EP0707490A1 - Antagonistes et agonistes du neuropeptide y - Google Patents

Antagonistes et agonistes du neuropeptide y

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Publication number
EP0707490A1
EP0707490A1 EP94920757A EP94920757A EP0707490A1 EP 0707490 A1 EP0707490 A1 EP 0707490A1 EP 94920757 A EP94920757 A EP 94920757A EP 94920757 A EP94920757 A EP 94920757A EP 0707490 A1 EP0707490 A1 EP 0707490A1
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European Patent Office
Prior art keywords
ala
arg
lys
group
homo
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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.)
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EP94920757A
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German (de)
English (en)
Inventor
Ambikaipakan Balasubramaniam
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University of Cincinnati
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University of Cincinnati
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    • 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
    • C07K14/57545—Neuropeptide Y
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P13/00—Drugs for disorders of the urinary system
    • A61P13/02—Drugs for disorders of the urinary system of urine or of the urinary tract, e.g. urine acidifiers
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P15/00—Drugs for genital or sexual disorders; Contraceptives
    • 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
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • A61P9/12—Antihypertensives
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides

Definitions

  • NEUROPEPTIDE Y ANTAGONISTS AND AGONISTS Background of the Invention This invention relates to peptide derivatives which are antagonists or agonists of neuropeptide Y.
  • Neuropeptide Y (NPY)
  • NPY neuropeptide Y
  • Y tyrosine residue
  • tyrosine amide residue at its C-terminus.
  • the peptide was isolated from porcine brain (Tatemoto Proc. Natl . Acad. Sci . U.S.A.
  • NPY elicits several physiological responses by activating specific pre- and post-synaptic receptors. Centrally, NPY is thought to be involved in the regulation of food intake, memory processing and circadian rhythm (Sheikh et al., FEBS Lett . 245: 209-214, 1989). In the periphery, NPY seems to function as a transmitter in sympathetic nerves where it interacts with norepinephrine mainly in the regulation of vasculartone (Sheikh et al. FEBS Lett . 245:209-214, 1989) .
  • NPY receptor subtypes Different structure-activity relationships for NPY analogs in various model systems have indicated that multiple NPY receptor subtypes exist (Michel, Tips 12:389-394, 1991). ahlestedt and coworkers (Reg l . Pept . 13:307-318, 1986) first suggested the existence of two distinct subtypes of NPY receptors. Post-synaptic (Yl-type) effects could be obtained with the complete NPY molecule, while pre-synaptic (Y2-type) effects were found elicited by long C-terminal fragments, as well as with the entire NPY molecule.
  • both Yl and Y2 receptors exhibit nearly equal affinity to NPY and its homologous peptide, peptide YY, but only the Y2 receptors could bind to shorter carboxy1-terminal fragments including NPY(13- 36) as described by Sheikh et al. (FEBS Lett . 245:209- 214, 1989).
  • NPY receptors in rat cardiac ventricular membranes discriminate between NPY and peptide YY but bind NPY(13-36)
  • this system be classified as a subtype of Y2 or a new class (designated Y3) of receptors as discussed below (Balasubramaniam et al. Peptides 11:545-550, 1990).
  • NPY neuropeptide derived from NPY.
  • NPY is also present in high concentrations in a distinct population of nerve fibers innervating the heart and blood vessels ( harton et al., Ann . N. Y. Acad. Sci . 611:133-144, 1990).
  • NPY is now regarded as the predominant peptide present in the cardiovascular system of mammals. This observation has led to numerous studies of the cardiovascular properties of NPY. For example, several investigations have reported that NPY is a potent vasopressor peptide and that it inhibits the coronary blood flow and contractility in isolated perfused hearts (e.g., see Balasubramaniam et al., Regul . Pept . 21:289- 299, 1988; Allen et al. Regul . Pept .
  • NPY is also capable of (1) inhibiting the contractile force of isolated cardiac muscles (Balasubramaniam et al. supra) and myocytes (Piper et al. Nuanyn-Scniedberg ' s Arch . Pharmol . 340: 333-337, 1989) and (2) the adenylate cyclase activity and cAMP production by cardiac muscles (Kassis et al., J. Biol. Chem. 262: 3429-3431, 1987) and myocytes (Kassis et al. supra ; Millar et al.
  • NPY has been implicated in the pathophysiology of a number of diseases including, without limitation, obesity, hypertension and chronic heart failure (CHF) because: (1) hypothalamic NPY levels are elevated in obese rats and decreased in cancer anorectic rats; (2) plasma NPY levels are elevated in CHF and hypertensive patients; (3) negative cardiac inotropic and chronotropic actions; and (4) inhibition of libido and circadian rhythm.
  • CHF chronic heart failure
  • the present invention features compounds having the formula:
  • each each R x and R 2 independently, is H, C 1 -C 12 alkyl
  • a 1 is Tyr, or any aromatic amino acid
  • a 2 is Pro, Hyp, D-Ala, N-Me-Ala, Ac 6 c, D-Pal or Asp
  • a 3 is Ser, Thr, N-Me-Ser, N-Me-Thr, He, Val, Aib, Anb, Nle, or N-Me-Leu;
  • a 4 is the D- or L- isomer selected from the group consisting of Lys, Arg, homo-Arg, diethyl-homo- Arg, Lys- ⁇ -NH-R (where R is H, a branched or straight chain C ⁇ -C ⁇ Q alkyl group, or a C 6 -C 18 aryl group) , or Orn;
  • a 5 is Pro, Hyp, D-Ala, N-Me-Ala, Ac 6 c, D-Pal, or D-Trp;
  • a 6 is Gly or is the D- or L- isomer selected from the group consisting of Asp, Glu, N-Me-Asp, Ala, or Aoc;
  • Y is A ⁇ A S -A ⁇ A ⁇ -A ⁇ - ⁇ -A ⁇ -A ⁇ -A ⁇ -A ⁇ -A ⁇ -A ⁇ -A 18 -
  • a 7 is Asn, Ala, Gin, Gly, or N-Me-Asn;
  • a 8 is Pro, Ser, Thr, Hyp, D-Ala, N-Me-Ala, Ac 5 c, or D-Pal;
  • a 9 is Gly, N-Me-Gly, Ala, or Trp;
  • a 10 is Glu, Asp, N-Me-Glu, Ala, or Nva
  • a 11 is Asp, Glu, N-Me-Asp, Ala, or Anb
  • a 12 is Ala, Nal, Thi , Phe, Bth, Pep, or N-Me-Ala
  • a 13 is Pro, Hyp , D-Ala , N-Me-Ala , Ac 6 c, D-Pal ,
  • a 14 is Ala, Pro, Hyp, D-Ala, N-Me-Ala, Ac 6 c, D-Pal
  • a 15 is Glu, Asp, N-Me-Glu, Ala , or Nva
  • a 16 is Asp, Glu, N-Me-Asp, Ala, or Anb
  • a 17 is Met, Leu, He, Val , Aib, Anb, Nle, or N-Me-Leu;
  • a 18 is Ala, Asn, Gin, Gly, N-Me-Asn, Nal , Thi ,
  • a 19 is the D- or L- isomer selected from the group consisting of Lys, Arg, homo-Arg, diethyl- homo-Arg, Lys-e-NH-R (where R is H, a branched or straight chain ⁇ -C ⁇ alkyl group, or a C 6 -C 18 aryl group) , or Orn;
  • a 20 is Tyr, or any aromatic amino acid;
  • a 21 is Tyr, or any aromatic amino acid;
  • a 22 is Ser, Thr, N-Me-Ser, N-Me-Thr, Ala, Nal,
  • a 23 is Ala, Ser, Thr, Nal, Thi, Phe, Bth, Pep, N-
  • a 24 is Leu, He, Val, Aib, Anb, or N-Me-Leu
  • a 25 is the D- or L- isomer selected from the group consisting of Lys, Arg, homo-Arg, diethyl-homo- Arg, Lys-e-NH-R (where R is H, a branched or straight chain C ⁇ -C ⁇ Q alkyl group, or a C 6 -C 18 aryl group) , or Orn;
  • a 26 is the D- or L- isomer selected from the group consisting of His, Thr, 3-Me-His, ⁇ - pyrazolylalanine, N-Me-His, Lys, Arg, homo-Arg, diethy1-homo-Arg, Lys-e-NH-R (where R is H, a branched or straight chain ⁇ -C ⁇ Q alkyl group, or a C 6 -C 18 aryl
  • each bond can represent either a peptide bond or a pseudopeptide bond, provided that said compound cannot contain more than 3 pseudopeptide bonds, or a pharmaceutically acceptable salt thereof.
