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WO1995020603A1 - Inhibiteurs de serine proteases portant un groupe chelateur - Google Patents

Inhibiteurs de serine proteases portant un groupe chelateur Download PDF

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Publication number
WO1995020603A1
WO1995020603A1 PCT/EP1995/000269 EP9500269W WO9520603A1 WO 1995020603 A1 WO1995020603 A1 WO 1995020603A1 EP 9500269 W EP9500269 W EP 9500269W WO 9520603 A1 WO9520603 A1 WO 9520603A1
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Prior art keywords
peptide
group
compound
chelating
alkyl
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PCT/EP1995/000269
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English (en)
Inventor
Rainer Metternich
Carlo Tapparelli
Anette Wienand
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Sandoz Ltd.
Sandoz-Patent-Gmbh
Sandoz-Erfindungen Verwaltungsgesellschaft Mbh
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Sandoz Ltd., Sandoz-Patent-Gmbh, Sandoz-Erfindungen Verwaltungsgesellschaft Mbh filed Critical Sandoz Ltd.
Priority to JP7519884A priority Critical patent/JPH09509653A/ja
Priority to EP95907611A priority patent/EP0741747A1/fr
Priority to AU15764/95A priority patent/AU1576495A/en
Publication of WO1995020603A1 publication Critical patent/WO1995020603A1/fr

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K51/00—Preparations containing radioactive substances for use in therapy or testing in vivo
    • A61K51/02—Preparations containing radioactive substances for use in therapy or testing in vivo characterised by the carrier, i.e. characterised by the agent or material covalently linked or complexing the radioactive nucleus
    • A61K51/04—Organic compounds
    • A61K51/08—Peptides, e.g. proteins, carriers being peptides, polyamino acids, proteins
    • A61K51/088—Peptides, e.g. proteins, carriers being peptides, polyamino acids, proteins conjugates with carriers being peptides, polyamino acids or proteins
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
    • C07K5/06—Dipeptides
    • C07K5/06191—Dipeptides containing heteroatoms different from O, S, or N
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
    • C07K5/08—Tripeptides
    • C07K5/0827—Tripeptides containing heteroatoms different from O, S, or N
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides

Definitions

  • This invention relates to inhibitors of serine proteases, e.g. thrombin, and conjugates of such inhibitors used as thrombus imaging agents.
  • thrombin serine proteases
  • conjugates of such inhibitors used as thrombus imaging agents Thrombin and other serine proteases and inhibitors therefor are discussed in WO 94/20526, the disclosure of which is incorporated herein by reference.
  • transition state analogues are meant compounds as defined by A.J. Barrett et al. in Proteinase Inhibitors, Ed. Elsevier Amsterdam, 1986, p. 57.
  • pseudo-peptide or "pseudo-peptidic” as used herein refers to a peptide isostere in which the peptide bond between 2 amino acid residues, whether natural or unnatur ⁇ l, is replaced with any isosteric group, e.g. -CH 2 -NH-.
  • the pseudo-peptide may comprise one or more such isosteric bonds.
  • the chelating group is linked by a covalent bond to the amino group of the peptide or pseudo-peptide.
  • the chelating group is preferably attached to the N-terminal amino group of the peptide or pseudo-peptide.
  • the chelating group may be attached either directly or indirectly, e.g. by means of a spacer group, to the amino group of the peptide or pseudo-peptide.
  • the chelating group is coupled to the peptide or pseudo-peptide or to the peptide-spacer or (pseudo-peptide)-spacer moiety by an amide bond.
  • Preferred compounds of the invention are those comprising a C-terminal boronic acid derivative residue, particularly a C-terminal boronic acid derivative of lysine, ornithine, arginine or an aliphatic amino alcohol. More preferred are those comprising additionally at least one unnatural ⁇ -amino acid having a hydrophobic side chain.
  • the present invention provides a compound of formula I wherein
  • Y 1 and Y 2 independently, are OH or F or, taken together, form a moiety derived from a dihydroxy compound having at least two hydroxy groups separated by at least two connecting atoms in a chain or ring, said chain or ring comprising 1 to about 20 carbon atoms and, optionally, a heteroatom which can be N, S, or O;
  • R 1 is a substituted alkyl selected from the group consisting of -(CH 2 ) z -X, -CH(CH 3 )-(CH 2 ) 2 -X, -CH 2 -CH(CH 3 )-CH 2 -X, -(CH 2 ) 2 -CH(CH 3 )-X, and -(CH 2 ) 2 -C(CH 3 ) 2 -X, where X is -NH 2 , -NH-C(NH)-NH 2 or -S-C(NH)-NH 2 , and z is 3 to 5; m is 1 or 2; r, o, p and q are, independently, either 1 or 0; A 1 , A 2 and A 3 , independently, are amino acids of L- or D-configuration selected from the group consisting of Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro,
  • W is a chelating group attached to the N-terminal amino group of the compound
  • Z is a direct bond or a spacer in free form or in salt form.
  • a particularly preferred compound of formula I is
  • Y is a sequence of n amino acids selected such that the n+1 amino acid peptide Y-Lys or Y-Arg has an affinity for the active site of a trypsin-like protease, where n is an integer from 1 to 10 preferably 1 to 4, and in which at least one amino acid is an unnatural amino acid having a hydrophobic side chain;
  • Q 1 and Q 2 which may be the same or different, are selected from -OH, COR 2 , -CONR 2 R 3 , -NR 2 R 3 , and -OR 6 , or Q 1 and Q 2 taken together form a diol residue, and wherein R 2 , R 3 and R 6 , which may be the same or different, are Ci.ioalkyl, C 6-10 aryl, C 6-10 aralkyl, or phenyl substituted by up to three groups selected from C 1-4 alkyl, halogen and C 1-4 alkoxy; R 4 is hydrogen or C 1-10 alkyl ;
  • R 5 is a group -Z 1 -X 1 ,
  • Z 1 is -(CH 2 ) s -; -CH(CH 3 )-(CH 2 ) 2 - , -CH 2 -CH(CH 3 )-CH 2 -, -(CH 2 ) 2 -CH(CH 3 )-, -(CH 2 ) 2 -C(CH 3 ) 2 - , -CH(CH 3 ) - (CH 2 ) 3 -, -CH 2 -CH(CH 3 ) - (CH 2 ) 2 -, -(CH 2 ) 2 -CH(CH 3 )-CH 2 -, -(CH 2 ) 3 -C(CH 3 ) 2 -, - (CH 2 ) 2 -C(CH 3 ) 2 -, - (CH 2 ) 2 -C(CH 3 ) 2 -, - (CH 2 ) 2 -C(CH 3 ) 2 -, - (CH 2 ) 2 -C(CH 3 ) 2 -
  • s 2, 3, 4 or 5;
  • X 1 is -NH 2 , -NH-C(NH)-NH 2 , -S-C(NH)-NH 2 , -N 3 , C 1-4 alkoxy, C 1-4 alkylthio, -Si(CH 3 ) 3 , OH, SH, NR 7 R 8 , or phenyl, optionally substituted by up to three groups selected from halogen, OH and C 1-4 alkoxy, or C(NH)R 10 ;
  • R 7 is H or C 1-10 alkyl
  • R 8 is C 1-10 alkyl, -CO-
  • R 9 or CS-R 9 R 10 is C 1-3 alkyl or N(CH 3 ) 2 , or NR 11 R 12 ; wherein R 9 is H or C 1-10 alkyl, C 1-10 alkoxy, C 6- 10 aryl, C 6-10 aralkyl and R 11 and R 12 (which may be the same or different) are H or C 1-10 alkyl; or R 4 and R 5 together form a trimethylene group, and the asymmetric carbon atom marked * may have the D- or L-configuration, or represent any mixture of these.
  • an unnatural amino acid is meant any amino acid other than D- or L- Ala, Arg, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val.
  • Preferred unnatural amino acids are of formula III
  • R 18 is a hydrophobic group.
  • Preferred hydrophobic groups consist of a methylene group linked to an aromatic group optionally substituted by a polar group, to an alicyclic group having at least two rings and no polar substituents, or to a tert.butyl or trimethylsilyl group.
