Classifications

• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K49/00—Preparations for testing in vivo
  • A61K49/06—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations
  • A61K49/08—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by the carrier
  • A61K49/10—Organic compounds
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K49/00—Preparations for testing in vivo
  • A61K49/06—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations
  • A61K49/08—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by the carrier
  • A61K49/085—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by the carrier conjugated systems
• • 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
  • A61K51/0497—Organic compounds conjugates with a carrier being an organic compounds
• • 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
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K2121/00—Preparations for use in therapy
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K2123/00—Preparations for testing in vivo

Definitions

• the present invention relates to labelled biotin compounds, to a method of preparing these compounds, to a pharmaceutical composition comprising these compounds, to the use of this composition for diagnosis and therapy, and to a kit for preparing a radiopharmaceutical composition.
• MAb's monoclonal antibodies
• MAb*'s radiolabelled monoclonal antibodies
• (2nd step) and radiolabelled biotin (3rd step) are injected successively (in about 24 h time intervals) to the patient.
• the first two steps, as well as the first step in the two-step approach, result in the avidination of the tumour.
• the biotinylated MAb's which have not localized on the tumour but are still circulating in the blood, are macroaggregated by the excess of avidin.
• These avidin/MAb adducts of high molecular weight are rapidly taken up and catabolized by the liver.
• the 3rd step involves the in vivo administration of radiolabelled biotin. Due to the fast blood clearance of the non-tumour-bound biotin derivative, good tumour/background ratios are expected to be reached rapidly.
• biotin. labelled with a radionuclide preferably a metal-radionuclide, or with a paramagnetic metal ion are required. Since biotin is lacking Q is a group which is capable of reacting with an amino group
• the peptide and which is preferably selected from the group consisting of carbonyl, carbimidoyl, N-(C 1 -C 6 )alkylcarbimidoyl, N-hydroxycarbimidoyl and N-(C 1 -C 6 )alkoxycarbimldoyl.
• N 1 P q S (4-t-q) tetradentate chelating agents are selected from
• the above-defined spacing group Sp is preferably a group of the general formula
• A is a biradical of the formula
• n is an integer from 1 to 4.
• p is an integer from 0 to 4.
• X is O or S
• Y is NH, CO or S.
• R 1 is a branched or non-branched, optionally substituted
• biotin compounds of the invention can be prepared in a manner known per se for related compounds.
• the biotin molecule is derivatized with the desired chelating agent as defined hereinbefore, e.g. a N t P q S (4- ⁇ -q) tetradentate chelating agent, or EDTA
• n i or 2
• R is a chelating group for chelating a metal atom
• Sp is a spacing group having at least 4 atoms spacing NH from R;
• a metal as defined hereinbefore, in the form of a salt or of a chelate bound to a comparatively weak chelator, in order to form a complex.
• Suitable examples of salts or chelates of the desired metal are: 111 Irvcitrate and acetate, 99m Tc-tartrate, etc.
• the complex-forming reaction can generally be carried out in a simple manner and under moderate conditions.
• the invention also relates to a biotin compound of the general formula I, as defined above, which compound can be used for the above-described method of preparing the labelled biotin compound.
• the present invention further relates to a pharmaceutical composition, comprising in addition to a pharmaceutically acceptable carrier material and, if desired, at least one pharmaceutically acceptable adjuvant, as the active substance a labelled biotin compound as defined hereinbefore.
