MXPA99010058A - Radionuclide associated with nucleotide polyphosphate as tumor imaging agents - Google Patents

Abstract

The invention provides tumor imaging agents comprising a radionuclide in association with a nucleotide polyphosphate targeting molecule. Methods for using the tumor imaging agents and kits containing the tumor imaging agents or components suitable for production of the tumor imaging agents are also provided.

Description

RADIONÚCLIDO ASSOCIATED WITH POLYPHOSPHATE OF NUCLEOTIDES AS AGENTS OF REPRODUCTION OF TUMOR IMAGES The present invention relates to the field of nuclear medicine. More specifically, the invention relates to the reproduction of images for tumor diagnosis. BACKGROUND OF THE INVENTION The technology of clinical image reproduction plays a significant role in the diagnosis of damage and ase processes. Many parts of the human body can now be examined for diagnostic purposes. Many parts of the human body can now be examined for diagnostic purposes using a variety of imaging techniques. For a long time, radiography has been used to represent by body parts images through which generated X-rays are transmitted. Computed tomography (CT) provides X-ray images in cross section of a body plane. It can be targeted to specific tissues or organs in positron emission tomography (PET), single photon emission computed tomography (SSCT), and gamma scintigraphy. In PET, TCESF, and gamma scintigraphy, radiopharmaceutical agents capable of being sequestered (concentrated) to some degree in the tissue or target organ are internally administered to the patient and the images are generated by detecting the radioactive emissions of the concentrated radiopharmaceutical agent. Some of the radiopharmaceuticals currently used for image reproduction include nuclides such as 201TI, 99mTc, 233Xe, and the like; chelates of nuclides, radiolabelled metabolic agents such as 11D-deoxy-D-glucose, 8F-2-fluorodeoxy-D-glucose, fatty acid analogues of [1-11C] - and [123l] -β-methyl, 113N-ammonia , and the like; infarct-like agents such as 99mTc-tetracycline, 99mTc-pyrophosphate, 203-Hg-mercurial, 67Ga-citrate and the like, and radiolabeled ligands, proteins, peptides and monoclonal antibodies. Whole cells such as erythrocytes, platelets, leukocytes, and other cells can also be labeled with a radionuclide and function as radiopharmaceuticals. The amount and type of clinical information that can be derived from PET, TCESF, gamma scintigraphic images refers in part to the ability to concentrate radiopharmaceutical agent in the target tissue or organ. Although many radiopharmaceuticals are available for clinical use, the resolution of the generated image may be limited depending on several factors. The resolution of a particular image reproduction agent to represent images of dead or damaged tissue depends, in part, on the affinity of the radiopharmaceutical for the site of damage or ase, compared to its affinity for surrounding healthy tissue. Radiopharmaceuticals are used to diagnose and treat tumors. D.R. Elmaleh, and others (1984) Proc. Nati Acad. Sci. USA 81, 918-921 describes Ap4A labeled with 99mTc (99 Tc-Ap4A), used to represent tumor images implanted in rats. The method used to chelate 99mTc to Ap A in this study produced a mixture, in which 99mTc was bound to Ap A-dinucleotide and which may also contain 99 Tc unchelated. This study was based on the premise that some human tumor cells are permeable to exogenous ATP and ADP and that these cells are incorporated into intact nucleotides in intracellular combinations, in contrast to normal cells. Ap4A was shown to permeate in hepatoma cells but not in a number of cell lines of non-transformed mammals. In addition to accumulating in implanted tumors, in the 1984 study, 99 Tc-Ap4A also accumulated in the kidney, liver, bone, muscle and lung. SUMMARY OF THE INVENTION Radionuclide-labeled nucleotide polyphosphates accumulate with high specificity in tumors, and one embodiment of the invention generally characterizes tumor imaging agents that include a radionuclide associated with a nucleotide polyphosphate, the latter being a moiety that targets the target. The image reproduction agent has an improved ratio of radioactivity from target directed to non target target, as a result of the use of a co-eluent such as mannitol. In another embodiment, the invention provides a tumor imaging agent that includes a radionuclide associated with a targeting portion of polyphosphate nucleotides. The imaging agent is typically eluted with an eluent as described herein, and the agent formulation may include traces of that agent. Normally, the