WO2001038397A1 - N-alkylated peptides having antiangiogenic activity - Google Patents

N-alkylated peptides having antiangiogenic activity Download PDF

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Publication number
WO2001038397A1
WO2001038397A1 PCT/US2000/032105 US0032105W WO0138397A1 WO 2001038397 A1 WO2001038397 A1 WO 2001038397A1 US 0032105 W US0032105 W US 0032105W WO 0138397 A1 WO0138397 A1 WO 0138397A1
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ile
alanyl
sar
arg
ethyl
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PCT/US2000/032105
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English (en)
French (fr)
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Fortuna Haviv
Jack Henkin
Michael F. Bradley
Douglas M. Kalvin
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Abbott Laboratories
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Priority to CA002386893A priority Critical patent/CA2386893A1/en
Priority to JP2001540160A priority patent/JP2003514920A/ja
Priority to BR0010934-7A priority patent/BR0010934A/pt
Priority to EP00980685A priority patent/EP1242455A1/en
Priority to AU17913/01A priority patent/AU1791301A/en
Priority to MXPA02005139A priority patent/MXPA02005139A/es
Publication of WO2001038397A1 publication Critical patent/WO2001038397A1/en
Priority to HK03101124.4A priority patent/HK1050902A1/zh

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K7/00—Peptides having 5 to 20 amino acids in a fully defined sequence; Derivatives thereof
    • C07K7/04—Linear peptides containing only normal peptide links
    • C07K7/08—Linear peptides containing only normal peptide links having 12 to 20 amino acids
    • 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
    • A61P17/00—Drugs for dermatological disorders
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A61P17/06—Antipsoriatics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P19/00—Drugs for skeletal disorders
    • A61P19/02—Drugs for skeletal disorders for joint disorders, e.g. arthritis, arthrosis
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P27/00—Drugs for disorders of the senses
    • A61P27/02—Ophthalmic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/02—Immunomodulators
    • 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
    • A61P9/10—Drugs for disorders of the cardiovascular system for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
    • 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
    • C07K14/78—Connective tissue peptides, e.g. collagen, elastin, laminin, fibronectin, vitronectin or cold insoluble globulin [CIG]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides

Definitions

  • the invention relates to novel compounds having activity useful for treating conditions which arise or are exacerbated by angiogenesis, pharmaceutical compositions comprising the compounds, methods of treatment using the compounds, and methods of inhibiting angiogenesis.
  • Angiogenesis is the fundamental process by which new blood vessels are formed and is essential to a variety of normal body activities (such as reproduction, development " and wound repair). Although the process is not completely understood, it is believed to involve a complex interplay of molecules which both stimulate and inhibit the growth of endothehal cells, the primary cells of the capillary blood vessels. Under normal conditions these molecules appear to maintain the microvasculature in a quiescent state (i.e., one of no capillary growth) for prolonged periods that may last for weeks, or in some cases, decades. However, when necessary, such as during wound repair, these same cells can undergo rapid proliferation and turnover within as little as five days (Folkman, J.
  • angiogenesis is a highly regulated process under normal conditions, many diseases (characterized as "angiogenic diseases") are driven by persistent unregulated angiogenesis. Otherwise stated, unregulated angiogenesis may either cause a particular disease directly or exacerbate an existing pathological condition. For example, ocular neovascularization has been implicated as the most common cause of blindness. In certain existing conditions such as arthritis, newly formed capillary blood vessels invade the joints and destroy cartilage. In diabetes, new capillaries formed in the retina invade the vitreous, bleed, and cause blindness.
  • angiogenesis inhibitors are currently under development for use in treating angiogenic diseases (Gasparini, G. and Harris, A.L., J Clin Oncol 13(3): 765-782, (1995)).
  • a number of disadvantages have been associated with many of these compounds.
  • a potent angiogenesis inhibitor for example suramin, can cause severe systemic toxicity in humans at doses required to reach antitumor activity.
  • Other compounds, such as retinoids, interferons, and antiestrogens are safe for human use, but have only a weak anti- angiogenic effect.
  • the present invention relates to a novel class of compounds having angiogenesis- inhibiting properties.
  • the invention provides nona- and decapeptides wherein the nitrogen atom of at least one of the amide bonds of an amino acid residue in positions 2 through 9 of the peptide is N-alkylated.
  • Compounds of the invention exhibit enhanced metabolic stability, improved pharmacokinetics, increased water solubility, and potentially better oral bioavailability.
  • the present invention provides a compound of formula (I)
  • At least one amide bond of an amino acid residue represented by Xaa 3 , Xaa 4 , Xaa 5 , Xaa 6 , Xaa 7 , Xaa g , Xaa,, and Xaa 10 is N-alkylated;
  • Xaa is absent or Xaa, is selected from the group consisting of hydrogen, N- methylprolyl, and an acyl group, wherein the acyl group is selected from the group consisting of
  • R'-(CH 2 ) n -C(O)- wherein n is an integer from 0 to 8 and R 1 is selected from the group consisting of N-acetylamino, alkoxy, alkyl, aryl, carboxy, cycloalkenyl, cycloalkyl, heterocycle, and hydroxy; and R 2 -CH 2 CH 2 -O-(CH 2 CH 2 O) p -CH 2 -C(O)-, wherein p is an integer from 1 to 8 and R 2 is selected from the group consisting of hydrogen, N-acetylamino, and alkyl; provided that Xaa, is absent only when Xaa 2 is N-(R )-prolyl;
  • Xaa 2 is an N-alkylated amino acid selected from the group consisting of N-(R 3 )- alanyl, N-(R 3 )-glycyl, N-(R 3 )-norvalyl, and N-(R 3 )-prolyl, wherein R 3 is C,-C 5 - alkyl; or Xaa 2 is an N-unalkylated amino acid selected from the group consisting of ⁇ -alanyl,
  • Xaa 3 is an N-alkylated amino acid selected from the group consisting of N-(R 3 )- alanyl, N-(R 3 )-glycyl, N-(R 3 )-leucyl, and N-(R 3 )-phenylalanyl, wherein R 3 is as defined above; or Xaa 3 is an N-unalkylated amino acid selected from the group consisting of alanyl,
  • Xaa 4 is an N-alkylated amino acid selected from the group consisting of N-(R 3 )- alanyl, N-(R 3 )-glycyl, N-(R 3 )-homophenylalanyl, N-(R 3 )-isoleucyl, N-(R 3 )-leucyl,
  • N-(R 3 )-norvalyl N-(R 3 )-phenylalanyl, N-(R 3 )-D-phenylalanyl, N-(R 3 )-seryl, N- (R 3 )-tyrosyl, N-(R 3 )-valyl, and N-(R 3 )-D-valyl, wherein R 3 is as defined above; or Xaa 4 is an N-unalkylated amino acid selected from the group consisting of alanyl, alloisoleucyl, allylglycyl, 2-aminobutyryl,
  • 3-(4-fluorophenyl)alanyl glutaminyl, glycyl, histidyl, homophenylalanyl, homoseryl, isoleucyl, leucyl, lysyl(N-epsilon-acetyl), methionyl, methionyl(sulfone),
  • Xaa 5 is an N-alkylated amino acid selected from the group consisting of N-(R 3 )-D- homophenylalanyl, N-(R 3 )-D-isoleucyl, N-(R 3 )-D-leucyl, and N-(R 3 )-D- phenylalanyl, wherein R 3 is as defined above; or Xaa 5 is an N-unalkylated amino acid selected from the group consisting of
  • Xaa 6 is an N-alkylated amino acid selected from the group consisting of N-(R 3 )- aspartyl, N-(R 3 )-glutamyl, N-(R 3 )-glycyl, N-(R 3 )-seryl, N-(R 3 )-threonyl, N-(R 3 )- threonyl(O-benzyl), and N-(R 3 )-tyrosyl, wherein R 3 is as defined above; or Xaa 6 is an N-unalkylated amino acid selected from the group consisting of alanyl, allothreonyl,
  • D-allothreonyl allylglycyl, asparaginyl, aspartyl, glutaminyl, glycyl, histidyl, homoseryl, D-homoseryl,
  • Xaa 7 is an N-alkylated amino acid selected from the group consisting of N-(R 3 )- alanyl, N-(R 3 )-glycyl, N-(R 3 )-isoleucyl, N-(R 3 )-leucyl, N-(R 3 )-D-leucyl, N-(R 3 )- norleucyl, N-(R 3 )-norvalyl, N-(R 3 )-seryl, N-(R 3 )-threonyl, and N-(R 3 )-valyl, wherein R 3 is as defined above; or Xaa 7 is an N-unalkylated amino acid selected from the group consisting of alanyl, allothreonyl, allylglycyl, 3-(4-amidophenyl)alanyl,
  • D-leucyl lysyl(N-epsilon-acetyl), methionyl sulfone, methionyl sulfoxide, methionyl, norleucyl, norvalyl, D-norvalyl, octylglycyl, ornithyl(N-delta-acetyl), phenylalanyl, propargylglycyl, seryl, D-seryl, threonyl, tryptyl, tyrosyl, and valyl;
  • Xaa 8 is an N-alkylated amino acid selected from the group consisting of N-(R 3 )- alanyl, N-(R 3 )-D-alanyl, N-(R 3 )-isoleucyl, and N-(R 3 )-leucyl, wherein R 3 is as defined above; or Xaa 8 is an N-unalkylated amino acid selected from the group consisting of alanyl, alloisoleucyl,
  • Xaa is the N-alkylated amino acid N-(R 3 )-arginyl, wherein R 3 is as defined above; or Xaa, is an N-unalkylated amino acid selected from the group consisting of
  • Xaa, 0 is an N-alkylated amino acid selected from the group consisting of N-(R 3 )- alanyl, N-(R 3 )-D-alanyl, N-(R 3 )-glycyl, N-(R 3 )-homoalanyl, and N-(R 3 )-norvalyl, wherein R 3 is as defined above; or Xaa
  • Xaa is a hydroxy group or an amino acid amide selected from the group consisting of: alanylamide,
  • D-alanylamide alanylethylamide, D-alanylethylamide, azaglycylamide, glycylamide, glycylethylamide, lysyl(N-epsilon-acetyl), D-lysyl(N-epsilon-acetyl), N-methyl-D-alanylamide, sarcosylamide, serylamide, D-serylamide, a residue represented by the formula
  • R 4 is selected from the group consisting of hydrogen, alkyl, and a 5- to 6-membered cycloalkyl ring;
  • R 5 is selected from the group consisting of hydrogen, alkoxy, alkyl, aryl, cycloalkenyl, cycloalkyl, heterocycle, and hydroxy; provided that s is not zero when R 5 is hydroxy or alkoxy; and
  • R 6 is selected from hydrogen and hydroxy.
