EP1356295A2 - Test de diagnostic de polypeptide inhibiteur gastrique destine a la detection de la sensibilite aux diabetes de type 2, a une intolerance au glucose, ou a une glycemie a jeun - Google Patents

Test de diagnostic de polypeptide inhibiteur gastrique destine a la detection de la sensibilite aux diabetes de type 2, a une intolerance au glucose, ou a une glycemie a jeun

Info

Publication number
EP1356295A2
EP1356295A2 EP01930773A EP01930773A EP1356295A2 EP 1356295 A2 EP1356295 A2 EP 1356295A2 EP 01930773 A EP01930773 A EP 01930773A EP 01930773 A EP01930773 A EP 01930773A EP 1356295 A2 EP1356295 A2 EP 1356295A2
Authority
EP
European Patent Office
Prior art keywords
gip
individual
diabetes
type
glucose
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Withdrawn
Application number
EP01930773A
Other languages
German (de)
English (en)
Inventor
Michael A. Nauck
Juris J. Meier
Katrin Huecking
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
BioNebraska Inc
Original Assignee
BioNebraska Inc
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by BioNebraska Inc filed Critical BioNebraska Inc
Publication of EP1356295A2 publication Critical patent/EP1356295A2/fr
Withdrawn legal-status Critical Current

Links

Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
    • G01N33/6893—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids related to diseases not provided for elsewhere
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/435—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans
    • G01N2333/575—Hormones
    • G01N2333/62—Insulins
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2800/00—Detection or diagnosis of diseases
    • G01N2800/04—Endocrine or metabolic disorders
    • G01N2800/042—Disorders of carbohydrate metabolism, e.g. diabetes, glucose metabolism

