EP0929576A1 - Glp-2 derivate - Google Patents

Glp-2 derivate

Info

Publication number
EP0929576A1
EP0929576A1 EP97938802A EP97938802A EP0929576A1 EP 0929576 A1 EP0929576 A1 EP 0929576A1 EP 97938802 A EP97938802 A EP 97938802A EP 97938802 A EP97938802 A EP 97938802A EP 0929576 A1 EP0929576 A1 EP 0929576A1
Authority
EP
European Patent Office
Prior art keywords
glp
group
derivative according
lys
lipophilic substituent
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Withdrawn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
EP97938802A
Other languages
English (en)
French (fr)
Inventor
Liselotte Bjerre Knudsen
Per Olaf Huusfeldt
Per Franklin Nielsen
Lars Thim
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.)
Novo Nordisk AS
Original Assignee
Novo Nordisk AS
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 Novo Nordisk AS filed Critical Novo Nordisk AS
Publication of EP0929576A1 publication Critical patent/EP0929576A1/de
Withdrawn legal-status Critical Current

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/575—Hormones
    • C07K14/605—Glucagons
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • A61K38/22—Hormones
    • A61K38/26—Glucagons
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • A61K38/22—Hormones
    • A61K38/28—Insulins
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/04—Anorexiants; Antiobesity agents