target portion is a residue of a precursor that targets the target; for example, a precursor targeting the target is reacted with a tagging entity that includes the radionuclide and a chelator for the radionuclide. The image reproduction agent is the reaction product that includes a residue of the target targeting precursor and the chelator, in association with the radionuclide. The association may involve one or more of: chelation, covalent binding or electrostatic binding, or it may involve other forces or combination of forces that the nucleoside holds in spatial proximity to a targeting molecule. The image reproducing agent may be the reaction product of the targeting precursor to the target with a portion containing radionuclides and said reaction may involve the formation of a chelate or a covalent reaction product, or a product in which both are involved. chelation as covalent bonds. Normally, the targeting precursor is a molecule of formula A) or formula B), or dimers or trimers thereof such as the molecules of formulas C) or D): A) Nu? - (p) mX B) Nu1- (p) "- X- (p) m-Nu2 C) Nu? - (p)" - X? - (p) m-Nu2 I NU3- (p) r-X2- (p) q -NU4, D) NUi-ÍpJn-Xí-ÍpJm- Uz X3 Nu3- (p) p-X2- (p) q-Nu4l wherein (1) each of Nu? -Nu is an independently selected nucleoside; (2) p is selected from the group consisting of a phosphate moiety, a phosphorothioate moiety, an alkyl phosphonate moiety, a phosphorodithioate moiety, a phosphoramidate moiety, an aminoalkyl phosphoramidate moiety, an aminoalkyl phosphotriester moiety, an aminoalkyl phosphorothioamidate moiety and a portion of thiophosphate; (3) each of X, X, X, and X3 is selected from the group consisting of an alkyl group, a halogenated alkyl group, a nitrogen-containing alkyl group, a sulfur-containing alkyl group, an alkylene group, an halogenated alkylene group, an alkylene group containing nitrogen and an alkylene group containing sulfur; (4) (n + m) is from 2 to 8; and (r + q) is from 2 to 8. Other agents (or residues thereof reacted with target spray) to promote chelation or binding may be present in the image reproduction agent.

In a preferred formula according to B), X is not optional and NUI and Nu2 are the same and are adenosine, guanine, histidine, thymidine, uracil or inosine. Preferably, at least one of Nu! - (more preferably each of Nu -? - Nu4) is adenosine. The preferred portions for x (when present) are the alkyl portions or chloroalkyl portions and p is preferably a phosphate moiety. When X is not present, the structure containing radionuclides can be chelated via an oxygen atom of the phosphate (s). Preferably, the nucleoside is adenosine, p is a phosphate, n = 2, and m = 2. The radionuclide (Z) that is finally associated or formed in complex with the targeting precursor can be 131l, 99m-Tc,, 4F, oeGa, "Cu, y, ln, although aamTc is preferred. If the radionuclide is associated via an optional chelating structure (R), particularly for 99 Tc, the chelating structure can be a structure of -N3S2. , a structure of -NS3, a structure of -N4, a ¡nitrile, a hydrazine, a structure containing nicotinic acid (HYNIC, for its acronym in English) a structure containing 2-methylthionicotinic acid, a group containing phosphorus, or a carboxylate group In a specific embodiment, Z is 99mTc, and is part of a 99mtc complex having the following formula: Ad- (p) 2-CH- (p) 3-Ad R-Z Ad - (p) 2-CH- (p) 2-Ad where Ad is adenosine, p is PO2H, and R is a complex-forming moiety. The above agents can be administered to reproduce images of tumors in a mammal. A method for reproducing specific images detects tumors by administering the imaging agent to the mammal and detecting the spatial distribution of the agent. The differential accumulation of the agent indicates a tumor. The invention also characterizes equipment for reproducing tumor images comprising the image reproduction agent. The kit can include a chelating agent and / or an auxiliary molecule selected from the group consisting of mannitol, gluconate, glucoheptonate, and tartrate; and a tin-containing reducing agent, such as SnCl2 or tin tartrate. BRIEF DESCRIPTION OF THE DRAWINGS The foregoing and other objects of this invention, the different aspects thereof, as well as the invention itself, can be more easily understood from the following description, when read together with the accompanying drawings in which : Figure 1 shows a CLAR chromatogram of a reaction mixture obtained when 50 mg of mannitol was used as an auxiliary molecule. The peaks of the elution profile are extrapolated, with areas under the peaks of the chromatogram tabulated below the elution profile.