• the invention also relates to a method for detecting and localizing tumours in the body of a warm-blooded living being, which comprises (i) administering to said being a composition comprising (strept)avidin conjugated with a polypeptide having a selective affinity to said tumour, (ii) thereupon, after avidination of said tumour, administering to wherein:
• R 6 -R 20 are each individually hydrogen atoms or (C 1 -C 4 )alkyl groups, with the proviso that at least one of C 6 to C 9 is the symbol Y;
• R 21 is a hydrogen atom or a CO 2 (C 1 -C 4 )alkyl group
• R 22 and R 23 are each individually (C 1 -C 4 )alkylcarbonyl, benzoyl or benzyl groups;
• v 0 or 1
• s 2 or 3;
• R 24 is CH 2 COOH or a functional derivative thereof
• A is (C 1 -C 4 )alkylene, if desired substituted with CO 2 alkyl, CH 2 COalkyl, CONH 2 ,
• G is NH or S
• Y is a valence bond or a functional group capable of binding with the spacing group
• Y preferably comprises isocyanato, isothiocyanato, formyl, o-halonitrophenyl, diazonium, epoxy, trichloro-s-triazinyl, ethyleneimino, chlorosulfonyl, alkoxycarbimidoyl, (substituted or unsubstituted) alkylcarbonyloxycarbonyl, alkylcarbonylimidazolyl, succinimido-oxycarbonyl, which group is prferably attached to a
• hydrocarbon biradicals are biradicals derived from benzene, (C 1 -C 6 )alkanes, (C 2 -C 6 )alkenes and (C 1 -C 4 )alkyibenzenes.
• suitable chelators of the general formula III are described in the international patent application WO 89/07456, such as unsubstituted or substituted 2-iminothiolanes and 2-iminothiacyclohexanes, in particular 2-imino-4-mercaptomethytthiolane.
• biotin compounds have been tested in a number of suitable model experiments that are predictive for in vivo application. These experiments are described in the Examples. From the results of these experiments it will be evident, that the labelled biotin compounds of the present invention have properties which make them suitable for diagnostic and therapeutic purposes. If labelled with a suitable atom for diagnostic purposes, the labelled biotin compound remains sufficientiy long intact after administration to permit imaging of the target tumour without presenting a disturbing background. If labelled with a suitable radioisotope for therapy, such-labelled biotin compounds are promising therapeutic agents for the treatment of a number of malignant tumours.
• the body of ⁇ warm-blooded living being which comprises (i) administering to said being a composition comprising (strept)avidin conjugated with a polypeptide having a selective affinity to said tumour, (ii) thereupon, after avidination of said tumour, administering to said being a composition comprising,, in a quantity effective for combating or controling tumours, a labelled biotin compound as defined hereinbefore, said biotin compound being labelled with a metal isotope selected from the group consisting of 114m In.
• kit may comprise (a) a composition comprising (strept)avidin conjugated with a polypeptide having a selective affinity to tumours, (b) a biotine compound of the general formula I, shown above, wherein the symbols have the above meanings, to which compound, if desired, an inert pharmaceutically acceptable carrier and/or formulating agents and/or adjuvants is/are added, (c) a solution of a salt or chelate of a metal isotope selected from the group consisting of 203 Pd, 66 Ga. 67 Ga, 68 Ga 72 As, 111 In, 113m In.
• the biotin compound to be used as an ingredient of the above kit has been modified by a spacing group Sp and a chelating agent R as defined hereinbefore.
• the resulting biotin compound provides a facility for firmly attaching the radionuclide in a simple manner.
• Suitable chelating agents for modifying the biotin molecule are said being a composition comprising, in a quantity sufficient for external imaging, a labelled biotin compound as defined hereinbefore, said biotin compound being labelled with (a) a radioactive metal isotope selected from the group consisting of 99m Tn, 203 Pd, 67 Ga, 68 Ga, 72 As, 111 In, 113m ln, 97 Ru, 62 Ga, 64 Ga, 52 Fe, 52m Mn and 51 Cr, or (b) with a paramagnetic metal atom selected from the group consisting of Cr, Mn, Fe, Co, Ni, Cu,
• the three-step pretargeting method can be used as discussed hereinbefore.
• the following protocol is used: (i- ⁇ ) administering to said being a composition comprising a biotinylated polypeptide having a selective affinity to said tumour, (i-b) then administering a composition comprising (strept)avidin; both steps in substitution for the above step (i).