Definitions

  • the present invention relates to novel derivatives of human glucagon-like peptide-2 (hGLP-2) and analogues thereof and fragments thereof and analogues of such fragments which have a protracted profile of action and to methods of making and using them.
  • hGLP-2 human glucagon-like peptide-2
  • Peptides are widely used in medical practice, and since they can be produced by recombinant DNA technology it can be expected that their importance will increase also in the years to come. When native peptides or analogues thereof are used in therapy it is generally found that they have a high clearance. A high clearance of a therapeutic agent is inconvenient in cases where it is desired to maintain a high blood level thereof over a prolonged period of time since repeated administrations will then be necessary.
  • peptides which have a high clearance are: ACTH, corticotropin-releasing factor, angiotensin, calcitonin, insulin, glucagon, glucagon-like peptide-1 , glucagon-like peptide-2, insulin-like growth factor-1 , insulin-like growth factor-2, gastric inhibitory peptide, growth hormone- releasing factor, pituitary adenylate cyclase activating peptide, secretin, enterogastrin, somatostatin, somatotropin, somatomedin, parathyroid hormone, thrombopoietin, erythropoietin, hypothalamic releasing factors, prolactin, thyroid stimulating hormones, endorphins, enkephalins, vasopressin, oxytocin, opiods and analogues thereof, superoxide dismutase, interferon, asparaginase, arginase, arginine deamina
  • GLP-2 and other preproglucagon fragments are given i.a. by Schmidt et al. (Diabetologia 28 704-707 (1985). Little is known about the physical chemical properties of GLP-2 but GLP-2 is excpected, like GLP-1, to be a highly flexible and unstable molecule. GLP-2 and fragments thereof and analogues of GLP-2 and fragments thereof are potentially useful i.a. in regulation of appetite and in the treatment of small bowel syndrome. However, the high clearance limits the usefulness of these compounds, and thus there still is a need for improvements in this field.
  • Preproglucagon from which GLP-2 originates, is synthesized i.a. in the L-cells in the distal illeum, in the pancreas and in the brain. Processing of preproglucagon to give GLP-1 and GLP-2 occurs mainly in the L-cells.
  • GLP-2 is a 34 amino acid residue peptide.
  • a simple system is used to describe fragments, analogues and derivatives of GLP-2.
  • Lys 20 GLP-2(1-33) designates a fragment of GLP-2 formally derived from GLP-2 by deleting the amino acid residues No. 34 and substituting the naturally occurring amino acid residue in position 20 (Arg) by Lys.
  • Arg ⁇ Lys ⁇ N'-tetradecanoy GLP-IO-S ⁇ designates a derivative of a GLP-2 analogue formally derived from GLP-2 by C-terminal addition of a Lys residue, exchange of the naturally occurring amino acid residue in position 30 (Lys) with an Arg residue and tetradecanoylation of the ⁇ -amino group of the Lys residue in position 35.
  • the present invention relates to derivatives of GLP-2 and analogues thereof.
  • the derivatives according to the invention have interesting pharmacological properties, in particular they have a more protracted profile of action than the parent peptides.
  • GLP-2 designates human GLP-2.
  • an analogue is used to designate a peptide wherein one or more amino acid residues of the parent peptide have been substituted by another amino acid residue and/or wherein one or more amino acid residues of the parent peptide have been deleted and/or wherein one or more amino acid residues have been added to the parent peptide.
  • derivative is used in the present text to designate a peptide in which one or more of the amino acid residues have been chemically modified, e.g. by alkylation, acylation, ester formation or amide formation.
  • a GLP derivative is used in the present text to designate a derivative of GLP-2 or an analogue thereof.
  • the parent peptide from which such a derivative is formally derived is in some places referred to as the "GLP moiety" of the derivative.
  • the present invention relates to a GLP-2 derivative having a lipophilic substituent attached to any one amino acid residue.
  • the present invention relates to a GLP-2 derivative having a lipophilic substituent attached to any one amino acid residue with the proviso that only if the substituent has an ⁇ -carboxylic acid group or is an alkyl group can it be attached to the N-terminal or C-terminal amino acid residue of the parent peptide.
  • the present invention relates to a GLP-2 derivative wherein the lipophilic substituent comprises from 4 to 40 carbon atoms, more preferred from 8 to 25 carbon atoms.
  • the present invention relates to a GLP-2 derivative wherein a lipophilic substituent is attached to an amino acid residue in such a way that a carboxyl group of the lipophilic substituent forms an amide bond with an amino group of the amino acid residue.
  • the present invention relates to a GLP-2 derivative wherein a lipophilic substituent is attached to an amino acid residue in such a way that an amino group of the lipophilic substituent forms an amide bond with a carboxyl group of the amino acid residue.
  • the present invention relates to a GLP-2 derivative wherein a lipophilic substituent is attached to the parent peptide by means of a spacer.
  • the present invention relates to a GLP-2 derivative wherein a lipophilic substituent is attached to the parent peptide by means of a spacer which is an unbranched alkane ⁇ , ⁇ -dicarboxylic acid group having from 1 to 7 methylene groups, preferably two methylene groups which spacer forms a bridge between an amino group of the parent peptide and an amino group of the lipophilic substituent.