  • R 18 is R 18 ' where R 18 ' is a group of formulae c), d), e), f), g), h) or i)
  • the unnatural amino acids of formula III may be in D- or L-form or any mixture of these, but are preferably in D-form.
  • Particularly preferred compounds of formula II are those compounds in which Q 1 and Q 2 taken together represent the group OPin of formula a).
  • the amino acids constituting Y are ⁇ -amino acids which may be selected from the L-amino acids naturally occurring in proteins, their corresponding enantiomeric D-amino acids or chemically modified alpha-amino acids such as glutamic acid gamma-piperidide:
  • pipecolic acid provided that at least one amino acid is an unnatural amino acid having a hydrophobic side chain.
  • More preferred compounds of formula II are those in which Y is a sequence of two amino acids, of which the N-terminal amino acid is the unnatural amino acid and the other amino acid is L-proline (Pro), e.g. a compound of formula IIa
  • X 1 ' is -NH 2 , -NH-C(NH)-NH 2 , -N 3 , OH -Si(CH 3 ) 3 , phenyl, monochloro-phenyl, dimethylamino-phenyl, -NH- C(O)-CH 3 , -NHC(O)H, -NHC(S)-NHCH 3 , -NH-C(O)-CH(CH 3 ) 2 ,
  • Suitable chelating groups e.g. as W are physiologically acceptable chelating groups capable of complexing a detectable element.
  • the chelating group has a substantially hydrophilic character.
  • chelating groups include iminodicarboxylic groups or polyaminopolycarboxylic groups, e.g. those derived from non cyclic ligands such as ethylene diaminetetraacetic acid (EDTA), diethylene triamine pentaacetic acid (DTPA), ethylene glycol-O,O'-bis(2-aminoethyl)-N,N,N',N'-tetraacetic acid (EGTA), N,N'-bis(hydroxybenzyl)ethylenediamine-N,N'-diacetic acid (HBED) and triethylenetetramine hexaacetic acid (TTHA); those derived from substituted-EDTA or -DTPA, such as p-isothiocyanato-benzyl-EDTA or -DTPA, p-amino-benzyl-DTPA or ⁇ -amino-C 1-4 alkylene-DTPA;
  • 1,4,7,10-tetra azacyclo-dodecane-N,N',N",N'''-tetraacetic acid DAA
  • 1,4,7,10-tetra-azacyclotridecane-1,4,7,10-tetraacetic acid TITRA
  • 1,4,8,11-tetraazacyclotetradecane-N,N',N",N'''-tetraacetic acid TETA
  • Chelating groups particularly suitable for Tc labelling may be any of the known chelating groups capable of complexing 99m Tc, e.g. as disclosed in EP-A-279, 417, DOS 41 28 183 or WO 92/05154, the disclosures of which are incorporated herein by reference. These complexing groups may be linked to the peptide or pseudo-peptide via a group of formula - (R 20 ) n -R 21 wherein n 1 is 0 or 1, R 20 is C 1-6 -alkylene, -O-C 1-6 alkylene or -NH-C 1-6 alkylene and R 21 is -NCS, a carboxy group or a functional derivative thereof, e.g. acid halide, anhydride or hydrazide, optionally through a spacer group.
  • Preferred chelating groups are radicals of formula (j)
  • W' is a chelating group derived from a non-cyclic or cyclic polyamino-polyacetic acid or anhydride
  • X a is phenylene or C 1-3 alkylene.
  • chelating groups are preferred as chelating groups.
  • Suitable spacer groups include groups of formula (k)
  • R 22 is a divalent residue derived from a functional moiety capable of covalently reacting with the chelating agent
  • R 23 is C 1-11 alkylene optionally interrupted by one or more heteroatoms or residues selected from O, S, CO, -NH-, -NHCO-, N(C 1-4 alkyl)-CO- and -N(C 1-4 alkyl)-, C 2-11 alkenylene or -CH(R 24 )- wherein R 24 is a residue attached to a natural or unnatural ⁇ -amino acid, e.g. hydrogen, C 1-11 alkyl, benzyl, optionally substituted benzyl, naphthyl-methyl, pyridyl-methyl.
  • m is preferably 1.
  • the compounds of the invention may exist in free or salt form.
  • Salts include acid addition salts, e.g. with organic acids, polymeric acids or inorganic acids; for example, hydrochlorides and acetates, and salt forms obtainable with the carboxylic acid groups present in the chelating group, e.g. alkali metal salts such as sodium or potassium, or substituted or unsubstituted ammonium salts.
  • the present invention also includes a process for the production of the compounds of the invention. They may be produced by analogy to known methods.
  • the compounds of the invention may be produced for example as follows: a) removing at least one protecting group which is present in a trypsin-like serine protease inhibiting peptide of the invention bearing a protecting group; b) linking together by an amide bond two peptide fragments each of them containing at least one amino acid in protected or unprotected form and one of them containing the chelating group, wherein the amide bond is such that the desired amino acid sequence is obtained, and then optionally effecting step a) above; c) linking together a chelating agent and the desired peptide in protected or unprotected form in such a way that the chelating group is attached to the desired amino group of the peptide, and then optionally effecting step a) above; or d) removing or converting a functional group of an unprotected or a protected peptide bearing a chelating