  • the present invention provides a method of isolating a receptor from an endothehal cell comprising binding a compound of formula (I) to the receptor to form a peptide receptor complex, isolating the peptide receptor complex, and purifying the receptor.
  • N-acetylamino refers to -NHC(O)CH 3 .
  • acyl refers to an alkyl group attached to the parent molecular moiety through a carbonyl group.
  • alkoxy refers to an alkyl group attached to the parent molecular moiety through an oxygen atom.
  • alkyl refers to a monovalent group derived from a straight or branched chain saturated hydrocarbon by the removal of a hydrogen atom.
  • Preferred alkyl groups for the present invention invention are alkyl groups having from one to five carbon atoms (C,-C 5 alkyl).
  • Alkyl groups of one to three carbon atoms (C,-C 3 alkyl) are more preferred for the present invention.
  • amino refers to -NH 2 .
  • aryl represents a mono- or bicyclic carbocyclic ring system having one or two aromatic rings and is exemplified by phenyl, naphthyl, 1 ,2- dihydronaphthyl, 1,2,3, 4-tetrahy dronaphthyl, fluorenyl, indanyl, indenyl, and the like.
  • the aryl groups of the present invention can be optionally substituted with one, two, three, four, or five substitutents independently selected from the group consisting of alkoxy, alkyl, carboxy, and halo.
  • carbonyl refers to -C(O)-.
  • carboxy refers to CO 2 H.
  • cycloalkenyl refers to a non-aromatic cyclic or bicyclic ring system having three to ten carbon atoms and one to three rings, wherein each five- membered ring has one double bond, each six-membered ring has one or two double bonds, each seven- and eight-membered ring has one to three double bonds, and each nine- to ten-membered ring has one to four double bonds.
  • Examples of cycloalkenyl groups include cyclohexenyl, octahydronaphthalenyl, norbornylenyl, and the like.
  • the cycloalkenyl groups of the present invention can be optionally substituted with one, two, three, four, or five substituents independently selected from the group consisting of alkoxy, alkyl, carboxy, halo, and hydroxy.
  • cycloalkyl refers to a saturated monocyclic, bicyclic, or tricyclic hydrocarbon ring system having three to twelve carbon atoms.
  • examples of cycloalkyl groups include cyclopropyl, cyclopentyl, bicyclo[3.1.1]heptyl, adamantyl, and the like.
  • the cycloalkyl groups of the present invention can be optionally substituted with one, two, three, four, or five substituents independently selected from the group consisting of alkoxy, alkyl, carboxy, halo, and hydroxy.
  • halo refers to F, Cl, Br, or I.
  • heterocycle refers to a five-, six-, or seven-membered ring containing one, two, or three heteroatoms independently selected from the group consisting of nitrogen, oxygen, and sulfur.
  • the five-membered ring has zero to two double bonds and the six- and seven-membered rings have zero to three double bonds.
  • heterocycle also includes bicyclic groups in which the heterocycle ring is fused to an aryl group.
  • the heterocycle groups of the present invention can be attached through a carbon atom or a nitrogen atom in the group.
  • heterocycles include, but are not limited to, furyl, thienyl, pyrrolyl, pyrrolidinyl, oxazolyl, thiazolyl, imidazolyl, imidazolinyl, pyrazolyl, isoxazolyl, isothiazolyl, piperidinyl, morpholinyl, thiomorpholinyl, piperazinyl, pyridinyl, indolyl, indolinyl, benzothienyl, and the like.
  • the heterocycle groups of the present invention can be optionally substituted with one, two, three, or four substituents independently selected from the group consisting of alkoxy, alkyl, carboxy, halo, and hydroxy.
  • hydroxy refers to -OH.
  • nicotinyl refers to the acyl group derived from nicotinic acid, i.e. pyridine-3 -carboxyhc acid.
  • 2-Me-nicotinyl or “2-methylnicotinyl” refers to a nicotinyl moiety substituted with a methyl group at the carbon adjacent to the nitrogen atom in the 2-position.
  • nitrogen protecting group or "N-protecting group,” as used herein, refers to an easily removable group which is known in the art to protect an amino group against undesirable reaction during synthetic procedures and to be selectively removable.
  • nitrogen protecting groups is well known in the art for protecting groups against undesirable reactions during a synthetic procedure and many such protecting groups are known (see, for example, T.H. Greene and P.G.M. Wuts, Protective Groups in Organic Synthesis, 2nd edition, John Wiley & Sons, New York (1991).
  • N- protecting groups include, are not limited to, acyl groups including acetyl, trifluoroacetyl, acylisothiocyanate, aminocaproyl, benzoyl and the like, and acyloxy groups, including t- butyloxycarbonyl (Boc) and carbobenzyloxy (Cbz), 9-fiuorenylmethoxycarbonyl (Fmoc), and the like.
  • esters which hydrolyze in vivo and include those that break down readily in the human body to leave the parent compound or a salt thereof.
  • Suitable ester groups include, for example, those derived from pharmaceutically acceptable aliphatic carboxyhc acids, particularly alkanoic, alkenoic, cycloalkanoic and alkanedioic acids, in which each alkyl or alkenyl moiety advantageously has not more than six carbon atoms.
  • esters include formates, acetates, propionates, butyrates, acrylates and ethylsuccinates.
  • prodrugs refers to those prodrugs of the compounds of the present invention which are, within the scope of sound medical judgement, suitable for use in contact with the tissues of humans and lower animals with undue toxicity, irritation, allergic response, and the like, commensurate with a reasonable benefit/risk ratio, and effective for their intended use, as well as the zwitterionic forms, where possible, of the compounds of the invention.
  • prodrug refers to compounds that are rapidly transformed in vivo to yield the parent compound of the above formula, for example by hydrolysis in blood.
  • salts or zwitterionic forms of the compounds of the present invention which are water or oil- soluble or dispersible, which are suitable for treatment of diseases without undue toxicity, irritation, and allergic response; which are commensurate with a reasonable benefit/risk ratio, and which are effective for their intended use.
  • the salts can be prepared during the final isolation and purification of the compounds or separately by reacting an amino group with a suitable acid.
  • Representative acid addition salts include acetate, adipate, alginate, citrate, aspartate, benzoate, benzenesulfonate, bisulfate, butyrate, camphorate, camphorsulfonate, digluconate, glycerophosphate, hemisulfate, heptanoate, hexanoate, formate, fumarate, hydrochloride, hydrobromide, hydroiodide, 2-hydroxyethansulfonate (isethionate), lactate, maleate, mesitylenesulfonate, methanesulfonate, naphthylenesulfonate, nicotinate, 2-naphthalenesulfonate, oxalate, pamoate, pectinate, persulfate, 3-phenylproprionate, picrate, pivalate, propionate, succinate, tartrate, trichloroacetate,trifluoroacetate, phosphate,
  • amino groups in the compounds of the present invention can be quaternized with methyl, ethyl, propyl, and butyl chlorides, bromides, and iodides; dimethyl, diethyl, dibutyl, and diamyl sulfates; decyl, lauryl, myristyl, and steryl chlorides, bromides, and iodides; and benzyl and phenethyl bromides.
  • acids which can be employed to form therapeutically acceptable addition salts include inorganic acids such as hydrochloric, hydrobromic, sulfuric, and phosphoric, and organic acids such as oxalic, maleic, succinic, and citric.
  • Basic addition salts can be prepared during the final isolation and purification of the compounds by reacting a carboxy group with a suitable base such as the hydroxide, carbonate, or bicarbonate of a metal cation or with ammonia or an organic primary, secondary, or tertiary amine.
  • a suitable base such as the hydroxide, carbonate, or bicarbonate of a metal cation or with ammonia or an organic primary, secondary, or tertiary amine.
  • the cations of pharmaceutically acceptable salts include lithium, sodium, potassium, calcium, magnesium, and aluminum, as well as nontoxic quaternary amine cations such as ammonium, tetramethylammonium, tetraethylammonium, methylamine, dimethylamine, trimethylamine, triethylamine, diethylamine, ethylamine, tributylamine, pyridine, N,N-dimethylaniline, N- methylpiperidine, N-methylmorpholine, dicyclohexylamine, procaine, dibenzylamine, N,N-dibenzylphenethylamine, 1 -ephenamine, and N,N'-dibenzylethylenediamine.
  • nontoxic quaternary amine cations such as ammonium, tetramethylammonium, tetraethylammonium, methylamine, dimethylamine, trimethylamine, triethylamine, diethyl
  • compositions useful for the formation of base addition salts include ethylenediamine, ethanolamine, diethanolamine, piperidine, and piperazine.
  • pharmaceutically acceptable solvate refers to an aggregate that comprises one or more molecules of the solute, such as a compound of formula (I), with one or more molecules of solvent.
  • receptor refers to chemical groups or molecules on the cell surface or in the cell interior that have an affinity for a specific chemical group or molecule. Isolation of receptors relevant to the antiangiogenic activity of the peptide of the invention can provide useful diagnostic tools.
  • shikimyl refers to the acyl residue derived from shikimic acid or [3R-(3 ⁇ ,4 ⁇ ,5 ⁇ )-3,4,5-trihydroxy]-l-cyclohexene-l-carboxylic acid.
  • a "dihydroshikimyl” group denotes the fully saturated analog of shikimic acid.
  • succinyl refers to the acyl residue derived from succinic acid or (l,4-dioxobutyl)-l -carboxyhc acid. Unless indicated otherwise by a "D-" prefix, e.g.