Definitions

  • the present invention relates to methods and kits for determining whether an individual is susceptible to developing Type-2 diabetes, impaired glucose tolerance (IGT), or impaired fasting glucose (IFG). These methods and kits are used to measure the response of an individual to administration of gastric inhibitory polypeptide (GIP) or a GIP variant, thereby determining whether the individual is at risk for developing Type-2 diabetes, IGT, or IFG.
  • GIP gastric inhibitory polypeptide
  • Type-2 diabetes is believed to account for approximately 90-95% of all diabetes cases. There are currently no reliable diagnostic tests for determining if an individual is susceptible to developing Type-2 diabetes.
  • IGT is also common in the U.S. population, with an estimated 13.4 million Americans having this condition. IGT describes a condition where an individual has a reduced ability to tolerate glucose administration but the level of impairment is below that of definitive diabetes. Persons with IGT are at an increased risk of developing Type-2 diabetes as well as other diseases. IFG is a condition similar to IGT. IFG is also a prevalent in the U.S. and elsewhere. There are currently no reliable diagnostic tests for determining if an individual is susceptible to developing IGT or IFG.
  • GIP plays a role in the pathology of Type-2 diabetes based on data showing that the insulinotropic effects of GIP are significantly reduced in type-2 diabetes patients (7).
  • the precise role of GIP in the pathology of Type-2 diabetes remains obscure and prior to the present invention a link between GIP and an individual's susceptibility to developing Type-2 diabetes, IGT, or IFG had not been discovered.
  • the present invention is directed to, for example, methods and kits for determining whether an individual is susceptible to developing impaired glucose tolerance (IGT), impaired fasting glucose (IFG), or Type-2 diabetes.
  • the methods include administering a GIP or GIP variant to an individual, measuring the response of the individual and determining whether the individual is susceptible to developing IGT, Type-2 diabetes, or IFG.
  • Fig. 1 Plasma concentrations of glucose (capillary measurement; A) and immunoreactive GIP (B) in 21 first-degree relatives of type-2 diabetic patients (filled diamonds), 10 type-2 diabetic patients (open circles), and 10 healthy control subjects (filled circles) participating in hyperglycemic clamp experiments with intravenous infusions of GIP (2 pmol-kg "1 -min 1 ). Mean ⁇ SEM. P values: repeated-measures ANOVA (A: between subject/patient groups; B: with time; AB: interaction of group and time). *: significant difference (p ⁇ 0.05) to Type-2 diabetic patients; f: significant difference (p ⁇ 0.05) to normal subjects (Student's t-test)
  • FIG. 2 Plasma concentrations of insulin (A) and C-peptide (B) and insulin secretion rates (C) in 21 first-degree relatives of Type-2 diabetic patients (filled diamonds), 10 Type-2 diabetic patients (open circles), and 10 healthy control subjects (filled circles) participating in hyperglycemic clamp experiments with intravenous infusions of GIP (2 pmol-kg "1 -min "1 ). Mean + SEM. P values: repeated-measures ANOVA (A: between subject/patient groups; B: with time; AB: interaction of group and time). *: significant difference (p ⁇ 0.05) to Type-2 diabetic patients; f: significant difference (p ⁇ 0.05) to normal subjects (Student's t-test). In the right panels, individual plasma concentrations of insulin (D) and C-peptide (E) and insulin secretion rates (F) in 21 first-degree relatives are shown in relation to the upper and lower 95 % confidence interval for normal subjects (dashed lines).
  • Fig. 3 Differences ( ⁇ ) between the mean values at time points 15 and 30 min (“hyperglycemia") and 75 and 90 min ("hyperglycemia plus exogenous GIP) for insulin (A) and C-peptide (B) concentrations and for insulin secretion rates (C) in 21 first-degree relatives of Type-2 diabetic patients (filled diamonds), 10 Type-2 diabetic patients (open circles), and 10 healthy control subjects (filled circles). P values: ANOVA (overall comparison) and Student's t-test (comparison of individual groups).
  • Fig. 4 Plasma concentrations of glucagon in 21 first-degree relatives of
  • Type-2 diabetic patients filled diamonds
  • 10 Type-2 diabetic patients open circles
  • 10 healthy control subjects filled circles
  • intravenous infusions of GIP 2 pmol-kg "1 -min "1 ).
  • P values repeated-measures ANOVA (A: between subject/patient groups; B: with time; AB: interaction of group and time). *: significant difference (p ⁇ 0.05) to Type- 2 diabetic patients; t: significant difference (p ⁇ 0.05) to normal subjects (Student's t-test).
  • Fig. 5 Plasma concentrations of proinsulin in 21 first-degree relatives of Type-2 diabetic patients (filled diamonds), 10 Type-2 diabetic patients (open circles), and 10 healthy control subjects (filled circles) participating in hyperglycemic clamp experiments with intravenous infusions of GIP (2 pmol-kg "1 -min 1 ). Mean + SEM. P values: repeated-measures ANOVA (A: between subject/patient groups; B: with time; AB: interaction of group and time). *: significant difference (p ⁇ 0.05) to Type-2 diabetic patients; f: significant difference (p ⁇ 0.05) to normal subjects (Student's t-test).
  • Impaired glucose tolerance refers to an intermediate stage between normal (an individual without impaired glucose metabolism) and definitive or “frank” diabetes.
  • impaired glucose tolerance is defined as a two-hour value between 140 mg per dL (7.8 mmol per L) and 199 mg per dL (11.0 mmol per L).
  • Impaired fasting glucose refers to a condition similar to IGT.
  • paired fasting glucose is defined as a fasting plasma glucose concentration between 110 mg per dL (6.1 mmol per L) and 125 mg per dL (6.9 mmol per L).
  • Type-2 diabetes also referred to as non-insulin-dependent diabetes mellitus or adult-onset diabetes
  • non-insulin-dependent diabetes mellitus or adult-onset diabetes is a classification used to describe individuals who exhibit insulin resistance and who usually exhibit relative, rather than absolute, insulin deficiency. The precise cause of Type-2 diabetes has not been identified.
  • Illustrative, but non-limiting criteria for determining whether an individual has Type-2 diabetes include one or more of the following: (1) a confirmed fasting plasma glucose value of greater than or equal to 126 milligrams/deciliter (mg/dL), (2) in the presence of symptoms of diabetes, a confirmed non-fasting plasma glucose value of greater than or equal to 200 mg/dL (3) with an oral glucose tolerance test (by administering 75 grams of anhydrous glucose dissolved in water, in accordance with World Health Organisation standards, and then measuring the plasma glucose concentration 2 hours later), a confirmed glucose value of greater than or equal to 200 mg/dL.
  • mg/dL milligrams/deciliter
  • IGT, IFG, and Type-2 diabetes are those proposed in 1997 by the "Expert Committee on the Diagnosis and Classification of Diabetes Mellitus.” See Expert Committee on the Diagnosis and Classification of Diabetes Mellitus, Diabetes Care 20: 1183 (1997). It is understood, however, that the criteria for diagnosing IGT, IFG, and Type-2 diabetes are set by institutional bodies and may be changed from time to time and may vary from organisation to organisation. Notwithstanding these variations, the terms IGT, IFG, and Type-2 diabetes, as used herein, are intended to be interpreted broadly and to be inclusive of the varying classification criteria used in the art. "Susceptible to developing IGT, IFG, or Type-2 diabetes" means at risk for developing one or more of these conditions.
  • the present invention can be used to assess individuals who already exhibit one or more risk factors of developing these diseases.
  • the inventive methods are applied to an individual who is a first-degree relative of a Type-2 diabetes patient. In this case, the individual is already in a high risk category, from an epidemiological standpoint.
  • the present invention provides such a predictive diagnostic tool.
  • GIP glucose-dependent insulinotropic hormone
  • the GIP used is synthetic human GIP, having the following amino acid sequence of human GIP: Tyr-Ala-Glu-Gly-Thr-Phe-Ile-Ser-Asp-Tyr-Ser-Ile-Ala-Met-Asp-Lys-Ile- His-Gln-Gln-Asp-Phe-Val-Asn-Trp-Leu-Leu-Ala-Gln-Lys-Gly-Lys-Lys-Asn-Asp-Trp- Lys-His-Asn-Ile-Thr-Gln (SEQ ID NO: 1).
