MXPA06008394A

MXPA06008394A - The combined use of glp-1 agonists and gastrin for regulating blood glucose levels

Info

Publication number: MXPA06008394A
Application number: MXPA/A/2006/008394A
Authority: MX
Inventors: Cruz Antonio; Pastrak Aleksandra; Hew Yin
Original assignee: Cruz Antonio; Hew Yin; Pastrak Aleksandra; Waratah Pharmaceuticals Inc
Priority date: 2004-01-30
Filing date: 2006-07-25
Publication date: 2007-04-10

Abstract

The invention relates to compositions, conjugates, and methods for the prevention and/or treatment of a condition and/or disease comprising a therapeutically effective amount of a GLP-1 agonist and a gastrin compound. The combination of a GLP-1 agonist and a gastrin compound provides beneficial effects, in particular sustained beneficial effects, in the prevention and/or treatment of conditions and/or diseases for which either a GLP-1 agonist or a gastrin compound have been demonstrated to have a therapeutic effect, including but not limited to diabetes, hypertension, chronic heart failure, fluid retentive states, obesity, metabolic syndrome and related diseases and disorders. Combinations of a GLP-1 agonist and a gastrin compound can be selected to provide unexpectedly additive effects or synergistic effects.

Description

ZW), Eurasian (AM, AZ, BY, KG, KZ, MD, RU, TJ, TM), For two-letter codes and other abbreviations. He referred to "GuidEuropean (AT, BE, BG, CH, CY, CZ, DE, DK, EE, ES, Fl, ance Notes on Codes and Abbreviations" appearing at the beginning- FR, GB, GR, HU, IB, IS, IT, LT, LU, MC, NL, PL, PT, RO, no ofeach regular issue of the PCT Gazette SE, SI, SK, TR), OAPI (BF, BJ, CF, CG, Cl, CM, GA, GN , GQ, GW, ML, MR, NE, SN, TD, TG). Published: - without inlemational search report and what is republished upon receipt of that report THE COMBINED USE OF GLUCOSE (GLP-1) AND GASTRINE-LIKE AGONISTS 1 SIMILAR TO REGULATE BLOOD GLUCOSE LEVELS Field of the Invention The invention relates generally to compositions, conjugates and methods comprising a GLP-1 agonist and a gastrin compound and use thereof. Background of the Invention Glucose-like peptide-1 (GLP-1) is a physiological incretin hormone of the lower gastrointestinal tract. GLP-1 has important physiological activities that include the stimulation of glucose-dependent insulin secretion, the inhibition of glucagon secretion and gastric emptying, inhibition of dietary intake, intensification of glucose use, preservation of beta cells, inhibition of apoptosis by beta cells, and induction of beta cell proliferation, [see, Nauck, MA Acta Diabetol, 1998, 35: 117-129; Holst J.J. Diabetes Metab Res Rev 2002, 18: 430-441; Reimer, R.A. et al, Endocrinology 142 (10): 4522-4528; Drucker, D.J., Molecular Endocrinology, 2003, 17 (2) 161-171 and http: // www. qlucagon. com for GLP-1 reviews]. The previously established activities of GLP-1, make it a highly desirable therapeutic agent for the treatment of many conditions and diseases that REF: 174657 they include diabetes, obesity, gastric ulcers, hypertension, chronic heart failure, fluid retention states, metabolic syndrome and related diseases and disorders. The citation of any reference herein is not an admission that such a reference is available as a prior art for the current invention.

Brief Description of the Invention The combination of the GLP-1 agonist and a gastrin compound provides beneficial effects in the prevention and / or treatment of conditions and / or diseases for which either a GLP-1 agonist or a gastrin compound, has been shown to have a therapeutic effect, including but not limited to diabetes, hypertension, chronic heart failure, fluid retention states, obesity, metabolic syndrome, and related diseases and disorders. The combinations of a GLP-1 agonist and a gastrin compound can be selected to provide unexpectedly additive or greater effects than the additive effects, that is, synergistic effects. A composition, conjugate or method comprising a GLP-1 agonist and a gastrin compound that employs different mechanisms to achieve maximum therapeutic efficacy, can improve tolerance to therapy with a reduced risk of side effects that may result from higher doses or monotherapies of a longer period (ie, therapies with each compound alone). A composition, conjugate or method of the invention will allow the use of lower doses of one or both compounds with reduced adverse toxic effects of each compound. A suboptimal dose can provide an increased safety margin and can also reduce the cost of a drug necessary to achieve prophylaxis and therapy. In certain aspects of the invention, the increasing convenience of a simple combination dosing unit can result in improved compliance. Other advantages of a composition, conjugate or combination therapy may include greater stability towards degradation and metabolism, longer duration of action and / or longer duration of action or effectiveness at particularly low doses. Established widely, the invention relates to conjugated compositions and methods for the prevention and / or treatment of a condition and / or disease comprising a therapeutically effective amount of a GLP-1 agonist and a gastrin compound that provides beneficial effects. A composition, conjugate or method of the invention can provide sustained beneficial effects following treatment or termination of treatment. Prolonged efficacy may be evident by increased production of C-peptide, increases in pancreatic insulin production and / or around normal glucose levels in blood compared to GLP-1 or gastrin alone. In one aspect, the invention contemplates a composition, preferably a pharmaceutical composition, comprising a GLP-1 agonist and a gastrin compound that provides beneficial effects relative to each compound alone. In another aspect the invention provides a pharmaceutical composition comprising a GLP-1 agonist and a gastrin compound that provides beneficial effects, preferably sustained beneficial effects after treatment. A pharmaceutical composition may optionally comprise a pharmaceutically acceptable excipient carrier or vehicle. The invention also contemplates a separate container pharmaceutical composition which is intended for simultaneous or sequential administration to provide beneficial effects, preferably sustained beneficial effects comprising a GLP-1 agonist and a gastrin compound both optionally together with excipient carriers or carriers pharmaceutically acceptable The invention further contemplates a conjugate comprising a GLP-1 agonist that interacts with or is linked to a gastrin compound to provide beneficial effects, preferably sustained beneficial effects discussed herein.

The invention still further contemplates methods for the preparation of compositions and conjugates of the invention which result in compositions and conjugates with beneficial effects preferably sustained beneficial effects. In one aspect of the invention, a method is provided for the preparation of a stable pharmaceutical composition of a GLP-1 agonist and a gastrin compound, adapted to provide beneficial effects, preferably beneficial effects "sustained after treatment, comprising preparing a composition comprising the GLP-1 agonist a gastrin compound and a pharmaceutically acceptable carrier, excipient or carrier effective to physically stabilize the GLP-1 agonist In another aspect of the invention, a method for the preparation of a pharmaceutical composition is provided of a GLP-1 agonist comprising mixing a GLP-1 agonist, a gastrin compound and a pharmaceutically acceptable carrier, excipient or carrier effective to physically stabilize the GLP-1 agonist and adapted to deliver beneficial effects, preferably sustained beneficial effects. The invention relates to a combination treatment n to prevent and / or treat a condition and / or disease discussed herein in a subject comprising administering to a subject a therapeutically effective amount of at least one GLP-1 agonist and a gastrin compound to provide beneficial effects. In one aspect of the invention, it provides a combination or intervention treatment which provides sustained beneficial effects after treatment. The invention further relates to the use of a GLP-1 agonist and a gastrin compound, a composition or conjugate of the invention for preventing and / or improving the severity of the disease, symptoms of the disease and / or periodicity of recurrences of a condition and / or illness- described here. The invention still relates to the prevention and / or treatment in a subject of diseases and / or conditions using a GLP-1 agonist, a gastrin and a gastrin compound, a composition or conjugate of the invention. In one aspect the invention provides a method for the prevention and / or intervention of a condition and / or disease discussed herein in a subject comprising administering at least one GLP-1 agonist and at least one gastrin compound or a composition or conjugate of the invention. A GLP-1 agonist and a compositional or conjugated gastrin compound can be administered directly to a subject or contacting the cells (e.g., stem cells or progenitor cells) and administered to a subject. In another aspect, the invention provides a method for the prevention and / or intervention of a condition and / or disease discussed herein in a subject comprising the administration of at least one GLP-1 agonist and at least one gastrin compound to a subject in need thereof to provide beneficial effects. In another aspect, the invention provides a method for the prevention and / or intervention of a condition and / or disease discussed herein in a subject, comprising coadministering at least one GLP-1 agonist and at least one gastrin compound to a subject that need it In a particular aspect, the invention relates to the induction of islet neogenesis in a subject comprising contacting the islet precursor cells with a GLP-1 agonist and a composition or conjugated gastrin compound of the invention in a sufficient amount to increase the proliferation of islet precursor cells in the subject thereby inducing islet neogenesis. In another aspect, the invention relates to a method for the treatment of diabetes mellitus in a patient in need thereof, by administering a gastrin compound and a GLP-1 agonist or a composition comprising a gastrin compound and a GLP-len agonist is an amount sufficient to effect the differentiation of pancreatic islet precursor cells from the patient into a mature insulin secretory cell and / or to stimulate the synthesis of insulin in existing islet cells.

The invention provides methods for the treatment of cells using a GLP-1 agonist and a gastrin compound of the invention, or compositions or conjugates of the invention. In particular, the invention relates to a method for expanding and differentiating stem cells or progenitor cells in insulin secreting cells, enhancing the proliferation of insulin secreting cells and / or supporting islet cells or precursor cells. The cells can be contacted with a GLP-1 agonist and a gastrin compound in culture or in a subject. In one aspect, there is provided a method for the treatment of a condition and / or disease comprising administering a GLP-1 agonist and a gastrin compound, a composition or conjugate of the invention with a plurality of cells to a subject in need of same to thereby produce beneficial effects, preferably sustained beneficial effects. In one embodiment, the conjugated compound compositions are administered systemically. In another aspect, the invention provides a method for the treatment of a subject with a condition and / or disease discussed herein comprising contacting a plurality of cells ex vivo with a GLP-1 agonist of a gastrin compound or a composition or conjugate of the invention, optionally culturing the cells and administering the cells to the subject in need thereof.

Also provided in particular aspects of the invention are methods and compositions for the treatment of diabetes in a patient in need thereof to implant in a diabetic patient pancreatic islet cells which have been exposed in culture to a sufficient amount of a gastrin compound. and a GLP-1 agonist or a composition or conjugate of the invention to increase the number of β-pancreatic cells in the islets, optionally the population of β-pancreatic cells can grow in culture with a sufficient time to expand the population of beta cells before the transplant. The invention also contemplates the use of a composition comprising a combination of at least one GLP-1 agonist and at least one gastrin compound for the preparation of one or more drugs for the prevention and / or treatment of a condition or disease. The invention further contemplates the use of a GLP-1 agonist in combination with a gastrin compound for the manufacture of a medicament for the treatment of a condition and / or disease. Still further, the invention provides the use of a GLP-1 agonist for the manufacture of a medicament for the treatment of a condition and / or disease to be used in combination with a gastrin compound. In one aspect, the invention relates to the use of synergistically effective amounts of at least one GLP-1 agonist and at least one gastrin compound for the preparation of a medication for the prevention or treatment of a condition and / or disease. In another aspect, the invention relates to the use of a GLP-1 agonist and a gastrin compound for the preparation of a medicament having a dilated profile of action relative to GLP-1 (7-37) or • exendin. The invention further provides the use of a pharmaceutical composition and a conjugate of the invention in the preparation of medicaments for the prevention and / or treatment of conditions and / or diseases. The medications provide beneficial effects preferably beneficial effects sustained after treatment. Since the present invention relates to a method for prevention and / or treatment comprising a combination of active agents which can be administered separately or as a conjugate, the invention also provides a kit comprising a GLP-1 agonist and a gastrin compound and a pharmaceutical composition or conjugate of the invention in the form of a kit. These and other aspects, features and advantages of the present invention will be apparent to those skilled in the art from the following detailed description.

Brief Description of the Figures The invention will be better understood with reference to the figures, which: Figure 1 is a graph showing the effect of islet neogenesis therapy using a GLP-1 agonist and a gastrin compound (INT-2) on fasting blood glucose in acutely diabetic NOD mice. The figure illustrates that the combination of GLP-1 (300μg / kg) treatment with gastrin (3μg / kg) reverses hyperglycemia and prevents death in NOD mice. Figure 2 are graphs showing individual fasting blood glucose levels in acutely diabetic NOD mice treated with the combination of GLP-1 (300 μg / kg) and gastrin (3 μg / kg). Figure 3 is a graph showing that the combination treatment of GLP-1 and gastrin re-establishes panc-septic insulin levels in a NOD mouse model. Figure 4 is a graph showing the correlation between pancreatic insulin content and fasting blood glucose levels. Figure 5 shows cells stained with insulin in acute diabetic NOD mice treated with vehicle and GLP-1 and gastrin. Figure 6 shows islet cell staining from the pancreatic duct in NOD mice treated with vehicle and GLP-1 and gastrin.

