CN1729016A - Treatment of diabetes - Google Patents

Abstract

Compositions and methods are provided for islet neogenesis therapy comprising a member of a group of factors that complement a gastrin/CCK receptor ligand, with formulations, devices and methods for sustained release delivery and for local delivery to target organs.

Description

The treatment of diabetes

Invention field

The present invention relates to the method and composition with pancreas islet regeneration therapy and immunosuppressant treatment diabetic subjects, be used for preparation and the method for local conveying and composition sustained release.

Background of invention

The serious type of common disease diabetes is because due to the shortage or relative deficiency of pancreatic beta cell insulin secretion. So diabetes rely on the injection exogenous insulin with high blood-glucose (hyperglycaemia) complication of preventing damage life. Unless the experimenter depends on very high glucose test and the scheme of insulinize (intensive insulin therapy), otherwise insulinize can not prevent the chronic long complication of the organ injury that hyperglycaemia causes. Intensive insulin therapy has increased owing to the excessive acute risk of hypoglycemia that causes of insulin, with being fatal acute and serious state of consciousness change.

An about million people suffers from juvenile form or type i diabetes in U.S. population, about 30,000 new cases of annual appearance. In addition, on the level of epidemic disease ratio, type ii diabetes (being also referred to as grow up morbidity or insulin-resistant diabetes) experimenter's quantity is extremely huge and increase sharply, and a kind of pancreas that causes exhausts disease with insufficient insulin.

Unusual high blood-glucose (hyperglycaemia) is the characteristics of diabetes, does not treat if let alone, and will cause various pathological states, for example, incurable peripheral vascular ulcer, causes retinal damage and the kidney failure of losing one's sight. I type and type ii diabetes are replied the blood glucose levels generation of measuring by experimenter's glucose self monitoring all with the insulin injection treatment, need insulinize although not all II type experimenter can proceed to. Typically, carry the insulin of the many multiple doses of experimenter every day. The serious pathological examination of diabetes is strictly not relevant with blood glucose levels control.

May treating of diabetes will be to recover the β cell function in order to the variation generation of blood glucose levels is replied and the release of dynamic adjustments insulin. This can finish by transplantation of pancreas, but this method typically is limited to the diabetes that kidney failure needs kidney transplant. Similarly, transplantation of pancreas may need semelincident immunization to suppress to prevent the autoimmunity destruction of allograft refection and pancreas graft.

Recently, the graft of people's pancreas islet goods of separation has successfully been reversed the insulin diabetes for a long time in the human experimenter. Yet, concerning every receptor, need a large amount of donor islet cell materials, and the supply of islet cells material can not be enough to satisfy the demand.

General introduction

Characteristic of the present invention is the method for the treatment of diabetes, the method comprises the composition that comprises gastrin/cck receptor part and the factor (FACGINT) of replenishing the gastrin that is used for the pancreas islet regenerative therapy to administration treatment effective dose, as long as FACGINT is not the EGF receptors ligand.

Such as term used herein, FACGINT is selected from least a in lower group: the glucogan-like peptide 1 receptor part; The glucagon-like peptide 2 receptors ligand; Gastrointestinal inhibitory peptide (GIP) receptors ligand; Keratinocyte growth factor (KGF) receptors ligand; The DPP IV inhibitor; REG protein receptor part; The growth hormone receptor part; Prolactin (PRL) receptors ligand; IGF (IGF) receptors ligand; PTH GAP-associated protein GAP (PTHrP) receptors ligand; HGF (HGF) receptors ligand; Bone morphogenetic protein (BMP) receptors ligand, transforming growth factor β (TGF-β) receptors ligand; The laminin receptor part; Vasoactive intestinal peptide (VIP) receptors ligand; Fibroblast growth factor (FGF) receptors ligand; The keratinocyte growth factor receptors ligand; Nerve growth factor (NGF) receptors ligand; Pancreas islet regeneration GAP-associated protein GAP (INGAP) receptors ligand; The activin A receptors ligand; VEGF (VEGF) receptors ligand; Erythropoietin(EPO) (EPO) receptors ligand; Pituitary adenylate cyclase activated polypeptides (PACAP) receptors ligand; Granulocyte colony stimulating factor (G-CSF) receptors ligand; Granulocyte macrophage colony stimulating factor (GM-CSF); Platelet derived growth factor (PDGF) receptors ligand and secretin-receptor ligands.

As used herein, term " diabetes " meaning be insulin short-lived, produce AIA or the excessive any physiology indication of blood sugar. Diabetes illustrate, but are not limited to type i diabetes, type ii diabetes, gestational diabetes mellitus and prediabetes state. As used herein, term " mammal " has any member's of mammal common implication, comprises the people.

In relevant embodiment, FACGINT is glucagon-like peptide 1 (GLP-1) receptors ligand; Glucagon-like peptide 2 (GLP-2) receptors ligand; Perhaps growth hormone receptor ligand family member can be growth hormone, such as human growth hormone (HGH) (HGH), galactagogin (PL) or prolactin (PRL) or exendin, such as exendin-4.

Another characteristic of the present invention provides the method for the treatment of diabetes, and the method comprises a large amount of cells and at least a composition in having FACGINT and gastrin/cck receptor part exsomatized and contact that needing only FACGINT is not the EGF receptors ligand; This cell is applied to the mammal that need to use, thereby the treatment diabetes. In an embodiment of the method, cell is from body, that is, and and from the experimenter. Alternatively, cell is from another individuality in the same species, perhaps even from another species. In the method for use cell that exsomatizes, cell can be the pancreatic duct cell. Pancreatic cell is the source of pancreas islet precursor. Another embodiment of this method comprises, before transplanting step, with said composition ex vivo treatment cell. Relevant method further comprises, before the contact procedure, and this cell of cultured in vitro.

Usually, term " administration " or " contact " meaning is the composition that the effective dose of insulin secretory cell amount in the raising mammal is provided to the user of method.

Usually in these methods, composition is the whole body administration. In addition, in the composition amount of FACGINT basically less than the least effective dose (LED) that when lacking gastrin/cck receptor part, reduces the required FACGINT of blood-glucose in the diabetes mammal. The method can comprise further that measurement is selected from lower group parameter: the β cell mass that blood-glucose, serum glucose, the glycosylated hemoglobin of blood, pancreatic beta cell group, serum insulin, pancreas insulin content and form tolerance are measured. Generally speaking, the technical staff in diabetes diagnosis field will measure blood or Glucose in serum level after fasting, that is, during experimenter or experimenter do not take food, the typical duration of this diagnosis. Canonical measure is to measure fasting blood glucose, or FBG.

Method can comprise further that here measurement is selected from lower group parameter in the body: the amount of insulin secretory cell, the glucose response of insulin secretory cell, the amount of pancreas islet precursor propagation and the amount of mature insulin secretory cell, these measurements are carried out in mammal, and wherein this mammal is animal used as test such as KKAy Mouse (NOD mouse) or is induced (the rodent of using Streptozotocin (streptozoticin) or STZ) suffering from diabetes.

Here another embodiment is the method for inducing the regeneration of pancreas pancreas islet in mammal, the composition that wherein comprises FACGINT and gastrin/cck receptor ligand combination to administration, as long as FACGINT is not the EGF receptors ligand, be enough to improve the amount of pancreas islet precursor propagation in the pancreatic tissue, therefore induce the regeneration of pancreas pancreas islet.

Here another embodiment is the method for inducing pancreas pancreas islet regeneration in the mammal, the method comprises the composition that comprises FACGINT and gastrin/cck receptor ligand combination, wherein FACGINT is not the EGF receptors ligand, is enough to improve the amount of pancreas islet secretion β cell quantity in the mammal.

Therefore, embodiment of the present invention are the compositions that comprise gastrin/cck receptor part and FACGINT, as long as FACGINT is not the EGF receptors ligand. The composition of inducing pancreas islet precursor propagation to form the effective dose of ripe insulin secretory cell quantity increase is provided in some embodiments. Similarly, provide in some embodiments the composition of inducing the pancreas islet precursor to be differentiated to form the effective dose of mature insulin secretory cell.

The composition that contains pharmaceutical acceptable carrier can be provided.

The kit for the treatment of diabetes also is provided here, contains the composition, container and the operation instruction that comprise gastrin/cck receptor part and FACGINT, as long as FACGINT is not the EGF receptors ligand. The a small amount of composition that contains one or more UDs can be provided. The composition of kit further comprises pharmaceutically acceptable carrier.

The present invention's another characteristic here is the method for amplification and differentiated stem cells formation insulin secretory cell in implanting the diabetes acceptor of cell, it is included in and implants stem cell in the acceptor, and use each composition among the gastrin that contains effective dose/cck receptor part and the FACGINT to acceptor, as long as FACGINT is not the EGF receptors ligand. In this method, for example can from people or pig, obtain cell. In addition, from pancreas islet, umbilical cord, embryo or stem cell line, obtain the cell of implantation. Alternatively, the method for amplification and differentiated stem cells formation insulin secretory cell is in the diabetes acceptor of cell: implant cell in acceptor; And comprise each sustained-release composition in the gastrin of effective dose/cck receptor part and FACGINT or the EGF receptors ligand.

Generally speaking according to these methods, gastrin/cck receptor part is gastrin. In relevant embodiment, gastrin is Gastrin-17, and for example, gastrin is people's gastrin 1-17Leu15.

In addition, according to these methods, implanting cell in acceptor is the approach that uses as be injected directly into organ and intravenous administration. Injection cell part is transported in the organ, for example, and pancreas, kidney and liver. In addition, the injection cell is through skin or wear liver and be transported to portal vein. In these methods any, conduit can be inserted into from or lead in the vein or artery of organ, use during operation imaging technique, such as ultrasonic or MRI.

The local cell of carrying can be selected from several technology, comprises endoscopic retrograde choledochopancreatography (ERCP); The EUS guided fine needle is carried (EUS-FNAD); Be expelled in the arteria pancreatica; Be expelled in the portal vein; Injection in the pancreas; And be expelled in the arteria hepatica. In these technology, the user can be by a kind of guiding in several imaging techniques that comprise ultrasonic or MRI during operation.

Another characteristic provided by the invention is to reduce the method for the required transplanting for the treatment of people diabetes stem cell amount, the method comprises to acceptor uses among the gastrin of effective dose/cck receptor part and the FACGINT each, implanting cell when giving other same receptor effective dose in shortage compares, the amount of cell reduces, as long as FACGINT is not the EGF receptors ligand.

In related embodiment of the present invention, invention provides the composition that comprises gastrin/cck receptor part and at least a FACGINT, as long as FACGINT is not the EGF receptors ligand. Provide and contain the composition of effectively inducing the pancreas islet precursor to be differentiated to form mature insulin secretory cell dosage. The composition that contains pharmaceutical acceptable carrier can further be provided.

The kit for the treatment of diabetes is provided, and this kit contains the composition that comprises at least a FACGINT, as long as FACGINT is not the EGF receptors ligand, and gastrin/cck receptor part, container and operation instruction.

Use the reagent that Immunosuppression reacts any can further the comprising to the experimenter in the said method. In any in said method, the component of combination can be carried simultaneously, for example, in 1 hour or in one day, perhaps can carry in proper order, for example, with surpass one day, super two days later or more days, surpass the interval in a week.

The exemplary reagent of Immunosuppression reaction is medicine, for example, is at least a in lower group: rapamycin; Corticosteroid; Imuran; Mycophenolate; Cyclosporin; Endoxan; Amethopterin; Ismipur; FK506; DSG; FTY 720; Mitoxantrone; The 2-amino-1,3-propanediol; 2-amino-2[2-(4-octyl phenyl) ethyl] propane-1,3-glycol hydrochloric acid; 6-(3-dimethyl-amino propionyl) Forskolin; And demethimmunomycin.

Alternatively, the exemplary reagent of Immunosuppression reaction is protein, and for example, this albumen comprises the amino acid sequence of antibody. Therefore, the reagent of Immunosuppression reaction is at least a in following: hul 124; BTI-322; Allotrap-HLA-B270; OKT4A; Enlimomab; ABX-CBL; OKT3; ATGAM; Basiliximab; Daclizumab; Thymoglobuline (thymoglobulin); ISAtx247; Medi-500; Medi-507; Alefacept; The Yi Lizuo monoclonal antibody; Infliximab; And interferon. Give sequentially reagent or the simultaneously administration of the reaction of pancreas islet regenerative therapy composition and Immunosuppression.

In certain embodiments, whole body gives at least a in the reagent of pancreas islet regenerative therapy composition and Immunosuppression reaction. For example, pancreas islet regenerative therapy composition is used as pill. Therefore, by in intravenous, subcutaneous, the peritonaeum or the intramuscular approach give at least a in the reagent of pancreas islet regenerative therapy composition and Immunosuppression reaction. In addition, in certain embodiments, the oral reagent that gives the Immunosuppression reaction. Usually, the reagent of Immunosuppression reaction is at least a in FK506, rapamycin and the daclizumab. In addition, according to any embodiment that is different from here in those some experiment parameter methods of requirement measurement, the experimenter is the people.

The kit that is used for the treatment of diabetic subjects here also is a kind of characteristic, and the INT composition that kit comprises container, immunodepressant and comprises FACGINT is not as long as FACGINT is the EGF receptors ligand.

I.N.T. also is provided here^TMThe pharmaceutical composition of the sustained release of therapeutic combination, said composition comprises: the gastrin-receptor part; With EGF receptors ligand or FACGINT; Wherein at least a among gastrin-receptor part or EGF receptors ligand or the FACGINT is extended release preparation.

This composition can further comprise immunodepressant. The exemplary embodiment of extended release preparation is the Pegylation of at least a composition component, and at least a component is in based on the preparation in the liposome of many bubbles lipid. The example of EGF receptors ligand is to be selected from EGF and TGF α. The example of FACGINT is to be selected from least a in lower group: the glucogan-like peptide 1 receptor part; The glucagon-like peptide 2 receptors ligand; Intestines gastric inhibitory polypepitde (GIP) receptors ligand; Keratinocyte growth factor (KGF) receptors ligand; Inhibitors of dipeptidyl IV; REG protein receptor part; The growth hormone receptor part; Prolactin (PRL) receptors ligand; IGF (IGF) receptors ligand; PTH GAP-associated protein GAP (PTHrP) receptors ligand; HGF (HGF) receptors ligand; Bone morphogenetic protein (BMP) receptors ligand, transforming growth factor β (TGF-β) receptors ligand; The laminin receptor part; Vasoactive intestinal peptide (VIP) receptors ligand; Fibroblast growth factor (FGF) receptors ligand; The keratinocyte growth factor receptors ligand; Nerve growth factor (NGF) receptors ligand; Pancreas islet regeneration GAP-associated protein GAP (INGAP) receptors ligand; The activin A receptors ligand; VEGF (VEGF) receptors ligand; Erythropoietin(EPO) (EPO) receptors ligand; Pituitary adenylate cyclase activated polypeptides (PACAP) receptors ligand; Granulocyte colony stimulating factor (G-CSF) receptors ligand; Granulocyte macrophage colony stimulating factor (GM-CSF); Platelet derived growth factor (PDGF) receptors ligand; And secretin-receptor ligands. Alternatively, the EGF receptors ligand is low-molecular-weight drug. Composition can be formulated into for parenteral. Alternatively, composition can be formulated into for oral administration.

That composition can be formulated into is subcutaneous be used to being selected from, in the peritonaeum, intravenous and intramuscular injection administration. In one embodiment, at least a being formulated into for the whole body administration among gastrin-receptor part or EGF receptors ligand or the FACGINT.

In addition, above-mentioned composition can be formulated into for the part and carry, and for example, carry the part of targeted pancreatic. Local exemplary type of carrying comprises endoscopic retrograde choledochopancreatography (ERCP); The EUS guided fine needle attracts administration (EUS-FNAD); Be expelled in the arteria pancreatica; Be expelled in the portal vein; Injection in the pancreas; And be expelled in the arteria hepatica. During operation, these can carry out in conjunction with imaging technique such as ultrasonic or MRI.