Figure 2 shows a CLAR chromatogram of an alternative reaction mixture, in which 10 mg of mannitol was used as an auxiliary molecule. The peaks of the elution profile were extrapolated, with the areas under the chromatogram peaks tabulated below the elution profile. Figure 3 is a set of radio-images of tumors in diseased mice. Detailed Description of the Preferred Modalities The patent and scientific literature referred to herein establishes the knowledge that it is available to those skilled in the art. The patents of E.U.A. and the permitted applications cited herein are incorporated herein by reference.

The present invention novel tumor imaging agents in which polyphosphate analogs of nucleotides are associated with a radionuclide. The imaging agents of the invention accumulate specifically in tumors. The image reproduction agents of the invention contain a targeting molecule which accumulates specifically in tumors. In general, preferred targeting molecules have the formula (or dimers or other multimers thereof) Nu- (p) nX- (p) m-Nu where Nu is a radionucleoside selected from the group consisting of adenosine, guanine , histidine, thymidine, uracil, and inosine; p is selected from the group consisting of a phosphate moiety, a phosphorothioate moiety, an alkyl phosphonate moiety, a phosphoryldioate moiety, a phosphoramidate moiety, an aminoalkyl phosphoramidate moiety, an aminoalkyl phopotriester moiety, and an aminoalkylphosphorothioamidate moiety; X is selected from the group consisting of an oxygen, an alkyl group, a halogenated alkyl group, a nitrogen-containing alkyl group, a sulfur-containing alkyl group, an alkylene group, a halogenated alkylene group, an alkylene group containing nitrogen, an alkylene group containing sulfur, and (n + m) is from 2 to 8. Formulations of more than one molecule that targets a target labeled with a nuclide. Any nucleoside can be used as the "Nu" component of the molecule that targets the target. In addition to the ribonucleosides listed above, the "Nu" component of the targeting molecule can be deoxynucleoside, an unsubstituted ribonucleoside, or a substituted deoxyribonucleoside, a substituted ribonucleoside, or a substituted deoxyribonucleoside. They can be presented in accordance with the invention in the base portion of nucleic acids or on the sugar portion of the nucleoside. Sugars other than ribose or deoxyribose may be present in the "Nu" component of the molecule that targets the target. Preferably, the nucleoside of the molecule targeting the target is adenosine.

Any molecule can be used as the "p" portion to bind the nucleoside components of the molecule that targets the target. The targeting molecule may contain synthetic internucleoside linking portions other than the "p" portions listed above, wherein the "p" portion is a substituted phosphate. For example, the "p" portion of substituted phosphate may contain one of more alkyl groups, carbamate groups, acetamidate groups, and the like. The "p" portion may additionally be a phosphorus-free group such as a carboxymethyl ester or a carbonate, while the conformation of the molecule targeting the resulting target approximates that of a regulatory dinucleoside such as Ap4A. Any portion can be used as the "X" component of the molecule targeting the target while the conformation of the molecule targeting the resulting target approaches that of a regulatory dinucleoside such as Ap4A or Ap A dimer. to synthesize the molecule that targets the target is usually displayed in GM Blackburn, et al. (1966) in Biofhosfhates and Their Analogues - Synthesis, Structure, Metal and activity, eds. Bruzik, K.S. & Stec, W.J. (Elsevier, Amsterdam) p. 451-464; G.M. Blackburn, et al., (1987) Nucleic Acids Res. 17, 6991-7004; A. Guranowski et al. 81987) Biochemistry 26, 3425-3429; and G.M. Blackburn, and others 81992) in Dinucleoside Polyphosphates, ed. McLennan, A.G. (CRC, Boca Ratón, Fl) Chapter According to the invention, the molecule that targets the target is associated with the nuclide (spatial proximity). The spatial proximity between the targeting molecule and the nuclide can be made in any way that preserves the specificity of the target target molecule for its target tissue. For example, the spatial proximity between the nuclide and the molecule that targets the target can be effected by a non-covalent chemical bond. So that a chemical bond can be made through a chelating substance and / or an auxiliary molecule such as mannitol, gluconate, glucoheptonate, tartrate and the like. Alternatively, the spatial proximity between the nuclide and the target molecule can be effected by incorporating the nuclide and the target molecule into a micelle or liposome, in such a way that the affinity of the target molecule is maintained. white for your white tissue. The spatial proximity between the nuclide and the targeting molecule can also be effected by linking the nuclide and the targeting molecule to a matrix such as a microsphere or liposomes. Those skilled in the art will recognize that there are a number of ways to synthesize imaging agents. In one synthesis, Nu-p2-CHCI-p2Nu is reacted with X-R, wherein X is defined as X? -3, before, and R is a radionuclide or a radionuclide chelating agent such as a 99mTc chelating agent. Normal 99mTc chelating agents include allyl or aryl amines or alkyl or aryl