Mixtures of drug-oligomer conjugates comprising polyalkylene glycol, uses thereof, and methods of making same

Abstract

A non-polydispersed mixture of conjugates in which each conjugate in the mixture comprises a drug coupled to an oligomer that includes a polyalkylene glycol moiety is disclosed. The mixture may exhibit higher in vivo activity than a polydispersed mixture of similar conjugates. The mixture may be more effective at surviving an in vitro model of intestinal digestion than polydispersed mixtures of similar conjugates. The mixture may result in less inter-subject variability than polydispersed mixtures of similar conjugates.

1 . A substantially monodispersed mixture of conjugates, each conjugate comprising a drug coupled to at least one oligomer, wherein the oligomer comprises a polyethylene glycol moiety having at least 2 polyethylene glycol subunits, wherein at least 95% of the conjugates have the same molecule weight and wherein the substantially monodispersed mixture of conjugates has increased activity relative to a polydispersed mixture having the same average molecular weight.  
     
     
         2 . The substantially monodispersed mixture according to  claim 1 , wherein the polyethylene glycol moiety has at least 5 polyethylene glycol subunits.  
     
     
         3 . The substantially monodispersed mixture according to  claim 1 , wherein the polyethylene glycol moiety has at least 7 polyethylene glycol subunits.  
     
     
         4 . The substantially monodispersed mixture according to  claim 1 , wherein the oligomer is covalently coupled to the drug.  
     
     
         5 . The substantially monodispersed mixture according to  claim 1 , wherein the oligomer further comprises a lipophilic moiety.  
     
     
         6 . The substantially monodispersed mixture according to  claim 1 , wherein the oligomer is devoid of a lipophilic moiety, and wherein the conjugate is amphiphilically balanced such that it is aqueously soluble and able to penetrate biological membranes.  
     
     
         7 . The substantially monodispersed mixture according to  claim 1 , wherein the mixture is a monodispersed mixture.  
     
     
         8 . The substantially monodispersed mixture according to  claim 1 , wherein the mixture is a substantially purely monodispersed mixture.  
     
     
         9 . The substantially monodispersed mixture according to  claim 1 , wherein at least 96 percent of the conjugates in the mixture have the same molecular weight and the same molecular structure.  
     
     
         10 . The substantially monodispersed mixture according to  claim 1 , wherein the mixture is a purely monodispersed mixture.  
     
     
         11 . The substantially monodispersed mixture according to  claim 10 , wherein the oligomer is covalently coupled to the drug.  
     
     
         12 . The substantially monodispersed mixture according to  claim 10 , wherein the oligomer further comprises a lipophilic moiety.  
     
     
         13 . The substantially monodispersed mixture according to  claim 10 , wherein the polyethylene glycol moiety is uniform.  
     
     
         14 . The substantially monodispersed mixture according to  claim 13 , wherein the oligomer is devoid of a lipophilic moiety, and wherein the conjugate is amphiphilically balanced such that it is aqueously soluble and able to penetrate biological membranes.  
     
     
         15 . The substantially monodispersed mixture according to  claim 1 , wherein the mixture has an in vivo activity that is greater than the in vivo activity of a polydispersed mixture of drug-oligomer conjugates having the same number average molecular weight as the mixture.  
     
     
         16 . The substantially monodispersed mixture according to  claim 1 , wherein the mixture has an in vitro activity that is greater than the in vitro activity of a polydispersed mixture of drug-oligomer conjugates having the same number average molecular weight as the mixture.  
     
     
         17 . The substantially monodispersed mixture according to  claim 1 , wherein the mixture has an increased resistance to degradation by chymotrypsin when compared to the resistance to degradation by chymotrypsin of a polydispersed mixture of drug-oligomer conjugates having the same number average molecular weight as the mixture.  
     
     
         18 . The substantially monodispersed mixture according to  claim 1 , wherein the drug is a polypeptide.  
     