  • said pseudopeptide bond is between amino acid residues A 29 -A 30 , A 34 -A 35 , and A 35 -A 36 .
  • Preferred compounds formula (I) include those in which A 32 is D-Trp, D-Phe, D-Tyr, D-Bip, D-Dip, D-Bth, D- Nal, 2-Cl-Trp, Tec, Trp, or a pharmaceutically acceptable salt thereof.
  • the compounds of formula (I) include those in which Y (A 7 -A 24 ) is deleted.
  • the compound of formula (I) is [D-Trp 32 ]NPY, cyclo (2/27) Des-AA 7 " 24 [Asp 2 , D-Ala 6 , D-Lys 27 , D-Trp 32 ]NPY, Des-AA 7 " 24 [D-Ala 5 , Aoc 6 , D-Trp 32 ]NPY, Des-AA 7"24 [D-Ala 5 , Gly 6 , D-Trp 32 ]NPY or Des-AA 7"24 [D-Trp 5 , Aoc 6 , D-Trp 32 ]NPY .
  • the invention features a compound having the formula:
  • X is a chain of 0-7 amino acids, inclusive the N-terminal one of which is bonded to each R 2 and R 2 ; wherein each R x and R 2 , independently, is H, C ⁇ -C. ⁇ alk y! ( e -g « methyl), C 6 -C 18 aryl (e.g., phenyl), C ⁇ C ⁇ acyl (e.g., formyl, acetyl, and myristoyl) , C 7 -C 18 aralkyl (e.g., benzyl), or C 7 -
  • a 27 is the D- or L- isomer selected from the group consisting of any aromatic amino acid, Lys, or a tethered amino acid with an indole ring (e.g., Me-
  • a 28 is Aib or is the D- or L- isomer selected from the group consisting of He, Leu, Val, Anb, Trp, N-Me-Ile, or is deleted;
  • a 29 is Asn, Ala, Gin, Gly, N-Me-Asn, or is deleted;
  • a 30 is Leu, He, Val, Aib, Anb, or N-Me-Leu, or is deleted;
  • a 31 is He, Cys, D-Ala, Leu, Val, Aib, Anb, N-Me-Ile, or is deleted;
  • a 32 is the D- or L- isomer selected from the group consisting of any aromatic amino acid except L- Tyr, a tethered amino acid with an indole ring (e.g., Me-Trp) , Ant, Ser, N-Me-Ser, Thr, N-Me-Thr, Ala, N-Me-Ala, D-Hyp, or any Trp derivative (e.g., 2-chlorotroptophan, or Tec) ;
  • a 33 is the D- or L- isomer selected from the group consisting of Lys, Arg, homo-Arg, diethyl-homo- Arg, Lys-e-NH-R (where R is H, a branched or straight chain C 1 -C 10 alkyl group, or a C 6 -C 18 aryl group) , Orn, or is deleted;
  • a 34 is Gin, Asn, N-Me-Gln, Nle, Nva, Ala, or Gly;
  • a 35 is the D- or L- isomer selected from the group consisting of Lys, Arg, homo-Arg, diethyl-homo- Arg, Lys-e-NH-R (where R is H, a branched or straight chain C 1 -C 10 alkyl group, or a C 6 -C 18 aryl group) , or Orn;
  • a 36 is Tyr, or any aromatic amino acid;
  • W is -OH, -N-R 3 R , or OR 5 (where each R 3 , R 4 , and R 5 , independently, is H, C ⁇ C ⁇ alkyl (e.g., methyl), C 6 -C 18 aryl (e.g., phenyl), C 1 -C 12 acyl (e.g., formyl, acetyl, and myristoyl) , C 7 -C 18
  • Preferred compounds of formula (II) include those where X is A 20 -A 21 -A 22 -A 23 -A 2 -A 25 -A 26 where A 20 is Tyr, or any aromatic amino acid; A 21 is Tyr, or any aromatic amino acid; A 22 is Ser, Thr, N-Me-Ser, or N-Me-Thr; A 23 is Ala, Ser, Thr, Nal, Thi, Phe, Bth, Pep, N- Me-Ala, N-Me-Ser, or N-Me-Thr;
  • a 24 is Leu, He, Val, Aib, Anb, or N-Me-Leu
  • a 25 is the D- or L- isomer selected from the group consisting of Lys, Arg, homo-Arg, diethy1-homo-Arg, Lys-e-NH-R (where R is H, a branched or straight chain C 1 -C 10 alkyl group, or a C 6 -C 18 aryl group) , or Orn;
  • a 26 is the D- or L- isomer selected from the group consisting of His, Thr, 3-Me-His, ⁇ - pyrazolylalanine, N-Me-His, Lys, Arg, homo-
  • R is H, a branched or straight chain C 1 -C 10 alkyl group, or a C 6 -C 18 aryl group
  • R 5 is -OH, -N-R 3 R 4 , or OR 5 (where each R 3 , R 4 , and R 5 , independently, is
  • C ⁇ -C ⁇ acyl e.g., formyl, acetyl, and myristoyl
  • C 7 -C 18 alkaryl e.g., phenyl
  • C ⁇ -C ⁇ acyl e.g., formyl, acetyl, and myristoyl
  • said pseudopeptide bond is between amino acid residues A 29 -A 30 , A 34 -A 35 , and A 35 -A 36 .
  • the compound of formula (II) is [D- Trp 28 , D-Trp 32 ]NPY (27-36), (Des-Asn 29 [D-Trp 28 , D- Trp 32 ]NPY(27-36) , Des-Asn 29 [D-Trp 28 , D-Trp 32 , Nva 34 ]NPY(27- 36), Des-Asn 29 [Trp 28 , Trp 32 , Nva 34 ]NPY(27-36) , and [D- Trp 28 , Ant 32 , Nva 34 ]NPY(27-36) , Des-Asn 29 [D-Trp 28 , Ant 32 , Nva 3 ]NPY(27-36) , or Des-Asn 29 , Arg 33 [D-Trp 28 , Ant 32 , Nva 34 ]NPY(27-36) .