  • ⁇ -N-terminal refers to the free ⁇ -amino group of an amino acid in a peptide
  • ⁇ -C-terminal refers to the free ⁇ -carboxylic acid terminus of an amino acid in a peptide
  • the present invention relates to compounds of formula (I), wherein Xaa 2 -Xaa, 0 each represent an amino acyl residue; Xaa, may be absent or Xaa, is hydrogen, N-methylprolyl, or an acyl group; and Xaa,, is a hydroxy group, an amino acid amide, or an amino residue.
  • the amino acyl residues represented by Xaa 2 - Xaa, 0 can have an N- alkylated or an N-unalkylated amide bond.
  • At least one of the amide bonds on a residue represented by Xaa 3 , Xaa 4 , Xaa 5 , Xaa 6 , Xaa 7 , Xaa 8 , Xaa,, or Xaa, 0 is N-alkylated.
  • Xaa is absent or is selected from the group consisting of hydrogen; N- methylprolyl; R'-(CH 2 ) n -C(O)-, wherein n is an integer from 0 to 8 and R 1 is selected from the group consisting of N-acetylamino, alkoxy, alkyl, aryl, carboxy, cycloalkenyl, cycloalkyl, heterocycle, and hydroxy; and R 2 -CH 2 CH 2 -O-(CH 2 CH 2 O) -CH 2 -C(O)-, wherein p is an integer from 1 to 8 and R 2 is selected from the group consisting of hydrogen, N-acetylamino, and alkyl.
  • Xaa is absent or is selected from the group consisting of acetyl, N-methylprolyl, and succinyl.
  • Amino acyl residues suitable for the Xaa 2 position include N-methylalanyl, sarcosyl, N-ethylglycyl, N-methylnorvalyl, N-methylprolyl, ⁇ -alanyl, D-alanyl, 4- aminobutyryl, (lR,3S)-l-aminocyclopentane-3-carbonyl, (lS,3R)-l-aminocyclopentane-3- carbonyl, ( 1 R,4S)- 1 -aminocyclopent-2-ene-4-carbonyl, ( 1 S,4R)- 1 -aminocyclopent-2-ene- 4-carbonyl, asparaginyl, 3-(4-chlorophenyl)alanyl, 3-(4-cyanophenyl)alanyl, glutaminyl, glutamyl, glycyl, 4-hydroxyprolyl, 3-(4-methylphenyl)alanyl, pro
  • Suitable amino acyl residues for Xaa 3 include N-methylalanyl, sarcosyl, N- methylleucyl, N-methylphenylalanyl, alanyl, (lS,3R)-l-aminocyclopentane-3-carbonyl, (lS,4R)-l-aminocyclopent-2-ene-4-carbonyl, asparaginyl, aspartyl, 3-(3- cyanophenyl)alanyl, 3-(4-cyanophenyl)alanyl, glutaminyl, glycyl, leucyl, lysyl(N-epsilon- acetyl), 3-(4-methylphenyl)alanyl, norvalyl, prolyl, and phenylalanyl.
  • the preferred amino acid residues for Xaa 3 are N-methylalanyl and glycyl.
  • N-alkylated residues suitable for Xaa 4 include N-methylalanyl, sarcosyl, N- methylhomophenylalanyl, N-methylisoleucyl, N-methylleucyl, N-methylnorvalyl, N- methylphenylalanyl, N-methyl-D-phenylalanyl, N-methylseryl, N-methyltyrosyl, N- methylvalyl, and N-methyl-D-valyl.
  • N-Unalkylated amino acyl residues suitable for Xaa 4 are alanyl, alloisoleucyl, allylglycyl, 2-aminobutyryl, (lR,4S)-aminocyclopent-2-ene-4- carbonyl, asparaginyl, aspartyl, 3-[2-(5-bromothienyl)]alanyl, 3-(3-chlorophenyl)alanyl, 3- (4-chlorophenylalanyl), 3-(3-cyanophenyl)alanyl, cyclohexylalanyl, 3 -(3 ,4- dimethoxyphenyl)alanyl, 3-(3-_luorophenyl)alanyl, 3-(4-fluorophenylalanyl), glutaminyl, glycyl, histidyl, homophenylalanyl, homoseryl, isoleucyl, leuc
  • Preferred amino acyl residues for Xaa 4 are N-methylalanyl, N-methylisoleucyl, N-methylleucyl, N-methylnorvalyl, N- methylphenylalanyl, N-methyl-D-phenylalanyl, N-methylvalyl, N-methyl-D-valyl, asparaginyl, glutaminyl, isoleucyl, phenylalanyl, and valyl.
  • N-alkylated amino acyl residues suitable for Xaa 5 are N-methyl-D- homophenylalanyl, N-methyl-D-isoleucyl, N-methyl-D-leucyl, and N-(R 3 )-D- phenylalanyl.
  • N-unalkylated amino acyl residues suitable for Xaa 5 are D-alanyl, alloisoleucyl, D-alloisoleucyl, D-2-aminobutyryl, D-3-(4-aminophenyl)alanyl, D- asparaginyl, D-3-(3-benzothienyl)alanyl, D-t-butylglycyl, D-(4-chlorophenyl)alanyl, D- citrullyl, D-3-(3-cyanophenyl)alanyl, D-cyclohexylalanyl, cyclohexylglycyl, D- cysteinyl(S-acetamidomethyl), D-cysteinyl(S-t-butyl), D-3-(3,4-difluorophenyl)alanyl, D- (3,4-dimethoxyphenyl)alanyl, D-glutaminyl
  • amino acyl residues preferred for Xaa 5 are N-methyl-D-leucyl, D-alloisoleucyl, D- isoleucyl, D-leucyl, D-homophenylalanyl, and D-penacillaminyl(S-methyl).
  • N-alkylated amino acids suitable for Xaa 6 are N-methylaspartyl, N- methylglutamyl, sarcosyl, N-methylseryl, N-methylthreonyl, N-methylthreonyl(O-benzyl), and N-methyltyrosyl.
  • N-unalkylated amino acyl residues suitable for Xaa 6 are alanyl, allothreonyl, D-allothreonyl, allylglycyl, glutaminyl, glycyl, histidyl, homoseryl, D- homoseryl, 3-(4-hydroxymethylphenyl)alanyl, isoleucyl, lysyl(N-epsilon-acetyl), methionyl, 3-(naphth-2-yl)alanyl, norvalyl, octylglycyl, prolyl, 3-(3-pyridyl)alanyl, seryl, D-seryl, threonyl, D-threonyl, tryptyl, tyrosyl, and tyrosyl(O-methyl).
  • the preferred amino acyl residues for Xaa ⁇ are N-methylaspartyl, N-methylglutamyl, sarcosyl, N- methylseryl, N-methyltyrosyl, N-methylthreonyl(O-benzyl), allothreonyl, seryl, threonyl, and tyrosyl.
  • N-Alkylated amino acyl residues suitable for Xaa 7 are N-methylalanyl, sarcosyl, N- methylisoleucyl, N-methyleucyl, N-methyl-D-leucyl, N-methylnorleucyl, N- methylnorvalyl, N-methylseryl, N-methylthreonyl, and N-methyl valyl.
  • N-unalkylated amino acyl residues suitable for Xaa 7 are alanyl, allothreonyl, allylglycyl, 3-(4- amidophenyl)alanyl, 2-aminobutyryl, arginyl, asparaginyl, cyclohexylalanyl, glutaminyl, D-glutaminyl, glycyl, homoalanyl, homoseryl, 4-hydroxyprolyl, leucyl, D-leucyl, lysyl(N- epsilon-acetyl), methionyl, methionyl sulfone, methionyl sulfoxide, norleucyl, norvalyl, D- norvalyl, octylglycyl, ornithyl(N-delta-acetyl), phenylalanyl, propargylglycyl, se
  • the amino acyl residue for Xaa 7 is selected from the group consisting of N-methylalanyl, sarcosyl, N-methylisoleucyl, N- methylleucyl, N-methyl-D-leucyl, N-methylnorleucyl, N-methylnorvalyl, N-methylseryl, N-methylthreonyl, N-methylvalyl, norleucyl, norvalyl, and seryl.
  • Suitable N-alkyl amino acyl residues for Xaa 8 include N-methylalanyl, N-methyl- D-alanyl, N-methylisoleucyl, and N-methylleucyl.
  • N-Unalkylated amino acyl residues suitable for Xaa 8 include alanyl, alloisoleucyl, D-alloisoleucyl, allylglycyl, citrullyl, glycyl, isoleucyl, D-isoleucyl, leucyl, D-leucyl, lysyl(N-epsilon-acetyl), D-lysyl(N- epsilon-acetyl), methionyl, 3-(naphth-l-yl)alanyl, norvalyl, prolyl, D-prolyl, and valyl.
  • the amino acyl residue for Xaa 8 is selected from the group consisting of N- methylalanyl, N-methyl-D-alanyl, N-methylisoleucyl, N-methylleucyl, isoleucyl, D- isoleucyl, and D-lysyl(N-epsilon-acetyl).
  • N-alkylated amino acyl residues suitable for Xaa is N-methylarginyl.
  • N-Unalkylated amino acyl residues for Xaa are selected from the group consisting of [(4- amino-N-isopropyl)cyclohexyl]alanyl, 3-(4-amino-N-isopropylphenyl)alanyl, arginyl, arginyl(N vf diethyl), citrullyl, glutaminyl, 3-(4-guanidinophenyl)alanyl, histidyl, homoarginyl, lysyl(N-epsilon-isopropyl), lysyl(N-epsilon-nicotinyl), lysyl, norarginyl, ornithyl, ornithyl(N-delta-imidazolinyl), ornithyl(N-methyl
  • Preferred amino acyl residues for Xaa are arginyl and N-methylarginyl.
  • N-Alkylated amino acids suitable for the Xaa, 0 position include N-methylalanyl, N-methyl-D-alanyl, sarcosyl, N-methylhomoalanyl, and N-methylnorvalyl.
  • residues suitable for Xaa 10 include D-alanyl, 2-aminoburyryl, D-2-aminobutyryl, 2- aminoisobutyryl, 3,4-dehydroprolyl, 4-hydroxyprolyl, phenylalanyl, prolyl, D-prolyl, l,2,3,4-tetrahydroisoquinoline-3-carbonyl, and D-valyl.