In some embodiments, composition is formulated into be used to the method for administration that is selected from through skin and mucous membrane conveying. Any being mixed with for carrying by mechanical device in these compositions, the associating extended release preparation perhaps alternatively uses mechanical device to carry preparation in the time that continues. Device is, for example, and degradable implant; Transdermal patch; Conduit; Implantable pump; Through the skin pump; Infusion pump; And Iontophoretic device.

Above-mentioned composition and pharmaceutical composition can be mixed with and be suitable for being selected from following administration: in subcutaneous, the peritonaeum, in intravenous and the pancreas. For example, intravenous route is to be injected into portal vein. But operative installations, this device can be pumps. With extended release preparation with in the above-mentioned method and composition some, but topical. For example, can carry and topical by the approach that is selected from lower group: endoscopic retrograde choledochopancreatography (ERCP); The EUS guided fine needle attracts administration (EUS-FNAD); Be expelled in the arteria pancreatica; Be expelled in the portal vein; Injection in the pancreas; And be expelled in the arteria hepatica. Alternatively use extended release preparation and device, but the whole body administration. Can provide and contain the effective dose of medicine compositions. Here the kit that comprises at least a dosage of any composition in the above-mentioned extended release preparation also is a kind of characteristic.

Reduce and use I.N.T.^TMThe method of the frequency of combination treatment diabetic subjects also is a kind of characteristic, and the method comprises that the component with at least a composition is prepared into extended release preparation; And use composition according to the scheme that the time interval between the treatment is longer than so preparation and the identical composition of other side to the experimenter. Can carry administration by being selected from following approach: endoscopic retrograde choledochopancreatography (ERCP); The EUS guided fine needle attracts administration (EUS-FNAD); Be expelled in the arteria pancreatica; Be expelled in the portal vein; Injection in the pancreas; Perhaps be expelled in the arteria hepatica. Can use the imaging technique catheter guidance to be put into suitable position in these operating periods.

Improve I.N.T. in the diabetic subjects^TMThe method of composition effect also is a kind of characteristic, and the method comprises: use the component with at least a composition to the experimenter and be formulated into the I.N.T. that produces sustained release^TMComposition; Relatively give said composition amount treatment experimenter's effect and do not have the component of such preparation and the effect of the identical composition of other side, the amount that reduces or eliminates the required sustained release reagent preparation of experimenter's diabetic symptom by mensuration reduces, so that have the I.N.T. of sustained release compositions formulated^TMThe effect of composition improves. In an embodiment of this method, relatively effect is the toxicity of further analysed composition so that with the component that does not have such preparation and the identical I.N.T. of other side^TMThe toxicity of composition is compared, and gives less or slighter harmful symptom behind the composition and shows that having the toxicity that at least a component is formulated the composition that produces sustained release reduces. In any in these methods, the exemplary extended release preparation of component is Pegylation and based on the liposome of many bubbles lipid.

Improve I.N.T. at these^TMIn composition effect method any, the component with extended release preparation is the EGF receptors ligand that is selected from EGF and TGF α. Alternatively, in the embodiment of these methods, FACGINT is selected from least a in lower group: the glucogan-like peptide 1 receptor part; The glucagon-like peptide 2 receptors ligand; Intestines gastric inhibitory polypepitde (GIP) acceptor; Keratinocyte growth factor (KGF) receptors ligand; Inhibitors of dipeptidyl IV; REG protein receptor part; The growth hormone receptor part; Prolactin (PRL) receptors ligand; IGF (IGF) receptors ligand; PTH GAP-associated protein GAP (PTHrP) receptors ligand; HGF (HGF) receptors ligand; Bone morphogenetic protein (BMP) receptors ligand, transforming growth factor β (TGF-β) receptors ligand; The laminin receptor part; Vasoactive intestinal peptide (VIP) receptors ligand; Fibroblast growth factor (FGF) receptors ligand; The keratinocyte growth factor receptors ligand; Nerve growth factor (NGF) receptors ligand; Pancreas islet regeneration GAP-associated protein GAP (INGAP) receptors ligand; The activin A receptors ligand; VEGF (VEGF) receptors ligand; Erythropoietin(EPO) (EPO) receptors ligand; Pituitary adenylate cyclase activated polypeptides (PACAP) receptors ligand; Granulocyte colony stimulating factor (G-CSF) receptors ligand; Granulocyte macrophage colony stimulating factor (GM-CSF); Platelet derived growth factor (PDGF) receptors ligand; And secretin-receptor ligands. Even being further used as alternative, component is low-molecular-weight drug. In these any methods, administration is to carry by the approach that is selected from lower group: parenteral, oral, in skin, subcutaneous, mucous membrane, peritonaeum, in the intravenous, pancreas and intramuscular.

For example, administration produces local distribution. In addition, the method comprises the composition that gives effective dose. Even further, the method comprises, before administration, composition is mixed with is suitable for using sustained release device. For example, this device is selected from: degradable implant; Transdermal patch; Conduit; Implantable pump; Through the skin pump; Infusion pump; And Iontophoretic device. For example, this device is pump. In any in these methods, administration can be to be expelled in the portal vein by intravenous route.

The method that reduces the frequency for the treatment of diabetic subjects also is a kind of characteristic, and the method comprises that preparation discharges to the experimenter by long-time continuous and uses I.N.T.^TMThe device of composition; To experimenter's generator; And repeatedly treat the experimenter by replacing or refilling device, for example, device is pump. Pump can be through the skin pump; Carbon fluorine (flurorcarbon) propellant pump; Osmotic pumps; Miniosmotic pump; Implantable pump; Perhaps infusion pump. Alternatively, device is selected from lower group: degradable implant; The implant of non-degraded; The mucosal adhesive implant; Transdermal patch; Conduit; And Iontophoretic device. The implant of exemplary non-degraded is silicon rubber. Other example of degradable implant can be to be selected from least a in the lower group of material: starch; Vinyl starch; DPG ester diacrylate (DPGDA); Tripropylene glycol ester diacrylate (TPGDA); Pectin; Acetylcellulose; Cellulose propionate; Cellulose acetate-butyrate; Cellulose-acetate propionate (CAP); Hydroxypropyl cellulose (HPC); Hydroxypropyl cellulose/cellulose-acetate propionate (HPC/CAP); Methyl methacrylate (MMA); Butyl methacrylate (BMA); Methylol methyl acrylate (HEMA); Acetic acid Octyl Nitrite (EHA); Octadecyl methacrylate (ODMA); And ethylene glycol dimethacrylate (EGDMA).

The method of amplification and differentiated stem cells formation insulin secretory cell also is a kind of characteristic in the diabetes acceptor of cell, has step: implant cell in acceptor; With the gastrin that comprises effective dose/cck receptor part; And each the sustained-release composition in FACGINT or the EGF receptors ligand, wherein amplification and differentiated stem cells formation insulin secretory cell in acceptor.

The composition that comprises glucagon-like-peptide-1 (GLP-1) receptors ligand and gastrin/cck receptor part for the treatment of diabetes also is a kind of characteristic. For example, the GLP-1 receptors ligand is GLP-1. The composition that comprises growth hormone (GH) receptors ligand and gastrin/CCK receptors ligand for the treatment of diabetes also is a kind of characteristic. For example, GH is human growth hormone (HGH) (HGH). The composition that comprises prolactin (PL) receptors ligand and gastrin/cck receptor part for the treatment of diabetes also is a kind of characteristic. For example, PL is people PL. In any one of these compositions, exemplary gastrin is that the 15th amino acids is 17 the amino acid whose gastrin I that have of leucine residue. Any in these compositions can further have immunodepressant. Any in these compositions further can be mixed with for sustained release.

The method of using the treatment diabetic subjects that comprises gastrin/cck receptor part and glucagon-like-peptide-1 (GLP-1) receptors ligand composition to the experimenter also is a kind of characteristic. The method of using the treatment diabetic subjects of the composition that comprises gastrin/cck receptor part and growth hormone (GH) receptors ligand to the experimenter also is a kind of characteristic. The method of using the treatment diabetic subjects of the composition that comprises gastrin/cck receptor part and prolactin (PL) receptors ligand to the experimenter also is a kind of characteristic. Give immunodepressant any the comprising in these methods. Any in these methods can further comprise and use sustained release device to give at least a in receptors ligand or the reagent. Any can further comprising at least a sustained release that is mixed with in receptors ligand or the reagent in these methods. Any diabetic subjects that can be used for suffering from type i diabetes or type ii diabetes in these methods.

The method that enlarges the functional β cell mass of Islet allografts in the diabetic subjects acceptor of the graft of purifying pancreas islet also is a kind of characteristic, and the method gives gastrin cck receptor part and the FACGINT of mammal effective dose.

Comprise the pancreas islet goods are transplanted in the diabetic subjects; And the method that gives the treatment people diabetes of the gastrin of experimenter's effective dose/cck receptor part and FACGINT also is a kind of characteristic. Therefore in this method, FACGINT comprises that it is the hprl receptor part of prolactin.

The detailed description of specific embodiments

Composition (the I.N.T. of pancreas islet regenerative therapy^TM) comprise gastrin/cck receptor part, and EGF receptors ligand or the additional gastrin pancreas islet regenerative therapy factor (FACGINT).

By using FACGINT and gastrin/cck receptor part, for example, the combination of gastrin, the treatment of diabetes have produced the increase of the effectiveness, effect and the effectiveness that surpass separately the wondrous level for the treatment of with any one-component. Owing to this reason, the term FACGINT meaning used herein " is replenished " the gastrin administration. Term " replenishes " synergy that might not refer between implicit gastrin and the FACGINT, and this term meaning is to compare with FACGINT with using gastrin on the contrary, and is more effective for correcting diabetes with the administering drug combinations of used dosage in the combination.

Any composition or the compound that will mean combination, interaction with receptor related term " receptors ligand " particular ligand or stimulate this receptor used herein.

Term " FACGINT " comprises one or more reagent in a large amount of various growth factors and growth hormone, the modifying factor hormone, part and the effector of related one or more acceptors in the combination of these growth hormone and growth factor, usually the implication of understanding with these terms is identical, for example but be not limited to: PTH GAP-associated protein GAP (PTHrP) receptors ligand such as PTHrP (PTHrP; Garcia-Ocana, A. etc., 2001, J.clin.Endocrin.Metab. 86:984-988); HGF (HGF) receptors ligand such as HGF (HGF; Nielsen, J. etc., 1999, J Mol Med 77:62-66); Fibroblast growth factor (FGF) is such as FGF, keratinocyte growth factor (KGF) receptors ligand such as KGF; Nerve growth factor (NGF) receptors ligand such as NGF; Intestines gastric inhibitory polypepitde (GIP) acceptor such as GIP; Transforming growth factor β (TGF β) receptors ligand such as TGF β (disclosed U.S. Patent application 2002/0072115 on June 13rd, 2002), laminin receptor part such as EHS-laminin; Pancreas islet regeneration GAP-associated protein GAP (INGAP) receptors ligand such as INGAP; Bone morphogenetic factor (BMP) receptors ligand such as BMP-2; Vasoactive intestinal peptide (VIP) receptors ligand such as VIP; Glucagon-like peptide-1 receptor part such as GLP-1 and exendin-4, glucagon-like peptide 2 (GLP-2) receptors ligand such as GLP-2, and by suppressing related enzyme fully, inhibitors of dipeptidyl IV (the Hughes of remote-effects GLP-1 level, T. etc., 2002, Am Diabetes Assoc Abstract 272-or); REG receptors ligand such as REG albumen; Growth hormone (GH) receptors ligand such as GH, prolactin (PRL) receptors ligand such as PRL and galactagogin (PL); IGF (1 type and 2 types) receptors ligand such as IGF1 and IGF-2; Erythropoietin(EPO) (EPO) receptors ligand such as EPO (http: ∥ www.drinet. org/html/august_2002._htm); β cytokine (betacellulin) (member who also is considered to EGF family); Activin A receptors ligand such as activin-A; VEGF (VEGF) receptors ligand such as VEGF; Bone morphogenetic factor (BMP) receptors ligand such as BMP-2; Vasoactive intestinal peptide (VIP) receptors ligand such as VIP; VEGF (VEGF) receptors ligand such as VEGF; Pituitary adenylate cyclase activated polypeptides (PACAP) receptors ligand such as PACAP; Granulocyte colony stimulating factor (G-CSF) receptors ligand such as G-CSF; Granulocyte macrophage colony stimulating factor (GM-CSF) receptors ligand such as GM-CSH; Platelet derived growth factor (PDGF) receptors ligand such as PDGF and secretin-receptor ligands such as secretin.

Any in growth factor, enzyme, enzyme inhibitor, peptide, albumen and the hormonal compounds of exemplary FACGINT for being represented as here, all known analogs, variant and derivative, natural generation or all should be considered to be equal to FACGINT by mutagenesis or design and synthetic preparation. Conjugate also is considered in equivalent, that is, and and the composition of deriving by adding one or more chemical groups, and composition thereof. Can change encoding gene, for example, produce the FACGINT analog by oligonucleotides mutagenesis, such as people's analog of recombinating. In addition, can change one or more than an amino acid residue itself or position by directed mutagenesis. Can increase by conjugate the one-level amino acid sequence of albumen, as by glycosylation, acidylate or by adding any other additional molecule, such as one or more lipids, phosphate and/or acetyl group. In addition, can be by the single amino acids residue in oxidation, reduction or other derivatization modification chain. Can cut any fragment of FACGINT acquisition retentive activity. The activator of FACGINT, pro-drug or metabolite are equal to FACGINT. Can be with whole polypeptide or albumen or any fragment and any other peptide or protein fusion, such as immunoglobulin (Ig) and other cell factor. The variant that is in nature the FACGINTs of protein can be produced by alternative splicing, proteolysis cutting or other posttranslational modification of primary transcribe, comprises dimerization, polymerization, phosphorylation, glycosylation, sulphation and desamidation. Conjugate can comprise, for example, comprises the composition of the FACGINT on the polymer that is coupled to the non-natural generation, and wherein polymer comprises polyalkylene glycol moiety. This term also comprises the derivative of modifying one or more amino acid residues acquisitions of parental generation peptide by chemical method, for example by alkylation, acidylate, formation ester or formation acid amides. In addition, induce FACGINT reagent synthetic or simulation FACGINT effect to be considered to be equal to compound. Any or multiple compounds among the exemplary FACGINTs shown in " FACGINT " of singulative can refer to here.

Here in several usages of term, elaborated term FACGINT and do not comprised the EGF receptors ligand, also very clear in context equally. Yet EGF receptors ligand such as EGF and TGF can replenish gastrin and be used for correct diabetic disease states, and therefore are defined exemplary FACGINTs here, are included in as component in other embodiment of here composition, method and kit. Described in the past and comprised that associating gastrin/cck receptor part gave certain embodiment (U.S. Patent number 5 of the I.N.T. treatment of EFG receptors ligand, 885,956 and 6,288,301), these lists of references do not use these reagent in as shown here some combination and the preparation.

Term used herein " pancreatic progenitor cell " or " β cell ancestors " or " pancreas islet precursor " are the precursors that can be differentiated to form pancreas β islet cells, its may have or may not have a stem cell can be with the characteristic of unlimited mode self-replacation. " β cytothesis " or " pancreas islet regeneration " meaning is by being differentiated to form new β cell, its may have or may not have a stem cell can be with the characteristic of unlimited mode self-replacation. Looking like with " glucose response " mode excreting insulin is according to the concentration of glucose excreting insulin in the blood. In the normal mammal of physiological function, along with the raising of blood glucose levels, the β emiocytosis insulin of pancreas islet, that is, generation is replied and secretion inducing insulin to blood glucose levels.

