Method for Preparing Nanoparticles Based on Functional Amphiphilic Molecules or Macromolecules, and the Use Thereof

Abstract

The invention relates to a method for preparing nanoparticles based on functional amphiphilic molecules or macromolecules, optionally in the presence of at least one colipide, enabling the encapsulation of therapeutic agents, especially anti-tumoral agents, and the use thereof for the transport and vectorisation of therapeutic agents, especially anti-tumoral agents.

1 . Process for encapsulating a therapeutic agent comprising the stages consisting of:
 a) preparing a mixture of at least one functional amphiphilic compound of formula (I)   
       
         
           
           
               
               
           
         
       
       in which
 X represents an oxygen or sulphur atom or a methylene group, 
 B represents a purine or pyrimidine base such as uracil, adenine, guanine, cytosine, thymine, hypoxanthine, or their derivatives, or also a non-natural mono- or bi-cyclic heterocyclic base each ring of which comprises 4 to 7 members, optionally substituted; 
 L 1  and L 2 , identical or different, represent hydrogen, an oxycarbonyl —O—C(O)— group, a thiocarbamate —O—C(S)—NH— group, a carbonate —O—C(O)—O— group, a carbamate —O—C(O)—NH— group, an oxygen atom, a phosphate group, a phosphonate group or a heteroaryl group comprising 1 to 4 nitrogen atoms, unsubstituted or substituted by a linear or branched, saturated or unsaturated C 2 -C 30  hydrocarbon chain, 
 or also, L 1  and L 2 , together, form a ketal group of formula 
 
       
         
           
           
               
               
           
         
         or also L 1  or L 2  represents hydrogen, and the other represents a hydroxy group or a heteroaryl group comprising 1 to 4 nitrogen atoms, unsubstituted or substituted by a linear or branched C 2 -C 30  alkyl chain; 
         R 1  and R 2 , identical or different, represent
 a linear or branched C 2 -C 30  hydrocarbon chain, preferably C 6 -C 25 , in particular C 8 -C 25 , saturated or partially unsaturated, optionally completely or partially fluorinated, unsubstituted or substituted on the carbon at the end of the chain by a fluorine atom or by a benzyl or naphthyl ester or ether, or 
 a diacyl chain in which each acyl chain is C 2 -C 30 , or 
 a diacylglycerol, sphingosine or ceramide group, or 
 when L 1  or L 2  represents hydrogen, and the other represents a hydroxy group or a heteroaryl group comprising 1 to 4 nitrogen atoms, R 1  and R 2  do not exist; 
 
         R 3  represents a hydroxy, amino, phosphate, phosphonate, phosphatidylcholine, O-alkyl phosphatidylcholine, thiophosphate, phosphonium, NH 2 —R 4 , NHR 4 R 5  or NR 4 R 5 R 6  group in which R 4 , R 5  and R 6 , identical or different, represent a hydrogen atom or a linear or branched C 1 -C 5  alkyl or hydroxyalkyl chain, or
 a linear or branched C 2 -C 30  alkyl chain optionally substituted by a hydroxy group, or 
 a cyclodextrin radical, or 
 a 
 
       
       
         
           
           
               
               
           
         
         radical, in which V represents an —O—, —S—, or —NH— bond, R 7  represents H or CH 3 , and n=1 to 500, or
 a —(CH 2 ) n —V—R 8  group, in which R 8  represents a C 2 -C 30  alkyl, and n=1 to 500, or 
 a heteroaryl group containing 1 to 4 nitrogen atoms, unsubstituted or substituted by a C 2 -C 30  alkyl, or by a (CH 2 ) m —O—(CH 2 ) p —R 9  group in which m=1 to 6 and p=0 to 10 and R 9  represents a cyclic ketal group containing 5 to 7 carbon atoms, unsubstituted or substituted by at least one linear or branched C 2 -C 30  alkyl or by a sterol radical, or also 
 
         R 3  is bound by a covalent bond to another substituent R 3 , identical or different, of another compound of formula (I), identical or different, in order to form a compound of formula (I) in the form of a dimer, and 
         a therapeutic agent, preferably an anti-neoplastic agent, 
         b) subjecting said mixture to repeated heating and freezing cycles, in order to obtain nanoparticles containing said therapeutic agent, and 
         c) recovering the nanoparticles containing said therapeutic agent obtained in this way. 
       
     
     
         2 . Process according to  claim 1 , characterized in that, in formula (I), X represents oxygen. 
     