  • the amino acyl residues preferred for Xaa, 0 are N-methylalanyl, sarcosyl, N-methylnorvalyl, and prolyl.
  • one or two residues selected from Xaa 3 , Xaa 4 , Xaa 5 , Xaa 6 , Xaa 7 , Xaa 8 , Xaa, and Xaa, 0 has an N-alkylated amino acyl residue.
  • the more preferred compounds of the invention have one N-alkylated amide bond on an amino acyl residue not including Xaa,, as represented by Xaa 3 , Xaa 4 , Xaa 5 , Xaa 6 , Xaa 7 , Xaa 8 , Xaa, or Xaa, 0
  • Xaa is a hydroxy group or an amino acid amide selected from the group consisting of alanylamide, D-alanylamide, alanylethylamide, D-alanylethylamide, azaglycylamide, glycylamide, glycylethylamide, lysyl(N-epsilon-acetyl), D-lysyl(N- epsilon-acetyl), N-methyl-D-alanylamide, sarcosylamide, serylamide, and D-serylamide; or Xaa, is a group represented by the formula
  • the preferred Xaa n groups for modifying the C-terminus of the invention are NH-ethyl and D-alanylamide.
  • compositions The compounds of the invention, including not limited to those specified in the examples, possess anti-angiogenic activity.
  • angiogenesis inhibitors such compounds are useful in the treatment of both primary and metastatic solid tumors, including carcinomas of breast, colon, rectum, lung, oropharynx, hypopharynx, esophagus, stomach, pancreas, liver, gallbladder and bile ducts, small intestine, urinary tract (including kidney, bladder and urothelium), female genital tract (including cervix, uterus, and ovaries as well as choriocarcinoma and gestational trophoblastic disease), male genital tract (including prostate, seminal vesicles, testes and germ cell tumors), endocrine glands (including the thyroid, adrenal, and pituitary glands), and skin, as well as hemangiomas, melanomas, sarcomas (including those arising from bone and soft tissues as well as Kaposi's
  • Such compounds may also be useful in treating solid tumors arising from hematopoietic malignancies such as leukemias (i.e. chloromas, plasmacytomas and the plaques and tumors of mycosis fungosides and cutaneous T-cell lymphoma/leukemia) as well as in the treatment of lymphomas (both Hodgkin' s and non-Hodgkin' s lymphomas).
  • leukemias i.e. chloromas, plasmacytomas and the plaques and tumors of mycosis fungosides and cutaneous T-cell lymphoma/leukemia
  • lymphomas both Hodgkin' s and non-Hodgkin' s lymphomas.
  • these compounds may be useful in the prevention of metastases from the tumors described above either when used alone or in combination with radiotherapy and/or other chemo therapeutic agents.
  • autoimmune diseases such as rheumatoid, immune and degenerative arthritis
  • various ocular diseases such as diabetic retinopathy, retinopathy of prematurity, corneal graft rejection, retrolental fibroplasia, neovascular glaucoma, rubeosis, retinal neovascularization due to macular degeneration, hypoxia, angiogenesis in the eye associated with infection or surgical intervention, and other abnormal neovascularization conditions of the eye
  • skin diseases such as psoriasis
  • blood vessel diseases such as hemagiomas, and capillary proliferation within atherosclerotic plaques
  • Osier- Webber Syndrome myocardial angiogenesis
  • plaque neovascularization telangiectasia
  • hemophiliac joints angiofibroma
  • wound granulation such as rheumatoid, immune and degenerative arthritis
  • various ocular diseases such as diabetic retinopathy, retinopathy of prematurity
  • Other uses include the treatment of diseases characterized by excessive or abnormal stimulation of endothehal cells, including not limited to intestinal adhesions, Crohn's disease, atherosclerosis, scleroderma, and hypertrophic scars, i.e. keloids.
  • Another use is as a birth control agent, by inhibiting ovulation and establishment of the placenta.
  • the compounds of the invention are also useful in the treatment of diseases that have angiogenesis as a pathologic consequence such as cat scratch disease (Rochele minutesalia quintosa) and ulcers (Helicobacter pylori).
  • the compounds of the invention are also useful to reduce bleeding by administration prior to surgery, especially for the treatment of resectable tumors.
  • the compounds of the invention may be used in combination with other compositions and procedures for the treatment of diseases.
  • a tumor may be treated conventionally with surgery, radiation or chemotherapy combined with a peptide of the present invention and then a peptide of the present invention may be subsequently administered to the patient to extend the dormancy of micrometastases and to stabilize and inhibit the growth of any residual primary tumor.
  • the compounds of the invention may be combined with pharmaceutically acceptable excipients, and optionally sustained-release matrices, such as biodegradable polymers, to form therapeutic compositions.
  • a sustained-release matrix is a matrix made of materials, usually polymers, which are degradable by enzymatic or acid-base hydrolysis or by dissolution.
  • a sustained-release matrix desirably is chosen from biocompatible materials such as liposomes, polylactides (polylactic acid), polyglycolide (polymer of glycolic acid), polylactide co-glycolide (copolymers of lactic acid and glycolic acid) polyanhydrides, poly(ortho)esters, polypeptides, hyaluronic acid, collagen, chondroitin sulfate, carboxyhc acids, fatty acids, phospholipids, polysaccharides, nucleic acids, polyamino acids, amino acids such as phenylalanine, tyrosine, isoleucine, polynucleotides, polyvinyl propylene, polyvinylpyrrolidone and silicone.
  • a preferred biodegradable matrix is a matrix of one of either polylactide, polyglycolide, or polylactide co-glycolide (co-poly
  • a therapeutically effective amount of one of the compounds of the present invention may be employed in pure form or, where such forms exist, in pharmaceutically acceptable salt form.
  • a “therapeutically effective amount” of the compound of the invention is meant a sufficient amount of the compound to treat an angiogenic disease, (for example, to limit tumor growth or to slow or block tumor metastasis) at a reasonable benefit/risk ratio applicable to any medical treatment. It will be understood, however, that the total daily usage of the compounds and compositions of the present invention will be decided by the attending physician within the scope of sound medical judgment.
  • the specific therapeutically effective dose level for any particular patient will depend upon a variety of factors including the disorder being treated and the severity of the disorder; activity of the specific compound employed; the specific composition employed, the age, body weight, general health, sex and diet of the patient; the time of administration, route of administration, and rate of excretion of the specific compound employed; the duration of the treatment; drugs used in combination or coincidential with the specific compound employed; and like factors well known in the medical arts. For example, it is well within the skill of the art to start doses of the compound at levels lower than those required to achieve the desired therapeutic effect and to gradually increase the dosage until the desired effect is achieved.
  • a compound of the present invention may be administered as pharmaceutical compositions containing the compound of interest in combination with one or more pharmaceutically acceptable excipients.
  • a pharmaceutically acceptable carrier or excipient refers to a non-toxic solid, semi-solid or liquid filler, diluent, encapsulating material or formulation auxiliary of any type.
  • the compositions may be administered parenterally, intracisternally, intravaginally, intraperitoneally, topically (as by powders, ointments, drops or transdermal patch), rectally, or bucally.
  • parenteral refers to modes of administration which include intravenous, intramuscular, intraperitoneal, intrasternal, subcutaneous and intraarticular injection and infusion.
  • compositions for parenteral injection comprise pharmaceutically- acceptable sterile aqueous or nonaqueous solutions, dispersions, suspensions or emulsions, as well as sterile powders for reconstitution into sterile injectable solutions or dispersions just prior to use.
  • suitable aqueous and nonaqueous carriers, diluents, solvents or vehicles include water, ethanol, polyols (such as glycerol, propylene glycol, polyethylene glycol, and the like), carboxymethylcellulose and suitable mixtures thereof, vegetable oils (such as olive oil), and injectable organic esters such as ethyl oleate.
  • Proper fluidity may be maintained, for example, by the use of coating materials such as lecithin, by the maintenance of the required particle size in the case of dispersions, and by the use of surfactants.
  • compositions may also contain adjuvants such as preservative, wetting agents, emulsifying agents, and dispersing agents. Prevention of the action of microorganisms may be ensured by the inclusion of various antibacterial and antifungal agents, for example, paraben, chlorobutanol, phenol sorbic acid, and the like. It may also be desirable to include isotonic agents such as sugars, sodium chloride, and the like. Prolonged absorption of the injectable pharmaceutical form may be brought about by the inclusion of agents which delay absorption, such as aluminum monostearate and gelatin.
  • Injectable depot forms are made by forming microencapsule matrices of the drug in biodegradable polymers such as polylactide-polyglycolide, poly(orthoesters), poly(anhydrides), and (poly)glycols, such as PEG. Depending upon the ratio of drug to polymer and the nature of the particular polymer employed, the rate of drug release can be controlled. Depot injectable formulations are also prepared by entrapping the drug in liposomes or microemulsions which are compatible with body tissues.
  • the injectable formulations may be sterilized, for example, by filtration through a bacterial-retaining filter, or by incorporating sterilizing agents in the form of sterile solid compositions which can be dissolved or dispersed in sterile water or other sterile injectable medium just prior to use.
  • Topical administration includes administration to the skin or mucosa, including surfaces of the lung and eye.
  • Compositions for topical administration may be prepared as a dry powder which may be pressurized or non-pressurized.
  • the active ingredient in finely divided form may be used in admixture with a larger-sized pharmaceutically-acceptable inert carrier comprising particles having a size, for example, of up to 100 micrometers in diameter.
  • suitable inert carriers include sugars such as lactose.
  • at least 95% by weight of the particles of the active ingredient have an effective particle size in the range of 0.01 to 10 micrometers.
  • the composition may be pressurized and contain a compressed gas, such as nitrogen or a liquified gas propellant.
  • a compressed gas such as nitrogen or a liquified gas propellant.
  • the liquified propellant medium and indeed the total composition is preferably such that the active ingredient does not dissolve therein to any substantial extent.
  • the pressurized composition may also contain a surface active agent, such as a liquid or solid non-ionic surface active agent or may be a solid anionic surface active agent. It is preferred to use the solid anionic surface active agent in the form of a sodium salt.
  • a further form of topical administration is to the eye.
