WO2001015746A1 - Radiopharmaceutical products and their preparation procedure - Google Patents

Radiopharmaceutical products and their preparation procedure Download PDF

Info

Publication number
WO2001015746A1
WO2001015746A1 PCT/IB2000/001161 IB0001161W WO0115746A1 WO 2001015746 A1 WO2001015746 A1 WO 2001015746A1 IB 0001161 W IB0001161 W IB 0001161W WO 0115746 A1 WO0115746 A1 WO 0115746A1
Authority
WO
WIPO (PCT)
Prior art keywords
polysaccharide
groups
sequestering
starch
product according
Prior art date
Application number
PCT/IB2000/001161
Other languages
French (fr)
Inventor
Emmanuel Bellande
Pierre Jallet
Benoît DENIZOT
Original Assignee
Schering Aktiengesellschaft
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
Priority to JP2001520157A priority Critical patent/JP2003508455A/en
Priority to NZ517377A priority patent/NZ517377A/en
Application filed by Schering Aktiengesellschaft filed Critical Schering Aktiengesellschaft
Priority to PL00353803A priority patent/PL353803A1/en
Priority to CA002383517A priority patent/CA2383517A1/en
Priority to EA200200307A priority patent/EA200200307A1/en
Priority to EP00951784A priority patent/EP1210127A1/en
Priority to IL14807600A priority patent/IL148076A0/en
Priority to MXPA02001923A priority patent/MXPA02001923A/en
Priority to HU0202714A priority patent/HUP0202714A2/en
Priority to SK277-2002A priority patent/SK2772002A3/en
Priority to KR1020027002820A priority patent/KR20020040799A/en
Priority to BR0013729-4A priority patent/BR0013729A/en
Priority to AU64631/00A priority patent/AU6463100A/en
Priority to EEP200200105A priority patent/EE200200105A/en
Publication of WO2001015746A1 publication Critical patent/WO2001015746A1/en
Priority to BG06438A priority patent/BG106438A/en
Priority to NO20021001A priority patent/NO20021001L/en
Priority to HK02106423.2A priority patent/HK1044893A1/en

Links

Classifications

    • 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
    • A61K51/12—Preparations containing radioactive substances for use in therapy or testing in vivo characterised by a special physical form, e.g. emulsion, microcapsules, liposomes, characterized by a special physical form, e.g. emulsions, dispersions, microcapsules
    • 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
    • A61K51/06—Macromolecular compounds, carriers being organic macromolecular compounds, i.e. organic oligomeric, polymeric, dendrimeric molecules
    • A61K51/065—Macromolecular compounds, carriers being organic macromolecular compounds, i.e. organic oligomeric, polymeric, dendrimeric molecules conjugates with carriers being macromolecules
    • 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
    • A61K51/12—Preparations containing radioactive substances for use in therapy or testing in vivo characterised by a special physical form, e.g. emulsion, microcapsules, liposomes, characterized by a special physical form, e.g. emulsions, dispersions, microcapsules
    • A61K51/1241—Preparations containing radioactive substances for use in therapy or testing in vivo characterised by a special physical form, e.g. emulsion, microcapsules, liposomes, characterized by a special physical form, e.g. emulsions, dispersions, microcapsules particles, powders, lyophilizates, adsorbates, e.g. polymers or resins for adsorption or ion-exchange resins
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents

Definitions

  • the present invention relates to radiopharmaceutical products which can be used for diagnosis or therapy and to their preparation procedure .
  • radiopharmaceutical products formed for example from a suspension of particles labelled by a radioactive isotope utilisable in particular for pulmonary scintigraphy, for example in order to establish a diagnosis when a pulmonary embolism is suspected.
  • the products are used under the form of particles which are preferably spherical in shape and of a size ranging from 10 to lOO ⁇ m.
  • the pulmonary capillaries have a diameter of about 7 ⁇ m, the particles remain blocked in the capillaries after their intravenous injection, which makes it possible to visualise anomalies of pulmonary blood perfusion.
  • these products must fulfil a certain number of pharmaceutical restrictions.
  • they must have a suitable degradation rate in vivo, that is sufficiently slow to allow imagery to be carried out, for example by a gamma-ray camera, a minimum of about one hour, but also sufficiently rapid so as not to provoke permanent obstruction of the pulmonary capillaries, which could give rise to small thromboses.
  • these products must not be toxic for the organism, they must be able to be sterilised for example by autoclaving or by irradiation, they must be able to be labelled easily with a radioactive metal and be able to be packaged under the form of a stable labelling kit.
  • the inventors demonstrated other defects of these microparticles in the comparative examples 1 and 2 below.
  • the particles comprise chelating groups linked by covalent bonds to which the radioactive nucleus is linked under the form of chelate type complexes which are principally composed of at least four, and preferably at least five to eight cyclic nuclei with 5 to 6 groups, enclosing the metal, and two metal-coordinating atoms.
  • the polysaccharide is a polysaccharide reticulated chemically, for example by means of epichlorhydrin or epibromhydrin.
  • the labelling procedure comprises heating to 100°C in the presence of the radioactive element, a washing and a drying after labelling, which is not at all compatible with the idea of the above- mentioned labelling kit and the restrictions of sterility of usage.
  • the labelling method described allows the particles to be labelled in a relatively stable manner, it does not make it possible to prepare a labelling kit which is pharmaceutically acceptable, in particular because it contains epichlorhydrin, and easily usable in a nuclear medicine service.
  • microspheres described in these two brevet applications are thus not adapted to the pharmaceutical restrictions and they cannot be exploited. Moreover they have never been used for pulmonary scintigraphy. This type of product has been abandoned since.
  • radioactive particles intended for the study of pulmonary perfusion.
  • the particles described in this document are macro-aggregates of radio- iodinated serum albumin ( 131 I-MAA) or microspheres of denatured human serum albumin labelled with technetium
  • the microspheres of 99m Tc-HAM are preferable, because of their uniformity of particle size ranging essentially between 40 and 50 urn. Moreover this document describes the general characteristics required for such radiopharmaceutical particles.
  • the precise aim of the invention is to overcome the inconveniences mentioned above for prior art products, by providing a radiopharmaceutical product being able to be easily labelled, for example with 99m Tc, presenting a very good pulmonary captation which has been demonstrated by inventors for rats, non-toxic, easily biodegradable, easily sterilisable and able to be packaged as a kit ready for labelling, stable and fulfilling the pharmaceutical restrictions for this type of product.
  • the utilisable alkyl groups for R' can be linear or branched, and preferably they have 1 to 5 carbon atoms .
  • the polysaccharide can be soluble, or in the form of microparticles.
  • the polysaccharide can be chosen, for example, from among natural starch, cellulose or reticulated amyl pectin.
  • the natural starch can, for example, be maize starch.
  • the polysaccharide can be in the form of microparticles, for example in the form of microspheres .
  • modified cellulose according to the present invention offers very good pulmonary captation and an elimination speed slower than with starch.
  • the modified cellulose of the present invention can therefore also be used for radiotherapy, for example with labelling with rhenium, copper, or with one of the above-mentioned metals, since it corresponds to the radiotherapy necessity of using microparticles with a longer half-life.
  • the sequestering groups can be chosen for example from the groups with formulae:
  • R 1 , R 2 , R 3 , R 4 and R 5 are independently hydrogen atoms , saturated or unsaturated hydrocarbonic groups , carboxylic groups or aromatic groups,
  • n is an aromatic nucleus possibly containing one or several hetero-atoms, and n is a whole number between 1 and 5.
  • the microparticles for example in the shape of microspheres, can be of a dimension from 0.01 to 100 ⁇ m, preferably from 10 to 50 ⁇ m for diagnosis by pulmonary scintigraphy and from 0.1 to 5 ⁇ m for therapy.
  • the levels of sequestering groups can be from 0.1 to 50% compared to the saccharide patterns of the polysaccharide, preferably from 2 to 15%.
  • the radioactive metal in the radiopharmaceutical product, particularly when it is used for diagnosis, can be 99 Tc or gallium-67.
  • the radioactive metal in the radiopharmaceutical product, in particular when it is used for therapy, can be rhenium-
  • said radiopharmaceutical product can be in the form of a suspension of microspheres in a physiologically acceptable liquid or in lyophilised form.
  • the present invention also provides a preparation procedure for the radiopharmaceutical product of the invention comprising the following stages:
  • a radioactive metal chosen from among technetium, rhenium, copper, yttrium, erbium and samarium.
  • the oxidation controlled by means of a periodate can be that described, for example, in CL. Mehltretter, "Methods in Carbohydrate Chemistry", vol. IV, 1964, applied in particular to the oxidation of starch, dextrane or cellulose. It is used in the examples given below.
  • the compound containing a primary amine function can correspond to the formula NH 2 - (CH 2 ) n -SH with n being a whole number between 1 and 5, and can include a supplementary reducing stage of this compound with sodium borohydride between stages (b) and (c) .
  • the compound bonded to the polysaccharide can correspond, for example, to one of the following formulae:
  • the level of sequestering groups fixed on the polysaccharide can be regulated by controlling the level of oxidation of the polysaccharide in stage (a) mentioned above.
  • This oxidation level of polysaccharide can, for example, be between 10 to 50%.
  • the level of the sequestering group can, for example, be between 2 and 15%.
  • the inventors have therefore used a two-stage transformation method.
  • This method can be presented as consisting of carrying out a controlled oxidation of the polysaccharide by the periodate in the first stage.
  • Each unit of oxidised glucose thus generates two aldehyde groups in neighbouring positions following the chemical reaction diagram given below:
  • the level of oxidation of the polysaccharide can be variable and easily adjusted. In fact, the yield of this oxidation reaction is close to 100% and the level of oxidaton can be calculated from the quantities of periodate added. In general, oxidation levels lower than 50% are used so as to modify the structure of the macromolecule only slightly. The real oxidation level, ranging from 1 to 100%, can easily be determined by a colorometric method.
  • the oxidised polysaccharide is made to react with a molecule containing an amine or hydrazin grouping with the general formula RNH 2 or RNHNH 2 to form a chelating grouping able to sequester technetium.
  • a Schiff base type ligand or thiosemicarbazone is obtained.
  • This second stage can be summarised as follows: 0
  • stage (c) can for example consist of putting into contact the microspheres of polysaccharide comprising the sequestering groups for example with a solution of pertechnetate 99m Tc0 4 " in the presence of a reducing agent, for example stannous chloride.
  • microparticles for example microspheres, for example maize starch or starch with a base of reticulated amyl pectin can thus be oxidised, then coupled to a molecule containing an amine or hydrazin function, for example S-methyl dithiocarbazate.
  • a molecule containing an amine or hydrazin function for example S-methyl dithiocarbazate.
  • the present invention thus provides in particular microparticles prepared for example from a base of starch particles, which therefore do not present the inconveniences of the albumin mentioned above.
  • the starch is described as an excipient in the pharmacopoeia. It is therefore easily available and at low cost.
  • microparticles of the present invention also have the advantage of being able to be sterilised easily, for example by irradiation, and to be processed under the form of a kit ready for labelling.
  • the present inventors have demonstrated according to the present invention that the speed of pulmonary clearance can be modified according to the level of oxidation of the microparticles used in the present invention, which is not possible, for example, with human albumin microspheres.
  • Another advantage of the present invention lies in the simplicity of operation of the procedure: the reaction conditions being very gentle: reactions at ambient temperature, in an aqueous medium, quasi- quantitative yields.
  • the sequestering reactions for example with technetium, are quantitative; they take place at room temperature and without final purification which makes it possible to adapt to the requirements of sterility and simplicity of preparation necessary for the utilisation of technetised kits in the hospital environment.
  • the present invention also provides a diagnosis kit which can be used, for example, for pulmonary scintigraphy.
  • R' in which R is a hydrocarbonic or aromatic group comprising at least one atom of sulphur, and in which R' is a hydrogen atom or an alkyl grouping such as methyl.
  • the polysaccharide can, for example, be in the form of microparticles, for example in the shape of microspheres, the microparticles being able to be in lyophilised form or in suspension in a pharmaceutically acceptable liquid.
  • the kit of the present invention can furthermore comprise a second flask containing stannous chloride preferably uin lyophilised form, or also when the polysaccharide is in lyophilised form, for example in the form of microparticles, in the first flask, said first flask can besides contain lyophilised stannous chloride.
  • the kits of the present invention are stable for at least twelve months as demonstrated in the examples given below.
  • the radiopharmaceutical product of the present invention therefore presents all the qualities required for a use such as radiopharmaceutical usage, for example for scintigraphy of pulmonary perfusion or for radiotherapy.
  • a suspension of 10 g of maize starch from the pharmacopoeia is prepared, previously sieved between 10 and 40 ⁇ m, containing about 10% water, that is 0.055 mole of glucose in 100 ml of water.
  • the suspension is then stirred for 18 hours at room temperature.
  • the solution is filtered and the oxidised starch is rinsed by 5 times 20 ml of water and then 2 times 50 ml of acetone.
  • the assay of the powder by elementary analysis gives a sulphur content of 5.4%, which corresponds to a coupling level of S-methyl dithiocarbazate (DTCZ) of 7% (7 units dithiocarbazate for 100 theoretical aldehyde functions that is 14 dithiocarbazate units for 100 glucose units) .
  • the coupling yield is therefore 70%.
  • the assay of the powder by elementary analysis gives a sulphur content of 5%, which corresponds to a coupling level of the N- methyl S-methyl dithiocarbazate of 6.5% (6.5 units dithiocarbazate for 100 theoretical aldehyde functions, that is 13 dithiocarbazate units for 100 units glucose).
  • the coupling yield is thus 65%.
  • the assay of the powder by elementary analysis gives a sulphur content of 3%, which corresponds to a coupling level of the 4,4-dimethyl 3-thiosemicarbazone of 7.5% (7.5 units thiosemicarbazide for 100 theoretical aldehyde functions, that is 15 thiosemicarbazone units for 100 units glucose).
  • the coupling yield is thus 75%.
  • the assay of the powder by elementary analysis gives a sulphur content of 3%, which corresponds to a coupling level of the 4-allyl 3-thiosemicarbazone of 7.5% (7.5 units thiosemicarbazide for 100 theoretical aldehyde functions, that is 15 thiosemicarbazone units for 100 units glucose).
  • the coupling yield is thus 75%.
  • Starch modification One proceeds as for example 1 to obtain 10 g of starch oxidised at 30%.
  • a suspension is prepared of 10 g of starch oxidised at 30% in 60 ml of a mixture of water/ethanol (2/1 by volume).
  • 0.1 eq 0.1x0.055x2
  • 0.1 eq 0.1x0.055x2
  • the suspension is stirred for 18 hours at room temperature.
  • the solution is then filtered and the modified starch washed by 3 times 20 ml ethanol and then vacuum dried.
  • the assay of the powder by elementary analysis gives a sulphur content of 2.9%, which corresponds to a coupling level of the 3-thiosemicarbazone of 7.3% (7.3 units thiosemicarbazide for 100 theoretical aldehyde functions, that is 14.6 thiosemicarbazone units for 100 units glucose).
  • the coupling yield is thus 73%.
  • the assay of the powder by elementary analysis gives a sulphur content of 3.4%, which corresponds to a coupling level of the 2-mercaptoethylamine of 8.5% (8.5 units 2-mercaptoethylamine for 100 theoretical aldehyde functions, that is 17 2-mercaptoethylamine units for 100 units glucose).
  • the coupling yield is thus 85%.
  • the assay of the powder by elementary analysis gives a sulphur content of 2.8%, which corresponds to a coupling level of the 2-amino 4-mercaptotriazole of 7% (7 units mercaptotriazole for 100 theoretical aldehyde functions, that is 14 mercaptotriazole units for 100 units glucose) .
  • the coupling yield is thus 75%.
  • Comparative example 1 One uses 10 g of sieved pharmacopoeia maize which has not undergone chemical transformation and one caries out the same procedure as in example 1 to label it with 99m Tc.
  • the RCP is 19%.
  • Example 14 One proceeds as in example 1.
  • the RCP is 99.1%.
  • Example 14 One proceeds as in example 1.
  • the RCP is 99.1%.
  • Sprague Dawley rats weighing about 200 g are anaesthetised with sodium thiopental, and are injected intravenously with different solutions of microspheres labelled with 99 Tc according to examples 1 to 9 and 13.
  • Each animal receives 0.2 ml of solution in the penis vein, that is 0.2 mc per animal.
  • the animals are then placed under a gamma-ray camera and successive static images are shot over a period of 3 hours. One thus obtains successive images after acquiring 15,000 shots per image. Then, manually, one defines the zones of interest in order to estimate the activity present in the different organs 15 minutes after the injection. The results are given in tables I below.
  • modified microspheres show very good pulmonary captation.
  • microspheres of reticulated amyl pectin not modified chemically are labelled by 99m Tc but do not present any pulmonary captation, doubtless due to the weak link between 99 Tc and the microspheres.
  • these microspheres transformed chemically by the procedure of the invention demonstrate good pulmonary captation.
  • Starch microspheres prepared as in example 1 (starch oxidised at 30%, coupled with DTCZ at 7%) are used to produce sterile labelling kits and are ready for labelling with 99m Tc . Sterilisation of the microspheres
  • microspheres 10 g are introduced into a flask crimped and then irradiated by a source of cobalt-60.
  • the microspheres receive a total dose of gamma radiation of 25kGy over 20 hours.
  • 200 mg of sterilised microspheres are introduced in a sterile way into a reactor containing 20 ml of NaCl 9/1000.
  • the solution is de-aerated by nitrogen bubbling and then one adds 400 ⁇ l of a sterile solution of SnCl 2 , 2H 2 0 at 0.5 mg/ml in HCI 0.1N.
  • Each flask thus contains :
  • Sprague Dawley rats weighing about 200 g are anaesthetised with sodium thiopental, and are then injected intravenously with different solutions of microspheres labelled with 99m Tc as in example 14. Each animal receives 0.2 ml of solution in the penis vein, that is 0.2 mc per animal. The animals are then killed 15 minutes after the injection. Next, the different organs are retrieved, the measurement of radioactivity present in each organ is counted and thus the percentage of activity present in each organ is determined. The results are given in table IV below:
  • HSA human serum albumin
  • Sprague Dawley rats weighing about 200 g are anaesthetised and then injected with 0.2 ml (0.1 mc) of cellulose microsphere solution labelled with Re 186 as in example 18.
  • the animals are then placed under a gamma-ray camera and images are registered over 48 hours.
  • the activity present in the zones of interest is then calculated as in example 14.
  • liver cancers after injection, not intravenously but directly into the hepatic artery (metabolic radiotherapy) .
  • Another possible application is to inject this type of particle subcutaneously in breast cancer.
  • the particles migrating through the lymphatic system may make it possible to treat the sentinel nodes invaded by cancerous cells .