  • the invention features a compound having a formula:
  • each R ⁇ and R 2 independently, is H, C ⁇ C ⁇ alkyl (e.g., methyl), C 6 -C 18 aryl (e.g., phenyl), C- ⁇ C ⁇ acyl (e.g., formyl, acetyl, and myristoyl) ,
  • C 7 -C 18 aralkyl e.g., benzyl
  • a 1 is Tyr, or any aromatic amino acid
  • a 2 is Pro, Hyp, D-Ala, N-Me-Ala, Ac 6 c, D-Pal or Asp
  • a 3 is Ser, Thr, N-Me-Ser, N-Me-Thr, He, Val, Aib, Anb,
  • a 4 is the D- or L- isomer selected from the group consisting of Lys, Arg, homo-Arg, diethy1-homo- Arg, Lys-e-NH-R (where R is H, a branched or straight chain C 1 -C 10 alkyl group, or a C 6 -C 18 aryl group) , or Orn;
  • a 5 is Pro, Hyp, D-Ala, N-Me-Ala, Ac 6 c, D-Pal, or D-Trp;
  • a 6 is Gly or is the D- or L- isomer selected from the group consisting of Asp, Glu, N-Me-Asp, Ala, or
  • Aoc is Cys, Glu, Asn, Ala, Gin, Gly, or N-Me-Asn;
  • a 8 is Pro, Ser, Thr, Hyp, D-Ala, N-Me-Ala, Ac 6 c, or D-
  • a 9 is Gly, N-Me-Gly, Ala, or Trp;
  • Y is A 10 -A 11 -A 12 -A 13 -A 14 -A 15 -A 16 -A 17 or is absent, where
  • a 10 is Glu, Asp, N-Me-Glu, Ala, or Nva
  • a 11 is Asp, Glu, N-Me-Asp , Ala, or Anb;
  • a 12 is Ala , Nal , Thi , Phe , Bth, Pep, or N-Me-Ala;
  • a 13 is Pro, Hyp, D-Ala, N-Me-Ala , Ac 6 c, D-Pal ,
  • a 14 is Ala, Pro, Hyp, D-Ala, N-Me-Ala , Ac 6 c, D-Pal
  • a 15 is Glu, Asp, N-Me-Glu, Ala, or Nva
  • a 16 is Asp, Glu, N-Me-Asp, Ala , or Anb
  • a 17 is Met, Leu, He, Val , Aib, Anb, Nle, or N-Me-Leu
  • a 18 is , Ala, Asn, Gin, Gly, N-Me-Asn, Nal, Thi , Phe, Bth,
  • a 19 is the D- of L- isomer selected from the group consisting of Arg, D-homo-Arg, D-diethy1-homo-Arg, D-Lys-e-NH-R (where R is H, a branched or straight chain C____-C 10 alkyl group, or a C 6 -C 18 aryl group) , or Orn;
  • a 20 is Tyr, or any aromatic amino acid;
  • a 21 is Cys, Lys, Tyr, or any aromatic amino acid;
  • a 22 is Ser, Thr, N-Me-Ser, N-Me-Thr, Ala, Nal, Thi, Phe,
  • a 23 is Ser, Thr, N-Me-Ser, N-Me-Thr, Ala, Nal, Thi, Phe, Bth, Pep, or N-Me-Ala;
  • a 24 is Leu, He, Val, Aib, Anb, or N-Me-Leu;
  • a 25 is the D- or L- isomer selected from the group consisting of Lys, Arg, homo-Arg, diethy1-homo- Arg, Lys-e-NH-R (where R is H, a branched or straight chain C 1 -C 10 alkyl group, or a C 6 -C 18 aryl group) , or Orn;
  • a 26 is the D- or L- isomer selected from the group consisting of His, Thr, 3-Me-His, ⁇ - pyrazolylalanine, N-Me-His, Lys, Arg, homo-Arg, diethy1-homo-Arg, Lys-e-NH-R (where R is H, a branched or straight chain C 1 -C 10 alkyl group, or a C 6 -C 18 aryl group) , or Orn;
  • a 27 is the D- or L- isomer selected from the group consisting of any aromatic amino acid, Lys, or a tethered amino acid with an indole ring (e.g., Me- Trp) ;
  • a 28 is Aib or is the D- or L- isomer selected from the group consisting of He, Leu, Val, Anb, Trp, N-Me-Ile, or is deleted;
  • a 29 is Asn, Ala, Gin, Gly, N-Me-Asn or is deleted;
  • a 30 is Leu, He, Val, Aib, Anb, or N-Me-Leu;
  • a 31 is He, Cys, Leu, Val, Aib, Anb, or N-Me-Ile;
  • a 32 is the D- or L- isomer selected from the group consisting of any aromatic amino acid except L- Tyr, a tethered amino acid with an indole ring
  • Trp 2-chlorotroptophan, or Tec
  • a 33 is the D- or L- isomer is selected from the group consisting of Lys, Arg, homo-Arg, diethyl-homo- Arg, Lys-e-NH-R (where R is H, a branched or straight chain C 1 -C 10 alkyl group, or a C 6 -C 18 aryl group) , Orn, is deleted;
  • a 34 is Gin, Asn, N-Me-Gln, Nle, Nva, Ala, or Gly;
  • a 35 is the D- or L- isomer selected from the group consisting of Lys, Arg, homo-Arg, diethy1-homo- Arg, Lys-e-NH-R (where R is H, a branched or straight chain C j -C- ⁇ g alkyl group, or a C 6 -C 18 aryl group) , or Orn;
  • a 36 is Tyr, or any aromatic amino acid; is -OH, -N-R 3 R 4 , or OR 5 (where R 3 , R 4 , and R 5 , independently, is H, C 1 -C 12 alkyl (e.g., methyl),
  • each bond can represent either a peptide bond or a pseudopeptide bond, provided that said compound cannot contain more than 3 pseudopeptide bonds, or a pharmaceutically acceptable salt thereof.
  • said pseudopeptide bond is between amino acid residues A 29 -A 30 , A 34 -A 35 , and A 35 -A 36 .
  • the compound of formula (III) is cyclo(7/21), Des AA 10"17 [Cys 7 , Cys 21 , D-Trp 32 ]NPY, or cyclo(7/21), Des AA 10-17 [Glu 7 , Lys 21 , D-Trp 32 ]NPY.
  • the invention features a compound with pseudopeptide bonds having the formula:
  • each each R x and R 2 independently, is H, C- j ⁇ C- ⁇ alkyl (e.g., methyl), C 6 -C 18 aryl (e.g., phenyl), C ⁇ C ⁇ acyl (e.g., formyl, acetyl, and myristoyl) , C 7 -C 18 aralkyl (e.g., benzyl), or C 7 -C 18 alkaryl (e.g.