  • a spacer which is an unbranched alkane ⁇ , ⁇ -dicarboxylic acid group having from 1 to 7 methylene groups, preferably two methylene groups which spacer forms a bridge between an amino group of the parent peptide and an amino group of the lipophilic substituent.
  • the present invention relates to a GLP-2 derivative wherein a lipophilic substituent is attached to the parent peptide by means of a spacer which is an amino acid residue except Cys, or a dipeptide such as Gly-Lys.
  • a dipeptide such as Gly-Lys is used to designate a dipeptide wherein the C- terminal amino acid residue is Lys, His or Trp, preferably Lys, and wherein the N-terminal amino acid residue is selected from the group comprising Ala, Arg, Asp, Asn, Gly, Glu, Gin, lie, Leu.Val, Phe and Pro.
  • the present invention relates to a GLP-2 derivative wherein a lipophilic substituent is attached to the parent peptide by means of a spacer which is an amino acid residue except Cys, or is a dipeptide such as Gly-Lys and wherein a carboxyl group of the parent peptide forms an amide bond with an amino group of a Lys residue or a dipeptide containing a Lys residue, and the other amino group of the Lys residue or a dipeptide containing a Lys residue forms an amide bond with a carboxyl group of the lipophilic substituent.
  • a spacer which is an amino acid residue except Cys, or is a dipeptide such as Gly-Lys and wherein a carboxyl group of the parent peptide forms an amide bond with an amino group of a Lys residue or a dipeptide containing a Lys residue, and the other amino group of the Lys residue or a dipeptide containing a Lys residue forms an amide bond with a
  • the present invention relates to a GLP-2 derivative wherein a lipophilic substituent is attached to the parent peptide by means of a spacer which is an amino acid residue except Cys, or is a dipeptide such as Gly-Lys and wherein an amino group of the parent peptide forms an amide bond with a carboxylic group of the amino acid or dipeptide spacer, and an amino group of the amino acid or dipeptide spacer forms an amide bond with a carboxyl group of the lipophilic substituent.
  • a spacer which is an amino acid residue except Cys, or is a dipeptide such as Gly-Lys
  • an amino group of the parent peptide forms an amide bond with a carboxylic group of the amino acid or dipeptide spacer
  • an amino group of the amino acid or dipeptide spacer forms an amide bond with a carboxyl group of the lipophilic substituent.
  • the present invention relates to a GLP-2 derivative wherein a lipophilic substituent is attached to the parent peptide by means of a spacer which is an amino acid residue except Cys, or is a dipeptide such as Gly-Lys and wherein a carboxyl group of the parent peptide forms an amide bond with an amino group of the amino acid or dipeptide spacer, and the carboxyl group of the amino acid or dipeptide spacer forms an amide bond with an amino group of the lipophilic substituent.
  • the present invention relates to a GLP-2 derivative wherein a lipophilic substituent is attached to the parent peptide by means of a spacer which is an amino acid residue except Cys, or is a dipeptide such as Gly-Lys, and wherein a carboxyl group of the parent peptide forms an amide bond with an amino group of Asp or Glu, or a dipeptide containing an Asp or Glu residue, and a carboxyl group of the spacer forms an amide bond with an amino group of the lipophilic substituent.
  • a spacer which is an amino acid residue except Cys, or is a dipeptide such as Gly-Lys
  • the present invention relates to a GLP-2 derivative having a lipophilic substituent which comprises a partially or completely hydrogenated cyclopentano- phenathrene skeleton.
  • the present invention relates to a GLP-2 derivative having a lipophilic substituent which is a straight-chain or branched alkyl group.
  • the present invention relates to a GLP-2 derivative having a lipophilic substituent which is the acyl group of a straight-chain or branched fatty acid.
  • the present invention relates to a GLP-2 derivative having a lipophilic substituent which is an acyl group selected from the group comprising CH 3 (CH 2 ) n CO-, wherein n is 4 to 38, preferably CH 3 (CH 2 ) 6 CO-, CH 3 (CH 2 ) 8 CO-, CH 3 (CH 2 ) 10 CO-, CH 3 (CH 2 ) 12 CO-, CH 3 (CH 2 ) 14 CO-, CH 3 (CH 2 ) 16 CO-, CH 3 (CH 2 ) 1 ⁇ CO-, CH 3 (CH 2 ) 20 CO- and CH 3 (CH 2 ) 22 CO-.
  • a lipophilic substituent which is an acyl group selected from the group comprising CH 3 (CH 2 ) n CO-, wherein n is 4 to 38, preferably CH 3 (CH 2 ) 6 CO-, CH 3 (CH 2 ) 8 CO-, CH 3 (CH 2 ) 10 CO-, CH 3 (CH 2 ) 12 CO-, CH 3 (CH 2 ) 14 CO
  • the present invention relates to a GLP-2 derivative having a lipophilic substituent which is an acyl group of a straight-chain or branched alkane ⁇ , ⁇ - dicarboxylic acid.
  • the present invention relates to a GLP-2 derivative having a lipophilic substituent which is an acyl group selected from the group comprising HOOC(CH 2 ) m CO-, wherein m is 4 to 38, preferably HOOC(CH 2 ) 14 CO-, HOOC(CH 2 ) 16 CO-, HOOC(CH 2 ) 18 CO-, HOOCfCH ⁇ CO- and HOOC(CH 2 ) 22 CO-.
  • a lipophilic substituent which is an acyl group selected from the group comprising HOOC(CH 2 ) m CO-, wherein m is 4 to 38, preferably HOOC(CH 2 ) 14 CO-, HOOC(CH 2 ) 16 CO-, HOOC(CH 2 ) 18 CO-, HOOCfCH ⁇ CO- and HOOC(CH 2 ) 22 CO-.
  • the present invention relates to a GLP-2 derivative having a lipophilic substituent which is a group of the formula CH 3 (CH 2 ) p ((CH 2 ) q COOH)CHNH- CO(CH 2 ) 2 CO-, wherein p and q are integers and p+q is an integer of from 8 to 40, preferably from 12 to 35.