thiols. Other chelating groups of 99mTc include N2S2, S4. A precursor containing the chelating agent -R, which in turn contains the chelating agent -R, which in turn is reacted with 99mTc of a normal 99mTc reduction equipment. For example, when -R is an alkyl lime, the chelating reaction of -NS3 forms: Nu-p2-CH "p2 ~ Wu INCH2) n S 0 = w" TC »0 (CHa) -S S- (CH,) N If the composition includes a chelating structure, particularly for 99mTc, the chelating structure can be a structure of N2S2, a structure of -NS3, a structure of N4, an isonitrile, a hydrazine, a group of HYNIC (hydrazinonicotinic acid), a group which contains phosphorus, or a carboxylate group. Those skilled in the art will understand that many other chelation reactions can be used. The image reproduction agents described above may contain any radionuclide according to the invention. Preferably, the image reproduction agents of the invention contain radionuclides suitable for use in PET or TCESF that reproduce images. More preferably, the radionuclide (Z) used in the imaging agent is a radionuclide selected from the group consisting of 1231, 99mTc, 18F, 68Ga, 62Cu, 111ln, and the like. Said radionuclides can be incorporated in the image reproduction agent by covalent binding directly to an atom of the target molecule, or the radionuclide can be non-covalently or covalently associated with the molecule targeting the target through a chelating structure or through an auxiliary molecule such as mannitol, gluconate, glucoheptonate, tartrate and the like. When a chelating structure is used to provide spatial proximity between the radionuclide and the targeting molecule, the chelating structure can be directly associated with the targeting molecule or can be associated with the molecule targeting the target through an auxiliary molecule such as mannitol, gluconate, glucoheptonate, tartrate and the like. Any suitable chelating structure can be used to provide spatial proximity between the radionuclide and the molecule targeting the agent through non-covalent association. Many chelating structures are known in the art. Preferably, the chelating structure is a structure of N2S2, a structure of NS3, a structure of N, a structure containing isonitrile, a structure containing hydrazine, a structure containing the group of HYNIC (hydrazinonicotinic acid), a structure which it contains a group of 2-methylthionicotinic acid, a structure containing a carboxylate group, and the like. In some cases, chelation can be achieved without including a separate chelation structure, because the radionuclide directly chelates the atoms in the targeting portion, for example, to oxygen atoms in the phosphate groups or carboxylate groups . The chelation structure, the auxiliary molecule, or the radionuclide can be placed in spatial proximity for any position of the molecule that targets the target that does not interfere with the interaction of the target molecule with its receptor in tumors. The chelation structure, auxiliary molecule, or radionuclide can be covalently or non-covalently associated with any portion of the molecule that targets the target except the receptor binding portion. For example, the chelation structure, the auxiliary molecule, or radionuclide can associate with the phosphate portion of I to target molecule, with the -X- portion of the molecule targeting the target. The radionuclides can be placed in spatial proximity to the molecule targeting the target using known methods that effect or optimize the chelation, association or binding of the specific radionuclide to the ligands. For example, when 1231 is the radionuclide, the imaging agent can be labeled according to known radioiodination procedures such as direct radioiodination with chloramine T, radioiodination exchange for a halogen or an organometallic group, and the like. When the radionuclide is 99rnTc, the imaging agent can be labeled using any suitable method to bind 99mTc to a ligand molecule. Preferably, when the radionuclide is 99mTc, an auxiliary molecule such as mannitol, gluconate, glucoheptonate, or tartrate is included in the labeling, cone reaction mixture without a chelating structure. More preferably, 99mTc is placed in spatial proximity to the molecule targeting the target by reducing 99mTCO 4 with tin in the presence of mannitol and the molecule targeting the target. When Ap A or an analogue of Ap4A is the molecule that targets the target, it was preferably done by reducing approximately fifty to 100 mCi of 99 TcO4 with 0.05-1 mg SnCl2 in the presence of approximately 1 to 20 mg of mannitol, for each mg of or similar to Ap4A. More preferably, for each mg of Ap A or analog of Ap4A, about 0.05 mg SnCl2 and about 10 mg of mannitol are used to reduce 99mTcO. Other reducing agents, e.g., tin tartrate, may also be used to form the image reproduction agent of the invention. An illustrative procedure for placing 99mTc in spatial proximity to Ap A and the Ap A analogue in which the "X" portion is -CHCI- is shown in Example 1. After the labeling reaction is completed, the mixture of reaction can optionally be purified using one or more steps of high performance liquid chromatography (HPLC). Any suitable CLAR system can be used if a purification step, and the performance or image reproduction agent of the invention is carried out. For example, the pH may vary, e.g., arise to