  • a compound of the invention is delivered in a pharmaceutically acceptable ophthalmic vehicle, such that the compound is maintained in contact with the ocular surface for a sufficient time period to allow the compound to penetrate the corneal and internal regions of the eye, as for example the anterior chamber, posterior chamber, vitreous body, aqueous humor, vitreous humor, cornea, iris/ciliary, lens, choroid retina and sclera.
  • the pharmaceutically- acceptable ophthalmic vehicle may, for example, be an ointment, vegetable oil or an encapsulating material.
  • the compounds of the invention may be injected directly into the vitreous and aqueous humour.
  • compositions for rectal or vaginal administration are preferably suppositories which may be prepared by mixing the compounds of this invention with suitable non- irritating excipients or carriers such as cocoa butter, polyethylene glycol or a suppository wax which are solid at room temperature liquid at body temperature and therefore melt in the rectum or vaginal cavity and release the active compound.
  • suitable non- irritating excipients or carriers such as cocoa butter, polyethylene glycol or a suppository wax which are solid at room temperature liquid at body temperature and therefore melt in the rectum or vaginal cavity and release the active compound.
  • Liposomes are generally derived from phospholipids or other lipid substances. Liposomes are formed by mono- or multi-lamellar hydrated liquid crystals that are dispersed in an aqueous medium. Any non-toxic, physiologically- acceptable and metabolizable lipid capable of forming liposomes can be used.
  • the present compositions in liposome form can contain, in addition to a compound of the present invention, stabilizers, preservatives, excipients, and the like.
  • the preferred lipids are the phospholipids and the phosphatidyl cholines (lecithins), both natural and synthetic.
  • the compounds of the invention can be administered as the sole active pharmaceutical agent, they may also be used in combination with one or more agents which are conventionally administered to patients for treating angiogenic diseases.
  • the compounds of the invention are effective over the short term to make tumors more sensitive to traditional cytotoxic therapies such as chemicals and radiation.
  • the compounds of the invention also enhance the effectiveness of existing cytotoxic adjuvant anti-cancer therapies.
  • the compounds of the invention may also be combined with other antiangiogenic agents to enhance their effectiveness, or combined with other antiangiogenic agents and administered togdiethyl ether with other cytotoxic agents.
  • compounds of the invention when used in the treatment of solid tumors, compounds of the invention may be administered with IL-12, retinoids, interferons, angiostatin, endostatin, thalidomide, thrombospondin-1, thrombospondin-2, captopryl, angioinhibins, TNP-470, pentosan polysulfate, platelet factor 4, LM-609, SU-5416, CM-101, Tecogalan, plasminogen-K-5, vasostatin, vitaxin, vasculostatin, squalamine, marimastat or other MMP inhibitors, anti- neoplastic agents such as alpha inteferon, COMP (cyclophosphamide, vincristine, methotrexate and prednisone),
  • Total daily dose of the compositions of the invention to be administered to a human or other mammal host in single or divided doses may be in amounts, for example, from 0.0001 to 300 mg/kg body weight daily and more usually 1 to 300 mg/kg body weight.
  • agents which can be combined with the compound of the present invention for the inhibition, treatment or prophylaxis of angiogenic diseases are not limited to those listed above, include in principle any agents useful for the treatment or prophylaxis of angiogenic diseases.
  • the peptides of the invention may be used for the development of affinity columns for isolation of receptors relevant to the antiangiogenic activity of the peptide of the invention, e.g. TSP-1 receptor, in, for example, cultured endothehal cells.
  • isolation and purification of the receptor may be followed by amino acid sequencing to identify and isolate polynucleotides which encode the receptor. Recombinant expression of this receptor would allow greater amounts of receptor to be produced, e.g. to produce a sufficient quantity for use in high throughput screening assays to identify other angiogenesis inhibitors.
  • HMVEC human microvascular endothehal
  • the HMVEC migration assay was carried out using Human Microvascular Endothehal Cells-Dermal (single donor) and Human Microvascular Endothehal Cells, (neonatal).
  • the BCE or HMVEC cells were starved overnight in DME containing 0.1% bovine serum albumin (BSA). Cells were then harvested with trypsin and resuspended in DME with 0.1% BSA at a concentration of 1.5 X 10 ⁇ cells per mL. Cells were added to the bottom of a 48 well modified Boy den chamber (Nucleopore Corporation, Cabin John, MD).
  • the chamber was assembled and inverted, and cells were allowed to attach for 2 hours at 37 °C to polycarbonate chemotaxis membranes (5 ⁇ m pore size) that had been soaked in 0.1 % gelatin overnight and dried.
  • the chamber was then reinverted, and test substances (total volume of 50 ⁇ L), including activators, 15 ng/mL bFGF/VEGF, were added to the wells of the upper chamber.
  • the apparatus was incubated for 4 hours at 37 °C. Membranes were recovered, fixed and stained (Diff Quick, Fisher Scientific) and the number of cells that had migrated to the upper chamber per 3 high power fields counted.
  • polypeptides of the present invention may be synthesized by many techniques that are known to those skilled in the art.
  • solid phase peptide synthesis a summary of the many techniques may be found in J.M. Stewart and J.D. Young, Solid Phase Peptide Synthesis, W.H. Freeman Co. (San Francisco), 1963 and J. Meienhofer, Hormonal Proteins and Peptides, vol. 2, p. 46, Academic Press (New York), 1973.
  • For classical solution synthesis see G. Schroder and K. Lupke, The Peptides, vol. 1, Academic Press (New York), 1965.
  • Reagents, resins, amino acids, and amino acid derivatives are commercially available and can be purchased from Chem-Impex International, Inc. (Wood Dale, IL, U.S.A.) or Calbiochem-Novabiochem Corp. (San Diego, CA, U.S.A.) unless otherwise noted herein.
  • these methods comprise the sequential addition of one or more amino acids or suitably protected amino acids to a growing peptide chain. Normally, either the amino or carboxy group of the first amino acid is protected by a suitable protecting group.
  • the protected or derivatized amino acid can then be either attached to an inert solid support or utilized in solution by adding the next amino acid in the sequence having the complimentary (amino or carboxy) group suitably protected, under conditions suitable for forming the amide linkage.
  • the protecting group is then removed from this newly added amino acid residue and the next amino acid (suitably protected) is then added, and so forth. After all the desired amino acids have been linked in the proper sequence, any remaining protecting groups (and any solid support) are removed sequentially or concurrently, to afford the final polypeptide.
  • a particularly preferred method of preparing compounds of the present invention involves solid phase peptide synthesis.
  • the ⁇ -amino function is protected by an acid or base sensitive group.
  • Such protecting groups should have the properties of being stable to the conditions of peptide linkage formation, while being readily removable without destruction of the growing peptide chain or racemization of any of the chiral centers contained therein.
  • Suitable protecting groups are 9-fluorenylmethyloxycarbonyl (Fmoc), t-butoxycarbonyl (Boc), benzyloxycarbonyl (Cbz), biphenylisopropyl-oxycarbonyl, t- amyloxycarbonyl, isobornyloxycarbonyl, ( ⁇ , ⁇ )-dimethyl-3,5- dimethoxybenzyloxycarbonyl, O-nitrophenylsulfenyl, 2-cyano-t-butyloxycarbonyl, and the like.
  • the 9-fluorenylmethyloxycarbonyl (Fmoc) protecting group is preferred.
  • Particularly preferred side chain protecting groups are: for arginine and lysine: acetyl (Ac), adamantyloxycarbonyl, benzyloxycarbonyl (Cbz), t-butyloxycarbonyl (Boc), 4-methoxybenzenesulfonyl, N G -4-methoxy-2,3,6-trimethylbenzenesulfonyl (Mtr), nitro, 2,2,5,7,8-pentamethylchroman-6-sulfonyl (Pmc), and p-toluenesulfonyl; for asparagine: trityl (Trt); for aspartyl: t-butyl (t-Bu); for glutamyl: t-butyl (t-Bu); for glutaminyl: trityl (Trt); for histidine: trityl (Trt), benzyl, benzyloxycarbonyl (Cbz
  • the C-terminal amino acid is attached to a suitable solid support or resin.
  • suitable solid supports useful for the above synthesis are those materials which are inert to the reagents and reaction conditions of the stepwise condensation-deprotection reactions, as well as being insoluble in the media used.
  • the preferred solid support for synthesis of C-terminal carboxy peptides is 4-hydroxymethyl- phenoxymethyl-copoly(styrene-l% divinylbenzene).
  • the preferred solid support for C- terminal amide peptides is 4-(2',4'-dimethoxyphenyl-Fmoc-aminomethyl)phenoxy- acetamidoethyl resin available from Applied Biosystems.
  • the C-terminal amino acid is coupled to the resin by means of a coupling mediated by N,N'-dicyclohexylcarbodiimide (DCC), N,N'-diisopropylcarbodiimide (DIC), [O-(7- azabenzotriazol-l-yl)-l,l,3,3-tetramethyluronium hexafluorophoshpate] (HATU), or O- benzotriazol-l-yl-N,N,N',N'-tetramethyluroniumhexafluorophosphate (HBTU), with or without 4-dimethylaminopyridine (DMAP), 1 -hydroxybenzotriazole (HOBT), benzotriazol-1 -yloxy-tris(dimethylamino)phosphoniumhexafluorophosphate (BOP) or bis(2-oxo-3-oxazolidinyl)phosphine chloride (BOPCl), for about 1
  • the Fmoc group is cleaved with a secondary amine, preferably piperidine, prior to coupling with the C-terminal amino acid as described above.
  • the preferred reagents used in the coupling to the deprotected 4-(2',4'-dimethoxyphenyl- Fmoc-aminomethyl)phenoxyacetamidoethyl resin are O-benzotriazol-l-yl-N,N,N',N'- tetramethyluroniumhexafluorophosphate (HBTU, 1 equiv.) and 1 -hydroxybenzotriazole (HOBT, 1 equiv.), or [O-(7-azabenzotriazol-l-yl)-l,l,3,3-tetramethyluronium hexafluorophoshpate] (HATU, 1 equiv.), in DMF.
  • the coupling of successive protected amino acids can be carried out in an automatic polypeptide synthesizer as is well known in the art.