  • the present invention includes the use of recombinant human GIP as well as GIP derived from other species, whether recombinant or synthetic.
  • biologically active variants of GIP include biologically active variants of GIP.
  • Biologically active in this context, means having GIP biological activity, but it is understood that the activity of the variant can be either less potent or more potent than native GIP.
  • GIP biological activity can be determining by in vitro and in vivo animal models and human studies as is well known to the skilled artisan.
  • GIP variants are any molecules, whether they be peptides, peptide mimetics, or other molecules that bind to or activate the GIP receptor and its second messenger cascade. The GIP receptor has been characterised in the art.
  • GIP peptides containing one or more amino acid substitutions, additions or deletions.
  • the number of substitutions, deletions, or additions is 30 amino acids or less, 25 amino acids or less, 20 amino acids or less, 15 amino acids or less, 10 amino acids or less, 5 amino acids or less or any integer in between these amounts.
  • the substitutions include one or more conservative substitutions.
  • a "conservative" substitution denotes the replacement of an amino acid residue by another, biologically active similar residue.
  • conservative substitution examples include the substitution of one hydrophobic residue, such as isoleucine, valine, leucine or methionine for another, or the substitution of one polar residue for another, such as the substitution of arginine for lysine, glutamic for aspartic acids, or glutamine for asparagine, and the like.
  • substitution of one hydrophobic residue such as isoleucine, valine, leucine or methionine for another
  • substitution of one polar residue for another such as the substitution of arginine for lysine, glutamic for aspartic acids, or glutamine for asparagine, and the like.
  • GIP variant is a polypeptide comprising amino acids 19-30 of SEQ ID NO: 1 (see Morrow et al., Canada J. Physiol Pharmacol. 74:65 (1996)), where 1-5 or more amino acid substitutions, deletions, or additions can be made within this 19-30 amino acid sequence.
  • GIP variants include the above described peptides which have been chemically derivatized or altered, for example, peptides with non-natural amino acid residues (e.g., taurine residue, beta and gamma amino acid residues and D-amino acid residues), C-terminal functional group modifications such as amides, esters, and C-terminal ketone modifications and N-terminal functional group modifications such as acylated amines, Schiff bases, or cyclization, such as found for example in the amino acid pyroglutamic acid.
  • non-natural amino acid residues e.g., taurine residue, beta and gamma amino acid residues and D-amino acid residues
  • C-terminal functional group modifications such as amides, esters
  • C-terminal ketone modifications such as acylated amines, Schiff bases, or cyclization, such as found for example in the amino acid pyroglutamic acid.
  • sequence identity refers to a comparison made between two molecules using standard algorithms well known in the art.
  • the preferred algorithm for calculating sequence identity for the present invention is the Smith-Waterman algorithm, where SEQ ID NO: 1 is used as the reference sequence to define the percentage identity of polynucleotide homologs over its length.
  • the choice of parameter values for matches, mismatches, and inserts or deletions is arbitrary, although some parameter values have been found to yield more biologically realistic results than others.
  • Exhibits at least one indicia of susceptibility for developing Type-2 diabetes, IFG, or IGT refers to epidemiological risk factors for developing one or both of these diseases.
  • risk factors include, but are not limited to, older age, particularly, 45 years or older, obesity, family history of diabetes, particularly, individuals who are first-degree relatives of one or more Type-2 diabetes patients; genotype, prior history of gestational diabetes, physical inactivity, and race/ethnicity (African Americans, Hispanic/Latino Americans, American Indians, and some Asian Americans and Pacific Islanders are at particularly high risk for developing these diseases).
  • IGT and IFG are themselves risk factors for developing Type-2 diabetes.
  • Polypeptide refers to two or more amino acids joined by one or more peptide bonds.
  • “Pharmaceutically acceptable carrier or excipient” includes, for example, saline, buffered saline, dextrose, water, glycerol, ethanol, lactose, phosphate, mannitol, arginine, trehalose and combinations thereof and further includes agents which enhance the half-life in vivo of GIP, or a biologically active variant thereof, in order to enhance or prolong the biological activity of the peptide or variant.
  • a molecule or chemical moiety may be covalently linked to GIP, or a biologically active variant thereof; or the enhancing agent can be administered simultaneously with GIP, or a biologically active variant thereof.
  • the present invention is directed to methods and kits using GIP or GIP variant to determine whether an individual is susceptible to developing Type-2 diabetes, IGT, or IFG.
  • the present invention includes A method for determining whether an individual is susceptible to developing impaired glucose tolerance (IGT), impaired fasting glucose (IFG), or Type-2 diabetes, comprising: (i) administering to at least one individual at least one polypeptide selected from the group consisting of a gastric inhibitory polypeptide (GIP) and a biologically active GIP variant, or any combination thereof; wherein the polypeptide is optionally combined with a pharmaceutically acceptable carrier or excipient; (ii) assessing the response of the individual to the administration; (Mi) comparing the response to a constant; and (iv) determining from the comparison whether the individual is susceptible to developing IGT, IFG, or Type- 2 diabetes.
  • IGT impaired glucose tolerance
  • IFG impaired fasting glucose
  • Type-2 diabetes comprising: (i) administering to at least one individual at least one polypeptide selected from the group consisting of a gastric inhibitory polypeptide (GIP) and a biologically active GIP variant, or any combination thereof; wherein the polypeptide is optionally combined with
  • the above method can be applied to test any individual, even if the individual is not considered to fall within a risk category for developing Type-2 diabetes, IGT, or IFG.
  • the above method can also be applied more selectively to those who are at risk for developing these diseases, namely individuals exhibiting at least one indicia of susceptibility for developing Type-2 diabetes, IGT, or IFG.
  • the present invention includes applying the above method to, for example, a blood relative of at least one individual who has Type-2 diabetes, IGT, or IFG; an individual who is 45 years of age or older; an individual who is obese; or an individual who already has IGT or IFG (in the context of diagnosing risk for developing Type-2 diabetes).
  • the above method can be used with or without the further step of administering a nutrient.
  • the above method simply involves the administration of GIP or GIP variant in a single dose and then blood is drawn and a response to the GIP or GIP variant is assessed. There is no need to administer a nutrient in this embodiment.
  • the nutrient in a simple and commercially advantageous form, can be in the form of a meal, akin to a glucose tolerance test or can be administered intravenously or otherwise. If administered, the nutrient is preferably administered prior to the assessing step.
  • the preferred nutrient is glucose.
  • other nutrients can be used including carbohydrates, amino acids, lipids, monoglycerides, diglycerides, triglycerides, fatty acids, or any combination thereof.
  • Carbohydrate nutrients useful in the present method include hexoses or pentoses, specific examples of which are glucose, as mentioned above, dextrose, fructose, galactose, xylitol, mannitol, and sorbitol or any combination thereof.
  • Nutrients also include nutrient derivatives, such as pyruvate and lactate, which are carbohydrate derivatives.
  • Nutrients also include intermediates of nutrient metabolic pathways. For example, pyruvic acid is an intermediate of carbohydrate metabolism.