decrease the elution time of the peak corresponding to the imaging agent of the invention. The invention is modalized in an equipment for reproducing images comprising one or more of the imaging agents described above, in combination with a pharmaceutically acceptable carrier such as human serum albumin. Human serum albumin for use in the kit of the invention can be made in any form, for example, through the purification of human serum protein or through the recombinant expression of a vector that contains a gene encoding albumin of human serum. Other substances can also be used as carriers according to this embodiment of the invention, for example, detergents, diluted alcohols, carbohydrates, auxiliary molecules and the like. The equipment of the invention, of course, may also contain other articles that may facilitate its use, such as syringes, instructions, reaction flasks and the like. In one embodiment, a kit according to the invention contains from about 1 to about 30 mCi of the radionuclide-labeled tumor imaging agent described above, in combination with a pharmaceutically acceptable carrier. The kits of the invention can include Ap A analogs labeled with radionuclides. The agent for reproducing tumors and vehicle images can be provided in solution or in lyophilized form. When the tumor imaging agent and vehicle of the kit are in lyophilized form, the kit may optionally contain a sterile and physiologically acceptable reconstitution medium such as water, saline, saline with regulated pH, and the like. In another embodiment, the kit of the invention may contain the molecule targeting the unlabeled target which has been combined covalently or non-covalently with a chelating agent.; an auxiliary molecule such as mannitol, gluconate, glucoheptonate, tartrate and the like; and a reducing agent such as SnCl2 or tin tartrate. In this mode, the equipment may include unmarked Ap4A or an unmarked Ap4A analog. The targeting molecule / unlabeled chelating agent, the auxiliary molecule and the reducing agent can be provided in solution or in lyophilized form and these components of the equipment of the invention optionally can contain stabilizers such as NaCl, silicate, phosphate buffer solutions, ascorbic acid, gentisic acid and the like. Further stabilization of the equipment components can be provided in this mode, for example, by providing the reducing agent in an oxidation-resistant form. The determination and optimization of said stabilizers and stabilization methods are within the level of experience of the art. When the targeting molecule / chelating agent not lauded in this embodiment has a lyophilized form, the kit may optionally contain a sterile and physiologically acceptable reconstitution medium, such as water, saline, saline with regulated pH, and the like. The amounts of the unlabeled site / chelating agent molecule, helper molecule, and reducing agent in this embodiment are optimized according to the methods for forming the tumor imaging agent set as set forth above. Radionuclides, such as 99mTc from a commercially available Mo / Te generator or 123, I commercially available, can be combined with the targeting molecule / unlabeled chelating agent and the reducing agent for a time and at a temperature sufficient to chelate the radionuclide to the targeting molecule / chelating agent and the imaging agent thus formed was injected into the patient. The tumor imaging agents of the invention can be used according to the methods of the invention by those skilled in the art, e.g., by nuclear medicine specialists, to reproduce a tissue image in a mammal. Any mammalian tumor can be reproduced in the image imaging agents of the invention are suitable for reproducing the images of breast tumors, melanomas of prostate tumors, colon tumors, etc. The methods of the invention can employ Ap4A labeled with radionuclides or an Ap A analog labeled with radionuclides. The images are generated by virtue of the differences in the spatial distribution of the pre-imaging images that accumulate in the different tissues or organs of the mammal. The spatial distribution of the imaging agent accumulated in a mammal in an organ, or in a tissue can be measured using any suitable means, for example, a gamma camera, a PET apparatus, a TCEUF apparatus, and the like. Some tumors may be evident when a less intense point appears within the image, for example, within an Ap4A labeled with 99mTc that indicates the presence of tissue in which a lower concentration of image reproduction agent accumulates relative to the concentration of imaging agent which accumulates in the tumor. Alternatively, a tumor may be detected as a more intense spot within the image, indicating a region of increased concentration of the image-reproducing agent in the tumor relative to the concentration of the agent that accumulates in the surrounding tissue. The degree of accumulation of the image reproduction agent can be quantified using known methods for quantifying radioactive emissions. A particularly useful image reproduction approach employs more than one margin reproduction agent to carry out simultaneous studies. The margin-forming agents of the invention are used in the following manner. An effective amount of an imaging agent comprising at least one targeting molecule and a nuclide (1 to 50 mCi) can be combined with a