WO2003086476A1 - Derives de rotenone a marquage au technetium, et procedes d'utilisation correspondants - Google Patents

Derives de rotenone a marquage au technetium, et procedes d'utilisation correspondants Download PDF

Info

Publication number
WO2003086476A1
WO2003086476A1 PCT/US2003/008883 US0308883W WO03086476A1 WO 2003086476 A1 WO2003086476 A1 WO 2003086476A1 US 0308883 W US0308883 W US 0308883W WO 03086476 A1 WO03086476 A1 WO 03086476A1
Authority
WO
WIPO (PCT)
Prior art keywords
complex
pyridylmethyl
group
chelators
technetium
Prior art date
Application number
PCT/US2003/008883
Other languages
English (en)
Inventor
John W. Babich
Kevin P. Maresca
Original Assignee
Biostream, Inc.
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Biostream, Inc. filed Critical Biostream, Inc.
Priority to AU2003224747A priority Critical patent/AU2003224747A1/en
Publication of WO2003086476A1 publication Critical patent/WO2003086476A1/fr

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
    • A61K51/02—Preparations containing radioactive substances for use in therapy or testing in vivo characterised by the carrier, i.e. characterised by the agent or material covalently linked or complexing the radioactive nucleus
    • A61K51/04—Organic compounds
    • A61K51/08—Peptides, e.g. proteins, carriers being peptides, polyamino acids, proteins
    • A61K51/088—Peptides, e.g. proteins, carriers being peptides, polyamino acids, proteins conjugates with carriers being peptides, polyamino acids or proteins
    • 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
    • A61K51/041—Heterocyclic compounds
    • A61K51/0412—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
    • A61K51/0421—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom

Definitions

  • Coronary heart disease is the leading cause of death in the United States, accounting for roughly 24% of all deaths.
  • the cost of cardiovascular diseases in 1999 is estimated by the American Heart Association (AHA) at $286.5 billion.
  • Myocardial perfusion scintigraphy is widely used in the evaluation of patients with known or suspected coronary artery disease (CAD).
  • CAD coronary artery disease
  • the extensive clinical use of stress myocardial perfusion imaging has resulted largely from its demonstrated improved diagnostic sensitivity and specificity for detection of CAD as compared with exercise electrocardiogram.
  • myocardial flow tracers with improved tracer kinetics.
  • Zaret B and Beller GA Wintergreen Panel Summaries J. Nuclear Cardiology (1999) 6:111.
  • tracers Although several tracers are currently available for perfusion imaging, all of these tracers suffer from one or more limitations which render them less than ideal agents for assessment of cardiac perfusion (i.e., limited extraction at high flow (Tc99m-sestamibi, Tl-201 Chloride) (Marshall, R.C., Leidholdt E.M. Jr., Zhang, D. Y spirit Barnett, CA.
  • radiopharmaceuticals may be used as diagnostic or therapeutic agents by virtue of the physical properties of their constituent radionuclides. Thus, their utility is not based on any pharmacologic action.
  • Most clinically used drugs of this class are diagnostic agents incorporating a gamma-emitting nuclide which, because of physical or metabolic properties of its coordinated ligands, localizes in a specific organ after intravenous injection.
  • the resultant images can reflect organ structure or function. These images are obtained by means of a gamma camera that detects the distribution of ionising radiation emitted by the radioactive molecules.
  • the radiolabel is a gamma-radiation emitting radionuclide and the radiotracer is located using a gamma-radiation detecting camera (this process is often referred to as gamma scintigraphy).
  • the imaged site is detectable because the radiotracer is chosen either to localize at a pathological site (termed positive contrast) or, alternatively, the radiotracer is chosen specifically not to localize at such pathological sites (termed negative contrast).
  • radiopharmaceuticals which provide diagnostic images of blood flow (perfusion) in the major organs and in tumors.
  • the regional uptake of these radiopharmaceuticals within the organ of interest is proportional to flow; high flow regions will display the highest concentration of radiopharmaceutical, while regions of little or no flow have relatively low concentrations. Diagnostic images showing these regional differences are useful in identifying areas of poor perfusion, but do not provide metabolic information of the state of the tissue within the region of apparently low perfusion.
  • radionuclides are less than ideal for routine clinical use.
  • the positron-emitting isotapes such as 18 F
  • the costs of procedures based on positron- emitting isotopes are very high, and there are very few of these centers worldwide.
  • 123 I-radiopharmaceuticals may be used with widely-available gamma camera imaging systems, 123 I has a 13 hour half-life (which restricts the distribution of radiopharmaceuticals based on this isotope) and is expensive to produce. Nitroimidazoles labeled with 3 H are not suitable for in vivo clinical imaging and can be used for basic research studies only.
  • Radionuclide that emits gamma energy in the 100 to 200 keV range is preferred.
  • the physical half-life ofthe radionuclide should be as short as the imaging procedure will allow.
  • radionuclides are known to be useful for radioimaging, including Ga- 67, Tc-99m, In-Ill, 1-123, 1-125, Yb-169 and Re-186.
  • the preferred radioisotope for medical imaging is Tc-99m. Its 140 keV gamma-photon is ideal for use with widely- available gamma cameras. It has a short (6 hour) half life, which is desirable when considering patient dosimetry.
  • Tc-99m is readily available at relatively low cost through commercially-produced "Mo/Tc-99m generator systems. As a result, over 80% of all radionuclide imaging studies conducted worldwide utilize Tc-99m. See generally Reedijk J. "Medicinal Applications of heavy-metal compounds" Curr.
  • Tc(I) organometallic Tc(I) carbonyl chemistry.
  • the chemistry of [ 99 ' n Tc(OH 2 ) 3 (CO) 3 ] + has been elucidated and simplified to the point where the methods are routine and offer a practical alternative to the currently employed Tc(V) chemistry.
  • the Tc(I) method offers an attractive labeling alternative.
  • the Tc(I)(CO) 3 + core limits the number of possible coordination geometries available for Tc due to the presence of the three carbonyl groups. The facial arrangement of carbonyl ligands around the metal center also impose steric constraints on the binding possibilities ofthe remaining three sites.
  • the [ 99m Tc(OH 2 ) 3 (CO) 3 ] + complex can be readily prepared in saline under 1 atm of carbon monoxide (CO).
  • This water and air stable Tc(I) complex is a practical precursor to highly inert Tc(I) complexes, due in part to the d 6 electron configuration of the metal center.
  • the preparation ofthe organometallic tris(aquo) ion is simple and straightforward, allowing for convenient manipulation and product formation. Substitution of the labile H 2 O ligands has been shown to leave the Tc(CO) 3 + core intact.
  • This stable core has the additional advantage of being smaller and less polar than the routinely employed Tc(V)-oxo systems. This characteristic could be advantageous in biologically relevant systems where the addition of the metal center effects the size, shape, and potentially the bioactivity ofthe compounds.
  • chelators are currently employed in the binding of tecmetium, all of these tracers suffer from one or more disadvantages which render them less than ideal: HYNIC requires coligands; MAG3 may be only used with the Tc(V)-oxo species; EDTA DTPA is used primarily primarily with Tc(V)-oxo and its ability to retain label is poor. Hence, additional Technetium-99m chelators are needed. Novel radiolabeled chelators that display rapid, efficient labeling and demonstrate superior labeling retention for both Tc(N)-oxo and Tc(I)-tricarbonyl cores without the use of coligands are attractive candidates for clinical evaluation as potential chelators for biologically relevant molecules.
  • Rotenone has a high affinity for mitochondria.
  • the myocardium is an organ rich in mitochrondria. Novel technetium radiolabeled rotenone analogs that display efficient myocardial uptake and adequate myocardial retention are attractive candidates for clmical evaluation of myocardial blood flow.
  • Rotenone is a specific, high-affinity inhibitor of complex I (NADHubiquinone oxidoreductase), the proximal enzyme of the mitochondrial electron transport chain. Since rotenone inhibition defines the activity of complex I, defects in radiotracer binding can be expected to reflect functional changes in the enzyme, and hence, abnormalities ofthe mitochondrial energy metabolism.
  • X represents independently for each occurrence O or S;
  • Z represents a chelator comprising a radionuclide;
  • R represents independently for each occurrence H, lower alkyl, or halogen
  • R' represents independently for each occurrence lower alkyl
  • R" represents independently for each occurrence H, or lower alkyl
  • R 3 represents independently for each occurrence H, or lower alkyl
  • the stereochemical configuration at any stereocenter of a complex represented by A is R, S, or a mixture of these configurations.
  • the present invention relates to a complex represented by A and the attendant definitions, wherein X represents O.
  • the present invention relates to a complex represented by A and the attendant definitions, wherein Z represents a chelator selected from the group consisting of bis(2-pyridylmethyl)ammes, N-pyridyl-2-amino-carboxylic acids, N x S y chelators, wherein x is 1, 2, 3, or 4, and y is 0, 1, 2, 3, or 4, DTPA, EDTA, SHNH, and mercaptoacetyltripeptides.
  • the present mvention relates to a complex represented by A and the attendant definitions, wherem Z represents a chelator selected ftom the group consisting of bis(2-pyridylmethyl)amines, N-(2-pyridylmethyl)-2-amino-carboxylic acids, and N x S y chelators, wherem x is 1, 2, 3, or 4, y is 0, 1, 2, or 3, and the sum of x and y is 4.
  • the present invention relates to a complex represented by A and the attendant definitions, wherein Z represents a chelator selected from the group consisting of bis(2-pyridylmethyl)amine, N-(2-pyridylmethyl)glycine, and N 2 S 2 chelators.
  • the present mvention relates to a complex represented by A and the attendant definitions, wherein said radionuclide is technetium.
  • the present invention relates to a complex represented by A and the attendant definitions, wherein R represents H.
  • the present invention relates to a complex represented by A and the attendant definitions, wherein R' represents methyl.
  • the present invention relates to a complex represented by A and the attendant definitions, wherein R" represents H. In certain embodiments, the present invention relates to a complex represented by A and the attendant definitions, wherein R 3 represents H.
  • the present invention relates to a complex represented by A and the attendant definitions, wherein the stereochemical relationship between the two instances of R 3 is cis.
  • the present invention relates to a complex represented by A and the attendant definitions, wherein R 3 represents H; and the stereochemical relationship between the two instances of R 3 is cis.
  • the present invention relates to a complex represented by A and the attendant definitions, wherein R represents H; R' represents methyl; R" represents H; R 3 represents H; and the stereochemical relationship between the two instances of R 3 is cis.
  • the present invention relates to a complex represented by A and the attendant definitions, wherein X represents O; and Z represents a chelator selected from the group consisting of bis(2-pyridylmethyl)amines, N-pyridyl-2-amino-carboxylic acids, N x S y chelators, wherein x is 1, 2, 3, or 4, and y is 0, 1, 2, 3, or 4, DTPA, EDTA,
  • the present invention relates to a complex represented by A and the attendant definitions, wherein X represents O; and Z represents a chelator selected from the group consisting of bis(2-pyridylmethyl)amines, N-(2-pyridylmethyl)-2-amino- carboxylic acids, and N x S y chelators, wherein x is 1, 2, 3, or 4, y is 0, 1, 2, or 3, and the sum of x and y is 4.
  • the present invention relates to a complex represented by A and the attendant definitions, wherein X represents O; and Z represents a chelator selected from the group consisting of bis(2-pyridylmethyl)amine, N-(2-pyridylmemyl)glycine, and N 2 S 2 chelators.
  • the present invention relates to a complex represented by A and the attendant definitions, wherein X represents O; Z represents a chelator selected from the group consisting of bis(2-pyridylmethyl)amines, N-pyridyl-2-amino-carboxylic acids, N x S y chelators, wherein x is 1, 2, 3, or 4, and y is 0, 1, 2, 3, or 4, DTPA, EDTA, SHNH, and mercaptoacetyltripeptides; and said radionuclide is technetium.
  • the present invention relates to a complex represented by A and the attendant definitions, wherein X represents O; Z represents a chelator selected from the group consisting of bis(2-pyridylmethyl)amines, N-(2-pyridylmethyl)-2-amino- carboxylic acids, and N x S y chelators, wherein x is 1, 2, 3, or 4, y is 0, 1, 2, or 3, and the sum of x and y is 4; and said radionuclide is technetium.
  • the present invention relates to a complex represented by A and the attendant definitions, wherein X represents O; Z represents a chelator selected from the group consisting of bis(2-pyridylmethyl)amine, N-(2-pyridylmethyl)glycine, and N 2 S 2 chelators; and said radionuclide is technetium.
  • the present invention relates to a complex represented by A and the attendant definitions, wherein X represents O; Z represents a chelator selected from the group consisting of bis(2-pyridylmethyl)amines, N-pyridyl-2-amino-carboxylic acids, N x S y chelators, wherein x is 1, 2, 3, or 4, and y is 0, 1, 2, 3, or 4, DTPA, EDTA, SHNH, and mercaptoacetyltripeptides; said radionuclide is technetium; R represents H; R' represents methyl; R" represents H; R 3 represents H; and the stereochemical relationship between the two instances of R 3 is cis.
  • X represents O
  • Z represents a chelator selected from the group consisting of bis(2-pyridylmethyl)amines, N-pyridyl-2-amino-carboxylic acids, N x S y chelators, wherein x is 1, 2, 3, or 4, and y is 0, 1,
  • the present mvention relates to a complex represented by A and the attendant definitions, wherein X represents O; Z represents a chelator selected from the group consisting of bis(2-pyridylmethyl)amines, N-(2-pyridylmethyl)-2-amino- carboxylic acids, and N x S y chelators, wherem x is 1, 2, 3, or 4, y is 0, 1, 2, or 3, and the sum of x and y is 4; said radionuclide is technetium; R represents H; R' represents methyl; R" represents H; R 3 represents H; and the stereochemical relationship between the two instances of R 3 is cis.
  • X represents O
  • Z represents a chelator selected from the group consisting of bis(2-pyridylmethyl)amines, N-(2-pyridylmethyl)-2-amino- carboxylic acids, and N x S y chelators, wherem x is 1, 2, 3, or 4, y is 0, 1, 2, or 3, and the sum
  • the present invention relates to a complex represented by A and the attendant definitions, wherem X represents O; Z represents a chelator selected from the group consisting of bis(2-pyridylmethyl)amine, N-(2-pyridylmethyl)glycine, and N 2 S 2 chelators; said radionuclide is technetium; R represents H; R' represents methyl; R" represents H; R 3 represents H; and the stereochemical relationship between the two instances of R 3 is cis.
  • Another aspect of the present invention relates to a composition, comprising a complex ofthe present invention; and a pharmaceutically acceptable excipient.
  • a third aspect of the present invention relates to a method of imaging a region in a patient, comprising the steps of: administering to a patient a diagnostically effective amount of an complex of the present invention; exposing a region of said patient to radiation; and obtaining an image of said region of said patient.
  • the present invention relates to the aforementioned method, wherein said region of said patient is the thorax.
  • the present invention relates to the aforementioned method, wherein said region of said patient is the heart.
  • a fourth aspect of the present invention relates to a kit, comprising a complex ofthe present invention in a container, and instructions for using said complex to image a region in a patient.
  • the radioisotope technetium-99m with a 6 hour half-life, gamma energy of 140 keV (85% of the gamma photons emit at 140 keV), wide-spread availability, and low cost makes it a preferred choice of radionuclides for nuclear medicine imaging today. Reedijk J. "Medicinal Applications of heavy-metal compounds” Curr. Opin. Chem. Biol. (1999) 3(2): 236-240. These advantages, coupled with the fact that Single Photon Emission Computed Tomography (SPECT) cameras are optimized for the 140 keV energy of Tc-99m, clearly establish the superiority of Tc-99m-labeled imaging agents.
  • SPECT Single Photon Emission Computed Tomography
  • DADT monoamine monoamide
  • PAMA picolinamine mono-acetic acid
  • the DADT, MAMA and PAMA moieties were selected because they can chelate a wide range of technetium complexes, varying in metal oxidation states, size, and lipophilicity.
  • Three series of novel rotenone analogs labeled with Tc-99m at the 7' position were prepared and their chemical structures identified by preparation of the corresponding rhenium analogs.
  • the relationship in the Periodic Table between technetium and rhenium implies that Tc-99m radiopharmaceuticals can be designed by modeling analogous rhenium complexes. Rose, D. J., Maresca, K.
  • This stable core has the additional advantage of being smaller and less polar than the routinely employed Tc(V)-oxo systems. This could offer a big advantage in biologically relevant systems where the addition of the metal center effects the size, shape, and potentially the bioactivity ofthe compounds.
  • the non-polar precursor Tc(CO) 3 + with three tightly bound "innocent” carbonyls, provides three open coordination sites, allowing for a large degree of flexibility in the choice of ligands.
  • the picolinamine mono- acetic acid (PAMA) ligand provides both oxygen and nitrogens as potential donor atoms.
  • PAMA picolinamine mono- acetic acid
  • One aspect of the present mvention relates to the design and synthesis of a series of rotenone derivatives, comprising a metal chelator substituent, which may be varied in terms of size and lipophility.
  • Another aspect of the present invention relates to labeling the aforementioned rotenone derivatives with Tc-99m, using labeling methods based on both the Tc(V)-oxo core and Tc(I)(CO) 3 L 3 core.
  • the labeled rotenone analogues are characterized structurally by comparison to the corresponding Re(IV) and Re(I) derivatives.
  • the present invention relates to assessment of the in vivo pharmacokinetic properties in rats of the Tc-99m labeled novel rotenone derivatives, including a comparison to the conesponding pharmacokinetic properties of Tl-201 chloride. Also, a control experiment compares the biodistribution in rats of Tc99m-sestamibi with that of the Tc-
  • kits formulated 99m Tc-cardiac imaging agents The technetium-99m labeled rotenone derivatives have also been assessed for their suitability for inclusion in a kit, including a determination of their stability as a function of time and concentration.
  • the present invention also relates to an examination ofthe stability of these complexes in buffer at physiological pH, and in human plasma and serum components. Identification of an agent that allows for improved noninvasive delineation of myocardial perfusion and which could be routinely prepared at most clinical institutions or purchased from a centralized nuclear pharmacy would be of considerable benefit in the diagnosis and treatment of heart disease and constitute a significant diagnostic and commercial opportunity.
  • Optimum complexes to image myocardial blood flow are being developed, and mechanistic studies are being performed to characterize flow and biochemically related behaviors, and to develop a convenient kit formulation for the optimum Tc-agent.
  • Chelators which bind to radionuclides are known in the art. See, e.g., M. Nicolini et al., eds., "Technetium and Rhenium in Chemistry and Nuclear Medicine," SGEditoriali, Padova (1995).
  • chelators used in the complexes of the present mvention are capable of binding to radionuclides, such as Tc(CO) 3 + or Tc(O) 3+ .
  • a chelator moiety will be a tetradentate chelator, i.e., will be capable of four- point binding to a radionuclide.
  • Exemplary tetradentate chelators include N 2 S 2 and N 3 S chelators, as described in, e.g., A. R. Fritzberg, et al., J. Nucl. Med. 23:592-598 (1982); S. Liu and D. S. Edwards, in M. Nicolini et al., eds., "Technetium and Rhenium in Chemistry and Nuclear Medicine," op. cit, pp. 383-393; and S. Vallabhajousula et al., J. Nucl. Med. 30:599-604 (1989).
  • N 2 S 2 chelator can chelate a radionuclide through two nitrogen atoms (e.g., amido nitrogens of a peptide backbone) and two sulfur atoms (e.g., of a mercaptoacetyl moiety), while N 3 S chelators can chelate to a radionuclide through three nitrogen atoms and one sulfur atom.
  • nitrogen atoms e.g., amido nitrogens of a peptide backbone
  • sulfur atoms e.g., of a mercaptoacetyl moiety
  • N x S y chelating compound includes bifunctional chelators that are capable of: (i) coordmately binding a radionuclide; and (ii) covalently attaching to an rotenone analog.
  • Preferred N x S y chelating compounds have the N 2 S 2 (generally described in U.S. Pat. Nos. 4,897,225 or 5,164,176 or 5,120,526), N 3 S (generally described in U.S. Pat. No. 4,965,392), N 2 S 3 (generally described in U.S. Pat. No. 4,988,496), N 2 S 4 (generally described in U.S. Pat. No.
  • N x S y chelating compounds have N 2 S 2 and N 3 S cores.
  • Exemplary N x S y chelating compounds are described in Fritzberg et al., Proc. Natl. Acad. Sci. USA 85:4024-29, 1988; in Weber et al., Bioconj. Chem. 1:431-37, 1990; and in the references cited therein.
  • the N 2 S 2 chelating compounds include diamide, dimercaptide bifunctional chelators of the N x S y family capable of stably complexing a radionuclide through two nitrogen atoms and two sulfur atoms that are appropriately positioned.
  • N 2 S 2 chelating compounds are generally described in U.S. Pat. No. 4,897,225.
  • the N 3 S chelatmg compounds include triamide, mercaptide bifunctional chelators of the N x S y family capable of stably complexing a radionuclide through three nitrogen atoms and one sulfur atom that are appropriately positioned.
  • Preferred N 3 S chelating compounds are described in U.S. Pat. Nos. 4,965,392 and 5,091 ,514.
  • chelating agents include, for example, diethylene triamine pentaacetic acid (DTPA) and ethylene diamine tetracetic acid (EDTA).
  • DTPA diethylene triamine pentaacetic acid
  • EDTA ethylene diamine tetracetic acid
  • Other chelators appropriate to link a radionuclide to a compound in accordance with the present invention are described in standard texts such as Advanced Inorganic Chemistry, 4th edition, 1980, F. A. Cotton and G. Wilkinson, John Wiley & Sons.
  • the most suitable metal chelating agent will vary with the metal to be chelated, e.g. depending on its particular coordination geometry.
  • Chelators suitable specifically for linking 99m Tc to rotenone compounds in accordance with the present invention preferably present, as a metal coordinating configuration, a combination of four nitrogen and sulfur metal-coordinating atoms.
  • proteins have been labeled with technetium-99m ( 99m Tc) using the hydrazino nicotinamide (SHNH) chelator (Abrams M. J., Juweid M., tenKate C. I., Schwartz D. A., Hauser M. M., Gaul F. E., Fuccello A. J., Rubin R. H., Strauss H. W., Fischman A. J, J. Nucl. Med., 31 : 2022-2028, 1990), and the label was found to be stable both in vitro and in vivo (Hnatowich D.
  • SHNH hydrazino nicotinamide
  • the SHNH chelator was initially used for oligonucleotides, however, transfer of label nonspecifically to proteins from oligonucleotides labeled in this manner was observed.
  • the identical oligonucleotide; radiolabeled with In-I l l using the chelator diethylenetriamine-pentaacetic acid (DTPA) showed no tendency to bind to serum proteins under circumstances in which the 99m Tc- SHNH-labeled oligonucleotide was largely protein bound (Hnatowich D.
  • 99m Tc was originally developed as an alternative to radiolabeled hippuran for renal function studies (Fritzberg A R., Kasina S., Eshima D., Johnson D. L., J. Nucl. Med., 27: 111-116; 1986).