  • the above reactions may be effected analogously to known methods, e.g. as described in the following examples, in particular process steps a) and c) above.
  • the chelating group is attached by an amide bond, this may be carried out analogously to the methods used for amide formation.
  • protecting groups which are suitable for use in peptides or for the desired chelating groups may be used for functional groups which do not participate in the reaction.
  • the term protecting group may also include a polymer resin having functional groups.
  • the peptide fragment bearing the chelating group and used in step b) above may be prepared by reacting the peptide fragment comprising at least one amino acid in protected or unprotected form with the chelating agent.
  • the reaction may be performed by analogy with step c).
  • the chelating agent used in process step c) may be known or prepared analogously to known procedures.
  • the compound used is such that it allows the introduction of the desired chelating group on the peptide, e.g. a polyaminopolycarboxylic acid as disclosed, a salt or anhydride thereof.
  • amino-acids, peptide fragments or peptides used as starting materials which have the chelating group attached through a spacer group to the peptide, e.g. a radical of formula (j) as defined above, may be prepared by reacting the corresponding amino-acids or peptides free from spacer group in conventional manner with a corresponding spacer-yielding compound, such as an acid or reactive acid derivative comprising the spacer group.
  • a corresponding spacer-yielding compound such as an acid or reactive acid derivative comprising the spacer group.
  • an acid of formula R' 22 -R 23 -COOH or a reactive acid derivative thereof such as an active ester, wherein R' 22 is a functional moiety capable of covalently reacting with the chelating agent may be used.
  • active ester groups or carboxy activating groups are e.g. 4-nitrophenyl, pentachlorophenyl, pentafluorophenyl, succinimidyl or 1-hydroxy-benz
  • the chelating agent may be reacted first with a spacer group-yielding compound, in order to bear the spacer group and then be reacted in conventional manner with the peptide, peptide fragment or amino-acid.
  • the peptides or peptide fragments used as starting materials in process steps b), c) and d) may be prepared e.g. as disclosed in EP-A2-293, 881, EP-A2-471,561, or WO 94/20526.
  • the compounds of the invention may be purified in conventional manner, e.g. by chromatography.
  • the compounds of the invention contain less than 5% by weight of peptides free of chelating groups.
  • the compounds of the invention exhibit pharmacological activity and are therefore pharmaceutically useful.
  • the compounds of the invention have trypsin-like serine protease inhibiting properties, as indicated in the test methods descibed in WO 94/20526
  • the compounds of the invention are useful as inhibitors of trypsin-like serine proteases and may be employed and administered as described in WO 94/20526.
  • the compounds of the invention may be administered in free form or in pharmaceutically acceptable salt form. Such salts may be prepared in conventional manner and exhibit the same order of activity as the free compounds.
  • the present invention further provides: a) a compound of the invention or a pharmaceutically acceptable salt thereof for use as a pharmaceutical; b) a method of preventing or treating disorders caused by an excess of one or more trypsin-like serine protease in a subject in need of such a treatment, which method comprises administering to said subject an effective amount of a compound of the invention or a pharmaceutically acceptable salt thereof; c) a pharmaceutical composition comprising a compound of the invention in free form or in pharmaceutically acceptable salt form in association with a pharmaceutically acceptable diluent or carrier.
  • the compounds of the invention can be complexed with a detectable element.
  • the present invention also provides the compounds of the invention as defined above which are complexed with a detectable element (hereinafter referred to as chelates of the invention), in free form or in salt form, their preparation and their use as diagnostic agents.
  • a detectable element hereinafter referred to as chelates of the invention
  • detectable element any element, preferably a metal ion which exhibits a property detectable in diagnostic techniques, e.g. a metal ion which emits a detectable radiation or a metal ion which is capable of influencing NMR relaxation properties.
  • Suitable detectable metal ions include for example paramagnetic ions, e.g. Gd 3+ , Fe 3+ , Mn 2+ and Cr 2+ , fluorescent metal ions, e.g. Eu 3+ , and radionuclides, e.g. ⁇ -emitting radionuclides, positron-emitting radionuclides e.g. 68 Ga.
  • Suitable T-emitting radionuclides include those which are useful in diagnostic techniques.