pharmaceutically acceptable carrier for use in imaging studies. According to the invention, "an effective amount" of the image reproduction agent of the invention is defined as an amount sufficient to give an acceptable image using equipment that is available for clinical use. An effective amount of the image reproducing agent of the invention can be administered in more than one injection. The effective amounts of the imaging agent of the invention will vary according to factors such as the degree of susceptibility of the individual, the age, sex and weight of the individual, idiosyncratic responses of the individual, dosimetry. The effective amounts of the imaging agent of the invention will also vary according to the instrument and factors related to the film. The optimization of these factors is within the level of experience in the field. As used herein, "pharmaceutically acceptable carrier includes any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic agents, absorption retardation agents and the like." The formulation used in the present invention may also contain stabilizers, preservatives, buffer solutions, antioxidants, or other additives known to those skilled in the art The use of such media and agents for pharmaceutically active substances is well known in the art.The supplementary active compounds can also be incorporated into the agent The imaging agent of the invention can be further administered to an individual in a suitable diluent or adjuvant, co-administered with enzyme inhibitors or in an appropriate vehicle such as human serum albumin or liposomes. pharmaceutically acceptable diluents they include sterile saline and other aqueous buffer solutions. The adjuvants contemplated herein include resorcinols, nonionic surfactants, such as polyoxyethylene oleyl ether and n-hexadecyl polyethylene ether. Enzyme inhibitors include pancreatic trypsin inhibitor, diethyl pyrocarbonate, and trasilol. Liposome inhibitors include water-in-oil emulsions in CGF water as well as conventional liposomes (Strejan et al., (1984) J. Neuroimmunol., 7, 27). Preferably, the imaging agent of the invention was administered intravenously and the imaging agent will be formulated as a sterile, pyrogen-free, parenterally acceptable aqueous solution. The preparation of said parenterally acceptable solutions, taking into account pH, isotonicity, stability and the like, are within the skill in the art. A preferred formulation for intravenous injection should contain, in addition to the imaging agent, an isotonic vehicle such as Sodium Chloride Injection, Ringer's Injection, Dextrose Injection, Injection of Dextrose and Sodium Chloride, Lactated Ringer's Injection, or another vehicle as is known in the art. The amount of the image reproduction agent used for diagnostic purposes and the duration of the image reproduction study will depend on the nature and severity of the condition being treated, the nature of the therapeutic treatments to which the patient has been subjected. patient, and the idiosyncratic responses of the patient. Finally, the doctor will decide the amount of imaging agent to administer to each individual patient and the duration of the image reproduction study. The following examples are included for illustrative purposes only and are not intended to limit the scope of the invention. EXAMPLE 1 SYNTHESIS OF 99mTc-Ap4A-binding (mannitol or gluconate) and 99mTc- Ap2CHCIp2A-binding (mannitol or gluconate) 50 to 100 mCi of 99mTc were obtained from a 99Mo / 99 Tc generator (DuPont Merk, Billerica, MA) in 1.5 ml of sterile water and mixed with a solution of one mg of Ap4A (Sígma, St. Louis, MO), 50 mg mannitol and 100 μg SnCl2 in 200 μg of sterile water. The solution was stirred and allowed to react for ten minutes at room temperature. The reaction mixture was then purified with HPLC by adsorption on a reversed phase column of C8-ODS 25 cm long and 5 mm in diameter (Waters, Milford, MA) and elution with CH3CN: pH 2.4 regulatory solution. Figure 1 shows an elution profile using this procedure with an unlabeled material shown by the black line (measured by A254), and the marked material shown in the clearest solid line (measured by t-emissions). The dotted line in Figure 1 is the elution profile of radioactivity. The radioactive elution peak at 15,826 minutes is 99mTc-Ap4A, which was injected into diseased mice as exhibited in Example 2, to reproduce the tissue images. The radioactive peak eluting at 1-3 minutes in this chromatogram was 99mTc-mannitol, which was used as a control in the experiments described in Example 2. A similar procedure was used to label the Ap4A analog of the formula: A- (p) 2-CHCl- (p) 2-A With 99mTc. Figure 2 is a reverse phase HPLC elution profile of a 99mTc-Ap A preparation in which 10 mg of mannitol and 40 μg of SNCI2 was used to carry out the reduction of one mg of Ap A. The reaction mixture was also stirred and allowed to react for ten minutes at room temperature. The purification of CLAR from the reaction mixture was carried out as before, except that the ratio of CH3CN: pH buffer was 15:85 and the pH buffer solution of elució contained 3.0 ml of concentrated H3PO4 and 3.36 my t-butylammonium hydroxide titrated with NaOH at pH 3.4. Figure 2 shows the elution profile of the reaction mixture, having a peak elution at 13,430 minutes corresponding to 99mTc-Ap4A.

