Compounds for targeting endothelial cells, compositions containing the same and methods for their use

Abstract

The present invention provides compounds for targeting endothelial cells, tumor cells or other cells that express the NP-1 receptor, compositions containing the same and methods for their use. Additionally, the present invention includes diagnostic, therapeutic and radiotherapeutic compositions useful for visualization, therapy or radiotherapy.

1 - 84 . (canceled) 
     
     
         85 . A composition for use in targeting endothelial cells, tumor cells or other cells which express NP-1, which comprises a compound of the formula (I)
   A-L-B  (I)
   in which   A is a monomer, a multimer or polymer of TKPPR (SEQ. ID. NO:2) or a monomer, multimer or polymer of a TKPPR (SEQ ID NO:2) analogue which contains TKPPR (SEQ ID NO:2) and which specifically binds to NP-1 or cells that express NP-1 with avidity that is equal to or greater than TKPPR (SEQ ID NO:2);   L is a linker; and   B is a substrate selected from a microparticle, a bead or a polymer which can be used to produce a microparticle or bead.   
     
     
         86 . A composition according to  claim 85 , wherein A is a tetramer of TKPPR or of a TKPPR analogue as defined in  claim 85 . 
     
     
         87 . A composition according to  claim 85 , wherein A is a monomer of TKPPR or a TKPPR analogue as defined in  claim 85 . 
     
     
         88 . A composition according to  claim 85 , wherein B is a microparticle optionally coated with one or more polymers to alter its surface properties. 
     
     
         89 . A composition according to  claim 85 , in which B is a bead. 
     
     
         90 . A composition according to  claim 85 , in which B further comprises a detectable label. 
     
     
         91 . A composition according to  claim 90 , in which the detectable label generates light, a radioactive signal or contains an enzyme or other signal generating system. 
     
     
         92 . A composition according to  claim 91 , in which the detectable label is a fluorescent dye. 
     
     
         93 . A composition according to  claim 85 , in which L is a bond or is derived from:
 an alkyl chain C 1 -C 6000 , linear or branched, saturated or unsaturated, optionally interrupted or substituted by one or more groups such as: O, S, NR, OR, SR, COR, COOH, COOR, CONHR, CSNHR, C═O, S═O, S(═O) 2 , P═O(O) 2 OR, P(O) 2 (OR) 2 , halogens, or phenyl groups, optionally substituted by one or more —NHR, —OR, —SR, —COR, —CONHR, —N—C═S, —N—C═O, halogens, in which   R is H or an alkyl group C 1 -C 4 , linear or branched, optionally substituted by one or more —OH;   such a chain can be interrupted or substituted by one or more cyclic groups C 3 -C 9 , saturated or unsaturated, optionally interrupted by one or more O, S or NR; by one or more groups such as: —NHR, —OR, —SR, —COR, —CONHR, or a phenyl group optionally substituted by one or more —NHR, —OR, —SR, —COR, —CONHR, —N—C═S, —N—C═O, halogens.   
     
     
         94 . A composition according to  claim 93 , in which the cyclic groups present in L are saturated or unsaturated, and correspond to the following general formula (III) 
       
         
           
           
               
               
           
         
         in which 
         n is 0 to 4; 
         m is 0 to 2; 
         X is NH, NR, O, S or SR. 
       
     
     
         95 . A composition according to  claim 91 , in which the linker L is an oligopeptide comprising 1 to 100 natural or synthetic amino acids. 
     
     
         96 . A composition according to  claim 93 , in which the amino acids are selected from the group consisting of glycine, glutamic acid, aspartic acid, γ-amino-butyric acid and trans-4-aminomethyl-cyclohexane carboxylic acid. 
     
     
         97 . A composition according to  claim 91 , in which L is derived from difunctional PEG (polyethyleneglycol) derivatives. 
     
     
         98 . A composition according to  claim 91 , in which L is selected from the group consisting of: glutaric acid, succinic acid, malonic acid, oxalic acid and PEG derivatized with two CH 2 CO groups. 
     
     
         99 . A composition according to  claim 89 , wherein B is a fluorescent bead. 
     
     
         100 . A composition according to  claim 97 , wherein A is tetramer of TKPPR or a TKPPR analogue as defined in  claim 85 . 
     
     
         101 . A contrast agent comprising a composition according to claim  84 , wherein B comprises a fluorescent label. 
     
