Non-Isotopic Detection of Osteoblastic Activity In Vivo Using Modified Bisphosphonates

Abstract

The present invention is directed to a non-isotopic methods for the in vitro and in vivo detection of hydroxyapatite-positive cells and structures.

1 . A method for in vivo imaging comprising:
 (i) administering to an animal a contrast agent comprising a fluorescent moiety covalently linked to a bisphosphonate moiety;   (ii) obtaining a fluorescent image of the animal, or at least a portion thereof, at one or more wavelengths which detect the contrast agent;   (iii) constructing an image of the animal including the pattern of distribution of the contrast agent.   
   
   
       2 . The method of  claim 1 , wherein the fluorescent moiety has an excitation and emission spectra between about 650 nm and about 1000 nm. 
   
   
       3 . The method of  claim 1 , wherein the fluorescent moiety is a near-infrared fluorescent dye. 
   
   
       4 . The method of  claim 3 , wherein the near-infrared fluorescent dye is a cyanine dye. 
   
   
       5 . The method of  claim 1 , wherein the bisphosphonate moiety is independently selected from the group consisting of alendronate, clodronate, EB-1053, etidronate, ibandronate, incadronate, neridronate, olpadronate, phosphonate, pamidronate, risedronate, tiludronate, YH 529 and zoledronate. 
   
   
       6 . The method of  claim 1 , for evaluating the condition of one or more bones. 
   
   
       7 . The method of  claim 1 , for detecting bone diseases. 
   
   
       8 . The method of  claim 1 , for detecting metastases of cancer to bone. 
   
   
       9 . The method of  claim 1 , for evaluating anti-tumor therapies. 
   
   
       10 . The method of  claim 1 , for detecting blood vessel micro-calcifications. 
   
   
       11 . The method of  claim 1 , for evaluating one or more bones for a biomechanical property. 
   
   
       12 . The method of  claim 1 , as part of a protocol for diagnosing osteoporosis. 
   
   
       13 . The method of  claim 1 , for diagnostic detection of diseases accompanied with an abnormality of calcium hydroxyapatite. 
   
   
       14 . The method of  claim 1 , wherein the animal is a human. 
   
   
       15 . A method for in vivo imaging comprising:
 (i) administering to an animal a contrast agent comprising a near-infrared fluorescent dye covalently linked to a bisphosphonate moiety;   (ii) obtaining a fluorescent image of the animal, or at least a portion thereof, at one or more wavelengths which detect the contrast agent;   (iii) constructing an image of the animal including the pattern of distribution of the contrast agent.   
   
   
       16 . The method of  claim 15 , wherein the near-infrared fluorescent dye has an excitation and emission spectra between about 650 nm and about 1000 nm. 
   
   
       17 . The method of  claim 15 , wherein the near-infrared fluorescent dye is a cyanine dye. 
   
   
       18 . The method of  claim 15 , wherein the bisphosphonate moiety is independently selected from the group consisting of alendronate, clodronate, EB-1053, etidronate, ibandronate, incadronate, neridronate, olpadronate, phosphonate, pamidronate, risedronate, tiludronate, YH 529 and zoledronate. 
   
   
       19 . The method of  claim 15 , for evaluating the condition of one or more bones. 
   
   
       20 . The method of  claim 15 , for detecting bone diseases. 
   
   
       21 .- 27 . (canceled)