     
         19 . The substantially monodispersed mixture according to  claim 18 , wherein the polypeptide is selected from the group consisting of adrenocorticotropic hormone peptides, adrenomedullin peptides, allatostatin peptides, amylin peptides, amyloid beta-protein fragment peptides, angiotensin peptides, antibiotic peptides, antigenic polypeptides, anti-microbial peptides, apoptosis related peptides, atrial natriuretic peptides, bag cell peptides, bombesin peptides, bone GLA peptides, bradykinin peptides, brain natriuretic peptides, C-peptides, C-type natriuretic peptides, calcitonin peptides, calcitonin gene related peptides, CART peptides, casomorphin peptides, chemotactic peptides, cholecystokinin peptides, colony-stimulating factor peptides, corticortropin releasing factor peptides, cortistatin peptides, cytokine peptides, dermorphin peptides, dynorphin peptides, endorphin peptides, endothelin peptides, ETa receptor antagonist peptides, ETb receptor antagonist peptides, enkephalin peptides, fibronectin peptides, galanin peptides, gastrin peptides, glucagon peptides, Gn-RH associated peptides, growth factor peptides, growth hormone peptides, GTP-binding protein fragment peptides, guanylin peptides, inhibin peptides, insulin peptides, interleukin peptides, laminin peptides, leptin peptides, leucokinin peptides, luteinizing hormone-releasing hormone peptides, mastoparan peptides, mast cell degranulating peptides, melanocyte stimulating hormone peptides, morphiceptin peptides, motilin peptides, neuro-peptides, neuropeptide Y peptides, neurotropic factor peptides, orexin peptides, opioid peptides, oxytocin peptides, PACAP peptides, pancreastatin peptides, pancreatic polypeptides, parathyroid hormone peptides, parathyroid hormone-related peptides, peptide T peptides, prolactin-releasing peptides, peptide YY peptides, renin substrate peptides, secretin peptides, somatostatin peptides, substance P peptides, tachykinin peptides, thyrotropin-releasing hormone peptides, toxin peptides, vasoactive intestinal peptides, vasopressin peptides, and virus related peptides.  
     
     
         20 . The substantially monodispersed mixture according to  claim 18 , wherein the oligomer is covalently coupled to a nucleophilic residue of the polypeptide.  
     
     
         21 . The substantially monodispersed mixture according to  claim 1 , wherein each conjugate comprises a plurality of oligomers.  
     
     
         22 . The substantially monodispersed mixture according to  claim 21 , wherein each oligomer in the plurality of oligomers is the same.  
     
     
         23 . The substantially monodispersed mixture according to  claim 1 , wherein the oligomer comprises a first polyethylene glycol moiety covalently coupled to the drug by a non-hydrolyzable bond and a second polyethylene glycol moiety covalently coupled to the first polyethylene glycol moiety by a hydrolyzable bond.  
     
     
         24 . The substantially monodispersed mixture according to  claim 23 , wherein the oligomer further comprises a lipophilic moiety covalently coupled to the second polyethylene glycol moiety.  
     
     
         25 . A pharmaceutical composition comprising: 
 the mixture according to  claim 1;  and    a pharmaceutically acceptable carrier.    
     
     
         26 . A substantially monodispersed mixture of conjugates, each conjugate comprising a drug coupled to an oligomer that comprises a polyethylene glycol moiety and wherein the oligomer is devoid of a lipophilic moiety, wherein at least 95% of the conjugates have the same molecule weight and wherein the conjugate is amphiphilically balanced such that it is aqueously soluble and able to penetrate biological membranes.  
     
     
         27 . The substantially monodispersed mixture according to  claim 26 , wherein the polyethylene glycol moiety is uniform.  
     
     
         28 . The substantially monodispersed mixture according to  claim 26 , wherein the oligomer is covalently coupled to the drug.  
     
     
         29 . A substantially monodispersed mixture of conjugates each comprising a drug coupled to an oligomer that comprises a polyethylene glycol moiety, said mixture having a molecular weight distribution with a standard deviation of less than about 22 Daltons.  
     
     
         30 . The substantially monodispersed mixture according to  claim 29 , wherein the standard deviation of the molecular weight distribution is less than about 14 Daltons.  
     
     
         31 . The substantially monodispersed mixture according to  claim 29 , wherein the polyethylene glycol moiety has at least 7 polyethylene glycol subunits.  
     
     
         32 . The substantially monodispersed mixture according to  claim 29 , wherein the oligomer further comprises a lipophilic moiety.  
     
     
         33 . The substantially monodispersed mixture according to  claim 29 , wherein the oligomer is devoid of a lipophilic moiety, and wherein the conjugate is amphiphilically balanced such that it is aqueously soluble and able to penetrate biological membranes.  
     