  • the T-emitting radionuclides advantageously have a half-life of from 1 hour to 40 days, preferably from 5 hours to 4 days, more preferably from 5 hours to 3 days and preferably should exhibit a single T-emission, preferably in the range of 100-200 KeV.
  • Examples are radionuclides derived from Gallium, Indium, Technetium, Ytterbium, Rhenium and Thallium e.g. 67 Ga, 111 In, 99m Tc, 169 Yb.
  • the chelates of the invention may be prepared by reacting the compound with a corresponding detectable element yielding compound, e.g. a metal salt, preferably a water-soluble salt.
  • a metal salt e.g. a water-soluble salt.
  • the reaction may be carried out by analogy with known methods, e.g. as disclosed in Perrin, Organic Ligand, Chemical Data Series 22. NY Pergamon Press (1982); in Krejcarit and Tucker, Biophys. Biochem. Res. Com. 77: 581 (1977) and in Wagner and Welch, J. Nucl. Med. 20: 428 (1979).
  • the complexing of a compound of the invention is effected at a pH at which it is stable.
  • the detectable element may also be provided to the solution as a complex with an intermediate chelating agent, e.g. a chelating agent which forms a chelate complex that renders the element soluble at the physiological pH of the complex but is less thermodynamically stable than the chelate.
  • an intermediate chelating agent e.g. a chelating agent which forms a chelate complex that renders the element soluble at the physiological pH of the complex but is less thermodynamically stable than the chelate.
  • an intermediate chelating agent is 4,5-dihydroxy-1,3-benzene-disulfonic acid (Tiron).
  • Tiron 4,5-dihydroxy-1,3-benzene-disulfonic acid
  • the above mentioned reactions may be effected analogously to known methods.
  • the above mentioned reactions are conveniently effected under conditions avoiding trace metal contamination.
  • distilled de-ionized water, ultrapure reagents, no carrier-added radioactivity etc. are used to reduce the effects of trace metal.
  • the chelates of the invention may exist e.g. in free or salt form.
  • Salts include acid addition salts with e.g. organic acids, polymeric acids or inorganic acids, for example hydrochlorides and acetates, and salt forms obtainable with the carboxylic acid groups present in the molecule which do not participate to the chelate formation.
  • the chelates of the invention and their pharmaceutical acceptable salts exhibit diagnostic activity and are therefore useful as imaging agents, e.g. visualisation of thrombi. They exhibit significant serum stability and binding affinity to thrombin, as indicated in standard in vitro tests.
  • the compound of Example 4 has a half life of 210 hours in human serum, when tested in vitro.
  • the present invention also provides a method for in vivo detection of a thrombus in a subject which comprises a) administering a chelate of the invention to said subject and b) recording the localisation of the thrombus targeted by said chelate.
  • the chelates of the invention for use as an imaging agent in the above method may be administered parenterally, preferably intravenously, e.g. in the form of injectable solutions or suspensions, preferably in a single injection.
  • the appropriate dosage will of course vary depending upon, for example, the compound and the type of detectable element used.
  • a suitable dose to be injected is in the range to enable imaging by photoscanning procedures known in the art.
  • a chelate of the invention may advantageously be administered in a dose comprising 0.1 to 10 mCi of a stably chelated radionuclide.
  • an indicated dosage range may be of from 0.1 to 1 ⁇ g/kg of a compound of the invention labeled with 0.02 to 0.5 mCi T-emitting radionuclide.
  • an indicated dosage range may be of from 1 to 20 ⁇ g compound labeled e.g. with 3 to 10 mCi T-emitting radionuclide, preferably 2 to 5 mCi.
  • the localisation of a thrombus with the chelates may be followed by the corresponding imaging techniques, e.g.
  • nuclear medicine imaging instrumentation for example a scanner, a conventional gamma camera to obtain planar images, a rotating T-camera to perform Single Photon Emission Computer Tomography (SPECT) or suitable instrumentation to perform Photon Emission Tomography (PET), each preferably computer assisted.
  • SPECT Single Photon Emission Computer Tomography
  • PET Photon Emission Tomography
  • the chelates of the invention may be administered in free form or in pharmaceutically acceptable salt form. Such salts may be prepared in conventional manner and exhibit the same order of activity as the free compounds.
  • the chelates of the invention are particularly useful in detecting thrombi formation resulting from vascular thrombotic disorders caused by myocardial infarction, artheriosclerosis, stroke, disseminated intravascular coagulation, peri- and post-operative to surgery, vascular recanalisations, immobility, atrial fibrillation and heart insufficiency.