  • a 18 is Ala, Asn, Gin, Gly, N-Me-Asn, Nal, Thi, Phe, Bth, Pep, or N-Me-Ala;
  • a 19 is the D- or L- isomer selected from the group consisting of Lys, Arg, homo-Arg, diethyl-homo- Arg, Lys-e-NH-R (where R is H, a branched or straight chain C. j _-C 10 alkyl group, or a C 6 -C 18 aryl group) , or Orn;
  • a 20 is Tyr, or any aromatic amino acid;
  • a 21 is Tyr, or any aromatic amino acid;
  • a 22 is Ser, Thr, N-Me-Ser, N-Me-Thr, Ala, Nal, Thi, Phe, Bth, Pep, or N-Me-Ala
  • a 23 is Ser, Thr, N-Me-Ser, N-Me-Thr, Ala, Nal, Thi, Phe,
  • a 24 is Leu, He, Val, Aib, Anb, or N-Me-Leu
  • a 25 is the D- or L- isomer selected from the group consisting of Lys, Arg, homo-Arg, diethy1-homo- Arg, Lys-e-NH-R (where R is H, a branched or straight chain C 1 -C 10 alkyl group, or a C 6 -C 18 aryl group) , or Orn;
  • a 26 is the D- or L- isomer selected from the group consisting of His, Thr, 3-Me-His, ⁇ - pyrazolylalanine, N-Me-His, Lys, Arg, homo-Arg, diethy1-homo-Arg, Lys-e-NH-R (where R is H, a branched or straight chain C ⁇ C ⁇ alkyl group, or a C 6 -C 18 aryl group) , or Orn;
  • a 33 is the D- or L- isomer selected from the group consisting of Lys, Arg, homo-Arg, diethy1-homo- Arg, Lys-e-NH-R (where R is H, a branched or straight chain C_ j _-C 10 alkyl group, or a C 6 -C 18 aryl group) , Orn, or is deleted;
  • a 34 is Gin, Asn, N-Me-Gln, Nle, Nva, Ala, or Gly;
  • a 35 is the D- or L- isomer selected from the group consisting of Lys, Arg, homo-Arg, diethyl-homo-
  • Arg, Lys-e-NH-R (where R is H, a branched or straight chain C 1 -C 10 alkyl group, or a C 6 -C 18 aryl group) , or Orn;
  • a 36 is Tyr, or any aromatic acid; is -OH, -N-R 3 R 4 , or OR 5 (where each R 3 , R 4 , and R 5 , independently, is H, C ⁇ -C ⁇ alkyl (e.g., methyl), C 6 -C 18 aryl (e.g., phenyl), C j -C- ⁇ acyl (e.g., formyl, acetyl, and myristoyl) , C 7 -C 18 aralkyl (e.g., benzyl), or C 7 -
  • each bond can represent either a peptide or a pseudopeptide bond, provided that said compound cannot contain more than 3 pseudopeptide bonds, or a pharmaceutically acceptable salt thereof.
  • the compound contains a pseudopeptide bond between A 30 and A 31 ; A 31 and A 32 ; or A 32 and A 33 .
  • the invention features a method of suppressing an NPY mediated physiological response in a tissue other than the heart in a subject comprising administering to said subject a compound having the following formula:
  • each each R x and R 2 independently, is H, C ⁇ C ⁇ alkyl (e.g., methyl), C 6 -C 18 aryl (e.g., phenyl), C ⁇ C ⁇ acyl (e.g., formyl, acetyl, and myristoyl), c 7 " i 8 aralkyl (e.g., benzyl), or C 7 -C 18 alkaryl (e.g., p-methylpheny1) ;
  • a 18 is Ala, Asn.
  • a 19 is the D- or L- isomer selected from the group consisting of Lys, Arg, homo-Arg, diethyl-homo- Arg, Lys-e-NH-R (where R is H, a branched or straight chain C ⁇ C- j ⁇ alkyl group, or a C 6 -C 18 aryl group) , or Orn;
  • a 20 is Tyr, or any aromatic amino acid;
  • a 21 is Tyr, or any aromatic amino acid;
  • a 22 is Ser, Thr, N-Me-Ser, or N-Me-Thr;
  • a 23 is Ala, Ser, Nal, Thi, Phe, Bth, Pep, or N-Me-Ala;
  • a 24 is Leu, He, Val, Aib, Anb, or N-Me-Leu;
  • a 25 is the D- or L- isomer selected from the group consisting of Lys, Arg, D-homo-Arg, D-diethyl- homo-Arg, D-Lys-e-NH-R (where R is H, a branched or straight chain C- j -C jg alkyl group, or a C 6 -C 18 aryl group) , or Orn;
  • a 26 is the D- or L- isomer selected from the group consisting of His, Thr, 3-Me-His, ⁇ - pyrazolylalanine, N-Me-His, Lys, Arg, homo-Arg, diethy1-homo-Arg, Lys-e-NH-R (where R is H, a branched or straight chain C 1 -C 10 alkyl group, or a C 6 -C 18 aryl group) , or Orn;
  • a 27 is the D- or L- isomer selected from the group consisting of any aromatic amino acid,
  • a 28 is Aib or is the D- or L- isomer selected from the group consisting of He, Leu, Val, Anb, Trp, N-Me-Ile, or is deleted;
  • a 29 is Asn, Ala, Gin, Gly, N-Me-Asn, or is deleted;
  • a 30 is Leu, He, Val, Aib, Anb, or N-Me-Leu;
  • a 31 is He, Cys, Leu, Val, Aib, Anb, or N-Me-Ile;
  • a 32 is the D- or L- isomer selected from the group consisting of any aromatic amino acid except L- Tyr, a tethered amino acid with an indole ring (e.g., Me-Trp) , Ant, Ser, N-Me-Ser, Thr, N-Me-Thr, Ala, N-Me-Ala, D-Hyp, or any Trp derivative (e.g., 2-chlorotroptophan, or Tec) ;
  • a 33 is the D- or L- isomer selected from the group consisting of Lys, Arg, homo-Arg, diethy1-homo- Arg, Ly
  • a 36 is Tyr, or any aromatic acid
  • W is -OH, -N-R 3 R 4 , or OR 5 (where each R 3 , R 4 , and R 5 , independently, is
  • each bond can represent either a peptide bond or a pseudopeptide bond, provided that said compound cannot contain more than 3 pseudopeptide bonds, or a pharmaceutically acceptable salt thereof.
  • said pseudopeptide bond is between amino acid residues A 29 -A 30 , A 34 -A 35 , and A 35 -A 36 . or a pharmaceutically acceptable salt thereof.
  • the method suppresses the activity of the NPY (Y-l) receptor or the NPY (Y-2) receptor.
  • the invention features a method of suppressing a NPY(Y-l) receptor mediated physiological response in the hypothalamus of a subject comprising administering to said subject the compound of formula
  • the invention features a method of suppressing the blood pressure of a subject experiencing hypertension which comprises administering to said subject the compound of formula (I) .
  • the invention features a method of suppressing a NPY(Y-3) receptor mediated physiological response in the cardiovascular system of a subject comprising administering to said subject the compound of formula (IV) .
  • a therapeutically effective amount of a compound of formula (I) , (II) , (III) or (IV) and a pharmaceutically acceptable carrier substance, e.g., magnesium carbonate or lactose, together form a therapeutic composition capable of suppressing an NPY mediated physiological response.
  • This composition can be in the form a pill, tablet, capsule, liquid, or sustained released tablet for oral administration; or a liquid for nasal administration as drops or spray; or a liquid for intravenous, subcutaneous, parenteral, or intraperitoneal administration.
  • compositions for intramuscular administration to a subject in need of the composition.
  • the composition includes a lipophilic salt and is suitable for administration in the form of an oil emulsion or dispersion to a subject in need of the composition.
  • the invention features methods for suppressing an NPY mediated physiological response in a subject; such methods involve administering one or more of the above mentioned compounds to a subject in a dosage effective to lower blood pressure; to suppress the appetite; to augment the libido; to stimulate cardiovascular function; on to modulate the circadian rhythm.
  • the invention features methods for stimulating an NPY mediated physiological response in a subject; such methods involve administering one or more of the above mentioned compounds to a subject in a dosage effective to increase blood pressure; to increase the appetite; to augment the libido; or to stimulate cardiovascular function.
  • R is -CH 2 COOH for Asp
  • R is -H for Gly
  • R is - CH 2 OH for Ser
  • R is -CH 3 for Ala
  • R is -CH 2 CH 2 CH 2 CH 2 NH 2 for Arg.
  • pseudopeptide bond is meant that the carbon atom participating in the bond between two residues is reduced from a carbonyl carbon to a methylene carbon, i.e., CH 2 - NH; or less preferably that of C)-NH is replaced with any Of CH 2 -S, CH 2 -0, CH 2 -CH 2 , CH 2 -CO, or CH 2 -CH 2 .