     
         3 . Process  claim 1 , wherein, in formula (I), B represents thymine or adenine. 
     
     
         4 . Process according to  claim 1 , wherein, in formula (I).
 X and B are as defined in  claim 1 ;   L 1  represents a phosphate group, L 2  represents hydrogen, R 1  represents a C 2 -C 30  alkyl group or a diacyl group in which each acyl chain is C 2 -C 30  and R 3  is a hydroxy group; or   L 1  and L 2  represent an oxygen atom, R 1  and R 2  represent hydrogen and R 3  represents a triazole, tetrazole, pyrazole or imidazole group substituted by a C 2 -C 30  alkyl group, or   L 1  represents a triazole, tetrazole, pyrazole or imidazole group, L 2  represents hydrogen, R 1  represents a C 2 -C 30  alkyl group, and R 3  is a hydroxy group, or   L 1  represents a hydroxy group, L 2  represents hydrogen and R 3  is a triazole group substituted by a C 2 -C 30  alkyl chain, optionally substituted by a hydroxy group, or R 3  is a group or a —(CH 2 )n-V—R 8  group, in which V represents —O— or —NH— and R 8  represents a C 2 -C 30  alkyl, or   L 1  represents a hydroxy group, L 2  represents hydrogen and R 3  is a triazole group substituted by a (CH 2 ) m —O—(CH 2 ) p —R 9  group in which m=1 to 6 and p=0 to 10 and R 9  represents a cyclic ketal group containing 5 to 7 carbon atoms, unsubstituted or substituted by at least one linear or branched C 2 -C 30  alkyl group or by a sterol radical, or   L 1  represents a hydroxy group, L 2  represents hydrogen and R 3  is a triazole group substituted by a (CH 2 ) m —O—(CH 2 ) p —R 9  group in which m=1 to 6 and p=0 to 10 bound by a covalent bond to another identical substituent R 3 , of another identical compound of formula (I), in order to form a compound of formula (I) in the form of a dimer, or   L 1  represents a phosphate group, L 2  represents hydrogen, R 1  represents a diacyl chain in which each diacyl chain is C 2 -C 30 , and R 3  is a hydroxy group.   
     
     
         5 . Process according to  claim 1 , wherein in formula (I):
 X, B, L 2 , R 1  and R 2  are as defined in formula (I), and   the substituent L 1  or the substituent R 3  is constituted by or comprises a triazole group, unsubstituted or substituted.   
     
     
         6 . Process according to  claim 1 , further comprising the steps consisting of:
 putting the compound of formula (I) in solution in an organic solvent in order to form a lipid mixture,   evaporating said mixture in order to form a film,   putting the desired quantity of therapeutic agent, preferably an anti-neoplastic agent in solution in distilled water,   rehydrating the lipid film in the solution of therapeutic agent, preferably an anti-neoplastic agent,   subjecting the mixture obtained in this way to a heating and cooling cycle carried out 1 to 10 times.   
     
     
         7 . Process according to  claim 1 , further comprising the step of putting the compound of formula (I) in solution in an organic solvent in the presence of a co-lipid. 
     
     
         8 . Process according to  claim 1 , further comprising the step of putting the compound of formula (I) t in solution in an organic solvent in the presence of a co-lipid, wherein the co-lipid is chosen from dioleylphosphatidylcholine (DOPC) and dioleyl phosphatidyluridinephosphatidylcholine (DOUPC). 
     
     
         9 . Process according to  claim 1 , wherein the therapeutic agent is chosen from the platinum complexes or ruthenium capable of binding to platinum complexes, or also the inorganic complexes without platinum based on ruthenium II or III, titanium, gallium, cobalt, iron or gold. 
     
     
         10 . Process according to  claim 1 , wherein the therapeutic agent is chosen from cis-platin, carboplatin, oxaliplatin, nedaplatin and lobaplatin. 
     
     
         11 . Nanoparticles capable of being obtained by the process according to  claim 1 . 
     