Landscapes

  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • Veterinary Medicine (AREA)
  • Medicinal Chemistry (AREA)
  • Public Health (AREA)
  • General Health & Medical Sciences (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Optics & Photonics (AREA)
  • Physics & Mathematics (AREA)
  • Epidemiology (AREA)
  • Dispersion Chemistry (AREA)
  • Organic Chemistry (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • General Chemical & Material Sciences (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
  • Medicinal Preparation (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)

Abstract

The present invention relates to radiopharmaceutical products and their preparation procedure. These products can be used for pulmonary scintigraphy or for therapy. They comprise a polysaccharide and sequestering groups of formulae R-NH-, R-N=, and formula (I) in which R is a hydrocarbonic or aromatic group comprising at least one atom of sulphur, and R' is an atom of hydrogen or an alkyl grouping such as methyl, said sequestering groups forming a chelate type complex with a radioactive metal such as technetium.

Description

RADIOPHARMACEUTICAL PRODUCTS AND THEIR PREPARATION PROCEDURE
Technical field
The present invention relates to radiopharmaceutical products which can be used for diagnosis or therapy and to their preparation procedure .
In particular, it relates to radiopharmaceutical products formed for example from a suspension of particles labelled by a radioactive isotope utilisable in particular for pulmonary scintigraphy, for example in order to establish a diagnosis when a pulmonary embolism is suspected.
In this application, the products are used under the form of particles which are preferably spherical in shape and of a size ranging from 10 to lOOμm. In fact, since the pulmonary capillaries have a diameter of about 7μm, the particles remain blocked in the capillaries after their intravenous injection, which makes it possible to visualise anomalies of pulmonary blood perfusion.
Evidently these products must fulfil a certain number of pharmaceutical restrictions. In particular they must have a suitable degradation rate in vivo, that is sufficiently slow to allow imagery to be carried out, for example by a gamma-ray camera, a minimum of about one hour, but also sufficiently rapid so as not to provoke permanent obstruction of the pulmonary capillaries, which could give rise to small thromboses. In addition, these products must not be toxic for the organism, they must be able to be sterilised for example by autoclaving or by irradiation, they must be able to be labelled easily with a radioactive metal and be able to be packaged under the form of a stable labelling kit.
Prior Art For example, the application for French brevet FR- A-2 273 516, deposited in 1975 by the PHARMACIA AKTIEBOLAG Company, resident in Sweden, describes the use of microspheres of amyl pectin reticulated by epichlorhydrin and labelled by a simple mixture with 99Tc for pulmonary perfusion scintigraphy . These particles present several inconveniences. In fact, only the hydroxyl groupings of amyl pectin used can allow this mixture labelling, and unfortunately they only form weak bonds with technetium and do not make stable labelling possible. In addition, the preparation procedure described uses many solvents and emulsifiers which are difficult to eliminate from the particles prepared. Furthermore, the exact rate of reticulation cannot be measured accurately nor controlled on this particle type.
Moreover, this document does not describe the kit compatible with routine utilisation in nuclear medicine. In fact, for an injectable preparation for humans, several manipulations such as adjunction of tin to the sterile flask, a centrifuging, a restoration of suspension, etc. are necessary, which is not compatible with sterility requirements. Finally, the solutions obtained are not stable and the epichlorhydrin used for reticulation is recognised as being very toxic and mutagenic .
The inventors demonstrated other defects of these microparticles in the comparative examples 1 and 2 below.
The application for French brevet FR-A-2 285 857 deposited in 1975 by the PHARMAGIA FINE CHEMICALS AB Company, resident in Sweden, describes the utilisation of polysaccharide particles linked to different sequestering agents and labelled with the aid of radioactive isotopes. The particles comprise chelating groups linked by covalent bonds to which the radioactive nucleus is linked under the form of chelate type complexes which are principally composed of at least four, and preferably at least five to eight cyclic nuclei with 5 to 6 groups, enclosing the metal, and two metal-coordinating atoms. The polysaccharide is a polysaccharide reticulated chemically, for example by means of epichlorhydrin or epibromhydrin. Leaving the labelling aside, these particles present the same problems as those mentioned previously for the particles described in FR-A-2 273 516. Moreover, this document does not give any examples of labelling with technetium. Further, the labelling procedure comprises heating to 100°C in the presence of the radioactive element, a washing and a drying after labelling, which is not at all compatible with the idea of the above- mentioned labelling kit and the restrictions of sterility of usage. Even though the labelling method described allows the particles to be labelled in a relatively stable manner, it does not make it possible to prepare a labelling kit which is pharmaceutically acceptable, in particular because it contains epichlorhydrin, and easily usable in a nuclear medicine service.
The microspheres described in these two brevet applications are thus not adapted to the pharmaceutical restrictions and they cannot be exploited. Moreover they have never been used for pulmonary scintigraphy. This type of product has been abandoned since.
The many researches carried out since 1975 for perfecting new radiopharmaceutical products have concentrated on products based on albumin-serum and its derivatives. These blood products do in fact correspond to pharmaceutical restrictions and can be used in particular for pulmonary scintigraphy. These are the products used at present in nuclear medicine.
For example, in 1975, M.A. Davis, in the document "Radiopharmaceuticals N.Y.", 1975, pages 267 to 281, described the radioactive particles intended for the study of pulmonary perfusion. The particles described in this document are macro-aggregates of radio- iodinated serum albumin (131I-MAA) or microspheres of denatured human serum albumin labelled with technetium
(99mTc-HAM) . The microspheres of 99mTc-HAM are preferable, because of their uniformity of particle size ranging essentially between 40 and 50 urn. Moreover this document describes the general characteristics required for such radiopharmaceutical particles. The document of R. Guiraud "Macro-aggregates and radioactive microspheres", Radiopharmaceuticals , 1997, 519, describes macro-aggregates of albumin (MAA) and microspheres of human serum albumin. It describes the labelling of such micro-aggregates and microparticles with technetium 99m by a solution of stannous chloride. It also notes that the optimum size for the microparticles is 15±5 um. It mentions organic microspheres of starch. At present, these macro-aggregates and microspheres of human serum albumin labelled with 99mTc are by far the most utilised in nuclear medicine. However, they present several inconveniences. For example, the variability and quality of batches of human albumin sometimes make preparation of diagnosis kits difficult, containing particles which can vary in size and number. But one of the major inconveniences is their human origin, which can pose serious problems of potential vital contamination of the type HIV, hepatitis, or Creutzfeld-Jacob disease.
It would therefore be very interesting to be able to have microspheres labelled with 99mTc which are not of human origin in order to ensure perfect safety.
With this in view, the very recent document of A.C. Perkins, Nuclear Medicine Communications, 1999, 20, 1-3 describes ways of replacing radiopharmaceutical products obtained from blood. In particular it mentions the utilisation of recombinant materials, synthetic polymers and polypeptides. But, this document does not mention polysaccharides . Description of the invention
The precise aim of the invention is to overcome the inconveniences mentioned above for prior art products, by providing a radiopharmaceutical product being able to be easily labelled, for example with 99mTc, presenting a very good pulmonary captation which has been demonstrated by inventors for rats, non-toxic, easily biodegradable, easily sterilisable and able to be packaged as a kit ready for labelling, stable and fulfilling the pharmaceutical restrictions for this type of product. These advantages and others will be evident from the following description.
The radiopharmaceutical product of the present invention is characterised in that it comprises a polysaccharide provided with sequestering agents linked to the polysaccharide by covalent bonds and chosen among the groups of formulae R-NH- , R-N= , and R N N =
R' in which R is a hydrocarbonic or aromatic group comprising at least one atom of sulphur, and R' is a hydrogen atom or an alkyl grouping, for example methyl, said sequestering groups forming a chelate type complex with a radioactive metal chosen from among technetium, rhenium, copper, yttrium, erbium, gallium and samarium. The utilisable alkyl groups for R' can be linear or branched, and preferably they have 1 to 5 carbon atoms .
According to the invention, the polysaccharide can be soluble, or in the form of microparticles. According to the invention, the polysaccharide can be chosen, for example, from among natural starch, cellulose or reticulated amyl pectin.
The natural starch can, for example, be maize starch.
The polysaccharide can be in the form of microparticles, for example in the form of microspheres .