Acceptor " activator " is hard-core any composition, for example, endogenous is found in mammal PGF or cell factor or variant or its part, perhaps analog, perhaps any plan peptide (peptidomimetic) or low-molecular-weight drug, it has the ability of acceptor of the polypeptide factor of combination and activation of endogenous discovery. Any in growth factor described herein or the cell factor, equivalent is considered to be in essentially identical material on the amino acid sequence, for example, with naturally occurring peptide described herein or albumen have 50% sequence identical, have 60% sequence identical, have 70% sequence identical or have 80% sequence identical. In other embodiments, the activator composition comprises the agonist induction agent here, it is envisioned as when giving animal or offer cell in the culture, organ or tissue, can increase the material of the amount of the activator that animal, cell, organ or tissue produce. For example, Prolactin-Releasing Peptide stimulates the secretion of prolactin. Receptors ligand comprises receptor stimulating agent in the scope of definition, to the acceptor of any specific FACGINT, no matter whether activator is structurally relevant with FACGINT.

The present invention provides in one embodiment by comprising gastrin/cck receptor part, for example gastrin, and FACGINT, for example both combination treatment diabetic disease states of GLP-1, PRL or GH such as the method for diabetes. Do not limited by any special mechanism, being enough to affect the amount that the pancreas islet precursor is divided into the mature insulin secretory cell with every kind provides gastrin/cck receptor part and FACGINT. Each of FACGINT and gastrin/cck receptor part can whole body or topical in the composition. Alternatively, a kind of in FACGINT and gastrin/cck receptor part or both, the cell in-situ that can be provided the expression vector that contains the nucleic acid fusion constructs is expressed. Fusion constructs typically comprises preprogastrin (preprogastrin) peptide precursor coded sequence, and the coded sequence that also comprises FACGINT.

Give gastrin/cck receptor part and EGF receptors ligand and can reach the effect that prolongs pancreatic cell regeneration, like this benefit for the treatment of remain to treat stop after for a long time. See disclosed PCT application on July 18th, 2002 PCT/US02/00685 (WO 02/055152). Lasting longer than the duration that gives the composition scheme for the treatment of benefit.

With the treatment diabetes that provide, particularly composition and the method for the diabetes of juvenile morbidity, and give this combination of the factor or be provided for the whole body administration by therapeutic, or be used for the composition that original position is expressed in pancreas, obtain the cytothesis of residual multipotency pancreatic duct and be differentiated to form ripe insulin secretory cell.

It is to stimulate β cytothesis that elimination needs the method for the β cell exchange of cell transplantation. Have limited power of regeneration although research and propose in early days the β cell, now recognized gradually that the insulin secretion β cell of pancreas comprises the dynamic cellular group. The β cell mass can enlarge by the propagation (β cellular replication) of existing β cell. At pregnancy duration, prolactin, growth hormone (Holstad, M. etc., J.Endocrinol.163:229-234) and galactagogin (Nielsen, J.H., etc., J.Mol.Med.77:62-66,1999) stimulate the propagation of β cell to increase cell mass. Yet this expansion of cell mass relies on the hormonal stimulation that continues. Divide the puerperium, the cell mass that causes enlarging is replied in the reduction generation of prolactin and galactagogin be reduced to non-pregnant level (Logothetopoulos, J., (Logothetopoulos, J. (1972) in Handbook of Physiology (Am.Physiol.Soc., Washington, DC), Section 7, and Chapter 3, pp67-76).

Know that from this physiologic information to produce an importance of replying be the β cell mass that enlarges sustainable effective time after the treatment with growth factor stops whether to giving gastrin-receptor part and EGF receptors ligand or FACGINT estimating β cytothesis. Use extended release preparation, associating I.N.T. composition, associating or combined immunization inhibitor not can be avoided frequently going to medical institutions, needing perhaps to avoid self-drug therapy.

Regeneration according to the β cell is measured in the increase of cell quantity and cell mass causes the increase of plasma insulin level or pancreas insulin content. The effect prolongation is the expected result of I.N.T. in the treatment of diabetes.

One embodiment of the invention provide uses FACGINT together with the method and composition of the improvement of gastrin/cck receptor ligands for treating diabetes. The present invention provides the combination of gastrin and FACGINT to use separately higher effect, usefulness and the effectiveness that component was obtained to obtain ratio in one embodiment, and the treatment that causes making up is than improving. Make the minimizing of blood-glucose be higher than separately minimizing with the blood-glucose of observing after the component treatment with the treatment of gastrin and FACGINT combination, the long-time continuous decrease of blood-glucose after stopping to treat. Phrase used herein " FACGINT " also can mean " a kind of and multiple FACGINTs " or " at least a FACGINT ".

In suffering from the part pancreas excision model of slight diabetes, β cell stimulatory agents Exendin (GLP-1 analog) improves glucose-tolerant and increases β cell mass (Xu, G. etc., Diabetes 48:2270-2276,1999). Yet, do not draw the glucose-tolerant of raising and the causality between the β cytothesis with having concluding. GLP-1 is a kind of exemplary FACGINT as shown here, appetite-suppressing and the clearance rate that improves glucose by reducing insulin resistance when giving separately, a kind of may with the irrelevant process of β cytositimulation. Therefore, it is not the result of β cytositimulation that the raising of viewed glucose-tolerant is pointed out in the discovery that plasma insulin level is lower rather than higher in the Exendin treatment group. In addition, impact is complicated on the β Growth of Cells to estimate Exendin by the pancreas excision model of the diabetes used in these researchs. Because the inflammation due to the surgical excision of pancreas causes acting on the expression of the growth factor of pancreas islet, such as gastrin and TGF α, itself stimulates pancreas islet regeneration by their. In fact, reported that β cytothesis increases (Bonner-Weir, S., Diabetes 42:1715-1720,1993 after the simple pancreatectomy; Sharma, A., etc., Diabetes 48:507-513,1999). Therefore, in the past and do not know that Exendin can stimulate β cytothesis to increase when lacking the growth factor that these pancreatectomies induce.

Term used herein " diabetes " meaning is any diabetic symptom that manifests in any mammal, comprise experimental animal model, and all kinds that comprise the people, reduce or blood glucose levels is slight raises as the prediabetes state of characteristics such as I type and II type diabetes, early diabetes and so that insulin is slight. " prediabetes state " described and suspected the mammal with diabetes or relevant disease, for example, formally be not diagnosed as diabetes, but showing symptom aspect insulin or the glucose level, because the obesity in family history, heredodiathesis or the type ii diabetes or suffered from the past diabetes or relevant disease and had risk of recurrence to cause easily suffering from diabetes or relevant disease.

As used herein, term " gastrin/cck receptor part " comprises any combination, interaction or stimulates the compound of gastrin/cck receptor. The United States Patent (USP) 6 of delivering in September 11 calendar year 2001,288, provided the example of this gastrin/cck receptor part in 301, the various forms that comprises gastrin is such as gastrin 34 (big gastrin), G17 (little gastrin) and gastrin 8 (small gastrin); Various forms of cholecystokinins such as CCK 58, CCK 33, CCK 22, CCK 12 and CCK 8; And other gastrin/cck receptor part. Generally speaking, the total c-terminus amino acid sequence Trp-Met-Asp-Phe-acid amides of gastrin/cck receptor part. What also be considered is active analogue thereof, fragment and other modification of above-mentioned substance, the partial agonist that comprises peptide and non-peptide agonists or gastrin/cck receptor, such as A71378 (Lin etc., Am.J.Physiol.258 (4Pt1): G648,1990).

Small-sized gastrin such as G17 are that synthetic peptide is commercially available by the synthetic economically preparation of peptide. Synthetic people's G17 is that methionine or leucic people's G17 also can be from Bachem AG such as the 15th amino acids, Bubendorf, and Switzerland, and obtain from Researchplus. Gastrin/cck receptor part also comprises active analogue thereof, fragment and other modification of above-mentioned part, it for example has common amino acid sequence with endogenous mammal gastrin, and it is identical or 70% identical or 80% identical for example to have 60% sequence. Such part also comprises to be increased from organizing stores location to secrete the compound of endogenous gastrin, cholecystokinin or similar active peptide. The example of these compounds is Omeprazoles (omeprazole) of gastrin releasing peptide, gastric acid secretion inhibiting and strengthens the soybean trypsin inhibitor that CCK stimulates.

In the individuality that needs are used, the method for the treatment of diabetes comprises and gives the composition that individuality provides gastrin/cck receptor part and FACGINT. Do not limited by any special mechanism, provide gastrin/cck receptor part and FACGINT to be enough to affect the dosage that the pancreas islet precursor is divided into the mature insulin secretory cell.

Term used herein " treatment " or " improvement " meaning are the one or more symptoms that reduce or eliminate diabetes. The method of the treatment diabetes that provide here comprises and gives the diabetes mammal, not limited by any special mechanism, the gastrin of differentiation and regeneration amount/cck receptor part and FACGINT stimulates pancreas islet to regenerate to increase the quantity of function glucose reactivity insulin excreting beta cell in the pancreas. The method is to diabetes, generally includes the I type or the juvenile form diabetes are effective. The combination of gastrin and FACGINT will cause the pancreas islet regenerative response to significantly improve, and surpass with the viewed pancreas islet regenerative response of one-component. Exemplary gastrin/cck receptor part is gastrin, and exemplary FACGINTs is GLP-1, PRL and GH.

Here another embodiment of the invention is to comprise the mammiferous pancreatic tissue of transplanting with gastrin/cck receptor part and FACGINT ex vivo treatment, and treated pancreatic tissue is input to method in the mammal. In this method, exemplary gastrin/cck receptor part is gastrin again, and exemplary FACGINTs is GLP-1, PRL and GH.

In another embodiment, the invention provides the method for gastrin/cck receptor ligand stimulation, the method comprises to pancreatic cell to be provided chimeric insulin promoter-gastrin fusion and expresses this gene. In another embodiment, the method that provides FACGINT to stimulate comprises expressing importing mammiferous FACGINT gene by transgenosis, for example, and the gene of coding FACGINT, for example, GLP-1, PRL or GH. Can provide gastrin/cck receptor ligand gene by transgenosis similarly, preferably as at U.S. Patent number 5,885, the people's preprogastrin peptide precursor gene shown in 956.

Should unrestrictedly comprise mammiferous any member such as term mammal used herein, such as people, ape, rodent such as mouse or rat, dog, cat, very important animal or protein porcine, goat, sheep, horse or ape such as gorilla or chimpanzee on the agricultural. According to explanation here, single mammal may be non-diabetic, prediabetes or suffer from diabetes.

The mode of whole body administration includes, but not limited in skin, sheath, in the intramuscular, peritonaeum, in the intravenous, subcutaneous, nose and oral route. Can give compound by any easily approach, for example, by transfusion or single injection, by through epithelium or mucocutaneous lining (for example, oral mucosa, mucous membrane of rectum, vagina mucosa, schneiderian membrane and intestinal mucosa, Deng) absorption, and can be with other bioactive agents administration. The exemplary approach of administration is the whole body administration, for example by hypodermic injection.

Term used herein " prolactin " meaning is known in the rho factor field such as this term, have any polypeptide of basic sequence similitude with the mammiferous lactagogue of endogenous, for example, people's prolactin, it has the activity of prolactin. Endogenous people prolactin is 199 amino acid whose polypeptide that pituitary gland produces. This term comprises the prolactin analog, and it is disappearance, the insertion of endogenous prolactin or replaces mutant, and retentive activity, comprises the prolactin of other species and the variant of natural generation. The function of prolactin comprises as at U.S. Patent number 6,333,031 (activated amino acid sequence) and 6,413, disclosed in 952 (the receptors ligand activators of composition metal), composition with hprl receptor agonist activity, and G120RhGH, it has been human growth hormone (HGH) analog (Mode etc., 1966 of the effect of prolactin activator, Endocrinol.137 (2): 447-454), and as at United States Patent (USP) 5,506,107 and 5, the hprl receptor part of describing in 837,460. Also comprise prolactin GAP-associated protein GAP, S179D, people's prolactin and human placental lactogen.

PRL, GH and PL are the members of polypeptide hormone family, (summary is seen to have common structure, immunology and biological function, " Pancreatic Growth and Regeneration ", Ed.N.Sarvetnick, Ch.1.Brejie, T. etc., 1997), therefore be called PRL/GH/PL family here. PRL and GH are by vertebrate anterior pituitary secretion. In comprising osmotic adjustment, reproduction, lactation and immunoregulatory widely biological function, relate to PRL. Relevant physiology course occurs with growth and form and is correlated with in GH. Relevant receptors ligand is called as " PRL/GH/PL " receptors ligand. Based on the structural similarity of peptide and albumen, the functional similarity of the complementary aspect of gastrin, the functional similarity of one or more receptors bind aspect FACGINTs is divided into various group, each is organized all within the scope of various embodiments of the present invention. The hprl receptor part comprises PRL and PL, and the growth hormone receptor part comprises GH.

Any compound that comprises combination, interacts or stimulate the GLP-1 acceptor such as term used herein " GLP-1 receptors ligand ". The example of GLP-1 receptors ligand comprises GLP-1 and exendin-4. Glucagon-like-peptide-1 is synthetic with molecular forms GLP-1 (having the residue that routine is denoted as the 7-36 position) in enteroendocrine cell, and it is a kind of acid amides, and is similar with GLP-1 (7-37). The original research of GLP-1 BA is utilized the terminal extended pattern (1-37 and 1-36, the latter are a kind of acid amides) of the total length N-of GLP-1. Larger GLP-1 molecule lacks BA usually. Found afterwards to remove the first six amino acid and caused having the short GLP-1 molecule-type that BA increases considerably.

The major part of finding the GLP-1 that BA is arranged of circulation is GLP-1 (7-36) acid amide type, also can detect the bioactive GLP-1 of having of small amount (7-37) type. See Orskov, C. etc., Diabetes 1994,43:335-339. Two kinds of peptides all show the biological function of about same amount. GLP-1 replys nutrients digestion generation and secretes from enteroendocrine cell, plays multiple effect in the metabolism stable state after nutrient absorption. Cause the N end of peptide to degrade by what mediated by dipeptidyl peptidase (DPP-IV) the 2nd alanine residue cutting, the adjusting of GLP-1 occurs. DPP-IV is seen in general introduction. The BA of GLP-1 comprises stimulates dependence on the glucose insulin secretion and insulin biosynthesis, glucagon suppression secretion and gastric emptying, and suppresses food intake. As if GLP-1 have many additional effect in GI bundle and central nervous system, as at Drucker, and D., Endocrin 142:521-527, the review in 2001. Exemplary GLP-1 composition comprises: BIM 51077 (the GLP-1 analog of anti-DPP-IV digestion can obtain from Beaufour Ipsen); AC2592 (GLP-1, from Amylin, San Diego CA obtains); ThGLP-1 (GLP-1, the amino acid of modification and aliphatic acid attachment, from Theratechnologies, Saint-Laurent, Quebec, Canada obtains); CJC-1131 also is known as DAC^TM: GLP-1 (the GLP-1 analog of genetic engineering modified covalent coupling to the albumin, Conjuchem, Montreal, Quebec, Canada), (the GLP-1 analog of DDP-IV opposing is from Eli Lilly for the LY315902 of LY315902 and sustained release, Indianapolis, IN obtains); Low-molecular-weight GLP-1 analogies; Albugon (albumin: the GLP-1 fusogenic peptide, from Human Genome Sciences, Rockville, MD obtains); Liraglutide is also referred to as NN2211 (the long-acting GLP-1 derivative that the acidylate by the GLP-1 molecule obtains, in a single day it enter blood flow, with broad incorporation to preventing that it from being degraded by DPP-IV and reduce on the albumin that kidney removes, can be from Novo Nordisk, Denmark obtains; Elbrond etc., Diabetes Care 2002 Aug 25 (8): 1398-404).