  • a pseudopeptide peptide bond is symbolized herein by or " ⁇ ".
  • the compounds of Formulae (I), (II), (HI), or (IV) are cyclic.
  • the cyclization is formed by a disulfide or lactam bridge (amide bond) .
  • the disulfide or amide bond which links two residues in a compound of the invention are formed between the side chain functionalities. That is, between the side-chain carboxyl group of an acidic amino acid residue (e.g.. Asp or Glu) and the side chain amino group of a basic amino acid residue (e.g., Lys or Orn), or between the side chain sulfhydryl groups of two Cys.
  • the amide or disulfide bond between two residues are not shown.
  • a compound of this invention is also denoted by another format, e.g. cyclo (2/27) Des- AA 7 - 24 [Asp 2 , D-Ala 6 , D-Lys 27 , D-Trp 32 ] NPY and cyclo(7/21) Des AA 10"17 [Cys 7 , Cys 21 , D-Trp 32 ]NPY.
  • Preferred cyclic compounds of the invention are cyclo (2/27) Des AA 7 ⁇ 24 [Asp 2 , D-Ala 6 , D-Lys 27 , D-Trp 32 ] NPY and cyclo(7/21) Des AA 10_17 [Cys 7 , Cys 21 , D-Trp 32 ]NPY.
  • the invention features novel dimeric analogs of NPY.
  • the dimer may be formed by either including one compound of Formula I, II, II, or IV and one compound of Formula I, II, III, or IV.
  • the dimer is formed by utilizing a dicarboxylic acid linker capable of binding to a free amine, either primary or secondary, located within each compound. See R.
  • the dimer is formed by utilizing an amino acid linker capable of binding to a free amine group of one compound and a free carboxylic acid group of the other compound.
  • the amino acid linker is a non- ⁇ -amino acid.
  • suitable amino acid linkers are amino-caproic acid and amino-valeric acid.
  • the dimer is formed by disulfide bridge between cysteines located within each compound. See M. Berngtowicz and G. Piatsueda, Peptides: Structure and Function 233-244 (Pierce Chemical Co. 1985); F. Albericio, et al., Peptides 1990 535 (ESCOM 1991) .
  • Preferred dimeric compounds of the invention are Bis(31/31) [Cys 31 , Trp 32 , Nva 34 ]NPY(27-36) , and Bis(31/31) (Cys 31 , Trp 32 , Nva 34 ]NPY(31-36) ,
  • Tic tetrahydroisoquinoline-3-carboxylic acid
  • Aib aminoisobutyric acid
  • the compounds of the invention are useful for reducing, suppressing or mitigating the effects of NPY.
  • the compounds of the invention are especially useful in treating any number of illnesses that involve eating disorders, cardiovascular function, alterations in sexual function, as well as disorders of sleep and circadian rhythms (see, e.g., Harrison 's
  • NPY antagonists as described herein, allows for the selective antagonism of different classes of NPY receptors, e.g., Y3 cardiac receptors, without adverse interaction with other NPY receptors.
  • the compounds are also useful for stimulating NPY receptor mediated events, e.g., increasing the blood pressure of a subject.
  • DRAWINGS Fig. 1 shows the comparison of the effects of D- Trp or D-Trp(CHO) substituted NPY analogs (1.0 ⁇ M) on the isoproternol stimulated adenylate cyclase activity of rat hypothalmic membranes. Iso, isoproternol.
  • Fig. 2 shows the displacement of 125 I-NPY bound to rat hypothalamic membranes by increasing concentrations NPY (•) and [D-Trp 32 ] NPY (D) .
  • Fig. 3 shows the dose-response effects of increasing concentrations of [D-Trp 32 ] NPY (D) , NPY alone (•) ; NPY in the presence of 30 (A) and 300 (I) nM doses of [D-Trp 32 ] NPY on the isoproterenol stimulated adenylate cyclase activity of rat hypothalamic membranes.
  • Fig. 4 shows the comparison of the effects of [D- Trp 32 ]NPY (1.0 ⁇ M) on the inhibition of isoproterenol stimulated adenylate cyclase activity of rat hypothalamic membranes by NPY (100 nM) and serotonin (100 nM) .
  • a p ⁇ 0.01 compared to isoproterenol;
  • b not significant compared to isoproterenol .
  • Fig. 5 shows the antagonism of NPY induced feeding in rats by [D-Trp 32 ]NPY.
  • Fig. 7 shows the effects of increasing concentrations of NPY in the absence (0) and presence (•) of Des-AA 7"2 [D-Ala 5 , Aoc 6 , D-Trp 32 ] NPY (1 ⁇ M) on the isoproterenol stimulated cAMP production by SK-N-MC cells. Also shown is the effect of increasing concentrations of Des-AA 7"2 [D-Ala 5 , Aoc 6 , D-Trp 32 ]NPY (D) on the isoproterenol stimulated cAMP production by SK-N- MC cells.
  • Fig. 7 shows the effects of increasing concentrations of NPY in the absence (0) and presence (•) of Des-AA 7"2 [D-Ala 5 , Aoc 6 , D-Trp 32 ] NPY (1 ⁇ M) on the isoproterenol stimulated cAMP production by SK-N-MC cells. Also shown is the effect of increasing concentrations of Des-
  • NPY 8 shows the effects of increasing concentrations of NPY on the blood pressure of anesthetized rats in the absence (0) and presence (•) of 200 nmol/kg of Des-AA 7"24 [D-Ala 5 , Aoc 6 , D-Trp 32 ]NPY.
  • Fig. 9 shows the effects of increasing concentrations of NPY (o) and NPY (18-36) ( ⁇ ) on the binding of 125 I-NPY to SK-N-BE2 cells.
  • Fig. 10 shows the effects of NPY (O) , NPY (18-36) ( ⁇ ) and NPY in the presence of l ⁇ M dose of NPY (18-36) (•) on forskolin stimulated cAMP production by SK-N-BE2 cells.
  • Figs. 11A-11C show the analytical RPLC of [ ⁇ 30 - 3l ] NPY (18-36) (HA), [ ⁇ 32 - 33 j Np ⁇ ( ⁇ -36) (11B) , and [ ⁇ 33 ⁇ 34 ] NPY (18-36) (HC).
  • Fig. 12 shows the inhibition of 125 I-NPY binding to rat cardiac ventricular membrane by NPY (0) , NPY (18-36) ( ⁇ ) , [ ⁇ 30 / 31 ] NPY (18-36) ( ⁇ ) , [ ⁇ 1 / 32 -j NP y (18-36) ( A ), and [ ⁇ 32 / 33 ] N py (is-36) (D) .
  • the analogs of the invention have the general formula recited in the Summary of the Invention aJove.
  • the analogs of the invention are based upon the biologically active full-length molecule (amino acids 1- 36) comprising amino acids of NPY and PYY and derivatives thereof; and upon the biologically active subfragments comprising amino acids of NPY and PYY and derivatives thereof.
  • the analogs of the invention may have one or more modifications to the NPY and PYY sequences (see aiove) .
  • the compounds may have one or more of the following modifications which are useful for obtaining selective activity at a NPY receptor: a D-Trp or Aoc or D-Ala in place of one or two or three natural amino acids; or a deletion of several N-terminal amino acids; or the introduction of a pseudopeptide bond instead of a peptide bond between two adjacent amino acids.
  • the analog is capable of acting as a competitive inhibitor of the naturally occurring NPY peptide by binding to the receptor and, by virtue of one of the modifications described supra herein, fail to exhibit the biological activity of the naturally occurring peptide.