  • the ⁇ -amino function in the amino acids of the growing peptide chain are protected with Fmoc.
  • the removal of the Fmoc protecting group from the N-terminal side of the growing peptide is accomplished by treatment with a secondary amine, preferably piperidine.
  • Each protected amino acid is then introduced in about 3-fold molar excess and the coupling is preferably carried out in DMF.
  • the coupling agent is normally O-benzotriazol-1-yl- N,N,N',N'-tetramethyluroniumhexafluorophosphate (HBTU, 1 equiv.) and 1 -hydroxybenzotriazole (HOBT, 1 equiv.) or [O-(7-azabenzotriazol-l-yl)-l, 1,3,3- tetramethyluronium hexafluorophoshpate] (HATU, 1 equiv.).
  • the polypeptide is removed from the resin and deprotected, either in succession or in a single operation. Removal of the polypeptide and deprotection can be accomplished in a single operation by treating the resin-bound polypeptide with a cleavage reagent, for example trifluoroacetic acid containing thianisole, water, or ethanedithiol.
  • a cleavage reagent for example trifluoroacetic acid containing thianisole, water, or ethanedithiol.
  • the resin is cleaved by aminolysis with an alkylamine.
  • the peptide may be removed by transesterification, e.g. with methanol, followed by aminolysis or by direct transamidation.
  • the protected peptide may be purified at this point or taken to the next step directly.
  • the removal of the side chain protecting groups is accomplished using the cleavage cocktail described above.
  • the fully deprotected peptide is purified by a sequence of chromatographic steps employing any or all of the following types: ion exchange on a weakly basic resin in the acetate form; hydrophobic adsorption chromatography on underivitized polystyrene- divinylbenzene (for example, AMBERLITE® XAD); silica gel adsorption chromatography; ion exchange chromatography on carboxymethylcellulose; partition chromatography, e.g. on SEPHADEX® G-25, LH-20 or countercurrent distribution; high performance liquid chromatography (HPLC), especially reverse-phase HPLC on octyl- or octadecylsilyl-silica bonded phase column packing.
  • HPLC high performance liquid chromatography
  • NMP N- methylpyrrolidinone
  • HATU [O-(7-azabenzotriazol-l-yl)-l,l,3,3-tetramethyluronium hexafluorophosphate]
  • DMF N,N-dimethylformamide
  • TFA trifluoroacetic acid
  • the desired product was prepared by substituting Fmoc-NMeAla for Fmoc- NMeNva in Example 1. Upon completion of the synthesis, cleavage of the peptide from the resin, removal of the protecting groups, and precipitation with diethyl ether, the crude peptide was obtained. This was purified by preparative HPLC using a C-l 8 column with a solvent system increasing in gradient from 5% to 100%) acetonitrile/water containing 0.01%) TFA over a period of 50 minutes.
  • the desired product was prepared by the procedure described in Example 1 with the following modification: coupling with acetic acid at the end of the synthesis was replaced with treatment of the peptide resin overnight with a 10-fold excess of succinic anhydride/pyridine in NMP. Upon completion of the synthesis, washing of the resin- bound peptide, cleavage of the peptide from the resin, removal of the protecting groups, and precipitation with diethyl ether, the crude peptide was obtained. This was purified by preparative HPLC using a C-l 8 column with a solvent system increasing in gradient from 5% to 100% acetonitrile/water containing 0.01% TFA over a period of 50 minutes.
  • Example 2 N-Ac-Sar-Glv-NMeAla-D-He-Thr-Nva-Ile-Arg-ProNH-ethyl
  • the procedure described in Example 1 was used substituting Fmoc-NMeAla for Fmoc-Val and Fmoc-Nva for Fmoc-NMeNva.
  • the crude peptide was obtained. This was purified by preparative HPLC using a C- 18 column and a solvent system increasing in gradient from 5% to 100% acetonitrile/water containing 0.01% TFA over a period of 50 minutes.
  • This can be purified by preparative HPLC using a C-l 8 column and a solvent system increasing in gradient from 5% to 100% acetonitrile/water containing 0.01%> TFA over a period of 50 minutes.
  • the pure fractions can be lyophilized to provide N-Ac-Sar-Gly-Val-D-alloIle-Thr-NMeVal-Ile- Arg-ProNH-ethyl as a trifluoroacetate salt.
  • NMeNva in Example 28.
  • the crude peptide was obtained. This was purified by preparative HPLC using a C-l 8 column and a solvent system increasing in gradient from 5%> to 100% acetonitrile/water containing 0.01% TFA over a period of 50 minutes.
  • EXAMPLE 30 N-Ac-Sar-Glv-Val-D-Ile-NMeThr-Nva-He-Arg-ProNH-ethyl
  • the desired product can be prepared by substituting Fmoc-NMeThr(t-Bu) for Fmoc-Thr(t-Bu) and Fmoc-Nva for Fmoc-NMeNva in Example 1.
  • Fmoc-NMeThr(t-Bu) for Fmoc-Thr(t-Bu)
  • Fmoc-Nva Fmoc-NMeNva
  • This can be purified purified by preparative HPLC using a C-l 8 column and a solvent system increasing in gradient from 5%> to 100% acetonitrile/water containing 0.01% TFA over a period of 50 minutes.
  • the pure fractions can then be lyophilized to provide N-Ac-Sar-Gly-Val-D-Ile- NMeThr-Nva-Ile-Arg-ProNH-ethyl as a trifluoroacetate salt.
  • the desired product can be prepared by substituting Fmoc-D-Leu for Fmoc-D-Ile, Fmoc-NMeSer(t-Bu) for Fmoc-Thr(t-Bu), and Fmoc-Nva for Fmoc-NMeNva in Example 1.
  • Fmoc-D-Leu for Fmoc-D-Ile
  • Fmoc-NMeSer(t-Bu) for Fmoc-Thr(t-Bu)
  • Fmoc-Nva for Fmoc-NMeNva
  • This can be purified by preparative HPLC using a C-l 8 column and a solvent system increasing in gradient from 5% to 100% acetonitrile/water containing 0.01% TFA over a period of 50 minutes.
  • the pure fractions can be lyophilized to provide N-Ac-Sar-Gly-Val- D-Leu-NMeSer-Nva-Ile-Arg-ProNH-ethyl as a trifluoroacetate salt.
  • This can be purified by preparative HPLC using a C-l 8 column and a solvent system increasing in gradient from 5% to 100% acetonitrile/water containing 0.01% TFA over a period of 50 minutes.
  • the pure fractions can be lyophilized to provide N-Ac-Sar-Gly-Val-D-Leu-Ser-NMeNva-Ile-Arg-ProNH- ethyl as a trifluoroacetate salt.
  • the desired product was prepared by substituting Fmoc-NMeSer(t-Bu) for Fmoc- NMeNva in Example 1. Upon completion of the synthesis, cleavage of the resin-bound peptide, removal of the protecting groups, and precipitation with diethyl ether, the crude peptide was obtained. This was purified by preparative HPLC using a C-l 8 column and a solvent system increasing in gradient from 5%> to 100% acetonitrile/water containing
  • This can be purified by preparative HPLC using a C-l 8 column and a solvent system increasing in gradient from 5% to 100% acetonitrile/water containing 0.01%> TFA over a period of 50 minutes.
  • the pure fractions can be lyophilized to provide N-Ac-Sar-Gly-Val-D-alloIle-Thr-NMeSer-Ile- Arg-Pro-D-AlaNH 2 as a trifluoroacetate salt.
  • EXAMPLE 38 N-Ac-Sar-Glv-Phe-D-Ile-Thr-NMeVal-Ile-Arg-Pro-D-AlaNH 2
  • the desired product can be prepared by substituting Fmoc-PheAla for Fmoc-Val and Fmoc-NMeVal for Fmoc-Nva in Example 28.
  • the crude peptide can be obtained.
  • This can be purified by preparative HPLC using a C-l 8 column and a solvent system increasing in gradient from 5% to 100% acetonitrile/water containing 0.01%> TFA over a period of 50 minutes.
  • the pure fractions can be lyophilized to provide N-Ac-Sar-Gly-Phe-D-Ile-Thr-NMeVal-Ile-Arg-Pro-D- AlaNH 2 as a trifluoroacetate salt.
  • the desired product can be prepared by substituting Fmoc-D-alloIle for Fmoc-DIle and Fmoc-Tyr(t-Bu) for Fmoc-Thr(t-Bu) in Example 1.
  • Fmoc-D-alloIle for Fmoc-DIle
  • Fmoc-Tyr(t-Bu) for Fmoc-Thr(t-Bu)
  • Example 1 Upon completion of the synthesis, cleavage of the resin-bound peptide, removal of the protecting groups, and precipitation with diethyl ether, the crude peptide can be obtained.
  • This can be purified by preparative HPLC using a C-l 8 column and a solvent system increasing in gradient from 5% to 100% acetonitrile/water containing 0.01% TFA over a period of 50 minutes.
  • the pure fractions can be lyophilized to provide N-Ac-Sar-Gly-Val-D-alloIle-Tyr-NMeNva-Ile-Arg-ProNH- ethyl as a trifluoroacetate salt.
  • the desired product can be prepared by substituting Fmoc-D-alloIle for Fmoc-D- Ile, Fmoc-Tyr(t-Bu) for Fmoc-Thr(t-Bu), and Fmoc-NMeVal for Fmoc-NMeNva in Example 1.
  • Fmoc-D-alloIle for Fmoc-D- Ile
  • Fmoc-Tyr(t-Bu) for Fmoc-Thr(t-Bu)
  • Fmoc-NMeVal for Fmoc-NMeNva
  • This can be purified by preparative HPLC using a C-l 8 column and a solvent system increasing in gradient from 5% to 100% acetonitrile/water containing 0.01% TFA over a period of 50 minutes.
  • the pure fractions can be lyophilized to provide N-Ac-Sar-Gly-Val-D-alloIle-Tyr-NMeVal-Ile-Arg-ProNH-ethyl as a trifluoroacetate salt.
  • the desired product can be prepared by substituting Fmoc-Gln(Trt) for Fmoc-Val in Example 28.