Exendin-4, the example of exendin is the new peptide that obtains from Gila monster (Heloderma suspectum (Gila monster)) venom, with GLP-1 (7-36) acid amides 53% homology is arranged. Its function is the long-acting activator as glucagon-like peptide 1 (GLP-1) acceptor, because the degraded of its anti-DDP-IV. Exendin-4 has the character of similar GLP-1, regulates gastric emptying, insulin secretion, food intake and glucagon secretion. The example of exendin-4 comprises exenatide (synthesis type is also referred to as AC2993, Amylin); Exenatide LAR (long-acting type); ZP10 (adding the exendin-4 of the modification of 6 lysine residues, Aventis/Zealand Pharma); And AP10 (durative action preparation, Alkermes, Cambridge MA). Physiologic Studies is presented at the continuous expression of exendin-4 in the transgene mammal and does not upset glucose stable state, cell mass or food intake (Biaggio, L. wait .J Biol Chem 275:34472-34477,2000), therefore do not understand the physiological effect of exendin-4 fully.

DPP IV (DPP-IV) inhibitor refers to suppress the compound of DPP-IV activity, comprises 766 relevant peptases of amino acid whose film of GLP-1, GLP-2 and GIP in its substrate. The deactivation of the GLP-1 of DPP-IV mediation is the bioactive decisive factor of GLP-1 in the body. The example of DPP-IV inhibitor comprises isoleucine tetrahydro-thiazoles, valine-purrolidide, NVP-DPP738 (Novartis, Cambridge, MA), LAF237 (Novartis), P32/98 (Probiodrug AG, Halle, Germany) and P93/01 (Probiodrug).

Comprise the compound that stimulates the EGF acceptor such as term used herein " EGF receptors ligand ", so that when the gastrin/cck receptor in identical or adjacent tissue or the same individual is also stimulated, induce the pancreatic cell regeneration that produces insulin. The example of this EGF receptors ligand comprises length EGF, and it is EGF1-53, further comprises EGF1-48, EGF1-49, EGF1-52 and fragment and active analogue thereof. Other example of EGF receptors ligand is the TGF α type that comprises 1-48,1-47,1-51, and amphiregulin (amphiregulin) and poxvirus growth factor and proof and gastrin/cck receptor part have any EGF receptors ligand of identical synergistic activity. These comprise active analogue thereof, fragment and the modification of above-mentioned substance. Also see, Carpenter and Wahl, Chapter 4, in Peptide Growth Factors (Eds.Sporn and Roberts), Springer Verlag, 1990.

The compound group comprises further the EGF receptors ligand that comprises " modifying EGF ", and it comprises the variant of normal or wild type EGF. Verified should the modification affects one or more BAs, such as the clearance rate of EGF. This term comprises the peptide that has with the amino acid sequence of the amino acid sequence basic simlarity of people EGF, for example, has one or several 49-Phe ,82-Ser,115-Arg,144-Met,145-Asn ,161-Arg,169-Met Human Connective tissue growth factor in various residues position.

Restructuring EGF type is changed at structure and activity by genetic modification, for example, has described and has the EGF (U.S. Patent number 4,760,023) that replaces methionine on 21 with leucine. Recombined human EGF (hEGF) with 51 residues, that is, lack two C-terminal residues 52 and 53 of hEGF, and have neutral amino acid at 51 and replace, it is active to keep EGF, is more resisting proteasome degradation during the production process of microorganism and after giving the experimenter. The nucleic acid molecules (WO 00/29438) of the significantly similar protein family of a series of codings and EGF and TGF α has been described. The EGF mutain (EGF of sudden change) (WO 93/03757) that replaces the 16th residue histidine with neutrality or acidic amino acid has been described, the activity when this form is retained in low pH value. The chemical analog of EGF and TGF α and fragment keep the ability (U.S. Patent number 4,686,283) in conjunction with the various members of EGF receptor family. Useful performance is given in the various modifications of EGF or TGF α, affects one or more recombinant protein output, external and body internal stability and activity in vivo. The exemplary recombinant modified EGF receptors ligand that uses among the embodiment here is the C terminal deletion type of the people EGF of 51 amino acid longs, has asparagine (being called EGF51N here) at 51, and it keeps whole I.N.T. basically^TMActivity has in the body and/or vitro stability, and stability is at least approximately same high or than its higher (S.Magil etc., on May 15th, 2003 is with PCT/US02/33907, here in conjunction with its full content as a reference) with normal or wild type hEGF.

Term used herein " growth hormone " comprises with the total basic identical amino acid sequence of endogenous mammal growth hormone and has any polypeptide of mammal growth hormone biological activity. Human growth hormone (HGH) is polypeptide (Goeddel etc., 1979, the Nature 281:544-548 that contains 191 amino acid and the about 22k Dal of molecular weight in strand; Gray etc., 1985, Gene 39:247-254). This term comprises analog and the growth hormone of other species and the variant of natural generation that has disappearance, inserts or replace. See Cunningham etc., 1989, Science 243:1330-1336, and 1989, Science 244:1081-1085; And WO 90/05185, and U.S. Patent number 5,506,107.

Any endogenous mammalian EPO or its variant (EPO) such as term used herein " erythropoietin(EPO) ", perhaps EPO receptor stimulating agent, such as EPO analogies EMP1 (Johnson etc., 2000, Nephr Dial Tranpl 15:1274-1277); The analogies (Wrighton etc., 1996, the Science 273:458-464 that perhaps describe; U.S. Patent number 5,773,569; Kaushansky, 2001, Ann NY Acad Sci938:131-138); Have the EPO receptor agonist activity antibody (see, for example, U.S. Patent number 5,885,574; WO 96/40231); And at U.S. Patent number 6,333, the amino acid sequence of describing in 031 and 6,413,952.

Term described herein " PACAP " meaning is PACAP or its analog or the variant that endogenous produces, wherein analog or variant have basic identical or similar amino acid, BA (the Moro etc. that perhaps have PACAP receptor stimulating agent such as maxadilan, 1997, J Biol Chem 272:966-970. Useful PACAP variant includes but not limited to, such as the U.S. in the patent No. 5,128,242; 5,198,542; 5,208,320; With 6,242, disclosed 38 amino acid and 27 amino acid whose variants in 563).

Pharmaceutical composition

The present invention provides in various embodiments and comprises the FACGINT that treats effective dose and the pharmaceutical composition of gastrin/cck receptor ligand combination. Can prepare all pharmaceutical compositions described herein, contain or do not contain immunodepressant, contain or do not contain component or the device carried for sustained release, part or whole body. Can add pharmaceutically acceptable carrier or excipient. Such carrier includes but not limited to salt, buffer salt, glucose, water, glycerine, ethanol and combination thereof. Preparation should be fit to administering mode. Term used herein " effective dose " is to be enough to reach the therapeutic agent of generally acknowledged medical science terminal point or the amount of agent combination, in this example, generally acknowledges that the medical science terminal point is the correction of diabetic symptom. Those of skill in the art can determine effective dose by rule of thumb according to the accepted method of side phasing related parameter described herein.

The composition here can further comprise moistening or emulsifying agent, perhaps the pH buffer. Composition can be liquid solution, suspension, emulsion, tablet, pill, capsule, extended release preparation or pulvis. Composition can be formulated into suppository, contains traditional binding agent and carrier such as triglycerides. Oral formulations can comprise the carrier of standard, such as sweet mellow wine, lactose, starch, dolomol, saccharin sodium, cellulose, the magnesium carbonate of pharmacy level, etc. Various induction systems are known, and can be used for giving composition of the present invention, for example, seal liposome, particulate, microcapsules etc.

In exemplary embodiment, the composition will be here according to conventional steps is mixed with and is suitable for, and for example, is suitable for the subcutaneous people's of giving pharmaceutical composition. Typically, the composition of subcutaneous administration is to be dissolved in aseptic grade to open solution in the aqueous buffer solution. If necessary, composition also can comprise solubilizer and the local anaesthetics that improves injection site pain. Usually, component is independently or mixes with unit dosage form and provide, for example, and the drying in airtight container such as ampoule or sachette, cryodesiccated powder or without aqueous concentrate, for example, the amount of indication activating agent. When giving composition by transfusion, its available injection bottle that contains water, buffer solution or the salt solution of aseptic pharmacy level makes up a prescription. When giving composition by injection, the sterilized water that can be provided for injecting or the ampoule of salt solution are in order to can mix component before the administration. Here the composition that contains its various components can be mixed with suppository, it contains 0.5% active component to about 10% weight range of having an appointment; Oral formulations preferably contains 10% active component to about 95% weight of having an appointment.

Every daily dose perhaps is divided into many less fractionated doses with the single dose administration, several times administration in a day.

Composition of the present invention can be mixed with neutrality or salt form. The acceptable salt of pharmacy comprises those salt that form with free amine group, be derived from the salt of hydrochloric acid, phosphoric acid, acetic acid, tartaric acid etc. such as those, and the salt that forms of those and free carboxy, be derived from the salt of NaOH, potassium hydroxide, ammonium hydroxide, calcium hydroxide, iron hydroxide, isopropylamine, triethylamine, 2-ethamine ethanol, histidine, procaine etc. such as those.

To in the treatment of particular disorder or the state of an illness, will decide on the character of disorder or the state of an illness by effective therapeutic dose of the present invention, can measure by standard clinical techniques. The exact dose that uses in preparation also will be decided on the seriousness of method of administration and disease or disorder, and should determine according to doctor's judgement and every experimenter's situation. Blood insulin or C peptide level, and fasting glucose level or glucose brings out the conventional determining of level, is to measure by those of ordinary skills. Can pass through the area of pharmacology those of ordinary skill, infer effective dose from the dose-response curve that derives from external or animal model experiment system. The suitable dosage range of administration is every kind of active I.N.T. normally^TMAbout 0.01 microgram of compound per kilogram of body weight every day is to about 10,000 micrograms, and for example, about 0.01 microgram of per kilogram of body weight every day arrives about 10 milligram to about 10 micrograms, about 1 microgram to about 500 micrograms or about 10 micrograms to about 1 microgram, about 0.1 microgram. Therefore the suitable dosage range of administration is normally about 0.01 microgram/kg body weight/day to 10 milligram/kg body weight/day.

The present invention provides medicine bag or the kit that comprises one or more containers in another embodiment, and wherein vessel filling has the component of one or more pharmaceutical compositions of the present invention. In such bag or kit, can find to contain the container of in the gastrin of UD/cck receptor part and/or one or more FACGINT or the EGF receptors ligand every kind or both and one or more immunodepressant. In these components of medicine bag or kit one or more can be formulated into for sustained release or be used for being inserted into as an alternative thing of sustained release device, perhaps can be formulated into for the part and carry. With the associating of these containers can be various written materials, such as operation instruction, perhaps with the notice of government organs' defined form of production, use or the sale of management medicine or biological product, this notice reflection is ratified through mechanism that management is used for giving people's medicine production, use or sale. Kit or medicine Bao Keyu are embedded in the software associating in the computer-readable format in some embodiments.

Characteristics of the present invention are pancreas islet regenerative therapy (I.N.T. in one aspect^TM) composition and method, for example, gastrin and FACGINT, the binding immunoassay inhibitor stimulates the newly growth of β cell in the body, increases pancreas islet group, causes glucose-tolerant improvement in the diabetic subjects, for example, in the humans and animals of suffering from diabetes.

Gastrin/cck receptor part and EGF receptors ligand or FACGINT can give by single associating dosage, perhaps with the respectively administration of any order. " effectively uniting dosage " of these compositions is the dosage that causes that the fasting blood-glucose reduces or the insulin secretory cell amount increases or the blood insulin level increases or the β cell mass increases. Gastrin/cck receptor part in one embodiment, is that the residue on the long people's gastrin of 17 amino acid residues, 15 is leucine (1-17Leu15 is called G17 Leu15 here); In addition, the EGF receptors ligand be people EGF51N (S.Magil etc., on May 12nd, 2003 disclosed PCT/US02/33907, here in conjunction with its full content as a reference). Effective dose can comprise gastrin/cck receptor part to the ratio of EGF receptors ligand greater than 1, for example, effective dose comprise gastrin/cck receptor part to the ratio of EGF receptors ligand greater than 10. The approach that makes things convenient for that gives this dosage is systemic injection, for example, uses subcutaneous single-dose.

In another embodiment, use the reagent of Immunosuppression system to recipient subjects. For example, this reagent is low molecular weight organic compound, for example, is fujimycin 506, former times Luo Mosi, at least a shown in cyclosporin A and cortisone and the table 1 in the other medicines. In selective embodiment, this reagent is antibody, and for example, antibody is anti--CD11a antibody and also at other antibody shown in the table 1. In another selective embodiment, immunodepressant can be the antibody that the experimenter produces behind the immunization protocol, for example, and anti-GFAP or anti-S100 β. The experimenter may suffer from diabetes, and for example, the experimenter is the mouse that non-obese diabetic mouse (NOD mouse) or Streptozotocin are processed. The experimenter can be the people, for example, suffers from the diabetic subjects of I type or type ii diabetes, perhaps suffers from the prediabetes state or suffers from gestational diabetes mellitus or suffer from the past diabetes, for example, suffers from gestational diabetes mellitus in the gestation in the past.

In addition, estimating the β insulin secretory cell of recently generation or size and the function of pancreas islet is measurement standard physiology or Diagnostic parameters, comprising: beta Cell of islet group, beta Cell of islet quantity, beta Cell of islet percentage, blood-glucose, serum glucose, blood GH, pancreatic cell group, pancreatic beta cell quantity, fasting plasma C peptide content, serum insulin and pancreas insulin content.

Because diabetes are autoimmune diseases in some case, I.N.T.^TMEmbodiment be to the reagent of also using one or more Immunosuppression systems, namely, the experimenter of immunosuppressant treatment or experimenter, whole body is treated effective dose, for example, the receptors ligand of each among each among EGF and the gastrin/CCK or FACGINT and the gastrin/CCK is such as the combination of gastrin and EGF or the combination of FACGINT and gastrin/CCK.

Can use the treatment of many different endpoint determination gastrins and EGF or FACGINT, perhaps whether improve diabetes with the combination of gastrin and EGF or FACGINT and the treatment of immunodepressant, for example, improve functional β cell mass in the Islet allografts. These are included in behind β cell stimulatory agents such as glucose or the arginine injection mouse, measure the C-P of circulation and actrapid monotard's the increasing of blood plasma level; The reaction to gastrin/EGF treatment by the actrapid monotard's immunoreactivity that increases or the mRNA level of from Islet allografts, extracting proof; And the increase of the β cell quantity of measuring by the morphometry of pancreas islet in the treatment animal.

Term used herein " transplanting " meaning is by any generally acknowledged method in the prior art, perhaps according to method noted here, cell, tissue or organ composition is incorporated in the mammalian body. This composition is " graft ", and mammal is acceptor. Graft and acceptor can be homology, allogeneic or xenogenesis. Term used herein " from the body " meaning is cell, tissue or the organ that graft comes autoreceptor.

Also can by suffer from that Streptozotocin (streptozotocin) is induced or heredity (use is called the mouse species of non-obese diabetes or NOD) diabetes acceptor mouse in, the reverse reference islet beta cell function of hyperglycaemia strengthens. With gastrin, with EGF or FACGINT with after one or more immunosuppressant treatment diabetes recipient subjects, by cancelling the afterwards improvement of survival rate of insulin, and by correcting by the shown hyperglycaemia of fasting blood glucose levels, prove that the β cell function strengthens. In addition, observe pancreas insulin and the plasma C peptide both increases.