  • This succinimide ester mercapto-acetyl tripeptide is protected against disulfide-bond formation by a benzoyl group, which must be heated to 100 C for 10 min during labeling to remove the protecting group.
  • This benzoyl-protected chelator has also been used to radiolabel antibodies with 99m Tc (Fritzberg A. R., Berninger R. W., Hadley S. W.
  • prefened chelator moieties include amidothiols, including, e.g., mercaptoacetyltripeptides, such as, e.g., mercaptoacetyltriglycine (MAG 3 ), mercaptoacetyltriserine.
  • mercaptoacetyl-tripeptides can chelate radionuclides, such as Tc(CO) 3 + or Tc(O) 3+ , by coordination through the three amide nitrogens of the peptide backbone, and the terminal mercapto group.
  • Other chelator moieties which may find use in the present invention include cyclams, porphyrins, crown ethers, azacrown ethers, and the like. Linking Moiety
  • the chelator is bonded directly to the methylene carbon of the isopropylidene group at the 2-position of the dihydrofuran nucleus of rotenone.
  • the chelator and the rotenone core of the complexes of the present invention are connected by a linker moiety, i.e., a covalent tether.
  • Linking moieties provided by the invention are comprised of at least two linker functional groups each capable of covalently bonding to the rotenone core and the chelator.
  • Such functional groups include but are not limited to primary and secondary amines, hydroxyl groups, carboxylic acid groups and thiol groups.
  • a rotenone analog may comprise a linker that serves to create a physical separation between the chelator and the rotenone.
  • a linker may be an alkyl chain that is derivatized for coupling to the chelator.
  • the spacer may comprise one or more amino acid residues.
  • the chelator may be linked to the rotenone core in a variety of ways.
  • the carboxyl group may be activated with carbodiimide and an alcohol to form an active ester or may already contain an active ester group that is reacted with an available amino group of a polypeptide or an amino sugar to form an amide bond.
  • the amino group may be used to react with an aldehyde group, which may be derived by glycol cleavage of a sugar with periodate, to form a Schiff s base or cyclic imine or under conditions favoring reductive amination to form a secondary or tertiary amine or cyclic imine linkage.
  • an aldehyde group which may be derived by glycol cleavage of a sugar with periodate, to form a Schiff s base or cyclic imine or under conditions favoring reductive amination to form a secondary or tertiary amine or cyclic imine linkage.
  • the chelated radionuclide is bound to the bio-molecule via a pendant chain distant to the receptor-binding site.
  • chelators are currently employed in the binding of tectnetium, all of these tracers suffer from one or more disadvantages which render them less than ideal: HYNIC requires coligands; MAG3 may be only used with the Tc(V)-oxo species; EDTA/DTPA is used primarily primarily with Tc(V)-oxo and its ability to retain label is poor. Hence, additional Technetium-99m chelators are needed. Novel radiolabeled chelators that display rapid, efficient labeling and demonstrate superior labeling retention for both Tc(V)-oxo and Tc(I)-tricarbonyl cores without the use of coligands are attractive candidates for clinical evaluation as potential chelators for biologically relevant molecules.
  • Using the neutral metal chelate analogs has a number of advantages: a) there is no charge change and the labeled molecule is expected to retain the membrane diffusional properties of rotenone, b) these chelators have proven to be good ligands for binding Tc- 99m at room temperature in high radiochemical yields and purity, c) the ligands keep the metal in a thermodynamically stable +5 or +1 oxidation state, and d) the size of the Tc- 99m-chelate is similar to that of the phenyl group, which should not perturb the mitochondria binding system. Warren GL, Caldwell JH, Kremer PA, et al. "New iodinated phenyl fatty acids for imaging myocardial metabolism" J. Nucl. Med. (1986) 939-940. Two preferred derivatives include conjugation through one of the amines of the diaminodithiol (DADT) chelate (1) or the amine of the monoamine monoamide (MAMA)
  • a second embodiment exemplified by analog 3, incorporates the technetium (I) tricarbonyl center capped with a coordinating picolinamine mono-acetic acid moiety (PAMA).
  • PAMA picolinamine mono-acetic acid moiety
  • All compounds of the present invention are also synthesized with macroscopic quantities of rhenium for characterization by conventional spectral and elemental analyses, fast atom bombardment mass spectrometry, l H and 13 C NMR spectrometry, and infrared analysis. Following purification, all non-radioactive compounds are analyzed for chemical purity by elemental analysis, thin-layer and high-pressure liquid chromatography. X-ray crystallography may also be performed on rhenium analogs. All radiolabeled agents are analyzed for radio-homogeneity by thin-layer and high-pressure liquid chromatography.
  • the rotenone-MAMA chelator 2 has been prepared as shown below.
  • rotenone-picolinamine mono-acetic acid (PAMA) chelator 3 has been prepared as shown below.
  • Tc-99m-labeled rotenone derivatives is achieved by adding 10 mCi of TcO 4 " to a 0.9% saline solution of sodium gluceptate (200 mg/3 mL). After 20 minute incubation, 400 ul is added to a solution of 400 ul of sodium acetate (50 mM, pH 5.2) and the appropriate rotenone N 2 S 2 derivative (50 ug). The mixture is heated at 80 °C for 30 min. The mixture is then extracted with ethyl acetate (3 x 1 mL), dried over sodium sulfate, and dried under N 2 . ⁇ ie residue is then re-dissolved in ethanol (400 ul) and purity checked via HPLC by a Vydac C18 (5 mm, 25 cm) column using methanol to elute the reaction products.
  • the Tc(I) carbonyl chemistry allows for the possibility of an alternative route to form stable 99m Tc-rotenone complexes.
  • Na 2 CO 3 0.004 g, 0.038 mmol
  • NaBH 4 0.005 g, 0.13 mmol
  • the vial is sealed and flushed with CO for 10 min.
  • To the vial is added 1 mL of Na 99m Tc0 4 " in saline.
  • the solution is heated to 75° C for 30 minutes.
  • 0.3ml of 1M PBS solution is added (pH 7.4), resulting in the stable formation of [ 99m Tc(OH 2 ) 3 (CO) 3 ] + .
  • This Tc(I) tricarbonyl species is then heated at 75° C for 30 minutes with the PAMA derivatized rotenone to form the 99m Tc-rotenone complex.
  • a 'one pot' synthesis has also been performed where the PAMA derivatized rotenone is added to the vial with the Na 2 CO 3 , NaBH 4 , and flushed with CO for 10 min. After the flush, 1 mL of Na 99m TcO 4 " in saline is added. Finally the solution is heated to 75° C for 30 minutes. The reaction is then checked for purity via HPLC by a Vydac C18 (5 mm, 25 cm) column using methanol to elute the reaction products. Synthesis of rhenium analogs for structural characterization
  • the properties ofthe Group VII metals technetium and rhenium are very similar due to their periodic relationship. Therefore, the metals typically demonstrate similar reaction chemistry, for example, the thiol, nitrogen, phosphine and oxo-chemistry of these two metals. Likewise, penhenate and pertechnetate have very similar chemistries. Rose, D. J., Maresca, K. P., Nicholson, T., Davison, A., Jones, A. G., Babich, J. ; Fischman, A., Graham, W., DeBord, J. R. D., Zubieta, J.
  • Synthesizing the rhenium-rotenone complexes provides a facile route to characterize structurally the products.
  • the characterized products lead to the development of new Tc-rotenone derivatives based on the presence or absence of a structural feature found in the rhenium data obtained.
  • the periodic relationship between Tc and Re indicates that Tc-99m radiopharmaceuticals can be designed by modeling analogous rhenium complexes. Nicholson, T., Cook, J., Davison, A., Rose, D. J., Maresca K. P., Zubieta, J. A., Jones, A. G.
  • Re(V) complexes The synthesis of the Re(V) complexes is accomplished by reacting [TBA][ReOBr 4 (OPPh 3 )j with the appropriate rotenone ligand in the ratio of 1: 1.2 in 10 mL of methanol and three equivalents of NEt 3 as base. The reaction is then allowed to reflux for l A hour. After cooling the reaction products are purified using a small column using the method established by Spies and co-workers. Spies, H., Fietz, T., Glaser, M., Pietzsch, H.-J., Johannsen, B.
  • the rhenium (V) starting material [ReOCl 3 (PPh 3 ) 2 ] may be employed as the potential rhenium starting material.
  • This versatile material has proven successful in the past in dealing with nitrogen and sulfur donor atoms. Maresca, K. P., Femia, F. J., Bonavia, G. H., Babich, J. W., Zubieta, J.
  • the Re(I)(CO) 3 + system follows similar reaction chemistry to that of the Tc-99m tricarbonyl core.
  • the use of [NEt 4 ] 2 [ReBr 3 (CO) 3 ], as the starting material leads to easy formation of the_ ⁇ c-Re(CO) 3 (L) 3 core.
  • the [NEt 4 ] 2 [ReBr 3 (CO) 3 ] is readily derived from the
  • Vertebrate animals in this research project are used to investigate the biodistribution and pharmacokinetics of new technetium-rotenone complexes and determine their uptake in the heart.
  • Rats (Sprague Dawley, male, at 80-100 grams each) are used for the whole body biodistribution studies. Three compounds are evaluated in this series with five time points 5, 15, 30, 60, and 120 minutes with five animals per time point. This many animals are needed to provide accurate statistics in the clearance rate measurements and to account for intraspecies variation.
  • the stability of the radiolabeled compounds in solution and in plasma is determined as a function of time and solution conditions such as pH and solvents. Specifically, after radiolabeling and isolation, the product is allowed to sit at room temperature for 48 hours, after which HPLC analysis is performed to check for degree of label retention, as well as potential product degradation. We then analyze for the reformation of TcO 4 " and the presence of the reduced species Tc0 2 . To assist in predicting the in vivo label stability, we perform ligand challenges. Specifically, we incubate the product with a competing biological ligand, such as cysteine, albumin, and transferrin, testing the stability of the radiolabel via HPLC analysis. Finally we test the product in plasma as a function of time and pH. Development of a convenient kit formulation for the agent with the highest heart specificity and selectivity
  • Tc-99m The widespread availability of Tc-99m from a generator has made it the most frequently used nuclide in nuclear medicine.
  • the advantage of a Tc-99m labeled rotenone complex is that it allows for the possible development of a convenient "kit” formulation, thereby permitting more accessibility of the diagnostic agent.
  • 99m Tc-rotenone complexes can be synthesized in a "one-step” procedure.
  • the appropriate coligand, as well as the conect stoichiometric amounts of reducing agents and coligands are being developed. Both the Tc(V)-oxo system and the Tc(I)-carbonyl system possess the potential to be routes to 99m Tc-rotenone kits.
  • N 2 S 2 and PAMA as chelates for the 99m Tc we have chosen stable cores with saturated metal binding sites, limiting the number of possible side-products.
  • the potential to prepare the compound into a kit lies in the chelates' ability to bind rapidly and securely to the metal.
  • kits containing the labeled compounds ofthe present invention.
  • kits may contain the labeled compounds in sterile lyophilized form, and may include a sterile container of a radiopharmaceutically acceptable reconstitution liquid. Suitable reconstitution liquids are disclosed in Remington's Pharmaceutical Sciences and The United States Pharmacopia ⁇ The National Formulary, cited above.
  • kits may alternatively contain a sterile container of a composition of the radiolabeled compounds of the invention.
  • kits may also include, if desired, other conventional kit components, such as, for example, one or more carriers, one or more additional vials for mixing.
  • kits for the preparation of radiopharmaceuticals.
  • Diagnostic kits of the present invention comprise one or more vials containing the sterile, non-pyrogenic, formulation comprised of a predetermined amount of a rotenone analog, a reducing agent, and optionally a solubilization aid or other components such as transfer ligands, buffers, lyophilization aids, stabilization aids, and bacteriostats.
  • a solubilization aid or other components such as transfer ligands, buffers, lyophilization aids, stabilization aids, and bacteriostats.
  • the inclusion of one or more optional components in the formulation will frequently improve the ease of synthesis ofthe radiopharmaceutical by the practising end user, the ease of manufacturing the kit, the shelf-life of the kit, or the stability and shelf-life of the radiopharmaceutical.
  • the improvement achieved by the inclusion of an optional component in the formulation must be weighed against the added complexity of the formulation and added cost to manufacture the kit.
  • the one or more vials that contain all or part of the formulation can independently be in the form of a sterile solution or a lyophilized solid.
  • Solubilization aids useful in the preparation of radiopharmaceuticals and in diagnostic kits useful for the preparation of said radiopharmaceuticals include but are not limited to ethanol, glycerin, polyethylene glycol, propylene glycol, polyoxyethylene sorbitan monooleate, sorbitan monoloeate, polysorbates, poly(oxyethylene)poly(oxypropylene)poly(oxyethylene) block copolymers (Pluronics) and lecithin.
  • Prefened solubilizing aids are polyethylene glycol, and Pluronics.
  • Buffers useful in the preparation of radiopharmaceuticals and in diagnostic kits useful for the preparation of said radiopharmaceuticals include but are not limited to phosphate, citrate, sulfosalicylate, and acetate. A more complete list can be found in the United States Pharmacopeia. Lyophilization aids useful in the preparation diagnostic kits useful for the preparation of radiopharmaceuticals include but are not limited to mannitol, lactose, sorbitol, dextran, Ficoll, and polyvinylpyrrolidine(PVP).
  • Stabilization aids useful in the preparation of radiopharmaceuticals and in diagnostic kits useful for the preparation of said radiopharmaceuticals include but are not limited to ascorbic acid, cysteine, monothioglycerol, sodium bisulfite, sodium metabisulfite, gentisic acid, and inositol.
  • Bacteriostats useful in the preparation of radiopharmaceuticals and in diagnostic kits useful for the preparation of said radiopharmaceuticals include but are not limited to benzyl alcohol, benzalkonium chloride, chlorobutanol, and methyl, propyl or butyl paraben.
  • a component in a diagnostic kit can also serve more than one function.
  • a reducing agent can also serve as a stabilization aid, a buffer can also serve as a transfer ligand, a lyophilization aid can also serve as a transfer, ancillary or co-ligand and so forth.
  • the predetermined amounts of each component in the formulation are determined by a variety of considerations that are in some cases specific for that component and in other cases dependent on the amount of another component or the presence and amount of an optional component.
  • the minimal amount of each component is used that will give the desired effect of the formulation.
  • the desired effect of the formulation is that the practicing end user can synthesize the radiopharmaceutical and have a high degree of certainty that the radiopharmaceutical can be safely injected into a patient and will provide diagnostic information about the disease state of that patient.
  • the diagnostic kits of the present invention also contain written instructions for the practicing end user to follow to synthesize the radiopharmaceuticals. These instructions may be affixed to one or more of the vials or to the container in which the vial or vials are packaged for shipping or may be a separate insert, termed the package insert.
  • chelator refers to a moiety that is capable of binding a radionuclide, preferably through non-covalent interactions, e.g., through ionic interactions.
  • Chelator moieties suitable for use in the compositions and methods of the invention are preferably capable of binding to a radionuclide with a high affinity, e.g., a binding affinity sufficiently high to permit binding of a radionuclide, preferably under physiological conditions, e.g., in vivo.
  • linking group refers to a chemical group that serves to couple the rotenone analog to the chelator while not adversely affecting either the targeting function of the rotenone analog or the metal binding function of the chelator.
  • Suitable linking groups include alkyl chains; alkyl chains optionally substituted with one or more substituents and in which one or more carbon atoms are optionally replaced with nitrogen, oxygen or sulfur atoms.
  • linking groups include those having the formula A 1 - A 2 -A 3 , wherein A 1 and A 3 are independently selected from N, O and S; and A 2 includes alkyl optionally substituted with one or more substituents, and in which one or more carbon atoms are optionally replaced with nitrogen, oxygen or sulfur atoms; aryl optionally substituted with one or more substituents; and heteroaryl optionally substituted with one or more substituents.
  • Still other suitable linking groups include amino acids and amino acid chains functionalized with one or more reactive groups for coupling to the rotenone analog and/or chelator.
  • the term "diagnostic kit,” as used herein, comprises a collection of components, termed the fonnulation, in one or more vials which are used by the practising end user in a clinical or pharmacy setting to synthesize the radiopharmaceutical.
  • the kit provides all the requisite components to synthesize and use the radiopharmaceutical except those that are commonly available to the practising end user, such as water or saline for injection, a solution of the radionuclide, equipment for heating the kit during the synthesis of the radiopharmaceutical if required, equipment necessary for administering the radiopharmaceutical to the patient such as syringes and shielding, and imaging equipment.
  • heteroatom as used herein means an atom of any element other than carbon or hydrogen. Preferred heteroatoms are boron, nitrogen, oxygen, phosphorus, sulfur and selenium.
  • electron- withdrawing group is recognized in the art, and denotes the tendency of a substitaent to attract valence electrons from neighboring atoms, i.e., the substitaent is electronegative with respect to neighboring atoms.
  • a quantification of the level of electron- withdrawing capability is given by the Hammett sigma ( ⁇ ) constant. This well known constant is described in many references, for instance, J. March, Advanced Organic Chemistry, McGraw Hill Book Company, New York, (1977 edition) pp. 251-259.
  • Exemplary electron-withdrawing groups include nitro, acyl, formyl, sulfonyl, trifluoromethyl, cyano, chloride, and the like.
  • Exemplary electron- donating groups include amino, methoxy, and the like.
  • alkyl refers to the radical of saturated aliphatic groups, including straight-chain alkyl groups, branched-chain alkyl groups, cycloalkyl (alicyclic) groups, alkyl substituted cycloalkyl groups, and cycloalkyl substituted alkyl groups.
  • a straight chain or branched chain alkyl has 30 or fewer carbon atoms in its backbone (e.g., C1-C30 for straight chain, C3-C30 for branched chain), and more preferably 20 or fewer.
  • prefened cycloalkyls have from 3-10 carbon atoms in their ring structure, and more preferably have 5, 6 or 7 carbons in the ring structure.
  • lower alkyl as used herein means an alkyl group, as defined above, but having from one to ten carbons, more preferably from one to six carbon atoms in its backbone structure. Likewise, “lower alkenyl” and “lower alkynyl” have similar chain lengths. Prefened alkyl groups are lower alkyls. In prefened embodiments, a substitaent designated herein as alkyl is a lower alkyl.
  • aralkyl refers to an alkyl group substituted with an aryl group (e.g., an aromatic or heteroaromatic group).
  • alkenyl and alkynyl refer to unsaturated aliphatic groups analogous in length and possible substitution to the alkyls described above, but that contain at least one double or triple bond respectively.
  • aryl as used herein includes 5-, 6- and 7-membered single-ring aromatic groups that may include from zero to four heteroatoms, for example, benzene, pynole, furan, thiophene, imidazole, oxazole, thiazole, triazole, pyrazole, pyridine, pyrazine, pyridazine and pyrimidine, and the like.
  • aryl groups having heteroatoms in the ring structare may also be refened to as "aryl heterocycles" or "heteroaromatics.”
  • the aromatic ring can be substituted at one or more ring positions with such substituents as described above, for example, halogen, azide, alkyl, aralkyl, alkenyl, alkynyl, cycloalkyl, hydroxyl, alkoxyl, amino, nitro, sulfhydryl, imino, amido, phosphonate, phosphinate, carbonyl, carboxyl, silyl, ether, alkylthio, sulfonyl, sulfonamido, ketone, aldehyde, ester, heterocyclyl, aromatic or heteroaromatic moieties, -CF3, -CN, or the like.
  • aryl also includes polycyclic ring systems having two or more cyclic rings in which two or more carbons are common to two adjoining rings (the rings are "fused rings") wherein at least one ofthe rings is aromatic, e.g., the other cyclic rings can be cycloalkyls, cycloalkenyls, cycloalkynyls, aryls and/or heterocyclyls.
  • ortho, meta and para apply to 1,2-, 1,3- and 1 ,4-disubstituted benzenes, respectively.
  • the names 1,2-dimethylbenzene and ort/.o-dimethylbenzene are synonymous.
  • heterocyclyl or “heterocyclic group” refer to 3- to 10-membered ring structures, more preferably 3- to 7-membered rings, whose ring structures include one to four heteroatoms. Heterocycles can also be polycycles.
  • Heterocyclyl groups include, for example, azetidine, azepine, thiophene, thianthrene, furan, pyran, isobenzofuran, chromene, xanthene, phenoxathiin, pynole, imidazole, pyrazole, isothiazole, isoxazole, pyridine, pyrazine, pyrimidine, pyridazine, indolizine, isoindole, indole, indazole, purine, quinolizine, isoquinoline, quinoline, phthalazine, naphthyridine, quinoxaline, quinazoline, cinnoline, pteridine, carbazole, carboline, phenanthridine, acridine, pyrimidine, phenanthroline, phenazine, phenarsazine, phenothiazine, furazan,
  • the heterocyclic ring can be substituted at one or more positions with such substituents as described above, as for example, halogen, alkyl, aralkyl, alkenyl, alkynyl, cycloalkyl, hydroxyl, amino, nitro, sulfhydryl, imino, amido, phosphonate, phosphinate, carbonyl, carboxyl, silyl, ether, alkylthio, sulfonyl, ketone, aldehyde, ester, a heterocyclyl, an aromatic or heteroaromatic moiety, -CF3, -CN, or the like.
  • substituents as described above, as for example, halogen, alkyl, aralkyl, alkenyl, alkynyl, cycloalkyl, hydroxyl, amino, nitro, sulfhydryl, imino, amido, phosphonate, phosphinate, carbonyl, carboxyl,