Glossary The mention of the numerical ranges by endpoints in the present, includes all the numbers and reactions proposed within the range (for example, from 1 to 5 includes 1, 1.5, 2, 2.75, 3, 3.90, 4 and 5). It will also be understood that all numbers and fractions thereof are presumed to be modified by the term around. In addition, it is understood that "a", "some" and "the" include plural referents unless the content clearly dictates otherwise. Thus, for example, reference to a composition containing a compound includes a mixture of two or more compounds. The term "around" means more or less 0.1 to 50%, or 10-40%, preferably 10-20%, more preferably 10% or 15%, of the number to which reference is made. The selected compounds described herein contain one or more asymmetric centers and can give rise to diastereomeric enantiomers and other stereoisomeric forms which can be defined in terms of absolute stereochemistry as (R) or (S). Therefore, the invention includes all such possible diastereomers and enantiomers as well as their racemic and optically pure forms. The optically active isomers (R) - and (S) - can be prepared by using chiral synthons or chiral reagents or resolved using conventional techniques. When the compounds described here contain centers of geometric asymmetry, and unless otherwise specified, the compounds are intended to include both geometric isomers E and A. All tautomeric forms are intended to be included within the scope of the invention. The terms "subject""individual" or "patient" refers to an animal that includes a warm-blooded animal such as a mammal which suffers or is suspected of having or is predisposed to a condition and / or disease as described herein. Preferably, the terms refer to a human. The terms also include crossed domestic animals for food, sport or as pets including; horses, cows, sheep, poultry, fish, pigs, cats, dogs and zoo animals. The methods herein for use in subjects / individuals / patients contemplate prophylactic as well as curative use. Typical subjects for treatment include persons susceptible to, suffering from or who have suffered a condition and / or disease discussed herein. The term "pharmaceutically acceptable carrier, excipient or vehicle" refers to a medium which does not interfere with the effectiveness or activity of an active ingredient and which is not toxic to the hosts to which it is administered. A carrier, excipient or carrier includes diluents, binders, adhesives, lubricants, disintegrants, bulking agents, emulsifying agents or "humectants, pH buffering agents, various materials such as absorbents that may be needed in order to prepare a particular composition." The use of such media and agents for "an active substance is well known in the art. In certain aspects of the invention an excipient carrier or vehicle carrier is selected to stabilize a gansite of GLP-1. The pharmaceutically acceptable salts include salts of acidic or basic groups which may be present in the compounds suitable for use in the present invention. Examples of pharmaceutically acceptable salts include sodium, calcium and potassium salts of carboxylic acid groups and salts of amino group chlorohydrates. Other pharmaceutically acceptable salts of the amino groups are bromohydrate, sulfate, acid sulfate, phosphate, acid phosphate, diacid phosphate, acetate, succinate; citrate, tartrate, lactate, mandelate, methanesulfonate, (mesylate) and salts of p-toluenesulfonate (tosylate). The terms "prevention and / or treatment", "prevention and / or treatment" or "prevention and / or intervention", refer to the administration to a subject of biologically active agents either before or after the onset of a condition and / or disease. A treatment can be carried out in an acute or chronic way. In particular, prevention includes the administration and care of a subject at risk of develop a condition and / or disease discussed here prior to the clinical onset of the condition and / or disease. The treatment or intervention refers to the administration and care of a subject in the diagnosis or later. An objective of prevention, treatment or intervention is to combat the condition or disease and includes the administration of the active compounds to prevent or delay the onset of symptoms or complications or alleviate the symptoms or complications or eliminate or partially eliminate the condition and / or disease. A beneficial effect refers to an effect of a combination of a GLP-1 agonist and a gastrin compound or a non-conjugated composition thereof, which is greater than the effect of any of the compounds alone. The beneficial effect includes favorable pharmacological and / or therapeutic effects and improved pharmacokinetic properties and biological activity. A beneficial effect can be an additive effect or a synergistic effect. In preferred embodiments of the invention, the beneficial effects include but are not limited to the following: reduced or absent inflammation of islets, decreased progression of the disease, increased survival or partial elimination of a condition and / or disease. In a particularly preferred embodiment, the beneficial effect is a sustained beneficial effect wherein the beneficial effect is sustained for a prolonged period of time. time after the termination of treatment. In one embodiment, one or more of the aforementioned effects is sustained for a prolonged period of time after the termination of the treatment. A beneficial effect can be sustained for at least about 2, 4, 6, 8, 10, 2 to 4 weeks, 2 to 6 weeks, 2 to 8 weeks, 2 to 12 weeks, 2 to 24 weeks, 2 weeks to 12 months, and 2 weeks to 18 months after treatment. The period of time that a beneficial effect is sustained can be correlated with the duration and schedule of the treatment. A subject can be treated continuously for about 2 to 8 weeks, 2 to 12 weeks, 2 to 16 weeks, 2 weeks to 6 months, 2 weeks to 12 months, or periodically. A sustained beneficial effect may be manifested as one or more increased production of C-peptide, increased production of pancreatic insulin and / or low or near normal blood glucose levels for a prolonged period after treatment. The beneficial effect can be a statistically important effect in terms of the statistical analysis of an effect of the two compounds against the effects of each of the compounds. "Statistically significant" or "statistically different" effects or levels with the two compounds compared to each compound alone may represent levels that are higher or lower than the standard In embodiments of the invention the difference may be 1.5, 2, 3, 4, 5 or 6 times higher or lower compared to the effect obtained with each compound alone. An "additive effect" of the GLP-1 agonist and a gastrin compound refers to a compound that is equal to the sum of the effects of the two individual compounds. A "synergistic effect" of a GLP-1 agonist and a gastrin compound refers to an effect that is greater than the additive effect which results from the sum of the effects of the two individual compounds. "Combination therapy", "combination therapy" and "administration in combination", are used interchangeably herein and means that the active ingredients are administered in parallel to a patient being treated. When administered in combination each component can be administered at the same time or sequentially in any order at a different time point. Therefore, each component can be administered separately but sufficiently close in time to provide the desired effect, in particular, an additive or synergistic beneficial effect. The first compound can be administered in a regimen which further comprises the treatment with the second compound. In certain embodiments, the term refers to the administration of a GLP-1 agonist and a gastrin compound to a patient within one year, including administration separately from two drugs each contains one of the compounds, as well as the simultaneous administration whether the two compounds combine or not in a formulation or whether they are two "separate formulations." A "medicament" refers to a composition pharmaceutical composition suitable for administering a pharmaceutically active compound (eg, a GLP-1 agonist or a gastrin compound) to a patient. "Therapeutically effective amount" "refers to the amount or dose of the active compounds (eg. example, GLP-1 agonist and gastrin compound), compositions or conjugates of the invention which will lead to one or more desired beneficial effects, preferably one or more sustained beneficial effects.A therapeutically effective amount can provide a dose which is sufficient for the prevention and / or treatment of a subject to be effective compared to no treatment. "Synergistically effective amount "refers" to the amount of doses of active compounds (eg, GLP-1 agonist and gastrin compound), compositions or conjugates of the invention that will provide a synergistic effect, in particular a beneficial synergistic effect. "Suboptimal dose or suboptimal dosage" refers to a dose or dosage of an active compound that is less than the optimal dose or dosage for that compound when used in monotherapy. The terms "associated", "bound", "interact", "interaction", or "interact" refer to any physical association "between molecules." The terms preferably refer to a stable association between two molecules due to, for example, electrostatic, hydrophobic, ionic, hydrogen-bound interactions, or covalent interactions Certain molecules that interact or associate interact only after one or more of them have been activated In the present context, "a GLP-1 agonist" is understood to refer to any compound, which includes peptide and non-peptide compounds, that fully or partially activate the human GLP-1 receptor In a preferred embodiment, the "GLP-1 agonist" is any small molecule of peptide or non-peptide that binds to a GLP-1 receptor, preferably with a constant affinity (D) or a power (EC50) below 1 μM, for example, below 100 nM as a measure for methods known in the art. ca (see for example, WO 98/08871) and exhibits insulinotropic activity, wherein the insulinotropic activity can be measured in in vivo or in vitro assays known to those of ordinary skill in the art. For example, the GLP-1 agonist It can be administered to an animal and the measured insulin concentration over time. In one embodiment, the GLP-1 agonist is selected from the group consisting of GLP-1 (7-36) -amide, GLP-1 (7-37), a GLP-1 (7-36) -amide analog, a GLP-1 analog (7-37), or a derivative of any of these. In the current application, the designation "an analog" is used to designate a peptide wherein one or more amino acid residues of the parent peptide have been replaced by another amino acid residue and / or one or more amino acid residues of the parent peptide. have been removed and / or where one or more amino acid residues have been added to the precursor peptide. Such addition can occur either at the N-terminal end or the C-terminal end of the precursor peptide or both. Typically "an analogue" is a peptide wherein 6 or fewer amino acids have been substituted and / or added and / or removed from the precursor peptide, more preferably a peptide wherein 3 or fewer amino acids have been substituted and / or added and / or deleted of the precursor peptide, and more preferably, a peptide wherein an amino acid has been substituted and / or added and / or removed from the precursor peptide. In the current application, "a derivative" is used to designate a peptide or analog thereof that is chemically modified by introducing for example, ester, alkyl or lipophilic functionalities in one or more amino acid residues of the peptide or analogue thereof. Methods for identifying GLP-1 agonists are described in WO 93/19175 (Novo Nordisk A / S) and examples of appropriate GLP-1 analogs and derivatives that can be used in accordance with the present invention include those which are referred to in WO 99. / 43705 (Novo Nordisk A / S), WÓ 99/43706 (Novo Nordisk A / S), WO 99/43707 (Novo Nordisk A / S), WO 98/08871 (Novo Nordisk A / S), WO 99/43708 (Novo Nordisk A / S), WO 99/43341 (Novo Nordisk-A / S), WO 87/06941 (The General Hospital Corporation), WO 90/11296 (The General Hospital Corporation), WO 91/11457 (Buckley et al.), WO 98/43658 (Eli .Lilly & Co.), EP 0708179-A2 (Eli Lilly &Co.), EP 0699686-A2 (Eli 'Lilly- &Co.), WO 01/98331 (Eli Lilly &Co.). In one embodiment, the GLP-1 agonist is a derivative of GLP-1 (7-36) -amide, GLP-1 (7-37), a GLP-1 (7-36) -amide analog or a GLP-1 (7-37) analog, comprising a lipophilic substituent. In this embodiment of the invention, the GLP-1 derivative preferably has three lipophilic substituents, more preferably two lipophilic substituents, and more preferably a lipophilic substituent linked to the precursor peptide (ie GLP-1 (7-36) -amide, GLP- 1 (7-37), a GLP-1 (7-36) -amide analog or a GLP-1 (7-37) analog, where each lipophilic substituent preferably has 4- 40 carbon atoms, more preferably 8-30 carbon atoms, still more preferably 8-25 carbon atoms, even more preferably 12-25 carbon atoms, and more preferably 14-18 carbon atoms. In one embodiment, the lipophilic substituent comprises a partially or fully hydrogenated cyclopentanphenatrene skeleton. In another embodiment, the lipophilic substituent is a straight or branched chain alkyl group. In yet another embodiment, the lipophilic substituent is a. acyl group of a straight or branched chain fatty acid. Preferably, the lipophilic substituent is an acyl group having the formula CH3 (CH2) nC0-, wherein. n is an integer from 4 to 38, preferably an integer from 12 to 38, and more preferably CH3 (CH2) 12C0-, CH3 (CH2)? 4CO-, CH3 (CH2) 16C0-, CH3 (CH2)? 8CO-, CH3 (CH2) 20CO- and CH3 (CH2) 22CO-. In a more preferred embodiment, the lipophilic substituent is tetradecanoyl. In a more preferred embodiment, the lipophilic substituent is hexadecanoyl. In a further embodiment of the present invention, the lipophilic substituent has a negatively charged group such as a carboxylic acid group. For example, the lipophilic substituent can be an acyl group of a straight or branched chain alkane, w-dicarboxylic acid of the formula HOOC (CH2) mCO-, wherein m is a whole from 4 to 38, preferably an integer from 12 to 38, and more preferably is HOOC (CH2)? 4CO-, HOOC (CH2)? SCO-, HOOC (CH2) a8CO-, HOOC (CH2) 20CO- or HOOC ( CH2) 22CO-.

In the GLP-1 derivatives of the invention, the lipophilic substituents contain a functional group which can be linked to one of the following functional groups of an amino acid of the precursor GLP-1 peptide: (a) the amine group bonded to the alpha carbon of the amino acid terminal N, (b) the carboxy group bonded to the alpha carbon of the C-terminal amino acid, (c) -the epsilon-amino group of any residue Lys, (d) the carboxy group of the R group of any residue Asp and Glu, (e) the hydroxy group of the group R of any residue Tyr, Ser and Thr, (f) the amino group of the group R of any residue Trp, Asn, Gln, Arg, and His, or (g) the thiol group of the group R of any Cys residue. In one embodiment, a lipophilic substituent is linked to the carboxy group of the R group of any Asp and Glu residue.

In another embodiment, a lipophilic substituent is bonded to the carboxy group bonded to the alpha carbon of the C-terminal amino acid.

In a more preferred embodiment, a lipophilic substituent is linked to the epsilon-amino group of any Lys residue. In a preferred embodiment of the invention, the lipophilic substituent is linked to the precursor GLP-1 peptide by means of a spacer. A spacer must contain at least two functional groups, one to bind to a functional group of the lipophilic substituent and the other to a functional group of the precursor GLP-1 peptide. In one embodiment, the spacer is an amino acid residue except Cys or Met, or a dipeptide such as Gly-Lys. For purposes of the present invention, the phrase "a dipeptide such as Gly-Lys" means any combination of two amino acids except Cys or Met, preferably a dipeptide wherein the C-terminal amino acid residue is Lys, His or Trp, preferably Lys, and the terminal amino acid residue N is Ala, Arg, Asp, Asn, Gly, Glu, Gln, Lie, Leu, Val, Phe, Pro, Ser, Tyr, Thr, Lys, His and Trp. Preferably, an amino group of the precursor peptide forms an amide bond with a carboxylic group of the amino acid residue or spacer dipeptide, and an amino group of the amino acid residue or dipeptide spacer forms an amide bond with a carboxyl group of the lipophilic substituent. Preferred spacers are lysyl, glutamyl, asparagyl, glycyl, beta-alanyl and gamma-aminobutanoyl, each one of which constitutes an individual modality. More preferred spacers are glutamyl and beta-alanyl. When the spacer is Lys, Glu or Asp, the carboxyl group thereof can form an amide bond with an amino group of the amino acid residue, and the amino group thereof can form an amide bond with a carboxyl group of the lipophilic substituent. When Lys is used as the spacer, an additional spacer may in some cases be inserted between the e-amino group of Lys and the lipophilic substituent. In one embodiment, such additional spacer is succinic acid which forms an amide bond with the e-amino group of Lys and with an amino group present in the lipophilic substituent. In another embodiment such additional spacer is Glu or Asp which forms an amide bond with the e-amino group of Lys and another amide bond with a carboxylic group present in the lipophilic substituent, that is, the lipophilic substituent is a residue of acylated Ne lysine. . In another embodiment, the spacer is an unbranched alkane, w-dicarboxylic acid group having from 1 to 7 methylene groups, which spacer forms a bridge between an amino group of the precursor peptide and an amino group of the lipophilic substituent. Preferably, the spacer is succinic acid. In a further embodiment, the lipophilic substituent with the linked spacer is a group of the formula CH3 (CH2) pNH-CO (CH2) qC0-, where p is an integer from 8 to 33, preferably from 12 to 28 and q is an integer from 1 to 6, preferably 2. In a further embodiment, the lipophilic substituent with the linked spacer is a group of the formula CH3 (CH2) rCO-NHCH (COOH) (CH2) 2CO-, wherein r is an integer from 4 to 24, preferably from 10 to 24. In a further embodiment, the lipophilic substituent with the linked spacer is a group of the formula CH3 (CH2) sCO-? HCH ((CH2) 2COOH) CO-, where s is an integer from 4 to 24 , preferably from 10 to 24. In a further embodiment, the lipophilic substituent is a group of the formula COOH (CH) tCO- where t is an integer from 6 to 24. In a further embodiment, the lipophilic substituent with the linked spacer is a group of the formula? HCH (COOH) (CH2) 4? H-CO (CH2) ICH3, where u is an integer from 8 to 18. In a further embodiment, the lipophilic substituent with the linked spacer is a group of the formula CH3 (CH2) vCO-? H- (CH2) Z-C0, where v is an integer from 4 to 24 and z is an integer from from 1 to 6. In a further embodiment, the lipophilic substituent with the linked spacer is a group of the formula - NHCH (COOH) (CH2) 4NH-COCH ((CH2) 2COOH) NH-CO (CH2) WCH3, wherein w is an integer from 10 to 16. In a further embodiment, the lipophilic substituent with the linked spacer is a group of the formula NHCH (COOH) (CH2) 4NH-CO (CH2) 2CH (COOH) NHCO (CH2) XCH3, wherein x is zero or an integer from 1 to 22, preferably 10 to 16. In yet another modality the GLP-1 agonist is Arg34, Lys26 (Ne- (? -Glu (Na-hexadecanoyl))) -GLP-1 (7-37). In yet another embodiment the GLP-1 agonist is selected from the group consisting of Gly8-GLP-1 (7-36) -amide, Gly8-GLP-1 (7-37), Val8-GLP-1 (7-36) -amide, Val8-GLP-1 (7-37), Val8Asp22-GLP-I (7-36) -amide, Val8Asp22-GLP-I (7-37), Val8Glu22-GLP-1 (7-36) -amide, Val8Glu22-GLP-I (7-37), Val8Lys22- GLP-l (7-36) -amide, Val8Lys22-GLP-l (7-37), Val8Arg22-GLP-l (7-36) -amide, Val8Arg22-GLP-l (7-37), Val8His 2-GLP -l (7-36) -amide, Val8Eis22-GLP-I (7-37), analogs thereof and derivatives of any of these. In yet another embodiment the GLP-1 agonist is selected from the group consisting of Arg2d-GLP-1 (7-37); Arg-GLP-1 (7-37); Lys36-GLP-1 (7-37); Arg26'34Lys36-GLP-I (7-37); Arg26 '34 -GLP -1 (7-37); Arg26'34Lys40-GLP-I (7-37); Arg26Lys36-GLP-1 (7-37); Arg34Lys36-GLP-I (7-37); Val8Arg22-GLP-I (7-37); M.et8Arg22-GL, P-1 (7-37); Gly8His22-GLP-I (7-37); Val8His22-GLP-I (7-37); Met8His22-GLP-I (7-37); His37-GLP-1 (7-37); Gly8-GLP-1 (7-37); Val8-GLP-1 (7-37); Met8-GLP-1 (7-37); Gly8Asp22-GLP-l (7-37); Val8Asp22-GLP-I (7-37); Met8Asp22-GLP-I (7-37); Gly8Glu22-GLP-I (7-37); Val8Glu22-GLP-I (7-37); Met8Glu22-GLP-l (7-37); Gly8Lys22-GLP-I (7-37); Val8Lys22-GLP-I (7-37); Met8Lys22-GLP-I (7-37); Gly8Arg22-GLP-I (7-37); Val8Lys22His37-GLP-I (7-37); Gly8Glu22His37-GLP-I (7-37) Val8Glu22His37-GLP-I (7-37); Met8Glu22His37-GLP-I (7-37) Gly8Lys22His37-GLP-I (7-37); Met8Lys 2His37-GLP-I (7-37) Gly8Arg22His37-GLP-I (7-37); Val8Arg22His37-GLP-I (7-37) Met8Arg22His37-GLP-I (7-37); Gly8His22His37-GLP-l (7-37) Val8His22His37-GLP-I (7-37); . Met8His22His37-GLP-I (7-37) Gly8His37-GLP-I (7-37); Val HIS37-GLP-I (7-37); Met8His37-GLP-I (7-37); Gly8Asp22 His37-GLP-1 (7-37); Val8Asp22His37-GLP-I (7-37); Met8Asp22His37-GLP-I (7-37); Arg26-GLP-1 (7-36) -amide; Arg34-GLP-1 (7-36) -amide; Lys36-GLP-1 (7-36) -amide; Arg26'34Lys36-GLP-I (7-36) -amide; Arg26 '34 -GLP -1 (7-36) -amide; Arg26 3 Lys 40 -GLP-I (7-36) -amide; Arg26Lys36-GLP-I (7-36) -amide; Arg34Lys36-GLP-I (7-36) -amide; Gly8-GLP-1 (7-36) -amide; Val8-GLP-1 (7-36) -amide; Mets-GLP-1 (7-36) -amide; Gly8Asp22-GLP-I (7-36) -amide; Gly8Glu22His37-GLP-I (7-36) -amide; Val8Asp2-GLP-1 (7-36) -amide; Met8Asp22-GLP-I (7-36) -amide; Gly8Glu22-GLP-I (7-36) -amide; Val8Glu22-GLP-1 (7-36) -amide; Met8Glu22-GLP-I (7-36) -amide; Gly8Lys22-GLP-I (7-36) -amide; Val8Lys22-GLP-I (7-36) -amide; Met8Lys22-GLP-I (7-36) -amide; Gly8His22His37-GLP-I (7-36) -amide; Gly8Arg22-GLP-I (7-36) -amide; Val8Arg22-GLP-l (7- 36) -amide; Met8Arg22-GLP-I (7-36) -amide; Gly8His22-GLP-I (7-36) -amide; 'Val8His22-GLP-1 (7-36) -amide; Met8His22-GLP-I (7-36) -amide; His37-GLP-1 (7-36) -amide; Val8Arg22His37-GLP-I (7-36) -amide; Met8Arg22His37-GLP-1 (7-36) -amide; Gly8His37-GLP-l _ (7-36) -amide; Val8His37-GLP-I (7-36) -amide; Met8His37-GLP-I (7-36) -amide; Gly8Asp 2His37-GLP-1 (7-36) -amide; Val8Asp22His37- GLP-1 (7-36) -amide; Met8Asp22His37-GLP-I (7-36) -amide; Val8Glu22His37-GLP-I (7-36) -amide, - Met8Glu22His37-GLP-I (7-36) -amide; Gly8Lys22His37-GLP-I (7-36) -amide; Val8Lys22His37-GLP-I (7-36) -amide; Met8Lys22His37-GLP-I (7-36) -amide; Gly8Arg22His37- - GLP-1 (7-36) -amide; Val8His22His37-GLP-I (7-36) -amide; Met8His2His37-GLP-I (7-36) -amide; and derivatives thereof. In yet another embodiment the GLP-1 agonist is selected from the group consisting of Val8Trp19Glu22-GLP-I (7-37), Val8Glu22Val25-GLP-I (7-37), Val8TyrlsGlu22-GLP-I (7-37), Val8TrplsGlu22 -GLP-I (7-37), Val8Leu16Glu22-GLP-I (7-37), Val8Tyr18Glu22-GLP-I (7-37), Val8Glu22His37-GLP-I (7-37), Val8Glu22Ile33-GLP-I (7-37), Val8Trp16Glu22Val25Ile33-GLP-I (7-37), Val8Trpa6Glu22Ile33-GLP-I (7-37), Val8Glu22Val25Ile33-GLP-I (7-37), Val8Trp16Glu22Val25-GLP-I ( 7-37), analogs thereof and derivatives of any of these. In yet another embodiment the GLP-1 agonist is a stable analog / derivative GLP-1. Through this application a "Stable GLP-1 analog / derivative" means an analogous GLP-1 or a. derived from a GLP-1 analog that exhibits half-life of one plasma elimination in vivo of at least 10 hours in man, as determined by the method described below. Examples of stable GLP-1 analog / derivatives can be found in WO 98/08871 and WO 99/43706. The method for determining the plasma elimination half-life of a compound in a man is: The compound is dissolved in an isotonic buffer, pH 7.4, PBS or any other suitable buffer solution. The dose is injected peripherally, preferably in the abdomen or thigh. Blood samples for the determination of active compound are taken at frequent intervals, and for a duration sufficient to cover the terminal elimination part (eg, Pre-dose, 1, 2, 3, 4, 5, 6, 7, 8 , 10, 12, 24 (day 2), 36 (day 2), 48 (day 3), 60 (day 3), 72 (day 4) and 84 (day 4) hours after the dose). The determination of the concentration of the active compound is carried out as described in Wilken et al., Diabetologia 43 (51): A143, 2000. The derived pharmacokinetic parameters are calculated from the data of the concentration time for each individual subject by the use of non-compartment methods, using the commercially available software WinNonlin Version 2.1 (Pharsight, Cary, NC, USA). The terminal elimination ratio constant is estimated by logarithmic linear regression in the linear part of the logarithmic terminal of the Concentration time curve, and is used to calculate the elimination of half-life. Analogs and derivatives of stable GLP-1 are described in WO 98/08871 (analogs with lipophilic substituent) and in WO 02/46227 (analogs fused to serum albumin or to Fc portion of an Ig). In another embodiment, the GLP-1 agonist is formulated to have a half-life in a man, as discussed above, of at least 10 hours. This can be obtained by sustained release formulations known in the art. In yet another embodiment the GLP-1 agonist is exendin-4 or exendin-3, an exendin-4 or exendin-3 analogue or a derivative of any of these. Examples of exendins as well as analogs, derivatives, and fragments thereof included within the present invention are those described in WO 97/46584, USA. ,424,286 and WO 01/04156. US 5,424,286 describes a method for stimulating the release of insulin with an exendin polypeptide. The described exendin polypeptides include HGEGTFTSDLSKQMEEEAVRLFIEWLKNGGX [SEQ ID NO. twenty]; where X = P or Y, and HX1X2GTFITSDLSKQMEEEAVRLFIEWLKNGGPSSGAPPPS [SEQ ID DO NOT. twenty-one]; wherein X1X2 = SD (exendin-3) or GE (exendin-4)).