  • the peptides for which introduction of a pseudopeptide bond between two residues, or the replacement of one or more natural amino acids with a D- Trp, or the deletion ("des") of the N-terminal residues or internal residues are useful in activity associated NPY activity.
  • the analogs of the invention can be provided in the form of pharmaceutically acceptable salts.
  • preferred salts are those with therapeutically acceptable organic acids, e.g., acetic, lactic, maleic, citric, malic, ascorbic, succinic, benzoic, or pamoic acid, as wells as polymeric acids and slats with inorganic acids such as the hydrohalic acids, e.g., hydrochloric and sulfuric acids.
  • the compounds of the present invention i.e., compounds of formulas (I) , (II) , (III) , (IV) , and (V) may be synthesized by any techniques that are known to those skilled in the peptide art. Such techniques are described in, e.g.. Solid Phase Peptide Synthesis , eds, John M. Stewart and Janis D. Young, Pierce Chemical Company, Rockford, IL, 2nd edition.
  • pseudopeptide bonds may, if desired, may be introduced at various positions, e.g., between amino acid residues 31-32 of NPY(18-36) or between residues 32-33 of NPY(18-36), or of any peptide as described below.
  • optically pure Boc-AA-CHO can be obtained in good yields and coupled directly to the ⁇ -NH 2 group of the peptide resin by published methods (Sasaki et al., Peptides 8:119-121, 1987; Fehrentz et al..
  • any suitable in vivo or in vitro system may be utilized to assay and test the effectiveness of the compounds of the invention.
  • Such assays may employ in vivo methods for evaluating physiological responses, e.g., blood pressure, renovascular function, feeding behavior, or circadian rhythm, or in vivo biochemical systems evaluating receptor binding in a suitable cell line, e.g., SK-N-MC (ATCC#HBT 10) or SK-N-BE(2) (Barnes et al. In Vitro 17: 619-631, 1981); or in isolated cells, e.g., cells isolated from the spleen, kidney, heart or brain.
  • SK-N-MC ATCC#HBT 10
  • SK-N-BE(2) Barnes et al. In Vitro 17: 619-631, 1981
  • isolated cells e.g., cells isolated from the spleen, kidney, heart or brain.
  • NPY receptors e.g. the Y-l, Y-2, and Y-3 receptor categories. Described below are assay methods which can be utilized with cell lines such as SK-N-MC and SK-N-BE2 or isolated cardiac membranes which possess the high-affinity NPY receptor sites Y-l, Y-2, and Y-3, respectively.
  • cell lines such as SK-N-MC and SK-N-BE2 or isolated cardiac membranes which possess the high-affinity NPY receptor sites Y-l, Y-2, and Y-3, respectively.
  • Other systems are also known for evaluating NPY antagonists to the Y-l receptor, e.g. VSM cells (Sheikh et al., Am . J. Physiol .
  • HEL cells Motulsky et al. Amer. J. Physiol . 255: E880-E885, 1988
  • Y-2 receptor e.g., kidney (Sheikh et al.. Am. J. Physiol 26:F978-F984) , spleen (Lunberg et al., Eur. J. Pharmal. 145:21-29, 1988), dorsal root ganglion (Bleakman et al., Br. J. Pharmal. 103:1781-1789, 1991) and hippocampal cells (Sheikh et al., J. Biol. Chem.
  • Y-3 receptors e.g., in cardiac ventricular membranes (Balasubramaniam et al.. Peptides 11: 545-550, 1990) , chro affin cells, rat gastric mucosa (Michel, M.C., Trends in Pharmol . Sci . 12: 389-394, 1991) and brain stem.
  • the ability of the compounds of the invention to act as antagonists of NPY can be demonstrated by any number of methods known in the art.
  • the compounds can be shown to compete with iodinated neuropeptide Y for receptors using the methods described by Lundberg et al. (Eur. J. Pharmol . 145: 21-29, 1988); Gordon et al. (J. Neurochemistry 55:506-513, 1990); Walker et al. (Mol . Pharmacol . 34:779-792, 1988); Balasubramaniam et al . (Peptides 10:1283-1286, 1989), and others.
  • rat hypothalamus was isolated and the membranes were prepared for binding and adenylate cyclase studies according to standard methods (Unden et al. 1984. Eur. J. Biochem 145: 525-530; Westlind-Danielsson et al. 1987. Neurosci . Lett . 74: 237-242). Displacement studies were performed in a total volume of 0.25 ml 20 mM HEPES buffer, pH 7.4, containing 1% bovine serum albumin, 0.1% bacitracin, 300 ⁇ m PMSF and 5 KlU/ml aprotinin.
  • Adenylate cyclase activity of the hypothalamic and cerebral cortex membranes was determined by incubating 50 ⁇ g of membranes in a total volume of 0.20 ml Tris-HCL 30 mM pH 7.4 buffer containing 150 mM NaCl, 8.25 mM MgCl 2 , 0.75 mM EGTA, 1.5 theophylline, 20 ⁇ g/ml aprotinin, 100 ⁇ g/ml bovine serum albumin, 1 mM ATP, 20 mM creatine phosphate, 1 mg/ml phosphocreatine kinase, 10 ⁇ M isopreternol, 10 ⁇ M GTP, and various concentrations of peptides (0-10 ⁇ M) .
  • rat cardiac ventricular membranes and iodination of NPY were prepared according to the method described by Balasubramaniam et al. (Peptides 11: 545- 550, 1990) . Displacement studies were performed in a total volume of 0.25 ml of 20 mM HEPES assay buffer, pH 7.6, containing 2% bovine serum albumin, 100 ⁇ M phenylmethylsulfonyl fluoride, 4 ⁇ g/ml leupeptin, 4 ⁇ g/ml chymostatin, 5 kallikrein-inactivating units/ml aprotinin, and 0.1% bacitracin.
  • Adenylate cyclase activity was measured by Rosselin et al. (Biochim . Biophys . Acta 304:541-551, 1977) . Each experiment was carried out in a total volume of 200 ⁇ l solution containing 30 mM Tris-HCl, pH 7.4, 150 mM NaCl, 8.25 mM MgCl 2 0.75 mM EGTA, 1.5 mM theophylline, 20 ⁇ g/ml aprotinin, 100 ⁇ g/ml bacitracin, 1 mg/ml BSA, 1 mM ATP, 20 mM creatine phosphate, 1 mg/ml phosphocreatine kinase, 10 ⁇ M isoproterenol, 10 ⁇ M GTP, and various concentrations of peptides (0-10 ⁇ M) .
  • Any suitable m vivo model system can be used to evaluate the antagonistic properties of the compounds of the invention.
  • Such models include those used to evaluate feeding and memory behavior (Flood et al., Peptides 10:963-966), and vasoconstriction and hypertension (Balasubramaniam et al. Biochim et Biophys Acta 997: 176-188, 1989).
  • feeding studies were performed using Spraque Dawley rats (350-450 g) with paraventricular hypothalamic cannulae to investigate effects of NPY analogs (Chance et al. 1989. Peptides 10: 1283-1286) .
  • Antagonism of NPY induced feeding in rats was by [D-Trp 32 ]NPY.
  • Groups of rats received intrahypothalamic injections (l ⁇ l) of artificial CSF or 10 ⁇ g of [D-Trp 32 ]NPY.
  • [D-Trp 32 ]NPY and its formulated derivative did not exhibit significant inhibitory effect on adenylate cyclase activity at this concentration.