  • polycyclyl or “polycyclic group” refer to two or more rings (e.g., cycloalkyls, cycloalkenyls, cycloalkynyls, aryls and/or heterocyclyls) in which two or more carbons are common to two adjoining rings, e.g., the rings are "fused rings". Rings that are joined through non-adjacent atoms are termed "bridged" rings.
  • Each of the rings of the polycycle can be substituted with such substituents as described above, as for example, halogen, alkyl, aralkyl, alkenyl, alkynyl, cycloalkyl, hydroxyl, amino, nitro, sulfhydryl, imino, amido, phosphonate, phosphinate, carbonyl, carboxyl, silyl, ether, alkylthio, sulfonyl, ketone, aldehyde, ester, a heterocyclyl, an aromatic or heteroaromatic moiety, - CF3, -CN, or the like.
  • substituents as described above, as for example, halogen, alkyl, aralkyl, alkenyl, alkynyl, cycloalkyl, hydroxyl, amino, nitro, sulfhydryl, imino, amido, phosphonate, phosphinate, carbonyl, carboxyl, sily
  • carrier refers to an aromatic or non-aromatic ring in which each atom ofthe ring is carbon.
  • nitro means -NO2; the term “halogen” designates -F, -CI, -Br or -I; the term “sulfhydryl” means -SH; the term “hydroxyl” means -OH; and the term “sulfonyl” means -SO2-.
  • amine and “amino” are art-recognized and refer to both unsubstituted and substituted amines, e.g., a moiety that can be represented by the general formula:
  • R9, Ri g and R'10 each independently represent a group permitted by the rules of valence.
  • acylamino is art-recognized and refers to a moiety that can be represented by the general formula:
  • Rg is as defined above, and R'i ]_ represents a hydrogen, an alkyl, an alkenyl or -(CH2) m -R8, where m and Rg are as defined above.
  • amino is art recognized as an amino-substitated carbonyl and includes a moiety that can be represented by the general formula:
  • Prefened embodiments of the amide will not include imides which may be unstable.
  • alkylthio refers to an alkyl group, as defined above, having a sulfur radical attached thereto.
  • the "alkylthio" moiety is represented by one of -S-alkyl, -S-alkenyl, -S-alkynyl, and -S-(CH2) m -R8, wherein m and Rg are defined above.
  • Representative alkylthio groups include methylthio, ethyl thio, and the like.
  • carbonyl is art recognized and includes such moieties as can be represented by the general formula:
  • X is a bond or represents an oxygen or a sulfur
  • Rj j represents a hydrogen, an alkyl, an alkenyl, -(CH2) - or a pharmaceutically acceptable salt
  • R'n represents a hydrogen, an alkyl, an alkenyl or -(CH2) m -Rg, where m and Rg are as defined above.
  • X is an oxygen and R ⁇ or R'n is not hydrogen
  • the formula represents an "ester”.
  • X is an oxygen, and R ⁇ ⁇ is as defined above, the moiety is referred to herein as a carboxyl group, and particularly when RJ is a hydrogen, the formula represents a "carboxylic acid".
  • alkoxyl or "alkoxy” as used herein refers to an alkyl group, as defined above, having an oxygen radical attached thereto.
  • Representative alkoxyl groups include methoxy, ethoxy, propyloxy, tert-butoxy and the like.
  • An "ether” is two hydrocarbons covalently linked by an oxygen. Accordingly, the substitaent of an alkyl that renders that alkyl an ether is or resembles an alkoxyl, such as can be represented by one of -O-alkyl, -O- alkenyl, -O-alkynyl, -0-(CH2)m"R8» where m and Rg are described above.
  • R41 is an electron pair, hydrogen, alkyl, cycloalkyl, or aryl.
  • triflyl, tosyl, mesyl, and nonaflyl are art-recognized and refer to trifluoromethanesulfonyl, /j-toluenesulfonyl, methanesulfonyl, and nonafluorobutanesulfonyl groups, respectively.
  • triflate, tosylate, mesylate, and nonaflate are art-recognized and refer to trifluoromethanesulfonate ester, /?-toluenesulfonate ester, methanesulfonate ester, and nonafluorobutanesulfonate ester functional groups and molecules that contain said groups, respectively.
  • Me, Et, Ph, Tf, Nf, Ts, Ms represent methyl, ethyl, phenyl, trifluoromethanesulfonyl, nonafluorobutanesulfonyl, j->-toluenesulfonyl and methanesulfonyl, respectively.
  • a more comprehensive list of the abbreviations utilized by organic chemists of ordinary skill in the art appears in the first issue of each volume of the Journal of Organic Chemistry; this list is typically presented in a table entitled Standard List of Abbreviations. The abbreviations contained in said list, and all abbreviations utilized by organic chemists of ordinary skill in the art are hereby incorporated by reference.
  • sulfonyl refers to a moiety that can be represented by the general formula:
  • R44 is selected from the group consisting of hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, or heteroaryl.
  • sulfoxido refers to a moiety that can be represented by the general formula: 0
  • R44 is selected from the group consisting of hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aralkyl, or aryl.
  • a “selenoalkyl” refers to an alkyl group having a substitated seleno group attached thereto.
  • Exemplary “selenoethers” which may be substituted on the alkyl are selected from one of -Se-alkyl, -Se-alkenyl, -Se-alkynyl, and -Se-(CH2) m -R7, m and R7 being defined above.
  • Analogous substitutions can be made to alkenyl and alkynyl groups to produce, for example, aminoalkenyls, aminoalkynyls, amidoalkenyls, amidoalkynyls, iminoalkenyls, iminoalkynyls, thioalkenyls, thioalkynyls, carbonyl-substituted alkenyls or alkynyls.
  • each expression e.g. alkyl, m, n, etc., when it occurs more than once in any structare, is intended to be independent of its definition elsewhere in the same structare.
  • substitution or “substituted with” includes the implicit proviso that such substitution is in accordance with permitted valence of the substituted atom and the substitaent, and that the substitution results in a stable compound, e.g., which does not spontaneously undergo transformation such as by rearrangement, cyclization, elimination, etc.
  • the term "substitated" is contemplated to include all permissible substituents of organic compounds.
  • the permissible substitaents include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and nonaromatic substitaents of organic compounds.
  • Illustrative substitaents include, for example, those described herein above.
  • the permissible substitaents can be one or more and the same or different for appropriate organic compounds.
  • the heteroatoms such as nitrogen may have hydrogen substituents and/or any permissible substituents of organic compounds described herein which satisfy the valences of the heteroatoms. This invention is not intended to be limited in any manner by the permissible substitaents of organic compounds.
  • protecting group means temporary substituents which protect a potentially reactive functional group from undesired chemical transformations.
  • protecting groups include esters of carboxylic acids, silyl ethers of alcohols, and acetals and ketals of aldehydes and ketones, respectively.
  • the field of protecting group chemistry has been reviewed (Greene, T.W.; Wuts, P.G.M. Protective Groups in Organic Synthesis, 2 nd ed.; Wiley: New York, 1991).
  • Certain compounds of the present invention may exist in particular geometric or stereoisomeric forms.
  • the present invention contemplates all such compounds, including cis- and tr-w ⁇ -isomers, R- and S-enantiomers, diastereomers, (D)-isomers, (L)-isomers, the racemic mixtures thereof, and other mixtures thereof, as falling within the scope of the invention. Additional asymmetric carbon atoms may be present in a substitaent such as an alkyl group. All such isomers, as well as mixtures thereof, are intended to be included in this invention.
  • a particular enantiomer of a compound of the present invention may be prepared by asymmetric synthesis, it may be isolated using chiral chromatography methods, or by derivation with a chiral auxiliary, where the resulting diastereomeric mixture is separated and the auxiliary group cleaved to provide the pure desired enantiomers.
  • the molecule contains a basic functional group, such as amino, or an acidic functional group, such as carboxyl
  • diastereomeric salts are formed with an appropriate optically-active acid or base, followed by resolution of the diastereomers thus formed by fractional crystallization or chromatographic means well known in the art, and subsequent recovery ofthe pure enantiomers.
  • Contemplated equivalents of the compounds described above include compounds which otherwise correspond thereto, and which have the same general properties thereof (e.g., functioning as analgesics), wherein one or more simple variations of substituents are made which do not adversely affect the efficacy of the compound in binding to opioid receptors.
  • the compounds of the present invention may be prepared by the methods illustrated in the general reaction schemes as, for example, described below, or by modifications thereof, using readily available starting materials, reagents and conventional synthesis procedures. In these reactions, it is also possible to make use of variants which are in themselves known, but are not mentioned here.
  • the chemical elements are identified in accordance with the Periodic Table of the Elements, CAS version, Handbook of Chemistry and Physics, 67th Ed., 1986-87, inside cover.
  • a complex ofthe present invention is represented by A:
  • X represents independently for each occurrence O or S;
  • Z represents a chelator comprising a radionuclide
  • R represents independently for each occurrence H, lower alkyl, or halogen
  • R' represents independently for each occurrence lower alkyl
  • R" represents independently for each occurrence H, or lower alkyl
  • R 3 represents independently for each occurrence H, or lower alkyl
  • the present invention relates to a complex represented by A and the attendant definitions, wherein X represents O.
  • the present mvention relates to a complex represented by A and the attendant definitions, wherein Z represents a chelator selected from the group consisting of bis(2-pyridylmethyl)amines, N-pyridyl-2-amino-carboxylic acids, N x S y chelators, wherein x is 1, 2, 3, or 4, and y is 0, 1, 2, 3, or 4, DTPA, EDTA, SHNH, and mercaptoacetyltripeptides.
  • Z represents a chelator selected from the group consisting of bis(2-pyridylmethyl)amines, N-pyridyl-2-amino-carboxylic acids, N x S y chelators, wherein x is 1, 2, 3, or 4, and y is 0, 1, 2, 3, or 4, DTPA, EDTA, SHNH, and mercaptoacetyltripeptides.
  • the present invention relates to a complex represented by A and the attendant definitions, wherein Z represents a chelator selected from the group consisting of bis(2-pyridylmethyl)amines, N-(2-pyridylmethyl)-2-amino-carboxylic acids, and N x S y chelators, wherein x is 1, 2, 3, or 4, y is 0, 1, 2, or 3, and the sum of x and y is 4.
  • Z represents a chelator selected from the group consisting of bis(2-pyridylmethyl)amines, N-(2-pyridylmethyl)-2-amino-carboxylic acids, and N x S y chelators, wherein x is 1, 2, 3, or 4, y is 0, 1, 2, or 3, and the sum of x and y is 4.
  • the present invention relates to a complex represented by A and the attendant definitions, wherein Z represents a chelator selected from the group consisting ofbis(2-pyridylmethyl)amine, N-(2-pyridylmethyl)glycine, andN 2 S 2 chelators.
  • the present invention relates to a complex represented by A and the attendant definitions, wherein said radionuclide is technetium.
  • the present invention relates to a complex represented by A and the attendant definitions, wherein R represents H.
  • the present invention relates to a complex represented by A and the attendant definitions, wherein R' represents methyl.
  • the present invention relates to a complex represented by A and the attendant definitions, wherein R" represents H. In certain embodiments, the present invention relates to a complex represented by A and the attendant definitions, wherein R 3 represents H.
  • the present invention relates to a complex represented by A and the attendant definitions, wherein the stereochemical relationship between the two instances of R 3 is cis. In certain embodiments, the present invention relates to a complex represented by A and the attendant definitions, wherein R 3 represents H; and the stereochemical relationship between the two instances of R 3 is cis.
  • the present invention relates to a complex represented by A and the attendant definitions, wherein R represents H; R' represents methyl; R" represents H; R 3 represents H; and the stereochemical relationship between the two instances of R 3 is cis.
  • the present invention relates to a complex represented by A and the attendant definitions, wherein X represents O; and Z represents a chelator selected from the group consisting of bis(2-pyridylmethyl)amines, N-pyridyl-2-amino-carboxylic acids, N x S y chelators, wherein x is 1, 2, 3, or 4, and y is 0, 1, 2, 3, or 4, DTPA, EDTA, SHNH, and mercaptoacetyltripeptides.
  • the present invention relates to a complex represented by A and the attendant definitions, wherein X represents O; and Z represents a chelator selected from the group consisting of bis(2-pyridylmethyl)amines, N-(2-pyridylmethyl)-2-amino- carboxylic acids, and N x S y chelators, wherein x is 1, 2, 3, or 4, y is 0, 1, 2, or 3, and the sum of x and y is 4.
  • the present invention relates to a complex represented by A and the attendant definitions, wherein X represents O; and Z represents a chelator selected from the group consisting of bis(2-pyridylmethyl)amine, N-(2-pyridylmethyl)glycine, and N 2 S 2 chelators.
  • the present invention relates to a complex represented by A and the attendant definitions, wherein X represents O; Z represents a chelator selected from the group consisting of bis(2-pyridylmethyl)amines, N-pyridyl-2-amino-carboxylic acids,
  • N x S y chelators wherein x is 1, 2, 3, or 4, and y is 0, 1, 2, 3, or 4, DTPA, EDTA, SHNH, and mercaptoacetyltripeptides; and said radionuclide is technetium.
  • the present invention relates to a complex represented by A and the attendant definitions, wherein X represents O; Z represents a chelator selected from the group consisting of bis(2-pyridylmethyl)amines, N-(2-pyridylmethyl)-2-amino- carboxylic acids, and N x S y chelators, wherein x is 1, 2, 3, or 4, y is 0, 1, 2, or 3, and the sum of x and y is 4; and said radionuclide is technetium.
  • the present invention relates to a complex represented by A and the attendant definitions, wherein X represents O; Z represents a chelator selected from the group consisting of bis(2-pyridylmethyl)amine, N-(2-pyridylmethyl)glycine, and N 2 S 2 chelators; and said radionuclide is technetium.
  • the present invention relates to a complex represented by A and the attendant definitions, wherein X represents O; Z represents a chelator selected from the group consisting of bis(2-pyridylmethyl)amines, N-pyridyl-2-amino-carboxylic acids, N x S y chelators, wherein x is 1, 2, 3, or 4, and y is 0, 1, 2, 3, or 4, DTPA, EDTA, SHNH, and mercaptoacetyltripeptides; said radionuclide is technetium; R represents H; R' represents methyl; R" represents H; R 3 represents H; and the stereochemical relationship between the two instances of R 3 is cis.
  • X represents O
  • Z represents a chelator selected from the group consisting of bis(2-pyridylmethyl)amines, N-pyridyl-2-amino-carboxylic acids, N x S y chelators, wherein x is 1, 2, 3, or 4, and y is 0, 1,
  • the present invention relates to a complex represented by A and the attendant definitions, wherein X represents O; Z represents a chelator selected from the group consisting of bis(2-pyridylmethyl)amines, N-(2-pyridylmethyl)-2-amino- carboxylic acids, and N x S y chelators, wherein x is 1, 2, 3, or 4, y is 0, 1, 2, or 3, and the sum of x and y is 4; said radionuclide is technetium; R represents H; R' represents methyl; R" represents H; R 3 represents H; and the stereochemical relationship between the two instances of R 3 is cis.
  • X represents O
  • Z represents a chelator selected from the group consisting of bis(2-pyridylmethyl)amines, N-(2-pyridylmethyl)-2-amino- carboxylic acids, and N x S y chelators, wherein x is 1, 2, 3, or 4, y is 0, 1, 2, or 3, and the sum of
  • the present invention relates to a complex represented by A and the attendant definitions, wherein X represents O; Z represents a chelator selected from the group consisting of bis(2-pyridylmethyl)amine, N-(2-pyridylmethyl)glycine, and N 2 S 2 chelators; said radionuclide is technetium; R represents H; R' represents methyl; R" represents H; R 3 represents H; and the stereochemical relationship between the two instances of R 3 is cis.
  • the present invention relates to a formulation, comprising a rotenone complex represented by A and the attendant definitions; and a pharmaceutically acceptable excipient.
  • the rotenone complexes of the present invention may be used as radiographic imaging agents.
  • the rotenone complexes of the present invention are prepared by reacting a rotenone-chelator compound with a radionuclide containing solution under radionuclide complex forming reaction conditions.
  • a technetium agent is desired, the reaction is carried out with a pertechnetate solution under technetium 99 m complex forming reaction conditions.
  • the solvent may then be removed by any appropriate means, such as evaporation.
  • the rotenone complexes are then prepared for administration to the patient by dissolution or suspension in a pharmaceutically acceptable vehicle.
  • the present invention also relates to imaging agents containing a radionuclide complex as described above, in an amount sufficient for imaging, together with a pharmaceutically acceptable radiological vehicle.
  • the radiological vehicle should be suitable for injection or aspiration, such as human serum albumin; aqueous buffer solutions, e.g tris(hydromethyl) aminomethane (and its salts), phosphate, citrate, bicarbonate, etc; sterile water; physiological saline; and balanced ionic solutions containing chloride and or dicarbonate salts or normal blood plasma cations such as calcium, potassium, sodium, and magnesium.
  • the concentration of the imaging agent according to the present invention in the radiological vehicle should be sufficient to provide satisfactory imaging, for example, when using an aqueous solution, the dosage is about 1.0 to 50 millicuries.
  • the imaging agent should be administered so as to remain in the patient for about 1 to 3 hours, although both longer and shorter time periods are acceptable. Therefore, convenient ampules containing 1 to 10 mL of aqueous solution may be prepared.
  • Imaging may be carried out in any workable manner, for example by injecting a sufficient amount of the imaging composition to provide adequate imaging and then scanning with a suitable machine, such as a gamma camera.
  • the present invention relates to a method of imaging a region in a patient, comprising the steps of: administering to a patient a diagnostically effective amount of a rotenone complex of the present invention comprising a radionuclide; exposing a region of said patient to radiation; and obtaining an image of said region of said patient.
  • said region of said patient is the thorax.
  • said region of said patient is the heart.
  • the present invention provides pharmaceutically acceptable compositions which comprise a therapeutically-effective amount of one or more of the complexes described above, formulated together with one or more pharmaceutically acceptable carriers (additives) and/or diluents.
  • the pharmaceutical compositions of the present invention may be specially formulated for administration in solid or liquid form, including those adapted for the following: (1) oral administration, for example, drenches (aqueous or non-aqueous solutions or suspensions), tablets, e.g., those targeted for buccal, sublingual, and systemic abso ⁇ tion, boluses, powders, granules, pastes for application to the tongue; (2) parenteral administration, for example, by subcutaneous, intramuscular, intravenous or epidural injection as, for example, a sterile solution or suspension, or sustained-release formulation; (3) topical application, for example, as a cream, ointment, or a controlled-release patch or spray applied to the skin; (4) intravagina
  • terapéuticaally-effective amount means that amount of a compound, material, or composition comprising a compound of the present invention which is effective for producing some desired therapeutic effect in at least a sub-population of cells in an animal at a reasonable benefit/risk ratio applicable to any medical treatment.
  • phrases "pharmaceutically acceptable” is employed herein to refer to those compounds, materials, compositions, and/or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit/risk ratio.
  • pharmaceutically-acceptable carrier means a pharmaceutically-acceptable material, composition or vehicle, such as a liquid or solid filler, diluent, excipient, or solvent encapsulating material, involved in carrying or transporting the subject compound from one organ, or portion ofthe body, to another organ, or portion of the body.
  • a pharmaceutically-acceptable material such as a liquid or solid filler, diluent, excipient, or solvent encapsulating material, involved in carrying or transporting the subject compound from one organ, or portion ofthe body, to another organ, or portion of the body.
  • Each carrier must be “acceptable” in the sense of being compatible with the other ingredients of the formulation and not injurious to the patient.
  • materials which can serve as pharmaceutically-acceptable carriers include: (1) sugars, such as lactose, glucose and sucrose; (2) starches, such as corn starch and potato starch; (3) cellulose, and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; (4) powdered tragacanth; (5) malt; (6) gelatin; (7) talc; (8) excipients, such as cocoa butter and suppository waxes; (9) oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; (10) glycols, such as propylene glycol; (11) polyols, such as glycerin, sorbitol, mannitol and polyethylene glycol; (12) esters, such as ethyl oleate and ethyl laurate; (13) agar; (14) buffering agents, such as magnesium hydroxide and aluminum hydrox
  • Formulations of the present invention may be based in part on liposomes.
  • Liposomes consist of a phospholipid bilayer which forms a shell around an aqueous core.
  • Formulations of the present invention may be based in part on polymeric microparticles.
  • Microspheres formed of polymers or proteins are also well known to those skilled in the art, and can be tailored for passage through the gastrointestinal tract, as described in U.S. Pat. Nos. 4,906,474, 4,925,673, and 3,625,214, for example.
  • Biqerodible microspheres can be prepared using any of the methods developed for making microspheres for drug delivery, as described, for example, by Mathiowitz et al., J. Appl. Polymer Sci. 35, 755-774(1988), and P.
  • Coacervation/phase separation techniques have been used to encapsulate both solid and liquid core materials with various polymer coatings.