WO 97/46584 describes truncated versions of exendin peptides. The described peptides increase the secretion and biosynthesis of insulin, but reduce those of glucagon. The WO 01/04156 describes analogs and derivatives of exendin-4 as well as the preparation of these molecules. The exemplified exemplified-4 fusion analogs for serum albumin or Fc portion of an Ig se. describe in WO 02/46227. In one embodiment, the exendin-4 analog is HGEGTFTSDLSKQMEEEAVRLFIEWLKNGGPSSGAPPSKKKKKK [SEQ ID? O. 22]. In yet another embodiment the GLP-1 agonist is an analogue / derivative of stable exendin-4. He _. "analogue / derivative of stable exendin-4", as used herein, refers to an exendin-4 (1-39) analogue or an exendin-4 (1-39) analogue derivative which exhibits an mean plasma elimination life in vivo of at least 10 hours in a man, as determined by the method described above by a "stable GLP-1 analogue / derivative". In yet another modality, the GLP-1 agonist is Aib8'35 GLP-1 (7-36) amide (Aib = isobutyric acid a-amino). In still another embodiment, the GLP-1 agonist is Ser38, Lys39'40'41'42'43'44-Exendin-4 (1-39) amide. In yet another embodiment the GLP-1 agonist is selected from the non-peptide small molecule GLP-1 agonists described in WO 00/42026. An amino acid portion of a GLP-1 agonist can be prepared by a variety of methods known in the art such as solid phase synthesis, purification of GLP-1 agonists from natural sources, recombinant technology, or a combination of these methods. See, for example, U.S. Patent Nos. 5,188,666, 5,120,712, 5,523,549, 5,512,549, 5,977,071, 6,191,102, Dugas and Penney 1981, Merrifield, 1962, Stewart and Young 1969, and references cited therein. GLP-1 agonist derivatives can be produced by appropriate derivatization of an appropriate column produced, for example, by recornbinating DNA technology or peptide synthesis (e.g., Merrifield-type solid phase synthesis) using methods known in the art of Peptide synthesis and peptide chemistry. A "gastrin compound" is intended to refer to any compound, including peptides and non-peptide compounds, that are fully or partially associated with and / or activate the gastrin / CCKB receptor. In aspects of the invention, a gastrin compound is selected so as to have an appropriate IC50, for example, an IC50 of about 0.7 nM to a gastrin / CCKB receptor, as measured for methods known in the art (see Singh et al. al (1995) J. Biol. Chem. 270: 8429-8438, and Kopin et al (1995) J. Biol. Chem. 270: 5019-5023 which describe in vitro cell growth assays, and receptor binding assays as described in Singh et al (1995) J. Biol. Chem. 270: 8429-8438, and Kopinetal (1995) J. Biol. Chem. 270: 5019-5023). A gastrin compound can also be selected based on other criteria such as activity, average life etc. as discussed in the present. A "gastrin compound" includes, without limitation, the various forms of gastrin, such as gastrin 71, gastrin 52, gastrin 34 (large gastrin), gastrin 17 (small gastrin), gastrin 14, and gastrin 8 (mini gastrin), pentagastrin, tetragastrin, and fragments, analogs, and derivatives thereof. The sequences for gastrins that include large gastrin-34 (Bonato et al, 1986, Life Science 39: 959) and small gastrin-17 (Bentley et al (1966) Nature 209: 583) are known in the art, and some are shown in SEQ ID NOs. 11 through 18. In particular, the sequences for gastrins include gastrin 71 of SEQ ID NO. 15, gastrin 52 of SEQ ID NO. 16, gastrin 34 (large gastrin) of SEQ ID NO. 11 or 12, gastrin 17 (small gastrin) of SEQ ID NO. 13 or 14, gastrin 14 of SEQ ID NO. 17, and gastrin 6 of SEQ ID? O. 18 or 19. Gastrin-34 is essentially an extension of an amino acid sequence at the terminal end? of gastrina-17. Large gastrin is split in vivo to perform gastrin-17. Glp on the terminal end? of a gastrin is pyroglutamate, which is a naturally cyclized glutamate. In various embodiments, where the cysteine or lysine is added to a term of gastrin having a pyroglutamate, the pyroglutamate is replaced with a glutamate, or the pyroglutamate is removed. A gastrina 34 or gastrin-17 can be used in the invention where there is a methionine or a leucine at position 15, as shown in SEQ ID NOs: 6-9 herein. Examples of gastrin compounds that can be used in the present invention include the compounds described in US Pat. 6,288,301. In some applications of the invention, a gastrin compound may be selected to be a peptide agonist or non-peptide or partial agonist of the gastrin receptor such as A71378 (Lin et al., Am. J. Physiol. 258 (4 Pt 1) : G648, 1990). In some applications of the invention, a gastrin compound can be selected that is a gastrin / CCKB receptor ligand that includes but is not limited to cholecystokinin (CCK), such as CCK 58, CCK 33, CCK 22, CCK 12 and CCK 8, and the like In certain aspects, a gastrin compound may be an active analog, fragment or other modification which, for example, carries the amino acid sequence with an endogenous mammalian gastrin, eg, carries 60% identity of sequence, or 70% identity, or 80% identity Such compounds also include substances that increase the secretion of endogenous gastrins, cholecystinins or similarly active peptides from tissue storage sites, examples of these being the gastric-release peptide, Omeprazole that inhibits acid secretion gastric, and soybean trypsin inhibitor that increases CCK stimulation. The "gastrin compound" includes a modified form of. gastrin, which includes but is not limited to, modified form of gastrin 71 [SEQ ID NO. 15], gastrin 52 [SEQ ID NO. 16], gastrin 34 (large gastrin) [SEQ ID NO. 11 or 12], gastrin 17 (small gastrin) [SEQ ID NO. 13 or 14], gastrin 14 [SEQ ID NO. 17], gastrin 8, gastrin 6 [SEQ ID NO. 18], pentagastrin, and tetragastrin. The modified gastrin preferably comprises TrpMetAspPhe-NH2 [SEQ ID NO. 26] or TrpLeuAspPhe-NH2 [SEQ ID NO. 27]. In aspects of the invention, the modified gastrin comprises at least 1-34, 18-34 or 29-34 amino acids of SEQ ID NO. 11 or 12, or 1-17, 2-17, 12-17, or 14-17 amino acids of SEQ ID NO. 13 or 14. The modified gastrin compounds for use in the present invention comprise the modified gastrin compounds described in PCT / CA03 / 01778, E.U.A Serial No. 10 / 719,450 and patent application E.U.A. No. 60 / 519,933 incorporated in its entirety by reference. In particular, the modified gastrin may be a gastrin derivative or an analog comprising a minimum sequence of 6 amino acids (from the C terminal end) of gastrin, in particular 1 to 34, 18 to 34 or 29-34 residues of amino acids of SEQ ID NO: 11 or 12, or 1-17.2- 17, 12-17, or 14-17 amino acid residues of SEQ ID NO. 13 or 14, and comprises a reactive group capable of undergoing an addition reaction. Examples of reactive groups include, without limitation, thiols, alpha amino groups, epsilon amino groups, carboxyl groups or aromatic rings. The reactive group is generally capable of ligating a gastrin sequence, directly or indirectly by means of a reficulating agent and / or spacer region, to the carrier. The reactive group can be introduced by adding - or 'replacing an amino acid comprising the group. reactive, for example when adding cysteine or lysine. Thus, the modified gastrin may comprise a gastrin sequence (e.g., gastrin-34 or gastrin 17) wherein at least one reactive amino acid (e.g., cysteine or lysine) is added or substituted. The addition of the reactive amino acid may be in a terminal region, in particular an N-terminal region. The modified gastrin may also optionally comprise a spacer. The spacer can interact with the reactive group, for example, an amino acid comprising the reactive group. The spacer can be one or more amino acids, peptides, peptidomimetics, or small organic molecules. The spacer may comprise at least one amino acid, preferably at least two, three, four or five amino acids and in certain embodiments is a sequence of several amino acids, including without limitation alanine or glycine. The spacer may comprise alternative amino acids (eg, glycine and / or alanine), non-alternating amino acids, a random sequence or a particular sequence. By way of example, the spacer can be synthesized as part of, or chemically linked to, an amino acid of the gastrin sequence. The modified gastrin may optionally comprise a crosslinking agent. The crosslinking agent may comprise a homobifunctional or heterobifunctional portion for interaction directly or indirectly with gastrin, spacer and / or reactive group. The crosslinking agent may interact with the gastrin sequence or the spacer, or may be added to the reactive group at the end (in particular N-terminal) of the modified gastrin. The crosslinking agent can be any agent that can bind the gastrin sequence and the carrier directly or via a spacer. Examples of homobifunctional crosslinking agents include without limitation amino group directing homobifunctional crosslinking reagents such as bisimidates (for example, methyl acetimidate-HCl), bifunctional aryl halides (eg, 1,5-dichloro-2,4-dinitrobenzene). ), bifunctional acylating agents (e.g., diisocyanates), bifunctional sulfonyl halides (e.g., phenol-2, - disulfonylchloride), bifunctional acylazides (e.g., tartril diazide), dialdehydes (e.g., glutaraldehyde), and diketones (e.g., 2,5-hexanedione). Examples of heterobifunctional crosslinkers include amino and sulfhydryl group which direct bifunctional reagents (for example, N-succinimidyl-3- (2-pyridyldithiopropionate, carboxyl and any sulfhydryl or amino group that directs bifunctional reagents (e.g., p-diazoacetate) nitrophenyl), and carbonyl and sulfhydryl group driving bifunctional reagents (e.g., 1- (aminooxy) -4- [3-nitro-2-pyridyl) dithio)] butane). The modified gastrin may optionally comprise a carrier which may be a polymer. A carrier can be a polymer or amino acids (proteins), sugars (polysaccharides), nucleosides, synthetic polymers and mixtures thereof. The protein carrier can be a protein found in the circulatory system. Examples of protein carriers found in the circulatory system, in particular in the human circulatory system, include without limitation plasma components such as serum, purified whey proteins such as albumin (in particular human serum albumin), transferrin, or a immunoglobulin, red cell cell proteins such as glycophorin A and AE-1, sugar binding proteins such as lectin, inactivated enzymes, phosphate and sulfate linked proteins, and proteins bound to lipid. Examples of other suitable polymeric carriers include, without limitation, celle and derivatives thereof, starch and derivatives thereof, heparin and derivatives thereof, and synthetic polymers such as polyethylene glycol (PEG) and dextran, and derivatives thereof. The carriers can be linked to the gastrin or spacer as reactive groups on, or introduced into, the carrier, gastrin, and / or spacer. For example, carriers can be covalently linked to reactive groups (such as thiol groups, alpha and epsilon amino groups, carboxyl groups or aromatic groups) in the gastrin or spacer that can be presented or added by chemical modification of the gastrin or spacer. In certain aspects of the invention, the modified gastrin may comprise the gastrin of SEQ ID NOS 11, 12, 13, 14, 17, or 18 and a carrier. The group of modified gastrin compounds include compounds having an amino acid sequence comprising from the amino terminus Z-Ym-Xn-AA? -AA2-AA3-AA-AA5-AA6, wherein AAX is Tyr or Phe, AA2 is Gly, Ala, or Ser, AA3 is Trp, Val, or lie, AA4 is Met or Leu, AA5 is Asp or Glu, and AA6 is Phe or Tyr and wherein AA6 is optionally amidated; Z is a carrier, in particular a polymer and when the polymer is a protein, Z is an amino acid sequence; Ym is an optional spacer region comprising m 'residues of amino acids of the small neutral amino acid, which includes, but is not limited to serine and alanine, and X is any consecutive portion of residues 1-28 (= n) of SEQ ID NO: 11 or 12 or 1-11 of SEQ ID. DO NOT. 13 or 14, with the proviso that the gastrin compound binds the gastrin / CCKB receptor. Generally, m is 0 to about 20 residues. In one aspect, Z is a protein, in particular a protein of the circulatory system, more particularly a whey protein, still more particularly albumin, even more particularly human serum albumin. In embodiments, X is one or more amino acid residues from position 18 to position 20 of SEQ ID NO: 11. Therefore, the gastrin compounds by virtue of the presence of X, may have any of the sequences of gastrina from positions 18-28, 19-28, 20-28, 21-28, etc. The gastrin compound optionally contains an amino acid spacer (Y) of length m, and m is 0 to about 20 residues. In embodiments, X is one or more amino acid residues from position 1 to 11 or 2 to 11 of SEQ ID NO: 13 or 14-. Therefore, the gastrin compounds by virtue of the presence of X, can have any of the gastrin sequences from positions 2 to 11, 3 to 11, 4 to 11, 5 to 11, etc. The gastrin compound optionally contains an amino acid spacer (Y) of length m, and m is O to about 20 residue. A gastrin compound includes a modified gastrin compound of the formula Xn-AA? -AA2-AA3-AA4-AA5 ~ AA6 where there is no spacer (Y) and m is 0, which may also comprise a bifunctional crosslinking agent for the interaction or linked to a Z carrier, wherein Z 'further comprises a non-proteinaceous polymer such as dextran or PEG. A modified gastrin compound particularly described herein may further comprise an amino terminal cysteine or lysine residue. In some embodiments of the modified gastrin compounds described herein, the gastrin component contains at least one amino acid residue 29-34 of SEQ ID NO: 11 or 12, and this is associated with a polymer, a lipid or a carbohydrate. The polymer can be a synthetic polymer or it occurs naturally. The term "polymer" includes an amino acid polymer protein, and is not limited to a synthetic polymer. The polymer can be a polyethylene glycol (PEG) or a dextran. A modified gastrin compound may be based on SBQ ID NO: 11 or 12 or "large" gastrin-34 and has a residue at position 32 which is a methionine or a leucine, respectively. Another preferred modified gastrin compound comprises a structure C-Ym-X, where C is Cys or Lys, Ym is an optional spatial region comprising residues of amino acids m of a small neutral amino acid; and X is at least six amino acid residues comprising at least positions 12-17 of gastrin-17 (SEQ ID NO: 13 or 14) or at least positions 29-34 of gastrin-34 (SEQ ID NO: 11 or 12). This modified gastrin compound may further comprise a bifunctional crosslinking agent wherein one reactive portion of the crosslinking agent is covalently linked to C, and the other reactive portion is covalently linked to a polymer or protein. In a particular aspect of the invention AA? -AA2-AA3-AA4-AA5-AA6 in a modified gastrin compound is Tyr-Gly-Trp-Met-Asp-Phe [SEQ ID NO. 23] or Tyr-Gly-Trp-Leu-Asp-Phe [SEQ ID NO. 24]. The gastrin compounds can be synthesized by chemical synthesis using techniques known in the chemistry of proteins such as solid phase synthesis (Merrifield, 1964, J. Am. Chem. Assoc. 85: 2149-2154) or homogeneous solution synthesis (Houbenweyl, 1987, Methods of Organic Chemistry, ed. E. Wansch, Vol. 15 I and II, Thieme, Stuttgart). The synthesis can be carried out using manual procedures or by automation. Automated synthesis can be carried out, for example, using an Applied Biosystems 431A peptide synthesizer (Perkin Elmer). The gastrin compounds can also be obtained from commercial sources. For example, human gastrin synthetics 17 with methionine or leucine at position 15 are available from Bachem AG, Bubendorf, (Switzerland), and from Research Plus hie (New Jersey, United States of America). A "gastrin / CCK receptor" refers to a number of G protein coupling receptor families that exhibit an affinity-binding characteristic for a cholecystokinin (CCK) including without limitation CCK-8, CCK-8, CCK-33 , Desulfated CCK-4, or gastrins that include no desulfated limitation or sulfated gastrin-17, or pentagastrin, or other CCK or gastrin analog or family members. Examples of gastrin / CCK receptor proteins are CCKA and gastrin / CCKB receptors, in particular a gastrin / CCK receptor. The conditions and / or diseases refer to one or more pathological symptoms or syndromes for which either alone or both, a GLP-1 agonist or a gastrin compound provides a beneficial or therapeutic effect. The condition and / or disease may require the reduction of blood glucose levels, inhibition of gastric acid secretion, inhibition of β-cell apoptosis, stimulation of proliferation or differentiation of β-cells, and reduction of weight in the body. Examples of conditions and / or diseases include but are not limited to dyslipidemia, hyperglycemia, severe hypoglycemic episodes, stroke, left ventricular hypertrophy, arrhythmia, bacteremia, septicemia, irritable bowel syndrome, respiratory distress syndrome, functional dyspepsia, diabetes, catabolic changes after surgery, hyperglycemia induced by tension, gastric ulcers, myocardial infarction, impaired glucose tolerance, hypertension, chronic heart failure, fluid retention, metabolic syndrome and related diseases and disorders, obesity, diabetic complications as well as symptoms of other diseases in which tissue is damaged due to elevated glucose levels including Alzheimer's disease, Parkinson's disease and other degenerative diseases of tissue, related to age, as well as the arteriogenic effects of elevated leptin, for example in patient with impaired glucose tolerance and non-diabetic obese patient. The term "diabetes" as used herein means any manifested symptom of diabetes in any mammal including experimental animal models, and including human forms such as type I and type II diabetes, early stage diabetes and a pre-diabetic condition characterized by gently lowering insulin or gently elevating blood glucose levels. A "pre-diabetic condition" describes a subject who demonstrates a symptom in terms of insulin or glucose levels and / or demonstrates a susceptibility to diabetes or a condition related to family history, genetic predisposition, or obesity in the case of type II diabetes, and includes a subject who previously has diabetes or a related condition and is at risk of it occurring. "Insulinotropic activity" refers to an ability of a substance to stimulate insulin secretion in response to elevated glucose levels to produce or increase the cleavage of glucose by cells and decrease the levels of glucose in the serum and glucose in the blood. blood. The methods known in the art can be used to test insulinotropic activity. For example, the in vitro and in vivo methods can be used to measure the activity bound to the GLP-1 receptor or gastrin receptor-linked activity, receptor activation (See methods described in BP 619,322 for Gelfand et al and US patent No 5,120,712), and / or insulin or C-peptide levels. The compounds, compositions or conjugates described herein have insulinotropic activity if the islet cells secrete insulin in the presence of the compounds, compositions or conjugates above at pooled levels or levels in the absence of - compounds, compositions or conjugates. A compound can be administered to an animal and the insulin concentration It can be monitored for a while. "Islet neogenesis" means the formation of new beta cells by differentiation, which may or may not have characteristics of stem cells which have the ability to reproduce in an unlimited manner.