The present inventors have also demonstrated that modified cellulose according to the present invention offers very good pulmonary captation and an elimination speed slower than with starch. The modified cellulose of the present invention can therefore also be used for radiotherapy, for example with labelling with rhenium, copper, or with one of the above-mentioned metals, since it corresponds to the radiotherapy necessity of using microparticles with a longer half-life.
According to the invention, the sequestering groups can be chosen for example from the groups with formulae:
Figure imgf000008_0001
in which R1, R2, R3, R4 and R5 are independently hydrogen atoms , saturated or unsaturated hydrocarbonic groups , carboxylic groups or aromatic groups,
Figure imgf000009_0001
is an aromatic nucleus possibly containing one or several hetero-atoms, and n is a whole number between 1 and 5.
For example, they can be chosen from among the formula groups :
^
N •NH- N N
SCH, SCH,
CH,
<
N NH C , =N NH C
^ NH c H NHCH3
Figure imgf000010_0001
NH NH
According to the invention, the microparticles, for example in the shape of microspheres, can be of a dimension from 0.01 to 100 μm, preferably from 10 to 50 μm for diagnosis by pulmonary scintigraphy and from 0.1 to 5 μm for therapy.
According to the invention, the levels of sequestering groups can be from 0.1 to 50% compared to the saccharide patterns of the polysaccharide, preferably from 2 to 15%.
According to the invention, in the radiopharmaceutical product, particularly when it is used for diagnosis, the radioactive metal can be 99Tc or gallium-67.
This can be the case, for example, when the radiopharmaceutical product is used for pulmonary scintigraphy.
According to the invention, in the radiopharmaceutical product, in particular when it is used for therapy, the radioactive metal can be rhenium-
186 or 188, copper-64 or 67, yttrium-90, erbium-169 or samarium-153.
According to the invention, said radiopharmaceutical product can be in the form of a suspension of microspheres in a physiologically acceptable liquid or in lyophilised form. The present invention also provides a preparation procedure for the radiopharmaceutical product of the invention comprising the following stages:
(a) submit a polysaccharide, for example such as those mentioned above, to oxidation controlled by means of a periodate,
(b) react the oxidised polysaccharide with a compound containing a primary amine function or hydrazin of formula R-NH2 or
R N NH.
R' (c) in which R is a hydrocarbonic or aromatic group comprising at least one atom of sulphur, in order to bond in a covalent manner to the polysaccharide with sequestering groups the metals of formulae R-NH- , R- N= or R-NH-N=, and R' is a hydrogen atom or an alkyl grouping, for example methyl, (d) react the polysaccharide comprising the sequestering groups with a salt of a radioactive metal chosen from among technetium, rhenium, copper, yttrium, erbium and samarium.
The oxidation controlled by means of a periodate can be that described, for example, in CL. Mehltretter, "Methods in Carbohydrate Chemistry", vol. IV, 1964, applied in particular to the oxidation of starch, dextrane or cellulose. It is used in the examples given below.
According to the invention, the compound containing a primary amine function can correspond to the formula NH2- (CH2) n-SH with n being a whole number between 1 and 5, and can include a supplementary reducing stage of this compound with sodium borohydride between stages (b) and (c) .
According to the invention, the compound bonded to the polysaccharide can correspond, for example, to one of the following formulae:
Figure imgf000013_0001
S
^ <
NH, NH C , NH NH C NHC6H5 NHCH3
NH2 NH C , NH2 NH C
^N (CH3 ) 2 ^ NH CH2CH=CH2
Figure imgf000013_0002
N C SH
NH2 C II N II
NH
According to the invention, the level of sequestering groups fixed on the polysaccharide can be regulated by controlling the level of oxidation of the polysaccharide in stage (a) mentioned above. This oxidation level of polysaccharide can, for example, be between 10 to 50%. The level of the sequestering group can, for example, be between 2 and 15%. In order to allow labelling of the polysaccharide according to the invention, for example with 99mTc, the inventors have therefore used a two-stage transformation method.
This method can be presented as consisting of carrying out a controlled oxidation of the polysaccharide by the periodate in the first stage. Each unit of oxidised glucose thus generates two aldehyde groups in neighbouring positions following the chemical reaction diagram given below:
Figure imgf000014_0001
natural glucoside monomer oxidised glucoside monomer
The level of oxidation of the polysaccharide can be variable and easily adjusted. In fact, the yield of this oxidation reaction is close to 100% and the level of oxidaton can be calculated from the quantities of periodate added. In general, oxidation levels lower than 50% are used so as to modify the structure of the macromolecule only slightly. The real oxidation level, ranging from 1 to 100%, can easily be determined by a colorometric method.
In the second stage, the oxidised polysaccharide is made to react with a molecule containing an amine or hydrazin grouping with the general formula RNH2 or RNHNH2 to form a chelating grouping able to sequester technetium. Thus one obtains a Schiff base type ligand or thiosemicarbazone .
This second stage can be summarised as follows: 0
\
R NH.
H aldehyde grouping amine or hydrazin
ϊ
R C N
H functionalised aldehyde grouping
with:
1. R=NRχ(C=S) SR2 (Schiff bases issued from dithiocarbazate
2. R=NRi(C=S)NR2R3 (thiosemicarbazones)
3. R= aromatic grouping (aromatic Schiff bases)
4. R= alkyl grouping (alkylic Schiff bases); in this case the Schiff bases are not stable and one carries out a second reduction stage of the C=N bond with borohydride so as to stabilise it and then an amine C-NHR bond is obtained. According to the invention, stage (c) can for example consist of putting into contact the microspheres of polysaccharide comprising the sequestering groups for example with a solution of pertechnetate 99mTc04 " in the presence of a reducing agent, for example stannous chloride.
According to the invention, microparticles, for example microspheres, for example maize starch or starch with a base of reticulated amyl pectin can thus be oxidised, then coupled to a molecule containing an amine or hydrazin function, for example S-methyl dithiocarbazate. These particles modified in this way can easily be labelled with, for example, 99mTc .
The present invention thus provides in particular microparticles prepared for example from a base of starch particles, which therefore do not present the inconveniences of the albumin mentioned above. In addition, the starch is described as an excipient in the pharmacopoeia. It is therefore easily available and at low cost.
The microparticles of the present invention also have the advantage of being able to be sterilised easily, for example by irradiation, and to be processed under the form of a kit ready for labelling.
Moreover, the present inventors have demonstrated according to the present invention that the speed of pulmonary clearance can be modified according to the level of oxidation of the microparticles used in the present invention, which is not possible, for example, with human albumin microspheres.
Another advantage of the present invention lies in the simplicity of operation of the procedure: the reaction conditions being very gentle: reactions at ambient temperature, in an aqueous medium, quasi- quantitative yields. In addition, the sequestering reactions, for example with technetium, are quantitative; they take place at room temperature and without final purification which makes it possible to adapt to the requirements of sterility and simplicity of preparation necessary for the utilisation of technetised kits in the hospital environment.
The present invention also provides a diagnosis kit which can be used, for example, for pulmonary scintigraphy. This kit comprises: a first flask containing a polysaccharide according to the invention, that is to say provided with sequestering groups linked to the polysaccharide by covalent bonds and chosen among the groups of formulae R-NH- , R-N= and R N N =
R' in which R is a hydrocarbonic or aromatic group comprising at least one atom of sulphur, and in which R' is a hydrogen atom or an alkyl grouping such as methyl.
According to the invention, the polysaccharide can, for example, be in the form of microparticles, for example in the shape of microspheres, the microparticles being able to be in lyophilised form or in suspension in a pharmaceutically acceptable liquid.
The kit of the present invention can furthermore comprise a second flask containing stannous chloride preferably uin lyophilised form, or also when the polysaccharide is in lyophilised form, for example in the form of microparticles, in the first flask, said first flask can besides contain lyophilised stannous chloride. The kits of the present invention are stable for at least twelve months as demonstrated in the examples given below.
The radiopharmaceutical product of the present invention therefore presents all the qualities required for a use such as radiopharmaceutical usage, for example for scintigraphy of pulmonary perfusion or for radiotherapy.
Other advantages will also appear when reading the following examples related to the present invention.
EXAMPLES Example 1
A suspension of 10 g of maize starch from the pharmacopoeia is prepared, previously sieved between 10 and 40 μm, containing about 10% water, that is 0.055 mole of glucose in 100 ml of water. One adds 0.0168 mole of sodium periodate (0.3 eq) (NaI0 ) , that is 3.6 g, dissolved in 100 ml of water. The suspension is then stirred for 18 hours at room temperature. The solution is filtered and the oxidised starch is rinsed by 5 times 20 ml of water and then 2 times 50 ml of acetone. The starch is vacuum dried and one obtains 10 g of starch oxidised at 30% (yield = 100%) .