  • Simple coacervation employs a single colloid (e.g. gelatin in water) and involves the removal of the associated water from around the dispersed colloid by agents with a higher affinity for water, such as alcohols and salts.
  • Complex coacervation employs more than one colloid, and the separation proceeds mainly by charge neutralization of the colloids carrying opposite charges rather than by dehydration.
  • Coacervation may also be induced using nonaqueous vehicles, as described in Nakano et al., Int. J. Pharm, 4, 29-298(1980), for example.
  • Hydrogel microspheres made of gel-type polymers such as alginate or polyphosphazenes or other dicarboxylic polymers can be prepared by dissolving the polymer in an aqueous solution, suspending the material to be incorporated into the mixture, and extruding the polymer mixture through a microdroplet forming device, equipped with a nitrogen gas jet.
  • microspheres fall into a slowly stirring, ionic hardening bath, as illustrated, for example, by Salib, et al., Pharmazeutician Industrie 40- 11 A, 1230(1978), the teachings of which are incorporated herein.
  • the advantage of this system is the ability to further modify the surface ofthe microspheres by coating them with polycationic polymers (such as polylysine) after fabrication, as described, for example, by Lim et al, J. Pharm Sci. 70, 351-354(1981).
  • the microsphere particle size depends upon the extruder size as well as the polymer and gas flow rates.
  • polymers that can be used include polyamides, polycarbonates, polyalkylenes and derivatives thereof including, polyalkylene glycols, polyalkylene oxides, polyalkylene terepthalates, polymers of acrylic and methacrylic esters, including poly(methyl methacrylate), poly(ethyl methacrylate), poly(butylmethacrylate), poly(isobutyl methacrylate), poly(hexylmethacrylate), poly(isodecyl methacrylate), poly(lauryl methacrylate), poly(phenyl methacrylate), poly(methyl acrylate), poly(isopropyl acrylate), poly(isobutyl acrylate), and poly(octadecyl acrylate), polyvinyl polymers including polyvinyl alcohols, polyvinyl ethers, polyvinyl esters, polyvinyl halides, poly(vinyl acetate), and polyvinylpyrrolidone, polygly
  • biodegradable polymers include synthetic polymers such as polymers of lactic acid and glycolic acid, polyanhydrides, poly(ortho)esters, polyurethanes, poly(butic acid), poly(valeric acid), and poly(lactide-cocaprolactone), and natural polymers such as alginate and other polysaccharides including dextran and cellulose, collagen, chemical derivatives thereof (substitutions, additions of chemical groups, for example, alkyl, alkylene, hydroxylations, oxidations, and other modifications routinely made by those skilled in the art), albumin and other hydrophihc proteins, zein and other prolamines and hydrophobic proteins, copolymers and mixtures thereof.
  • synthetic polymers such as polymers of lactic acid and glycolic acid, polyanhydrides, poly(ortho)esters, polyurethanes, poly(butic acid), poly(valeric acid), and poly(lactide-cocaprolactone
  • natural polymers such as alginate and other polys
  • Bioadhesive polymers of particular interest include bioerodible hydrogels described by H. S. Sawhney, C. P. Pathak and J. A.
  • polyhyaluronic acids casein, gelatin, glutin, polyanhydrides, polyacrylic acid, alginate, chitosan, poly(methyl methacrylates), poly(ethyl methacrylates), poly(butylmethacrylate), poly(isobutyl methacrylate), poly(hexylmethacrylate), poly(isodecyl methacrylate), poly(lauryl methacrylate), poly(phenyl methacrylate), poly(methyl acrylate), poly(isopropyl acrylate), poly(isobutyl acrylate), and poly(octadecyl acrylate).
  • a diluent used in a composition of the present invention can be one or more compounds which are capable of densifying the active principle to give the desired mass.
  • the preferred diluents are mineral phosphates such as calcium phosphates; sugars such as hydrated or anhydrous lactose, or mannitol; and cellulose or cellulose derivatives, for example microcrystalline cellulose, starch, corn starch or pregelatinized starch.
  • Very particularly preferred diluents are lactose monohydrate, mannitol, microcrystalline cellulose and corn starch, used by themselves or in a mixtare, for example a mixtare of lactose monohydrate and corn starch or a mixtare of lactose monohydrate, corn starch and microcrystalline cellulose.
  • a binder employed in a composition of the present invention can be one or more compounds which are capable of densifying a compound of formula (I), converting it to coarser and denser particles with better flow properties.
  • the preferred binders are alginic acid or sodium alginate; cellulose and cellulose derivatives such as sodium carboxymethyl cellulose, ethyl cellulose, hydroxyethyl cellulose, hydroxypropyl cellulose, hydroxypropyl methyl cellulose or methyl cellulose, gelatin; acrylic acid polymers; and povidone, for example povidone K-30; hydroxypropyl methyl cellulose and povidone K-30 are very particularly preferred binders.
  • a disintegrating agent employed in a composition of the present invention can be one or more compounds which facilitate the disintegration of the prepared formulation when it is placed in an aqueous medium.
  • the preferred disintegrating agents are cellulose or cellulose derivatives such as sodium carboxymethyl cellulose, crosslinked sodium carboxymethyl cellulose, micro-crystalline cellulose, cellulose powder, crospovidone; pregelatinized starch, sodium starch glyconate, sodium carboxymethyl starch, or starch. Crospovidone, crosslinked sodium carboxymethyl cellulose and sodium carboxymethyl starch are preferred disintegrating agents.
  • An antiadhesive employed in a composition of the present invention can be one or more compounds which are capable of reducing the sticky character ofthe formulation, for example of preventing adhesion to metal surfaces.
  • the preferred antiadhesives are compounds containing silicon, for example silica or talcum.
  • a flow promoter employed in a composition of the present invention can be one or more compounds which are capable of facilitating the flow ofthe prepared formulation.
  • the preferred flow promoters are compounds containing silicon, for example anhydrous colloidal silica or precipitated silica.
  • a lubricant employed in a composition of the present invention can be one or more compounds which are capable of preventing the problems associated with the preparation of dry forms, such as the sticking and/or seizing problems which occur in the machines during compression or filling.
  • the preferred lubricants are fatty acids or fatty acid derivatives such as calcium stearate, glyceryl monostearate, glyceryl palmitostearate, magnesium stearate, sodium laurylsulfate, sodium stearylfumarate, zinc stearate or stearic acid; hydrogenated vegetable oils, for example hydrogenated castor oil; polyalkylene glycols or polyethylene glycol; sodium benzoate; or talcum.
  • Magnesium stearate or sodium stearylfumarate is preferred according to the present invention.
  • a color employed in a formulation of the present invention can be one or more compounds which are capable of imparting the desired color to the prepared formulation. ⁇ ie addition of a color can serve for example to differentiate between formulations containing different doses of active principle.
  • the preferred colors are iron oxides.
  • certain embodiments of the present compounds may contain a basic functional group, such as amino or alkylamino, and are, thus, capable of forming pharmaceutically-acceptable salts with pharmaceutically-acceptable acids.
  • pharmaceutically-acceptable salts refers to the relatively non-toxic, inorganic and organic acid addition salts of compounds of the present invention. These salts can be prepared in situ in the administration vehicle or the dosage form manufacturing process, or by separately reacting a purified compound of the invention in its free base form with a suitable organic or inorganic acid, and isolating the salt thus formed during subsequent purification.
  • Representative salts include the hydrobromide, hydrochloride, sulfate, bisulfate, phosphate, nitrate, acetate, valerate, oleate, palrnitate, stearate, laurate, benzoate, lactate, phosphate, tosylate, citrate, maleate, fumarate, succinate, tartrate, napthylate, mesylate, glucoheptonate, lactobionate, and laurylsulphonate salts and the like.
  • sulfate bisulfate
  • phosphate nitrate
  • acetate valerate
  • oleate palrnitate
  • stearate laurate
  • benzoate lactate, phosphate, tosylate, citrate, maleate, fumarate, succinate, tartrate, napthylate, mesylate, glucoheptonate, lactobionate, and laurylsulphonate salts and the like.
  • the pharmaceutically acceptable salts of the subject compounds include the conventional nontoxic salts or quaternary ammonium salts of the compounds, e.g., from non-toxic organic or inorganic acids.
  • such conventional nontoxic salts include those derived from inorganic acids such as hydrochloride, hydrobromic, sulfuric, sulfamic, phosphoric, nitric, and the like; and the salts prepared from organic acids such as acetic, propionic, succinic, glycolic, stearic, lactic, malic, tartaric, citric, ascorbic, palmitic, maleic, hydroxymaleic, phenylacetic, glutamic, benzoic, salicyclic, sulfanilic, 2- acetoxybenzoic, fumaric, toluenesulfonic, methanesulfonic, ethane disulfonic, oxalic, isothionic, and the like.
  • the compounds of the present invention may contain one or more acidic functional groups and, thus, are capable of forming pharmaceutically-acceptable salts with pharmaceutically-acceptable bases.
  • pharmaceutically-acceptable salts in these instances refers to the relatively non-toxic, inorganic and organic base addition salts of compounds of the present invention. These salts can likewise be prepared in situ in the administration vehicle or the dosage form manufacturing process, or by separately reacting the purified compound in its free acid form with a suitable base, such as the hydroxide, carbonate or bicarbonate of a pharmaceutically-acceptable metal cation, with ammonia, or with a pharmaceutically-acceptable organic primary, secondary or tertiary amine.
  • a suitable base such as the hydroxide, carbonate or bicarbonate of a pharmaceutically-acceptable metal cation, with ammonia, or with a pharmaceutically-acceptable organic primary, secondary or tertiary amine.
  • Representative alkali or alkaline earth salts include the lithium, sodium, potassium, calcium, magnesium, and aluminum salts and the like.
  • Representative organic amines useful for the formation of base addition salts include ethylamine, diethylamine, ethylenediamine, ethanolamine, diethanolamine, piperazine and the like. (See, for example, Berge et al., supra)
  • wetting agents such as sodium lauryl sulfate and magnesium stearate, as well as coloring agents, release agents, coating agents, sweetening, flavoring and perfuming agents, preservatives and antioxidants can also be present in the compositions.
  • antioxidants examples include: (1) water soluble antioxidants, such as ascorbic acid, cysteine hydrochloride, sodium bisulfate, sodium metabisulfite, sodium sulfite and the like; (2) oil-soluble antioxidants, such as ascorbyl palmitate, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), lecithin, propyl gallate, alpha-tocopherol, and the like; and (3) metal chelating agents, such as citric acid, ethylenediamine tetraacetic acid (EDTA), sorbitol, tartaric acid, phosphoric acid, and the like.
  • water soluble antioxidants such as ascorbic acid, cysteine hydrochloride, sodium bisulfate, sodium metabisulfite, sodium sulfite and the like
  • oil-soluble antioxidants such as ascorbyl palmitate, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), le
  • Formulations of the present invention include those suitable for oral, nasal, topical (including buccal and sublingual), rectal, vaginal and/or parenteral administration.
  • the fonnulations may conveniently be presented in unit dosage form and may be prepared by any methods well known in the art of pharmacy.
  • the amount of active ingredient which can be combined with a carrier material to produce a single dosage form will vary depending upon the host being treated, the particular mode of administration.
  • the amount of active ingredient which can be combined with a carrier material to produce a single dosage form will generally be that amount of the compound which produces a therapeutic effect.
  • a formulation of the present invention comprises an excipient selected from the group consisting of cyclodextrins, liposomes, micelle fonning agents, e.g., bile acids, and polymeric carriers, e.g., polyesters and polyanhydrides; and a compound of the present invention.
  • an aforementioned formulation renders orally bioavailable a compound ofthe present invention.
  • Methods of preparing these formulations or compositions include the step of bringing into association a compound of the present invention with the carrier and, optionally, one or more accessory ingredients.
  • the formulations are prepared by unifonnly and intimately bringing into association a compound of the present invention with liquid carriers, or finely divided solid carriers, or both, and then, if necessary, shaping the product.
  • Formulations of the invention suitable for oral administration may be in the form of capsules, cachets, pills, tablets, lozenges (using a flavored basis, usually sucrose and acacia or tragacanth), powders, granules, or as a solution or a suspension in an aqueous or non- aqueous liquid, or as an oil-in-water or water-in-oil liquid emulsion, or as an elixir or syrup, or as pastilles (using an inert base, such as gelatin and glycerin, or sucrose and acacia) and/or as mouth washes and the like, each containing a predetermined amount of a compound of the present invention as an active ingredient.
  • a compound of the present invention may also be administered as a bolus, electuary or paste.
  • the active ingredient is mixed with one or more pharmaceutically-acceptable carriers, such as sodium citrate or dicalcium phosphate, and/or any of the following: (1) fillers or extenders, such as starches, lactose, sucrose, glucose, mannitol, and/or silicic acid; (2) binders, such as, for example, carboxymethylcellulose, alginates, gelatin, polyvinyl pyrrolidone, sucrose and/or acacia; (3) humectants, such as glycerol; (4) disintegrating agents, such as agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates, and sodium carbonate; (5) solution retarding agents, such as paraffin; (6) absorption accelerators, such as quaternary ammonium compounds; (7) wetting agents, such as, for example,
  • compositions may also comprise buffering agents.
  • Solid compositions of a similar type may also be employed as fillers in soft and hard-shelled gelatin capsules using such excipients as lactose or milk sugars, as well as high molecular weight polyethylene glycols and the like.
  • a tablet may be made by compression or molding, optionally with one or more accessory ingredients.
  • Compressed tablets may be prepared using binder (for example, gelatin or hydroxypropylmethyl cellulose), lubricant, inert diluent, preservative, disintegrant
  • Molded tablets may be made by molding in a suitable machine a mixtare ofthe powdered compound moistened with an inert liquid diluent.
  • the tablets, and other solid dosage forms of the pharmaceutical compositions of the present invention may optionally be scored or prepared with coatings and shells, such as enteric coatings and other coatings well known in the pharmaceutical-formulating art. They may also be formulated so as to provide slow or controlled release of the active ingredient therein using, for example, hydroxypropylmethyl cellulose in varying proportions to provide the desired release profile, other polymer matrices, liposomes and/or microspheres. They may be formulated for rapid release, e.g., freeze-dried.
  • compositions may be sterilized by, for example, filtration through a bacteria-retaining filter, or by incorporating sterilizing agents in the form of sterile solid compositions which can be dissolved in sterile water, or some other sterile injectable medium immediately before use.
  • These compositions may also optionally contain opacifying agents and may be of a composition that they release the active ingredient(s) only, or preferentially, in a certain portion of the gastrointestinal tract, optionally, in a delayed manner.
  • embedding compositions which can be used include polymeric substances and waxes.
  • the active ingredient can also be in micro-encapsulated form, if appropriate, with one or more of the above-described excipients.
  • Liquid dosage forms for oral administration of the compounds of the invention include pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups and elixirs.
  • the liquid dosage forms may contain inert diluents commonly used in the art, such as, for example, water or other solvents, solubilizing agents and emulsifiers, such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, oils (in particular, cottonseed, groundnut, com, germ, olive, castor and sesame oils), glycerol, tetrahydrofuryl alcohol, polyethylene glycols and fatty acid esters of sorbitan, and mixtures thereof.
  • inert diluents commonly used in the art, such as, for example, water or other solvents, solubilizing agents
  • the oral compositions can also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, coloring, perfuming and preservative agents.
  • adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, coloring, perfuming and preservative agents.
  • Suspensions in addition to the active compounds, may contain suspending agents as, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar-agar and tragacanth, and mixtures thereof.
  • suspending agents as, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar-agar and tragacanth, and mixtures thereof.
  • Formulations of the pharmaceutical compositions of the invention for rectal or vaginal administration may be presented as a suppository, which may be prepared by mixing one or more compounds of the invention with one or more suitable nonirritating excipients or carriers comprising, for example, cocoa butter, polyethylene glycol, a suppository wax or a salicylate, and which is solid at room temperature, but liquid at body temperature and, therefore, will melt in the rectum or vaginal cavity and release the active compound.
  • Formulations of the present invention which are suitable for vaginal administration also include pessaries, tampons, creams, gels, pastes, foams or spray formulations containing such carriers as are known in the art to be appropriate.
  • Dosage forms for the topical or transdermal administration of a compound of this invention include powders, sprays, ointments, pastes, creams, lotions, gels, solutions, patches and inhalants.
  • the active compound may be mixed under sterile conditions with a pharmaceutically-acceptable carrier, and with any preservatives, buffers, or propellants which may be required.
  • the ointments, pastes, creams and gels may contain, in addition to an active compound of this invention, excipients, such as animal and vegetable fats, oils, waxes, paraffins, starch, tragacanth, cellulose derivatives, polyethylene glycols, silicones, bentonites, silicic acid, talc and zinc oxide, or mixtures thereof.
  • excipients such as animal and vegetable fats, oils, waxes, paraffins, starch, tragacanth, cellulose derivatives, polyethylene glycols, silicones, bentonites, silicic acid, talc and zinc oxide, or mixtures thereof.
  • Powders and sprays can contain, in addition to a compound of this invention, excipients such as lactose, talc, silicic acid, aluminum hydroxide, calcium silicates and polyamide powder, or mixtures of these substances.
  • Sprays can additionally contain customary propellants, such as chlorofluorohydrocarbons and volatile unsubstitated hydrocarbons, such as butane and propane.
  • Transdermal patches have the added advantage of providing controlled delivery of a compound of the present invention to the body.
  • dosage forms can be made by dissolving or dispersing the compound in the proper medium.
  • Absorption enhancers can also be used to increase the flux of the compound across the skin. The rate of such flux can be controlled by either providing a rate controlling membrane or dispersing the compound in a polymer matrix or gel.
  • Ophthalmic formulations, eye ointments, powders, solutions and the like, are also contemplated as being within the scope of this invention.
  • compositions of this invention suitable for parenteral administration comprise one or more compounds of the invention in combination with one or more pharmaceutically-acceptable sterile isotonic aqueous or nonaqueous solutions, dispersions, suspensions or emulsions, or sterile powders which may be reconstituted into sterile injectable solutions or dispersions just prior to use, which may contain sugars, alcohols, antioxidants, buffers, bacteriostats, solutes which render the formulation isotonic with the blood ofthe intended recipient or suspending or thickening agents.
  • aqueous and nonaqueous carriers examples include water, ethanol, polyols (such as glycerol, propylene glycol, polyethylene glycol, and the like), and suitable mixtures thereof, vegetable oils, such as olive oil, and injectable organic esters, such as ethyl oleate.
  • polyols such as glycerol, propylene glycol, polyethylene glycol, and the like
  • vegetable oils such as olive oil
  • injectable organic esters such as ethyl oleate.
  • Proper fluidity can be maintained, for example, by the use of coating materials, such as lecithin, by the maintenance of the required particle size in the case of dispersions, and by the use of surfactants.
  • compositions may also contain adjuvants such as preservatives, wetting agents, emulsifying agents and dispersing agents. Prevention of the action of microorganisms upon the subject compounds may be ensured by the inclusion of various antibacterial and antifungal agents, for example, paraben, chlorobutanol, phenol sorbic acid, and the like. It may also be desirable to include isotonic agents, such as sugars, sodium chloride, and the like into the compositions. In addition, prolonged absorption of the injectable pharmaceutical form may be brought about by the inclusion of agents which delay absorption such as aluminum monostearate and gelatin. In some cases, in order to prolong the effect of a drug, it is desirable to slow the absorption of the drug from subcutaneous or intramuscular injection.
  • adjuvants such as preservatives, wetting agents, emulsifying agents and dispersing agents.
  • Injectable depot forms are made by forming microencapsule matrices of the subject compounds in biodegradable polymers such as polylactide-polyglycolide. Depending on the ratio of drug to polymer, and the nature ofthe particular polymer employed, the rate of drug release can be controlled. Examples of other biodegradable polymers include poly(orthoesters) and poly(anhydrides). Depot injectable formulations are also prepared by entrapping the drug in liposomes or microemulsions which are compatible with body tissue.