  • the preferred route of administration of the GIP or GIP variant and nutrient is by intravenous infusion, at the same time or at different times.
  • the GIP or GIP variant and nutrient can be administered orally, enterally, parenterally, or otherwise as would understood to the skilled artisan.
  • the preferred method of assessing the response of an individual to GIP or GIP variant alone or with nutrient is to assess plasma levels of insulin, C-peptide, glucose, or levels of insulin secretion, insulin secretion rate, or any combination thereof.
  • insulin response denotes the insulin level in an individual to whom a GIP or GIP variant has been administered.
  • C- peptide response denotes the C-peptide level in an individual to whom a GIP or GIP variant has been administered.
  • glucose response denotes the glucose level in an individual to whom a GIP or GIP variant has been administered. This assessment is made optionally, before, after, or during the administration step.
  • one or more of these levels are assessed before administration and then after administration in order to assess the difference in the levels, which indicates the response to the adn inistration.
  • Methods of assessing these levels, either in the plasma or elsewhere, are known to the skilled artisan. See, for example, Insulin: Andersen, et al., "Enzyme immunoassay of intact insulin in serum and plasma," Clinical Chemistry 39: 578-582 (1993) (insulin levels); Heding, L. G., "Specific and direct radioimmunoassay for human C-peptide in serum,” Diabetologia 11: .
  • insulin secretion rate is assessed. This is commercially advantageous because radioimmuno and ELISA tests for assessing peripheral blood insulin levels are widely available and inexpensive.
  • the plasma levels are assessed after the administration of the GIP or GIP variant, preferably within 5-60 minutes after and more preferably within about 15 minutes after administration.
  • a catheter is placed in the individual and the GIP or GIP variant is administered through the catheter, and then blood is drawn through the same catheter, the catheter being suitable flushed, optionally, in between.
  • the response to the administration is compared to a "constant."
  • the constant is a value which is used to determine whether the response to the administration described above is significantly different from the. response of a normal individual, i.e., an individual who has a normal, non-reduced, GIP response.
  • the constant can be the arithmetic mean of the response of a population of individuals that do not have reduced levels of c-peptide or insulin or increased levels of glucose after administration of GIP or GIP variant and glucose.
  • the constant can of course be more complex in a mathematical sense as would be understood to the skilled artisan.
  • the constant can also be more complex in the sense that it can based on data from particular sub-populations of individuals, where the sub- populations are selected to provide a more accurate control for the test individual or individuals.
  • the constant is predetermined, meaning that the constant has been calculated prior to the administration of the above described method.
  • the "determining step" involves a comparison of the response of the tested individual or individuals to the constant and a determination, based on the comparison, of whether the tested individual or individuals is susceptible to developing IGT, IFG, or Type-2 diabetes.
  • the mathematical difference between the response of the tested individual or individuals and the constant is calculated and this amount is compared to a second constant.
  • the second constant represents a number or range of numbers, wherein, if the above mathematical difference corresponds to the number or falls within the rage of numbers of the second constant, the tested individual or individuals is determined to be susceptible to developing IGT, IFG, or Type-2 diabetes.
  • the second constant is predetermined, meaning that the second constant has been calculated prior to the administration of the above described method.
  • a determining means is used to carry out the determining step.
  • determining nieans include a computer which is programmed to make the above described calculations or a table, chart, or other similar device which can be used to quickly select the appropriate first and second constants or to make the above described calculations.
  • Computer calculations can also be made on an internet site or similar venue, which contains suitable database for making the above calculations.
  • a database is used which is stratified by age, gender, ethnic group, weight, genotype, and other factors, where for each population a constant, including a suitable ranges, can be selected depending on the characteristics of the individual being tested.
  • the present invention further includes a kit for determining whether an individual is susceptible to developing IGT, IFG, or Type-2 diabetes, comprising at least one polypeptide selected from the group consisting of a gastric inhibitory polypeptide (GIP) and a biologically active GIP variant, or any combination thereof: wherein the polypeptide is optionally combined with a pharmaceutically acceptable carrier or excipient, and at least one means for determining whether the individual is susceptible to developing IGT, IFG, or Type-2 diabetes.
  • GIP gastric inhibitory polypeptide
  • kit refers to a one or more containers or one or more packaging materials used to house the contents of the kit.
  • the container or packaging material provides a sterile, contaminant free environment and preferably the kit contains one or more instructions for directing one on how to use the contents of the kit to carry out the above described methods.
  • the kit includes at least one syringe with GIP or GIP variant, or a combination thereof: (1) in powder form, for example lyophilized, to be reconstituted with, for example, saline, or (2) in a liquid form.
  • the kit can either include or not include a nutrient.
  • the kit includes at least one syringe containing one or more nutrients, preferably glucose, (1) in powder form, for example lyophilized, to be reconstituted with, for example, saline, or (2) in a suitable stable liquid form.
  • an I.V. line is installed in the hand or other location of an individual and, if nutrient is to be administered, a glucose or other nutrient syringe is discharged into the IN. line and then a GIP or GIP variant syringe is discharged into the line. After a suitable time interval, blood is drawn from the I.V. line to obtain C-peptide, insulin, or glucose levels. The levels of C-peptide, insulin, or glucose are evaluated as described above to determine whether the individual is susceptible to developing IGT, IFG, or Type-2 diabetes.
  • the kit further includes at least one means for determining whether the individual is susceptible to developing IGT, IFG, or Type-2 diabetes.
  • the means as described above can include a chart or table, or other similar device which can be used to quickly select the appropriate above-described first and second constants or make the above described calculations.
  • the means can also include software or a database for making the calculation or other similar electronic means.
  • the kit can also include a password or other similar code which enables a user to log on to an internet site or other suitable venue as described above to make the calculations described above.
  • Subjects Ten healthy control subjects, ten Type-2 diabetic patients, and 21 first- degree relatives of Type-2 diabetic patients were studied. Subject/patient characteristics are presented in Table 1. The groups were matched for sex, obesity, and age. Non-diabetic participants were subjected to an oral glucose tolerance test (75 g; Boebringer O.G.T., Roche Diagnostics, Mannheim, Germany) with the determination of capillary glucose in the fasting state and 120 min after the ingestion of glucose. 1 subject with diabetes was excluded from the group of relatives. In healthy control subjects, any first- or second-degree relatives with Type-2 diabetes were excluded by history taking.
  • Blood pressure was determined according to the Riva-Rocci method.
  • Five Type-2-diabetic patients had been treated with diet alone, and 5 patients received oral antidiabetic treatment (glibenclamide, 3,5 mg/d, in 1 case, acarbose 150 mg/d in 3 cases, metformin, 1700 mg/d in 1 case). None of the patients had been treated with insulin. In these patients, the usual antidiabetic medication was withdrawn the day before the study.