  • the present invention relates to a GLP-2 derivative having a lipophilic substituent which is a group of the formula CH 3 (CH 2 ) r CO- NHCH(COOH)(CH 2 ) 2 CO-, wherein r is an integer of from 10 to 24.
  • the present invention relates to a GLP-2 derivative having a lipophilic substituent which is a group of the formula CH 3 (CH 2 ) s CO- NHCH((CH 2 ) 2 COOH)CO-, wherein s is an integer of from 8 to 24.
  • the present invention relates to a GLP-2 derivative having a lipophilic substituent which is a group of the formula COOH(CH 2 ),CO- wherein t is an integer of from 8 to 24.
  • the present invention relates to a GLP-2 derivative having a lipophilic substituent which is a group of the formula -NHCH(COOH)(CH 2 ) 4 NH- CO(CH 2 ) u CH 3 , wherein u is an integer of from 8 to 18.
  • the present invention relates to a GLP-2 derivative having a lipophilic substituent which is a group of the formula -NHCH(COOH)(CH 2 ) 4 NH- COCH((CH 2 ) 2 COOH)NH-CO(CH 2 ) w CH 3 , wherein w is an integer of from 10 to 16.
  • the present invention relates to a GLP-2 derivative having a lipophilic substituent which is a group of the formula -NHCH(COOH)(CH 2 ) 4 NH- CO(CH 2 ) 2 CH(COOH)NH-CO(CH 2 ) !( CH 3 , wherein x is an integer of from 10 to 16.
  • the present invention relates to a GLP-2 derivative having a lipophilic substituent which is a group of the formula -NHCH(COOH)(CH 2 ) 4 NH- CO(CH 2 ) 2 CH(COOH)NHCO(CH 2 ) y CH 3 , wherein y is zero or an integer of from 1 to 22.
  • the present invention relates to a GLP-2 derivative which has one lipophilic substituent.
  • the present invention relates to a GLP-2 derivative which has two lipophilic substituents.
  • the present invention relates to a GLP-2 derivative in which the C-terminal amino acid residue is present in the form of the amide.
  • the present invention relates to a GLP-2 derivative having a lipophilic substituent which can be negatively charged.
  • the present invention relates to a GLP-2 derivative the parent peptide of which is selected from the group comprising GLP-2(1-35) or an analogue thereof.
  • the present invention relates to a GLP-2 derivative derived from a GLP-2 fragment selected from the group comprising GLP-2(1-30); GLP-2(1- 31 ); GLP-2(1-32); GLP-2(1-33); GLP-2(1-34) and GLP-2(1-35).
  • the present invention relates to a GLP-2 derivative wherein the designation analogue comprises derivatives wherein a total of up to ten amino acid residues have been exchanged with any ⁇ -amino acid residue.
  • the present invention relates to a derivative of a GLP-2 analogue wherein the designation analogue implies that the parent peptide is human GLP-2 wherein a total of up to six, more preferred up to three, amino acid residues have been added, deleted or substituted with other amino acid residues which can be coded for by the genetic code.
  • the present invention relates to a GLP-2 derivative wherein the parent peptide is selected from the group comprising Lys 20 GLP-2(1-33) and Lys 20 Arg 30 GLP-2(1-33).
  • the present invention relates to a GLP-2 derivative wherein the parent peptide is Arg 30 Lys 3 GLP-2(1-34).
  • the present invention relates to a GLP-2 derivative wherein the parent peptide is selected from the group comprising Arg 30 Lys 35 GLP-2(1-35); Arg 30 ' 35 Lys 20 GLP-2(1-35) and Arg 35 GLP-2(1-35).
  • the present invention relates to a GLP-2 derivative which is selected from the group comprising
  • Lys 20 (N ⁇ -tetradecanoyl)GLP-2(1-33);
  • Lys 20 (N c -tetradecanoyl)Arg 30 GLP-2(1-33);
  • Lys 20 (N ⁇ -( ⁇ -carboxynonadecanoyl))GLP-2(1-33);
  • Lys 20 (N e -( ⁇ -carboxynonadecanoyl))Arg 30 GLP-2(1-33); Arg 30 Lys 35 (N ⁇ -( ⁇ -carboxynonadecanoyl))GLP-2(1-35);
  • the present invention relates to a pharmaceutical composition
  • a pharmaceutical composition comprising a GLP-2 derivative and a pharmaceutically acceptable vehicle or carrier.
  • the present invention relates to the use of a GLP-2 derivative according to the invention for the preparation of a medicament which has a more protracted action than the parent peptide. In a further preferred embodiment, the present invention relates to the use of a GLP-2 derivative according to the invention for the preparation of a medicament with protracted effect for the treatment of obesity.
  • the present invention relates to the use of a GLP-2 derivative according to the invention for the preparation of a medicament with protracted effect for the treatment of small bowel syndrome.
  • the lipophilic substituent attached to the GLP-2 moiety preferably comprises 4-40 carbon atoms, in particular 8-25 carbon atoms.
  • the lipophilic substituent may be attached to an amino group of the GLP-2 moiety by means of a carboxyl group of the lipophilic substituent which forms an amide bond with an amino group of the amino acid to which it is attached.
  • the lipophilic substituent may be attached to said amino acid in such a way that an amino group of the lipophilic substituent forms an amide bond with a carboxyl group of the amino acid.
  • the lipophililic substituent may be linked to the GLP-2 moiety via an ester bond.
  • the ester can be formed either by reaction between a carboxyl group of the GLP-2 moiety and a hydroxyl group of the substituent-to-be or by reaction between a hydroxyl group of the GLP-2 moiety and a carboxyl group of the substituent-to-be.
  • the lipophilic substituent can be an alkyl group which is introduced into a primary amino group of the GLP-2 moiety.