  • the compounds of the present invention When the compounds of the present invention are administered as pharmaceuticals, to humans and animals, they can be given per se or as a pharmaceutical composition containing, for example, 0.1 to 99.5% (more preferably, 0.5 to 90%) of active ingredient in combination with a pharmaceutically acceptable carrier.
  • the preparations of the present invention may be given orally, parenterally, topically, or rectally. They are of course given in forms suitable for each administration route. For example, they are administered in tablets or capsule form, by injection, inhalation, eye lotion, ointment, suppository, etc. administration by injection, infusion or inhalation; topical by lotion or ointment; and rectal by suppositories. Oral administrations are preferred.
  • parenteral administration and “administered parenterally” as used herein means modes of administration other than enteral and topical administration, usually by injection, and includes, without limitation, intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcuticular, intraarticulare, subcapsular, subarachnoid, intraspinal and intrasternal injection and infusion.
  • systemic administration means the administration of a compound, drug or other material other than directly into the central nervous system, such that it enters the patient's system and, thus, is subject to metabolism and other like processes, for example, subcutaneous administration.
  • These compounds may be administered to humans and other animals for therapy by any suitable route of administration, including orally, nasally, as by, for example, a spray, rectally, intravaginally, parenterally, intracisternally and topically, as by powders, ointments or drops, including buccally and sublingually.
  • the compounds of the present invention which may be used in a suitable hydrated form, and/or the pharmaceutical compositions of the present invention, are formulated into pharmaceutically-acceptable dosage forms by conventional methods known to those of skill in the art.
  • Actual dosage levels of the active ingredients in the pharmaceutical compositions of this invention may be varied so as to obtain an amount of the active ingredient which is effective to achieve the desired therapeutic response for a particular patient, composition, and mode of administration, without being toxic to the patient.
  • the selected dosage level will depend upon a variety of factors including the activity of the particular compound of the present invention employed, or the ester, salt or amide thereof, the route of administration, the time of administration, the rate of excretion or metabolism of the particular compound being employed, the duration of the treatment, other drugs, compounds and/or materials used in combination with the particular compound employed, the age, sex, weight, condition, general health and prior medical history of the patient being treated, and like factors well known in the medical arts.
  • a physician or veterinarian having ordinary skill in the art can readily determine and prescribe the effective amount of the pharmaceutical composition required.
  • the physician or veterinarian could start doses of the compounds of the invention employed in the pharmaceutical composition at levels lower than that required in order to achieve the desired therapeutic effect and gradually increase the dosage until the desired effect is achieved.
  • a suitable daily dose of a compound of the invention will be that amount ofthe compound which is the lowest dose effective to produce a therapeutic effect.
  • Such an effective dose will generally depend upon the factors described above.
  • intravenous, intracerebroventricular and subcutaneous doses of the compounds of this invention for a patient, when used for the indicated analgesic effects will range from about 0.0001 to about 100 mg per kilogram of body weight per day.
  • the effective daily dose of the active compound may be administered as two, three, four, five, six or more sub-doses administered separately at appropriate intervals throughout the day, optionally, in unit dosage forms.
  • composition While it is possible for a compound of the present invention to be administered alone, it is preferable to administer the compound as a pharmaceutical formulation (composition).
  • the present invention provides pharmaceutically acceptable compositions which comprise a therapeutically-effective amount of one or more of the subject compounds, as described above, formulated together with one or more pharmaceutically acceptable carriers (additives) and/or diluents.
  • compositions of the present invention may be specially formulated for administration in solid or liquid form, including those adapted for the following: (1) oral administration, for example, drenches (aqueous or non-aqueous solutions or suspensions), tablets, boluses, powders, granules, pastes for application to the tongue; (2) parenteral administration, for example, by subcutaneous, intramuscular or intravenous injection as, for example, a sterile solution or suspension; (3) topical application, for example, as a cream, ointment or spray applied to the skin, lungs, or oral cavity; or (4) intravaginally or intravectally, for example, as a pessary, cream or foam; (5) sublingually; (6) ocularly; (7) transdermally; or (8) nasally.
  • oral administration for example, drenches (aqueous or non-aqueous solutions or suspensions), tablets, boluses, powders, granules, pastes for application to the tongue
  • parenteral administration for example
  • the compounds according to the invention may be formulated for administration in any convenient way for use in human or veterinary medicine, by analogy with other pharmaceuticals.
  • treatment is intended to encompass also prophylaxis, therapy and cure.
  • the patient receiving this treatment is any animal in need, including primates, in particular humans, and other mammals such as equines, cattle, swine and sheep; and poultry and pets in general.
  • the compound of the invention can be administered as such or in admixtures with pharmaceutically acceptable carriers and can also be administered in conjunction with antimicrobial agents such as penicillins, cephalosporins, aminoglycosides and glycopeptides.
  • Conjunctive therapy thus includes sequential, simultaneous and separate administration of the active compound in a way that the therapeutical effects of the first administered one is not entirely disappeared when the subsequent is administered.
  • the addition of the active compound of the invention to animal feed is preferably accomplished by preparing an appropriate feed premix containing the active compound in an effective amount and inco ⁇ orating the premix into the complete ration.
  • an intermediate concentrate or feed supplement containing the active ingredient can be blended into the feed.
  • feed premixes and complete rations can be prepared and administered are described in reference books (such as "Applied Animal Nutrition", W.H. Freedman and CO., San Francisco, U.S.A., 1969 or “Livestock Feeds and Feeding" O and B books, Corvallis, Ore., U.S.A., 1977).
  • a combinatorial library for the pu ⁇ oses of the present invention is a mixture of chemically related compounds which may be screened together for a desired property; said libraries may be in solution or covalently linked to a solid support.
  • the preparation of many related compounds in a single reaction greatly reduces and simplifies the number of screening processes which need to be carried out. Screening for the appropriate biological, pharmaceutical, agrochemical or physical property may be done by conventional methods.
  • the substrate aryl groups used in a combinatorial approach can be diverse in terms of the core aryl moiety, e.g., a variegation in terms of the ring structare, and/or can be varied with respect to the other substituents.
  • a variety of libraries on the order of about 16 to 1,000,000 or more diversomers can be synthesized and screened for a particular activity or property.
  • a library of substituted diversomers can be synthesized using the subject reactions adapted to the techniques described in the Still et al. PCT publication WO 94/08051, e.g., being linked to a polymer bead by a hydrolyzable or photolyzable group, e.g., located at one of the positions of substrate.
  • the library is synthesized on a set of beads, each bead including a set of tags identifying the particular diversomer on that bead.
  • the beads can be dispersed on the surface of a permeable membrane, and the diversomers released from the beads by lysis of the bead linker.
  • the diversomer from each bead will diffuse across the membrane to an assay zone, where it will interact with an enzyme assay.
  • MS mass spectrometry
  • a compound selected from a combinatorial library can be irradiated in a MALDI step in order to release the diversomer from the matrix, and ionize the diversomer for MS analysis.
  • the libraries of the subject method can take the multipin library format.
  • Geysen and co-workers (Geysen et al. (1984) PNAS 81:3998-4002) introduced a method for generating compound libraries by a parallel synthesis on polyacrylic acid-grated polyethylene pins arrayed in the microtifre plate format.
  • the Geysen technique can be used to synthesize and screen thousands of compounds per week using the multipin method, and the tethered compounds may be reused in many assays.
  • Appropriate linker moieties can also been appended to the pins so that the compounds may be cleaved from the supports after synthesis for assessment of purity and further evaluation (c.f, Bray et al. (1990) Tetrahedron Lett 31:5811-5814; Valerio et al. (1991) Anal Biochem 197:168-177; Bray et al. (1991) Tetrahedron Lett 32:6163-6166).
  • a variegated library of compounds can be provided on a set of beads utilizing the strategy of divide-couple-recombine (see, e.g., Houghten (1985) PNAS 82:5131-5135; and U.S. Patents 4,631,211; 5,440,016; 5,480,971).
  • the beads are divided into separate groups equal to the number of different substitaents to be added at a particular position in the library, the different substitaents coupled in separate reactions, and the beads recombined into one pool for the next iteration.
  • the divide-couple-recombine strategy can be carried out using an analogous approach to the so-called "tea bag” method first developed by Houghten, where compound synthesis occurs on resin sealed inside porous polypropylene bags (Houghten et al. (1986) PNAS 82:5131-5135). Substituents are coupled to the compound- bearing resins by placing the bags in appropriate reaction solutions, while all common steps such as resin washing and deprotection are performed simultaneously in one reaction vessel. At the end ofthe synthesis, each bag contains a single compound. D) Combinatorial Libraries by Light-Directed, Spatially Addressable Parallel Chemical Synthesis
  • a scheme of combinatorial synthesis in which the identity of a compound is given by its locations on a synthesis substrate is termed a spatially-addressable synthesis.
  • the combinatorial process is carried out by controlling the addition of a chemical reagent to specific locations on a solid support (Dower et al. (1991) Annu Rep Med Chem 26:271-280; Fodor, S.P.A. (1991) Science 251:767; Pirrung et al. (1992) U.S. Patent No. 5,143,854; Jacobs et al. (1994) Trends Biotechnol 12:19-26).
  • the spatial resolution of photolithography affords miniaturization. This technique can be carried out through the use protection/deprotection reactions with photolabile protecting groups. The key points of this technology are illustrated in Gallop et al. (1994) J Med Chem
  • a synthesis substrate is prepared for coupling through the covalent attachment of photolabile nitroveratryloxycarbonyl (NVOC) protected amino linkers or other photolabile linkers.
  • Light is used to selectively activate a specified region of the synthesis support for coupling. Removal of the photolabile protecting groups by light (deprotection) results in activation of selected areas. After activation, the first of a set of amino acid analogs, each bearing a photolabile protecting group on the amino terminus, is exposed to the entire surface. Coupling only occurs in regions that were addressed by light in the preceding step. The reaction is stopped, the plates washed, and the substrate is again illuminated through a second mask, activating a different region for reaction with a second protected building block.
  • NVOC photolabile nitroveratryloxycarbonyl
  • the pattern of masks and the sequence of reactants define the products and their locations. Since this process utilizes photolithography techniques, the number of compounds that can be synthesized is limited only by the number of synthesis sites that can be addressed with appropriate resolution. The position of each compound is precisely known; hence, its interactions with other molecules can be directly assessed.
  • the subject method utilizes a compound library provided with an encoded tagging system.
  • a recent improvement in the identification of active compounds from combinatorial libraries employs chemical indexing systems using tags that uniquely encode the reaction steps a given bead has undergone and, by inference, the structure it carries.
  • this approach mimics phage display libraries, where activity derives from expressed peptides, but the structures of the active peptides are deduced from the corresponding genomic DNA sequence.
  • the first encoding of synthetic combinatorial libraries employed DNA as the code.
  • a variety of other forms of encoding have been reported, including encoding with sequenceable bio-oligomers (e.g., oligonucleotides and peptides), and binary encoding with additional non-sequenceable tags.
  • a combinatorial library of nominally 7 ⁇ ( 823,543) peptides composed of all combinations of Arg, Gin, Phe, Lys, Val, D-Val and Thr (three-letter amino acid code), each of which was encoded by a specific dinucleotide (TA, TC, CT, AT, TT, CA and AC, respectively), was prepared by a series of alternating rounds of peptide and oligonucleotide synthesis on solid support.
  • the amine linking functionality on the bead was specifically differentiated toward peptide or oligonucleotide synthesis by simultaneously preincubating the beads with reagents that generate protected OH groups for oligonucleotide synthesis and protected H2 groups for peptide synthesis (here, in a ratio of 1:20).
  • the tags each consisted of 69-mers, 14 units of which carried the code.
  • the bead-bound library was incubated with a fiuorescently labeled antibody, and beads containing bound antibody that fluoresced strongly were harvested by fluorescence- activated cell sorting (FACS).
  • FACS fluorescence- activated cell sorting
  • compound libraries can be derived for use in the subject method, where the oligonucleotide sequence ofthe tag identifies the sequential combinatorial reactions that a particular bead underwent, and therefore provides the identity of the compound on the bead.
  • oligonucleotide tags permits extremelyly sensitive tag analysis. Even so, the method requires careful choice of orthogonal sets of protecting groups required for alternating co-synthesis of the tag and the library member. Furthermore, the chemical lability of the tag, particularly the phosphate and sugar anomeric linkages, may limit the choice of reagents and conditions that can be employed for the synthesis of non-oligomeric libraries. In preferred embodiments, the libraries employ linkers permitting selective detachment ofthe test compound library member for assay.
  • Peptides have also been employed as tagging molecules for combinatorial libraries.
  • Two exemplary approaches are described in the art, both of which employ branched linkers to solid phase upon which coding and ligand strands are alternately elaborated.
  • orthogonality in synthesis is achieved by employing acid-labile protection for the coding strand and base-labile protection for the compound strand.
  • branched linkers are employed so that the coding unit and the test compound can both be attached to the same functional group on the resin.
  • a cleavable linker can be placed between the branch point and the bead so that cleavage releases a molecule containing both code and the compound (Ptek et al. (1991) Tetrahedron Lett 32:3891-3894).
  • the cleavable linker can be placed so that the test compound can be selectively separated from the bead, leaving the code behind. This last construct is particularly valuable because it permits screening of the test compound without potential interference of the coding groups. Examples in the art of independent cleavage and sequencing of peptide library members and their corresponding tags has confirmed that the tags can accurately predict the peptide structure. 2) Non-sequenceable Tagging: Binary Encoding
  • An alternative form of encoding the test compound library employs a set of non-sequencable electrophone tagging molecules that are used as a binary code (Ohlmeyer et al. (1993) PNAS 90:10922-10926).
  • Exemplary tags are haloaromatic alkyl ethers that are detectable as their trirnethylsilyl ethers at less than femtomolar levels by electron capture gas chromatography (ECGC). Variations in the length of the alkyl chain, as well as the nature and position of the aromatic halide substitaents, permit the synthesis of at least 40 such tags, which in principle can encode 2 ⁇ 0 (e.g., upwards of 10 ⁇ ) different molecules.
  • Both libraries were constructed using an orthogonal attachment strategy in which the library member was linked to the solid support by a photolabile linker and the tags were attached through a linker cleavable only by vigorous oxidation. Because the library members can be repetitively partially photoeluted from the solid support, library members can be utilized in multiple assays. Successive photoelution also permits a very high throughput iterative screening strategy: first, multiple beads are placed in 96-well microtiter plates; second, compounds are partially detached and transferred to assay plates; third, a metal binding assay identifies the active wells; fourth, the corresponding beads are rearrayed singly into new microtiter plates; fifth, single active compounds are identified; and sixth, the structures are decoded.
  • Rotenone 25 g, 0.063 mol was placed in a 100 mL flask equipped with a stir bar.
  • the white solid was dissolved in 500 mL of methanol.
  • trimethyl orthoformate 8.31 mL, 0.076 mol
  • p-toluenesulfonic acid catalytic amount was added.
  • the solution was refluxed for 16 hrs, with additional 0.1 g of catalyst added after 8 hrs.
  • the solution was then allowed to cool to room temperature. Neutralization of the reaction was accomplished by addition of potassium carbonate.
  • the solution was then vacuumed down to residue.
  • Rotenone methyl enol ether (1.51 g, 3.69 mmol) was placed in a 250 mL flask equipped with a stirrer under nitrogen. The yellow solid was dissolved in 9-BBN (0.5 M solution in THF) (29.6 mL, 0.0148 mol). The mixtare was refluxed for 5 hrs. After cooling to room temperatare 6 mL of ethanol were added, followed by 2 mL of 6 N NaOH, and finally 4 mL of 30% H 2 0 2 . Hie additions were all performed dropwise resulting in a cloudy mixture which was then heated at 50 C for 1 hr.
  • 9-BBN 0.5 M solution in THF
  • the solution was cooled, extracted with ether, washed with a solution of saturated sodium carbonate, and dried over sodium sulfate. The solution was then vacuumed down to residue. The residue was passed through a silca column using 1% methanol / methylene chloride yielding 0.660 g, 42% yield.
  • the hydroxymethyl rotenone derivative (0.568 g, 1.33 mmol) was placed in a 100 mL flask equipped with a stirrer under nitrogen. The flask containing the yellow solid was placed in an ice-bath. The solid was dissolved in 20 mL of methylene chloride, followed by addition of triethylamine (0.185 mL, 2.66 mmol). Finally, p-toluenesulfonyl chloride (0.508 g, 2.66 mmol) was added to the reaction mixture. The solution was allowed to stir at room temperatare for 4 hrs. The solution was then vacuumed down to residue.
  • the tosylated rotenone derivative (0.350 g, 0.60 mmol) was placed in a 100 mL flask equipped with a stirrer under nitrogen. The solid was dissolved in 5 mL of DMF, followed by addition of potassium carbonate (0.10 g, 0.72 mmol). Finally, the ethyl ester of picolineamine mono-acetic acid (PAMA) was added (0.138 g, 0.664 mmol) was added to the reaction mixture. The solution was heated at 105 C for 4 hrs. The solution was then vacuumed down to residue. The residue was passed through a HPLC silca gel column using 0-10% methanol/methylene chloride as the solvents, yielding 0.066 g, 18% yield.
  • PAMA picolineamine mono-acetic acid
  • Example 4 The procedure outlined in Example 4 was used with a mesylated rotenone derivative, yielding 0.006 g, 21% yield.
  • Example 4 The procedure outlined in Example 4 was used with a mesylated rotenone derivative and protected N,N'-bis(PMB-S-ethyl)glycine amide, yielding 0.010 g ofthe crude product.
  • Dipyridinemethylamine (DPMA) (0.0035 g, 0.0018 mmol) and o-Tos-Rotenol (0.01 g, 0.0018 mmol) were mixed in a 100 mL pressure tube in 2 mL of DMF under nitrogen.
  • Potassium carbonate (0.05 g) and triethylamine (0.3 mL) were added to the solution.
  • the mixtare was heated at 130° C for 3 hrs.
  • the reaction mixture was vacuumed down to residue.
  • the residue was purified through a pad of silica gel using methanol-methylene chloride to provide the product in 51 % yield.
  • Re(CO) 3 ( ⁇ 3 -(rotenol-dipyridinemethylamine)
  • the synthesis of the Re(I) tricarbonyl complex was accomplished by reacting [NEt 4 ] 2 [ReBr 3 (CO) 3 ] with the rotenol-DPMA in the ratio of 1:1.2 in 2.5 mL of methanol.
  • the reaction mixture was heated at 120 °C for 6 hours.
  • the reaction product was purified using a small silica column using 95% methylene chloride 5% methanol.
  • LCMS analysis using a C18 column with water and acetonitrile demonstrated multiple peaks, but near the retention time of the 99m Tc-labeled product demonstrated the molecular weight 864, which corresponds to the rhenium complex.
  • the method employed was a gradient 5-95% B, 1 mL/ minute for 30 minutes. The gradient ramped from 5-95 from 3-20 minutes. In challenge experiments the HPLC purified product demonstrated no degradation in either 10 mM Cysteine or Histidine in PBS pH 7.2 at 37° C for 18 hrs.
  • the complex was injected via the tail vein in 10% ethanol in saline (10 ⁇ Ci / 100 ⁇ l). Animals were sacrificed at 5, 15, 30, and 60 minutes p.i. The results are shown below in Table 1.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Optics & Photonics (AREA)
  • Chemical & Material Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Physics & Mathematics (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Epidemiology (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)