     
         102 . The contrast agent of  claim 99  wherein A is a tetramer of TKPPR or a TKPPR analogue. 
     
     
         103 . A process for preparing a compound of claim  84  comprising:
 a) obtaining a monomer or multimer of TKPPR or a TKPPR analogue as defined in claim  1 ; 
 b) conjugating the monomer or multimer of TKPPR or TKPPR analogue with the linker to give a compound of formula (IIb); and
   A-L  (IIb)
 
 
 c) forming a covalent or non-covalent bond between a compound of formula (IIb) and the substrate B or forming a covalent bond between the substrate B and the linker to form a conjugate B-L, and 
 conjugating the conjugate B-L with the monomer or multimer of TKPPR or TKPPR analogue. 
 
     
     
         104 . A process according to  claim 101 , in which the compounds of formula (IIb) are prepared as illustrated in the following Scheme 
       
         
           
           
               
               
           
         
         in which 
         the steps a), b), and c) are all condensation reactions performed under basic conditions, and step d) is a condensation in basic conditions with the linker. 
       
     
     
         105 . A composition for use in targeting endothelial cells, tumor cells or other cells which express NP-1, which comprises a compound of the formula (I)
   A-L-B  (I)
   in which   A is a monomer, multimer or polymer of TKPPR (SEQ. ID. NO:2) or a monomer, multimer or polymer of a TKPPR (SEQ ID NO:2) analogue which contains TKPPR (SEQ ID NO:2) and which specifically binds to NP-1 or cells that express NP-1 with avidity that is equal to or greater than TKPPR (SEQ ID NO:2);   L is a linker; and   B is a fluorescent label.   
     
     
         106 . A composition according to  claim 105 , wherein A is a tetramer of TKPPR or a TKPPR analogue as defined in  claim 105 . 
     
     
         107 . A composition according to  claim 106 , wherein A is BRU 346. 
     
     
         108 . A composition according to  claim 105 , wherein B is a fluorescent dye. 
     
     
         109 . A composition according to  claim 107 , wherein B is Oregon Green. 
     
     
         110 . A method of imaging a NP-1-expressing angiogenic site in a human or animal comprising:
 a) administering to said human or animal an ultrasound contrast agent comprising a suspension of gas-filled microbubbles in which the microbubbles comprise a compound of the formula (I):
   A-L-B  (I)
 
   in which
 A is a multimer or polymer of TKPPR (SEQ ID NO:2) or a multimer or polymer of a TKPPR (SEQ ID NO:2) analogue which specifically binds to NP-1 or cells which express NP-1 with avidity that is equal to or greater than TKPPR (SEQ ID NO:2); 
 L is a linker; and 
 B is a phospholipid; and 
   b) detecting said contrast agent.   
     
     
         111 . A method of imaging endothelial cells, tumor cells or other cells that express NP-1 in a human or animal comprising:
 a) administering to said human or animal an ultrasound contrast agent comprising a suspension of gas-filled microbubbles in which the microbubbles comprise a compound of the formula (I):
   A-L-B  (II)
 
   in which
 A is a multimer or polymer of TKPPR (SEQ ID NO:2) or a multimer or polymer of a TKPPR (SEQ ID NO:2) analogue which specifically binds to NP-1 or cells which express NP-1 with avidity that is equal to or greater than TKPPR (SEQ ID NO:2); and 
 B is a phospholipid; and 
   b) detecting said contrast agent.   
     
     
         112 . The method of  claim 111 , wherein a tumor in a human or an animal is staged. 
     
     
         113 . The method of any one of  claims 110 - 112 , wherein A is a tetramer of TKPPR or an analogue of TKPPR. 
     
     
         114 . A method of treating an individual exhibiting effects of an NP-1 mediated or related disorder comprising administering to said human or animal an ultrasound contrast agent comprising a suspension of gas-filled microbubbles in which the microbubbles comprise a compound of the formula (I) and optionally a therapeutic moiety:
   A-L-B  (II)
   in which
 A is a multimer or polymer of TKPPR (SEQ ID NO:2) or a multimer or polymer of a TKPPR (SEQ ID NO:2) analogue which specifically binds to NP-1 or cells which express NP-1 with avidity that is equal to or greater than TKPPR (SEQ ID NO:2); and 
 B is a phospholipid.