A suspension of 10 g of starch oxidised at 30% is prepared in 60 ml of a water/ethanol mixture 2/1 by volume. Next one adds 0.1 eq (0.1x0.055x2) that is 0.011 mole of S-methyl dithiocarbazate (NH2NH (C=S) SCH3) , M=122, that is 1.34 g, dissolved in 10 ml of ethanol. The suspension is stirred for 18 hours at room temperature. The solution is next filtered and the modified starch is washed by 3 times 20 ml of ethanol and then vacuum dried. One thus obtains about 10 g of modified starch. The assay of the powder by elementary analysis gives a sulphur content of 5.4%, which corresponds to a coupling level of S-methyl dithiocarbazate (DTCZ) of 7% (7 units dithiocarbazate for 100 theoretical aldehyde functions that is 14 dithiocarbazate units for 100 glucose units) . The coupling yield is therefore 70%. One thus obtains 10 g of starch oxidised at 30% and coupled to the DTCZ at 7%. Labelling reaction with 99mTc
10 mg of starch thus modified are introduced into a flask of the penicillin type. One then adds 4 ml of physiological serum then 10 μg of SnCL2, 2H20 (20 μl of a solution at 0.5 mg/ml in HCI 0.1 N) . Then 1 ml of a solution of 99Tc0" (5mc) is added. The solution is stirred for 15 minutes and the radiochemical purity control (RCP) is carried out by filtration of 1 ml of the solution on a Millipore filter of 0.22 μm then the filter is rinsed by 2ml of physiological serum. The labelled microspheres are retained on the filter contrary to the radioactive impurities not linked to the microspheres which are found in the filtrate. The radiochemical purity corresponds to:
RCP= (activity on the filter/ otal activity) xlOO It is 98.9%. Example 2
Starch modification
One proceeds as for example 1 but one uses 0.2 eq of periodate during the oxidation reaction. One obtains a starch oxidised at 20%. One proceeds in the same way as for example 1 for the coupling reaction and one obtains a starch oxidised at 20% and coupled to the DTCZ at 7%. Labelling reaction with 99mTc
One proceeds as for example 1. The radiochemical purity (RCP) is 99%. Example 3 Starch modification
One proceeds as for example 1 but one uses 0.1 eq of periodate during the oxidation reaction. One obtains a starch oxidised at 10%. One proceeds in the same way as for example 1 for the coupling reaction and one obtains a starch oxidised at 10% and coupled to the DTCZ at 7%. Labelling reaction with 99mTc
One proceeds as for example 1. The radiochemical purity (RCP) is 98.8%. Example 4
One proceeds as for example 1 to obtain 10 g of starch oxidised at 30%. A suspension is prepared of 10 g of starch oxidised at 30% in 60 ml of a mixture of water/ethanol (2/1 by volume). Next one adds 0.1 eq (0.1x0.055x2), that is 0.011 mole of N-methyl-S-methyl dithiocarbazate (NH2N(CH3) (C=S) SCH3) , M=136, that is 1.50 g dissolved in 10 ml of ethanol. The suspension is stirred for 18 hours at room temperature. The solution is then filtered and the modified starch washed by 3 times 20 ml ethanol and then vacuum dried. One thus obtains about 10 g of modified starch. The assay of the powder by elementary analysis gives a sulphur content of 5%, which corresponds to a coupling level of the N- methyl S-methyl dithiocarbazate of 6.5% (6.5 units dithiocarbazate for 100 theoretical aldehyde functions, that is 13 dithiocarbazate units for 100 units glucose). The coupling yield is thus 65%. Labelling reaction to 99Tc
One proceeds as for example 1. The RCP is 95%. Example 5 Starch modification
One proceeds as for example 1 to obtain 10 g of starch oxidised at 30%. A suspension is prepared of
10 g of starch oxidised at 30% in 60 ml of a mixture of water/ethanol (2/1 by volume). Next one adds 0.1 eq
(0.1x0.055x2), that is 0.011 mole of 4-phenyl 3- thiosemicarbazide (NH2NH) (C=S) NH (C6H5) , M=167, that is 1.83 g dissolved in 10 ml of ethanol. The suspension is stirred for 18 hours at room temperature. The solution is then filtered and the modified starch washed by 3 times 20 ml ethanol and then vacuum dried. One thus obtains about 10 g of modified starch. The assay of the powder by elementary analysis gives a sulphur content of 3.16%, which corresponds to a coupling level of the 4-phenyl 3-thiosemicarbazone of 8% (8 units thiosemicarbazide for 100 theoretical aldehyde functions, that is 16 thiosemicarbazone units for 100 units glucose). The coupling yield is thus 80%. Labelling reaction with 99mTc One proceeds as for example 1. The RCP is 98%. Example 6
Starch modification
One proceeds as for example 1 to obtain 10 g of starch oxidised at 30%. A suspension is prepared of
10 g of starch oxidised at 30% in 60 ml of a mixture of water/ethanol (2/1 by volume). Next one adds 0.1 eq
(0.1x0.055x2), that is 0.011 mole of 4-methyl 3- thiosemicarbazide (NH2NH) (C=S)NH (CH3) , M=105, that is 1.15 g dissolved in 10 ml of ethanol. The suspension is shaken for 18 hours at room temperature. The solution is then filtered and the modified starch washed by 3 times 20 ml ethanol and then vacuum dried. One thus obtains about 10 g of modified starch. The assay of the powder by elementary analysis gives a sulphur content of 2.9%, which corresponds to a coupling level of the 4-methyl 3-thiosemicarbazone of 7.3% (7.3 units thiosemicarbazide for 100 theoretical aldehyde functions, that is 14.6 thiosemicarbazone units for 100 units glucose). The coupling yield is thus 73%. Labelling reaction with 99mTc
One proceeds as for example 1. The RCP is 97%. Example 7 Starch modification One proceeds as for example 1 to obtain 10 g of starch oxidised at 30%. A suspension is prepared of
10 g of starch oxidised at 30% in 60 ml of a mixture of water/ethanol (2/1 by volume). Next one adds 0.1 eq
(0.1x0.055x2), that is 0.011 mole of 4,4-dimethyl 3- thiosemicarbazide (NH2NH) (C=S) N (CH3) 2, M=119, that is 1.30 g dissolved in 10 ml of ethanol. The suspension is stirred for 18 hours at room temperature. The solution is then filtered and the modified starch washed by 3 times 20 ml ethanol and then vacuum dried. One thus obtains about 10 g of modified starch. The assay of the powder by elementary analysis gives a sulphur content of 3%, which corresponds to a coupling level of the 4,4-dimethyl 3-thiosemicarbazone of 7.5% (7.5 units thiosemicarbazide for 100 theoretical aldehyde functions, that is 15 thiosemicarbazone units for 100 units glucose). The coupling yield is thus 75%. Labelling reaction with 99mTc
One proceeds as for example 1. The RCP is 96%. Example 8 Starch modification One proceeds as for example 1 to obtain 10 g of starch oxidised at 30%. A suspension is prepared of
10 g of starch oxidised at 30% in 60 ml of a mixture of water/ethanol (2/1 by volume). Next one adds 0.1 eq
(0.1x0.055x2), that is 0.011 mole of 4-allyl 3- thiosemicarbazide (NH2NH) (C=S) NH (CH2CH=CH2) , M=131, that is 1.44 g dissolved in 10 ml of ethanol. The suspension is stirred for 18 hours at room temperature. The solution is then filtered and the modified starch washed by 3 times 20 ml ethanol and then vacuum dried. One thus obtains about 10 g of modified starch. The assay of the powder by elementary analysis gives a sulphur content of 3%, which corresponds to a coupling level of the 4-allyl 3-thiosemicarbazone of 7.5% (7.5 units thiosemicarbazide for 100 theoretical aldehyde functions, that is 15 thiosemicarbazone units for 100 units glucose). The coupling yield is thus 75%. Labelling reaction with 99mTc
One proceeds as for example 1. The RCP is 98%. Example 9
Starch modification One proceeds as for example 1 to obtain 10 g of starch oxidised at 30%. A suspension is prepared of 10 g of starch oxidised at 30% in 60 ml of a mixture of water/ethanol (2/1 by volume). Next one adds 0.1 eq (0.1x0.055x2), that is 0.011 mole of 3- thiosemicarbazide (NH2NH) (C=S)NH2, M=91, that is 1 g dissolved in 10 ml of ethanol. The suspension is stirred for 18 hours at room temperature. The solution is then filtered and the modified starch washed by 3 times 20 ml ethanol and then vacuum dried. One thus obtains about 10 g of modified starch. The assay of the powder by elementary analysis gives a sulphur content of 2.9%, which corresponds to a coupling level of the 3-thiosemicarbazone of 7.3% (7.3 units thiosemicarbazide for 100 theoretical aldehyde functions, that is 14.6 thiosemicarbazone units for 100 units glucose). The coupling yield is thus 73%. Labelling reaction with 99mTc
One proceeds as for example 1. The RCP is 95%. Example 10 Starch modification
One proceeds as for example 1 to obtain 10 g of starch oxidised at 30%. A suspension is prepared of 10 g of starch oxidised at 30% in 60 ml of a mixture of water/ethanol (2/1 by volume). Next one adds 0.1 eq (0.1x0.055x2), that is 0.011 mole of 2-aminothiophenol
(C6H4) (NH2) (SH) , M=125, that is 1.37 g dissolved in 10 ml of ethanol. The suspension is stirred for 18 hours at room temperature. The solution is then filtered and the modified starch washed by 3 times 20 ml ethanol and then vacuum dried. One thus obtains about 10 g of modified starch. The assay of the powder by elementary analysis gives a sulphur content of 3%, which corresponds to a coupling level of the 2- aminothiophenol of 7.5% (7.5 units aminothiophenol for 100 theoretical aldehyde functions, that is 15 aminothiophenol units for 100 units glucose) . The coupling yield is thus 75%. Labelling reaction with 99mTc
One proceeds as for example 1. The RCP is 94%. Example 11 Starch modification
One proceeds as for example 1 to obtain 10 g of starch oxidised at 30%. A suspension is prepared of 10 g of starch oxidised at 30% in 60 ml of a mixture of water/ethanol (2/1 by volume). Next one adds 0.1 eq (0.1x0.055x2), that is 0.011 mole of 2- mercaptoethylamine (or 2-aminoethanethiol)
(NH2CH2CH2SH) , M=91, that is 1 g dissolved in 10 ml of ethanol. The suspension is stirred for 18 hours at room temperature. Next one adds 0.015 mole of sodium borohydride (NaBH4) so as to reduce the Schiff base formed to stabilise it (non-aromatic Schiff bases not being stable) and leaves it to react for 1 hour. The solution is then filtered and the modified starch washed by 3 times 20 ml ethanol and then vacuum dried. One thus obtains about 10 g of modified starch. The assay of the powder by elementary analysis gives a sulphur content of 3.4%, which corresponds to a coupling level of the 2-mercaptoethylamine of 8.5% (8.5 units 2-mercaptoethylamine for 100 theoretical aldehyde functions, that is 17 2-mercaptoethylamine units for 100 units glucose). The coupling yield is thus 85%. Labelling reaction with 99mTc