Detailed Description of the Invention The invention relates to compositions, conjugates and methods that utilize a GLP-1 agonist and a gastrin compound to provide beneficial effects. In particular, the invention relates to compositions, conjugates and methods for the prevention, intervention and / or treatment of a condition and / or disease discussed herein comprising a GLP-1 agonist and a gastrin compound. In aspects of the invention, the compositions, conjugates and methods of the invention provide high beneficial effects, in particular sustained beneficial effects relative to a GLP-1 agonist and / or a gastrin alone compounds. The beneficial effects can be additives or synergistic effects. In aspects of the invention, where the condition and / or disease is diabetes, beneficial effects in particular sustained beneficial effects of a composition, combination treatment, or conjugate of the invention may manifest as one or more of the following: a) An increase in pancreatic insulin levels relative to the levels measured in the absence of the active compounds or for each compound only after administration to a subject with symptoms of diabetes. Preferably, the compounds together include at least about 0.05%, 0.1%, 0.5%, 1%, 2%, 5%, 10%, 15%, 20%, 30%, 33%, 35%, 40%, 45 %, or 50% increase in pancreatic insulin levels in a subject. b) A reduction of an absence of symptoms of inflammation of "islets after administration to a subject with symptoms of diabetes c) A decrease in blood glucose levels relative to the levels measured in the absence of the compounds or for each compound only in subjects with symptoms of diabetes Preferably, the compounds include at least about 1%, 2%, 5%, 10%, 15%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, or 90% decrease in blood glucose levels More preferably, the compounds produce blood glucose levels around or close to common levels in a normal subject d) An increase in C-peptide levels relative to the levels measured in the absence of the compounds or for each compound only in subjects with symptoms of diabetes Preferably, the compounds together induce at least around 0.05%, 0.1%, 0.5%, 1%, 2%, 5%, 10%, 15%, 20%, 30%, 33%, 35%, 40%, 45%, or 50% increase in C-peptide levels e) Maintenance of blood glucose levels around normal for a prolonged period of time. f) The maintenance of hemoglobin Ale or glycated hemoglobin around normal levels for a prolonged period of time, in particular maintaining a% hemoglobin Ale a between 6-8%, more particularly at around 7%. g) A reduction in destruction of beta cells. Preferably the compounds induce at least about 1%, 2%, 5%, 10%, 15%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, or 90% reduction. h) An increase in the function of beta cells.

Preferably the compounds induce at least about 1%, 2%, 5%, 10%, 15%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, or 90% increase in beta cell function. i) A reduction, prevention or loss of the relationship of the progression of the disease in a subject with diabetes. j) A reduction or prevention of the development of severe hyperglyceria and ketoacidosis with symptoms of diabetes. k) An increase in survival in a subject with symptoms of diabetes. In embodiments of the invention, the beneficial effects or sustained beneficial effects comprise or consist essentially of two, three, four, five, six, seven, eight, nine, ten or eleven, from a to k). , beneficial effects or sustained beneficial effects comprise or consist essentially of a), b), and c), a), b), c), and d), a), b), c), d), and e); ), b), c), d), e), and f), a), b), c), d), e), f), and g), a), b), c), d), e), f), g), and h), a), b), c), d), e), f), g), h), yi), a), b), c), d) , e), f), g), h), i) and j), a), d), and e), a), d), e), and h), a), d), e), h) , yi), a), d), e), h), i), and j), a), b), c), d), e), h), i), and j) a), b ), c), d), e), h), i), j), and k), b), c), d), y), b), c), d), e), h), i), and j), and, b), h), i) and j) One or more of these beneficial effects or sustained beneficial effects can be demonstrated in a diabetic subject or disease model, p or example, a non-obese mouse (NOD) with symptoms of diabetes, using standard methods known to those skilled in the art. For example, commercially available methods and kits can be used to test pancreatic insulin levels, glucose levels, levels in peptide C and hemoglobin Ale A gastrin compounds can be selected by particular embodiments in the present invention and for provide a specific beneficial effect based on characteristics including this insulinotropic activity, the ability to increase the activity of a GLP-1 agonist (in particular to elevate the insulinotropic effects of a GLP-1 agonist), and / or increase the physical or chemical stability of a GLP-1 agonist. A gastrin compound can also be selected based on this ability to stimulate beta cell proliferation / differentiation and half-life in vivo. In one aspect of the invention, a gastrin compound that is used in the methods, compositions and conjugates of the invention is grastin 17 and analogs and derivatives thereof, associated with a polymer. In a particular aspect, the gastrin compound is synthetic human gastrin I having 17 amino acid residues with a Leu residue in an amino acid at position 15 [SEQ ID NO. 14]. In another aspect of the invention, a gastrin compound that is used in the methods, compositions and conjugates of the invention is gastrin 34 and analogs and derivatives thereof. In a particular aspect, the gastrin compound is a synthetic human gastrin 34 with methionine or leucine at position 32 [SEQ ID NO. 11 or 12]. In a further aspect of the invention, a gastrin compound that is used in the methods, compositions and conjugates of the invention is gastrin 34 or gastrin 17 or portions thereof, directly or indirectly interacting with or associated with a whey protein, in particular albumin or an immunoglobulin. More particularly human serum albumin. In particular aspects of the invention, a gastrin compound comprises a synthetic human gastrin 34 having 2-34 amino acid residues of SEQ ID NO. 11 or 12, and optionally a cysteine at the N-terminal and / or a carrier; the synthetic human gastrin has 1-17 amino acid residues with a Leu residue at the amino acid position 15 [SEQ ID NO. 14] -and optionally a cysteine residue at the N-terminus; and a synthetic human gastrin having amino acid residues 2 through 17 or 5-17 of SEQ ID NO. 13 or 14, optionally with a N-terminal cysteine residue and / or a carrier (eg, PEG or human serum albumin) bound via a spacer [eg, Gly-Ala-Gly-Ala-Gly-Ala- Gly-Ala-Gly-Ala, that is, (GA) 5] [SEQ ID NO. 25], in particular, a synthetic human gastrin having amino acid residues 2 through 17 or 5-17 of SEQ ID NO. 13 or 14, with a polymer of human serum albumin (HSA) bound by means of a spacer Gly-Ala-Gly-Ala-Gly-Ala-Gly-Ala-Gly-Ala [that is, (GA) 5], and optionally a C-terminal residue in the N-terminus. A GLP-1 agonist can be selected for particular applications in the present invention based on one or more of the following characteristics: ability to bind to the GLP-1 receptor, preferably with a constant affinity Kd of less than about 1 μM, more preferably at least about 100 μM; ability to initiate a signal transduction path resulting in insulinotropic activity; insulinotropic activity; stimulation of beta cell proliferation / differentiation; resistance for DPIV cleavage, and an in vivo half-life of at least about 15 minutes to 24 hours, preferably 2 to 10 hours or 2 to 8 hours in humans using conventional methods (see for example, the method described in the patent of USA 2003/0144206). In aspects of the invention, the GLP-1 agonist is a naturally truncated GLP-1 polypeptide (GLP-1 (7-36) or ((GLP-1 (7-37)), or an analogue or derivative thereof. that these truncated GLP-1 agonists occur naturally are represented in SEQ ID NOs 4, 5, and 6. In certain aspects of the invention, a GLP-1 agonist can have the amino acid sequence of SEQ ID NOs. , 2, or 3 modified so that the amino acid residues at positions 1-20, preferably 1-15, more preferably 1-10, more preferably 1-5 differ from the sequences of SEQ ID NOs. 3. In one embodiment of the invention, the GLP-1 agonist is an analog of GLP-1 (7-37) or GLP-1 (7-36) which has the minus 10 amino acid residues that are different from those in GLP-I (7-37) or GLP-I (7-36), less than 5 amino acid residues that are different from those in GLP-I (7-37) or GLP-I (7-36), less than 3 amino acid residues that are different from those in GLP-1 (7-37) or GLP-I (7-36), preferably only one amino acid residue that is different from the sequence of GLP-1 (7-37) or GLP-1 (7-36). The GLP-1 agonist which may have specific utility in the present invention includes polypeptides where one or more amino acids have been added to the N-terminal and / or C-terminus of GLP-I (7-37) or GLP-I (7-36). ). Preferably, about one to six amino acids can be added to the N-terminus and / or about one to eight amino acids can be added to the C-terminus. In certain applications the GLP-1 agonists are selected to have up to 39 amino acids. The amino acids at positions 1-6 of an extended GLP-1 agonist can be selected to be therein or amino acid substitutions are retained at the corresponding positions of GLP-I (7-37) or GLP-I (7- 36) precursor. The amino acids at positions 38-45 of an extended GLP-1 agonist may be selected to be the same or amino acid substitutions are retained at the corresponding positions of exendin-3 or exendin-4 (SEQ ID No. 7 and 8 respectively ). In aspect of the invention a GLP-1 agonist is used to understand an analogue at position 8 wherein the column for such analogues or fragments thereof contains an amino acid other than alanine. The amino acid at position 8 can be selected from "glycine, valine, leucine, isoleucine, serine, threonine or methionine In one embodiment a GLP-1 agonist is an insulinotropic analog of GLP-I (1-37), for example, Met 8 -GLP-1 (7-37), where the alanine at position 8 is replaced by methionine and the amino acid residues at position 1 to 6 have been deleted and Arg34-GLP-1 (7-37) where the Valine at position 34 is replaced with arginine and the amino acid residues at position 1 through 6 have to be deleted.

In another embodiment, GLP-1 agonists are selected from those having the amide sequences GLP-1 (7-37) OH and GLP-1 (7-36), and analogs at the corresponding 8-position where the column for such analogs it contains an amino acid other than alanine. The amino acid at position 8 can be selected from glycine, valine, leucine, isoleucine, serine, threonine or methionine, preferably valine or glycine. Analogs can additionally contain (a) an amino acid at position 22 selected from glutamic acid, lysine, aspartic acid, arginine, and preferably glutamic acid or lysine; (b) an amino acid at position 30 is selected from glutamic acid, aspartic acid, serine or histidine; (c) an amino acid in position 37 is selected from lysine, arginine, threonine, glutamic acid, aspartic acid, serine, tryptophan, tyrosine, phenylalanine, or histidine; and / or (d) amino acid at position 27 is selected from alanine, lysine, arginine, tryptophan, tyrosine, phenylalanine, or histidine. A group of GLP-1 analogs and derivatives for use in the present invention comprises the GLP-1 agonist described in the US patent. No. 5,545,618 and U.S. Patent Application. Serial No. 20040018975. Analogs include activated GLP-1 peptides, 7-34, 7-35, 7-36, and 7-37 that have amino acid substitutions at positions 7-10 and / or are terminal C truncations. and / or contain various other amino acid substitutions in the basic peptide. Preferred analogs include those with amino acid substitutions D at positions 7 and 8 and / or N-alkylated or N-acylated amino acids at position 7 from those that are particularly resistant to in vivo degradation. In aspects of the invention, a GLP-1 agonist comprises a peptide comprising or selected from the group consisting of GLP-1 (1-38); GLP-1 (1-39), GLP-1 (1-40), GLP-1 (1-41), GLP-1 (7-38), GLP-1 (7-39), GLP-1 (7 -40), 'and GLP-1 (7-41). In another aspect of the invention at least one amino acid of a GLP-1 agonist having at least one substituent linked directly or indirectly (for example, by middle of a spacer such as? -Glu or ß-Ala). A substituent is generally selected to make the action profile of the longer precursor agonist GLP-1, make the agonist GLP-1 more metabolically and physically establish and / or increase the solubility of the GLP-1 agonist. An example of a particular substituent is an amide, a carbohydrate, and a lipophilic substituent. A lipophilic substituent includes but is not limited to an alkyl group, a group which has a carboxylic acid group, an acyl group of a straight or branched chain, fatty acid or alkane such as tetradecanoyl, hexadecanoyl. The particular compositions, conjugates and treatments of the invention using GLP-1 agonists with lipophilic substituents such as those described in WO 99/43341 (Novo Nordisk) and US 2003 / 0119734A1 (Novo Nordisk). In particular aspects of the invention an agonist "GLP-1 is a GLP-1 (7-36) -amide or Tyr31-exendin-4 (1-31) -amide.