     
         12 . Solid nanoparticles, wherein they are constituted by a core rich in therapeutic agent, preferably an anti-neoplastic agent, surrounded by one or more lipid layers constituted by the compound of formula (I), 
       
         
           
           
               
               
           
         
       
       in which
 X represents an oxygen or sulphur atom, or a methylene group, 
 B represents a purine or pyrimidine base such as uracil, adenine, guanine, cytosine, thymine, hypoxanthine, or their derivatives, or also a non-natural mono- or bi-cyclic heterocyclic base each ring of which comprises 4 to 7 members, optionally substituted; 
 L 1  and L 2 , identical or different, represent hydrogen, an oxycarbonyl —O—C(O)— group, a thiocarbamate —O—C(S)—NH— group, a carbonate —O—C(O)—O— group, a carbamate —O—C(O)—NH— group, an oxygen atom, a phosphate group, a phosphonate group or a heteroaryl group comprising 1 to 4 nitrogen atoms, unsubstituted or substituted by a linear or branched, saturated or unsaturated, C 2 -C 30  hydrocarbon chain, 
 or also, L 1  and L 2 , together, form a ketal group of formula 
 
       
         
           
           
               
               
           
         
         or also L 1  or L 2  represents hydrogen, and the other represents a hydroxy group or a heteroaryl group comprising 1 to 4 nitrogen atoms, unsubstituted or substituted by a linear or branched C 2 -C 30  alkyl chain; 
         R 1  and R 2 , identical or different, represent
 a linear or branched C 2 -C 30  hydrocarbon chain, preferably C 6 -C 25 , in particular C 8 -C 25 , saturated or partially unsaturated, optionally completely or partially fluorinated, unsubstituted or substituted on the carbon at the end of the chain by a fluorine atom or by a benzyl or naphthyl ester or ether, or 
 a diacyl chain in which each acyl chain is C 2 -C 30 , or 
 a diacylglycerol, sphingosine or ceramide group, or 
 when L 1  or L 2  represents hydrogen, and the other represents a hydroxy group or a heteroaryl group comprising 1 to 4 nitrogen atoms, R 1  and R 2  do not exist; 
 
         R 3  represents a hydroxy, amino, phosphate, phosphonate, phosphatidylcholine, O-alkyl phosphatidylcholine, thiophosphate, phosphonium, NH 2 —R 4 , NHR 4 R 5  or NR 4 R 5 R 6  group in which R 4 , R 5  and R 6 , identical or different, represent a hydrogen atom or a linear or branched C 1 -C 5  alkyl or hydroxyalkyl chain, or
 a linear or branched C 2 -C 30  alkyl chain optionally substituted by a hydroxy group, or 
 a cyclodextrin radical, or 
 a 
 
       
       
         
           
           
               
               
           
         
         radical, in which V represents an —O—, —S—, or —NH— bond, R 7  represents H or CH 3 , and n=1 to 500, or
 a —(CH 2 ) n —V—R 8  group, in which R 8  represents a C 2 -C 30  alkyl, and n=1 to 500, or 
 a heteroaryl group containing 1 to 4 nitrogen atoms, unsubstituted or substituted by a C 2 -C 30  alkyl, or by a (CH 2 ) m —O—(CH 2 ) p —R 9  group in which m=1 to 6 and p=0 to 10 and R 9  represents a cyclic ketal group containing 5 to 7 carbon atoms, unsubstituted or substituted by at least one linear or branched C 2 -C 30  alkyl or by a sterol radical, or also
 R 3  is bound by a covalent bond to another substituent R 3 , identical or different, of another compound of formula (I), identical or different, in order to form a compound of formula (I) in the form of a dimer, 
 with or without a co-lipid. 
 
 
       
     
     
         13 . Compound of formula (I): 
       
         
           
           
               
               
           
         
         in which
 X represents an oxygen or sulphur atom, 
 B represents a purine or pyrimidine base such as uracil, adenine, guanine, cytosine, thymine, hypoxanthine, or their derivatives, or also a non-natural mono- or bi-cyclic heterocyclic base each ring of which comprises 4 to 7 members, optionally substituted; and L 1 , L 2 , R 1 , R 2  and R 3  are chosen from one of the following definitions: 
 