     
         34 . The substantially monodispersed mixture according to  claim 29 , wherein the drug is a polypeptide selected from the group consisting of adrenocorticotropic hormone peptides, adrenomedullin peptides, allatostatin peptides, amylin peptides, amyloid beta-protein fragment peptides, angiotensin peptides, antibiotic peptides, antigenic polypeptides, anti-microbial peptides, apoptosis related peptides, atrial natriuretic peptides, bag cell peptides, bombesin peptides, bone GLA peptides, bradykinin peptides, brain natriuretic peptides, C-peptides, C-type natriuretic peptides, calcitonin peptides, calcitonin gene related peptides, CART peptides, casomorphin peptides, chemotactic peptides, cholecystokinin peptides, colony-stimulating factor peptides, corticortropin releasing factor peptides, cortistatin peptides, cytokine peptides, dermorphin peptides, dynorphin peptides, endorphin peptides, endothelin peptides, ETa receptor antagonist peptides, ETb receptor antagonist peptides, enkephalin peptides, fibronectin peptides, galanin peptides, gastrin peptides, glucagon peptides, Gn-RH associated peptides, growth factor peptides, growth hormone peptides, GTP-binding protein fragment peptides, guanylin peptides, inhibin peptides, insulin peptides, interleukin peptides, laminin peptides, leptin peptides, leucokinin peptides, luteinizing hormone-releasing hormone peptides, mastoparan peptides, mast cell degranulating peptides, melanocyte stimulating hormone peptides, morphiceptin peptides, motilin peptides, neuro-peptides, neuropeptide Y peptides, neurotropic factor peptides, orexin peptides, opioid peptides, oxytocin peptides, PACAP peptides, pancreastatin peptides, pancreatic polypeptides, parathyroid hormone peptides, parathyroid hormone-related peptides, peptide T peptides, prolactin-releasing peptides, peptide YY peptides, renin substrate peptides, secretin peptides, somatostatin peptides, substance P peptides, tachykinin peptides, thyrotropin-releasing hormone peptides, toxin peptides, vasoactive intestinal peptides, vasopressin peptides, and virus related peptides.  
     
     
         35 . A substantially monodispersed mixture of conjugates each comprising a drug coupled to a polymer comprising a polyethylene glycol moiety, wherein the mixture has a dispersity coefficient (DC) greater than 10,000 where  
       
         
           
             
               
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       wherein: 
 n is the number of different molecules in the sample;  
 N i  is the number of i th  molecules in the sample; and  
 M i  is the mass of the i th  molecule.  
 
     
     
         36 . The substantially monodispersed mixture according to  claim 35 , wherein the dispersity coefficient is greater than 100,000.  
     
     
         37 . The substantially monodispersed mixture according to  claim 35 , wherein the polyethylene glycol moiety has at least 7 polyethylene glycol subunits.  
     
     
         38 . The substantially monodispersed mixture according to  claim 35 , wherein the polymer further comprises a lipophilic moiety.  
     
     
         39 . The substantially monodispersed mixture according to  claim 35 , wherein the polyethylene glycol moiety is uniform.  
     
     
         40 . The substantially monodispersed mixture according to  claim 35 , wherein the conjugates have the same number of polyethylene glycol subunits.  
     
     
         41 . A substantially monodisperse mixture of conjugates in which each conjugate has the same molecular weight and has the formula:  
         Drug B-L h -G i -R′ n -G″ k -T] p   (A)  
       wherein: 
 B is a bonding moiety;  
 L is a linker moiety;  
 G, G′ and G″ are individually selected spacer moieties;  
 R is a lipophilic moiety and R′ is a polypropylene glycol moiety, or R′ is a lipophilic moiety and R is a polyethylene glycol moiety;  
 T is a terminating moiety;  
 h, i, j, k, m and n are individually 0 or 1, with the proviso that when R is a polyethylene glycol moiety, m is 1; and when R′ is a polyethylene glycol moiety, n is 1; and  
 p is an integer from 1 to the number of nucleophilic residues on the drug.  
 
     
     
         42 . The substantially monodispersed mixture according to  claim 41 , wherein the polyethylene glycol moiety has at least 7 polyethylene glycol subunits.  
     
     
         43 . The substantially monodispersed mixture according to  claim 41 , wherein: 
 R is the polyethylene glycol moiety;    R′ is a lipophilic moiety;    n and m are 1; and    i, j and k are 0.    
     
     
         44 . The substantially monodispersed mixture according to  claim 41 , wherein: 
 R is a lipophilic moiety;    R′ is the polyethylene glycol moiety;    n and m are 1; and    i, j and k are each 0.    
     