Immunosupress reagent and commercial source that table 1. is exemplary

Title	Company	Performance
			2-amino-1,3-propanediol derivative	Novartis	In the experimenter who accepts organ or tissue's allogeneic or allogeneic, be used for prevention or treatment chronic rejection
2-amino-2[2-(4-octyl phenyl) ethyl] propane-1,3-glycol hydrochloric acid	Yoshitomi   Pharmaceutical   Industries，Ltd	Immunosupress, the lymphocyte that suppresses to accelerate returns
			40-O-(2-ethoxy)-rapamycin, SDZ-RAD, everolimus (Everolimus) (Certican )	Novartis   Pharmaceuticals	Former times Luo Mosi (rapamycin) derivative is used for the acute kidney rejection; By suppressing rejection and the graft vasculopathy after cell proliferation reduces heart transplant
6-(3-dimethyl-amino propionyl) Forskolin (forskolin)	Matsumori Akia   Nippon Kayaju   Co Ltd	The immunosuppressive action that also is used for the treatment of autoimmune disease
			Ismipur (Purinethol , 6-MP)	Glaxo   SmithKline	Be used for the treatment of Crohn ' s disease, inflammatory bowel disease and be used for the organ transplant treatment
ABX-CBL(CBL-   1)	Abgenix	The anti-human T cell of target, B cell, NK cell and monocytic mouse monoclonal antibody are used for the treatment of the steroid graft versus host disease, the potential use in inflammatory and autoimmune disorder treatment
			Alefacept (people LFA-3IgG1 fusion, AMEVIVE )	Skin disease system/the BIOGEN of University of Utah	Destroy pathogenic memory T-lymphocyte; Be used for the treatment of psoriasis, the inflammatory disorder that a kind of T-is cell-mediated
HLA-B2702 peptide (Allotrap )	SangStat   Medical	People's peptide, effect and the cell-mediated toxicity of T-of blocking-up NK cell are used for for the first time kidney alloplast rejection of prevention

Antisense ICAM-1 inhibitor (ISIS 2302), Enlimomab, BIRR1, Alicaforsen)	ISIS-Boehringer   Ingleheim	Mouse monoclonal antibody, blocking leukocyte adhere to T-cell surface molecule (ICAM-1r); Treatment kidney transplant rejection
			Imuran (Imuran , Azasan )	Generic，Glaxo   SmithKline，   Prometheus   Laboratories，   aaiPharma	Treatment rheumatoid arthritis and prevention kidney transplant rejection, and other autoimmunity or inflammatory disorder are such as inflammatory bowel disease
BTI-322	MedImmune	The monoclonal antibody in the mouse source of target CD2 acceptor; Be used for preventing first kidney rejection, and the rejection for the treatment of resistance
			2-chlorodeoxyadenosine (Cladribine) (Leustatin )	Boehringer   Ingleheim	Antimetabolite and to the relatively selectable immunodepressant of lymphocyte; Be used for the treatment of the lymphatic system malignant tumour, for example, hairy cell leukemia
Endoxan (CTX, Neosar , Cytoxan , Procytox )	Generic	The immunodepressant for the treatment of of arthritis and the disorder of other autoimmunity and cancer
			Cyclosporin (cyclosporin A, cyclosporin) (Sandimmune , Neoral , SangCya )	Novartis	11 amino acid whose cyclic peptide; The blocking-up helper cell, the immunodepressant of in organ transplant treatment and other immunological diseases, using

Demethimmunomy cin " (L-683,742: also be described to 31-desmethoxy-31-hydroxyl-L-683,590)	Merck & Co	The treatment autoimmunity disease, infectious disease, and/or prevention of organ transplant rejection
			Dexamethasone (Decadron, Dexone, Dexasone))	Generic	AC, effective immunodepressant in various disorders
DHA (DHA)	There is not suitable company	Reduce the ratio of expressing CD4 or CD8 T cell, the immunodepressant of blocking-up antigen recognizing process; Taku etc., Journal of Agricultural and Food Chemistry, 2000; 48 (4): 1047
			FTY720 (oral myriocin (myriocin) derivative)	Novartis   Pharmaceuticals	Change lymphocytic infiltration to transplanted tissue; In kidney transplant, be used for the prevention organ rejection
Acetic acid Glatiramer (copolymer 1, Copaxone )	Teva   Pharmaceuticals	The synthetic peptide copolymer; The trick of simulation myelin structure is so that immunocyte is in conjunction with Copaxone rather than myelin; Be used for multiple sclerosis
			Glial fibrillary acidic protein (GFAP)	CalBiochem；   Synx Pharma	Having immunity in diabetes animal model transplants active; Winer etc., Nature Medicine 9:198 (2003)
Gusperimus (Gusperimus), (15-deoxyspergualin (15-deoxyspergualin)) (Spanidin )	Bristol Myers-   Squibb	The immunodepressant of intravenous injection; Suppress the generation of cytotoxic T cell, neutrophil and macrophage

Hul124 (anti--CD11 is a)	XOMA	Humanized monoclonal antibody; The CD11a acceptor of target T cell surface optionally suppresses the immune system rejection of transplant organ
			Infliximab (Remicade )	Centocor (subsidiary of Johson ﹠ Johnson)	Monoclonal antibody, in conjunction with and the deactivation humanTNF-α and; Be used for the treatment of Crohn ' s disease and rheumatoid arthritis
Interferon	Comprise Serono, the various companies of Biogen etc.	The immunological regulation performance
			ISAtx247	Isotechnika	Be used for the treatment of autoimmune disease such as rheumatoid arthritis and psoriasis
Isotretinoin (isotretinoin)		Immunodepressant reduces the T cell and the immunity challenge is produced the ability of replying and breeding. Vergelli etc., Immunopharmacology, 1997,31:191.
			Medi-500   (TIOB9)	MedImmune	The monoclonal antibody of target human T-cell's intravenous injection; The rejection for the treatment of acute kidney and graft versus host disease
Medi-507	The MedImmune/ biological implantation	Humanized antibody for the intravenous injection of anti-CD2T cell; Be used for the treatment of corticosteroid resistance graft to host disease and the rejection of prevention kidney
			Amethopterin (Rheumatrex , Amethopterin, Trexallo )	Wyeth   Lederle，Generic	The antimetabolite (and as cancer therapy drug) that is used for the treatment of sick, the serious psoriasis of Crohn ' s and adult's rheumatoid arthritis

Mitoxantrone (Mitoxantrone) (Novantrone )	Immunex	Anti-proliferative effect to the cell immune system that comprises T cell, B cell and macrophage; Be used for the treatment of difficult prostate cancer, acute myeloid leukaemia and multiple sclerosis with hormone therapy
			Mycophenolate (Mycophenolate mofetil) (CellCept )	Roche	The synthetic propagation that causes T and bone-marrow-derived lymphocyte by the blocking-up purine nucleotides; Be used for organ transplant and inflammatory bowel disease
OKT4A	The R.W.Johnson institute of materia medica	The mouse monoclonal antibody of the anti-human CD4 T of target cell; When with other immunosuppressive drug associating, be used for prevention kidney transplant rejection
			Muromonab-CD3   (Orthoclone   OKT3)	The R.W.Johnson institute of materia medica	Be attached to the monoclonal antibody on the φt cell receptor site, prevent transplanted tissue activation
Prednisolone (Prednisolone) (Deltasone , Oraone )		Corticosteroid suppresses the inflammation relevant with transplant rejection
			Basiliximab (basiliximab) (Simulect )	Novartis   Pharmaceuticals	Be attached to the monoclonal antibody of acceptor site on the T cell, prevent transplanted tissue (kidney transplant) activation
S100β	Neuroglian	In diabetes animal model, has immunosuppressive activity
			Former times Luo Mosi, rapamycin (Rapamune )	The Wyeth-Ayerst laboratory	Immunodepressant and the dependent cells factor (for example, IL-2) the potential inhibitor of T cell proliferation (kidney transplant)

Fujimycin 506 (Prograf (Prograf); FK-506)	Fujisawa	Disturb IL-2 TCR communication
			Antithymocyte immunoglobulin (ATGAM, Thymoglobulin )	SangStat Medical Corporation, Pharmacia and Upjohn	The Antithymocyte Globulin; Be used for the reverse of acute kidney transplant rejection and may be used for transplanting inductive treatment by off-label
Yi Lizuo monoclonal antibody (efalizumab) (Xanelim )	XOMA	By with endothelial cell surface on adhesion molecule interact, the T cell modulator of targeting T-cells, guiding T cell migration enters the activation of skin and guiding T cell; Be used for the treatment of psoriasis
			Daclizumab (Daclizumab) (Zenapax ), HAT (humanization is anti--Tac), SMART is anti--Tac, anti--CD25 and humanization be anti--the IL2-acceptor	Protein design laboratory/Roche	Monoclonal antibody by in conjunction with the IL-2 acceptor, suppresses IL-2 in conjunction with the IL-2 acceptor; The anti-alloplast of suppressor T cell (kidney transplant) activity

The reagent of any Immunosuppression reaction such as term used herein " immunodepressant " or " be used for immunosuppressant reagent " meaning. Exemplary immunodepressant is presented in the table 1, and any derivative of those reagent or function equivalent are considered to embodiment of the present invention of being fit to here and describing in the claims.

As used herein, dose form is showed any scheme of giving in the composition provided here, for example, consists of I.N.T.^TMThe combination of the component of composition or FACGINT and gastrin/cck receptor part, and one or more immunodepressant, every kind contains effective dose, simultaneously administration or administration in specified time interval each other, for example, within time interval of one day each other, perhaps as compound artifact, perhaps independently, comprise the amount of the composition that time per unit such as every day carry, and the duration of every kind of composition administration or time cycle.

Most insulin rely on diabetic subjects needs at least insulin injection every day. In the situation or the diet at diabetes management of certain disease, need several times of the every daily dose of insulin, result by frequent glucose monitoring indicates insulin administration, and another action need diabetic subjects is for example carried out 5 times the best management of disease every day.

Reduce by the fasting blood glucose levels, and reply level and the duration reduction that blood-glucose is increased by dietary sugars consumption challenge is produced, show because the pancreas islet regenerative therapy of the success of combined immunization inhibitor causes the alleviation of diabetes. In case the pancreas islet that succeeds regeneration, insulin administration reduces, and for example, reduces to 2 injections from 5 injections every day; Reduce to 1 injection from 2 injections every day; And from 1 time to not needing injection, according to the data indication that from the blood glucose levels of monitoring, obtains. When treatment during diabetic subjects, the those of ordinary skill in diabetes field, know after the fasting and under other physiological conditions, adjust insulin dose according to blood glucose levels.

For the known difference between species in normal data and the species, adjustment gives the dosage of experimenter's composition, normal data comprises the half-life dynamics, metabolism, secretion of absorption, distribution, circulation and the toxicologic standard of the receptors ligand in the embodiment here, for example, to every kind of primate and rodent species. Usually, dosage is adjusted to the amount that gives the rodent species and be higher than about 6 times to about 100 times that give the primate species amount.

Immunodepressant in the table 1 that gives to provide according to the manufacturer or other are equal to reagent, according to the weight standard of the known method of technical staff in the area of pharmacology to the experimenter. For example, usually by injection and the oral fujimycin 506 that gives, the common oral former times Luo Mosi that gives.

That the administering mode of receptors ligand composition and immunodepressant includes but not limited to is subcutaneous, in skin, sheath, in the intramuscular, peritonaeum, in the intravenous, nose and oral route. Can give compound by any conventional route, for example, by transfusion or single injection, by pump, by the absorption through epithelium or mucocutaneous lining (for example, oral mucosa, rectum and intestinal mucosa, etc.). Can unite one or more other bioactive agents and give the receptors ligand here. For example, if exist bacterium to infect, can give here composition and one or more antibiotic of recipient of method, if perhaps exist headache can give aspirin. Preferably give the receptors ligand here by the whole body approach.

The present invention provides the method by the combination treatment diabetes that give extended release preparation in one embodiment, wherein composition comprises gastrin/cck receptor part, for example, gastrin, and the EGF receptors ligand, perhaps FACGINT, for example, GLP-1, GH or prolactin are divided into ripe insulin secretory cell to affect the pancreas islet precursor. But whole body or local FACGINT and the gastrin/cck receptor part that gives in the composition.

Not limited by any special mechanism, give gastrin/cck receptor part, such as gastrin, and EGF receptors ligand or one or more members in the defined FACGINT group here, such as GLP-1, GH, or prolactin, obtain alone or in combination effective pancreatic cell regeneration time lengthening after the immunodepressant, according to method as described herein or by the known method of those of ordinary skill in known equivalent means or the area of pharmacology, with any or multiple sustained release that is mixed with in these receptors ligands or the factor or the reagent.

The present invention provides the method for prevention or treatment diabetes in total embodiment, the method comprises to its administration composition of needs, wherein said composition comprises EGF receptors ligand or FACGINT, such as extended release preparation at least a in GLP-1, exending-4, growth hormone, the prolactin, associating gastrin/cck receptor part, in EGF receptors ligand or FACGINT and gastrin/cck receptor part each is being enough to increase the amount of pancreas insulin secretion β cell quantity in the mammal, thereby treatment or prevention diabetes. Containing the treatment diabetic subjects of extended release preparation of FACGINT and experimenter's composition comprises: PTH GAP-associated protein GAP (PTHrP) receptors ligand such as PTHrP (PTHrP; Garcia-Ocana, A. etc., 2001, J.clin.Endocrin.Metab.86:984-988); HGF (HGF) receptors ligand such as HGF (HGF; Nielsen, J. etc., 1999, J Mol Med 77:62-66); Fibroblast growth factor (FGF) is such as FGF, and colloid forms Porcine HGF (KGF) receptors ligand such as KGF; Nerve growth factor (NGF) receptors ligand such as NGF; Gastrointestinal inhibitory peptide (GIP) receptors ligand such as GIP; Transforming growth factor β (TGF β) receptors ligand such as TGF β (disclosed U.S. Patent application 2002/0072115 on June 13rd, 2002), laminin receptor part such as EHS-laminin; Pancreas islet regeneration GAP-associated protein GAP (INGAP) receptors ligand such as INGAP; Bone morphogenetic factor (BMP) receptors ligand such as BMP-2; Vasoactive intestinal peptide (VIP) receptors ligand such as VIP; Glucogan-like peptide 1 receptor part such as GLP-1 and exendin-4, glucagon-like peptide 2 (GLP-2) receptors ligand such as GLP-2, and by being suppressed at the inhibitors of dipeptidyl IV (Hughes of enzyme remote-effects GLP-1 level related in its integrality, T.deng, 2002, Am Diabetes Assoc Abstract 272-or); REG receptors ligand such as REG albumen; Growth hormone (GH) receptors ligand such as GH, prolactin (PRL) receptors ligand such as PRL and human placental lactogen (PL); IGF (1 type and 2 types) receptors ligand such as IGF1 and IGF-2; Erythropoietin(EPO) (EPO) receptors ligand such as EPO (http: ∥ www. drinet.org/html/august_2002_.htm); β cytokine (member who also is considered to EGF family); Activin A receptors ligand such as activin A; The blood vessel activin A; VEGF (VEGF) receptors ligand such as VEGF; Bone morphogenetic factor (BMP) receptors ligand such as BMP-2; Vasoactive intestinal peptide (VIP) receptors ligand such as VIP; VEGF (VEGF) receptors ligand such as VEGF; Pituitary adenylate cyclase activated polypeptides (PACAP) receptors ligand such as PACAP; Granulocyte colony stimulating factor (G-CSF) receptors ligand such as G-CSF; Granulocyte macrophage colony stimulating factor (GM-CSF) receptors ligand such as GM-CSH; Platelet derived growth factor (PDGF) receptors ligand such as PDGF and secretin receptors ligand such as secretin. These extended release preparations also can comprise immunodepressant.

In one embodiment, whole body gives the extended release preparation of composition. Alternatively, the part, or the device or the method that are used for carrying the part give composition. Mammal is the diabetes mammals, and for example, mammal suffers from diabetes and has been at least about 1% of the mammal life-span. Usually, the amount of the extended release preparation of gastrin in the composition/cck receptor part, FACGINT or EGF receptors ligand is lower than the required least effective dose (LED) of blood-glucose in any independent reduction diabetes mammal in these materials basically. The amount that is enough to induce for a long time the pancreas islet precursor to be differentiated to form the insulin secretion islet cells that glucose replys to combine provides FACGINT or EGF receptors ligand and gastrin/cck receptor part.

