PHARMACOLOGICALLY ACTIVE PEPTIDE CONJUGATES HAVING A REDUCED TENDENCY TOWARDS ENZYMATIC HYDROLYSIS

• US 20110312878A1
• Filed: 02/15/2011
• Published: 12/22/2011
• Est. Priority Date: 03/09/1998
• Status: Abandoned Application

• Current Assignee: Zealand Pharma A/S
• Original Assignee: Zealand Pharma A/S
• Inventors: Larsen, Bjarne Due
• Application Number:
  US13/027,475
• Publication Number:
  US 20110312878A1
• Time in Patent Office: Days
• US Class Current: 514/3.8
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: A61K 47/64 Drug-peptide, drug-protein or drug-polyamino acid conjugates, i.e. the modifying agent being a peptide, protein or polyamino acid which is covalently bonded or complexed to a therapeutically active agent peptidic linkers A61K47/65
• CPC Class Codes: A61K 47/65 Peptidic linkers, binders or spacers, e.g. peptidic enzyme-labile linkers
• CPC Class Codes: A61K 47/67 Enzyme prodrug therapy, e.g. gene directed enzyme drug therapy [GDEPT] or VDEPT
• CPC Class Codes: A61P 1/14 Prodigestives, e.g. acids, enzymes, appetite stimulants, antidyspeptics, tonics, antiflatulents
• CPC Class Codes: A61P 13/00 Drugs for disorders of the urinary system
• CPC Class Codes: A61P 13/02 of urine or of the urinary tract, e.g. urine acidifiers
• CPC Class Codes: A61P 15/18 Feminine contraceptives
• CPC Class Codes: A61P 19/08 for bone diseases, e.g. rachitism, Paget's disease
• CPC Class Codes: A61P 19/10 for osteoporosis
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• CPC Class Codes: A61P 25/04 Centrally acting analgesics, e.g. opioids
• CPC Class Codes: A61P 25/18 Antipsychotics, i.e. neuroleptics; Drugs for mania or schizophrenia
• CPC Class Codes: A61P 25/20 Hypnotics; Sedatives
• CPC Class Codes: A61P 25/24 Antidepressants
• CPC Class Codes: A61P 25/28 for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
• CPC Class Codes: A61P 29/00 Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
• CPC Class Codes: A61P 3/00 Drugs for disorders of the metabolism of the blood or the extracellular fluid A61P7/00
• CPC Class Codes: A61P 3/04 Anorexiants; Antiobesity agents
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: A61P 31/18 : for HIV
• CPC Class Codes: A61P 35/00 : Antineoplastic agents
• CPC Class Codes: A61P 43/00 : Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: A61P 5/00 : Drugs for disorders of the endocrine system
• CPC Class Codes: A61P 5/02 : of the hypothalamic hormones, e.g. TRH, GnRH, CRH, GRH, somatostatin
• CPC Class Codes: A61P 5/06 : of the anterior pituitary hormones, e.g. TSH, ACTH, FSH, LH, PRL, GH
• CPC Class Codes: A61P 5/18 : of the parathyroid hormones
• CPC Class Codes: A61P 5/24 : of the sex hormones
• CPC Class Codes: A61P 7/06 : Antianaemics
• CPC Class Codes: A61P 7/12 : Antidiuretics, e.g. drugs for diabetes insipidus ADH A61P5/10
• CPC Class Codes: A61P 9/12 : Antihypertensives
• CPC Class Codes: B82Y 5/00 : Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery
• CPC Class Codes: C07K 1/1075 : by covalent attachment of amino acids or peptide residues
• CPC Class Codes: C07K 14/575 : Hormones derived from pro-opiomelanocortin, pro-enkephalin or pro-dynorphin C07K14/665, e.g. corticotropin C07K14/695
• CPC Class Codes: C07K 14/57545 : Neuropeptide Y
• CPC Class Codes: C07K 14/665 : derived from pro-opiomelanocortin, pro-enkephalin or pro-dynorphin
• CPC Class Codes: C07K 14/675 : Beta-endorphins
• CPC Class Codes: C07K 14/68 : Melanocyte-stimulating hormone [MSH]
• CPC Class Codes: C07K 14/685 : Alpha-melanotropin
• CPC Class Codes: C07K 14/695 : Corticotropin [ACTH]
• CPC Class Codes: C07K 2319/31 : fusions, other than Fc, for prolonged plasma life, e.g. albumin

Abstract

The invention is directed to a pharmacologically active peptide conjugate having a reduced tendency towards enzymatic cleavage comprising a pharmacologically active peptide sequence (X) and a stabilising peptide sequence (Z) of 4-20 amino acid residues covalently bound to X.