• the invention also relates to a method of intraoperatively detecting and localizing tumours in the body of a warm-blooded living being, which comprises (i) administering to said being a composition comprising (strept)avidin conjugated with a polypeptide having a selective affinity to said tumour, (ii) thereupon, after avidination of said tumour, administering to said being a composition comprising, in a quantity sufficient for detection by a gamma detecting probe, a labelled biotin compound as defined hereinbefore, said biotin compound being labelled with 161 Tb, and (iii) finally, after allowing the active substance to be bound by and taken up in said tumour and after blood clearance of radioactivity, subjecting said being to a radioimmunodetection technique in the relevant area of the body of said being, by using a gamma detecting probe.
• the above radioisotope allows the use of a such-labelled peptide compound in the technique of radioguided surgery, wherein relevant tissues in the body of a patient can be detected and located intraoperatively by means of a gamma detecting probe.
• the surgeon can, intraoperatively, use this probe to find the lesions in which uptake of the compound labelled with said radioisotope, which is a low-energy gamma photon emittor, has taken place.
• the three-step pretargeting method can be used as discussed hereinbefore.
• the biotin compounds of the present invention provided they are radiolabelled with isotopes suitable for such purpose, can be used for the therapeutic treatment of tumours. So the invention further relates to a method for the therapeutic treatment of tumours in excellently suitable.
• suitable Sn(II)-cornpounds are SnCl 2 , Sn(II)-tartrate, Sn(II)-phosphonate or -pyrophosphate, and Sn(II)-glucoheptonate.
• the modified biotin constituent of the above-mentioned kits i.e.
• the biotin compound may be supplied as a solution, for example, in the form of a physiological saline solution, or in some buffer solution, or may be present in a dry condition, for example, in a lyophilized condition.
• a component for an injection liquid it should be sterile, in which, when the constituent is in the dry state, the user should preferably use a sterile physiological saline solution as a solvent.
• the abovementioned constituent may be stabilized in the conventional manner with suitable stabilizers, for example, ascorbic acid, gentisic acid or salts of these acids, or it may comprise other auxiliary agents, for example, fillers, such as glucose, lactose, mannitol, and the like.
• N t P q S (4-t-q) tetradentate chelating agents or N-containing di- or polyacetic acids or their derivatives, such as the compounds mentioned before, have proved to be pre-eminently suitable for attaching various metal radionuclides, such as ln-111 and ln-113m, to the biotin molecule.
• the Wt to be supplied to the user may also comprise the ingredient(s) defined sub (a) and (b) above, together with instructions for use, whereas the solution of a salt or chelate of the radionuclide, defined sub (c) above, which solution has a limited shelf life, may be put to the disposal of the user separately.
• Wt is destined for use in a two-step protocol. If, instead thereof, the three-step pretargeting method is to be used, as discussed hereinbefore, the following Wt is suitable:
• kits serve to prepare a radiopharmaceutical composition labelled with Tc- 99m, Re-186 or Re-188
• a Wt may comprise, in addition to the ingredient(s) defined sub (a) and (b) above, (c) a reducing agent and, if desired, a chelator, and (d) instructions for use with a prescription for reacting the ingredients of the Wt with Tc-99m in the form of a pertechnetate solution, or with Re-186 or Re-188 in the form of a perrhenate solution.
• certain ingredients of the kit may be combined, provided they are compatible.
• the pertechnetate or perrhenate solution can simply be obtained by the user from a suitable generator.
• the complex forming reaction with the biotin compound of the general formula I above can simply be produced by combining the components in a neutral or buffered medium and causing them to react.
• the radionuclide may be presented to said biotin compound in the form of a chelate bound to a comparatively weak chelator, as described hereinbefore.
• the Wt comprises a biotin compound as defined hereinbefore and is intended for the preparation of a radiopharmaceutical composition, labelled with Tc-99m, Re-186 or Re-188
• the radionuclide will preferably be added separately in the form of a pertechnetate or perrhenate solution.