  • the lipophilic substituent is attached to the GLP-2 moiety by means of a spacer in such a way that a carboxyl group of the spacer forms an amide bond with an amino group of the GLP-2 moiety.
  • suitable spacers are succinic acid, Lys, Glu or Asp, or a dipeptide such as Gly-Lys.
  • the spacer is succinic acid, one carboxyl group thereof may form an amide bond with an amino group of the amino acid residue, and the other carboxyl group thereof may form an amide bond with an amino group of the lipophilic substituent.
  • the spacer is Lys, Glu or Asp
  • the carboxyl group thereof may form an amide bond with an amino group of the amino acid residue
  • the amino group thereof may form an amide bond with a carboxyl group of the lipophilic substituent.
  • a further spacer may in some instances be inserted between the ⁇ -amino group of Lys and the lipophilic substituent.
  • such a further spacer is succinic acid which forms an amide bond with the ⁇ - amino group of Lys and with an amino group present in the lipophilic substituent.
  • such a further spacer is Glu or Asp which forms an amide bond with the ⁇ -amino group of Lys and another amide bond with a carboxyl group present in the lipophilic substituent, that is, the lipophilic substituent is a N c -acylated lysine residue.
  • the lipophilic substituent has a group which can be negatively charged.
  • One preferred group which can be negatively charged is a carboxylic acid group.
  • the parent peptide can be produced by a method which comprises culturing a host cell containing a DNA sequence encoding the peptide and capable of expressing the peptide in a suitable nutrient medium under conditions permitting the expression of the peptide, after which the resulting peptide is recovered from the culture.
  • the medium used to culture the cells may be any conventional medium suitable for growing the host cells, such as minimal or complex media containing appropriate supplements. Suitable media are available from commercial suppliers or may be prepared according to published recipes (e.g. in catalogues of the American Type Culture Collection).
  • the peptide produced by the cells may then be recovered from the culture medium by conventional procedures including separating the host cells from the medium by centrifugation or filtration, precipitating the proteinaceous components of the supernatant or filtrate by means of a salt, e.g. ammonium sulphate, purification by a variety of chromatographic procedures, e.g. ion exchange chromatography, gelfiltration chromatography, affinity chromatography, or the like, dependent on the type of peptide in question.
  • a salt e.g. ammonium sulphate
  • the DNA sequence encoding the parent peptide may suitably be of genomic or cDNA origin, for instance obtained by preparing a genomic or cDNA library and screening for DNA sequences coding for all or part of the peptide by hybridisation using synthetic oligonucleotide probes in accordance with standard techniques (see, for example, Sambrook, J, Fritsch, EF and Maniatis, T, Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory Press, New York, 1989).
  • the DNA sequence encoding the peptide may also be prepared synthetically by established standard methods, e.g.
  • the DNA sequence may also be prepared by polymerase chain reaction using specific primers, for instance as described in US 4,683,202 or Saiki et al., Science 239 (1988), 487 - 491.
  • the DNA sequence may be inserted into any vector which may conveniently be subjected to recombinant DNA procedures, and the choice of vector will often depend on the host cell into which it is to be introduced.
  • the vector may be an autonomously replicating vector, i.e. a vector which exists as an extrachromosomal entity, the replication of which is independent of chromosomal replication, e.g. a plasmid.
  • the vector may be one which, when introduced into a host cell, is integrated into the host cell genome and replicated together with the chromosome(s) into which it has been integrated.
  • the vector is preferably an expression vector in which the DNA sequence encoding the peptide is operably linked to additional segments required for transcription of the DNA, such as a promoter.
  • the promoter may be any DNA sequence which shows transcriptional activity in the host cell of choice and may be derived from genes encoding proteins either homologous or heterologous to the host cell. Examples of suitable promoters for directing the transcription of the DNA encoding the peptide of the invention in a variety of host cells are well known in the art, cf. for instance Sambrook et al., supra.
  • the DNA sequence encoding the peptide may also, if necessary, be operably connected to a suitable terminator, polyadenylation signals, transcriptional enhancer sequences, and translational enhancer sequences.
  • the recombinant vector of the invention may further comprise a DNA sequence enabling the vector to replicate in the host cell in question.
  • the vector may also comprise a selectable marker, e.g. a gene the product of which complements a defect in the host cell or one which confers resistance to a drug, e.g. ampicillin, kanamycin, tetracyclin, chloramphenicol, neomycin, hygromycin or methotrexate.
  • a secretory signal sequence also known as a leader sequence, prepro sequence or pre sequence
  • the secretory signal sequence is joined to the DNA sequence encoding the peptide in the correct reading frame. Secretory signal sequences are commonly positioned 5' to the DNA sequence encoding the peptide.