  • Synthetic GIP was purchased from Poly Peptide Laboratories GmbH, Wolfenb ⁇ ttel, Germany. The lot number (pharmaceutical grade) was: C-0229, net peptide content was 80.3 % . The peptide was dissolved in 0.9 % NaCl/1 % human serum albumin (HSA Behring, salt poor, Marburg, Germany), filtered through 0.2 ⁇ m nitrocellulose filters (Sartorius, G ⁇ ttingen, Germany) and stored frozen at -28 °C as previously described. HPLC profiles (provided by the manufacturer) showed that the preparation was> 99 % pure (single peak coeluting with appropriate standards). Samples were analysed for bacterial growth (standard culture techniques) and for pyrogens (Laboratory Dr. Balfanz, Miinster, Germany). No bacterial contamination was detected. Endotoxin concentrations in samples from the GIP stock solution were 1:61 EU/ml.
  • Glucose was measured using a glucose oxidase method with a Glucose Analyser 2 (Beckman Instruments, Kunststoff, Germany). Insulin was measured using an insulin microparticle enzyme immunoassay (MEIA), IMx Insulin, Abbott Laboratories,
  • C-peptide MTPL EIA C-peptide-antibody-coated microtitre wells
  • IR-GIP was determined as previously described (26), using antiserum R 65 (final dilution 1:150 000) and synthetic human GIP for tracer preparation and as standard. The experimental detection limit was ⁇ 1 pmol/1. Antiserum R 65 binds to the midportion of the GIP molecule. Intra-assay coefficients of variation were » 8% , inter-assay coefficients of variation were ⁇ 6% .
  • IR-Glucagon was measured using porcine antibody 4305 in ethanol- extracted plasma, as previously described (27). The detection limit was ⁇ 1 pmol/1. Intra-assay coefficients of variation were 6.7 %, inter-assay coefficients of variation were 16% .
  • Insulin-resistance and B-cell function were calculated according to the HOMA-model (28).
  • Type-2 diabetic patients had higher fasting plasma glucose and HbA lc concentrations, but lower HDL cholesterol and creatinine concentrations (Table 1). There were no significant differences in any parameter between healthy control subjects and first degree relatives (Table 1):
  • Type-2 diabetic patients were hyperglycemic in the basal state (Fig. 1 A). Steady state glucose concentrations did not differ between the groups (Fig. 1 A). During the infusion of GIP, similar steady-state plasma levels were determined in healthy control subjects, first-degree relatives, and Type-2 diabetic patients, respectively (Fig. 1 B).
  • the insulinotropic response to GIP was significantly lower in Type-2 diabetic patients in comparison to healthy control subjects, no matter whether based on insulin (Fig. 3 A), C-peptide (Fig. 3 B), or the insulin secretion rates (Fig. 3 C).
  • Hyperglycaemia induced a reduction in glucagon concentrations in control subjects and the first-degree-relatives, whereas in Type-2 diabetic patients the values did not change significantly.
  • glucagon concentrations continued to decline in control subjects and first-degree-relatives, but did not change in Type-2 diabetic patients (Fig. 5).
  • GIP has lost part of its insulinotropic effect in at least a subgroup of first-degree relatives of Type-2 diabetic patients (Figs. 2 and 3). This is similar to a well-recognised phenotypic abnormality in Type-2 diabetic patients (7, 24, 25). According to the present study, this reduced insulinotropic effect of GIP precedes any clinically relevant disturbance of glucose tolerance, because the first-degree relatives all had a normal or (in one subject) an impaired oral glucose tolerance.
  • the distribution of insulin secretory responses to the exogenous administration of GIP suggests that approximately 50 % of the first-degree relatives show a normal response, while at least half of them respond very much like Type-2 diabetic patients, i.e. with a markedly reduced insulin secretory response towards GIP (Fig.
  • the present invention therefore comprehends that a reduced insulinotropic response after GIP is an early marker of a predisposition to develop Type-2 diabetes.
  • the present invention further comprehends that the reduced insulinotropic response after GIP also precedes other metabolic disturbances characteristic for Type-2 diabetes like insulin resistance (32, 33), hyperproinsulmemia (34, 35), and diminished B-cell secretory capacity (15, 18), as none of these factors was present in the first-degree-relatives in the present study.
  • a reduced insulinotropic effectiveness of GIP is an early marker that characterises an abnormality of B cell function which might predispose to Type-2 diabetes.
  • the cause of the reduced insulinotropic effectiveness of GIP in Type-2 diabetic patients and in the tested first-degree relatives is not known. It could be a specific defect, for example concerning the level of expression of GIP receptors on pancreatic B cells in Type-2 diabetic patients (14).
  • One possibility is mutations in the GIP receptor leading to an impaired interaction with its ligand, GIP, or a reduced expression of the GIP receptor due to reduced mRNA transcription, translation, or posttranslational modifications affecting its biological activity.
  • GIP receptor coding Polymorphisms in the GIP receptor coding (36) or promoter region in humans, however, have not been found associated with Type-2 diabetes. It is not very likely that other components of the GIP signal transduction pathway are defective, because even in Type-2 diabetic patients, GLP-1 still is very effective in augmenting insulin secretory responses (3, 7, 37, 38). GIP and GLP-1 share most of the components of intracellular signal transduction apart from their receptor molecules, which are different and do not crossreact with the other ligand, respectively (39-45). This also would point to a GIP-specific rather than a general impairment of B cell function in Type-2 diabetic patients and, with all likelihood, also in their first-degree relatives.
  • the present invention further comprehends that the impairment in GIP function found in the present study is one of several aspects of reduced B cell function in more general terms, including a reduced responsiveness to glucose, arginine and possibly other secretagogues (34, 46, 47).
  • a reduced B cell function has also been found in first-degree relatives of Type-2 diabetic patients with different stimuli (18, 20, 48,49).
  • the reduction in measures of B cell secretory parameters relative to normal subjects also in the fasting state and under hyperglycemic conditions found in the present examination could be interpreted in favour of this hypothesis.
  • Another mechanism contemplated by the present invention is that GIP and glucose might act in a synergistic way in stimulating B-cells in the fasting state as well as under hyperglycemic conditions.
  • Insulin resistance is another phenotypic peculiarity of Type-2 diabetic patients, and it has also been recognised in first-degree relatives (16-21, 49). Since it is generally accepted that both secretion defects and a reduced insulin sensitivity have to be present in order to explain all facets of Type-2 diabetes, it was of interest whether our first-degree relatives display features of both insulin resistance and impaired insulin secretion. Using HOMA analysis may have limitations (60), but as far as it can be said, insulin resistance (as determined by the HOMA model) was not a characteristic feature of the same subjects that displayed a reduced insulinotropic effectiveness of exogenous GIP. Therefore, we have, characterised a reduction in B cell secretory function and not the combined occurrence of B cell dysfunction and insulin resistance.
  • GLP-1 Glucagon-like peptide 1
  • GLP-1 an intestinal hormone, signalling nutritional abundance, with an unusual therapeutic potential.
  • TEM 10 229-235.
  • GLP-1/GIP chimeric peptides define the structural requirements for specific Iigand-receptor interaction of GLP-1. Regul Pept 63: 17-22.
  • Pancreatic beta-cells are rendered glucose-competent by the unsulinotropic hormone glucagon-like peptide-l(7-37). Nature 361: 362-365. 51. Krarup, T. 1988. Irnmunoreactive gastric inhibitory polypeptide. Endocrin Reviews 9: 122-133.
  • Exendin (9-39)amide is an antagonist of glucagon-like peptide-1 (7-36)amide in humans. J Clin Invest 101: 1421-1430.

