINOTROPIC PEPTIDES < 130 > X-12013 < 140 > < 141 > < 150 > US 60/098273 < 151 > 1998-08-28 15 < 160 > 4 < 170 > Patenteln Ver. 2.0 < 210 > 1 < 211 > 29 < 212 > PRT < 213 > Artificial Sequence < 220 > : .. ~ ~ ik ~ ... '. ^.-A ^ - ^ a ^. ^. ^^ tr -. te J .. ^^? J- í?, ^ ,, ^ .. * ,, -... .m ?. -is ^ -A ', ^, ^ & ^ m ^^^. ^^?, ^^: il ^^^^ < 223 > Description of the Sequence Artificial Synthetic Sequence < 220 > < 223 > Xaa at position 1 is selected from the group consisting of Ala, Gly, Val, Thr, lie and alpha-methyl-Ala. < 220 > < 223 > Xaa at position 14 is selected from the group consisting of Glu, Gln, Ala, Thr, Ser and Gly. < 220 > < 223 > Xaa at position 20 is selected from the group consisting of Glu, Gln, Ala, Thr, Ser and Gly. < 400 > 1 Xaa Glu Gly Thr Phe Thr Ser Asp Val Ser Ser Tyr Leu Xaa Gly Gln 1 5 10 15 Ala Ala Lys Xaa Phe He Ala Trp Leu Val Lys Gly Arg 20 25 < 210 > 2 < 211 > 28 < 212 > PRT < 213 > Homo Sapiens < 220 > < 223 > Description of the Sequence Artificial Synthetic Sequence < 220 > < 223 > Xaa at position 28 is selected from the group consisting of Lys and Lys-Gly. < 400 > 2 His Wing Glu Gly Thr Phe Thr Being Asp Val Being Ser Tyr Leu Glu Gly 1 5 10 15 Gln Ala Ala Lys Glu Phe He Ala Trp Leu Val Xaa 20 25 < 210 > 3 < 211 > 29 < 212 > PRT < 213 > Artificial Sequence < 220 > * & w »- ^ JÍ» < 223 > Xaa at position 19 is selected from the group consisting of Lys and Arg. < 220 > < 223 > Description of the Artificial Sequence: synthetic sequence < 400 > 3 Wing Glu Gly Thr Phe Thr Being Asp Val Being Ser Tyr Leu Glu Gly Gln 1 5 10 15 Wing Ala Xaa Glu Phe He Wing Trp Leu Val Lys Gly Arg 20 25 < 210 > 4 < 211 > 31 < 212 > PRT < 213 > Artificial Sequence < 220 > < 223 > Description of the Artificial Sequence: synthetic or semi-synthetic sequence < 400 > 4 His Val Glu Gly Thr Phe Thr Ser Asp Val Ser Ser Tyr Leu Glu Gly 1 5 10 15 Gln Ala Ala Lys Glu Phe He Ala Trp Leu Val Lys Gly Arg Gly 20 25 30 - > \ ^? ^^^ Is it noted that in relation to this date, the best method known by the applicant to carry out the aforementioned invention is that which is clear from the present description of the invention.

Having described the invention as above, the content of the following is claimed as property. fifteen twenty

Claims (40)

1. The use of a glucagon-like peptide-1 (GLP-1) molecule for the preparation of a medicament for the treatment of hyperglycemia to deliver a GLP-1 molecule selected from the group consisting of GLP-1, GLP analogs -1 or GLP-1 derivatives to a patient in need thereof by pulmonary means.

2. The use according to claim 1 wherein the GLP-1 molecule is released to the patient's lower passage.

3. The use according to claim 2 wherein the GLP-1 molecule is deposited in the alveoli.

4. The use according to claim 1 wherein the GLP-1 molecule is inhaled through the mouth of the patient.

5. The use according to claim 1 wherein the GLP-1 molecule is administered as a pharmaceutical formulation comprising the molecule of GLP-1 in a pharmaceutically acceptable vehicle.

6. The use according to claim 5 wherein the formulation is selected from the group consisting of a solution in an aqueous medium and a suspension in a non-aqueous medium.

7. The use according to claim 6 wherein the formulation is administered as an aerosol.

8. The use according to claim 5 wherein the formulation is in the form of a dry powder

9. The use according to claim 5 wherein the GLP-1 molecule has a particle size less than about 10 microns MMAD.

10. The use according to claim 9 wherein the GLP-1 molecule has a particle size of about 1 to about 5 microns MMAD.

11. The use according to claim 10 wherein the GLP-1 molecule has a size of gi | jta | A particle of approximately 2 to approximately 3 microns MMAD.