One proceeds as for example 1. The RCP is 95%. Example 12 Starch modification One proceeds as for example 1 to obtain 10 g of starch oxidised at 30%. A suspension is prepared of
10 g of starch oxidised at 30% in 60 ml of a mixture of water/ethanol (2/1 by volume). Next one adds 0.1 eq
(0.1x0.055x2), that is 0.011 mole of 2-amino 4- mercaptotriazole (C4N2H) (SH) (NH2) , M=116, that is 1.27 g dissolved in 10 ml of ethanol. The suspension is stirred for 18 hours at room temperature. The solution is then filtered and the modified starch washed by 3 times 20 ml ethanol and then vacuum dried. One thus obtains about 10 g of modified starch. The assay of the powder by elementary analysis gives a sulphur content of 2.8%, which corresponds to a coupling level of the 2-amino 4-mercaptotriazole of 7% (7 units mercaptotriazole for 100 theoretical aldehyde functions, that is 14 mercaptotriazole units for 100 units glucose) . The coupling yield is thus 75%. Labelling reaction with 99mTc
One proceeds as for example 1. The RCP is 85%. Comparative example 1 One uses 10 g of sieved pharmacopoeia maize which has not undergone chemical transformation and one caries out the same procedure as in example 1 to label it with 99mTc.
The RCP is 19%.
This example is a good demonstration of the fact that the chemical modification (fixation of sequestering groups) carried out according to the invention is certainly necessary to allow labelling with 99mTc. In addition, one cannot obtain durable pulmonary captation if the microparticles are labelled without prior chemical transformation contrary to the product relating to the present invention. These results are therefore in contradiction with that which is described in FR-A-2 273 516. Example 13 Cellulose modification
One proceeds as in example 1 but using sieved cellulose between 10 and 40 μm. Thus one obtains 10 g of cellulose oxidised at 30% and coupled to the DTCZ at 7%. Labelling reaction to 99mTc
One proceeds as in example 1. The RCP is 99.1%. Example 14
Sprague Dawley rats weighing about 200 g are anaesthetised with sodium thiopental, and are injected intravenously with different solutions of microspheres labelled with 99Tc according to examples 1 to 9 and 13. Each animal receives 0.2 ml of solution in the penis vein, that is 0.2 mc per animal. The animals are then placed under a gamma-ray camera and successive static images are shot over a period of 3 hours. One thus obtains successive images after acquiring 15,000 shots per image. Then, manually, one defines the zones of interest in order to estimate the activity present in the different organs 15 minutes after the injection. The results are given in tables I below.
Tables I : Results
Figure imgf000028_0001
Figure imgf000028_0002
One thus notes that the modified microspheres show very good pulmonary captation. In addition, one can modulate the speed of pulmonary elimination by varying the oxidation level as shown in examples 1 , 2 and 3 (oxidation levels 30, 20 and 10%).
The usage of cellulose makes it possible to lengthen the speed of elimination considerably (example 10, half-life > 4 hours) . Comparative example 2
In this example, natural starch is not used, but microspheres prepared from amyl pectin reticulated by epichlorhydrin as in the patent FR-A-2 273 516. Preparation of reticulated microspheres of starch
One dissolves 8 g of maize amyl pectin in 40 ml of a solution containing 4 g of NaOH and 0.15 g of sodium borohydride. The amyl pectin is left for 24 hours to dissolve. Next one prepares an emulsion by stirring 60 ml of fluid paraffin and 1.6 g of soy lecithin dissolved in 4 ml of hexane at 800 revs/min. Then one adds the aqueous phase containing the amyl pectin and then 3.2 ml of epichlorhydrin. The emulsion is heated to 55°C for 4 hours and then left to be stirred overnight. The microspheres obtained of a size around 50 μm are washed by 3 times 250 ml of acetone, dried and then lyophilised. Labelling with 99mTc
One proceeds as for example 1 but using 1 mg of SnCl2, 2H20. The RCP is 90%. Starch modification
One proceeds as for example 1 but using 10 g of microspheres of amyl pectin reticulated by the epichlorhydrin previously prepared. One thus obtains 10 g of microspheres of amyl pectin oxidised at 30% and coupled to the DTCZ at 7%. Labelling reaction with 99Tc
One proceeds as for example 1. The RCP is 99%. Example 15
One follows the same operational mode as in example 14 to test the microspheres of reticulated amyl pectin labelled with 99mTc of the comparative example 2. The results obtained are given in table II below.
Table II
Figure imgf000030_0001
One notes that contrary to the description in FR- A-2 273 516 the microspheres of reticulated amyl pectin not modified chemically are labelled by 99mTc but do not present any pulmonary captation, doubtless due to the weak link between 99Tc and the microspheres. On the other hand, these microspheres transformed chemically by the procedure of the invention demonstrate good pulmonary captation. Example 16
Starch microspheres prepared as in example 1 (starch oxidised at 30%, coupled with DTCZ at 7%) are used to produce sterile labelling kits and are ready for labelling with 99mTc . Sterilisation of the microspheres
10 g of microspheres are introduced into a flask crimped and then irradiated by a source of cobalt-60. The microspheres receive a total dose of gamma radiation of 25kGy over 20 hours. Preparation of the kits
200 mg of sterilised microspheres are introduced in a sterile way into a reactor containing 20 ml of NaCl 9/1000. The solution is de-aerated by nitrogen bubbling and then one adds 400 μl of a sterile solution of SnCl2, 2H20 at 0.5 mg/ml in HCI 0.1N. One separates 1 ml of solution for each of 20 flasks, which are then lyophilised and placed in a nitrogen atmosphere.
Each flask thus contains :
10 mg of modified microspheres 10 μg of SnCl2, 2H20 9 mg of NaCl Labelling reaction with 99mTc
5 ml of Tc0 " (5mc) solution is added to a flask in lyophilised form and left to react for 15 minutes.
One proceeds as in example 1. The RCP is 98.7%. Kit stability trial
Labelling kits prepared as above are stored at different temperatures and then the labelling reaction to 99mTc is tested so as to evaluate their stability. The results obtained are given in table III below:
Table III : RCP (%)
Figure imgf000031_0001
One notes the very high stability of the kit stored at a temperature between 2 and 8°C. Example 17
Sprague Dawley rats weighing about 200 g are anaesthetised with sodium thiopental, and are then injected intravenously with different solutions of microspheres labelled with 99mTc as in example 14. Each animal receives 0.2 ml of solution in the penis vein, that is 0.2 mc per animal. The animals are then killed 15 minutes after the injection. Next, the different organs are retrieved, the measurement of radioactivity present in each organ is counted and thus the percentage of activity present in each organ is determined. The results are given in table IV below:
Table IV - Results of the dose injected 15 minutes after injection
Figure imgf000032_0001
One thus notes a very high pulmonary captation whereas as there is weak captation in the other organs for natural starch microspheres as well as reticulated starch base microspheres. The sterile product prepared under kit form thus seems completely adapted to usage as a radiopharmaceutical product for pulmonary perfusion substituting for albumin, and for radiotherapy. Example 18
One uses cellulose oxidised at 30% and coupled with the DTCZ at 7% as in example 13. The cellulose thus modified is labelled with rhenium 186 in order to illustrate the utilisation of the support according to the present invention for therapy. Labelling reaction with Re 186
10 mg of modified cellulose are introduced into a penicillin type flask. One then adds 2 ml of physiological serum and then 20 mg of citric acid and finally 1 mg of SnCl2, 2H20 (100 μl of a solution at
10 mg/ml in HCI 0.1 N) .
Next one adds to the contents of the flask 0.1 ml of Re04 " solution corresponding to an activity of 2 mc . The flask is heated in a water bath at 100°C for 30 minutes. The radiochemical purity (RCP) is determined by filtration on a Millipore as in example 1. RCP = 92%
In order to prove the stability of the link between rhenium 186 and the cellulose microspheres, a stability test in vi tro is carried out.
The mixture is incubated with HSA (human serum albumin) at 20 mg/ml at 37°C. The following results are obtained:
Figure imgf000033_0001
These results therefore prove the very high stability of labelling of cellulose microspheres with rhenium 186 and the high stability of the microspheres themselves vis-a-vis HSA. The expert will easily understand that these results can be extrapolated to the utilisation of rhenium 188. Example 19
Sprague Dawley rats weighing about 200 g are anaesthetised and then injected with 0.2 ml (0.1 mc) of cellulose microsphere solution labelled with Re 186 as in example 18.
The animals are then placed under a gamma-ray camera and images are registered over 48 hours.
The activity present in the zones of interest is then calculated as in example 14.
Figure imgf000034_0001
These results therefore prove that the activity remains blocked at the pulmonary level for at least 48 hours. This type of microsphere can thus be used for therapy.
The most important clinical application can be the treatment of liver cancers after injection, not intravenously but directly into the hepatic artery (metabolic radiotherapy) .
Another possible application is to inject this type of particle subcutaneously in breast cancer. The particles migrating through the lymphatic system may make it possible to treat the sentinel nodes invaded by cancerous cells .