Certain aspects of the invention produce an agonist GLP-1 which is a derivative of GLP-1 (7-36) or GLP-1 (7-37) which comprises a lipophilic substituent. In one embodiment, the GLP-1 agonist is Arg34Lys2S (Ne (? -Glu (Na-hexadecanoyl))) -GLP-1 (7-37). In embodiments of the invention, the GLP-1 agonist comprises or is selected from the group consisting of: Gly8-GLP-1 (7-36) -amide, Gly8-GLP-1 (7-37), Val8-GLP-1 (7-36) -amide, Val8-GLP-I (7-37), Val8Asp22-GLP-I (7-36) -amide, Val 8 Asp22-GL-1 (7-37), Val8Glu22-GLP-I (7-36) -amide, Val8Glu22-GLP-I (7-37), Val8Lys22-GLP-I (7-36) -amide, Val8Lys22-GLP-I (7-37) -Val8Arg22-GLP-1 (7-36) -amide, Val 8Arg22 -GLP-I (7-37), Val8His22-GLP-I (7-36) -amide, -Val8His22-GLP-I (7-37), Arg26-GLP-1 (7-37); Arg34- "GLP-1 (7-37); Lys36-GLP-1 (7-37); Arg26.3 Lys3S-GLP-I (7-37); Arg26 '34 -GLP -1 (7-37); 'Arg26'34Lys40-GLP-I (7-37); Arg26Lys36-GLP-1 (7-37); Arg34Lys36-GLP-I (7-37); Val8Arg22-GLP-I (7-37); Met8Arg22-GLP-I (7-37); Gly8His22-GLP-I (7-37); Val8His22-GLP ~ l (7-37); Met8His22-GLP-I (7-37); His37-GLP-1 (7-37); Gly8-GLP-1 (7-37); Val8-GLP-1 (7-37); Met8-GLP-1 (7-37); Gly8Asp22-GLP-I (7-37); Val 8Asp22-GLP-I (7-37); Met8Asp22-GLP-I (7-37); Gly8Glu22-GLP-I (7-37); Val8-GLP-1 (7-37), - Gly8Lys22-GLP-I (7-37); Val8Lys22-GLP-1 (7-37); Glu22Met8Glu22-GLP-I (7-37); Gly8Lys22-GLP-I (7-37); Val 8Lys22-GLP-1 (7-37); Met6Lys22-GLP-I (7-37); Gly8Arg22-GLP-I (7-37); Val8Lys22His37-GLP-I (7-37); Gly8Glu22His37-GLP-I (7-37) Val8Glu22His37- GLP-1 (7-37); Met8Glu22His37-GLP-I (7-37) Gly8Lys22His37-GLP-I (7-37); Met8Lys22His37-GLP-l (7-3.7) Gly8Arg22His37-GLP-I (7-37); Val8Arg22His37-GLP-I (7-37) Met8Arg22His37-GLP-I (7-37); Gly8His22His37-GLP-l (7-37) Val8His22His37-GLP-I (7-37); Met8His22His37-GLP-I (7-37) Gly8His37-GLP-I (7-37); Val8His37-GLP-I (7-37); Met8His37-GLP-I (7-37); Gly8Asp22His37-GLP-I (7-37); Val8Asp 2His37-GLP-I (7-37); Met8Asp22His37-GLP-I (7-37); Arg26-GLP- (7-36) -amide; Arg34-GLP-1 (7-36) -amide; Lys36-GLP-1 (7-36) -amide; Arg26'34Lys36-GLP-l- (7- 36) -amide; Arg26 '34 -GLP -1 (7-36) -amide; Arg26'3 Lys40-GLP-I (7-36) -amide; Arg26Lys36-GLP-I (7-36) -amide; Arg34Lys36-GLP-I (7-36) -amide; Gly8-GLP-1 (7-36) -amide; Val8-GLP-1 (7-36) -amide, - Met8-GLP-1 (7-36) -amide; Gly8Asp22-GLP-I (7-36) -amide; Gly8Glu22His37- GLP-1- (7-36) -amide; Val 8Asp 2 -GLP-1 (7-36) -amide; Met8Asp2-GLP-1 (7-36) -amide; Gly8Glu22-GLP-I (7-36) -amide; Val8Glu2-GLP-1 (7-36) -amide; Met8Glu22-GLP-I (7-36) -amide; Gly8Lys22-GLP-I (7-36) -amide; Val8Lys22-GLP-I (7-36) -amide; Met8Lys 2-GLP-I (7-36) -amide; Gly8His22His37-GLP-I (7-36) -amide; Gly8Arg2-GLP-l (7-36) -amide; Val8Arg 2-GLP-1 (7-36) -amide; Met8Arg2-GLP-1 (7-36) -amide; Met8Arg22His37-GLP-1 (7-36) -amide; Gly8His22-GLP-I (7-36) -amide; Val8His22-GLP-I (7-36) -amide; Met8His22-GLP-I (7-36-amide His37-GLP-1 (7-36) -amide; Val8Arg22His37-GLP-I (7-36) -amide Met8Arg22His37-GLP-1 (7-36) -amide; Gly8His37-GLP-l (7-36) -amide Val 8His37-GLP-I (7-36) -amide; Met8His37-GLP-l (7-36) -amide Gly8Asp22His37-GLP-I (7-36) -amide; Val8Asp22His37-GLP-I (7-36) -amide; Met8Asp22His37-GLP-I (7-36) -amide; Val8Glu22His37-GLP-I (7-36) -amide; Met8Glu22His37-GLP-I (7-36) -amide; Gly8Lys22His37-GLP-1 (7-36) -amide; Val8Lys22His37-GLP-I (7-36) -amide; Met8Lys22His37-GLP-1 (7-36) -amide; Gly8Arg22His37-GLP-I (7-36) -amide; Val8His22His37-GLP-I (7-36) -amide; Met8His22His37-GLP-I (7-36) -amide; Val8-GLP-1 (7-37) OH, Gly8-GLP-1 (7-37) OH, Glu22-GLP-1 (7-37) OH, Arg22-GLP-I (7-37) OH, Arg22- GLP-1 (7-37) OH, Lys22-GLP-1 (7-37) OH, Cys22-GLP-1 (7-37) OH, Val8-Glu22-GLP-1 (7-37) OH, Val8- Asp22-GLP-1 (7-37) OH, Val8-Arg22-GLP-1 (7-37) OH, Val8Lys 2-GLP- 1 (7-37) OH, Val8-Cys22-GLP-I (7-37) OH, Gly8-Glu22-GLP-1 (7-37) OH, Gly8-Asp 2-GLP-I (7-37) OH , Gly8 -Arg22- GLP -1 (7-37) OH, Gly8-Lys22- GLP-I (7-37) OH, Gly8-Cya22-GLP-1 (7-37) OH, Glu2-GLP-1 (7 -36), NH2, ASP22-GLP-1 (7-36) NH2, Arg22-GLP-1 (7-36)? H2, Lys22-GLP-1 (7-36)? H2, Cys22-GLP-1 (7-36) NH2, Val 8 -Glu22-GLP -1 (7-36)? H2, Val8-Asp22-GLP-1 (7-36)? H2, Val 8 -Arg22- GLP-1 (7-36)? H2, Val8 -Lys22-GLP-1 (7-36)? H2, Val8-Cys22-GLP-1 (7-36)? H2, Gly8-Glu22-GLP-1 (7-36)? H2, Gly8Asp22-GLP-I ( 7-36)? H2, Gly8 -Arg22- GLP -1 (7-36)? H2, Gly8-Lys 2-GLP-1 (7-36)? H2, Gly8 -Cys22- GLP-1 (7-36)? H2, Lys23-GLP-1 (7-37) OH, Val8-Lys23-GLP-I (7-37) OH, Gly8-Lys23-GLP-1 (1-31) OH, His24-GLP-1 (7-37) OH, Val8-His24-GLP-1 (7-37) OH, Gly8-His24-GLP-1 (7-37) OH, Lys24-GLP-I (7-37) OH, Val8-Lys2-GLP-1 (7-37) OH, Gly8-Lys23-GLP-1 (7 -37) OH, Glu30-GLP-1 (7-37) OH, Val8-Glu30-GLP-1 (7-37) OH, Gly8-Glu30-GLP-1 (7-37) OH, Asp30-GLP-1 (7-37) OH, Val8-Asp30-GLP-1 (7-37) OH, Gly8-Asp30-GLP-1 (7-37) OH, Gln30-GLP-1 (7-37) OH, Val8-Gln30 -GLP-1 (7-37) OH, Gly8-Gln30GLP-I (7-37) OH, Tyr30-GLP-I (7-37) OH, Val8-Tyr30-GLP-I (7-37) OH, Gly8 -Tyr30- GLP -1 (7-37) OH, Ser30-GLP-1 (7-37) OH, Val8-Ser30-GLP-1 (7-37) OH, Gly8-Ser30-GLP-1 (7-37) ) OH, His30-GLP-1 (7-37) OH, Val8His30-GLP-I (7-37) OH, Gly8-His30-GLP-1 (7-37) OH, Glu34-GLP-1 (7-37) ) OH, Val8Glu34-GLP-I (7-37) OH, Gly8-Glu34-GLP-1 (7-37) OH, Ala34-GLP-1 (7-37) OH, Val8-Ala34-GLP-I (7 -37) OH, Gly8 -Al a34- GLP-1 (7-37) OH, Gly3-GLP-1 (7-37) OH, Val8-Gly34-GLP-l (7-37) OH, Gly8-Gly34- OLP-l (7-37) OH, Ala35-GLP-l (7-37) OH, Val8-Ala35-GLP-1 (7-37) OH, Gly 8- Al a35- GLP -1 (7-37) OH, Lys35-GLP-1 (7-37) OH, Val8-Lys35-GLP-1 (7-37) OH, Gly8- Lys35-GLP-1 (7-37) OH, His35-GLP-1 (7-37) OHVal8-His35-GLP-I (7-37) OH, Gly8-His35-GLP-I (7-37) OH, Pro35-GLP-I (7-37) OH, Val8-Pro35-GLP-1 (7-37) OH, Gly8-Pro35-GLP-I (7-37) OH, Glu35-GLP-1 (7-37) OH, Val8-Glu35-GLP ~ l (7-37) OH, Gly8-Glu35-GLP-1 (7-37) OH, Val8-Ala27-GLP-1 (7-37) OH, Val8-His37-GLP- 1 (7-37) OH, Val8-Glu22-Lys23-GLP-1 (7-37) OH, Val8-Glu22-Glu23-GLP-1 (7-37) OH, Val8-Glu22-Ala27-GLP-1 ( 7-37) 0H, Val8-Gly3-Lys3s-GLP-1 (7-37) OH, Val8-His37-GLP-I (7-37) OH, and Gly8-His37-GLP-1 (7-37) OH , and derivatives thereof. In a particular embodiment a GLP-1 agonist comprises or is selected from the group consisting of Val8-GLP-1 (7-37) OH, Gly8-GLP-1 (7-37) OH, Glu22-GLP-1 (7- 37) OH, Lys22-GLP-1 (7-37) OH, Val 8 -Glu22-GLP-1 (7-37) OH, Val 8 -Lys22-GLP-1 (7-37) OH, Gly8-Glu22- GLP-1 (7-37) OH, Gly8-Lys22-GLP-1 (7-37) OH, Glu22-GLP-1 (7-36) NH2, Lys22-GLP-1 (7-36) NH2, Val 8 - Glu22- GLP -1 (7-36) NH2 / Val8-Lys22-GLP-1 (7-36) NH2, Gly8-Glu22-GLP-1 (7-36)? H2, Gly8-Lys22-GLP-1 ( 7-36) NH2, Val8-His37-GLP-I (7-37) OH, Gly8-His37-GLP-1 (7-37) OH, Arg3-GLP-1 (7-36)? H2, and Arg34-GLP-1 (7-37) OH. In another particular embodiment, the GLP-1 agonist comprises or is selected from the group consisting of Gly8-GLP-1 (7-37), Val8GLP-l (7-37), Val8Asp22GLP-l (7-37), Val8Glu22GLP -1 (7-37), Val8Lys22GLP-I (7-37), and Val8His22GLP-I (7-37), and analogues and derivatives thereof. In a further particular embodiment, the GLP-1 agonist comprises or is selected from the group consisting of Gly8- GLP1 (7-36) amide, Val8 GLP1 (7-36) amide, Val8Asp22GLPl (7-36) amide, Val8Glu22GLPl (7-36) amide, Val8Lys22GLPl (7-36) amide, and Val8His22GLPl (7-36) amide, and analogs and derivatives thereof. In still further embodiments of the invention, the GLP-1 agonist (e.g., exedin 3 and exedin 4) or an analog, derivative, or fragment thereof. Examples of exendins that can be used in the present invention include without limitation those described in WO 9746584, 0 E.U.A. No. 5,424,286 and WO 01/04156. The Patent E.U.A. No. 5,424,286 describes the use of an exedin polypeptide to stimulate the release of insulin. The exendin polypeptides described in the U.S. Pat. include HGEGTFTSDLSKQMEEEAVRLFIEWLKNGGX where X = P or Y, and 5 HX1X2GTFITSDLSKQMEEEAVRLFIEWLKNGGPSSGAPPPS; where X1X2 = SD (exendin-3) or GE (exendin-4). WO 9746584 describes exendin truncated peptides that increase insulin secretion and biosynthesis, but reduce glucagons. The WO 01/04156 describes analogues and derivatives of exendin-4 and its preparation. In another embodiment, the GLP-1 agonist is an insulinotropic analogue of exendin-4 (1-39), in particular Ser2Asp3-exendin-4 (1-39) wherein the amino acid residues in position 2 and 3 have been replaced. with 5 serine and aspartic acid, respectively (this analogue particular is also known in the art as exendin-3, SEQ ID NO: 7). In certain aspects of the invention, the GLP-1 agonist is a stable GLPXL agonist, in particular a GLP-1 analog or derivative, or a stable exendin-4 or exendin-3 analog or derivative. The pharmaceutical compositions of the invention can be selected so as to provide beneficial effects, in particular statistically beneficial beneficial effects or "sustained" beneficial effects, as compared to a GLP-1 agonist or a gastrin compound alone. Diabetic can be evidenced by one or more of the beneficial effects described herein, in particular one, two, three, four, five, six, seven, eight, nine or ten of the beneficial effects described in a) to j) above. The pharmaceutical composition with beneficial effects, particularly particular statistically significant beneficial effects or sustained beneficial effects, is provided comprising a GLP-1 agonist selected from the group consisting of Gly8-GLP-1 (7-37), Val8GLP-I (7 -37), Val8Asp22GLP-1 (7-37), Val8Glu22GLP-I (7-37), Val8Lys22GLP-I (7-37), Val8His22GLP-I (7-37), Arg34Lys26 (? E (? -Glu (? a-hexadecanoyl))) -GL P-K7-37), Gly8-GLP-1 (7-36) amide, Val8GLP-I (7-36), Val8Asp22GLP-I (7-36) amide, Val8Glu22GLP-I (7-36) amide, Val8Lys22GLP-1 (7-36) amide, and Val8His22GLP-1 (7-36) amide, and a gastrin compound (eg, SEQ ID NO, 11, 12, 13 or 14), optionally associated with a whey protein . In one aspect, the pharmaceutical composition with statistically significant beneficial effects or sustained beneficial effects is provided comprising GLP-1 (7-36) [SEQ ID NO: 5) and gastrin-17 (leu) [SEQ ID NO: 14]. In one aspect, the pharmaceutical composition with statistically significant beneficial effects or sustained beneficial effects is provided comprising Arg34Lys26 (Ne (? -Glu (N01-hexadecanoyl) -GLP-K7-37), and gastrinal7 (leu) [SEQ ID NO: 14], In another aspect, the pharmaceutical composition with beneficially statistically significant effects or sustained beneficial effects is provided comprising Aib8,35GLP-l (7-36) amide or Ser38, Lys39'40'41 '2'3'4 -Exendin -4 (1-39) amide and gastrin-17 (leu) [SEQ ID NO: 14] The pharmaceutical composition with beneficial effects, in particular statistically significant beneficial effects or sustained beneficial effects, is provided comprising a GLP-1 agonist selected from the group consisting of Gly8-GLP-K7-37), Val8GLP-I (7-37), Val8Asp22GLP-I (7-37), Val8Glu22GLP-I (7-37), Val8Lys22GLP-I (7-37) , Val8His22GLP-I (7-37), Arg34Lys26 (Ne (? -Glu (Na-hexadecanoyl))) -GLP-1 (7-37), Gly8-GLP-1 (7-36) amide, Val8GLP-I ( 7-36) amid a, Val8Asp22GLP-l (7-36) amide, Val8Glu22GLP-l (7-36) amide, Val8Lys22GLP-l (7-36) amide, and Val8His22GLP-1 (7-36) amide, and a gastrin compound having an amino acid sequence comprising, from the amino terminal, Z-Ym-Ym-AA? -AA2-AA3-AA-AAs-AAs, in where AAX is Tyr or Phe, AA2 is Gly, Ala, or Ser, AA3 is Trp, Val, or lie, AA4 is Met or Leu, AA5 is Asp or Glu, and AA6 is Phe or Tyr; Z is a polymer and when the polymer is a protein Z is an amino acid sequence; Ym is an optional spacer region comprising amino acid residues m of the small neutral amino acid including but not limited to serine and alanine, and X is any consecutive portion of residues 1-28 of SEQ ID NO: 11 - or 12, or residues 1-11 of SEQ ID NO: 13 or 14, preferably AA-AA2-AA3-AA4-AA5-AA6 is Tyr-Gly-Trp-Met-Asp-Phe or Tyr-Gly-Trp-Leu-Asp-Phe. In a particular embodiment, Z is a whey protein, in particular human serum albumin. In certain aspects of the invention, pharmaceutically acceptable salts of the GLP-1 agonist and / or pharmaceutically acceptable salts of the gastrin compound are used. The invention in particular aspects provides a pharmaceutical composition that has been adapted for administration to a subject to provide sustained beneficial effects for treating a condition and / or disease, preferably diabetes. In a modality for the prevention and / or treatment of diabetes, the composition it is in such a form that administration to a subject results in blood glucose levels that are about normal that persist in the subject for a prolonged period of time after cessation of treatment. This invention provides a conjugate comprising a GLP-1 agonist linked to or interacting with the gastrin compound wherein the interaction is, for example, by means of an amino or carboxyl group. The invention also relates to isolated covalent conjugates of the invention, and compositions comprising covalent conjugates of the invention. The GLP-1 agonist can be conjugated to species by means of an ester bond between an OH and a COOH of the gastrin compound. The conjugates of the GLP-1 agonist and the gastrin compound can be conjugated with an intermediate spacer or linker. A suitable spacer or linker can be a mono or disaccharides, an amino acid, a sulfate, a succinate, an acetate, or an oligomeric polymeric spacer or linker comprising one or more such portions. The invention also provides methods for preparing conjugates resulting in conjugates with improved pharmacokinetic properties, biological activity, and beneficial effects. The methods comprise incubating the GLP-1 agonist with a gastrin compound under conditions that allow the formation of a covalent binding between the two compounds. The invention therefore contemplates a process for preparing a covalent conjugate comprising a GLP-1 agonist linked or covalently bound to the gastrin compound, the process comprising: incubating the GLP-1 agonist with the gastrin compound under conditions and at a pH and for a sufficient time for the formation of a link or covalent ligation between the GLP-1 agonist and gastrin compound; and isolate the covalent conjugate. The above process for preparing the conjugate comprises the GLP-1 agonist and a gastrin compound which provides a conjugate with a substantial amount of the GLP-1 agonist covalently linked to the GLP-1 agonist. N-terminal or C-terminal fusion proteins or chimeric proteins, comprising a GLP-1 agonist conjugated to the gastrin compound, optionally with a spacer or linker, can also be prepared by fusing, through recombinant techniques, the sequence of terminal N or terminal C of the GLP-1 agonist and the gastrin compound sequence. The invention relates to a conjugate prepared by the process described herein. The invention also relates to the pharmaceutical formulation or composition comprising conjugates of the invention and a pharmaceutically acceptable carrier, excipient, or vehicle. The invention also relates to a formulation or pharmaceutical composition of substantially pure covalent conjugates comprising the GLP-1 agonist covalently linked to the gastrin compound which provides beneficial effects preferably sustained beneficial effects in comparison with the GLP-1 agonist alone. In one embodiment, the pharmaceutical formulation is provided consisting essentially of covalent conjugates comprising the GLP-1 agonist covalently linked without an intermediate spacer or linker to the gastrin compound. In another embodiment, the pharmaceutical formulation is provided consisting essentially of covalent conjugates comprising the GLP-1 agonist covalently linked with an intermediate spacer or linker to the gastrin compound. In aspects of the invention, the composition or conjugate comprising the GLP-1 agonist and a gastrin compound has a sustained insulinotropic activity after treatment as compared to the activity of the GLP-1 agonist or gastrin compound alone or the activity of the GLP. -1 (7-37) OH. The invention provides methods for the preparation, treatment and / or intervention of a condition and / or disease in a subject, which comprises administering the gastrin compound and the GLP-1 agonist or a pharmaceutical composition of the invention to provide the beneficial effect, in particular the sustained beneficial effect. In methods of the invention that provide effects beneficial, in particular statistically significant beneficial effects or sustained beneficial effects, the GLP-1 agonist can be selected from the group consisting of Gly8-GLP-1 (7-37), Val8GLP-1 (7-37), Val8Asp22GLP-l (7 -37), Val8Glu22GLP-I (7-37), Val8Lys22GLP-I (7-37), Val8His22GLP-I (7-37), Arg34Lys2S (N8 (? -Glu (Na-hexadecanoyl)) -GLP-1 (7 -37), Gly8-GLP-1 (7-36) amide, Val8GLP-1 (7-36) amide, Val8Asp22GLP-l (7-36) amide, Val8Glu22GLP-1 (7-36) amide, Val8Lys22GLP-l ( 7-36) amide, and Val8His22GLP-1 (7-36) amide, and the gastrin compound may comprise SEQ ID NO: 11, 12, 13 or 14, optionally associated with whey protein. invention, GLP-I (7-36) [SEQ ID NO: 5] and gastrin-17 (leu) [SEQ ID NO: 14] are administered In certain other methods of the invention, Arg34Lys26 (Ne (? -Glu ( Na-hexadecanoyl) -GLP-1 (7-37), and gastrin-17 (leu) [SEQ ID NO: 14] are administered In certain other methods of the invention, Aib8,35GLP-I (7-36) amide o It r38, Lys39'40'41'42'43'-Exendin-4 (1-39) amide and gastrin-17 (leu) [SEQ ID NO: 14] are administered. In certain additional methods of the invention, in which beneficial effects, in particular statistically significant beneficial effects or sustained beneficial effects are provided, the GLP-1 agonist is selected from the group consisting of Gly8-GLP-1 (7-37), Val8GLP -l (7-37), Val8Asp22GLP-1 (7-37), Val8Glu22GLP-1 (7-37), Val8Lys22GLP-I (7-37), Val8His22GLP-l (7-37), Arg34Lys2β (Ne (? -Glu (Na-hexadecanoyl))) -GLP-K7-37), Gly8-GLP-1 (7-36) amide, Val8GLP-l (7-36 ) amide, Val8Asp22GLP-l (7-36) amide, Val8Glu2GLP-l (7-36) amide, Val8Lys22GLP-l (7-36) amide, and Val8His22 GLP-1 (7-36) amide, and the gastrin compound comprises an amino acid sequence comprising, from the amino termini, Z-Ym-Xm-AA? -AA2 -AA3-AA4-AAs-AAs, where AAi is Tyr or Phe, AA2 is Gly, Ala, or Ser, AA3 is Trp, Val, or lie, AA4 is Met? Leu, AA5 is Asp or Glu, and AA6 is Phe or Tyr; Z is a polymer and when the polymer is a protein Z is an amino acid sequence; Ym is an optional spacer region comprising m amino acid residues of the small neutral amino acid which includes but is not limited to serine and alanine, and X is a consecutive portion of residues 1-28 of SEQ ID NO: 11 or 12, or residues 1 -17 of SEQ ID NO: 13 or 14, preferably AA? -AA2-AA3-AA4-AA5-AAS is Tyr-Gly-Trp-Met-Asp-Phe or Tyr-Gly-Trp-Leu-Asp-Phe. In a particular embodiment, Z is a whey protein, in particular human serum albumin. In one aspect, the invention provides a method for the prevention and / or intervention of a condition and / or disease discussed herein in a subject, comprising the administration of at least one GLP-1 agonist and at least one gastric compound. The GLP-1 agonist and the gastrin compound can be administered directly to a subject or brought into contact with cells (e.g. stem cells or progenitor cells) and administered to the subject. The invention also provides a combination treatment for preventing and / or treating the condition and / or disease discussed herein in a subject, comprising administering to the subject a therapeutically effective amount of at least one GLP-1 agonist and a gastrin compound. to provide beneficial effects. In one aspect, the invention provides a combination or intervention treatment that provides sustained beneficial effects after treatment. In particular, the invention provides a combination treatment for treating or preventing a condition and / or disease in a subject, comprising administering to the subject a therapeutically effective amount of at least one GLP-1 agonist and at least one gastrin compound. to produce beneficial effects, preferably sustained beneficial effects. The invention also relates to a method of treatment comprising administering a therapeutically effective amount of at least one GLP-1 agonist in combination with the administration of at least one gastrin compound which during administration to the subject with symptoms of 'diabetes produces beneficial effects, preferably sustained beneficial effects, manifested as levels of Reduced blood glucose or increased pancreatic insulin. In one aspect of the invention, the therapeutically effective amounts of the GLP-1 agonist and gastrin compound are combined prior to administration to the subject. In one embodiment, the therapeutically effective amounts of the GLP-1 agonist and the gastrin compound are mixed at a physiologically acceptable pH. In one embodiment, the invention provides a method for stimulating the proliferation of beta cells in a subject, comprising administering a therapeutically effective amount of the composition or conjugate of the invention, or administering in combination a GLP agonist and a gastrin compound. In another embodiment, the invention provides a method for increasing the number and / or size of beta cells in a subject, comprising administering a therapeutically effective amount of the composition or conjugate of the invention or administering a GLP-1 agonist in combination. and the gastrin compound. In a further embodiment, the invention provides a method for preventing or treating Type I or Type II diabetes, comprising administering a therapeutically effective amount of the composition or conjugate of the invention, or administering in combination a GLP-1 agonist and the compound of gastrin. Still in a further embodiment, the invention provides a method for alleviating the progression of the disease or obtaining a less severe stage of the disease in a person suffering from Type II diabetes, which comprises administering a therapeutically effective amount of the composition or conjugate of the invention, or administering in combination a GLP-1 agonist and the gastrin compound. The invention relates to a method for slowing the progression from decreased tolerance to glucose or Type II diabetes that does not require insulin to Type II diabetes requiring insulin, which comprises administering a therapeutically effective amount of the composition or conjugate of the invention, or administering in combination a GLP-1 agonist and the. composed of gastrin. The invention also relates to a method for increasing the synthesis capacity of insulin of a subject, comprising administering a therapeutically effective amount of the composition or conjugate of the invention, or administering in combination a GLP-1 agonist and the compound of gastrin The invention further relates to introducing islet neogenesis in a subject, comprising contacting islet precursor cells with the GLP-1 agonist and the gastrin compound, composition, or conjugate of the invention in an amount sufficient to increase the proliferation of precursor cells of islet precursor cells in a subject thereby inducing islet neogenesis. The invention contemplates a method for expanding the functional beta cell mass of pancreatic islet transplants in a diabetic patient, the method comprising administering to the patient a therapeutically effective amount of the GLP-1 agonist and the gastric compound, or a composition or conjugate of the invention . In one aspect, the invention provides methods for treating diabetes mellitus in a patient in need thereof by administering a composition comprising the gastric compound and the GLP-1 agonist in an amount sufficient to effect the differentiation of pancreatic islet precursor cells. from the patient to cells that secrete mature insulin and / or stimulated insulin synthesis in existing islet cells. The composition can be administered systemically or expressed in situ by host cells containing one or more nucleic acid constructs in an expression vector wherein the nucleic acid construct comprises the coding sequence for the gastrin compound or a sequence encoded for the agonist GLP-l or for both compounds, together with the regulatory regions of transcription and Functional translation in pancreatic islet precursor cells. The invention provides methods for treating cells, preferably cells in culture using a GLP-1 agonist and gastrin compound of the invention, or compositions, or conjugates of the invention. The invention also provides treatment methods based on cells using the GLP-1 agonist and a gastrin compound of the invention, or compositions or conjugates of the invention. See PCT / CA03 / 33595 for a description of the general culture and cell-based treatment methods. In one aspect, the invention relates to a method for expanding and differentiating stem cells or progenitor cells in insulin secreting cells comprising contacting the stem cells or progenitor cells with the GLP-1 agonist and the gastrin compound or a composition or conjugate of the invention in amounts sufficient to expand and differentiate stem cells or progenitor cells. The amount of expansion and differentiation can be significantly different compared to that which is achieved in the absence of the compounds, composition or conjugate, in particular the amount can be significantly greater compared to an amount achieved with a GLP-1 agonist or a compound of gastrin alone. In one embodiment, the stem cells or cells progenitors are contacted with the compounds, composition, or conjugate in culture. In another embodiment, the stem cells or progenitor cells are contacted with the compounds, compositions, or conjugate in the subject. The compounds, compositions or conjugates can be administered to the subject before, during, or after implantation of stem cells in the subject to expand and differentiate the cells in the subject. The cells can be obtained from pancreatic islets, umbilical cords, embryos, or stem cell lines. The method may further comprise administering an immunosuppressive agent. The invention also relates to a method for increasing the proliferation of insulin-secreting cells in culture, which comprises contacting the cells with the GLP-1 agonist and the gastrin compound, composition or conjugate of the invention in sufficient amounts to increase the proliferation of cells. The amount of proliferation can be significantly different compared to that which is achieved in the absence of the compounds, composition or conjugate. In one embodiment, the amount of proliferation is significantly greater compared to the GLP-1 agonist or the gastrin compound alone. The invention further relates to a method for supporting islet cells or precursor cells in culture, which comprises culturing the cells in the presence of an agonist GLP-1 and a gastrin compound, composition, or conjugate of the invention in an amount sufficient to sustain the cells in culture. The cells can be held in culture for a significantly longer period of time compared to cells grown in the absence of the compounds, composition or conjugate, or in the presence of the GLP agonist or the gastrin compound alone. Cells grown in the presence of the GLP-1 agonist and the gastrin compound or the composition or conjugate of the invention will be particularly useful for preparing and maintaining cells that are intended to be transplanted. In one aspect, the invention provides a method for treating a condition and / or disease, comprising administering the GLP-I agonist and the gastrin compound, the composition or conjugate of the invention with a plurality of cells to the subject in need thereof. , to thereby produce a beneficial effect, preferably a sustained beneficial effect. The method for treating the subject with a condition and / or disease described herein comprises contacting ex vivo a plurality of cells with the GLP-1 agonist and the gastrin compound, or a composition or conjugate of the invention, optionally by culturing the cells, and administer the cells to the subject who needs it. In modalities of therapeutic methods based on The aforementioned cells cells are pancreatic ductal cells and the amount of compounds / composition / conjugate used in the method is generally effective to increase the amount of cells secreting insulin in the subject. The cells can be autologous (that is, from the same subject), or they can be from another individual of the same species, or from a different species. The invention also contemplates a method of treating diabetes in a subject comprising transplanting the pancreatic islet preparation into the subject and administering a therapeutically effective amount of the GLP-1 agonist and the gastrin compound, or the composition or conjugate of the invention. In the cell-based methods of the invention, the number of cells administered to an individual affected with a condition and / or disease will vary according to the severity of the condition and / or disease, the mode of administration, and / or the site. of administration. Generally the therapeutically effective amount of the cells is a safe and effective amount, and in particular an amount necessary to provide one or more beneficial effects, in particular a sustained beneficial effect, or a synergistic effect.