Claims

1. A pharmacologically active peptide conjugate having a reduced tendency towards enzymatic cleavage comprising X and Z,

wherein X is a pharmacologically active peptide sequence selected from the group consisting of endorphins, adrenocorticotropic hormone (ACTH), melanotan II, melanocyte stimulating hormone (MSH), alpha-MSH, and any modified or truncated analogues thereof, and

wherein Z is a stabilizing peptide sequence of 4-15 amino acid units covalently bound to X via a peptide bond to the C-terminal carbonyl function of X and/or to the N-terminal nitrogen atom of X, and wherein Z consists of residues selected from Glu, Lys, and Met, and

wherein the ratio between the half-life of said peptide conjugate and the half-life of the corresponding pharmacologically active peptide sequence X, when treated with carboxypeptidase A or leucine aminopeptidase in about 50 mM phosphate buffer solution at about pH 7.4 at about 37°

C. or in serum or plasma is at least about 2, or wherein said peptide conjugate has a half-life in human or mice serum or plasma at 37°

C. of at least about 10 minutes.

2. The peptide conjugate according to claim 1, wherein Z is covalently bound to X via a peptide bond to the N-terminal nitrogen atom of X.

3. The peptide conjugate according to claim 1, wherein X is alpha-MSH or a modified or truncated analogue thereof.

4. The peptide conjugate according to claim 1, wherein X comprises at least 10 amino acid residues.

5. The peptide conjugate according to claim 3, wherein X is alpha-MSH.

6. The peptide conjugate according to claim 3, wherein X is a modified or truncated analogue of alpha-MSH.

7. The peptide conjugate according to claim 1, wherein X is a modified or truncated analogue in which the amino acid sequence of a pharmacologically active peptide sequence selected from the group consisting of endorphins, adrenocorticotropic hormone (ACTH), melanotan II, melanocyte stimulating hormone (MSH), alpha-MSH, has been modified by changing and/or deleting one or more amino acid residues.

8. The peptide conjugate according to claim 7, wherein X has been modified by altering the side chain stereochemistry and/or backbone in said one amino acid residue.

9. The peptide conjugate according to claim 7 wherein a peptide bond has been changed into a reduced form.

10. The peptide conjugate according to claim 1, wherein Z consists of 4-10 amino acid units.

11. The peptide conjugate according to claim 1, wherein Z consists of 4-7 amino acid units.

12. The peptide conjugate according to claim 1, wherein Z consists of 6 amino acid units.

13. The peptide conjugate according to claim 12, wherein Z is Lys₆.

14. The peptide conjugate according to claim 1, wherein the amino acid units in Z are Lys.

15. The peptide conjugate according to claim 14, wherein Z is selected from the group consisting of Lys₄, Lys₅, and Lys₆.

16. A method for the preparation of a pharmacologically active peptide conjugate (Z-X) as defined in claim 2, comprising the steps of:

a) coupling an N-α

-protected amino acid, or an N-α

-protected dipeptide to a solid support material (SSM), thereby forming an immobilized N-α

-protected amino acid,

b) removing the N-α

-protecting group, thereby forming an immobilized amino acid or peptide fragment having an unprotected N-terminal end,

c) coupling an additional N-α

-protected amino acid in the carboxyl activated form, or an additional N-α

-protected dipeptide in the C-terminal activated form to the N-terminal end of the immobilized amino acid or peptide fragment, and repeating the removal/coupling step procedure in step b) and c) until the desired peptide sequence X is obtained,

d) coupling an additional N-α

-protected amino acid in the carboxyl activated form, or an additional N-α

-protected dipeptide in the C-terminal activated form to the N-terminal end of the immobilized peptide fragment, and repeating the removal/coupling step procedure in step b) and d) until the desired peptide sequence Z is obtained, and then

e) cleaving off the peptide conjugate from the solid support material.

17. A method for producing a peptide conjugate according to claim 1, comprising

a) introducing a nucleic acid sequence encoding said conjugate into a host cell;

b) culturing said host cell and

c) isolating said conjugate from the culture.

18. A method for producing a peptide conjugate according to claim 1, comprising

a) culturing a recombinant host cell comprising a nucleic acid sequence encoding said conjugate under conditions permitting the production of said conjugate;

and

b) isolating said conjugate from the culture.

19. A composition comprising a pharmacologically active peptide conjugate as defined in claim 1 and a pharmaceutical acceptable carrier.

20. The composition of claim 19, wherein said composition is formulated for oral, subcutaneous, parenteral, intramuscular, rectal, epidural, intratracheal, intranasal, vaginal, buccal, ocular, direct brain, pulmonary or topical administration.

21. A method for treating a disease or disorder, wherein the peptide sequence X, when not bound to Z, is able to interact with a receptor or a receptor system involved in the condition or disorder in question, and where the interaction between X, when not bound to Z, and the receptor or receptor system has a therapeutic or prophylactic effect on the condition or disorder, said method comprising administering to a patient in need thereof a peptide conjugate as defined in claim 1.

22. A method for treating a disease or disorder selected from the group consisting of HIV infection, cancer, diabetes, incontinence, hypertension, amnesias, Alzheimer'"'"'s disease, fever, depression, sex hormone regulation, eating, schizophrenia, osteoporosis, and insomnia, said method comprising administering to a patient in need thereof a peptide conjugate as defined in claim 1.