     
         45 . The substantially monodispersed mixture according to  claim 41 , wherein the polyethylene glycol moiety is uniform.  
     
     
         46 . The substantially monodispersed mixture according to  claim 41 , wherein: 
 R is the polyethylene glycol moiety;    m is 1;    i, j, k and n are each 0; and    each conjugate in the mixture is amphiphilically balanced such that each conjugate is aqueously soluble and able to penetrate biological membranes.    
     
     
         47 . The substantially monodispersed mixture according to  claim 41 , wherein the drug is a polypeptide selected from the group consisting of adrenocorticotropic hormone peptides, adrenomedullin peptides, allatostatin peptides, amylin peptides, amyloid beta-protein fragment peptides, angiotensin peptides, antibiotic peptides, antigenic polypeptides, anti-microbial peptides, apoptosis related peptides, atrial natriuretic peptides, bag cell peptides, bombesin peptides, bone GLA peptides, bradykinin peptides, brain natriuretic peptides, C-peptides, C-type natriuretic peptides, calcitonin peptides, calcitonin gene related peptides, CART peptides, casomorphin peptides, chemotactic peptides, cholecystokinin peptides, colony-stimulating factor peptides, corticortropin releasing factor peptides, cortistatin peptides, cytokine peptides, dermorphin peptides, dynorphin peptides, endorphin peptides, endothelin peptides, ETa receptor antagonist peptides, ETb receptor antagonist peptides, enkephalin peptides, fibronectin peptides, galanin peptides, gastrin peptides, glucagon peptides, Gn-RH associated peptides, growth factor peptides, growth hormone peptides, GTP-binding protein fragment peptides, guanylin peptides, inhibin peptides, insulin peptides, interleukin peptides, laminin peptides, leptin peptides, leucokinin peptides, luteinizing hormone-releasing hormone peptides, mastoparan peptides, mast cell degranulating peptides, melanocyte stimulating hormone peptides, morphiceptin peptides, motilin peptides, neuro-peptides, neuropeptide Y peptides, neurotropic factor peptides, orexin peptides, opioid peptides, oxytocin peptides, PACAP peptides, pancreastatin peptides, pancreatic polypeptides, parathyroid hormone peptides, parathyroid hormone-related peptides, peptide T peptides, prolactin-releasing peptides, peptide YY peptides, renin substrate peptides, secretin peptides, somatostatin peptides, substance P peptides, tachykinin peptides, thyrotropin-releasing hormone peptides, toxin peptides, vasoactive intestinal peptides, vasopressin peptides, and virus related peptides.  
     
     
         48 . A pharmaceutical composition comprising: 
 the substantially monodispersed mixture according to  claim 10;  and    pharmaceutically acceptable carrier.    
     
     
         49 . A pharmaceutical composition comprising: 
 the substantially monodispersed mixture according to  claim 26;  and    a pharmaceutically acceptable carrier.    
     
     
         50 . A pharmaceutical composition comprising; 
 the substantially monodispersed mixture according to  claim 29;  and    a pharmaceutically acceptable carrier.    
     
     
         51 . A pharmaceutical composition comprising: 
 the substantially monodispersed mixture according to  claim 41;  and    a pharmaceutically acceptable carrier.    
     
     
         52 . A pharmaceutical composition comprising: 
 the substantially monodispersed mixture according to  claim 41;  and    a pharmaceutically acceptable carrier.    
     
     
         53 . A process for synthesizing a substantially monodispersed mixture of conjugates each conjugate comprising a drug coupled to an oligomer that comprises a polyethylene glycol moiety, said process comprising: 
 reacting a substantially monodispersed mixture comprising compounds having the structure of Formula I:      R 1 (OC 2 H 4 ) m —O − X +   (1)  wherein R 1  is H or a lipophilic moiety; m is from 1 to 25; and X +  is a positive ion, with a substantially monodispersed mixture comprising compounds having the structure of Formula II:      R 2 (OC 2 H 4 ) n —OMs  (II)      wherein R 2  is a fatty acid moiety or an ester of a fatty acid moiety; and n is from 1 to 25, under conditions sufficient to provide a substantially monodispersed mixture comprising polymers having the structure of Formula III:      R 2 (OC 2 H 4 ) m+n —OR 1   (III);    activating the substantially monodispersed mixture comprising polymers of Formula III to provide a substantially monodispersed mixture of activated polymers capable of reacting with a drug; and    reacting the substantially monodispersed mixture of activated polymers with a drug under conditions sufficient to provide a substantially monodispersed mixture of conjugates each comprising a drug coupled to an oligomer that comprises a polyethylene glycol moiety with m+n subunits.    
     