  • the crude peptide can be obtained. This can be purified by preparative HPLC using a C-l 8 column and a solvent system increasing in gradient from 5% to 100% acetonitrile/water containing 0.01 ) TFA over a period of 50 minutes.
  • the pure fractions can be lyophilized to provide N-Ac-Sar-Gly-Gln-D-Ile-Thr-NMeNva-Ile-Arg-Pro-D-AlaNH 2 as a trifluoroacetate salt.
  • This can be purified by preparative HPLC using a C-l 8 column and a solvent system increasing in gradient from 5% to 100% acetonitrile/water containing 0.01% TFA over a period of 50 minutes.
  • the pure fractions can be lyophilized to provide N-Ac-Sar-Gly-Val-D-alloIle-NMeThr-Nva-Ile-Arg-ProNH-ethyl as trifluoroacetate salt.
  • EXAMPLE 44 N-Ac-Sar-Glv-Val-D-Ile-Thr-NMeSer-He-Arg-Pro-D-AlaNH 2
  • the desired product can be prepared by Fmoc-NMeSer(t-Bu) for Fmoc-NMeNva in Example 28.
  • the crude peptide can be obtained.
  • This can be purified by preparative HPLC using a C-l 8 column and a solvent system increasing in gradient from 5% to 100%) acetonitrile/water containing 0.01% TFA over a period of 50 minutes.
  • the pure fractions can be lyophilized to provide N-Ac-Sar-Gly-Val-D-Ile-Thr-NMeSer-Ile-Arg-Pro-D-AlaNH 2 as trifluoroacetate salt.
  • the desired product can be prepared by substituting Fmoc-NMeVal for Fmoc-Val and Fmoc-Nva for Fmoc-NMeNva in Example 28.
  • the crude peptide can be obtained.
  • This can be purified by preparative HPLC using a C-l 8 column and a solvent system increasing in gradient from 5% to 100%> acetonitrile/water containing 0.01%) TFA over a period of 50 minutes.
  • the pure fractions can be lyophilized to provide N-Ac-Sar-Gly-NMeVal-D-Ile-Thr-Nva-Ile-Arg-Pro-D- AlaNH 2 as trifluoroacetate salt.
  • EXAMPLE 46 N-Ac-Sar-Glv-NMeVal-D-alloIle-Thr-Nva-He-Arg-ProNH-ethyl
  • the desired product can be prepared by substituting Fmoc-NMeVal for Fmoc-Val, Fmoc-D-alloIle for Fmoc-D-Ile, and Fmoc-Nva for Fmoc NMeNva in Example 1.
  • Fmoc-NMeVal for Fmoc-Val
  • Fmoc-D-alloIle for Fmoc-D-Ile
  • Fmoc-Nva Fmoc NMeNva
  • This can be purified by preparative HPLC using a C-l 8 column and a solvent system increasing in gradient from 5% to 100% acetonitrile/water containing 0.01% TFA over a period of 50 minutes.
  • the pure fractions can be lyophilized to provide N-Ac-Sar-Gly-NMeVal-D- alloIle-Thr-Nva-Ile-Arg-ProNH-ethyl as trifluoroacetate salt.
  • EXAMPLE 48 N-Ac-Sar-Glv-Val-D-HpheAla-Thr-NMeVal-He-Arg-ProNH-ethyl
  • the desired product can be prepared by substituting Fmoc-D-HpheAla for Fmoc- D-Ile and Fmoc-NMeVal for Fmoc-NMeNva in Example 1.
  • cleavage of the resin-bound peptide, removal of the protecting groups, and precipitation with diethyl ether the crude peptide can be obtained.
  • This can be purified by preparative HPLC using a C-l 8 column and a solvent system increasing in gradient from 5% to 100% acetonitrile/water containing 0.01% TFA over a period of 50 minutes.
  • the pure fractions can be lyophilized to provide N-Ac-Sar-Gly-Val-D-HpheAla-Thr-NMeVal- Ile-Arg-ProNH-ethyl as trifluoroacetate salt.
  • the desired product can be prepared by substituting Fmoc-D-Pen(SMe) for Fmoc- D-Ile and Fmoc-NMeVal for Fmoc-NMeNva in Example 1.
  • the crude peptide can be obtained.
  • This can be purified by preparative HPLC using a C-l 8 column and a solvent system increasing in gradient from 5%> to 100% acetonitrile/water containing 0.01% TFA over a period of 50 minutes.
  • the pure fractions can be lyophilized to provide N-Ac-Sar-Gly-Val-D-Pen(SMe)-Thr- NMeVal-Ile-Arg-ProNH-ethyl as trifluoroacetate salt.
  • the desired product can be prepared by substituting Fmoc-D-alloIle for Fmoc-D- Ile, Fmoc-NMeSer(t-Bu) for Fmoc-Thr(t-Bu), and Fmoc-Ser(t-Bu) for Fmoc-NMeNva in Example 1.
  • Fmoc-D-alloIle for Fmoc-D- Ile
  • Fmoc-NMeSer(t-Bu) for Fmoc-Thr(t-Bu)
  • Fmoc-Ser(t-Bu) for Fmoc-NMeNva
  • This can be purified by preparative HPLC using a C-l 8 column and a solvent system increasing in gradient from 5%> to 100% acetonitrile/water containing 0.01%) TFA over a period of 50 minutes.
  • the desired product was prepared by substituting Fmoc-Asn(Trt) for Fmoc-Val , Fmoc-D-Leu for Fmoc-D-Ile, Fmoc-NMeSer(t-Bu) for Fmoc-Thr(t-Bu), and Fmoc-Nva for Fmoc-NMeNva in Example 1.
  • Fmoc-Asn(Trt) for Fmoc-Val
  • Fmoc-D-Leu for Fmoc-D-Ile
  • Fmoc-NMeSer(t-Bu) for Fmoc-Thr(t-Bu)
  • Fmoc-Nva for Fmoc-NMeNva
  • the desired product can be prepared by substituting Fmoc-D-alloIle for Fmoc-D- Ile, Fmoc-NMeSer(t-Bu) for Fmoc-Thr(t-Bu), and Fmoc-Nva for Fmoc-NMeNva in Example 1.
  • Fmoc-D-alloIle for Fmoc-D- Ile
  • Fmoc-NMeSer(t-Bu) for Fmoc-Thr(t-Bu)
  • Fmoc-Nva for Fmoc-NMeNva
  • This can be purified by preparative HPLC using a C-l 8 column and a solvent system increasing in gradient from 5%> to 100% acetonitrile/water containing 0.01%) TFA over a period of 50 minutes.
  • the pure fractions can be lyophilized to provide NAc-Sar-Gly-Val-D-alloIle-NMeSer-Nva-Ile-Arg-ProNH-ethyl as trifluoroacetate salt.
  • the desired product was prepared by substituting Fmoc-Sar for Fmoc-Thr(t-Bu) and Fmoc-Nva for Fmoc-NMeNva in Example 1. Upon completion of the synthesis, cleavage of the resin-bound peptide, removal of the protecting groups, and precipitation with diethyl ether, the crude peptide was obtained. This was purified by preparative HPLC using a C-l 8 column and a solvent system increasing in gradient from 5% to 100%> acetonitrile/water containing 0.01%> TFA over a period of 50 minutes.
  • Fmoc-Sar for Fmoc-Thr(t-Bu)
  • Fmoc-Nva for Fmoc-NMeNva in Example 1.
  • the desired product was prepared by substituting Fmoc-NMe-D-Leu for NMeNva in Example 1. Upon completion of the synthesis, cleavage of the resin-bound peptide, removal of the protecting groups, and precipitation with diethyl ether, the crude peptide was obtained. This was purified by preparative HPLC using a C-l 8 column and a solvent system increasing in gradient from 5% to 100% acetonitrile/water containing 0.01% TFA over a period of 50 minutes.
  • the desired product was prepared by substituting Fmoc-Nva for Fmoc-NMeNva and Fmoc-NMeLeu for Fmoc-Ile in Example 1. Upon completion of the synthesis, cleavage of the resin-bound peptide, removal of the protecting groups, and precipitation with diethyl ether, the crude peptide was obtained. This was purified by preparative HPLC using a C-l 8 column and a solvent system increasing in gradient from 5% to 100% acetonitrile/water containing 0.01%> TFA over a period of 50 minutes.
  • the desired product was prepared by substituting Fmoc-D-alloIle for Fmoc-D-Ile, Fmoc-NMeSer(t-Bu) for Fmoc-Thr(t-Bu), and Fmoc-Ser(t-Bu) for Fmoc-NMeNva in Example 1.
  • Fmoc-D-alloIle for Fmoc-D-Ile
  • Fmoc-NMeSer(t-Bu) for Fmoc-Thr(t-Bu)
  • Fmoc-Ser(t-Bu) Fmoc-NMeNva
  • This can be purified by preparative HPLC using a C-l 8 column and a solvent system increasing in gradient from 5%> to 100% acetonitrile/water containing 0.01% TFA over a period of 50 minutes.
  • the pure fractions can be lyophilized to provide NAc-Sar-Gly-Val-D-alloIle-NMeTyr-Nva-Ile-Arg-ProHEt as trifluoroacetate salt.
  • EXAMPLE 100 NAc-Sar-Glv-Val-D-He-alloThr-NMeNva-Ile-Arg-ProNH-ethyl
  • the desired product can be prepared by substituting Fmoc-alloThr(t-Bu) for Fmoc- Thr(t-Bu) in Example 1.
  • the crude peptide can be obtained.
  • This can be purified by preparative HPLC using a C-l 8 column and a solvent system increasing in gradient from 5% to 100% acetonitrile/water containing 0.01%) TFA over a period of 50 minutes.
  • the pure fractions can be lyophilized to provide NAc-Sar-Gly-Val-D-Ile-alloThr-NMeNva-Ile-Arg-ProNH-ethyl as trifluoroacetate salt.
  • NAc-Sar-Glv-Gln-D-Ile-Thr-NMeNva-D-Ile-Arg-ProNH-ethyl The desired product can be prepared by substituting Fmoc-Gln(Trt) for Fmoc-Val and Fmoc-D-Ile for Fmoc-Ile in Example 1.
  • the crude peptide can be obtained.