Another embodiment of the invention provides the method for prevention or treatment diabetes, the method comprises being enough to improve the amount of pancreas islet precursor propagation in pancreatic tissue, the administration of using to needs comprises associating FACGINT or EGF receptors ligand, and the extended release preparation of the composition of gastrin/CCK receptors ligand, thereby treatment or prevention diabetes.

In yet another aspect, the invention provides the method for prevention or treatment diabetes, the method comprises that the administration of using to needs comprises associating FACGINT or EGF receptors ligand, and the extended release preparation of the composition of gastrin/cck receptor part, every kind to be enough to increase the amount of pancreas insulin secretion β cell quantity in the mammal; And the amount of measuring pancreas islet regeneration, thereby treatment or prevention diabetes. With give said composition before the blood-glucose measured compare, give said composition and reduced blood-glucose, for example, with give said composition before the blood-glucose measured compare, giving said composition, to reduce blood-glucose about 50%, perhaps about 70%. With give said composition before in the mammal measured GH concentration compare, GH concentration reduces. With give said composition before in the mammal measured serum insulin concentration compare, serum insulin concentration increases. With give said composition before in the mammal measured the pancreas insulin concentration compare, the pancreas insulin concentration increases.

On the other hand, the invention provides the method for inducing pancreas islet regeneration in the mammal, the method comprises the extended release preparation that comprises the composition of associating FACGINT or EGF receptors ligand and gastrin/cck receptor part to administration, every kind being enough to increase the amount of the propagation of pancreas islet precursor in the pancreatic tissue, thereby induce pancreas islet regeneration.

On the other hand, the invention provides the method for in mammal, inducing pancreas islet regeneration, the method comprises the composition of the extended release preparation that comprises associating FACGINT or EGF receptors ligand and gastrin/CCK receptors ligand, and every kind to be enough to increase the amount of pancreas insulin secretion β cell quantity in the mammal.

On the other hand, characteristic of the present invention is to comprise gastrin/cck receptor part, and the composition of FACGINT or EGF receptors ligand, with any extended release preparation that is mixed with in these reagent. Said composition contains effectively induces pancreas islet precursor propagation to form the dosage that ripe insulin secretory cell amount increases. In addition, composition contains the amount of effectively inducing the pancreas islet precursor to be differentiated to form the mature insulin secretory cell. Said composition can contain pharmaceutically acceptable carrier. Composition can comprise the reagent of Immunosuppression reaction.

In yet another aspect, the invention provides the kit for the treatment of or prevention diabetes, contain the composition that comprises gastrin/cck receptor part and FACGINT or EGF receptors ligand, wherein any all exists with the extended release preparation form, container, and operation instruction. Composition can comprise immunodepressant. The composition of kit can further comprise pharmaceutically acceptable carrier. The composition of kit can exist by unit dosage form.

Here the another embodiment of the invention that provides is the method that the stem cell in amplification and the diabetes acceptor of noble cells becomes insulin secretory cell, be included in transplanted cells in the acceptor, and containing each composition in the gastrin of effective dose/cck receptor part and FACGINT or the EGF receptors ligand, wherein one or more exist with the extended release preparation form. For example, transplanted cells is to obtain from the people. In addition, transplanted cells is to obtain from pancreas islet, umbilical cord, embryo or stem cell line. Usually, gastrin/cck receptor part is the hGastrin1-17Leu15. Usually, the EGF receptors ligand is EGF or TGF α, or the polypeptide substantially the same with EGF or TGF α respectively on the structure, has the biological function substantially the same with EGF or TGF α. In relevant embodiment, by being selected from following approach transplanted cells, for example, stem cell: be injected directly in the organ, and pass through intravenous administration. For example, with injection cell in the organ that is selected from pancreas, kidney and liver. Alternatively, use through skin or through the liver approach with cell delivery in portal vein. In another example, before transplanting, available set compound ex vivo treatment cell.

Here the another embodiment of the invention that provides is by measuring the method for the stem cell transplantation that reduces treatment people diabetes in the diabetes acceptor, the method comprises to acceptor uses in the extended release preparation of the gastrin of effective dose/cck receptor part and FACGINT or EGF receptors ligand each, with when shortage gives effective dose, transplant cells into the identical acceptor of other side and compare, the amount of cell reduces. Can further give the reagent of acceptor Immunosuppression system. For example, reagent is the chemicals that is selected from FK506, rapamycin (rapamycin), cyclosporin and cortisone. Alternatively, reagent is antibody, and for example, antibody is anti-CD4. In any method that relates to stem cell that provides, cell can obtain from the family member and preserve and be used for using later on before transplanting.

Embodiment of the present invention provide to be used with FACGINT that gastrin/the CCK part gives or Innovative method and the composition of EGF receptors ligand, and any or multiple sustained release that is mixed with in these reagent is used for the treatment of diabetes. The present invention provides following any extended release preparation in one embodiment: gastrin, associating FACGINT or EGF receptors ligand, with larger effect, usefulness and the effectiveness that obtains to be obtained with ACGINT or EGF receptors ligand than separately, perhaps any so that the direct bioavailability of whole preparations to be provided in these reagent of giving of associating. The invention provides with the preparation of direct bioavailable and compare, the treatment ratio of improved extended release preparation.

Use extended release preparation that the reduction of blood sugar is continued even the longer time.

The combination of gastrin in extended release preparation/cck receptor part and EGF receptors ligand or FACGINT, the whole body administration is than have larger effect when with the direct preparation administration of non-sustained release. Extended release preparation with gastrin/FACGINT combination or gastrin/EGF receptors ligand combination is observed the improvement of glucose-tolerant and the increase of pancreas insulin level.

The method for the treatment of diabetes comprises the extended release preparation of using composition to individuality in its individuality of needs, wherein said composition provides gastrin/cck receptor part and FACGINT, or gastrin/cck receptor part and EGF receptors ligand, composition contains is enough to affect the dosage that the pancreas islet precursor is differentiated to form the mature insulin secretory cell. The cell of differentiation is the pancreas islet precursor of hiding remaining in the pancreatic duct. Treatment insulin relies on diabetes, the method of I type or juvenile form diabetes particularly, comprise that preferred whole body gives the gastrin of diabetes mammal differentiation and regeneration amount/cck receptor part and FACGINT or gastrin/cck receptor part and EGF receptors ligand, wherein a kind of reagent or two kinds of reagent all are with the extended release preparation form, stimulate pancreas islet to regenerate to increase the quantity of functional glucose reactivity insulin excreting beta cell in the pancreas. The combination of gastrin and FACGINT or EGF receptors ligand, wherein any are with the extended release preparation form, will cause the pancreas islet regenerative response to be significantly higher than with identical reagent but observed to regenerative response with the direct preparation of non-sustained release. Exemplary gastrin/cck receptor part is gastrin or its synthetic gastrin derivative as described herein, and exemplary FACGINTs is GLP-1, GH and prolactin. Exemplary EGF receptors ligand is recombined human EGF, for example, EGF51N, it is a kind of that to have disappearance and 51 at C end be people's recombinant mutant EGF of 51 amino acid longs of asparagine.

Alternatively, use through skin or through the liver approach with cell delivery in portal vein. In another case, before the transplanting, available set compound ex vivo treatment cell.

The method of extended release preparation and local conveying and administration

With at least a being mixed with by sustained release or control release administration in gastrin/cck receptor part or EGF receptors ligand or the FACGINT reagent. Here and in the claims " sustained release " that use refers to material, device, preparation and/or gives at least a I.N.T.^TMThe combination of therapeutic agent produces continuously acceptor or discontinuously in time, as a certain amount of composition or at least a I.N.T. reagent periodically are provided, or immunodepressant. The time cycle of sustainable release is a few minutes, several hours, several days, a few week or or even some months. The release of active agent can be constant for a long time, and perhaps discharging can be long time period release. Release can be irrelevant with the state of an illness, perhaps produces to reply to the composition in the reaction environment or to other external event and can trigger release. For example, release can be by interior derived components such as insulin or glucose triggering, and perhaps discharging can be by the composition of external complement as reacting to administration and triggering.

Sustained release right and wrong extended release preparation is relative, wherein non-extended release preparation is moment or the dosage that very rapidly gives whole treatment reagent, but or after administration in very short time composition all be biological utilisation so that the major part of reagent arrives acceptor in the very short time. Basically moment bioavailable the dosage example comprise the aqueous solution of the reagent of parenteral, for example by single intraperitoneal injection, or oral, perhaps even by drip-feed administration in several hours. Therefore, the intravenous administration of cardiovascular preparation circulated in 15 seconds and spreads all over whole arterial system; But the drip-feed administration of anti-tumor agent comprising salmosin is complete biological utilisation in several seconds of the end of instiling. On the contrary, extended release preparation is by providing long-term bioavailability, and by avoid may be because the potential side effect that the initial high level of reagent cause be of value to recipient subjects. See Mathiowitz, E.,Encyclopedia of Controlled Drug Delivery.1999.New York：John Wiley。

Extended release preparation having been developed into whole body and part (perhaps target) that reagent is provided carries. Here various materials, device and the transport way and the L.N.T. that look back^TMReagent such as gastrin/cck receptor part, and EGF receptors ligand or FACGINT use together.

Obtained oral sustained release system, for example, contained the hard gelatin capsule of many type pillers, each type has the dressing of different thickness, the so not coated rapider release of some pillers. See Banga, A., Bus.Brief.:Pharmatech 2002:151-154. In the oral osmotic system, enter the osmotic pressure of the liquid generation core component of tablet by semipermeable membrane, it does not rely on the variation in the intestines and stomach. In order to protect polymer reactant, for example albumen such as growth factor, method comprise uses site specific conveying, protease inhibitors, carrier system or preparation as containing the hydrogel of polyacrylic acid backbone and bioadhesion performance.

Most protein reagents are by parenteral delivery, and the method for control protein parenteral delivery comprises that the polymer that uses based on lactic acid is such as poly-(D, L-lactide-co-glycolide; PLGA), it forms the biodegradable microsphere that core contains this reagent, and it is discharging this reagent above about 1 month time. In addition, can add liposome (below the seeing) Pegylation that polyethylene glycol (PEG) makes albumen or contains albumen by covalency.

Can be used for whole body through the skin conveying carries and local conveying. Following penetration enhancers can be carried to improve with unite use through the paster of skin, perhaps unites with the device that is used for ionotherapy (iontophoresis), ultrasonic wave importing (phonophoresis), mechanical osmosis (microporation) or with the electroporation that can wear electric installation.

Term used herein " the local conveying " refers to by approach and preparation or device or both administrations, so that basically treat specific target organ or tissue, and other Organ and tissue is not treated like this, and perhaps the therapeutic domain of target tissue or organ is accepted the dosage height accepted than non-target tissue or organ at least twice, at least 5 times or at least 10 times. Here common, target organ is pancreas. As shown here, can be to local cell or the many cells graft of carrying transplanting of pancreas in the portal vein by being expelled to; Can be by being expelled to delivering medicament in arteria pancreatica, arteria hepatica, portal vein or the pancreatic duct. Can use not implanted pump, that is, keep the outside, by the conduit that pump is connected on the organ delivery of composition be arrived target organ such as pancreas. Also may use the local delivering composition of implantable pump.

Local other method of carrying comprises and is not limited to endoscopic retrograde choledochopancreatography (ERCP); EUS guided fine needle suction biopsy (EUS-FNAD), the pharmaceutical composition that its suitable conveying provides here, rather than be used for sampling. For example see Wang etc., Transpl. Int 1995,8:268-272. Although these technology are designed to diagnose or be used for the purpose of prognosis, can they are revised the composition that is suitable for carrying here, it may be I.N. T. as described herein^TMComposition, with the immunodepressant associating, and or as extended release preparation. Also see Yano etc., 1994, Transpl Int, 7 Suppl 1:S187-193; Ricordi etc., 1994, Transpl Proc 26:3479; With Munoz-Acedo etc., 1995, J Endocrin 145:227-234.

Use the pump that is used for administration to treat several diseases, for example, be used for cancer and diabetes, wherein pump can be the pump of implantable or non-implantable (outside). Pump can be peristaltic pump, carbon fluorine boost pump or the osmotic pumps that comprises miniature osmotic pumps (Blanchard, S., 1996 Biomedical Engineering Applications, North Carolina State University). Peristaltic pump is along with the each time electric pulse of driving pump forefront is carried a series of amount medicines. Pump, electronic equipment and power supply are arranged in the titanium shell that covers silicon rubber. Medicament reservoir is to stand immense pressure, for example the silicone rubber capsule of 60psi. Can newly fill with medicament reservoir through tare weight by the polypropylene mouth. Carbon fluorine pump uses carbon fluorine liquid driving pump. Osmotic pumps uses osmotic pressure to discharge medicine with constant rate of speed. Exemplary pump is MiniMed MicroMed 407C pump (Medtronic, Inc., Northridge, CA). In addition, can use intrathecal drug delivery system (Medronic), it comprises two implantable elements, an infusion pump and a backbone inner catheter. Pump is inserted in the subcutaneous capsule at belly, and conduit is inserted into the sheath internal clearance of backbone, in the subcutaneous tunnel of doing, be connected on the pump. Then with constant or variable flow velocity administration. In addition, can use pump in the peritonaeum, implantable pump for example, the part, for example by conduit, perhaps whole body is carried the composition here.

Mucosa delivery to keep moist state and near below the upper dermatotome of vascular system may be that the organizational efficiency of dry epidermal surface is higher than passing through skin. Mucomembranous surface comprises: nose, lung, rectum, cheek, eye and genitals mucomembranous surface. Exemplary mucomembranous surface is nose and lung mucomembranous surface.

The material that is used for the microsphere of sustained release mainly is polymer, comprises PEG, is also referred to as PEO (PEO), and described PGLA. Polymeric excipient can directly be injected, and allows to be hydrolyzed gradually in subject or to degrade to discharge therapeutic agent. Can change polymer, for example, the molecular weight of PEG control release rate. The impact that PEG is not subjected to acceptor related in purge mechanism to Pegylation or the covalent attachment protected protein of protein for treatment agent is such as the acceptor of reticuloendothelial system (RES). Perhaps, can use polysaccharide with reagent target RES (United States Patent (USP) 5,554,386 that on September 10th, 1996 delivered). The organ of RES comprises liver, spleen and marrow.

Homopolymers or copolymer such as poly-(lactic acid-co-ethylene glycol) or PLA-PEG, can form the viscous liquid that mixes with the human cytokines agent. Molecular weight by PLA-PEG changes viscosity. Under certain conditions, in case be expelled in the subject therapeutic agent and polymer co-precipitation, and by diffusion forfeiture solvent, form like this storage (Whitaker, M. wait Bus.Brief:Pharnaatech 2002:1-5) with favourable release dynamics.

Form particulate by the polymeric microspheres that therapeutic agent is incapsulated. The polymer that uses in microsphere comprises poly-(lactic acid) or PLA; Poly-(glycolic) or PGA; And copolymer p LA-PGA. The amount of therapeutic agent such as albumen of the present invention, to discharge with the stage, as with the preliminary outburst of the albumen of the outside non-specific binding of particle, subsequent stage by diffusion, and the final stage by corroding, can be by the size of polymer composition, molecular weight, particulate, and physiological condition such as pH control. If albumen is with solid form, such as the freeze drying powder, the protein stability of microsphere production period improves, and it is with solvent or by cryospray method or ultrasonic processing method emulsification, then frozen suspension liquid further extracts solvent in liquid nitrogen. Microsphere can be from intermediary liquid, for example, and critical carbon dioxide (scCO₂) middle preparation.