Claims

1. A radiopharmaceutical product comprising a polysaccharide provided with sequestering groups linked to the polysaccharide by covalent bonds and chosen from among the groups of formulae R-NH-, R-N=, and R N N
R' in which R is a hydrocarbonic or aromatic group comprising at least one atom of sulphur, and R' is an atom of hydrogen or an alkyl or methyl grouping, said sequestering groups forming, together with a radioactive metal chosen from among technetium, rhenium, copper, yttrium, erbium, gallium and samarium, a complex of the chelate type, in which the polysaccharide is in the form of microparticles.
2. A radiopharmaceutical product according to Claim 1 in which the sequestering groups are chosen from among the groups of formulae:
Figure imgf000036_0001
in which R1, R2, R3 , R4 and R5 are independently atoms of hydrogen atoms, saturated or unsaturated hydrocarbonic groups, carboxylic groups or aromatic groups,
Figure imgf000037_0001
is an aromatic nucleus possibly containing one or several heteroatoms, and n is a whole number between
1 and 5.
3. A radiopharmaceutical product according to Claim 2 in which the sequestering groups are chosen from among the groups of formulae:
Figure imgf000037_0002
Figure imgf000038_0001
N SH N SH
■NH' and N
\ / \ /
NH NH
4. A radiopharmaceutical product according to any one of Claims 1 to 3 , in which the polysaccharide is chosen from among natural starch, cellulose and reticulated amyl pectin.
5. A radiopharmaceutical product according to any one of Claims 1 to 5 , in which the microparticles have a dimension between 0.01 and 100 urn
6. A radiopharmaceutical product according to any one of Claims 1 to 5 , in which the level of sequestering groups is from 0.1 to 50% relative to the saccharide patterns of polysaccharide.
7. Utilisation of a radiopharmaceutical product according to any one of Claims 1 to 6, in which the radioactive metal is 99mTc or 67Ga to prepare a product intended for diagnosis.
8. Utilisation of a radiopharmaceutical product according to any one of Claims 1 to 6, in which the radioactive metal is rhenium-186 or 188, copper-64 or 67, yttrium 90, erbium 169 or samarium 153, to prepare a drug.
9. Utilisation of a radiopharmaceutical product according to any one of Claims 1 to 7, in which the radioactive metal is 99Tc to prepare a product intended for pulmonary scintigraphy.
10. A radiopharmaceutical product according to any one of Claims 1 to 6 , under the form of a suspension of microspheres in a physiologically acceptable liquid or in lyophilised form.
11. A procedure for preparation of a radiopharmaceutical product according to any one of Claims 1 to 6, which comprises the following stages:
(a) submit a polysaccharide to an oxidation carried out by means of a periodate, (b) make the oxidated polysaccharide react with a compound containing a primary amine function or hydrazin of formula R-NH2 or R N H,
R'
in which R is a hydrocarbonic or aromatic group comprising at least one atom of sulphur, in order to bond in a covalent manner to the polysaccharide with sequestering groups the metals of formulae R-NH- , R-N= or R-NH-N=, and R' is a hydrogen atom or an alkyl or methyl grouping .
(c) make the polysaccharide comprising the sequestering groups react with a salt of a radioactive metal chosen from among technetium, rhenium, copper, yttrium, erbium and samarium.
12. A procedure according to Claim 11, in which the compound containing a primary amine function corresponds to the formula NH2- (CH2) n-SH with n being a whole number from 1 to 5, and comprising a supplementary stage of reduction of this compound by sodium borohydride between stages (b) and (c) .
13. A procedure according to Claim 11, in which the compound bonded to the oxidised polysaccharide corresponds to one of the following formulae: NH2 — NH - - c M NH, N — -c M
^ SCH3 2 1 ^ SCH3
CH3
NH2 - — NH — - c M Nlfcj — NH — -c M
^ NHCcHq ^ NHCH3
NH2 - — NH — -
Figure imgf000041_0001
Figure imgf000041_0002
N c SH
NH2 c N
\ /
NH
14. A procedure according to any one of Claims 11 to 13 , in which the level of sequestering groups fixed on the polysaccharide is regulated by controlling the level of oxidation of the polysaccharide in stage (a) .
15. A procedure according to Claim 14, in which the oxidation level of the polysaccharide is from 10 to
50%.
16. A procedure according to Claim 14, in which the level of sequestering groups is from 2 to 15%.
17. A procedure according to any one of Claims 10 to 16, in which the stage (c) consists of putting into contact microspheres of polysaccharide comprising sequestering groups with a solution of pertechnetate 99ιnTc0", in the presence of a reducing agent.
18. A diagnosis kit which can be used for pulmonary scintigraphy which comprises: a first flask containing a polysaccharide provided with sequestering groups linked to said polysaccharide by covalent bonds and chosen among the formulae groups R-NH-, R-N= and
R N N =
R' in which R is a hydrocarbonic or aromatic group comprising at least one atom of sulphur, and in which R' is an atom of hydrogen or an alkyl or methyl grouping, in which the polysaccharide is in the form of lyophilised microparticles or in suspension in a pharmaceutically acceptable liquid.
19. A kit according to Claim 18 comprising also a second flask containing stannous chloride in lyophilised form.
20. A kit according to Claim 18, in which the polysaccharide being in the form of lyophilised microparticles in the first flask, said first flask also contains lyophilised stannous chloride.
PCT/IB2000/001161 1999-09-01 2000-08-23 Radiopharmaceutical products and their preparation procedure WO2001015746A1 (en)

Priority Applications (17)

Application Number Priority Date Filing Date Title
AU64631/00A AU6463100A (en) 1999-09-01 2000-08-23 Radiopharmaceutical products and their preparation procedure
MXPA02001923A MXPA02001923A (en) 1999-09-01 2000-08-23 Radiopharmaceutical products and their preparation procedure.
PL00353803A PL353803A1 (en) 1999-09-01 2000-08-23 Radiopharmaceutical products and their preparation procedure
CA002383517A CA2383517A1 (en) 1999-09-01 2000-08-23 Radiopharmaceutical products and their preparation procedure
EA200200307A EA200200307A1 (en) 1999-09-01 2000-08-23 RADIOACTIVE PHARMACEUTICAL PRODUCTS AND METHOD FOR THEIR PRODUCTION
EP00951784A EP1210127A1 (en) 1999-09-01 2000-08-23 Radiopharmaceutical products and their preparation procedure
IL14807600A IL148076A0 (en) 1999-09-01 2000-08-23 Radiopharmaceutical products including a polysaccharide
JP2001520157A JP2003508455A (en) 1999-09-01 2000-08-23 Radiopharmaceutical products and methods for their preparation
HU0202714A HUP0202714A2 (en) 1999-09-01 2000-08-23 Radiopharmaceutical products and their preparation procedure
KR1020027002820A KR20020040799A (en) 1999-09-01 2000-08-23 Radiopharmaceutical Products and Their Preparation Procedure
SK277-2002A SK2772002A3 (en) 1999-09-01 2000-08-23 Radiopharmaceutical products and their preparation procedure
BR0013729-4A BR0013729A (en) 1999-09-01 2000-08-23 Radiopharmaceuticals and their preparation procedures
NZ517377A NZ517377A (en) 1999-09-01 2000-08-23 Radiopharmaceutical products and their preparation procedure
EEP200200105A EE200200105A (en) 1999-09-01 2000-08-23 Radiopharmaceuticals and Method of Preparation
BG06438A BG106438A (en) 1999-09-01 2002-02-26 Radiopharmaceutical products and method for their preparation
NO20021001A NO20021001L (en) 1999-09-01 2002-02-28 Radiopharmaceutical products and processes for their preparation
HK02106423.2A HK1044893A1 (en) 1999-09-01 2002-08-30 Radiopharmaceutical products and their preparation procedure

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
FR99/10970 1999-09-01
FR9910970A FR2797769B1 (en) 1999-09-01 1999-09-01 RADIOPHARMACEUTICAL PRODUCTS AND THEIR PREPARATION PROCESS

Publications (1)

Publication Number Publication Date
WO2001015746A1 true WO2001015746A1 (en) 2001-03-08

Family

ID=9549465

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/IB2000/001161 WO2001015746A1 (en) 1999-09-01 2000-08-23 Radiopharmaceutical products and their preparation procedure

Country Status (23)

Country Link
EP (1) EP1210127A1 (en)
JP (1) JP2003508455A (en)
KR (1) KR20020040799A (en)
CN (1) CN1371292A (en)
AU (1) AU6463100A (en)
BG (1) BG106438A (en)
BR (1) BR0013729A (en)
CA (1) CA2383517A1 (en)
CZ (1) CZ2002782A3 (en)
EA (1) EA200200307A1 (en)
EE (1) EE200200105A (en)
FR (1) FR2797769B1 (en)
HK (1) HK1044893A1 (en)
HU (1) HUP0202714A2 (en)
IL (1) IL148076A0 (en)
MX (1) MXPA02001923A (en)
NO (1) NO20021001L (en)
NZ (1) NZ517377A (en)
PL (1) PL353803A1 (en)
SK (1) SK2772002A3 (en)
WO (1) WO2001015746A1 (en)
YU (1) YU14202A (en)
ZA (1) ZA200201057B (en)

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102977174A (en) * 2012-12-19 2013-03-20 北京师范大学 99mTc(CO)3 nuclear-labeled macrocyclic polyamine triazole ring glucose-based complex as well as preparation and application of complex
EP3711781A1 (en) 2019-03-19 2020-09-23 Nanothea Spolka Akcyjna Method for preparation of radioisotope chelating polymer nanoparticles for use in diagnostics and treatment
US11793912B2 (en) 2004-09-28 2023-10-24 Atrium Medical Corporation Cross-linked fatty acid-based biomaterials

Families Citing this family (15)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7462366B2 (en) 2002-03-29 2008-12-09 Boston Scientific Scimed, Inc. Drug delivery particle
US7842377B2 (en) 2003-08-08 2010-11-30 Boston Scientific Scimed, Inc. Porous polymeric particle comprising polyvinyl alcohol and having interior to surface porosity-gradient
US8012454B2 (en) 2002-08-30 2011-09-06 Boston Scientific Scimed, Inc. Embolization
US7883490B2 (en) 2002-10-23 2011-02-08 Boston Scientific Scimed, Inc. Mixing and delivery of therapeutic compositions
US7976823B2 (en) 2003-08-29 2011-07-12 Boston Scientific Scimed, Inc. Ferromagnetic particles and methods
US7901770B2 (en) 2003-11-04 2011-03-08 Boston Scientific Scimed, Inc. Embolic compositions
US7736671B2 (en) 2004-03-02 2010-06-15 Boston Scientific Scimed, Inc. Embolization
US8173176B2 (en) 2004-03-30 2012-05-08 Boston Scientific Scimed, Inc. Embolization
US7311861B2 (en) 2004-06-01 2007-12-25 Boston Scientific Scimed, Inc. Embolization
US7727555B2 (en) 2005-03-02 2010-06-01 Boston Scientific Scimed, Inc. Particles
US7858183B2 (en) 2005-03-02 2010-12-28 Boston Scientific Scimed, Inc. Particles
US7963287B2 (en) 2005-04-28 2011-06-21 Boston Scientific Scimed, Inc. Tissue-treatment methods
US9463426B2 (en) 2005-06-24 2016-10-11 Boston Scientific Scimed, Inc. Methods and systems for coating particles
US7947368B2 (en) 2005-12-21 2011-05-24 Boston Scientific Scimed, Inc. Block copolymer particles
FR2919189A1 (en) * 2007-07-26 2009-01-30 Cyclopharma Sa Lab NOVEL COMPOSITIONS BASED ON POLYOSIDES GRAFTED BY POLYAMINE OR POLYSOUFRE COMPOUNDS.