The cells can be administered to subjects using a variety of means apparent to those skilled in the art. Appropriate methods include injection of the cells in a target site in a subject. The cells can be inserted into the delivery device to facilitate injection or implant in the subjects. Examples of delivery devices include tubes, eg, catheters, for injecting cells and fluids into the body of a subject. The cells can be prepared for administration in a variety of different ways. For example, the cells can be suspended in a solution or gel, or mixed with the pharmaceutically acceptable carrier, excipient or diluent in which the cells remain viable. The pharmaceutically acceptable carriers, excipients, and diluents include saline, aqueous buffer solutions, solvents and / or dispersion media. The use of such carriers and diluents is well known in the art. The solution is usually sterile, and will often be isotonic. The solution of the cells is preferably selected so that it is stable under manufacturing and storage conditions and is preserved against the contaminating action of microorganisms through the use of, for example, parabens, chlorobutanol, phenol, ascorbic acid, thimerosal, and the Similar. The modes of administration of the cells are included without intracardiac, systemic, intracoronary, intravenous, intradermal, or intra-arterial injection and injection directly into the tissues or organs at the site. proposed activity, or in proximity to the activity site. A cell preparation can be administered by any convenient route, for example by infusion or bolus injection and can be administered together with other biological active agents. The administration in some aspects is preferably systemic. A cell preparation can be administered by any convenient route, for example by infusion or bolus injection and can be administered together with other biologically active agents. The methods of the invention may further comprise measuring or monitoring one or more of the following markers: blood glucose, serum glucose, glycosylated hemoglobin in the blood, pancreatic beta cell mass, insulin in the serum, pancreatic insulin levels, beta-cell mass morphometrically determined, number of insulin-secreting cells and glucose sensitivity of insulin-secreting cells. The invention also contemplates the use of a composition comprising a combination of at least one GLP-1 agonist and at least one gastrin compound for the preparation of a medicament that provides beneficial effects, preferably sustained beneficial effects in treating a condition and / or disease. In one aspect, the invention relates to the use of a therapeutically effective amount of at least one GLP-1 agonist and at least one gastrin for the preparation of a medicament for providing beneficial effects, preferably sustained beneficial effects, in treating a condition and / or disease. In one embodiment the invention provides the use of a GLP-1 agonist and a gastrin compound for the preparation of a medicament for increasing (preferably sustained increase) the number and / or size of beta cells in a subject after treatment. In another embodiment the invention provides the use of the GLP-1 agonist and a gastrin compound for the preparation of a medicament for stimulation (preferably sustained stimulation) of beta cell proliferation after treatment. Still in a further embodiment the invention provides the use of GLP-1 and gastrin for the preparation of a medicament for the treatment of type I or type II diabetes. The invention further provides the use of a pharmaceutical composition and a conjugate of the invention in the preparation of medicament for beneficial effects, preferably sustained beneficial effects, in the treatment of conditions and / or diseases. The therapeutic efficacy and toxicity of the compounds, compositions and conjugates of the invention can be determined by standard pharmaceutical procedures in cell cultures or with experimental animals such as by calculating a statistical parameter such as the ED50 statistics (the dose that is therapeutically effective in 50% of the population) or LD50 (the lethal dose to 50% of the population). The therapeutic index is the dose ratio of therapeutic to toxic effects and this can be expressed as the ratio ED5o / LD5o. Pharmaceutical compositions which exhibit long therapeutic indices are preferred. The compounds, compositions, medicaments and conjugates of the present invention can be administered by any means which results in contacting the active agent with the action agent sites in the body of a subject or patient. The active ingredients can be administered simultaneously or sequentially, and in any order at different points in time, to provide the desired beneficial effects. The compounds, conjugates and compositions can be formulated by sustained release, for liberally, locally or systemically. These are within the ability of one of medical or veterinary skill to select a form and route of administration that optimizes the effects of the compositions, conjugates and treatments of the present invention. The compositions can be administered in oral dosage forms such as tablets, capsules (each of which includes sustained release or fixed duration release formulations), pills, powders, granules, elixirs, dyes, suspensions, syrups and emulsions. These can also be administered in intravenous (bolus or infusion), intraperitoneal, subcutaneous or intramuscular forms, all dosage forms used are known to those of ordinary skill in the pharmaceutical art. The compositions of the invention can be administered intranasally via the topical use of suitable intranasal vehicles, or via a transdermal route, for example using conventional transdermal skin patches. A dose protocol for administration using a transdermal delivery system can be continuous more than the intermittent through the dose regimen. A particular route of administration is parenteral administration, preferably peripheral parenteral administration. Parenteral administration is generally understood to refer to the injection of a dosage form into the body by a sterile syringe or mechanical device such as an infusion pump. For the purpose of the present invention parenteral routes include intravenous, intramuscular, subcutaneous and intraperitoneal routes of administration. For parenteral administration, the compounds or conjugates described herein may be combined with distilled water at an appropriate pH. The present invention includes treatments of combination that provide additives or synergistic activity, release a synergistically effective additive or amount or amount to provide a therapeutically effective amount of a GLP-1 agonist and a gastrin compound, or a conjugate or composition of the invention. Therefore, pharmaceutical compositions suitable for use in the present invention include compositions wherein the active ingredients are contained in a synergistically effective amount or a therapeutically effective amount. The dosage regimen of the invention will vary depending on known factors such as the pharmacodynamic characteristics of the agents and their mode and route of administration; the species, age, sex, health, medical condition, and weight of the patient, the nature and extent of the symptoms, the type of concurrent treatment, the frequency of treatment, the route of administration, the renal and hepatic function of the patient, and the desired effect. The effective amount of a drug required to prevent, counteract, or stop the advancement of the condition can be readily determined by an ordinarily skilled physician or veterinarian. The composition, medicament, or treatment of the invention may comprise a unit dose of at least one GLP-1 agonist and a unit dose of at least one gastric compound. The "unit dose" refers to a dose unit, that is, a single dose that is capable of being administered to a patient, and that can be handled and packaged easily, maintaining as a physically and chemically stable unit dose comprising any of the active agents as such or a mixture with one or more excipient, carriers, or solid or liquid vehicles. In one aspect, a pharmaceutical composition that is provided comprises a therapeutically effective suboptimal dose of a GLP-1 agonist and a gastrin compound that is most effective at reduced and reduced glucose levels for a sustained period followed by treatment compared to a dose of either a gastrin compound or only GLP-1 agonists. In another aspect, an improved pharmaceutical composition that is provided comprises therapeutically effective suboptimal amounts of a GLP-1 agonist and a gastrin compound in a form for chronic or acute therapy of a condition and / or disease, in particular diabetes. In one embodiment, the composition comprises a GLP-1 agonist and a gastrin compound in doses that are equal to or at least 1.1, 1.5, 2, 3, 4, 5, 6, 7, 8, 9, or 10 times lower than the doses of each of the compounds required to provide beneficial effects, preferably sustained beneficial effects, to treat a condition and / or disease. In one aspect the invention provides a pharmaceutical composition comprising between 0.5 to 6000, 100-1500, 100-6000, 1000-6000, 2000-6000- and 3000-6000 micrograms of the GLP-1 agonist per single unit and 0.5 to 6000, 100-3000, 100-6000, 1000-6000, 2000-6000 and 3000-6000 micrograms of the gastrin compound per single unit. In another aspect of the invention provides a pharmaceutical composition comprising between 0.1 to 20, 0.1 to 30, - 0.1 to 40, 0.1. up to 50 and 0.1 to 60 micrograms / kg / day of GLP-1 and 0.1 to 20, 0.1 to 30, 0.1 to 40, 0.1 to 50 and 0.1 to 60 micrograms / kg / day of the gastrin compound. A composition of the formulation of the invention can be administered to a subject continuously for 2 weeks up to 12 months, 2 weeks up to 6 months, 2-16 weeks, 2 weeks up to 12 weeks, and / or 2-8 weeks, or periodically.