When one mg of mannitol was used as an auxiliary molecule in the labeling reaction, very low yields of 99mTc-Ap4A were obtained. EJ EM PLO 2 Ap A was labeled with Tc-99m by reaction with Tc-99m gluconate and the product was purified by a reverse phase HPLC column C-8-9ds (0.5 x 25 cm). The product was eluted with CH3CN / pH buffer (20:80 by volume): The buffer solution was prepared by mixing H3PO4 (3.1 ml) with t-butylammonium hydroxide (3.9 ml) and the pH was adjusted to 2.4 by adding additional t-butylammonium oxide. The CLAR system that used Tc-99m-AP4A-glyco eluted with a retention time of 16 minutes. The radiochemical yields were 10-30% and the radiochemical purity was > 95% EX EMPLO 3 REPRODUCTION OF IMAGE IS OF TUMORS A tumor model was prepared by inoculating BT20 of human breast tumor cells in the shoulder region of diseased mice. When tumors reached 0.5 cm, groups of mice (n = 14) were injected with 0.5 m-Ci of TC-99m-AP4A-glyco or Tc-99, m-mannitol (control), prepared as described above. At 0, 5, 3 and 5 hours, gamma camera images were acquired and contralateral to tumor ratios were calculated. In 5 and 8 hours, the groups of 9 and 5 animals were sacrificed and biodistribution was measured. In all animals, the tumors were clearly visualized in 0.5 hours at T / S (ratio of tumor to blood) increased in the last images. In all three image representations, there was a very low level of trace accumulation in normal tissues. Biodistribution studies showed high concentrations (% ID / grams) of TC-99m-AP4-A-mannitol in tumors: 7.3 + 3.2 and 11.0 + 2.8 in 5 and 8 hours, respectively. In contrast, the concentration of Tc-99m-mannitol was 0.9%. Fig. 3 is an image produced generally using the prior art.

Claims (21)

1. CLAIMS 1. A tumor imaging agent comprising a radionuclide associated with a targeting portion, said target targeting portion comprising a polyphosphate residue of nucleotides.
2. 2. A tumor imaging agent comprising a radionuclide associated with a targeting portion, the target targeting portion comprising a polyphosphate precursor residue of nucleotides including formula A), B), C), or D): A) NUl- (p) mX B) Nu1- (p) "- X- (p) m-Nu2 C) Nu1- (p)" - X1- (p) m-Nu2 Nu3- (p) r-X2- (p) q-Nu4, D) Nu1- (p) "- X1- (p) m-Nu2 X
3. 3 Nu3- (p) p-X2- (p) q-Nu4, wherein (1) each of Nu -? - Nu is an independently selected nucleoside; (2) p is selected from the group consisting of a phosphate moiety, a phosphorothioate moiety, an alkyl phosphonate moiety, a phosphorodithioate moiety, a phosphoramidate moiety, an aminoalkyl phosphoramidate moiety, an aminoalkyl phosphotriester moiety, an aminoalkyl phosphorothioamidate moiety and a portion of thiophosphate; (3) each of X, Xi, X2, and X3 is selected from the group consisting of an alkyl group, a halogenated alkyl group, an alkyl group containing nitrogen, an alkyl group containing sulfur, an alkylene group, a group halogenated alkylene, an alkylene group containing nitrogen and an alkylene group containing sulfur; (4) (n + m) is from 2 to 8; and (r + q) is from 2 to 8. The agent of claim 2, wherein each of Nu2-Nu4 is independently selected from the group of nucleosides consisting of adenosine, guanine, histidine, thymidine, uracil and i n or s a n.