         L 1  represents a phosphate group, L 2  represents hydrogen, R 1  represents a C 2 -C 30  alkyl group or a diacyl group in which each acyl chain is C 2 -C 30 , and R 3  is a hydroxy group; or 
         L 1  and L 2  represent an oxygen atom, R 1  and R 2  represent hydrogen and R 3  represents a triazole, tetrazole, pyrazole or imidazole group substituted by a C 2 -C 30  alkyl group, or 
         L 1  represents a triazole, tetrazole, pyrazole or imidazole group, L 2  represents hydrogen, R 1  represents a C 2 -C 30  alkyl group, and R 3  is a hydroxy group, or 
         L 1  represents a hydroxy group, L 2  represents hydrogen and R 3  is a triazole group substituted by a C 2 -C 30  alkyl chain, optionally substituted by a hydroxy group, or R 3  is a group or a —(CH 2 )n-V—R 8  group, in which V represents —O— or —NH— and R 8  represents a C 2 -C 30  alkyl, or 
         L 1  represents a hydroxy group, L 2  represents hydrogen and R 3  is a triazole group substituted by a (CH 2 ) m —O—(CH 2 ) p —R 9  group in which m=1 to 6 and p=0 to 10 and R 9  represents a cyclic ketal group containing 5 to 7 carbon atoms, unsubstituted or substituted by at least one linear or branched C 2 -C 30  alkyl group or by a sterol radical, or 
         L 1  represents a hydroxy group, L 2  represents hydrogen and R 3  is a triazole group substituted by a (CH 2 ) m —O—(CH 2 ) p —R 9  group in which m=1 to 6 and p=0 to 10 bound by a covalent bond to another identical substituent R 3  of another identical compound of formula (I), in order to form a compound of formula (I) in the form of a dimer, or 
         L 1  represents a phosphate group, L 2  represents hydrogen, R 1  represents a diacyl chain in which each diacyl chain is C 2 -C 30 , and R 3  is a hydroxy group. 
       
     
     
         14 . Compound of formula (I): 
       
         
           
           
               
               
           
         
         in which
 X, B, L 2  and R 2  are as defined in  claim 1 ; 
 L 1  or R 3  is constituted by or comprises a triazole group, unsubstituted or substituted. 
 
       
     
     
         15 . Nanoparticle comprising a compound of formula (I) according to  claim 13  and a therapeutic agent. 
     
     
         16 . Nanoparticle comprising a compound of formula (I) according to  claim 13  and a therapeutic agent, wherein the therapeutic agent is an anti-neoplastic agent. 
     
     
         17 . Nanoparticle according to  claim 11 , wherein the therapeutic agent is chosen from cis-platin, carboplatin, oxaliplatin, nedaplatin and lobaplatin. 
     
     
         18 . (canceled) 
     
     
         19 . (canceled) 
     
     
         20 . (canceled) 
     
     
         21 . (canceled) 
     
     
         22 . Process according to  claim 1 , wherein in formula (I) X represents oxygen and B represents thymine or adenine. 
     
     
         23 . Nanoparticle according to  claim 12 , wherein the therapeutic agent from is selected from the group consisting of cis-platin, carboplatin, oxaliplatin, nedaplatin and lobaplatin. 
     
     
         24 . Nanoparticle comprising a compound of formula (I) according to  claim 13  and a therapeutic agent, wherein the therapeutic agent is an anti-neoplastic agent. 
     
     
         25 . Nanoparticle comprising a compound of formula (I) according to  claim 13  and a therapeutic agent, wherein the therapeutic agent is selected from the group consisting of cis-platin, carboplatin, oxaliplatin, nedaplatin and lobaplatin. 
     
     
         26 . A method for transportation, vectorization or intracellular delivery of therapeutic agents, in particular of anti-neoplastic agents, wherein the nanoparticles according to  claim 11  are used as for transportation, vectorization or delivery agents. 
     
     
         27 . A method for transportation or vectorization of therapeutic agents, in particular of anti-neoplastic agents, wherein the nanoparticles according to  claim 12  are used as for transportation or vectorization agents. 
     
     
         28 . A method for transportation or vectorization of therapeutic agents, in particular of anti-neoplastic agents, wherein the nanoparticles according to  claim 13  are used as for transportation or vectorization agents. 
     
     
         29 . A method for transportation or vectorization of therapeutic agents, wherein the nanoparticles according to  claim 11  are used as for transportation or vectorization agents and the therapeutic agent is selected from the group consisting of cis-platin, carboplatin, oxaliplatin, nedaplatin and lobaplatin. 
     
     
         29 . A method of treating tumour diseases, wherein the nanoparticles according to  claim 11  are administered to a patient in need thereof. 
     
     
         30 . A method of treating tumour diseases, wherein the nanoparticles comprising a compound of formula (I) according to  claim 13  and an anti-neoplastic agent are administered to a patient in need thereof. 
     
     
         31 . A method of treating tumour diseases, wherein the nanoparticles according to  claim 11  wherein the therapeutic agent is selected from the group consisting of cis-platin, carboplatin, oxaliplatin, nedaplatin and lobaplatin. are administered to a patient in need thereof.