  • the secretory signal sequence may be that normally associated with the peptide or may be from a gene encoding another secreted protein.
  • the host cell into which the DNA sequence or the recombinant vector is introduced may be any cell which is capable of producing the present peptide and includes bacteria, yeast, fungi and higher eukaryotic cells.
  • suitable host cells well known and used in the art are, without limitation, E. coli, Saccharomyces cerevisiae, or mammalian BHK or CHO cell lines.
  • the GLP-2 derivatives of the invention can be prepared by introducing the lipophilic substituent into the parent GLP-2 or GLP-2 analogue using methods known per se, see for example WO 95/07931, the contents of which is hereby incorporated in its entirety by reference.
  • N E -acylation of a Lys residue can be carried out by using an activated amide of the acyl group to be introduced as the acylating agent, e.g. the amide with benzotriazole.
  • the acylation is carried out in a polar solvent in the presence of a base.
  • compositions containing a GLP-2 derivative according to the present invention may be administered parenterally to patients in need of such a treatment.
  • Parenteral administration may be performed by subcutaneous, intramuscular or intravenous injection by means of a syringe, optionally a pen-like syringe.
  • parenteral administration can be performed by means of an infusion pump.
  • a further option is a composition which may be a powder or a liquid for the administration of the GLP-2 derivative in the form of a nasal or pulmonal spray.
  • the GLP-2 derivatives of the invention can also be administered transdermally, e.g. from a patch, optionally a iontophoretic patch, or transmucosally, e.g. bucally.
  • compositions containing a GLP-2 derivative of the present invention may be prepared by conventional techniques, e.g. as described in Remington's Pharmaceutical Sciences. 1985 or in Remington: The Science and Practice of Pharmacy, 19 th edition, 1995.
  • the injectable compositions of the GLP-2 derivative of the invention can be prepared using the conventional techniques of the pharmaceutical industry which involves dissolving and mixing the ingredients as appropriate to give the desired end product.
  • the GLP-2 derivative is dissolved in an amount of water which is somewhat less than the final volume of the composition to be prepared.
  • An isotonic agent, a preservative and a buffer is added as required and the pH value of the solution is adjusted - if necessary - using an acid, e.g. hydrochloric acid, or a base, e.g. aqueous sodium hydroxide as needed.
  • the volume of the solution is adjusted with water to give the desired concentration of the ingredients.
  • isotonic agents sodium chloride, mannitol and glycerol.
  • preservatives examples include phenol, m-cresol, methyl p-hydroxybenzoate and benzyl alcohol.
  • Suitable buffers are sodium acetate and sodium phosphate.
  • solutions containing a GLP-2 derivative according to the present invention may also contain a surfactant in order to improve the solubility and/or the stability of the derivative.
  • a composition for nasal administration of GLP-2 may, for example, be prepared as described in European Patent No. 272097 (to Novo Nordisk A S) or in WO 93/18785.
  • the GLP-2 derivatives of this invention can be used in the treatment of various diseases.
  • the particular GLP-2 derivative to be used and the optimal dose level for any patient will depend on the disease to be treated and on a variety of factors including the efficacy of the specific peptide derivative employed, the age, body weight, physical activity, and diet of the patient, on a possible combination with other drugs, and on the severity of the case. It is recommended that the dosage of the GLP-2 derivative of this invention be determined for each individual patient by those skilled in the art in a similar way as for known parent peptides.
  • NMP N-Methyl-2-pyrrolidone.
  • EDPA N-Ethyl-N,N-diisopropylamine.
  • TFA Trifluoroacetic acid.
  • Myr-ONSu Tetradecanoic acid 2,5-dioxopyrrolidin-1-yl ester. Abbreviations:
  • PDMS Plasma Desorption Mass Spectrometry
  • HPLC High Performance Liquid Chromatography amu: atomic mass units
  • the reaction was quenched by the addition of a solution of glycine (4.5 mg, 4.5 ⁇ mol) in 50% aqueous ethanol (451 ⁇ l).
  • the reaction mixture was purified by column chromatography using a cyanopropyl column (Zorbax 300SB-CN) and a standard acetonitrile/TFA system. The column was heated to 65°C and the acetonitrile gradient was 0-100% in 60 minutes. The title compound ( 5.0 mg, 47 %) was isolated from the eluate.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • General Health & Medical Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Organic Chemistry (AREA)
  • Veterinary Medicine (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Public Health (AREA)
  • Engineering & Computer Science (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Animal Behavior & Ethology (AREA)
  • Endocrinology (AREA)
  • Zoology (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Diabetes (AREA)
  • General Chemical & Material Sciences (AREA)
  • Epidemiology (AREA)
  • Immunology (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Hematology (AREA)
  • Biochemistry (AREA)
  • Biophysics (AREA)
  • Genetics & Genomics (AREA)
  • Molecular Biology (AREA)
  • Child & Adolescent Psychology (AREA)
  • Toxicology (AREA)
  • Obesity (AREA)
  • Peptides Or Proteins (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)
EP97938802A 1996-08-30 1997-09-01 Glp-2 derivate Withdrawn EP0929576A1 (de)