Kumar, N. etc., Adv.Drug Deliv.Rev.53 (2001): 23-44 has described degradable bulky polymer and the synthetic method that is suitable for administration. Copolymer can be at random, replace perhaps block (double focusing or trimerization type), and configuration can be linear, perhaps star-like or grafting (graft) (comb shape). Polymer can form hydrogel, and it is the three-dimensional hydrophilic converging network that contains a large amount of waterborne liquids. By crosslinked or other chemical adduct so that the polymer that uses in the hydrogel do not dissolve.

Can from as at least a material that is selected from lower group prepare biodegradable implant: starch; Vinyl starch; DPG ester diacrylate (DPGDA); Tripropylene glycol ester diacrylate (TPGDA); Pectin; Acetylcellulose; Cellulose propionate; Cellulose acetate-butyrate; Cellulose-acetate propionate (CAP); Hydroxypropyl cellulose (HPC); Hydroxypropyl cellulose/cellulose-acetate propionate (HPC/CAP); Methyl methacrylate (MMA); Butyl methacrylate (BMA); Methylol methyl acrylate (HEMA); Acetic acid Octyl Nitrite (EHA); Octadecyl methacrylate (ODMA); And ethylene glycol dimethacrylate (EGDMA). See Gil, M. etc., Boletim de Biotecnologia 2002,72:13-19.

Except polymer, naturally occurring and synthetic lipid can be used for extended release preparation. DepoFoam^TMThe particle based on many bubbles lipid (liposome) during (Skye Pharma, London, Britain) formation incapsulates therapeutic agent (is seen United States Patent (USP) 5,993,850; And Ye, Q. etc., 2000J.Controlled Rel.64:155-166). Lipid has amphipathic, with net negative charge, and sterol, perhaps hybrid ion lipid and the method for preparing liposome are non-acid. The method that active treatment reagent is incorporated into liposome also is provided. Treatment reagent can be one or more among gastrin/cck receptor part, EGF receptors ligand and the FACGINT.

Other lipid of liposome is within the scope of the invention. Plant polarity liposome, for example ceramide such as wheat ceramide are used for forming gel with albumen such as alcohol soluble protein, one or more therapeutic agents wherein can be placed in one, and are used for through skin or mucosal. See the United States Patent (USP) 6,410,048 that on June 25th, 2002 delivered. Exemplary alcohol soluble protein comprises the wheat gliadin, and zeins. The polymer of other natural generation of using in the sustained release pharmaceutical preparation and device comprise collagen (EP-A-O 621 044), chitin (United States Patent (USP) 4,393,373), and the deacylated tRNA type of chitosan, chitin.

Penetration enhancers, for example, the ester of the fatty acid ester of glycolipid, non-esterified fatty acid, aliphatic alcohol, aliphatic alcohol, cyclohexanol, aliphatic acid, glycerine, ethylene glycol or aliphatic alcohol esters or ethylene glycol, be typical penetration enhancers, in transcutaneous device, can have other component such as stabilizing agent, solubilizer, surfactant and binder. See disclosed U.S. Patent application 20020127254 on September 12nd, 2002.

Use lipid and various types of polymer to be formed for " nanoparticle " of administration, summary is seen M.Kumar, 2002, J.Pharm.Pharmaceut.Sci.3:234-258. Discovery be filled into medicine in these particles maximum be the most lipophilic therapeutic agent. Have been found that with long-term lasting release of the liposome that comprises PLA, lecithin and phosphatid ylcholine or cholesterol.

Obtain local (target) sustained release by method described herein, for example, use the material by design target RES, or the liposome that is consisted of by the different materials of the cell of avoiding RES. Other targeted approach comprises uses antibody or in conjunction with the solvable recombinant receptor of liposome or microsphere outer surface. In addition, available any device as described herein such as pump, is further adjusted and is used extended release preparation described herein, is used for to specific target organ topical.

The present invention provides in another embodiment and comprises FACGINT or the EGF receptors ligand for the treatment of effective dose, and the sustained release pharmaceutical composition of gastrin/cck receptor ligand combination. Can add pharmaceutically acceptable carrier or excipient. Such carrier includes but not limited to salt solution, buffer salt, glucose, water, glycerine, ethanol and combination thereof. Preparation should be fit to administering mode.

Unless other definition is arranged, and all technology have the common identical implication of understanding with one skilled in the art of the present invention with scientific terminology here. In practice of the present invention, can use and those similar or be equal to method and materials as described herein. Described now the present invention comprehensively in various embodiments, illustrated extra embodiment by the following example and claim, this is not intended for use to be construed to further restriction. Therefore here in conjunction with the full content of the reference of all references as a reference.

Embodiment

Embodiment 1. usefulness GLP-1 receptors ligands, glucagon-like-peptide-1 (GLP-1), with Reach the treatment of gastrin/cck receptor part, gastrin, prevent from suffering from recently the NOD of diabetes The disease progression of mouse

Nonobese diabetes (NOD) is that mouse has the phenotype that has numerous disease pathogenesis feature with the human I type diabetes. The NOD mouse typically as far back as 4 the week age tamper indicating autoimmune pancreas inflammation of pancreatic islet and β cytoclasis. Onset diabetes occurs at the 10-of these mouse usually 15 ages in week, observe typical blood glucose levels and be at about 7mM to (comparing with the scope of about 3.0-6.6mM of normal mouse) between about 10mM, the pancreas insulin level reduces about more than 95% than normal mouse. Along with the progress of disease, the NOD mouse demonstrates more and more serious chronic diabetes sign, and blood glucose levels reaches about 25 between about 30mM, and the pancreas insulin level drops in fact and do not exist. In the serious stage of disease, destroyed above about 99% β cell.

In this embodiment, in recently suffering from the NOD mouse of diabetes, check the effect with GLP-1 and gastrin therapeutic alliance, measure and give serious hyperglycaemia, DKA and the dead and raising pancreas insulin content whether GLP-1 and gastrin will prevents recently to suffer from the NOD mouse of diabetes. The I.N.T. that uses^TMComposition is gastrin, is leucic synthetic people's gastrin I with 17 amino acid residues such as the 15th amino acids. The GLP-1 that uses is GLP-1, and it is that (processed precursor phase has residue than in the 7-36 position with this fragment for the bioactive fragment of people/mouse GLP-1; Obtain from Bachem H6795).

Monitor 12-14 age in week, the progress of Nonobese diabetes (NOD) female mice onset diabetes (fasting blood-glucose＞8.0 are to 15mmol/l), in 48 hours after symptom occurs, each group in two groups of mouse is carried out following treatment: only use vehicle treatment for one group; Another group gives 100 μ g/kg/ days GLP-1, and each treatment group is by approach twice administration every day in the peritonaeum.

Give treatment in 14 days. Monitor weekly fasting blood-glucose (FBG) level of animal. After removing food about 12 hours and behind last time peptide or vector injection, measured the FBG level in 24 hours. In case treatment stops, whether 4 weeks of FBG level (2-6 week) of monitoring again all mouse continue the prevention hyperglycaemia in order to determine after therapeutic treatment finishes. Treatment stopped at the 14th day, at the 18th day mouse was sampled to obtain FBG level (as shown in table 2).

The NOD mouse that diabetes are suffered from table 2. glucagon-like-peptide-1 (GLP-1) and the treatment of gastrin conjoint therapy recently

Group	GLP-1	Gastrin	Quantity	FBG (mM) (my god):
								0	7	14	18
				1   2   3   4	-   +   -   +	-   -   +   +	6   4   4   4					11.0±0.6   12.3±0.9   11.8±0.9   13.5±0.9	14.8±1.3   14.1±1.8   14.8±3.4   10.4±0.4	22.8±1.6   15.3±2.6   16.4±3.4   7.9±0.8	24.4±1.5   15.8±4.2   19.0±4.5   7.9±1.5

As implied above, by intraperitoneal injection, give every group NOD female mice GLP-1 (100 μ g/kg/s days) and gastrin (3 μ g/kg/s days) 2 every day. Use 12-14 age in week, in two days, begin to suffer from the diabetic mice of diabetes (it has been generally acknowledged that FBG surpasses 6.5mM).

This scheme is included in sampled for image data to these mouse in 6 weeks again, gathered blood and was used for measuring FBG and plasma C peptide, and put to death mouse assay pancreas insulin and pancreas islet inflammation (inflammation of pancreatic islet) is marked. From treatment at the beginning, mouse is neither accepted the insulin substitution treatment and is not also accepted immunosupress. Evaluation following parameters: the existence of survival rate, pancreas insulin level, pancreas islet inflammation and fasting blood glucose levels.

The result is presented in the animal of control group (group 1) of vehicle treatment, and fasting blood-glucose (FBG) value increased during this dummy treatment gradually, from the 0th day 18 days 24mM of 11mM glucose to the.

On the contrary, in the mouse with GLP-1 and gastrin Therapy, compare with the mouse (24.4 mM) of vehicle treatment, FBG value (7.9mM glucose) is reduction significantly, in fact is reduced to less than 1/3rd of in vehicle treatment mouse institute's generation level; Table 2). The most remarkable and the most surprisingly, being combined in of GLP-1 and gastrin reduced in the FBG level than separately with gastrin or use separately GLP-1 (the FBG level is respectively 19.0mM and 15.8mM) more effective. Only in the mouse with this combined therapy, FBG is reduced to the level of normal range (NR). The improvement that blood glucose levels is regulated in the mouse of GLP-1 and gastrin Therapy is considered to may be relevant with the remarkable increase of insulin content in the pancreas of these mouse.

In a word, the result shows with GLP-1 and gastrin low dosage the short of the mouse that recently suffers from diabetes is prevented from the progress of disease from reversing the normal approximately blood glucose levels of morbid state generation in this research. In addition, significantly being reduced in of this blood glucose levels continues one period after treatment stops again in the mouse of gastrin and GLP-1 treatment. Expectation is in these animals, and data will show that the pancreas insulin content increases, and shows that these impacts will stop rear last very long in treatment.

The NOD female mice of group is with hprl receptor part, prolactin (PRL) and gastrin/cck receptor part in addition, and with growth hormone receptor part, growth hormone (GH), and gastrin/cck receptor part, gastrin Therapy. Anticipatory data will show that these treatments are on FBG and other parameter generating impact similar to the data that obtain with GLP-1 treatment here.

Embodiment 2. contains or does not contain the comparison of Pegylation composition

Whether will provide than being formulated into for single-dose or non-sustained release administration in order to measure extended release preparation, the better effect of the identical preparation of other side compares with the I.N.T. that contains Pegylation or non-Pegylation^TMThe research of combination treatment NOD mouse scheme. I.N.T.^TMThe Pegylation of at least a component can prolong the time of the treatment reagent that keeps in vivo activity form in the composition.

Studies show that, and be mixed with directly, that is, and the I.N.T. of non-sustained release administration^TMComposition is compared, and gives I.N.T.^TMThe extended release preparation of at least a reagent can improve the diabetic disease states of NOD mouse in the composition.

The comparison of embodiment 3. dosage composition administration frequencies

Whether will provide than the better effect of the direct preparation of non-sustained release in order to measure extended release preparation, design comparison every day three times with give once a day NOD mouse I.N.T.^TMThe experiment of composition scheme.

The result shows, I.N.T.^TMThe bioavailability of composition prolongs, can with use I.N.T.^TMThe bioavailability that the extended release preparation of composition obtains is compared, and can improve effect, that is, and and with I.N.T.^TMDirect or the non-extended release preparation of composition is compared, and can correct better the symptom of the diabetic disease states of NOD mouse, and the frequency that can reduce such correction symptom required dosage.

Claims (110)

1. treat the method for diabetes, the method comprises the composition that comprises gastrin/cck receptor part and the factor (FACGINT) of replenishing the gastrin that is used for the pancreas islet regenerative therapy of the administration treatment effective dose of using to needs, as long as FACGINT is not the EGF receptors ligand.

2. according to claim 1 method, wherein FACGINT is selected from least a in lower group: the glucogan-like peptide 1 receptor part; The glucagon-like peptide 2 receptors ligand; Gastrointestinal inhibitory peptide (GIP) receptors ligand; Keratinocyte growth factor (KGF) receptors ligand; Inhibitors of dipeptidyl IV; REG protein receptor part; The growth hormone receptor part; Prolactin (PRL) receptors ligand; IGF (IGF) receptors ligand; PTH GAP-associated protein GAP (PTHrP) receptors ligand; HGF (HGF) receptors ligand; Bone morphogenetic protein (BMP) receptors ligand, transforming growth factor β (TGF-β) receptors ligand; The laminin receptor part; Vasoactive intestinal peptide (VIP) receptors ligand; Fibroblast growth factor (FGF) receptors ligand; The keratinocyte growth factor receptors ligand; Nerve growth factor (NGF) receptors ligand; Pancreas islet regeneration GAP-associated protein GAP (INGAP) receptors ligand; The activin A receptors ligand; VEGF (VEGF) receptors ligand; Erythropoietin(EPO) (EPO) receptors ligand; Pituitary adenylate cyclase activated polypeptides (PACAP) receptors ligand; Granulocyte colony stimulating factor (G-CSF) receptors ligand; Granulocyte macrophage colony stimulating factor (GM-CSF); Platelet derived growth factor (PDGF) receptors ligand; And secretin-receptor ligands.

3. according to claim 1 method, wherein FACGINT comprises the GLP-1 receptor part, it is GLP-1 or exendin-4.

4. according to claim 2 method, wherein FACGINT comprises the growth hormone receptor part, this growth hormone receptor part comprises growth hormone.

5. the method for the treatment of diabetes, the method comprises: a large amount of cells and the composition that comprises at least a FACGINT and gastrin/cck receptor part are exsomatized to be contacted, and needing only FACGINT is not the EGF receptors ligand; And give this cell of administration that needs it, thereby treatment diabetes.

6. according to claim 5 method, wherein cell is from body.

7. according to claim 1 or 5 method, wherein use or contact the composition that provides the amount of insulin secretory cell amount in effective increase mammal.

8. according to claim 1 or 5 method, wherein said composition is the whole body administration.

9. according to claim 1 or 5 method, wherein the amount of FACGINT reduces the minimum effective dose of the required FACGINT of blood-glucose in the diabetes mammal basically less than when lacking gastrin/cck receptor part in the composition.

10. according to claim 1 or 5 method, further comprise measuring and be selected from lower group parameter: the β cell mass that blood-glucose, serum glucose, blood GH, pancreatic beta cell group, serum insulin, pancreas insulin content and form tolerance are measured.

11. method according to claim 5, wherein cell is the pancreatic duct cell.

12. method according to claim 1 further comprises measuring being selected from lower group parameter: the amount of insulin secretory cell, the glucose response of insulin secretory cell, the amount of pancreas islet precursor propagation and the amount of mature insulin secretory cell.

Lead the method for regeneration 13. in mammal, induce pancreas, the method comprises being enough to increase the amount of pancreas islet precursor propagation in the pancreatic tissue, the composition that comprises the combination of FACGINT and gastrin/cck receptor part to administration, as long as FACGINT is not the EGF receptors ligand, thereby induce pancreas islet regeneration.

14. in mammal, induce the method for pancreas islet regeneration, the method comprises being enough to increase the amount of pancreas insulin secretion β cell quantity in the mammal, the composition that comprises the combination of FACGINT and gastrin/cck receptor part, wherein FACGINT is not the EGF receptors ligand.

15. method according to claim 5, further comprise contact procedure before, this cell of cultured in vitro.

16. comprise the composition of gastrin/cck receptor part and FACGINT, as long as FACGINT is not the EGF receptors ligand.

17. composition according to claim 16 contains the dosage of effectively inducing the pancreas islet precursor to be differentiated to form the mature insulin secretory cell.

18. composition according to claim 16 contains pharmaceutically acceptable carrier.

19. the kit for the treatment of or prevention diabetes contains the composition, container and the operation instruction that comprise gastrin/cck receptor part and FACGINT, as long as FACGINT is not the EGF receptors ligand.

20. kit according to claim 19, wherein said composition further comprises pharmaceutically acceptable carrier.