Landscapes

  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Molecular Biology (AREA)
  • Chemical & Material Sciences (AREA)
  • Biomedical Technology (AREA)
  • Urology & Nephrology (AREA)
  • Hematology (AREA)
  • Immunology (AREA)
  • Cell Biology (AREA)
  • Analytical Chemistry (AREA)
  • Biotechnology (AREA)
  • Pathology (AREA)
  • Food Science & Technology (AREA)
  • Medicinal Chemistry (AREA)
  • Physics & Mathematics (AREA)
  • Microbiology (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
  • Investigating Or Analysing Biological Materials (AREA)
  • Peptides Or Proteins (AREA)

Abstract

L'invention concerne des méthodes et des kits permettant de déterminer si un individu est susceptible de développer une intolérance au glucose (IGT), une glycémie à jeun (IFG), ou des diabètes de type 2. Ces méthodes consistent à administrer un polypeptide inhibiteur gastrique ou un variant de polypeptide inhibiteur gastrique et une substance nutritive à un individu, mesurer la réponse de l'individu et déterminer si cet individu est susceptible de développer une intolérance au glucose (IGT), des diabètes de type 2 ou une glycémie à jeun (IFG).
EP01930773A 2000-04-27 2001-04-26 Test de diagnostic de polypeptide inhibiteur gastrique destine a la detection de la sensibilite aux diabetes de type 2, a une intolerance au glucose, ou a une glycemie a jeun Withdrawn EP1356295A2 (fr)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
US55977900A 2000-04-27 2000-04-27
US559779 2000-04-27
PCT/US2001/013378 WO2001081919A2 (fr) 2000-04-27 2001-04-26 Test de diagnostic de polypeptide inhibiteur gastrique destine a la detection de la sensibilite aux diabetes de type 2, a une intolerance au glucose, ou a une glycemie a jeun