Abstract

Selon un aspect, l'invention concerne des dérivés de roténone à marquage au Tc-99m, qui sont utiles, par exemple, comme agents d'imagerie de flux sanguin myocardique. On décrit la synthèse, le radiomarquage et l'essai de complexes de roténone à marquage auTc-99m, en tant que sondes d'émission monophotonique, permettant d'améliorer la délimitation de perfusion myocardique. Les dérivés considérés ont des propriétés uniques. L'invention concerne également des dérivés de ce type qui ont un potentiel élevé d'absorption et de rétention dans le myocarde en fonction du flux. Selon un autre aspect, l'invention concerne des kits qui renferment un dérivé de roténone à marquage au Tc-99m.
PCT/US2003/008883 2002-04-08 2003-03-21 Derives de rotenone a marquage au technetium, et procedes d'utilisation correspondants WO2003086476A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
AU2003224747A AU2003224747A1 (en) 2002-04-08 2003-03-21 Technetium-labeled rotenone derivatives, and methods of use thereof

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US37066802P 2002-04-08 2002-04-08
US60/370,668 2002-04-08

Publications (1)

Publication Number Publication Date
WO2003086476A1 true WO2003086476A1 (fr) 2003-10-23

Family

ID=29250567

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2003/008883 WO2003086476A1 (fr) 2002-04-08 2003-03-21 Derives de rotenone a marquage au technetium, et procedes d'utilisation correspondants

Country Status (2)

Country Link
AU (1) AU2003224747A1 (fr)
WO (1) WO2003086476A1 (fr)