21. amplification and differentiated stem cells form the method for insulin secretory cell in the diabetes acceptor of transplanted cells, be included in and transplant stem cell in the acceptor, and use each composition among the gastrin that contains effective dose/cck receptor part and at least a FACGINT to acceptor, as long as FACGINT is not the EGF receptors ligand.

22. method according to claim 21, wherein this cell is to obtain from people or pig.

23. method according to claim 21, wherein transplanted cells is to obtain from pancreas islet, umbilical cord, embryo or stem cell line.

24. according to claim 1, the method for any one in 5,13,14 and 21, wherein gastrin/cck receptor part is gastrin.

25. method according to claim 24, wherein gastrin is Gastrin-17.

26. method according to claim 24, wherein gastrin/cck receptor part is people's Gastrin-17 Leul5.

27. method according to claim 23, wherein implanting cell in acceptor is to use the approach that is injected directly into organ and intravenous administration that is selected from.

28. method according to claim 23, wherein giving this cell is that the part is transported to the organ that is selected from pancreas, kidney and liver.

29. method according to claim 28, wherein local conveying cell is the step that is selected from lower group: endoscopic retrograde choledochopancreatography (ERCP); EUS guided fine needle administration (EUS-FNAD); Be expelled in the arteria pancreatica; Be expelled in the portal vein; Injection in the pancreas; And be expelled in the arteria hepatica.

30. method according to claim 27, wherein to inject cell be through skin or wear liver is transported to portal vein.

31. method according to claim 23, further be included in transplant step before, with composition ex vivo treatment cell.

32. reduce the method for the required transplanting for the treatment of people diabetes stem cell amount, the method comprises to acceptor uses among the acceptor gastrin of effective dose/cck receptor part and the FACGINT each, as long as FACGINT is not the EGF receptors ligand, wherein required cell concentration is compared when giving other same receptor effective dose in shortage, and the amount of required cell reduces.

33. method according to claim 1 and 2 further comprises the reagent of using the Immunosuppression reaction to the experimenter.

34. method according to claim 33, wherein the reagent of Immunosuppression reaction is medicine.

35. method according to claim 32, wherein the reagent of Immunosuppression reaction is selected from least a in lower group: rapamycin; Corticosteroid; Imuran; Mycophenolate; Cyclosporin; Endoxan; Amethopterin; Ismipur; FK506; DSG; FTY 720; Mitoxantrone; The 2-amino-1,3-propanediol; 2-amino-2[2-(4-octyl phenyl) ethyl] propane-1,3-glycol hydrochloric acid; 6-(3-dimethyl-amino propionyl) Forskolin; And demethimmunomycin.

36. method according to claim 32, wherein the reagent of Immunosuppression reaction is protein.

37. method according to claim 36, wherein protein comprises the amino acid sequence of antibody.

38. method according to claim 37, wherein the reagent of Immunosuppression reaction is selected from least a in lower group: hul 124; BTI-322; Allotrap-HLA-B270; OKT4A; Enlimomab; ABX-CBL; OKT3; ATGAM; Basiliximab; Daclizumab; Thymoglobuline; 1SAtx247; Medi-500; Medi-507; Alefacept; Yi Lizuo monoclonal antibody (efalizumab); Infliximab; And interferon.

39. method according to claim 32 wherein gives the reagent that pancreas islet regenerative therapy composition and Immunosuppression react in proper order.

40. method according to claim 32, wherein whole body gives at least a in pancreas islet regenerative therapy composition and the Immunosuppression reaction reagent.

41. method according to claim 40 wherein gives pancreas islet regenerative therapy composition as pill.

42. method according to claim 32 is wherein by being selected from intravenous, subcutaneous, the peritonaeum and the intramuscular approach gives at least a in the reagent of pancreas islet regenerative therapy composition and Immunosuppression reaction.

43. method according to claim 32, the wherein oral reagent that gives the Immunosuppression reaction.

44. method according to claim 32, wherein the reagent of Immunosuppression reaction is selected from least a in FK506, rapamycin and the daclizumab.

45. according to claim 1 or 32 method, wherein the experimenter is the people.

46. the kit for the treatment of diabetic subjects comprises immunodepressant, comprises the INT composition of FACGINT, as long as FACGINT is not the EGF receptors ligand, and container.

47. comprise the pharmaceutical composition of FACGINT and immunodepressant, as long as FACGINT is not the EGF receptors ligand.

48.I.N.T. ^TMThe pharmaceutical composition of the sustained release of therapeutic combination, said composition comprises: the gastrin-receptor part; With EGF receptors ligand or FACGINT; Wherein at least a among gastrin-receptor part or EGF receptors ligand or the FACGINT is extended release preparation.

49. according to claim 16 or 48 composition, further comprise immunodepressant.

50. composition according to claim 48, wherein extended release preparation is selected from Pegylation and based on the liposome of many bubbles lipid.

51. composition according to claim 48, wherein the EGF receptors ligand is selected from EGF and TGFA.

52. composition according to claim 48, wherein FACGINT is selected from least a in lower group: the glucogan-like peptide 1 receptor part; The glucagon-like peptide 2 receptors ligand; Gastrointestinal inhibitory peptide (GIP) receptors ligand; Keratinocyte growth factor (KGF) receptors ligand; Inhibitors of dipeptidyl IV; REG protein receptor part; The growth hormone receptor part; Prolactin (PRL) receptors ligand; IGF (IGF) receptors ligand; PTH GAP-associated protein GAP (PTHrP) receptors ligand; HGF (HGF) receptors ligand; Bone morphogenetic protein (BMP) receptors ligand, transforming growth factor β (TGF-β) receptors ligand; The laminin receptor part; Vasoactive intestinal peptide (VIP) receptors ligand; Fibroblast growth factor (FGF) receptors ligand; The keratinocyte growth factor receptors ligand; Nerve growth factor (NGF) receptors ligand; Pancreas islet regeneration GAP-associated protein GAP (INGAP) receptors ligand; The activin A receptors ligand; VEGF (VEGF) receptors ligand; Erythropoietin(EPO) (EPO) receptors ligand; Pituitary adenylate cyclase activated polypeptides (PACAP) receptors ligand; Granulocyte colony stimulating factor (G-CSF) receptors ligand; Granulocyte macrophage colony stimulating factor (GM-CSF); Platelet derived growth factor (PDGF) receptors ligand; And secretin-receptor ligands.

53. 1 composition according to claim 5, wherein the EGF receptors ligand is low-molecular-weight drug.

54. composition according to claim 48 is mixed with for parenteral.

55. composition according to claim 48 is mixed with for oral administration.

56. composition according to claim 50 is mixed with in, the peritonaeum subcutaneous be used to being selected from, the method for administration of intravenous and intramuscular injection.

57. composition according to claim 48 is wherein with at least a being mixed with for the whole body administration among gastrin-receptor part or EGF receptors ligand or the FACGINT.

58. according to claim 16 with 48 in any one composition, be mixed with for topical.

59. 8 composition according to claim 5, wherein topical is targeted pancreatic.

60. 8 composition wherein is selected from lower group method topical: endoscopic retrograde choledochopancreatography (ERCP) according to claim 5; The EUS guided fine needle attracts administration (EUS-FNAD); Be expelled in the arteria pancreatica; Be expelled in the portal vein; Injection in the pancreas; And be expelled in the arteria hepatica.

61. according to claim 16 with 48 in any one composition, be mixed be used to the method for administration that is selected from through skin and mucosa delivery.

62. pharmaceutical composition according to claim 48 is mixed with for passing through the mechanical device administration.

63. according to claim 16 with 48 in any one composition, be mixed with for being selected from following device administration: degradable implant; Transdermal patch; Conduit; Implantable pump; Through the skin pump; Infusion pump; And Iontophoretic device.

64. pharmaceutical composition according to claim 48 is mixed with in, the peritonaeum subcutaneous be used to being selected from, intravenous, and the method for administration in the pancreas.

65. 4 pharmaceutical composition according to claim 6, wherein intravenous route is to be transported to portal vein.

66. 2 pharmaceutical composition according to claim 6, wherein device is pump.

67. 2 pharmaceutical composition wherein is topical according to claim 6.

68. composition according to claim 48, wherein administration is topical and by being selected from following administration: endoscopic retrograde choledochopancreatography (ERCP); The EUS guided fine needle attracts administration (EUS-FNAD); Be expelled in the arteria pancreatica; Be expelled in the portal vein; Injection in the pancreas; And be expelled in the arteria hepatica.

69. 2 pharmaceutical composition wherein is the whole body administration according to claim 6.

70. pharmaceutical composition according to claim 48 contains effective dose.

71. comprise the kit of at least a dosage of composition according to claim 48.

Use I.N.T. 72. reduce^TMThe method of the frequency of combination treatment diabetic subjects, the method comprises: at least a component in the composition is made extended release preparation; And use said composition according to scheme to the experimenter, this scheme is not than being that like this preparation and the identical composition of other side have the longer time interval between treatment.

73. 2 method according to claim 7 is wherein by being selected from lower group administration: endoscopic retrograde choledochopancreatography (ERCP); The EUS guided fine needle attracts administration (EUS-FNAD); Be expelled in the arteria pancreatica; Be expelled in the portal vein; Injection in the pancreas; And be expelled in the arteria hepatica.

74. I.N.T. in the raising diabetic subjects^TMThe method of composition effect, the method comprises: use at least a component in the composition to the experimenter and be formulated into I.N.T. for generation of sustained release^TMComposition; And relatively give said composition amount treatment experimenter's effect and do not have the component of such preparation and the effect of the identical composition of other side, the amount that reduces or eliminates the required sustained release reagent preparation of experimenter's diabetic symptom by mensuration reduces, so that have the I.N.T. of sustained release compositions formulated^TMThe effect of composition improves.

75. 4 method according to claim 7, wherein relatively effect is the toxicity of further analysed composition so that with the component that does not have such preparation and the identical I.N.T. of other side^TMThe toxicity of composition is compared, give composition after less or slighter harmful symptom show to have at least a set of dispense and make the toxicity of the composition that produces sustained release and reduce.

76. the method for any one among the 3-75 according to claim 7, wherein the extended release preparation of component is selected from Pegylation and based on the liposome of many bubbles lipid.

77. the method for any one among the 3-76 according to claim 7, the component that wherein has extended release preparation is the EGF receptors ligand that is selected from EGF and TGF α.

78. the method for any one among the 3-76 according to claim 7, wherein FACGINT is selected from least a in lower group: the glucogan-like peptide 1 receptor part; The glucagon-like peptide 2 receptors ligand; Gastrointestinal inhibitory peptide (GIP) receptors ligand; Keratinocyte growth factor (KGF) receptors ligand; Inhibitors of dipeptidyl IV; REG protein receptor part; The growth hormone receptor part; Prolactin (PRL) receptors ligand; IGF (IGF) receptors ligand; PTH GAP-associated protein GAP (PTHrP) receptors ligand; HGF (HGF) receptors ligand; Bone morphogenetic protein (BMP) receptors ligand, transforming growth factor β (TGF-β) receptors ligand; The laminin receptor part; Vasoactive intestinal peptide (VIP) receptors ligand; Fibroblast growth factor (FGF) receptors ligand; The keratinocyte growth factor receptors ligand; Nerve growth factor (NGF) receptors ligand; Pancreas islet regeneration GAP-associated protein GAP (INGAP) receptors ligand; The activin A receptors ligand; VEGF (VEGF) receptors ligand; Erythropoietin(EPO) (EPO) receptors ligand; Pituitary adenylate cyclase activated polypeptides (PACAP) receptors ligand; Granulocyte colony stimulating factor (G-CSF) receptors ligand; Granulocyte macrophage colony stimulating factor (GM-CSF); Platelet derived growth factor (PDGF) receptors ligand; And secretin-receptor ligands.

79. 8 method according to claim 7, wherein component is low-molecular-weight drug.

80. the method for any one among the 3-76 according to claim 7, wherein administration is to carry by the approach that is selected from lower group: parenteral, oral, in skin, subcutaneous, mucous membrane, peritonaeum, in the intravenous, pancreas and intramuscular.

8l. 0 method according to claim 8, wherein administration produces local allocation.

82. 2 method further comprises the composition that gives effective dose according to claim 8.

83. 0 method wherein before administration, is mixed with the suitable sustained release device that uses with composition according to claim 8.

84. 3 method according to claim 8, wherein device is selected from lower group: degradable implant; Transdermal patch; Conduit; Implantable pump; Through the skin pump; Infusion pump; And Iontophoretic device.

85. 4 method according to claim 8, wherein device is pump.

86. 0 method is to be expelled in the portal vein by the intravenous route administration wherein according to claim 8.

87. reduce the method for the frequency for the treatment of diabetic subjects, the method comprises preparing uses I.N.T. by long-term continuous release to the experimenter^TMThe device of composition; Provide this device to the experimenter; And by displacement or refill device and repeatedly treat the experimenter.

88. 7 method according to claim 8, wherein device is pump.

89. 8 method according to claim 8, wherein pump is selected from lower group: through the skin pump; Carbon fluorine (flurorcarbon) propellant pump; Osmotic pumps; Miniosmotic pump; Implantable pump; And infusion pump.

90. 7 method according to claim 8, wherein device is to be selected from lower group: degradable implant; The implant of non-degraded; The mucosal adhesive implant; Transdermal patch; Conduit; And Iontophoretic device.

91. amplification and differentiated stem cells form the method for insulin secretory cell in the diabetes acceptor of cell, comprising: transplanted cells in acceptor; And comprise the gastrin of effective dose/cck receptor part; With each the sustained-release composition in FACGINT or the EGF receptors ligand, amplification and differentiated stem cells formation insulin secretory cell in acceptor wherein.

92. the composition for the treatment of diabetes comprises glucagon-like-peptide-1 (GLP-1) receptors ligand and gastrin/cck receptor part.

93. 2 composition according to claim 9, wherein the GLP-1 receptors ligand is GLP-1.

94. the composition for the treatment of diabetes comprises growth hormone (GH) receptors ligand and gastrin/cck receptor part.

95. 4 composition according to claim 9, wherein GH is human growth hormone (HGH) (HGH).

96. the composition for the treatment of diabetes comprises prolactin (PL) receptors ligand and gastrin/cck receptor part.

97. 6 composition according to claim 9, wherein PL is people PL.

98. the composition of any one among the 2-97 according to claim 9, wherein gastrin is that the 15th amino acids is 17 the amino acid whose gastrins that have of leucine residue.

99. the composition of any one among the 2-98 further comprises immunodepressant according to claim 9.

100. the composition of any one among the 2-99 further is mixed with for sustained release according to claim 9.

101. the method for the treatment of diabetic subjects comprises to the experimenter and uses the composition that comprises gastrin/cck receptor part and glucagon-like-peptide-1 (GLP-1) receptors ligand.

102. the method for the treatment of diabetic subjects comprises to the experimenter and uses the composition that comprises gastrin/cck receptor part and growth hormone (GH) receptors ligand.

103. the method for the treatment of diabetic subjects comprises to the experimenter and uses the composition that comprises gastrin/cck receptor part and prolactin (PL) receptors ligand.

104. the method for any one among the claim 101-103 further comprises giving immunodepressant.

105. the method for any one among the claim 101-104 further comprises and uses sustained release device to give at least a in receptors ligand or the reagent.

106. the method for any one among the claim 101-104 further comprises at least a being mixed with for sustained release in receptors ligand or the reagent.

107. the method for any one among the claim 101-104, wherein diabetic subjects has type i diabetes or type ii diabetes.

108. enlarge the method for the functional β cell mass of Islet allografts in the diabetic subjects acceptor of the Islet allografts of purifying, the method comprises gastrin cck receptor part and the FACGINT to the administration effective dose.

109. the method for the treatment of people diabetes comprises the pancreas islet goods is transplanted to diabetic subjects; And use the gastrin of effective dose/cck receptor part and FACGINT to the experimenter.

110. method according to claim 1, wherein FACGINT comprises that it is the hprl receptor part of prolactin.