  • the chelates of the invention for use in the method of the present invention may preferably be prepared shortly before administration to a subject, i.e. the radiolabeling with the desired detectable metal ion, may be performed shortly before administration.
  • a pharmaceutical composition comprising a chelate according to the invention in free or in pharmaceutically acceptable salt form, together with one or more pharmaceutically acceptable carriers or diluents therefor.
  • Such a composition may be manufactured in conventional manner.
  • a composition according to the invention may also be presented in separate package with instructions for mixing the compound with the metal ion and for the administration of the resulting chelate. It may also be presented in twin-pack form, that is, as a single package containing separate unit dosages of the compound and the detectable metal ion with instructions for mixing them and for administration of the chelate. A diluent or carrier may be present in the unit dosage forms.
  • Boc t-butyloxycarbonyl
  • Baa -NH-CH- (CH 2 CH 2 CH 2 Br)B-
  • H-D-TMSal-Pro-Boro (e -N-Fmoc) Lys-OPin (trifluoroacetate) (428.4 mg, 0.5 mMol) is dissolved in dioxane and is treated with 5.0 ml saturated aqueous NaHCO 3 and DTPA-anhydride (0.268 g, 0.75 mMol). The mixture is stirred for 0.5 h at room temperature. Acetic acid (35.0 ml) is added and the mixture is concentrated in vacuo at 40°. The resulting residue is treated with 50 ml EtOAc and H 2 O. The aqueous layer is extracted with EtOAc and the combined organic layers are washed with H 2 O.
  • DTPA-(D-TMSal-Pro-Boro(e-N-Fmoc)Lys-OPin) 2 (1.92 g, 1.042 mMol) of step a) is treated with 5.0 ml piperidine in DMF (20 % solution) at room temperature for 15 min. The resulting suspension is diluted with 50 ml of ether and the desired product precipitates as a crude material. This material is purified over Dowex (1-X4, 200-400 mesh), EtOH/H 2 O (1:1) and 10 % AcOH. After removal of the solvent the desired product is obtained as a white foam and is crystallized from EtOH/ether to give the title compound as an amorphous white solid.
  • H-D-TMSal-Pro-Boro( ⁇ -N-Fmoc) Lys-OPin(trifluoroacetate), used as starting material, may be prepared as follows:
  • Boc-D-TMSal-Pro-BoroLys-OPin(620.7 mg,1.0 mMol) and Fmoc-OSu (337.3 mg, 1.0 mMol) in dioxane are treated at room temperature with saturated aqueous NaHCO 3 solution. After 45 min, the reaction mixture is diluted with EtOAc and is washed with water and brine. After drying over Na 2 SO 4 and concentration in vacuo, the title compound is used without further purification in the next step.
  • step B) The product of step B) (2.36 g, 2.8 mMol) is treated with 8.5 ml TFA/H 2 O (95:5) and 0.85 ml anisol. After 1 h at room temperature, the reaction mixture is concentrated in vacuo and the resulting oil is chromatographed on a 3.2 ⁇ 25 cm HPLC column RP18 (10 ⁇ m), H 2 O/MeOH/TFA. The desired product is obtained as a white foam, MS: 743 (M + H) + EXAMPLE 2: 111 In labeled compound of Example 1
  • Example 1 1 mg of compound of Example 1 is dissolved in 5 ml 0.01 M acetic acid. The resulting solution is passed through a 0.22 ⁇ Millex-GV filter and dispensed in 0.1 ml portions and stored at -20°C. 111 InCl 3 (Amersham, 1 mCi/100 ⁇ l) is prediluted in an equal volume of 0.5 M sodium acetate and labeling is carried out by mixing the ligand with the InCl 3 solution and gentle homogenisation at room temperature. HEPES buffer, pH 7.4, is then added to make a 10 -6 M solution.
  • HEPES buffer pH 7.4
  • step C HOOC(CH 2 ) 2 CO-D-TMSal-Pro-Boro( ⁇ -N-Fmoc)Lys-OPin H-D-TMSal-Pro-Boro( ⁇ -N-Fmoc)Lys-OPin (trifluoro-acetate of Example 1, step C) (0.857 g,1.0 mMol) is dissolved in dioxane and is treated at room temperature with saturated aqueous NaHCO 3 solution and succinic anhydride (0.11 g, 1.10 mMol). After 45 min, the reaction mixture is diluted with EtOAc and is washed with aqueous NaHSO 4 -solution (1 M) and brine.
  • step a) The product of step a) (623 mg, 0.739 mMol) in THF is cooled to 0° and is treated with N-methyl morpholine (0.081 ml, 0.739 mMol and pivaloyl chloride (0.095 ml, 0.776 mMol) to form the mixed anhydride. After stirring for 1 h at 0°, the reaction mixture is treated with ( t Bu)-DTPA-(p-NH 2 -Bzl) (575 mg, 0.739 mMol) (as described in Bioconjugate Chem. 1991, 2(3), 180-186) and additional N-methyl morpholine (0.081 ml, 0.739 mMol).
  • step b) The product of step b) (576 mg, 0.296 mMol) is saponified by treatment with TFA/H 2 O (95:5) in the presence of anisol/ethanedithiol (1:1) for 7 h at room temperature. Excess TFA is removed in vacuo and the resulting residue is treated with ether to precipitate the desired product. After filtration and drying under reduced pressure, the title compound is obtained as an amorphous solid foam of the trifluoroacetate MS: 1323 (M + H) + . d DTPA-(p-NH-Bzl)-CO(CH 2 ) 2 -CO-D-TMSal-Pro-BoroLys- OPin
  • the 111 In labeled compound is prepared by using the same procedure as disclosed in Example 2.