• the kit will comprise a suitable reducing agent and, if desired, a chelator, the former to reduce the pertechnetate or the perrhenate.
• a suitable reducing agent may be used, for example, a dithionite or a metallic reducing agent.
• a metallic reducing agent for example, Sn(II), Ce(III), Fe(II), Cu(I), Ti(III) or Sb(III);
• 6-(p-Nitrobenzyl)-1,4,8,11-tetraazaundecane (2) The crude (1) (480 mg, 1 ,48 mmol) is reacted at RT with 1 M BH 3 /THF solution (30 ml, 20 eq) and the reaction mixture is refluxed for 60 h.
• MeOH is added dropwise with caution to the reaction mixture, which is precooied at 0° C.
• the MeOH is evaporated in vacuum and the residue is suspended in EtOH and sonicated for several minutes.
• the undissolved part is filtered off and HCl gas is introduced for 20 min to the filtrate.
• the formed precipitate is collected and further purified by column chromatography (SiO 2 ;CHCl 3 /MeOH/25% NH 3 5/5/2).
• the resulting purified amine is dissolved in EtOH and the corresponding hydrochloric salt is obtained as a white solid by passing HCl gas to this solution (460 mg).
• the precipitate is directly recrystallized from HCl gas saturated MeOH, without previous purification by chromatography.
• Biotinsulfoxide is synthesized according to Melville (D.B. Melville, J. Biol. Chem., 1954, 208, 495).
• the sulfoxide exists in two isomeric forms: ⁇ -(+)-biotinsulfoxide and ß-(-)-biotinsulfoxide (Fig.2)
• Biotinidase is an enzyme found in different organs and in human serum which functions as a biotin-amide amidohydrolase; e.g. it hydrolyzes (N-(d-biotinyl)-p-aminobenzoic acid and biocytin.
• the stability of the amide bond in fresh human serum is studied. Within 4 h of incubation the amount of intact ( 99m Tc)-(6), obtained according to Example II, decreases by about 15 ⁇ 2% which is by far superior to the corresponding conjugate with biotin.
• the optimal time of scintigraphy with a 99m Tc-labelled biotin conjugate is within 2 h post injection, the stability of 99m Tc(6) in human serum is adequate for human studies.
• EDTA ethylene diamine tetra-acetic acid
• DTPA diethylene triamine penta
• TTA and TRITA hydroxyethyldiamine triacetic acid
• HEDTA hydroxyethyldiamine triacetic acid
• TETA 1,4,8,11-tetra-azacyclotetradecane-N,N',N",N'"-tetra-acetic acid
• substituted DTPA substituted EDTA or from deferoxamine or from a compound of the general formula
• R 1 is a branched or non-branched, optionally substituted
• hydrocarbyl radical which may be interrupted by one or more hetero-atoms selected from N, O and S and/or by one or more NH groups, and
• Q is a group which is capable of reacting with an amino group
• peptide which is preferably selected from the group consisting of carbonyl, carbimidoyl, N-(C 1 -C 6 )alkylcarbimidoyl, N-hydroxycarbimidoyl and

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• 1996
  • 1996-09-17 JP JP9512833A patent/JPH11513374A/ja active Pending
  • 1996-09-17 EP EP96933801A patent/EP0857071A1/de not_active Withdrawn
  • 1996-09-17 CZ CZ98754A patent/CZ75498A3/cs unknown
  • 1996-09-17 WO PCT/US1996/014883 patent/WO1997010854A1/en not_active Application Discontinuation
  • 1996-09-17 CA CA002232411A patent/CA2232411A1/en not_active Abandoned
• 1998
  • 1998-03-17 NO NO981202A patent/NO981202L/no unknown
  • 1998-03-18 IL IL12372598A patent/IL123725A0/xx unknown

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