Applications Claiming Priority (5)

Application Number Priority Date Filing Date Title
DK93196 1996-08-30
DK93196 1996-08-30
DK125996 1996-11-08
DK125996 1996-11-08
PCT/DK1997/000360 WO1998008872A1 (en) 1996-08-30 1997-09-01 Glp-2 derivatives

Publications (1)

Publication Number Publication Date
EP0929576A1 true EP0929576A1 (de) 1999-07-21

Family

ID=26064922

Family Applications (1)

Application Number Title Priority Date Filing Date
EP97938802A Withdrawn EP0929576A1 (de) 1996-08-30 1997-09-01 Glp-2 derivate

Country Status (4)

Country Link
EP (1) EP0929576A1 (de)
JP (1) JP2000517308A (de)
AU (1) AU4112497A (de)
WO (1) WO1998008872A1 (de)

Families Citing this family (39)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
UA65549C2 (uk) 1996-11-05 2004-04-15 Елі Ліллі Енд Компані Спосіб регулювання ожиріння шляхом периферійного введення аналогів та похідних glp-1 (варіанти) та фармацевтична композиція
AU2712899A (en) * 1998-02-27 1999-09-15 Novo Nordisk A/S Glp-2 derivatives with helix-content exceeding 25 percent, forming partially structured micellar-like aggregates
US6444788B1 (en) 1999-03-15 2002-09-03 Novo Nordisk A/S Ion exchange chromatography of GLP-1, analogs and derivatives thereof
WO2000055203A1 (en) 1999-03-15 2000-09-21 Novo Nordisk A/S Ion exchange chromatographic separation of glp-1 and related peptides
US6451987B1 (en) 1999-03-15 2002-09-17 Novo Nordisk A/S Ion exchange chromatography of proteins and peptides
EP1956000B1 (de) 1999-03-17 2016-10-05 Novo Nordisk A/S Acylierungsmittel zur acylierung von peptiden
US6451974B1 (en) 1999-03-17 2002-09-17 Novo Nordisk A/S Method of acylating peptides and novel acylating agents
US7371721B2 (en) 2000-09-18 2008-05-13 Sanos Bioscience A/S Use of GLP-2 and related compounds for the treatment, prevention, diagnosis, and prognosis of bone-related disorders and calcium homeostasis related syndromes
ES2310192T3 (es) 2000-09-18 2009-01-01 Sanos Bioscience A/S Uso de peptidos glp-2.
WO2002040537A2 (en) * 2000-11-16 2002-05-23 Novo Nordisk A/S Analogues and derivatives of gastrin releasing peptide (grp)
CN1491233A (zh) 2001-02-02 2004-04-21 康久化学公司 长效生长激素释放因子衍生物
ATE396202T1 (de) * 2001-02-16 2008-06-15 Conjuchem Biotechnologies Inc Lang wirkendes glucagon-ähnliches peptid-2 für die behandlung von gastrointestinalen krankheiten und störungen
US7595172B2 (en) 2001-07-24 2009-09-29 Novo Nordisk A/S Method for making acylated polypeptides
US20030082671A1 (en) 2001-07-24 2003-05-01 Thomas Hoeg-Jensen Method for making acylated polypeptides
CA2500123A1 (en) * 2002-09-25 2004-04-08 Novo Nordisk A/S Method for producing acylated peptides
US7273921B2 (en) 2002-09-25 2007-09-25 Novo Nordisk A/S Method for producing acylated peptides
WO2004035624A2 (en) * 2002-10-14 2004-04-29 Novo Nordisk A/S Glucagon - like peptide - 2 variants
DE602004031927D1 (de) 2003-02-04 2011-05-05 Novo Nordisk As Injektionsvorrichtung mit drehbarer dosiseinstellungsvorrichtung
WO2004085471A2 (en) * 2003-03-24 2004-10-07 Novo Nordisk A/S Glp-2 derivatives
WO2004089985A1 (en) 2003-04-11 2004-10-21 Novo Nordisk A/S Stable pharmaceutical compositions
EP1664108B1 (de) 2003-08-21 2009-10-14 Novo Nordisk A/S Trennung von polypeptiden mit einer racemisierten aminosäure
JP2007537981A (ja) * 2003-09-19 2007-12-27 ノボ　ノルディスク　アクティーゼルスカブ 新規の血漿タンパク質親和性タグ
KR101241862B1 (ko) * 2003-09-19 2013-03-13 노보 노르디스크 에이/에스 신규 ｇｌｐ-1 유도체
EP1684793B1 (de) * 2003-11-13 2011-09-21 Novo Nordisk A/S Pharmazeutische zusammensetzung umfassend eine insulinotrope glp-1(7-37) analoge, asp(b28)-insulin, und eine oberflächenaktive verbindung
US20060287221A1 (en) 2003-11-13 2006-12-21 Novo Nordisk A/S Soluble pharmaceutical compositions for parenteral administration comprising a GLP-1 peptide and an insulin peptide of short time action for treatment of diabetes and bulimia
EP2033662B1 (de) 2004-01-21 2012-10-17 Novo Nordisk Health Care AG Transglutaminase vermittelte konjugation von peptiden.
WO2005082404A2 (en) * 2004-02-27 2005-09-09 Novo Nordisk A/S Glp-2 derivatives modified by lipophilic substituents
JP5755398B2 (ja) 2005-03-18 2015-07-29 ノヴォ ノルディスク アー／エス 伸長されたｇｌｐ−１化合物
TWI372629B (en) 2005-03-18 2012-09-21 Novo Nordisk As Acylated glp-1 compounds
MX2008009125A (es) 2006-01-18 2008-10-23 Qps Llc Composiciones farmaceuticas con estabilidad mejorada.
EP2054073B1 (de) * 2006-07-11 2014-11-26 Foresee Pharmaceuticals, Inc. Pharmazeutische zusammensetzungen für die verzögerte freisetzung von peptiden
WO2008025856A2 (en) 2006-09-01 2008-03-06 Novo Nordisk Health Care Ag Modified glycoproteins
KR20110039348A (ko) 2008-08-06 2011-04-15 노보 노르디스크 헬스 케어 악티엔게젤샤프트 연장된 생체내 효능을 가지는 콘쥬게이트된 단백질
US8637647B2 (en) 2008-09-12 2014-01-28 Novo Nordisk A/S Method of acylating a peptide or protein
JP5816097B2 (ja) 2009-01-22 2015-11-18 ノヴォ・ノルディスク・ヘルス・ケア・アーゲー 安定な成長ホルモン化合物
JP6086528B2 (ja) 2009-08-06 2017-03-01 ノヴォ・ノルディスク・ヘルス・ケア・アーゲー 長期のインビボ有効性を有する成長ホルモン
CN118767117A (zh) 2010-01-22 2024-10-15 诺沃—诺迪斯克保健股份有限公司 体内功效延长的生长激素
JP5980689B2 (ja) 2010-01-22 2016-08-31 ノヴォ・ノルディスク・ヘルス・ケア・アーゲー 安定な成長ホルモン化合物
US11045523B2 (en) 2013-04-05 2021-06-29 Novo Nordisk Healthcare Ag Formulation of growth hormone albumin-binder conjugate