Publications (1)

Publication Number Publication Date
EP1356295A2 true EP1356295A2 (fr) 2003-10-29

Family

ID=24234987

Family Applications (1)

Application Number Title Priority Date Filing Date
EP01930773A Withdrawn EP1356295A2 (fr) 2000-04-27 2001-04-26 Test de diagnostic de polypeptide inhibiteur gastrique destine a la detection de la sensibilite aux diabetes de type 2, a une intolerance au glucose, ou a une glycemie a jeun

Country Status (5)

Country Link
EP (1) EP1356295A2 (fr)
KR (1) KR20020097236A (fr)
AU (1) AU2001257278A1 (fr)
CA (1) CA2405900A1 (fr)
WO (1) WO2001081919A2 (fr)

Families Citing this family (28)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20050272652A1 (en) 1999-03-29 2005-12-08 Gault Victor A Peptide analogues of GIP for treatment of diabetes, insulin resistance and obesity
BRPI0606992A2 (pt) * 2005-02-11 2009-07-28 Amylin Pharmaceuticals Inc análogo e polipeptìdeos hìbridos de gip com propriedades selecionáveis
US8263545B2 (en) 2005-02-11 2012-09-11 Amylin Pharmaceuticals, Inc. GIP analog and hybrid polypeptides with selectable properties
EP1959986B1 (fr) 2005-11-07 2014-07-23 Indiana University Research and Technology Corporation Analogues de glucagon a solubilite et a stabilite physiologiques ameliorees
WO2007132291A2 (fr) * 2006-05-15 2007-11-22 Digilab, Inc. Biomarqueurs pour une préforme de diabète de type 2 et méthodes de détection de la présence ou de l'absence d'une préforme du diabète de type 2
EP1857818A1 (fr) * 2006-05-15 2007-11-21 DIGILAB BioVisioN GmbH Diagnostic et usages thérapeutiques des peptides pour les formes de diabète précoces de type 2 et les conditions correspondantes
US8497240B2 (en) 2006-08-17 2013-07-30 Amylin Pharmaceuticals, Llc DPP-IV resistant GIP hybrid polypeptides with selectable properties
CN101578102B (zh) 2007-01-05 2013-07-17 印第安纳大学研究及科技有限公司 在生理性pH缓冲液中显示增强的溶解性的胰高血糖素类似物
EP2111414B1 (fr) 2007-02-15 2014-07-02 Indiana University Research and Technology Corporation Co-agonistes des récepteurs du glucagon/glp-1
EP2217701B9 (fr) 2007-10-30 2015-02-18 Indiana University Research and Technology Corporation Antagonistes du glucagon
MX2010004298A (es) 2007-10-30 2010-05-03 Univ Indiana Res & Tech Corp Compuestos que exhiben actividad antagonista de glucagon y agonista de glp-1.
PA8830501A1 (es) 2008-06-17 2010-07-27 Univ Indiana Res & Tech Corp Co-agonistas del receptor de glucagon/glp-1
EA020326B9 (ru) 2008-06-17 2015-03-31 Индиана Юниверсити Рисерч Энд Текнолоджи Корпорейшн Агонисты смешанного действия на основе глюкозозависимого инсулинотропного пептида для лечения нарушений обмена веществ и ожирения
MX2010012695A (es) 2008-06-17 2011-03-15 Univ Indiana Res & Tech Corp Analogos de glucagon que exhiben solubilidad y estabilidad aumentadas en soluciones reguladoras de ph fisiologico.
RU2550696C2 (ru) 2008-12-19 2015-05-10 Индиана Юниверсити Рисерч Энд Текнолоджи Корпорейшн Основанные на амидах пролекарства пептидов глюкагонового надсемейства
WO2010148089A1 (fr) 2009-06-16 2010-12-23 Indiana University Research And Technology Corporation Composés de glucagon actifs sur le récepteur gip
WO2011075393A2 (fr) 2009-12-18 2011-06-23 Indiana University Research And Technology Corporation Co-agonistes du récepteur du glucagon/glp-i
KR20120123443A (ko) 2010-01-27 2012-11-08 인디애나 유니버시티 리서치 앤드 테크놀로지 코퍼레이션 대사 장애 및 비만 치료용 글루카곤 길항제-ｇｉｐ 항진제 콘쥬게이트
EP2568993A4 (fr) 2010-05-13 2014-02-19 Univ Indiana Res & Tech Corp Peptides de la superfamille du glucagon manifestant une activité de récepteur couplé à une protéine g
WO2011143209A1 (fr) 2010-05-13 2011-11-17 Indiana University Research And Technology Corporation Peptides de la superfamille des glucagons présentant une activité de récepteur nucléaire d'hormone
CN103179979A (zh) 2010-06-24 2013-06-26 印第安纳大学研究及科技有限公司 基于酰胺的胰高血糖素超家族肽前药
MA34885B1 (fr) 2010-12-22 2014-02-01 Indiana Unversity Res And Technology Corp Analogues du glucagon presentant una ctivite de recepteur de gip
RU2014101697A (ru) 2011-06-22 2015-07-27 Индиана Юниверсити Рисерч Энд Текнолоджи Корпорейшн Коагонисты рецепторов глюкагона/glp-1
LT2723367T (lt) 2011-06-22 2017-08-25 Indiana University Research And Technology Corporation Bendri gliukagono/glp-1 receptoriaus agonistai
BR112014007124A2 (pt) 2011-11-17 2017-06-13 Univ Indiana Res & Tech Corp superfamília de peptídeos gluxagon que exibem atividade no receptor glicocorticóide
AR091477A1 (es) 2012-06-21 2015-02-04 Univ Indiana Res & Tech Corp Analogos de glucagon que presentan actividad de receptor de gip
EP3693738A1 (fr) * 2013-09-20 2020-08-12 Astute Medical, Inc. Procédés et compositions pour le diagnostic et le pronostic de l'appendicite et la différenciation des causes de la douleur abdominale
US10317359B2 (en) 2016-01-05 2019-06-11 Ravi Kumar Meruva Differential carbon dioxide sensor