In one embodiment, the ratio of the GLP-1 agonist to the gastrin compound in a composition of the invention is selected to increase the activity of the GLP-1 agonist and / or the gastrin compound and to provide beneficial effects, preferably sustained beneficial effects. . A GLP-1 agonist and a gastrin compound can be in a selected ratio to increase the activity of one or both compounds to produce beneficial effects, in particular a sustained beneficial effect, and / or to produce an additive or synergistic effect. In embodiments, the ratio of a GLP-1 agonist to a gastrin compound can be from 1: 1 to 1-.11, 1: 1 to 1: 100, 1: 1 to 1:75, 1: 1 to 1: 50, 1: 1 to 1:25, 1: 1 to 1:10, 1: 1 to 1: 5, and 1: 1. In other particular embodiments, the ratio of a gastrin compound to a GLP-1 agonist can be from 1: 1 to 1: 110, 1: 1 to 1: 100, 1: 1 to 1:75, 1: 1 to 1 : 50, 1: 1 to 1:25, 1: 1 to 1:10 and 1: 1 to 1: 5. A GLP-1 agonist can be used in combination with a gastrin compound at therapeutically effective weight ratios of between about 1: 1 to 1: 150, in particular 1: 1 to 1:50. In another embodiment, a gastrin compound can be used in combination with a GLP-1 agonist at therapeutically effective weight ratios of between about 1: 1 to 1: 150, in particular 1: 1 to 1:50. The compositions of the present invention or fractions thereof comprise pharmaceutically acceptable diluents, excipients, carriers or carriers selected based on the intended form of administration, and consistent with conventional pharmaceutical practices. The carriers, vehicles, etc. they can be adapted to provide an additive, synergistically effective or therapeutically effective amount of the active compound. The diluents, excipients, vehicles, and carriers pharmaceutically suitable are described in the standard text, Remington's Pharmaceutical Sciences, Mark Publishing Company. By way of example, for oral administration in the form of a capsule or tablet, the active components can be combined with an oral, non-toxic pharmaceutically acceptable inert carrier such as lactose, starch, sucrose, methyl cellulose, magnesium stearate, glucose, calcium. , sulfate, dicalcium phosphate, mannitol, sorbital, and the like. For oral administration in a liquid form, the drug components can be combined with an oral, non-toxic pharmaceutically acceptable inert carrier such as ethanol, glycerol, water, and the like. Suitable binders (e.g., gelatin, starch, corn sweeteners, natural sugars including glucose, synthetic and natural gums, and waxes), lubricants (e.g. sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, acetate sodium and sodium chloride), disintegrating agents (eg, starch, methyl cellulose, agar, bentonite, and xanthan gum), flavoring agents, and coloring agents can also be combined in the compositions or components thereof. In one aspect of the invention a pharmaceutical composition has a pH from about 7 to 10. Formulations for parenteral administration of a The composition of the invention may include aqueous solutions, syrups, suspensions and aqueous or oily emulsions with edible oil such as cottonseed oil, coconut oil or peanut oil. Dispersing or suspending agents that can be used for aqueous suspensions include natural or synthetic gums, such as tragacanth, alginate, acacia, dextran, sodium carboxymethylcellulose, gelatin, methylcellulose and polyvinylpyrrolidone. Compositions for parenteral administration may include sterile aqueous or non-aqueous solvents, such as water, isotonic saline, isotonic glucose solution, buffer solution, or other solvents conveniently used for parenteral administration of therapeutically active agents. A composition intended for parenteral administration may also include conventional additives such as stabilizers, buffers, or preservatives, for example antioxidants such as methylhydroxybenzoate or similar additives. In one embodiment, a solid form pharmaceutical composition is provided (eg, tablets, capsules, powder or powdered form) comprising a crystalline or amorphous GLP-1 agonist and a crystalline or amorphous gastrin compound. In another embodiment, the invention relates to a liquid drug formulation comprising pharmaceutically acceptable salts of a GLP-1 agonist and a gastrin compound, and lyophilized drugs that can be reconstituted to provide suspensions that are stable and suitable for parenteral administration. In a particular embodiment, the invention relates to an aqueous composition comprising pharmaceutically acceptable salts of a GLP-1 agonist and a gastrin compound, and a solvent system having solubilization effects. The invention also provides a drug comprising an aqueous formulation of pharmaceutically acceptable salts of a GLP-1 agonist and a gastrin compound with at least one solubilizer. A composition of the invention can also be sterilized by, for example, filtration through a filter that maintains the bacteria, in addition to sterilizing agents to the composition, irradiation of the composition, or heat to the composition. Alternatively, the compounds, conjugates, and compositions of the present invention can be provided as sterile solid preparations, for example lyophilized powder, which are easily dissolved in a sterile solvent immediately prior to use. In addition to the formulations described herein, the compositions may also be formulated as a stored preparation. Such action formulations Prolonged can be administered by implantation (eg, subcutaneously or intramuscularly) or by intramuscular injection. Thus, for example, the fractions can be formulated with suitable hydrophobic or polymeric materials (for example, as an emulsion in an acceptable oil), or ion exchange resins, or as sparing soluble derivatives, for example, as a sparingly soluble salt. The compositions of the invention and components thereof may comprise soluble polymers as carriers of the target drug. After the pharmaceutical compositions have been prepared, they can be placed in an appropriate container and labeled for treatment of an indicated condition. For administration of a composition of the invention, such labels include amount, frequency and method of administration. The present invention also includes methods for using the compositions of the invention in combination with one or more additional therapeutic agents including without limitation immunosuppressive agents, anti-obesity agents, antidiabetic agents, appetite regulating drugs, antihypertensive agents, agents for treatment and / or prevention of complications resulting from or associated with a condition and / or disease, in particular diabetes and obesity, anti-nausea, anti-headache medications, and general medications to treat or prevent side effects. Since the present invention relates to a method of treatment which comprises a combination of active agents that can be administered separately or as conjugates, the invention also provides a kit comprising a GLP-1 agonist and a gastrin compound, a pharmaceutical composition or conjugated in kit form. The invention also relates to a pharmaceutical kit comprising a bottle with a GLP-1 agonist and another bottle with a bottle of gastrin in a box. A kit can comprise a package which are houses, a container containing a conjugate or composition of the invention and also has home instructions for administering the conjugate or composition to a subject. The invention will be described in more detail by way of specific examples. The following examples are entirely for illustrative purposes, and are not intended to limit the invention in any way. Those of ordinary skill in the art will readily recognize a variety of non-critical parameters that can be changed or modified to essentially provide the same result.

Example 1 Effects of gastrin in combination with GLP-1 (Bache GLP-1 (7-36) Amida, Human) in NOD mice with diabetic acuity. This example shows methods and compositions for reversing diabetes in diabetic NOD mice by stimulating β-cell neogenesis in vivo after systemic treatment with GLP-1 and gastrin. The_ NOD female mice aged 12-16 weeks are treated for 18 days only with vehicle (PBS), GLP-l (300 μg / kg / day), or GLP-l (3007g / kg / day) + gastrin (3 μg / kg / day), by injection intraperitoneally 2 times daily within 2 days after the onset of diabetes. The onset of diabetes is determined by fasting blood glucose (FBG) levels (9-15 mM compared to normal FBG <6.0 mM). The mice were monitored daily by urine glucose and weekly by levels of FBG. At the beginning of the treatment, the fasting blood glucose levels are in the range between 11-14 mM. After 18 days of treatment, FBG was 24 + 1 mM in vehicle treated mice, 13 + 2 mM in mice treated with GLP-1 alone, and 6 ± 1 M in mice treated with the combination of GLP-I and gastrin ( mean ± SE, n = 4 mice in treated groups, n = 6 mice in the control group). After 18 days treatment therapy was stopped and FBG monitored weekly for 6 weeks additional One week after completing treatment, FBG levels returned to normal in mice injected with the combination therapy, and the remaining of such levels throughout the end of the study at 6 weeks after treatment. Comparatively, the untreated group of animals has been sacrificed due to the severity of the disease after 5 weeks. Mice treated only with GLP-1 show transient improvement in FBG levels up to 2 weeks after the treatment was stopped, after the fasting blood glucose levels progressively increased and are similar to the levels observed in the untreated vehicle group by the end of the study. The results of the study are illustrated in figure 1. These results showed that a short course of a combined treatment of gastrin and GLP-1 for NOD mice • Diabetics normalizes hyperglycemia to effectively treat diabetes, and has a prolonged effect on fasting blood glucose levels which indicates a stimulation of beta cell neogenesis and insulin production.

Example 2 Effects of gastrin (Gl) in combination with GLP-1 in mice NOD with diabetic acuity. Objective: NOD mice spontaneously develop insulin-dependent diabetes as a result of autoimmune destruction of pancreatic islet β cells. This study is aimed at correcting diabetes in NOD mice by regenerating islet β cells using GLP-1 and gastrin (Gl).

Method: Female NOD mice aged 12-16 weeks were treated for 18 days only, with vehicle (PBS) or with 300 μg / kg / day of GLP-1 in combination with 3 μg / kg / day of gastrin (Gl ) by intraperitoneal injection (ip). The animals were injected for 18 days, twice daily, within 2 to 5 days after the onset of diabetes. Fasting blood glucose levels (FBG) are 9-15 mM at the onset of diabetes (normal FBG <6.0 mM). The mice were monitored daily for glucose levels in the urine and weekly for FBG levels during the treatment, and for an additional 6 weeks after the treatment was stopped. Pancreatic insulin levels are determined in each group as well as histological analysis of the pancreatic tissue were carried out. The Pancreatic tissues were fixed and stained for cells that produce insulin. The beta cell mass is determined by morphometric analysis.

Results: After 18 days of daily treatment, in animals treated with the combination of 30 μg / kg / day of GLP-1 and 3 μg / kg / day of Gl, the fasting blood glucose was 6.1 ± 0.7 mM, while fasting blood glucose was 24.4 ± 1.5 mM in the vehicle-treated group. In comparison, animals treated with GLP-I only have fasting blood glucose levels of 12.5 ± 2.2 mM. These data indicate that the combination treatment of GLP-1 and gastrin are more effective than GLP-1 alone in glucose levels controlled in NOD mice. All treatments were stopped after 18 days and FBG was monitored weekly for an additional 6 weeks. One week after completing the treatment, the levels of FBG are normalized (under 6 mM) in mice injected with the combination therapy and the rest of such levels throughout the end of the study with FBG of 4.3 ± 0.2 mM at 6 weeks after treatment. Comparatively, the animals treated with GLP-l reached maximum blood glucose levels (above 30 mM) and suffer from diabetic complications. The untreated group of animals had to sacrifice due to the severity of the disease after 5 weeks after treatment (figure 1 and figure 2). These data indicate that the combination of GLP-1 and gastrin are effective in restoring normal blood glucose levels even after 6 weeks after treatment, while animals treated with GLP-1 develop severe hyperglycemia. Figure 3 shows that non-diabetic animals have approximately 10 μg of insulin per pancreas, while diabetic animals with high glucose levels have 0.5 to 1.0 μg of pancreatic insulin. These data show that NOD mice require less than 10% of their pancreatic insulin to regulate glucose levels. 5 weeks after the onset of diabetes, untreated animals have minimal levels of pancreatic insulin, and at this stage the animal has glucose levels of 30-32 mM and suffers from diabetic complications. The group treated with GLP-1 has pancreatic insulin levels of 1.0 to 1.5 μg, which are higher than in untreated animals, suggesting that GLP-1 stimulates some islet cell in the model of NOD mice. Surprisingly, the animals treated with GLP-1 and gastrin had about 8 μg per pancreas which is not only significantly greater than GLP-1 but over 80% of normal non-diabetic pancreatic insulin levels. These Studies show that a treatment in combination with GLP-1 and gastrin is very robust in stimulating islet cell regeneration that is capable of reversing the disease for long periods of time after treatment. GLP-1 and gastrin are able to repair the pancreatic insulin content of the lower levels measured after the onset of diabetes and post-treatment for a similar level to be measured in normoglycemic mice. Correction of hyperglycemia in NOD mice was significantly correlated with the increase in pancreatic insulin content (r = 0.90), as presented in figure 4. Figure 5 shows that cells stained with insulin (in dark brown), are few in mice? OD with diabetic acuity before treatment. The number of these islet cells also decreases in the untreated group for a time. Histological examination revealed intensely insulin-stained islets, large adjacent to the pancreatic ducts and surrounding but not invaded by mononuclear leukocytes in mice treated with GLP-1 and gastrin. The beta cell mass decreases from 0.41 mg to 0.01 mg during the course of the experiment (8 weeks), while the beta cell mass increases to 1.05 mg in the group of animals treated with GLP-I and gastrin. The mass of beta cells in animals does not Diabetics have been reported to be in the range of 1.0-1.5 mg. Treatment with GLP-1 and gastrin significantly increases the beta cell mass in NOD mice for almost normal levels, even when examined 6 weeks after treatment. Figure 6 demonstrates the dyeing of islet cells from the pancreatic duct in NOD mice treated with vehicle and GLP-1 and gastrin. The data demonstrate that the beta cell mass of the islet cell group in the pancreatic ducts decreased from 0.06 to 0.01 during the course of the experiment (8 weeks), whereas these groups of beta cell mass are increased up to 0.17 mg in mice NOD treated with GLP-1 and gastrin. These data indicate that treatment with GLP-1 and gastrin induces islet neogenesis in mice involved in islet cell precursors in the pancreatic duct. In summary, these studies show that the treatment With GLP-1 and gastrin induces the regeneration of islet cells in the model of mice, OD sufficiently to unbalance the destruction that can be followed in these disease models, which result in net accumulation of islet cells in the pancreas.

Conclusion: A short course of treatment with GLP-1 and gastrin in mice? Diabetic OD normalizes hyperglycemia and has a 0 prolonged effect on fasting blood glucose levels for periods of at least 6 weeks after treatment. In addition, the data show that GLP-1 and gastrin is capable of stimulating pancreatic insulin levels of approximately 80-90% of normal levels, while GLP-1 alone has a modest effect only. Additionally, histological analysis of the pancreas shows that islet cells appear normal and with large numbers of insulin-producing cells, despite being surrounded by inflammatory cells. The morphometric analysis of the pancreas shows that treatment with GLP-1 and gastrin increases the mass of beta cells in the pancreas, and shows signs of inducing neogenesis by increasing the mass of beta cells in the pancreatic ducts. The treatment with GLP-1 and gastrin is a potent inducer of islet cell regeneration that can restore normal levels of glucose and pancreatic insulin in the NOD mouse model.

Example 3 Modified gastrin compounds / conjugates of PCT / CA03 / 01778 in combination with GLP-1 in the prevention of diabetes progression in NOD mice with recent onset of diabetes. The effect of the treatment by a combination of GLP-1 and Unmodified gastrin and modified gastrin / GLP-1 compounds / conjugates will be examined in NOD mice with recent onset of diabetes, to determine whether the administration of both GLP-1 and gastrin prevents severe hyperglycemia as well as an increase in insulin content pancreatic in NOD mice with diabetes of recent onset. GLP-1 for use in the biologically active fragment of GLP-1 from human / mouse GLP-1 (having residues at positions 7-36 compared to the precursor from which the fragment is processed, obtained from Bachem H6795 ). The compounds / gastrin conjugates to be used are as follows: Compound B-gastrin as human synthetic gastrin I having 17 amino acid residues with a Leu residue at amino acid position 15, Compound E-gastrin as human synthetic gastrin I having 2 -17 amino acid residues. Compound Q, gastrin as human synthetic gastrin I having 2-17 amino acid residues with a HSA polymer bound by (GA) 5 (ie, Gly-Ala-Gly-Ala-Gly-Ala-Gly-Ala-Gly) -To) . Non-obese diabetic female mice (NOD), ages 12-14 weeks, will be observed for the onset of diabetes onset (fasting blood glucose> 8.0 to 15 tpmol / 1), and within 48 hours after the onset of symptoms, four groups of mice each will be treated as follows: a group will be treated only with vehicle; and the other group will be will administer 100 μg / kg / day of GLP-1, and the remaining groups will be treated with a combination of GLP-1 (100 μg / kg / day) and gastrin compound (3 μg / kg / day equivalent), each treatment given by intraperitoneal route daily. The therapy will be administered for 14 days up to 18 days. The animals will be observed weekly for fasting glucose levels (FBG). FBG levels will be measured at about 12 hours after the food has been removed, and 24 hours after the last injection of peptide or vehicle. With the suspension of therapy, all mice will be observed for FBG levels during the next 4 weeks (weeks 2-6) in order to determine whether the prevention of hyperglycemia persisted after the completion of therapeutic treatment. Treatment will stop at 14 days until 18 days. The protocol includes sampling these mice for data again at 6 weeks, and blood collecting the blood for the FBG and C-peptide assay in plasma, and sacrificing the mice for pancreatic insulin determinations and inflammation log of islets (insulitis). From the beginning of the treatment, the mice will not receive treatment with insulin replacement or immunosuppression. The following parameters will be evaluated: survival rates, pancreatic insulin levels, presence of islet inflammation and fasting blood glucose levels.

GLP-I in combination with modified compounds / conjugates of gastrin (Compound E or Q) with more extended half-lives can provide improved reduction of blood glucose levels in diabetic animals.

Example 4 - By using the standard Fmoc synthesis, two different "reactive" gastrin compounds will be produced: compound A is a modified gastrin 17 peptide having an additional cysteine at the N-terminal end; Compound B is a modified gastrin-17 peptide having 10 additional amino acids of alternating glycine and alanine (5 amino acids each) as a spacer region with an additional cysteine at the end of the N-terminus. Non-obese diabetic female mice will be observed (NOD) for the development of diabetes (determined to be fasting blood glucose, FBG level greater than 6.6 mmol / 1) and with the onset of diabetes, divided into four groups. The mice will be treated either with vehicle as control; or with gastrin 17, with compound A or with compound B (same molar concentration of active ingredient, that is, gastrin, to be used for all three groups treated with gastrin), administered by intraperitoneal (ip) injection once daily for 14 days.

Fasting blood glucose (FBG) and pancreatic insulin levels will be measured and determined. In addition, the serum half-life of gastrin will be measured as well as circulating serum levels of gastrin. It is anticipated that of the three groups treated with gastrin, both groups of NOD mice that are treated with either compound A or compound B will maintain higher circulating levels of serum gastrin. Furthermore, the measured half-life of gastrin will be higher in mice treated with either compound A or compound B compared to unmodified gastrin. In addition, it is also anticipated that when comparing the control group treated with vehicle which registers increasingly high levels of FBG, all three groups of treated animals will have decreased levels of FBG. Animals treated with either Compound A or Compound B may have even lower levels of FBG as well as increased levels of pancreatic insulin compared to animals treated with unmodified gastrin. The present invention is not to be limited in scope by the specific embodiments described herein, since such embodiments are intended to be but simple illustrations of one aspect of the invention, and any functionally equivalent embodiments are within the scope of the invention. In fact, various modifications of the invention in addition to those shown and described herein for those skilled in the art from the foregoing description and appended drawings. It is intended that such modifications fall within the scope of the appended claims. All publications, patents and patent applications referred to herein are incorporated by reference in their entirety to the same degree as if each publication, patent or individual patent application is specifically and individually indicated to be incorporated by reference in its entirety. The citation of any reference herein is not an admission that such a reference is available as a prior art of the current invention.