12. The use according to claim 1 wherein at least about 10% of the released GLP-1 molecule is deposited in the lung.

13. The use according to claim 1 wherein the GLP-1 molecule is released from an inhalation device suitable for pulmonary administration and capable of depositing the GLP-1 molecule in the lungs of the patient.

14. The use according to claim 13 wherein the device is selected from the group consisting of a nebulizer, a metered dose inhaler, a dry powder inhaler and an omitting device.

15. The use according to claim 14 wherein the device is a dry powder inhaler.

16. The use according to claim 1 wherein the GLP-1 molecule is selected from the group consisting of the GLP-1 analogs and derivatives of JM «Mj fis? ííí .- 'is aiL s aaaí' HHF ifí m * m ß *? GLP-1

17. The use according to claim 16 wherein the GLP-1 molecule is a GLP-1 analogue.

18. The use according to claim 17 wherein the GLP-1 analog is selected from the group consisting of Val8-GLP-I (7-37) OH, Gly8-GLP-I (7-37) OH and Asp8-GLP -1 (7-37) OH.

19. The use according to claim 18 wherein the GLP-1 analog is Val8-GLP-1 (7 -37) OH

20. The use according to claim 18 wherein the GLP-1 analog is Gl and 8-GLP-1 (7-37) OH

21. The use of an effective dosage for the preparation of a medicament for the treatment of hyperglycemia of a GLP-1 molecule to a patient in need thereof by pulmonary release.

22. The use according to claim 21 wherein the GLP-1 molecule is administered as a pharmaceutical formulation comprising the molecule of GLP-1 in a pharmaceutically acceptable vehicle

23. The use according to claim 21 wherein the GLP-1 molecule is Val8-GLP-1 (7-37) OH

24. The use according to claim 21 wherein the GLP-1 molecule is Gly8-GLP-1 (7 -37) OH.

25. The use according to claim 21 10 wherein the GLP-1 molecule is released from an inhalation device suitable for pulmonary administration and capable of depositing the GLP-1 molecule in the lungs of the patient.

26. The use according to claim 25 wherein the device is an atomizer or a dry powder inhaler.

27. The use according to claim 25 20 wherein a drive of the device delivers about 40 μg to about 4,000 μg of a GLP-1 molecule.

28. The use according to claim 25 25 wherein a drive of the device administers about 80 μg to about 2,000 μg of a GLP-1 molecule.

29. The use according to claim 25 wherein a drive of the device delivers about 160 μg to about 1,000 μg of a GLP-1 molecule.

30. The use according to claim 25 wherein a drive of the device delivers about 320 μg to about 500 μg of a GLP-1 molecule.

31. The use of a medicament for treating hyperglycemia wherein an effective dose of a GLP-1 molecule is administered to a patient in need thereof by pulmonary means.

32. The use according to claim 31 wherein the GLP-1 molecule is administered as a pharmaceutical formulation comprising the GLP-1 molecule in a pharmaceutically acceptable carrier.

33. The use according to claim 31 wherein the GLP-1 molecule is Val8-GLP-1 (7-37) OH.

34. The use according to claim 31 wherein the GLP-1 molecule is Gly8-GLP-1 (7 -37) OH

35. The use according to claim 31 5 wherein the GLP-1 molecule is released from an appropriate inhalation device for pulmonary administration and capable of depositing the GLP-1 molecule in the lungs of the patient.

36. The use according to claim 35 wherein the device is selected from the group consisting of an atomizer and a dry powder inhaler.

37. The use according to claim 35 wherein a drive of the device delivers about 40 μg to about 4,000 μg of the GLP-1 molecule.

38. The use according to claim 35 wherein a drive of the device delivers about 80 μg to about 2,000 μg of the GLP-1 molecule.

39. The use according to claim 35 wherein a drive of the device delivers about 160 μg to about 1,000 μg of the GLP-1 molecule.

40. The use according to claim 35 wherein a drive of the device delivers about 320 μg to about 500 μg of the GLP-1 molecule. SUMMARY OF THE INVENTION The claimed invention relates to a method for administering glucagon-like peptide-1 molecules by inhalation, a method for treating diabetes by administering glucagon-like peptide-1 molecules by inhalation and a method of treating hyperglycemia, administering peptide molecules. 1 similar to glucagon by inhalation. ggga ^^ S ^? tít ^ í