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH04145099A (ja) * 1990-10-05 1992-05-19 Sanwa Kagaku Kenkyusho Co Ltd Ｇｉｐ様活性を有するポリペプチド誘導体及びその用途

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
See references of WO0181919A2 *

Also Published As

Publication number Publication date
KR20020097236A (ko) 2002-12-31
WO2001081919A3 (fr) 2003-04-24
CA2405900A1 (fr) 2001-11-01
WO2001081919A2 (fr) 2001-11-01
AU2001257278A1 (en) 2001-11-07

Similar Documents

Publication Publication Date Title
EP1356295A2 (fr) Test de diagnostic de polypeptide inhibiteur gastrique destine a la detection de la sensibilite aux diabetes de type 2, a une intolerance au glucose, ou a une glycemie a jeun
Meier et al. Reduced insulinotropic effect of gastric inhibitory polypeptide in first-degree relatives of patients with type 2 diabetes
CN1942767A (zh) 对2型糖尿病、葡萄糖耐量低减、空腹血糖异常的易感性测定的肠抑胃肽判断试验
Meier et al. Secretion of incretin hormones and the insulinotropic effect of gastric inhibitory polypeptide in women with a history of gestational diabetes
Rickels et al. High residual C-peptide likely contributes to glycemic control in type 1 diabetes
Knop et al. Reduced incretin effect in type 2 diabetes: cause or consequence of the diabetic state?
Vilsbøll et al. The pathophysiology of diabetes involves a defective amplification of the late-phase insulin response to glucose by glucose-dependent insulinotropic polypeptide—regardless of etiology and phenotype
Knop et al. Impaired incretin effect and fasting hyperglucagonaemia characterizing type 2 diabetic subjects are early signs of dysmetabolism in obesity
Vilsbøll et al. Defective amplification of the late phase insulin response to glucose by GIP in obese Type II diabetic patients
Gavin III et al. Report of the expert committee on the diagnosis and classification of diabetes mellitus
DeFronzo et al. Preservation of β-cell function: the key to diabetes prevention
DeFronzo et al. Fasting hyperglycemia in non-insulin-dependent diabetes mellitus: contributions of excessive hepatic glucose production and impaired tissue glucose uptake
Nauck et al. Secretion of incretin hormones (GIP and GLP-1) and incretin effect after oral glucose in first-degree relatives of patients with type 2 diabetes
Yabe et al. Early phase glucagon and insulin secretory abnormalities, but not incretin secretion, are similarly responsible for hyperglycemia after ingestion of nutrients
Knop et al. The insulinotropic effect of GIP is impaired in patients with chronic pancreatitis and secondary diabetes mellitus as compared to patients with chronic pancreatitis and normal glucose tolerance
US8367418B2 (en) Method and system to provide personalized pharmaceutical compositions and dosages
Calanna et al. Alpha-and beta-cell abnormalities in haemoglobin A1c-defined prediabetes and type 2 diabetes
Svensson et al. A small reduction in glomerular filtration is accompanied by insulin resistance in type I diabetes patients with diabetic nephrophathy
Meier et al. Similar insulin secretory response to a gastric inhibitory polypeptide bolus injection at euglycemia in first-degree relatives of patients with type 2 diabetes and control subjects
Ito et al. Impact of glucagon response on early postprandial glucose excursions irrespective of residual β‐cell function in type 1 diabetes: A cross‐sectional study using a mixed meal tolerance test
Schou et al. Normal secretion and action of the gut incretin hormones glucagon-like peptide-1 and glucose-dependent insulinotropic polypeptide in young men with low birth weight
Schrader et al. Impaired glucose-induced glucagon suppression after partial pancreatectomy
Nandhini et al. Definition, diagnostic criteria, screening, diagnosis, and classification of diabetes and categories of glucose intolerance
Ilag et al. Reduced pancreatic polypeptide response to hypoglycemia and amylin response to arginine in subjects with a mutation in the HNF-4alpha/MODY1 gene.
Safarzad et al. Lower serum zinc level is associated with higher fasting insulin in type 2 diabetes mellitus (T2DM) and relates with disturbed glucagon suppression response in male patients

Legal Events

Date Code Title Description
PUAI Public reference made under article 153(3) epc to a published international application that has entered the european phase

Free format text: ORIGINAL CODE: 0009012

17P Request for examination filed

Effective date: 20021030

AK Designated contracting states

Kind code of ref document: A2

Designated state(s): AT BE CH CY DE DK ES FI FR GB GR IE IT LI LU MC NL PT SE TR

AX Request for extension of the european patent

Extension state: AL LT LV MK RO SI

STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: THE APPLICATION IS DEEMED TO BE WITHDRAWN

18D Application deemed to be withdrawn

Effective date: 20041102