Table 1 GLP-l Agonist Source Novo Nordisk GLP-l analog derivatives with a lipophilic WO 98/08871 substituent (protraid action profile) Fragment of GLP-l as The General Hospital insulinotropic hormone Corporation WO 87/06941 GLP-1 derivatives with The General Hospital insulinotropic activity Corporation WO 90/11296 GLP-1 analogues showing Buckley et al. improved stability or WO 91/11457 improved ability to stimulate insulin production Analogs and derivatives of GLP-1 Eli Lilly & Co. (stimulate secretion or EP 0708179-A2 insulin biosynthesis in poorly functioning beta cells) GLP-l agonist Source GLP-l associated molecules Eli Lilly and Company with a metal cation 6,133,235 divalent 5,977,071 Theratech, Inc. oral delivery systems with GLP-l 5,863,555 GLP-l analogs Eli Lilly and Company 5,981,488 GLP-imitations l Bristol-Myers Squibb Company WO 03/033671 GLP-1 long-life Conj uchem, Inc. US 6,593,295 US 6,514,500 US 6,329,336 GLP-l precursor Genzyme Corporation WO 03/014318 GLP-l complexes Eli Lilly and Company 6,358,924 Modified peptides Theratechnologies Inc. WO 02/10195 GLP-1 and related molecules Zealand Pharma A / S WO 2004/005342 It is noted that in relation to this date, the best method known to the applicant to carry out the aforementioned invention, is that which is clear from the present description of the invention.

Claims

Claims Having described the invention as above, the claim contained in the following claims 1 is claimed as property. A pharmaceutical composition characterized in that it comprises a GLP-1 agonist and a gastrin compound that provides beneficial effects in relation to each compound alone, and optionally a pharmaceutically acceptable carrier, excipient or vehicle.
2. The pharmaceutical composition according to claim 1, characterized in that it is in a form that provides normal levels of glucose in the blood in a subject that persists for a prolonged period of time after administration.
3. The pharmaceutical composition according to any of the preceding claims, characterized in that it comprises therapeutically effective amounts of a GLP-1 agonist and a gastrin compound in a form for chronic or acute therapy of a subject in need thereof.
4. The pharmaceutical composition according to claim 3, characterized in that the amounts Therapeutically effective are sub-optimal in relation to the amount of each compound administered alone for the treatment of diabetes.
5. The pharmaceutical composition according to any of the preceding claims, characterized in that the ratio of the GLP-1 agonist to the gastrin compound is selected to increase the activity of the GLP-1 agonist or gastrin compound.
6. The pharmaceutical composition according to claim 1, characterized in that the ratio of the GLP-1 agonist to the gastrin compound is from about 1: 1 to 1: 110, 1: 1 to 1: 100, 1: 1 to 1: 75, 1: 1 at 1:50, 1: 1 at 1:25, 1: 1 at 1:10, 1: 1 at 1: 5, and 1: 1.
7. The pharmaceutical composition according to claim 1, characterized in that the ratio of a gastrin compound to the GLP-1 agonist is from about 1: 1 to 1: 110, 1: 1 to 1: 100, 1: 1 to 1 : 75, 1: 1 to 1:50, 1: 1 to 1:25, 1: 1 to 1:10, and 1: 1 to 1: 5.
8. The pharmaceutical composition according to any of the preceding claims, characterized in that the GLP-1 agonist is used in combination with the gastrin compound at therapeutically effective weight ratios of between about 1: 1.5 to 1: 150, preferably 1: 2 to 1:50.
9. The pharmaceutical composition according to any of the preceding claims characterized in that the GLP-1 agonist and the gastrin compound are present in doses that are at least about 1.1 to 1.4, 1.5, 2, 3, 4, 5, 6, 7 , 8, 9, or 10 times less than the doses of each compound only required to treat a condition and / or disease.
10. The pharmaceutical composition according to claim 1, characterized in that it comprises an additive amount of the GLP-1 agonist and the gastrin compound in a pharmaceutically acceptable excipient, carrier or vehicle.
11. The pharmaceutical composition according to claim 1, characterized in that it comprises a synergistically effective amount of the GLP-1 agonist and the gastrin compound in a pharmaceutically acceptable excipient, carrier or vehicle.
12. The pharmaceutical composition according to claim 1, characterized in that it comprises between 0.1 to 20, 0.1 to 30, 0.1 to 40, 0.1 to 50, and 0.1 to 60 micrograms / kg / day of GLP-1 agonist and 0.1 to 20, 0.1 to 30, 0.1 to 40, 0.1 to 50, and 0.1 to 60 micrograms / kg / day of gastrin compound.
13. The pharmaceutical composition according to claim 2, characterized in that the beneficial effects are one or more of the following: reduced or absent inflammation of islets, decreased progression of the disease, increased survival, or decreased symptoms of a disease or condition.
14. The pharmaceutical composition according to any of the preceding claims, characterized in that the beneficial effects are sustained beneficial effects that persist for a prolonged period of time after the termination of the treatment.
15. The pharmaceutical composition according to claim 14, characterized in that the beneficial effects are sustained for at least about 2, 4, 5, 6, or 10 weeks, 2 to 4 weeks, 2 to 8 weeks, 2 to 12 weeks, 2 at 24 weeks, 2 weeks at 12 months, and 2 weeks at 18 months after treatment.
16. The pharmaceutical composition according to claim 15, characterized in that the sustained beneficial effects may be manifested as an increased production of the C-peptide, increased production of pancreatic insulin, and low or near normal blood glucose levels for a prolonged period after of the treatment.
17. The pharmaceutical composition according to any of the preceding claims, characterized in that the beneficial effect is at least about 0.5%, 1%, 2%, 5%, 10%, 15%, 20%, "30%, 33% , 35%, 40%, 45%, or 50% of 'increase in pancreatic insulin levels.
18. The pharmaceutical composition according to any of the preceding claims, characterized in that the beneficial effect is at least about 2%, 5%, 10%, 15%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, or 90% decrease in blood glucose levels.
19. The pharmaceutical composition according to any of the preceding claims, characterized in that the beneficial effect is a decrease in blood glucose levels for a period of at least 2, 4, 6, 8, or 10 weeks, 2 to 4 weeks. , 2 to 6 weeks, 2 to 8 weeks, 2 to 12 weeks, 2 to 24 weeks, 2 weeks to 12 months, and 2 weeks to 18 months after treatment.
20. The pharmaceutical composition according to any of the preceding claims, characterized in that the GLP-1 agonist is a GLP-1 (1-37), GLP-1 (7-36) amide, fragments, analogues, and derivatives thereof, and active metabolites and prodrugs of GLP-1.
21. The pharmaceutical composition according to any of the preceding claims, characterized in that the GLP-1 is GLP-1 (7-36) of SEQ ID NO 5 or gastrin 17 (leu) of SEQ ID NO. 14
22. The pharmaceutical composition according to any of the preceding claims, characterized in that the GLP-1 agonist comprises a precursor polypeptide of the formula GLP-1 (7-R) wherein R is 36, 37, 38, 39, 40, 41, 42, 43, 44, and 45, and wherein optionally up to 5, 10, or 15 amino acid residues are replaced with any residue of α-amino acids.
23. The pharmaceutical composition according to any of the preceding claims, characterized in that the GLP-1 agonist is an analog or derivative of GLP-1 listed in Table 1.
24. The pharmaceutical composition according to any of the preceding claims, characterized in that the gastrin compound is gastrin 71 [SEQ ID NO. 15], gastrin 52 [SEQ ID NO. 16], gastrin 34 (large gastrin) [SEQ ID NO. 11 or 12], gastrin 17 (small gastrin) [SEQ ID NO. 13 or 14], gastrin 14 [SEQ ID? O. 17], gastrin 8, gastrin 6 [SEQ ID? O. 18 or 19], pentagastrin, and tetragastrin.
25. The pharmaceutical composition according to any of the preceding claims, characterized in that the gastrin compound is a compound of the formula Z-Ym-Xn-AA1-AA2-AA3-AA4-AA5-AA6, wherein AA1 is Tyr or Phe, AA2 is Gly, Ala, or Ser, AA3 is Trp, Val, or lie, AA4 is Met or Leu, AA5 is Asp or Glu, and AA6 is Phe or Tyr which is optionally amidated; Z is a carrier, preferably a polymer, more preferably a protein; Ym is an optional spacer region comprising m amino acid residues of a neutral small amino acid including but not limited to serine and alanine, and X is any consecutive porcine of residues 1-28 of SEQ ID NO: 11 or 12, or residues 1-11 of SEQ ID? O. 13 or 14, preferably AA1-AA2-AA3-AA4-AA5-AA6 is Tyr-Gly-Trp-Met-Asp-Phe or Tyr-Gly-Trp-Leu-Asp-Phe.
26. A pharmaceutical composition according to any preceding claim characterized in that the GLP-1 agonist is selected from the group consisting of Gly8-GLP-1 (7-37), - Val8GLP-I (7-37), Val8Asp22GLP-I (7- 37), Val8Glu22GLP-I (7-37), Val8Lys22GLP-I (7-37), Val8His22GLP-I (7-37), Arg34Lys26 (Ne (g-Glu (Na-hexadecanoyl))) -GLP-I (7-37), Gly8-GLP-1 (7-36) mida, Val8GLP-I (7-36) amide, Val8Asp22GLP-l (7-36) amide, Val8Glu22GLP-I (7-36) mida, Val8Lys22GLP-I (7-36) amide, and Val8His22GLP-I (7-36) amide, and the gastrin compound is gastrin comprising SEQ ID NO. . 11, 12, 13, 14, 17, or 18.
27. The pharmaceutical composition according to any preceding claim characterized in that the GLP-1 agonist is Arg34Lys26 (Ne (g-Glu (Na-hexadecanoyl))) -GLP-I (7-37) and the gastrin compound is 15Leu gastrin 17 [ SEQ ID NO. 14].
28. The pharmaceutical composition according to claim 27 characterized in that the gastrin compound is associated with a whey protein, preferably human serum albumin.
29. A method for the preparation of a stable pharmaceutical composition of a GLP-1 agonist, characterized in that it comprises mixing a GLP-1 agonist, a gastrin compound, and a pharmaceutically acceptable carrier, excipient or vehicle to physically stabilize the GLP agonist -l and adapted to provide beneficial effects, preferably sustained beneficial effects.
30. A conjugate, characterized in that it comprises a GLP-1 agonist linked to a gastrin compound to provide beneficial effects, in particular sustained beneficial effects.
31. The conjugate in accordance with the claim 30, characterized in that the GLP-1 agonist is selected from the group consisting of Gly8-GLP-1 (7-37), Val8GLP-I (7-37), Val8Asp22GLP-l (7-37), Val8Glu22GLP-l (7-37), Val8Lys22GLP-l (7-37), Val8His22GLP-l (7-37), Arg34Lys26 (Ne (g-Glu (? A-hexadecanoyl))) -GLP-l (7-37), Gly8-GLP-1 (7-36) amide, Val8GLP-l (7-36) amide, Val8Asp22GLP-I (7-36) amide, Val8Glu22GLP-I (7-36) amide, Val8Lys22GLP-I (7-36) amide, and Val8His22GLP-I (7-36) amide, and the gastrin compound is gastrin comprising SEQ ID NO. 11, 12, 13, 14, 17, or 18 optionally associated with a whey protein.
32. A method for the treatment or prevention of a condition and / or disease in a subject, characterized by comprising administering to a subject a therapeutically effective amount of a GLP-1 agonist and a gastrin compound, or a composition or conjugate in accordance with any preceding claim, to produce a sustained beneficial effect.
33. The method according to claim 32, characterized in that the sustained beneficial effect is a decrease in blood glucose levels for a period of at least 2, 4, 6, 8, or 10 weeks, 2 to 4 weeks, 2 to 6 weeks, 2 to 8 weeks, 2 to 12 weeks, 2 to 24 weeks, 2 weeks to 12 months, and 2 weeks to 18 months after treatment.
34. A method of treatment, characterized in that it comprises administering to a subject a therapeutically effective amount of at least one GLP-1 agonist in combination with the administration of at least one gastrin compound which upon administration to a subject with symptoms of diabetes provides beneficial sustained.
35. The method according to claim 34, characterized in that administration with at least one GLP-1 agonist in combination with the administration of at least one gastrin compound provides sustained beneficial effects of at least one diabetes symptom.
36. The method according to claim 34 or 35, characterized in that therapeutically effective amounts of the GLP-1 agonist and the gastrin compound are combined prior to administration to the subject.
37. The method according to claim 34 or 35, characterized in that therapeutically effective amounts of the GLP-1 agonist and the gastrin compound are sequentially administered to the subject.
38. The method according to any preceding claim characterized in that the therapeutically effective amounts of a GLP-1 agonist and a gastrin compound are synergistically effective amounts.
39. A method of preparing a stable pharmaceutical composition of a GLP-1 agonist, characterized in that it comprises mixing a GLP-1 agonist, a gastrin compound, and a carrier, excipient or vehicle pharmaceutically acceptable to physically stabilize the GLP-1 agonist and adapted to provide beneficial effects preferably sustained beneficial effects.
40. A method for the treatment of a condition and / or disease, characterized in that it comprises administering a GLP-1 agonist and a gastrin compound, or a composition or conjugate according to any preceding claim with a plurality of cells to a subject in need of same to thereby produce beneficial effects, preferably sustained beneficial effects.
41. The method according to any preceding claim, characterized in that the condition and / or disease is dyslipidemia, hyperglycemia, severe hypoglycemic episodes, apoplexy, left ventricular hypertrophy, arrhythmia, bacteremia, septicemia, irritable bowel syndrome, respiratory distress syndrome, functional dyspepsia , diabetes, catabolic changes after surgery, hyperglycemia induced by tension, gastric ulcers, myocardial infarction, impaired glucose tolerance, hypertension, Alzheimer's disease and other central and peripheral neurodegenerative conditions, chronic heart failure, retention states of fluids, metabolic syndrome and related diseases and disorders, and obesity.
42. The method according to claim 40 or 41 characterized in that the condition and / or disease is diabetes.
43. A method for inducing islet neogenesis in a subject, characterized in that it comprises contacting the islet precursor cells with a GLP-1 'agonist and a gastrin compound, or a composition, or conjugate according to any preceding claim in an amount sufficient to increase the proliferation of islet precursor cells in the subject, whereby islet neogenesis is induced.
44. A method for expanding and differentiating stem cells in insulin secreting cells, characterized in that it comprises contacting the stem cells with an effective amount of a GLP-1 agonist and a gastrin compound or a composition or conjugate according to any preceding claim.
45. The method according to any preceding claim characterized in that the agonist GLP-l is selected from the group consisting of Gly8-GLP-1 (7-37), Val8GLP-I (7-37), Val8Asp22GLP-I (7-37), Val8Glu22GLP-I (7-37), Val8Lys22GLP- l (7-37), Val8His22GLP-I (7-37), Arg34Lys26 (Ne (g-Glu (Na-hexadecanoyl))) -GLP-I (7-37), Gly8-GLP-1 (7-36) amide, Val8GLP-l (7-36) amide, Val8Asp22GLP-l (7-36) amide, Val8Glu22GLP-l (7-36) amide, Val8Lys22GLP-l (7-36) amide, and Val8His22 GLP-l (7- 36) amide, and the gastrin compound is gastrin comprising SEQ ID NO. 11, 12, 13, 14, 17, or 18 associated with a whey protein.
46. The method according to any preceding claim, characterized in that the GLP-1 agonist is Arg34Lys26 (Ne (g-Glu (Na-hexadecanoyl))) -GLP-1 (7-37) and the gastrin compound is 15Leu gastrin 17 [ SEQ ID NO. 14].
47. The use of a composition comprising a combination of at least one GLP-1 agonist and at least one gastrin compound for the preparation of a medicament for the treatment of a condition and / or disease.
48. The use of a GLP-1 agonist for the manufacture of a medicament for the treatment of a condition and / or disease to be used in combination with a gastrin compound.
49. The use according to claim 45 or 46 wherein the GLP-1 agonist is "selected from the group consisting of Gly8-GLP-1 (7-37), Val8GLP-l (7-37), Val8Asp22GLP-l (7 -37), Val8Glu22GLP-I (7-37), Val8Lys22GLP-I (7-37), Val8His22 GLP-I (7-37), Arg34Lys26 (Ne (g-Glu (Na-hexadecanoyl))) -GLP-I (7-37), Gly8-GLP-1 (7-36) amide, Val8GLP-I (7-36) amide, Val8Asp22GLP-l (7-36) amide, Val8Glu22GLP-l (7-36) amide, Val8Lys22GLP-l (7-36) amide, and Val8His22 GLP-l (7-36) amide, and the gastrin compound is gastrin comprising SEQ ID NO. 11, 12, 13, 14, 17, or 18.
50. The use according to any preceding claim wherein the GLP-1 agonist is Arg34Lys26 (Ne (g-Glu (Na-hexadecanoyl))) -GLP-I (7-37) and the gastrin compound is 15Leu gastrin 17 [SEQ. ID NO. 14].
51. The use according to any claim wherein the condition and / or disease is dyslipidemia, hyperglycemia, severe hypoglycemic episodes, stroke, left ventricular hypertrophy, arrhythmia, bacteremia, septicemia, irritable bowel syndrome, functional dyspepsia, respiratory distress syndrome, diabetes , catabolic changes after surgery, hyperglycemia induced by tension, gastric ulcers, myocardial infarction, impaired glucose tolerance, hypertension, Alzheimer's disease and other central and peripheral neurodegenerative conditions chronic heart failure, fluid retention states, metabolic syndrome and related diseases and disorders, and obesity.
52. A kit form characterized in that it comes from a composition or conjugate according to any preceding claim.