4. 4. The agent of claim 2, wherein at least one of Nu! -N is adenosine.
5. 5. The agent of claim 4, wherein each of Nu -? - Nu4 is adenosine.
6. 6. The agent of claim 1 or claim 2, wherein the radionuclide is selected from the group consisting of 123l, 99mt T "e, 18, F-, 68 ^ Ga", 62 C, u ,, 111 I l "n.
7. The agent of claim 2, wherein the image reproducing agent comprises the combination product of the targeting precursor with a chelating structure that chelates said radionuclide.
8. 8. The agent of claim 7, wherein the chelating structure is selected from the group consisting of a structure of H2S2, a structure of -NS3, a structure of -N, an isonitrile, a hydrazine, a structure containing a group of HYNIC , 2-metiothiolnicotinic acid group and a structure containing a carboxylate group.
9. 9. The agent of any of claims 7-8, wherein the radionuclide is 99mTc.
10. 10. The agent of any of claims 2-5, wherein x is an alkyl portion or a chloroalkyl portion and p is a phosphate moiety.
11. 11. A tumor imaging agent comprising a complex of 99 Tc: Ad- (p) 2-CH- (p) 3-Ad X Ad- (p) 2-CH- (p) 2-Ad where Ad is adenosine, and p is PO2H, and X is a chelating moiety containing 99mTc.
12. 12. A method for reproducing tumor images in a mammal comprising the steps of administering to the mammal the imaging agent of claim 1, claim 2, or claim 11.
13. 13. The method of claim 12, wherein the method detects a tumor in a mammal, the method comprising the steps of administering to the mammal the imaging agent, detecting the spatial distribution of the accumulated agent in the mammal.
14. A method for reproducing images of a tumor in a mammal comprising the steps of administering to the mammal an image reproduction agent according to claim 1, claim 2, or claim 11.
15. 15. The method of claim 14 , wherein the imaging agent is the imaging agent of claim 10.
16. 16. The method of claim 14, further comprising reacting the targeting precursor to the blank with a portion containing radionuclides.
17. 17. A device for reproducing tumor images comprising a supply of a targeting precursor to the target, the targeting precursor to the target having the formula A), B), C), or D): A) NUl- (p) m-X B) Nu1- (p) p-X- (p) m-Nu2 C) Nu1- (p) n -X1- (p) m-Nu2 Nu3- (p) r-X2- (p) q-Nu4, D) Nu? - (p) "- X? - (p) m-Nu2 X3 Nu3- (p) p-X2- (p) q- Nu4, wherein (1) each of Nu? -Nu4 is an independently selected nucleoside; (2) p is selected from the group consisting of a phosphate moiety, a phosphorothioate moiety, an alkyl phosphonate moiety, a phosphorodithioate moiety, a phosphoramidate moiety, an aminoalkyl phosphoramidate moiety, an aminoalkyl phosphotriester moiety, an aminoalkyl phosphorothioamidate moiety and a portion of thiophosphate; (3) each of X, X1f X2, and X3 is selected from the group consisting of an alkyl group, a halogenated alkyl group, a nitrogen-containing alkyl group, an alkyl group containing sulfur, an alkylene group, an alkylene group halogenated, an alkylene group containing nitrogen and an alkylene group containing sulfur; (4) (n + m) is from 2 to 8; and (r + q) is from 2 to 8.
18. The equipment of claim 17, further comprising a chelating agent, the chelating agent comprising an auxiliary molecule selected from the group consisting of mannitol, gluconate, glucoheptonate and tartrate; and b) a tin-containing reducing agent.
19. 19. The kit of claim 17, wherein at least one of NUÍ-N4 is adenosine.
20. 20. The kit of claim 19, wherein at least one of Nu? -N4 is adenosine. The equipment of claim 19, wherein said equipment further comprises at least one chelating agent. 24. The kit of claim 23, wherein the chelating agent is selected from the group consisting of a -N2S2 structure, a -NS3 structure, a -N4 structure, an isonitrile, a hydrazine, a structure containing a group of HYNIC, 2-metiothiolnicotinic acid group and a structure containing a carboxylate group. 25. The kit of any of claims 23 or 24, wherein the radionuclide is 99mTc. 26. The kit of any of claims 2-5, wherein x is an alkyl portion or a chloroalkyl portion and p is a phosphate moiety. 27. The kit of claim 17, wherein the nucleoside is adenosine, p is a phosphate, n = 2 and m = 2.