  • This can be purified by preparative HPLC using a C-l 8 column and a solvent system increasing in gradient from 5%> to 100% acetonitrile/water containing 0.01% TFA over a period of 50 minutes.
  • the pure fractions can be lyophilized to provide NAc-Sar-Gly-Gln-D-Ile-Thr-NMeNva-D-Ile-Arg-ProNH- ethyl as trifluoroacetate salt.
  • EXAMPLE 102 NAc-Sar-Glv-Val-D-Ile-Thr-NMeNva-D-Lvs,Ac)-Arg-ProNH-ethyl
  • the desired product can be prepared by substituting Fmoc-D-Lys(Ac) for Fmoc-Ile in Example 1.
  • the crude peptide can be obtained.
  • This can be purified by preparative HPLC using a C-l 8 column and a solvent system increasing in gradient from 5% to 100% acetonitrile/water containing 0.01 ) TFA over a period of 50 minutes.
  • the pure fractions can be lyophilized to provide NAc-Sar-Gly-Val-D-Ile-Thr-NMeNva-D-Lys(Ac)-Arg-ProNH-ethyl as trifluoroacetate salt.
  • EXAMPLE 103 NAc-Sar-Glv-Gln-D-alloIle-NMeTyr-Nva-Ile-Arg-ProNH-ethyl
  • the desired product can be prepared by substituting Fmoc-Gln(Trt) for Fmoc-Val, Fmoc-D-alloIle for Fmoc-D-Ile, Fmoc-NMeTyr(t-Bu) for Fmoc-Thr(t-Bu), and Fmoc-Nva for Fmoc-NMeNva in Example 1.
  • the crude peptide can be obtained.
  • This can be purified by preparative HPLC using a C-l 8 column and a solvent system increasing in gradient from 5% to 100% acetonitrile/water containing 0.01% TFA over a period of 50 minutes.
  • the pure fractions can be lyophilized to provide NAc-Sar-Gly-Gln-D-alloIle-NMeTyr-Nva-Ile-Arg-ProNH-ethyl as trifluoroacetate salt.
  • the desired product can be prepared by substituting Fmoc-Gln(Trt) for Fmoc-Val, Fmoc-D-alloIle for Fmoc-D-Ile, Fmoc-NMeTyr(t-Bu) for Fmoc-Thr(t-Bu), Fmoc-Nva for Fmoc-NMeNva, and Fmoc-D-Ile for Fmoc-Ile in Example 1.
  • Fmoc-Gln(Trt) for Fmoc-Val
  • Fmoc-D-alloIle for Fmoc-D-Ile
  • Fmoc-NMeTyr(t-Bu) for Fmoc-Thr(t-Bu)
  • Fmoc-Nva for Fmoc-NMeNva
  • Fmoc-D-Ile for Fmoc-Ile in Example 1.
  • This can be purified by preparative HPLC using a C-l 8 column and a solvent system increasing in gradient from 5% to 100% acetonitrile/water containing 0.01% TFA over a period of 50 minutes.
  • the pure fractions can be lyophilized to provide NAc-Sar-Gly-Gln-D-alloIle-NMeTyr-Nva-D- Ile-Arg-ProNH-ethyl as trifluoroacetate salt.
  • NAc-Sar-Glv-Phe-D-He-Thr-NMeNva-Ile-Arg-Pro-D-AlaNH 2 The desired product can be prepared by substituting Fmoc-PheAla for Fmoc-Val in Example 28.
  • the crude peptide can be obtained.
  • This can be purified by preparative HPLC using a C-l 8 column and a solvent system increasing in gradient from 5% to 100% acetonitrile/water containing 0.01% TFA over a period of 50 minutes.
  • the pure fractions can be lyophilized to provide NAc-Sar-Gly-Phe-D-Ile-Thr-NMeNva-Ile-Arg-Pro-D-AlaNH 2 as trifluoroacetate salt.
  • NMePro-Glv-Ile-D-He-Thr-NMeNva-Ile-Arg-ProNH-ethyl The desired product can be prepared by substituting NMePro for Fmoc-Sar and Fmoc-Ile for Fmoc-Val in Example 1 , and omitting the final coupling with acetic acid. Upon completion of the synthesis, cleavage of the resin-bound peptide, removal of the protecting groups, and precipitation with diethyl ether, the crude peptide can be obtained.
  • This can be purified by preparative HPLC using a C-l 8 column and a solvent system increasing in gradient from 5%> to 100% acetonitrile/water containing 0.01% TFA over a period of 50 minutes.
  • the pure fractions can be lyophilized to NMePro-Gly-Ile-D-Ile-Thr- NMeNva-Ile-Arg-ProNH-ethyl as trifluoroacetate salt.

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PCT/US2000/032105 1999-11-22 2000-11-22 N-alkylated peptides having antiangiogenic activity WO2001038397A1 (en)

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CA002386893A CA2386893A1 (en) 1999-11-22 2000-11-22 N-alkylated peptides having antiangiogenic activity
JP2001540160A JP2003514920A (ja) 1999-11-22 2000-11-22 抗血管新生活性を有するｎ−アルキル化ペプチド
BR0010934-7A BR0010934A (pt) 1999-11-22 2000-11-22 Peptìdeos n-alquilados possuindo atividade antiangiogênica
EP00980685A EP1242455A1 (en) 1999-11-22 2000-11-22 N-alkylated peptides having antiangiogenic activity
AU17913/01A AU1791301A (en) 1999-11-22 2000-11-22 N-alkylated peptides having antiangiogenic activity
MXPA02005139A MXPA02005139A (es) 1999-11-22 2000-11-22 Peptidos n-alquilados que tienen actividad antiangiogenica.
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WO2003065974A2 (en) * 2001-10-31 2003-08-14 Abbott Laboratories Hexa-, hepta-, and octapeptides having antiangiogenic activity
EP1429796A2 (en) * 2001-04-11 2004-06-23 Abbott Laboratories Peptide antiangiogenic drugs
EP1531847A2 (en) * 2002-06-07 2005-05-25 Abbott Laboratories Pharmaceutical formulation
US7001984B2 (en) 2001-10-31 2006-02-21 Abbott Laboratories Di-, tri-, and tetra-peptides having antiangiogenic activity
US7037897B2 (en) 2001-10-31 2006-05-02 Abbott Laboratories TRI-, TETRA-, and penta-peptides having antiangiogenic activity
US7067490B2 (en) 2001-10-31 2006-06-27 Abbott Laboratories Hepta-, Octa-and nonapeptides having antiangiogenic activity
US7122625B2 (en) 2001-10-31 2006-10-17 Abbott Laboratories Hexa-, hepta-, and octapeptides having antiangiogenic activity
US7169888B2 (en) 2001-10-31 2007-01-30 Abbott Laboratories Tetra-, penta-, hexa- and heptapeptides having antiangiogenic activity
US7521425B2 (en) 2005-03-03 2009-04-21 Covx Technologies Ireland Limited Anti-angiogenic compounds
EP2177530A1 (en) 2001-10-31 2010-04-21 Abbott Laboratories Octapeptide having antiangiogenic activity

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WO1997033174A1 (en) * 1996-03-05 1997-09-12 Torrey Pines Institute For Molecular Studies Selectively n-alkylated peptidomimetic combinatorial libraries and compounds therein
WO1997041824A2 (en) * 1996-05-03 1997-11-13 Abbott Laboratories Antiangiogenic peptides derived from plasminogen
WO1998041542A1 (en) * 1997-03-17 1998-09-24 Abbott Laboratories Antiangiogenic drug to treat cancer, arthritis and retinopathy

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1997033174A1 (en) * 1996-03-05 1997-09-12 Torrey Pines Institute For Molecular Studies Selectively n-alkylated peptidomimetic combinatorial libraries and compounds therein
WO1997041824A2 (en) * 1996-05-03 1997-11-13 Abbott Laboratories Antiangiogenic peptides derived from plasminogen
WO1998041542A1 (en) * 1997-03-17 1998-09-24 Abbott Laboratories Antiangiogenic drug to treat cancer, arthritis and retinopathy

Cited By (15)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP1429796A4 (en) * 2001-04-11 2005-01-12 Abbott Lab ANTIANGIOGENIC DRUGS BASED ON PEPTIDES
EP1429796A2 (en) * 2001-04-11 2004-06-23 Abbott Laboratories Peptide antiangiogenic drugs
US7001984B2 (en) 2001-10-31 2006-02-21 Abbott Laboratories Di-, tri-, and tetra-peptides having antiangiogenic activity
WO2003065974A3 (en) * 2001-10-31 2004-07-15 Abbott Lab Hexa-, hepta-, and octapeptides having antiangiogenic activity
WO2003065974A2 (en) * 2001-10-31 2003-08-14 Abbott Laboratories Hexa-, hepta-, and octapeptides having antiangiogenic activity
US7037897B2 (en) 2001-10-31 2006-05-02 Abbott Laboratories TRI-, TETRA-, and penta-peptides having antiangiogenic activity
US7067490B2 (en) 2001-10-31 2006-06-27 Abbott Laboratories Hepta-, Octa-and nonapeptides having antiangiogenic activity
US7122625B2 (en) 2001-10-31 2006-10-17 Abbott Laboratories Hexa-, hepta-, and octapeptides having antiangiogenic activity
US7169888B2 (en) 2001-10-31 2007-01-30 Abbott Laboratories Tetra-, penta-, hexa- and heptapeptides having antiangiogenic activity
EP2177530A1 (en) 2001-10-31 2010-04-21 Abbott Laboratories Octapeptide having antiangiogenic activity
EP1531847A2 (en) * 2002-06-07 2005-05-25 Abbott Laboratories Pharmaceutical formulation
JP2005538960A (ja) * 2002-06-07 2005-12-22 アボット・ラボラトリーズ 医薬製剤
EP1531847A4 (en) * 2002-06-07 2009-07-01 Abbott Lab PHARMACEUTICAL FORMULATION
US7521425B2 (en) 2005-03-03 2009-04-21 Covx Technologies Ireland Limited Anti-angiogenic compounds
EP2292248A2 (en) 2005-03-03 2011-03-09 CovX Technologies Ireland Limited Anti-angiogenic compounds

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