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
FR2285857A1 (en) * 1974-09-27 1976-04-23 Pharmacia Fine Chemicals Ab MEDICAL CONTROL AGENT INTENDED TO BE ADMINISTERED BY INTRAVASCULAR ROUTE
WO1994005203A1 (en) * 1992-09-04 1994-03-17 The General Hospital Corporation Biocompatible polymers containing diagnostic or therapeutic moieties
WO1995014492A2 (en) * 1993-11-29 1995-06-01 Access Pharmaceuticals, Inc. In vivo agents comprising radioactive metal-ion chelates with acidic saccharides and glycosaminoglycans, giving improved site-selective localization, uptake mechanism, sensitivity and kinetic-spatial profiles
WO1996000588A1 (en) * 1994-06-28 1996-01-11 Nycomed Imaging A/S Low viscosity chelating polymers
FR2736834A1 (en) * 1995-07-17 1997-01-24 Cis Bio Int RADIOPHARMACEUTICAL PRODUCTS WITH TROPISMAL HEART CONTAINING A NITRURO COMPLEX OF A TRANSITION METAL AND HAVE A RAPID MYOCARDIC CLEARANCE

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
FR2285857A1 (en) * 1974-09-27 1976-04-23 Pharmacia Fine Chemicals Ab MEDICAL CONTROL AGENT INTENDED TO BE ADMINISTERED BY INTRAVASCULAR ROUTE
WO1994005203A1 (en) * 1992-09-04 1994-03-17 The General Hospital Corporation Biocompatible polymers containing diagnostic or therapeutic moieties
WO1995014492A2 (en) * 1993-11-29 1995-06-01 Access Pharmaceuticals, Inc. In vivo agents comprising radioactive metal-ion chelates with acidic saccharides and glycosaminoglycans, giving improved site-selective localization, uptake mechanism, sensitivity and kinetic-spatial profiles
WO1996000588A1 (en) * 1994-06-28 1996-01-11 Nycomed Imaging A/S Low viscosity chelating polymers
FR2736834A1 (en) * 1995-07-17 1997-01-24 Cis Bio Int RADIOPHARMACEUTICAL PRODUCTS WITH TROPISMAL HEART CONTAINING A NITRURO COMPLEX OF A TRANSITION METAL AND HAVE A RAPID MYOCARDIC CLEARANCE

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US11793912B2 (en) 2004-09-28 2023-10-24 Atrium Medical Corporation Cross-linked fatty acid-based biomaterials
CN102977174A (en) * 2012-12-19 2013-03-20 北京师范大学 99mTc(CO)3 nuclear-labeled macrocyclic polyamine triazole ring glucose-based complex as well as preparation and application of complex
CN102977174B (en) * 2012-12-19 2015-04-22 北京师范大学 99mTc(CO)3 nuclear-labeled macrocyclic polyamine triazole ring glucose-based complex as well as preparation and application of complex
EP3711781A1 (en) 2019-03-19 2020-09-23 Nanothea Spolka Akcyjna Method for preparation of radioisotope chelating polymer nanoparticles for use in diagnostics and treatment

Also Published As

Publication number Publication date
BR0013729A (en) 2002-05-07
EE200200105A (en) 2003-04-15
CZ2002782A3 (en) 2002-08-14
KR20020040799A (en) 2002-05-30
NZ517377A (en) 2003-08-29
ZA200201057B (en) 2003-07-30
MXPA02001923A (en) 2003-07-21
HUP0202714A2 (en) 2002-12-28
IL148076A0 (en) 2002-09-12
CN1371292A (en) 2002-09-25
PL353803A1 (en) 2003-12-01
CA2383517A1 (en) 2001-03-08
JP2003508455A (en) 2003-03-04
FR2797769B1 (en) 2003-07-25
HK1044893A1 (en) 2002-11-08
EA200200307A1 (en) 2002-08-29
FR2797769A1 (en) 2001-03-02
AU6463100A (en) 2001-03-26
NO20021001L (en) 2002-04-11
YU14202A (en) 2004-09-03
SK2772002A3 (en) 2002-09-10
EP1210127A1 (en) 2002-06-05
NO20021001D0 (en) 2002-02-28
BG106438A (en) 2002-09-30

Similar Documents

Publication Publication Date Title
EP1210127A1 (en) Radiopharmaceutical products and their preparation procedure
JP2894879B2 (en) Diagnostic contrast agent
EP0643706A1 (en) Compounds, compositions and methods for binding bio-affecting substances to surface membranes of bio-particles
AU691806B2 (en) Chelating agents of the type XN1S1O1 for radioactive isotopes, metal complexes thereof, and their use in diagnosis and therapy
EP2175888B1 (en) Radiopharmaceutical compositions comprising polysaccharides grafted by polyamines
US4318898A (en) Technetium-99m-labelled (2,4,5-trimethylacetanilido)-iminodiacetate for liver function diagnosis, chloroacetric acid (2,4,5-trimethylanilide), (2,4,5-trimethylacetanilido)-iminodiacetate, and process for their preparation
EP1268464A1 (en) Radioisotope-labeled complexes of glucose derivatives and kits for the preparation thereof
EP1154803B1 (en) Immobilized labeling compounds and methods
JP2564459B2 (en) Carrier for radiopharmaceutical preparation
LI et al. RADIOPHARMAZEUTISCHE ZUSAMMENSETZUNGEN DIE POLYAMINE GEPFROPFTE POLYSACCHARIDE ENTHALTEN COMPOSITIONS RADIOPHARMACEUTIQUES COMPRENANT DES POLYSACCHARIDES GREFFÉS AVEC DES POLYAMINES
JPS5842846B2 (en) Technetium-99m labeled radiological diagnostic agent for liver and bone marrow scanning and its manufacturing method
JPH0615478B2 (en) Radiopharmaceuticals and polymeric compounds for their preparation
JPS63170389A (en) Radioactive medicine and high-molecular compound for preparation thereof
CZ277786B6 (en) PROCESS FOR PREPARING RADIO DIAGNOSTIC MEANS WITH A COMPLEX 99m Tc-d,1-4,8-DIAZA-3,6,6,9-TETRAMETHYLUNDECANE-2,10-BISOXIME

Legal Events

Date Code Title Description
WWE Wipo information: entry into national phase

Ref document number: P-142/02

Country of ref document: YU

AK Designated states

Kind code of ref document: A1

Designated state(s): AE AG AL AM AT AU AZ BA BB BG BR BY BZ CA CH CN CR CU CZ DE DK DM DZ EE ES FI GB GD GE GH GM HR HU ID IL IN IS JP KE KG KP KR KZ LC LK LR LS LT LU LV MA MD MG MK MN MW MX MZ NO NZ PL PT RO RU SD SE SG SI SK SL TJ TM TR TT TZ UA UG US UZ VN YU ZA ZW

AL Designated countries for regional patents

Kind code of ref document: A1

Designated state(s): GH GM KE LS MW MZ SD SL SZ TZ UG ZW AM AZ BY KG KZ MD RU TJ TM AT BE CH CY DE DK ES FI FR GB GR IE IT LU MC NL PT SE BF BJ CF CG CI CM GA GN GW ML MR NE SN TD TG

121 Ep: the epo has been informed by wipo that ep was designated in this application
DFPE Request for preliminary examination filed prior to expiration of 19th month from priority date (pct application filed before 20040101)
WWE Wipo information: entry into national phase

Ref document number: IN/PCT/2002/00162/MU

Country of ref document: IN

WWE Wipo information: entry into national phase

Ref document number: 2000951784

Country of ref document: EP

WWE Wipo information: entry into national phase

Ref document number: 148076

Country of ref document: IL

WWE Wipo information: entry into national phase

Ref document number: 517377

Country of ref document: NZ

WWE Wipo information: entry into national phase

Ref document number: 2772002

Country of ref document: SK

Ref document number: PA/a/2002/001923

Country of ref document: MX

ENP Entry into the national phase

Ref document number: 2000 106438

Country of ref document: BG

Kind code of ref document: A

ENP Entry into the national phase

Ref document number: 2001 520157

Country of ref document: JP

Kind code of ref document: A

WWE Wipo information: entry into national phase

Ref document number: 2383517

Country of ref document: CA

Ref document number: 1020027002820

Country of ref document: KR

Ref document number: 64631/00

Country of ref document: AU

Ref document number: 008123004

Country of ref document: CN

WWE Wipo information: entry into national phase

Ref document number: PV2002-782

Country of ref document: CZ

WWE Wipo information: entry into national phase

Ref document number: P20020268A

Country of ref document: HR

WWE Wipo information: entry into national phase

Ref document number: 200200307

Country of ref document: EA

WWP Wipo information: published in national office

Ref document number: 1020027002820

Country of ref document: KR

WWP Wipo information: published in national office

Ref document number: 2000951784

Country of ref document: EP

REG Reference to national code

Ref country code: DE

Ref legal event code: 8642

WWP Wipo information: published in national office

Ref document number: PV2002-782

Country of ref document: CZ

WWP Wipo information: published in national office

Ref document number: 517377

Country of ref document: NZ

WWG Wipo information: grant in national office

Ref document number: 517377

Country of ref document: NZ

WWW Wipo information: withdrawn in national office

Ref document number: 2000951784

Country of ref document: EP