Nanoparticle peg modification with h-phosphonates

Abstract

The present invention provides phosphonate conjugates and methods of preparing the phosphonate conjugates so as to allow, for example, improved methods and compounds for modifying the surface of a nanoparticle to increase in vivo circulation times and targeted delivery performance.

1 - 20 . (canceled) 
     
     
         21 . A method of preparing a phosphonate conjugate, the method comprising:
 combining a primary amine compound having the formula: H 2 N(L 1 )-(R 1 ),   
       a carbonyl compound having the formula: O═C[(L 2 )-(R 2 )] n , and 
       a H-phosphonate compound having the formula: 
       
         
           
           
               
               
           
         
       
       under conditions sufficient to form the phosphonate conjugate having the formula: 
       
         
           
           
               
               
           
         
         wherein 
         each L 1 , L 2 , L 3  and L 4  is independently selected from the group consisting of a bond and a linking group; 
         R 1  is selected from the group consisting of a nanoparticle, an attachment component, a targeting agent, a diagnostic agent, a stealth agent, and a tetrapodal presentation component; 
         each R 2  is independently selected from the group consisting of a stealth agent, C 1 -C 10  alkyl, a carboxylic acid or ester, a phosphonic acid or ester, a sulfonic acid or ester, and a hydroxy; 
         each R 3  and R 4  is independently selected from the group consisting of H, an attachment component, a targeting agent, a diagnostic agent, and a stealth agent, wherein at least one of R 3  or R 4  is other than H; and 
         n is an integer from 0 to 2, wherein when n is 0 or 1 the carbonyl compound is an aldehyde. 
       
     
     
         22 . The method of  claim 21 , wherein the primary amine compound, the carbonyl compound, and the H-phosphonate compound are combined at a molar ratio of 1:2:2, respectively. 
     
     
         23 . The method of  claim 21 , wherein the primary amine compound and the carbonyl compound are combined before being combined with the H-phosphonate compound. 
     
     
         24 . The method of  claim 21 , wherein the H-phosphonate compound and the carbonyl compound are combined before being combined with the primary amine compound. 
     
     
         25 . The method of  claim 21 , wherein two of the H-phosphonate compounds are adjacently associated with a bilayer of a liposome before being combined with the primary amine compound and the carbonyl compound, wherein each of R 3  and R 4  is independently selected from the group consisting of a lipid and cholesterol. 
     
     
         26 . A compound of the formula: 
       
         
           
           
               
               
           
         
         wherein 
         each L 1 , L 2 , L 3  and L 4  is independently selected from the group consisting of a bond and a linking group; 
         R 1  is selected from the group consisting of a nanoparticle, an attachment component, a targeting agent, a diagnostic agent and a stealth agent; 
         R 2  is independently selected from the group consisting of a stealth agent, C 1 -C 10  alkyl, a carboxylic acid or ester, a phosphonic acid or ester, a sulfonic acid or ester, and a hydroxy; 
         each R 3  and R 4  is independently selected from the group consisting of H, a nanoparticle, an attachment component, a targeting agent, a diagnostic agent, and stealth agent, wherein at least one of R 3  or R 4  is other than H; 
         R 9  is a C 1 -C 10  alkyl group; and 
         n is an integer from 0 to 2. 
       
     
     
         27 . The compound of  claim 26 , wherein each of R 3  and R 4  is independently selected from the group consisting of a lipid and cholesterol; and each of L 3  and L 4  is a bond. 
     
     
         28 . The compound of  claim 27 , wherein the lipid is saturated or unsaturated C 10 -C 22  alkyl. 
     
     
         29 . The compound of  claim 26 , wherein R 1  is a stealth agent. 
     
     
         30 . The compound of  claim 29 , wherein the stealth agent is selected from the group consisting of PEG 100 , PEG 500 , PEG 1000 , PEG 2000 , PEG 5000  and PEG 10000 . 
     
     
         31 . The compound of  claim 26 , wherein R 9  is C 1 -C 6  alkyl and L 1  is a bond. 
     
     
         32 . A method of preparing a phosphonate conjugate, the method comprising:
 combining a secondary amine compound having the formula: HN[(L 1 )-(R 1 )](R 9 ), a carbonyl compound having the formula: O═C[(L 2 )-(R 2 )] n , and a H-phosphonate compound having the formula:   
       
         
           
           
               
               
           
         
         under conditions sufficient to form the phosphonate conjugate having the formula: 
       
       
         
           
           
               
               
           
         
         
           wherein 
           each L 1 , L 2 , L 3  and L 4  is independently selected from the group consisting of a bond and a linking group; 
           R 1  is selected from the group consisting of a nanoparticle, an attachment component, a targeting agent, a diagnostic agent and a stealth agent; 
           R 2  is independently selected from the group consisting of a stealth agent, C 1 -C 10  alkyl, a carboxylic acid or ester, a phosphonic acid or ester, a sulfonic acid or ester, and a hydroxy; 
           each R 3  and R 4  is independently selected from the group consisting of H, an attachment component, a targeting agent, a diagnostic agent, and a stealth agent, wherein at least one of R 3  or R 4  is other than H; 
           R 9  is selected from the group consisting of a C 1 -C 10  alkyl group; and 
           n is an integer from 0 to 2, wherein when n is 0 the carbonyl compound is a formaldehyde. 
         