     
         54 . The process according to  claim 53 , wherein the fatty acid moiety or the ester of a fatty acid moiety comprises an alkyl moiety at least 5 carbon atoms in length.  
     
     
         55 . The process according to  claim 53 , wherein R 1  is a methyl group.  
     
     
         56 . The process according to  claim 53 , further comprising: 
 reacting a substantially monodispersed mixture comprising compounds having the structure of Formula V:      R 2 (OC 2 H 4 ) n —OH  (V)    with a methanesulfonyl halide under conditions sufficient to provide a substantially monodispersed mixture comprising compounds having the structure of Formula II:      R 2 (OC 2 H 4 ) n —OMs  (II).    
     
     
         57 . A process for synthesizing a substantially monodispersed mixture of conjugates each conjugate comprising a drug coupled to an oligomer that comprises a polyethylene glycol moiety, said process comprising: 
 reacting a substantially monodispersed mixture comprising compounds having the structure of Formula VI:      R 2 —OMs  (VI)    wherein R 2  is a lipophilic moiety, with a substantially monodispersed mixture comprising compounds having the structure of Formula VII:      R 3 (OC 2 H 4 ) m —O − X 2   +   (VII)    wherein R 3  is benzyl, trityl, or THP; and X 2   +  is a positive ion, under conditions sufficient to provide a substantially monodispersed mixture comprising compounds having the structure of Formula VIII:      R 3 (OC 2 H 4 ) m —OR 2   (VIII);    reacting the substantially monodispersed mixture comprising compounds having the structure of Formula VIII, under conditions sufficient to provide a substantially monodispersed mixture comprising compounds having the structure of Formula V:      R 2 (OC 2 H 4 ) m —OH  (V);    reacting the substantially monodispersed mixture comprising compounds having the structure of Formula V, with a methanesulfonyl halide comprising compounds having the structure CH 3 SO 2 Q, wherein Q is a halide, under conditions sufficient to provide a substantially monodispersed mixture comprising compounds having the structure of Formula II:      R 2 (OC 2 H 4 ) n —OMs  (II)    wherein R 2  is a lipophilic moiety; and n is from 1 to 25;    reacting a substantially monodispersed mixture comprising compounds having the structure of Formula I:      R 1 (OC 2 H 4 ) m —O − X +   (I)    wherein R 1  is H or a lipophilic moiety; m is from 1 to 25; and X +  is a positive ion, with the substantially monodispersed mixture comprising compounds having the structure of Formula II, under conditions sufficient to provide a substantially monodispersed mixture comprising polymers having the structure of Formula III:      R 2 (OC 2 H 4 ) m+n —OR 1   (III);    activating the substantially monodispersed mixture comprising polymers of Formula III to provide a substantially monodispersed mixture of activated polymers capable of reacting with a drug; and    reacting the substantially monodispersed mixture of activated polymers with a drug under conditions sufficient to provide a substantially monodispersed mixture of conjugates each comprising a drug coupled to an oligomer that comprises a polyethylene glycol moiety with m+n subunits.    
     
     
         58 . The process according to  claim 57 , further comprising: 
 reacting a substantially monodispersed mixture comprising compounds having the structure of Formula IV:      R 1 (OC 2 H 4 ) n —OH  (IV)    under conditions sufficient to provide a substantially monodispersed mixture comprising compounds having the structure of Formula I:      R 1 (OC 2 H 4 ) n —O − X +   (I).    
     
     
         59 . The process according to  claim 57 , wherein the activating of the substantially monodispersed mixture comprises reacting the substantially monodispersed mixture of polymers of Formula III with N-hydroxy succinimide to provide an activated polymer capable of reacting with a drug.  
     
     
         60 . The process according to claim  159 , wherein the drug is a polypeptide, and wherein the reacting of the substantially monodispersed mixture of activated polymers with a substantially monodispersed mixture of polypeptides comprises: 
 reacting the substantially monodispersed mixture of activated polymers with one or more amino functionalities of the polypeptide to provide a substantially monodispersed mixture of conjugates each comprising the polypeptide coupled to an oligomer that comprises a polyethylene glycol moiety with m+n subunits.