  • NPY and [D-Trp 32 ]NPY inhibited 125 I-NPY bound to rat hypothalamic membranes in a dose-dependent manner with IC 50 values of 0.63 nM and 3.0 nM, respectively. It is this high receptor activity and the complete loss of intrinsic activity that suggests that [D-Trp 32 ]NPY may be an antagonist of NPY in rat hypothalamus.
  • NPY hypothalamic NPY has been shown to elicit a feeding response
  • [D- Trp 32 ]NPY has been shown to elicit a feeding response
  • Fig. 5 shows that intrahypothalamic injection of NPY (1 ⁇ g) significantly (p ⁇ 0.01) stimulated the cumulative food intake as compared to vehicle (artificial cerebrospinal fluid) treatment over 1 hr.
  • [D-Trp 32 ]NPY (1 ⁇ g) did not stimulate feeding significantly over this period, nor did it attenuate NPY (1 ⁇ g) - induced feeding at this concentration.
  • Des-AA 7"24 [D-Ala 5 , Aoc 6 , D-Trp 32 ]NPY surprisingly exhibited moderate affinity (Table I) , and its presence (1.0 ⁇ M) shifted the inhibitory dose- response curve of NPY on SK-N-MC cAMP production parallel to the right (Fig. 7) .
  • NPY-induced anorectic rats we tested the effects on NPY-induced anorectic rats.
  • Fig. 8 shows that NPY doses of 0.1, 1.0 and 10.0 nmol/kg, during baseline, increased systolic blood pressure (SBP) by 8+7, .26+6 and 37+7 mmHg respectively.
  • SBP systolic blood pressure
  • NPY doses of 0.1, 1.0 and 10.0 nmol/kg increased SBP by 4+5, 9+5 and 29+17 mmHg respectively.
  • Des-Asn 29 [D-Trp 28 ' 32 ]NPY(27-36) bound with moderate potency to Y-l receptors, and also did not exhibit any intrinsic activity on isoproterenol stimulated cAMP production by SK-N-MC cells. These observations suggest that Des-Asn 29 [D-Trp 28 ' 32 ]NPY(27-36) or its analogs will prove useful for the development low molecular weight selective antagonist compounds for Y-l receptors. TABLE I Peptides IC 50 1 ⁇ ) for tne inhibition of
  • NPY receptor Y-2 activity may also be assayed and tested for NPY receptor Y-2 activity using the methods described supra .
  • a compound e.g., [D- Trp 32 ]NPY
  • NPY(18-36) previously shown to be an antagonist of NPY in rat cardiac membranes bearing Y-3 receptors, antagonizes the inhibitory effect on the cAMP production of SK-N-BE2 cells bearing Y-2 receptor subtypes as shown in Figures 9 and 10.
  • NPY RECEPTOR (Y-3 SUBTYPE)
  • Table II shows the results for the increased affinity and selectivity of pseudopeptide analogs of NPY(18-36) for Y-3 receptors.
  • the introduction of pseudobonds (-CH 2 NH-) at positions 31-32 or 32-33 of NPY(18-36) was found to substantially increase Y-3 receptor affinity (see Table 2) .
  • PEPTIDES IC 50 (nM) for the inhibition of 125 I-NPY binding to:
  • reaction vessel 1.0 g peptide resin, 0.8 g p-cresol, 0.2g thiocresol, 0.8 ml (CH 3 ) 2 and 5 ml HF were stirred for 40 min of reaction and an additional 60 min. of HF evacuation. During these procedures temperature of reaction vessel was kept between 0°C - 4°C. Then the peptide resin was transferred into a fitted filter funnel in Et 2 0 and washed with excess of Et 2 0. Free peptide was extracted with 30% HOAc (2x15ml) . Peptide solution was diluted to 10% HOAc (60ml H 2 0) and lyophilized. 390 mg crude peptide was obtained from this procedure.
  • Peptide synthesis was as described above using an Automated ABI 430A synthesizer.
  • the free peptide was obtained by treating the protected peptide resin (l.Og) with HF (10 ml) containing dimethyl sulfide (0.8 ml), p- cresol (0.2g) for 1 h at -2 to -4 C.
  • the residue was transferred to a fitted filter funnel with diethyl ether, washed repeatedly with diethyl ether, and the peptide extracted with 10% HOAC(2X 15 ml) and lyophilized.
  • the crude peptide (lOOmg) thus obtained was dissolved in 6M guanidine HCL (6 ml) diluted with 500 ml of distilled water and the pH adjusted to 8 with ammonia.
  • a solution of potassium ferricyanide (1% w/v) was gradually added with constant stirring until a yellow color persisted.
  • the pH of the solution was adjusted to 5 with acetic acid and the solution stirred with an anion exchange resin (AG-3, Cl- form, lOg wet weight) for 30 min, passed through a 0.45 microns filter, and pumped into a semipreparative column (250X10 mm), washed with 0.1%TFA-H 2 0 until a flat base line was obtained.
  • the column containing the peptide was then subjected to gradient elution as described for NPY, and the purified peptide was characterized by amino acid and mass spectral analysis.
  • NPY is a potent vasoconstrictor and or orexigenic agent, as well as an inhibitor of libido and effector of circulation rhythm, it is likely that the administration of one or more compounds of the invention may suppress or inhibit the deleterious effects of NPY. Therefore, the NPY antagonists of the invention are suitable for the treatment of any number of diseases related to cardiovascular function (e.g., congestive heart failure or hypertension) , obesity, anorexia, blood pressure, asthma, pulmonary hypertension, renal hypertension, memory retention, sexual dysfunction (e.g. impotence) , and disorders involving sleep and circadian rhythms.
  • cardiovascular function e.g., congestive heart failure or hypertension
  • anorexia blood pressure
  • asthma pulmonary hypertension
  • renal hypertension e.g. pulmonary hypertension
  • memory retention e.g. impotence
  • sexual dysfunction e.g. impotence
  • the compounds of formula (I) , (II) , (III) are useful for treating for controlling feeding disorders and blood pressure; the compounds of formula (IV) are useful for treating any number of heart ailments, e.g., chronic heart failure, as well as promoting recovery from ischemia since the compounds are expected to enhance myocardium contraction; and the compounds of formula (IV) are useful for controlling NPY actions mediated by Y-2 receptor subtypes, e.g., for controlling the effects of NPY on renal blood flow, glomerular filtration rate, natriuresis and renin secretion.
  • the appropriate NPY antagonist is administered as a therapeutic preparation (as described below) in accordance with the condition to be treated.
  • an effective amount of an NPY antagonist e.g., ⁇ 30 ⁇ 31 NPY(18-36)
  • an NPY antagonist e.g., ⁇ 30 ⁇ 31 NPY(18-36)
  • an effective amount of an NPY antagonist is administered via any of the usual and acceptable methods known in the art, either singly or in combination with another compound or compounds of the present invention.
  • These compounds or compositions can thus be administered orally, sublingually, parenterally (e.g., intramuscularly, intravenously, subcutaneously, or intradermally) or by inhalation, and in the form or either solid, liquid or gaseous dosage, including tablets and suspensions.
  • the administration can be conducted in a single unit dosage form with continuous therapy or in a single dose therapy ad libitum.
  • the dose of the compound of the present invention for treating the above-mentioned disorders varies depending upon the manner of administration, the age and the body weight of the subject, and the condition of the subject to be treated, and ultimately will be decided by the attending physician or veterinarian.
  • Such amount of the active compound as determined by the attending physician or veterinarian is referred to herein as a "therapeutically effective amount".
  • a typical administration is oral administration or parenteral administration.