Cited By (20)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2005079391A3 (fr) * 2004-02-13 2006-02-16 Bristol Myers Squibb Pharma Co Agents de contraste pour l'imagerie de perfusion myocardique
EP1740225A2 (fr) * 2004-04-28 2007-01-10 Bristol-Myers Squibb Company Agents de contraste pour l'imagerie de perfusion myocardique
WO2007137135A2 (fr) * 2006-05-18 2007-11-29 The Regents Of The University Of California Analogues de roténone, méthode de préparation et d'utilisation
WO2008133730A2 (fr) * 2006-11-15 2008-11-06 The Regents Of The University Of California Analogues de roténone marqués
US7824659B2 (en) 2005-08-10 2010-11-02 Lantheus Medical Imaging, Inc. Methods of making radiolabeled tracers and precursors thereof
WO2010127054A3 (fr) * 2009-04-30 2011-01-06 Ge Healthcare Limited Imagerie d'une perfusion de tumeur et du métabolisme oxydatif à l'aide d'ace pet dynamique chez des patients atteints du cancer de la tête et du cou durant une radiothérapie
CN102336764A (zh) * 2011-06-21 2012-02-01 兰州大学 自旋标记鱼藤酮肟酯及其制备方法和用途
WO2013006958A1 (fr) 2011-07-13 2013-01-17 Nordion (Canada) Inc. Dérivés de roténone radio-marqués et leur utilisation en imagerie spect
US8551448B2 (en) 2006-05-18 2013-10-08 The Regents Of The University Of California Rotanone analogs: method of preparation and use
US8926945B2 (en) 2005-10-07 2015-01-06 Guerbet Compounds comprising a biological target recognizing part, coupled to a signal part capable of complexing gallium
US8936777B2 (en) 2010-02-08 2015-01-20 Lantheus Medical Imaging, Inc. Methods and apparatus for synthesizing imaging agents, and intermediates thereof
US8986650B2 (en) 2005-10-07 2015-03-24 Guerbet Complex folate-NOTA-Ga68
US9408927B2 (en) 2008-02-29 2016-08-09 Lantheus Medical Imaging, Inc. Contrast agents for applications including perfusion imaging
US9579408B2 (en) 2011-02-11 2017-02-28 Washington University PET/SPECT agents for applications in biomedical imaging
US9687571B2 (en) 2009-04-15 2017-06-27 Lantheus Medical Imaging, Inc. Stabilization of radiopharmaceutical compositions using ascorbic acid
US9713651B2 (en) 2012-08-10 2017-07-25 Lantheus Medical Imaging, Inc. Compositions, methods, and systems for the synthesis and use of imaging agents
US9850183B2 (en) 2014-06-27 2017-12-26 Reiley Pharmaceuticals, Inc. Conjugates derived from non-steroidal anti-inflammatory drugs and methods of use thereof in imaging
US10053478B2 (en) 2015-01-09 2018-08-21 Reiley Pharmaceuticals, Inc. COX-2-targeting, platinum-containing conjugates and their use in the treatment of tumors and cancers
US20210024555A1 (en) * 2018-03-22 2021-01-28 National Centre For Scientific Research "Demokritos" Tricarbonyl complexes of transition metals with benzo-heterocyclic derivatives of the cyclopentadienyl anion
WO2022272268A1 (fr) * 2021-06-23 2022-12-29 Yale University Composés pour l'imagerie moléculaire du renouvellement de collagène et méthodes les utilisant

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6051207A (en) * 1996-10-28 2000-04-18 Nycomed Imaging As Contrast agents

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6051207A (en) * 1996-10-28 2000-04-18 Nycomed Imaging As Contrast agents

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
ENAS J.D.: "(125I)Iodorotenone and (125I)iodorotenol", JOURNAL OF LABELLED COMPOUNDS AND RADIOPHARMACEUTICALS, SYMPOSIUM ABSTRACTS, vol. 37, 1995, pages 220 - 222, XP002973144 *

Cited By (58)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US9161997B2 (en) 2001-08-16 2015-10-20 David S. Casebier Contrast agents for myocardial perfusion imaging
JP2014218525A (ja) * 2004-02-13 2014-11-20 ブリストル−マイヤーズ・スクイブ・ファーマ・カンパニーＢｒｉｓｔｏｌ−Ｍｙｅｒｓ Ｓｑｕｉｂｂ Ｐｈａｒｍａ Ｃｏｍｐａｎｙ 心筋灌流イメージングのための造影剤
US10125106B2 (en) 2004-02-13 2018-11-13 Lantheus Medical Imaging, Inc. Contrast agents for myocardial perfusion imaging
AU2005214898B2 (en) * 2004-02-13 2011-01-06 Lantheus Medical Imaging, Inc. Contrast agents for myocardial perfusion imaging
US7344702B2 (en) 2004-02-13 2008-03-18 Bristol-Myers Squibb Pharma Company Contrast agents for myocardial perfusion imaging
WO2005079391A3 (fr) * 2004-02-13 2006-02-16 Bristol Myers Squibb Pharma Co Agents de contraste pour l'imagerie de perfusion myocardique
US9718786B2 (en) 2004-02-13 2017-08-01 Lantheus Medical Imaging, Inc. Contrast agents for myocardial perfusion imaging
JP2013047277A (ja) * 2004-02-13 2013-03-07 Bristol-Myers Squibb Pharma Co 心筋灌流イメージングのための造影剤
AU2005214898C1 (en) * 2004-02-13 2011-08-04 Lantheus Medical Imaging, Inc. Contrast agents for myocardial perfusion imaging
EP3385253B1 (fr) * 2004-02-13 2021-04-07 Lantheus Medical Imaging, Inc. Agents de contraste pour imagerie de perfusion du myocarde
KR101186449B1 (ko) 2004-02-13 2012-09-27 브리스톨-마이어스 스퀴브 파마 컴퍼니 심근 관류 영상화용 조영제
US8226929B2 (en) 2004-02-13 2012-07-24 Lantheus Medical Imaging, Inc. Contrast agents for myocardial perfusion imaging
KR101386851B1 (ko) 2004-02-13 2014-04-17 랜티우스 메디컬 이메징, 인크. 심근 관류 영상화용 조영제
RU2648358C2 (ru) * 2004-02-13 2018-03-23 Лантеус Медикал Имэджинг,Инк. Способ получения изображения кровоснабжения миокарда
JP2007526916A (ja) * 2004-02-13 2007-09-20 ブリストル−マイヤーズ・スクイブ・ファーマ・カンパニー 心筋灌流イメージングのための造影剤
NO344857B1 (no) * 2004-02-13 2020-06-02 Lantheus Medical Imaging Inc Kontrastmidler, forløperforbindelse for disse, preparater inneholdende slike, fremgangsmåte for syntese samt anvendelse av disse til avbilding
CN1925878B (zh) * 2004-02-13 2011-10-12 布里斯托尔-迈尔斯·斯奎布制药公司 供心肌灌注成像所用的对比剂
CN102274527A (zh) * 2004-02-13 2011-12-14 布里斯托尔-迈尔斯.斯奎布制药公司 供心肌灌注成像所用的对比剂
US10889550B2 (en) 2004-02-13 2021-01-12 Lantheus Medical Imaging, Inc. Contrast agents for myocardial perfusion imaging
EP1740225A2 (fr) * 2004-04-28 2007-01-10 Bristol-Myers Squibb Company Agents de contraste pour l'imagerie de perfusion myocardique
US8263042B2 (en) 2004-04-28 2012-09-11 Lantheus Medical Imaging, Inc. Contrast agents for myocardial perfusion imaging
CN101843908A (zh) * 2004-04-28 2010-09-29 布里斯托尔-迈尔斯斯奎布药品公司 心肌灌注成像的造影剂
EP1740225A4 (fr) * 2004-04-28 2009-08-19 Bristol Myers Squibb Co Agents de contraste pour l'imagerie de perfusion myocardique
US7485283B2 (en) 2004-04-28 2009-02-03 Lantheus Medical Imaging Contrast agents for myocardial perfusion imaging
EP2253333A3 (fr) * 2004-04-28 2013-05-08 Lantheus Medical Imaging, Inc. Agents de contraste pour l'imagerie de perfusion
AU2005238080B2 (en) * 2004-04-28 2011-02-10 Lantheus Medical Imaging, Inc. Contrast agents for myocardial perfusion imaging
US7824659B2 (en) 2005-08-10 2010-11-02 Lantheus Medical Imaging, Inc. Methods of making radiolabeled tracers and precursors thereof
US8926945B2 (en) 2005-10-07 2015-01-06 Guerbet Compounds comprising a biological target recognizing part, coupled to a signal part capable of complexing gallium
US8986650B2 (en) 2005-10-07 2015-03-24 Guerbet Complex folate-NOTA-Ga68
US8551448B2 (en) 2006-05-18 2013-10-08 The Regents Of The University Of California Rotanone analogs: method of preparation and use
WO2007137135A2 (fr) * 2006-05-18 2007-11-29 The Regents Of The University Of California Analogues de roténone, méthode de préparation et d'utilisation
WO2007137135A3 (fr) * 2006-05-18 2008-04-03 Univ California Analogues de roténone, méthode de préparation et d'utilisation
WO2008133730A2 (fr) * 2006-11-15 2008-11-06 The Regents Of The University Of California Analogues de roténone marqués
WO2008133730A3 (fr) * 2006-11-15 2009-02-05 Univ California Analogues de roténone marqués
US9408927B2 (en) 2008-02-29 2016-08-09 Lantheus Medical Imaging, Inc. Contrast agents for applications including perfusion imaging
US10245332B2 (en) 2008-02-29 2019-04-02 Lantheus Medical Imaging, Inc. Contrast agents for applications including perfusion imaging
US9687571B2 (en) 2009-04-15 2017-06-27 Lantheus Medical Imaging, Inc. Stabilization of radiopharmaceutical compositions using ascorbic acid
WO2010127054A3 (fr) * 2009-04-30 2011-01-06 Ge Healthcare Limited Imagerie d'une perfusion de tumeur et du métabolisme oxydatif à l'aide d'ace pet dynamique chez des patients atteints du cancer de la tête et du cou durant une radiothérapie
US9603951B2 (en) 2010-02-08 2017-03-28 Lantheus Medical Imaging, Inc. Methods and apparatus for synthesizing imaging agents, and intermediates thereof
US10022462B2 (en) 2010-02-08 2018-07-17 Lantheus Medical Imaging, Inc. Methods and apparatus for synthesizing imaging agents, and intermediates thereof
US8936777B2 (en) 2010-02-08 2015-01-20 Lantheus Medical Imaging, Inc. Methods and apparatus for synthesizing imaging agents, and intermediates thereof
US10842892B2 (en) 2010-02-08 2020-11-24 Lantheus Medical Imaging, Inc. Methods and apparatus for synthesizing imaging agents, and intermediates thereof
US9579408B2 (en) 2011-02-11 2017-02-28 Washington University PET/SPECT agents for applications in biomedical imaging
CN102336764B (zh) * 2011-06-21 2013-10-30 兰州大学 自旋标记鱼藤酮肟酯及其制备方法和用途
CN102336764A (zh) * 2011-06-21 2012-02-01 兰州大学 自旋标记鱼藤酮肟酯及其制备方法和用途
EP2731952A1 (fr) * 2011-07-13 2014-05-21 Nordion (Canada) Inc. Dérivés de roténone radio-marqués et leur utilisation en imagerie spect
EP2731952A4 (fr) * 2011-07-13 2015-01-07 Nordion Canada Inc Dérivés de roténone radio-marqués et leur utilisation en imagerie spect
JP2014520821A (ja) * 2011-07-13 2014-08-25 ノルディオン （カナダ） インク． 放射標識されたロテノン誘導体及びこれらのｓｐｅｃｔ画像化における使用
WO2013006958A1 (fr) 2011-07-13 2013-01-17 Nordion (Canada) Inc. Dérivés de roténone radio-marqués et leur utilisation en imagerie spect
US9919064B2 (en) 2012-08-10 2018-03-20 Lantheus Medical Imaging, Inc. Compositions, methods, and systems for the synthesis and use of imaging agents
US10500293B2 (en) 2012-08-10 2019-12-10 Lantheus Medical Imaging, Inc. Compositions, methods, and systems for the synthesis and use of imaging agents
US9713651B2 (en) 2012-08-10 2017-07-25 Lantheus Medical Imaging, Inc. Compositions, methods, and systems for the synthesis and use of imaging agents
US11744906B2 (en) 2012-08-10 2023-09-05 Lantheus Medical Imaging, Inc. Compositions, methods, and systems for the synthesis and use of imaging agents
US9850183B2 (en) 2014-06-27 2017-12-26 Reiley Pharmaceuticals, Inc. Conjugates derived from non-steroidal anti-inflammatory drugs and methods of use thereof in imaging
US10053478B2 (en) 2015-01-09 2018-08-21 Reiley Pharmaceuticals, Inc. COX-2-targeting, platinum-containing conjugates and their use in the treatment of tumors and cancers
US20210024555A1 (en) * 2018-03-22 2021-01-28 National Centre For Scientific Research "Demokritos" Tricarbonyl complexes of transition metals with benzo-heterocyclic derivatives of the cyclopentadienyl anion
US11845763B2 (en) * 2018-03-22 2023-12-19 National Centre For Scientific Research “Demokritos” Tricarbonyl complexes of transition metals with benzo-heterocyclic derivatives of the cyclopentadienyl anion
WO2022272268A1 (fr) * 2021-06-23 2022-12-29 Yale University Composés pour l'imagerie moléculaire du renouvellement de collagène et méthodes les utilisant

Also Published As

Publication number Publication date
AU2003224747A1 (en) 2003-10-27

Similar Documents

Publication Publication Date Title
US7875258B2 (en) Technetium-dipyridine complexes, and methods of use thereof
US8685369B2 (en) Technetium- and rhenium-bis (heteroaryl) complexes, and methods of use thereof
WO2003086476A1 (fr) Derives de rotenone a marquage au technetium, et procedes d'utilisation correspondants
EP2921482B1 (fr) Inhibiteurs marqués de l'antigène membranaire spécifique de la prostate (psma), évaluation biologique, et utilisation en tant qu'agents d'imagerie
US20060093552A1 (en) Technetium-and rhenium-bis(heteroaryl) complexes, and methods of use thereof
US7824661B2 (en) Technetium- and rhenium-bis(heteroaryl) complexes, and methods of use thereof
CA2400856C (fr) Agents d'imagerie pour le diagnostic de la maladie de parkinson
US8697032B2 (en) Prosthetic groups attached to stannyl polymer in the synthesis of radiopharmaceuticals
CN1972718B (zh) 锝-和铼-双（杂芳基）络合物，及其使用方法
US20100303725A1 (en) Technetium- and Rhenium-Bis(heteroaryl) Complexes, and Methods of Use Thereof
AU2012200218A1 (en) Technetim-and rhenium-bis(heteroaryl) complexes, and methods of use thereof

Legal Events

Date Code Title Description
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 CO CR CU CZ DE DK DM DZ EC 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 OM PH PL PT RO RU SC SD SE SG SK SL TJ TM TN TR TT TZ UA UG US UZ VC VN YU ZA ZM 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 ZM ZW AM AZ BY KG KZ MD RU TJ TM AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HU IE IT LU MC NL PT RO SE SI SK TR BF BJ CF CG CI CM GA GN GQ GW ML MR NE SN TD TG

121 Ep: the epo has been informed by wipo that ep was designated in this application
122 Ep: pct application non-entry in european phase
NENP Non-entry into the national phase

Ref country code: JP

WWW Wipo information: withdrawn in national office

Country of ref document: JP