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  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)

Abstract

L'invention se rapporte à un peptide inhibant une sérine protéase analogue à la trypsine ou un pseudo-peptide portant un groupe chélateur capable de complexer un élément détectable, le groupe chélateur étant lié à un groupe amino du peptide ou du pseudo-peptide de sorte que ce groupe chélateur n'interfère pas avec la thrombine ou empêche la liaison du peptide ou du pseudo-peptide à celle-ci. L'invention se rapporte de préférence à un composé de la formule (I) ou (II) se présentant sous une forme libre, sous forme de sel ou sous forme d'un complexe, par ex. un complexe avec un élément détectable, à la préparation de ce composé, à son utilisation comme médicament ou, lorsqu'il est complexé avec un radionucléide émettant des rayons Ω, comme produit radiopharmaceutique, par ex. pour être utilisé dans la visualisation de thrombus, et à des compositions pharmaceutiques contenant ce composé.
PCT/EP1995/000269 1994-01-26 1995-01-25 Inhibiteurs de serine proteases portant un groupe chelateur WO1995020603A1 (fr)

Priority Applications (3)

Application Number Priority Date Filing Date Title
JP7519884A JPH09509653A (ja) 1994-01-26 1995-01-25 キレート基を有するセリンプロテアーゼ阻害剤
EP95907611A EP0741747A1 (fr) 1994-01-26 1995-01-25 Inhibiteurs de serine proteases portant un groupe chelateur
AU15764/95A AU1576495A (en) 1994-01-26 1995-01-25 Inhibitors of serine proteases, bearing a chelating group

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
GB9401483A GB9401483D0 (en) 1994-01-26 1994-01-26 Organic compounds
GB9401483.4 1994-01-26

Publications (1)

Publication Number Publication Date
WO1995020603A1 true WO1995020603A1 (fr) 1995-08-03

Family

ID=10749357

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/EP1995/000269 WO1995020603A1 (fr) 1994-01-26 1995-01-25 Inhibiteurs de serine proteases portant un groupe chelateur

Country Status (10)

Country Link
EP (1) EP0741747A1 (fr)
JP (1) JPH09509653A (fr)
AU (1) AU1576495A (fr)
CA (1) CA2179889A1 (fr)
GB (1) GB9401483D0 (fr)
IL (1) IL112423A0 (fr)
PE (1) PE45295A1 (fr)
SG (1) SG47079A1 (fr)
WO (1) WO1995020603A1 (fr)
ZA (1) ZA95636B (fr)

Cited By (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1997005161A1 (fr) * 1995-07-28 1997-02-13 Novartis Ag Derives d'acides bores
WO2002022011A1 (fr) 2000-09-15 2002-03-21 Institut für Diagnostikforschung GmbH an der Freien Universität Berlin Procede de representation par images et de diagnostic de thrombus par tomographie a spin nucleaire faisant appel a des agents de contraste particulaires
EP1396269A1 (fr) 2002-09-09 2004-03-10 Trigen Limited Sels d'acide boronique des metaux multivalents et leur utilisation dans la préparation de médicaments pour le traitement de la thrombose
US7223745B2 (en) 2003-08-14 2007-05-29 Cephalon, Inc. Proteasome inhibitors and methods of using the same
US7576206B2 (en) 2003-08-14 2009-08-18 Cephalon, Inc. Proteasome inhibitors and methods of using the same
US8283367B2 (en) 2005-02-11 2012-10-09 Cephalon, Inc. Proteasome inhibitors and methods of using the same
US8541590B2 (en) 2009-12-22 2013-09-24 Cephalon, Inc. Proteasome inhibitors and processes for their preparation, purification and use
US11267803B2 (en) 2016-06-21 2022-03-08 Orion Ophthalmology LLC Carbocyclic prolinamide derivatives
US11377439B2 (en) 2016-06-21 2022-07-05 Orion Ophthalmology LLC Heterocyclic prolinamide derivatives

Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4668503A (en) * 1982-07-26 1987-05-26 Trustees Of University Of Massachusetts Process for labeling amines with 99m Tc
EP0293881A2 (fr) * 1987-06-05 1988-12-07 The Du Pont Merck Pharmaceutical Company Inhibiteurs peptides de l'acide boronique de proteases de type trypsine
EP0403243A1 (fr) * 1989-06-16 1990-12-19 Merck Frosst Canada Inc. Dérivés chélateurs du facteur natriurétique Atrial (ANF)
EP0471651A2 (fr) * 1990-08-13 1992-02-19 Sandoz Ltd. Dérivés de l'acide boronique peptidique ayant une activité inhibante de protéase
WO1992018536A2 (fr) * 1991-04-22 1992-10-29 Mallinckrodt Medical, Inc. Procede de detection et de localisation de tissus comprenant des recepteurs de neurokinine 1
US5252721A (en) * 1990-09-28 1993-10-12 Neorx Corporation S3 N chelating compounds

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4668503A (en) * 1982-07-26 1987-05-26 Trustees Of University Of Massachusetts Process for labeling amines with 99m Tc
EP0293881A2 (fr) * 1987-06-05 1988-12-07 The Du Pont Merck Pharmaceutical Company Inhibiteurs peptides de l'acide boronique de proteases de type trypsine
EP0403243A1 (fr) * 1989-06-16 1990-12-19 Merck Frosst Canada Inc. Dérivés chélateurs du facteur natriurétique Atrial (ANF)
EP0471651A2 (fr) * 1990-08-13 1992-02-19 Sandoz Ltd. Dérivés de l'acide boronique peptidique ayant une activité inhibante de protéase
US5252721A (en) * 1990-09-28 1993-10-12 Neorx Corporation S3 N chelating compounds
WO1992018536A2 (fr) * 1991-04-22 1992-10-29 Mallinckrodt Medical, Inc. Procede de detection et de localisation de tissus comprenant des recepteurs de neurokinine 1

Cited By (15)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1997005161A1 (fr) * 1995-07-28 1997-02-13 Novartis Ag Derives d'acides bores
WO2002022011A1 (fr) 2000-09-15 2002-03-21 Institut für Diagnostikforschung GmbH an der Freien Universität Berlin Procede de representation par images et de diagnostic de thrombus par tomographie a spin nucleaire faisant appel a des agents de contraste particulaires
EP1396269A1 (fr) 2002-09-09 2004-03-10 Trigen Limited Sels d'acide boronique des metaux multivalents et leur utilisation dans la préparation de médicaments pour le traitement de la thrombose
EP1466917A1 (fr) 2002-09-09 2004-10-13 Trigen Limited Procédé der préparation des acides boroniques peptidiques et acides obtenus
US7915236B2 (en) 2003-08-14 2011-03-29 Cephalon, Inc. Proteasome inhibitors and methods of using the same
US7576206B2 (en) 2003-08-14 2009-08-18 Cephalon, Inc. Proteasome inhibitors and methods of using the same
US7223745B2 (en) 2003-08-14 2007-05-29 Cephalon, Inc. Proteasome inhibitors and methods of using the same
US8058262B2 (en) 2003-08-14 2011-11-15 Cephalon, Inc. Proteasome inhibitors and methods of using the same
US8546608B2 (en) 2003-08-14 2013-10-01 Cephalon, Inc. Proteasome inhibitors and methods of using the same
US9233115B2 (en) 2003-08-14 2016-01-12 Millennium Pharmaceuticals Inc. Proteasome inhibitors and methods of using the same
US8283367B2 (en) 2005-02-11 2012-10-09 Cephalon, Inc. Proteasome inhibitors and methods of using the same
US8541590B2 (en) 2009-12-22 2013-09-24 Cephalon, Inc. Proteasome inhibitors and processes for their preparation, purification and use
US11267803B2 (en) 2016-06-21 2022-03-08 Orion Ophthalmology LLC Carbocyclic prolinamide derivatives
US11377439B2 (en) 2016-06-21 2022-07-05 Orion Ophthalmology LLC Heterocyclic prolinamide derivatives
US11866422B2 (en) 2016-06-21 2024-01-09 Orion Ophthalmology LLC Carbocyclic prolinamide derivatives

Also Published As

Publication number Publication date
IL112423A0 (en) 1995-03-30
SG47079A1 (en) 1998-03-20
AU1576495A (en) 1995-08-15
ZA95636B (en) 1996-07-26
PE45295A1 (es) 1995-12-29
GB9401483D0 (en) 1994-03-23
JPH09509653A (ja) 1997-09-30
CA2179889A1 (fr) 1995-08-03
EP0741747A1 (fr) 1996-11-13

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