       
     
     
         33 . (canceled) 
     
     
         34 . The targeted delivery composition of  claim 41 , wherein the nanoparticle is a liposome and each of R 3  and R 4  is associated with a bilayer of the liposome and independently selected from the group consisting of a lipid and cholesterol. 
     
     
         35 . (canceled) 
     
     
         36 . The targeted delivery composition of  claim 42 , wherein the nanoparticle is a liposome, and R 1  is associated with a bilayer of the liposome and independently selected from the group consisting of a lipid and cholesterol. 
     
     
         37 . A method for treating or diagnosing a cancerous condition in a subject, comprising administering to the subject a targeted delivery composition of  claim 41 , wherein the composition comprises a therapeutic or a diagnostic agent that is sufficient to treat or diagnose the condition, and wherein at least one of R 1 , R 3  and R 4  is a targeting agent. 
     
     
         38 . The method of  claim 37 , wherein the nanoparticle is a liposome and the therapeutic agent is encapsulated in, embedded in, or tethered to the liposome. 
     
     
         39 . The method of  claim 38 , wherein the therapeutic agent is an anticancer agent selected from the group consisting of doxorubicin, cisplatin, oxaliplatin, carboplatin, 5-fluorouracil, gemcitibine and a taxane. 
     
     
         40 . A method of determining the suitability of a subject for a targeted therapeutic treatment, comprising administering to the subject a targeted delivery composition of  claim 41 , wherein at least one of R 1 , R 3  and R 4  is a targeting agent and the nanoparticle comprises a diagnostic agent, and imaging the subject to detect the diagnostic agent. 
     
     
         41 . A targeted delivery composition comprising:
 a compound of  claim 26  wherein each of R 3  and R 4  is an attachment component attached to a nanoparticle, and R 1  is selected from a group consisting of a targeting agent, a diagnostic agent and a stealth agent,   or   a compound of formula:   
       
         
           
           
               
               
           
         
       
       wherein
 each L 1 , L 2 , L 3  and L 4  is independently selected from the group consisting of a bond and a linking group; 
 R 1  is selected from the group consisting of a targeting agent, a diagnostic agent and a stealth agent; 
 each R 2  is independently selected from the group consisting of a stealth agent, C 1 -C 10  alkyl, a carboxylic acid or ester, a phosphonic acid or ester, a sulfonic acid or ester, and a hydroxy; 
 each R 3  and R 4  is an attachment component attached to a nanoparticle; and 
 n is an integer from 0 to 2. 
 
     
     
         42 . A targeted delivery composition comprising:
 a compound of  claim 26 , wherein R 1  is a nanoparticle or an attachment component attached to a nanoparticle, and each of R 3  and R 4  is independently selected from a targeting agent, a diagnostic agent and a stealth agent,   or
 a compound of formula: 
   
       
         
           
           
               
               
           
         
       
       wherein
 each L 1 , L 2 , L 3  and L 4  is independently selected from the group consisting of a bond and a linking group; 
 R 1  is selected from the group consisting of a nanoparticle or an attachment component attached to a nanoparticle; 
 each R 2  is independently selected from the group consisting of a stealth agent, C 1 -C 10  alkyl, a carboxylic acid or ester, a phosphonic acid or ester, a sulfonic acid or ester, and a hydroxy; 
 each R 3  and R 4  is independently selected from a targeting agent, a diagnostic agent and a stealth agent; and
 n is an integer from 0 to 2. 
 
 
     
     
         43 . A method for treating or diagnosing a cancerous condition in a subject, comprising administering to the subject a targeted delivery composition of  claim 42 , wherein the composition comprises a therapeutic or a diagnostic agent that is sufficient to treat or diagnose the condition, and wherein at least one of R 1 , R 3  and R 4  is a targeting agent. 
     
     
         44 . A method of determining the suitability of a subject for a targeted therapeutic treatment, comprising administering to the subject a targeted delivery composition of  claim 42 , wherein at least one of R 1 , R 3  and R 4  is a targeting agent and the nanoparticle comprises a diagnostic agent, and imaging the subject to detect the diagnostic agent. 
     
     
         45 . The method of  claim 43 , wherein the nanoparticle is a liposome and the therapeutic agent is encapsulated in, embedded in, or tethered to the liposome. 
     
     
         46 . The method of  claim 45 , wherein the therapeutic agent is an anticancer agent selected from the group consisting of doxorubicin, cisplatin, oxaliplatin, carboplatin, 5-fluorouracil, gemcitibine and a taxane.