  • the daily dose in the case of oral administration is typically in the range of 0.1 to 100 mg/kg body weight, and the daily dose in the case of parenteral administration is typically in the range of 0.001 to 50 mg/kg body weight.
  • the therapeutic agents be relatively non-toxic, non-antigenic and non-irritating at the levels in actual use.

Abstract

L'invention décrit des analogues se comportant comme des antagonistes et des agonistes du NPY, ainsi que des méthodes d'utilisation de ces composés, permettant de contrôler une activité biologique telle que l'appétit et la fonction cardio-vasculaire.
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Families Citing this family (36)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5516653A (en) * 1993-12-28 1996-05-14 Synaptic Pharmaceutical Corporation DNA encoding a human neuropeptide Y/peptide YY/pancreatic polypeptide receptor (Y4) and uses thereof
US5545549A (en) 1994-02-03 1996-08-13 Synaptic Pharmaceutical Corporation DNA encoding a human neuropeptide Y/peptide YY (Y2) receptor and uses thereof
US5989920A (en) 1994-12-02 1999-11-23 Synaptic Pharmaceutical Corporation Methods of modifying feeding behavior compounds useful in such methods and DNA encoding a hypothalmic atypical neuropeptide Y/peptide YY receptor Y5
US5602024A (en) 1994-12-02 1997-02-11 Synaptic Pharmaceutical Corporation DNA encoding a hypothalamic atypical neuropeptide Y/peptide YY receptor (Y5) and uses thereof
EP0759441A3 (fr) * 1995-06-30 1999-06-30 Eli Lilly And Company Procédés de traitement des pathologies associées aux peptides neuropeptide Y
ZA965346B (en) * 1995-06-30 1997-12-24 Lilly Co Eli Methods of treating neuropeptide Y-associated conditions.
US5943242A (en) 1995-11-17 1999-08-24 Pact Gmbh Dynamically reconfigurable data processing system
FR2754709B1 (fr) 1996-10-23 1999-03-05 Sanofi Sa Composition cosmetique contenant un antagoniste des recepteurs du neuropeptide gamma et alpha 2 antagonistes susceptibles d'etre incorpores dans une telle composition
CA2274594C (fr) * 1996-12-16 2006-10-10 Banyu Pharmaceutical Co., Ltd. Derives d'aminopyrazole
US6542998B1 (en) 1997-02-08 2003-04-01 Pact Gmbh Method of self-synchronization of configurable elements of a programmable module
CN100378118C (zh) 1997-04-15 2008-04-02 Csir公司 具有食欲抑制剂活性的药物组合物
US6713265B1 (en) 1997-06-04 2004-03-30 Synaptic Pharmaceutical Corporation Methods of modifying feeding behavior, compounds useful in such methods, and DNA encoding a hypothalamic atypical neuropeptide Y/peptide YY receptor (Y5)
GB2355657B (en) 1999-10-27 2004-07-28 Phytopharm Plc Inhibitors Of Gastric Acid Secretion
GB2363985B (en) 2000-06-30 2004-09-29 Phytopharm Plc Extracts,compounds & pharmaceutical compositions having anti-diabetic activity and their use
GB0217015D0 (en) * 2002-07-23 2002-08-28 Bioacta Ltd Peptide 4
WO2004098591A2 (fr) 2003-05-05 2004-11-18 Probiodrug Ag Inhibiteurs de glutaminyl-cyclase
EP1680120A2 (fr) 2003-11-03 2006-07-19 Probiodrug AG Combinaisons utiles au traitement de troubles neuronaux
CN1918131B (zh) 2004-02-05 2011-05-04 前体生物药物股份公司 谷氨酰胺酰基环化酶抑制剂
WO2007097742A1 (fr) * 2006-02-21 2007-08-30 University Of Cincinnati Méthodes en rapport avec des récepteurs induits par des corticoïdes
WO2008055945A1 (fr) 2006-11-09 2008-05-15 Probiodrug Ag Dérivés 3-hydr0xy-1,5-dihydr0-pyrr0l-2-one utiles en tant qu' inhibiteurs de la glutaminyl-cyclase dans le traitement des ulcères, du cancer et d'autres maladies
SI2091948T1 (sl) 2006-11-30 2012-07-31 Probiodrug Ag Novi inhibitorji glutaminil ciklaze
EP2481408A3 (fr) 2007-03-01 2013-01-09 Probiodrug AG Nouvelle utilisation d'inhibiteurs glutaminyle cyclase
DK2142514T3 (da) 2007-04-18 2015-03-23 Probiodrug Ag Thioureaderivater som glutaminylcyclase-inhibitorer
CA2751636C (fr) * 2009-02-20 2016-08-30 Ipsen Pharma S.A.S. Analogues de neuropeptide y ayant au moins une substitution par un acide amine synthetique
AU2010216385B2 (en) * 2009-02-20 2013-06-20 Ipsen Pharma S.A.S. Analogues of neuropeptide Y having proline substitution at position 34
CN102325545A (zh) 2009-02-20 2012-01-18 益普生制药股份有限公司 具有神经肽y受体结合化合物的细胞毒性缀合物
AU2010294214B2 (en) 2009-09-11 2015-05-07 Vivoryon Therapeutics N.V. Heterocylcic derivatives as inhibitors of glutaminyl cyclase
JP6026284B2 (ja) 2010-03-03 2016-11-16 プロビオドルグ エージー グルタミニルシクラーゼの阻害剤
AU2011226074B2 (en) 2010-03-10 2015-01-22 Vivoryon Therapeutics N.V. Heterocyclic inhibitors of glutaminyl cyclase (QC, EC 2.3.2.5)
EP2560953B1 (fr) 2010-04-21 2016-01-06 Probiodrug AG Inhibiteurs de glutaminyl cyclase
JP6050264B2 (ja) 2011-03-16 2016-12-21 プロビオドルグ エージー グルタミニルシクラーゼの阻害剤としてのベンゾイミダゾール誘導体
EP3337788A4 (fr) 2015-08-21 2019-03-27 Portola Pharmaceuticals, Inc. Modulateurs phénylpipérazine proprotéine convertase subtilisine/kexine de type 9 (pcsk9) et leur utilisation
WO2017034994A1 (fr) 2015-08-21 2017-03-02 Portola Pharmaceuticals, Inc. Composition et procédés d'utilisation de nouveaux petits composés organiques de phénylalanine pour moduler directement l'activité de la protéine pcsk9
EP3337564A4 (fr) 2015-08-21 2019-01-23 Portola Pharmaceuticals, Inc. Composition et procédés d'utilisation de petites molécules de type tétrahydroisoquinoline pour se lier à pcsk9 et moduler l'activité protéique de pcsk9
WO2017147328A1 (fr) 2016-02-23 2017-08-31 Portola Pharmaceuticals, Inc. Composés se liant à la proprotéine convertase subtilisine/kexine de type 9 (pcsk9)
PL3461819T3 (pl) 2017-09-29 2020-11-30 Probiodrug Ag Inhibitory cyklazy glutaminylowej

Family Cites Families (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4839343A (en) * 1987-03-13 1989-06-13 Debiopharm, S.A. Preparation containing hexatriacontapeptides and methods of use
US5026685A (en) * 1988-07-15 1991-06-25 The Salk Institute For Biological Studies NPY peptide analogs
ZA896376B (en) * 1988-08-26 1990-05-30 Merrell Dow Pharma Neuropeptide y agonists

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
See references of WO9500161A1 *

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ZA944338B (en) 1995-02-14
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AU7174494A (en) 1995-01-17
CA2165200A1 (fr) 1995-01-05

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