Family Cites Families (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
ATE164852T1 (de) * 1990-01-24 1998-04-15 Douglas I Buckley Glp-1-analoga verwendbar in der diabetesbehandlung
US5512549A (en) * 1994-10-18 1996-04-30 Eli Lilly And Company Glucagon-like insulinotropic peptide analogs, compositions, and methods of use
US5990077A (en) * 1995-04-14 1999-11-23 1149336 Ontario Inc. Glucagon-like peptide-2 and its therapeutic use

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
See references of WO9808872A1 *

Also Published As

Publication number Publication date
AU4112497A (en) 1998-03-19
WO1998008872A1 (en) 1998-03-05
JP2000517308A (ja) 2000-12-26

Similar Documents

Publication Publication Date Title
WO1998008872A1 (en) Glp-2 derivatives
EP1060192A2 (de) Glp-2 derivate mit einem helixgehalt über 25%, die partiell strukturierte mizellenartige aggregate bilden
EP1062240B1 (de) N-terminal veränderte glp-1 abkömmlinge
AU712096B2 (en) Lipophilic peptide hormone derivatives
EP3157949B1 (de) Exendin-4-derivate als selektive glucagonrezeptoragonisten
US9409966B2 (en) Glucagon-like peptide-1 derivatives and their pharmaceutical use
EP1664108B1 (de) Trennung von polypeptiden mit einer racemisierten aminosäure
US20100261637A1 (en) Peptides derivatized with a-b-c-d- and their therapeutical use
WO1999043705A1 (en) N-terminally truncated glp-1 derivatives
KR20060135661A (ko) 신규 ｇｌｐ-1 화합물
US20020025933A1 (en) GLP-2 derivatives
CN109248323B (zh) 酰化的glp-1衍生物
WO1998045329A1 (en) Natriuretic peptide derivatives
EP3426678B1 (de) Konjugate von inselzellenneogenesepeptiden und analoga sowie verfahren dafür
US20020165148A1 (en) Analogues and derivatives of gastrin releasing peptide (GRP)

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: 19990330

AK Designated contracting states

Kind code of ref document: A1

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

AX Request for extension of the european patent

Free format text: AL PAYMENT 19990330;LT PAYMENT 19990330;LV PAYMENT 19990330;RO PAYMENT 19990330;SI PAYMENT 19990330

17Q First examination report despatched

Effective date: 20030722

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: 20080624