Enzymatic synthesis of poly(amine-co-esters) and methods of use thereof for gene delivery

Abstract

Poly(amine-co-ester) polymers, methods of forming active agent-load nanoparticles therefrom, and methods of using the nanoparticles for drug delivery are disclosed. The nanoparticles can be coated with an agent that reduces surface charge, an agent that increases cell-specific targeting, or a combination thereof. Typically, the loaded nanoparticles are less toxic, more efficient at drug delivery, or a combination thereof compared to a control other transfection reagents. In some embodiments, the nanoparticles are suitable for in vivo delivery, and can be administered systemically to a subject to treat a disease or condition.

We claim: 
     
         1 .- 44 . (canceled) 
     
     
         45 . An activated polymer comprising a backbone ester prepared by a process comprising exposing the polymer to conditions such that one or more backbone esters are hydrolyzed, thereby exposing one or more activated end group(s). 
     
     
         46 . The activated polymer of  claim 45 , wherein the polymer is a poly(amine-co-ester). 
     
     
         47 . The activated polymer of  claim 45 , wherein the one or more activated end group(s) are hydroxyl or carboxylic acid end groups. 
     
     
         48 . The activated polymer of  claim 45 , wherein the conditions for hydrolyzing one or more backbone esters comprise hydrolyzing one or more backbone esters of the polymer for about 1 day to about 30 days or more at a temperature from about 30° C. to 42° C. under atmospheric pressure. 
     
     
         49 . The activated polymer of  claim 46 , wherein the polymer is synthesized by a method comprising combining 15-pentadecanolide (PDL), diethanolamine, and a diester/diacids selected from either diethyl sebacate (DES) or sebacic acid (SBA). 
     
     
         50 . The activated polymer of  claim 49 , wherein the method is performed in the presence of a catalyst. 
     
     
         51 . The activated polymer of  claim 49 , wherein the method is performed at about 90° C. for about 24 hours. 
     
     
         52 . The activated polymer of  claim 49 , wherein the conditions for hydrolyzing one or more backbone esters comprise hydrolyzing one or more backbone esters of the polymer for about 1 day to about 30 days or more at a temperature from about 30° C. to about 42° C. under atmospheric pressure. 
     
     
         53 . The activated polymer of  claim 46 , wherein the activated polymer has a molecular weight of less than about 25 kDA. 
     
     
         54 . The activated polymer of  claim 53 , wherein the activated polymer has a molecular weight of less than about 15 kDa. 
     
     
         55 . The activated polymer of  claim 54 , wherein the activated polymer has a molecular weight of less than 10 kDa. 
     
     
         56 . A microparticle, nanoparticle or combination thereof comprising the activated polymer of  claim 45  and one or more therapeutic, prophylactic or diagnostic agents. 
     
     
         57 . The microparticle, nanoparticle or combination thereof of  claim 56 , wherein the agent is a macromolecule or small molecule. 
     
     
         58 . The microparticle, nanoparticle or combination thereof of  claim 57 , wherein the macromolecule is a polynucleotide. 
     
     
         59 . The microparticle, nanoparticle or combination thereof of  claim 58 , wherein the polynucleotide is mRNA. 
     
     
         60 . A method for activating a polymer comprising a backbone ester to produce a polymer suitable for delivery of an active pharmaceutical ingredient, comprising hydrolyzing one or more of the backbone esters of the polymer for about 1 day to about 30 days or more at a temperature from about 30° C. to 42° C. under atmospheric pressure. 
     
     
         61 . A method of making an activated poly(amine-co-ester) polymer, comprising:
 a. combining 15-pentadecanolide (PDL), diethanolamine, and a diester/diacids selected from either diethyl sebacate (DES) or sebacic acid (SBA) in the presence of a catalyst under atmospheric pressure at about 90° C. for 24 hours;   b. reducing the reaction pressure to 1.6 mmHg and continuing the reaction at about 90° C. for an additional 8 to 72 hours; and   c. hydrolyzing the terpolymers produced in step b) for about 1 day to about 30 days or more.   
     
     
         62 . A method of administering a macromolecule in vivo comprising administering the macromolecule formulated in a particle comprising an activated polymer comprising one or more hydrolysed backbone esters. 
     
     
         63 . The method of  claim 62 , wherein the activated polymer is an activated poly(amine-co-ester). 
     
     
         65 . The method of claim  64 , wherein the macromolecule is mRNA. 
     
     
         66 . The method of  claim 62 , wherein the macromolecular formulation further comprises a pharmaceutically acceptable carrier. 
     
     
         67 . A method of transfecting cells comprising contacting cells with a polynucleotide formulated with a particle comprising an activated polymer comprising one or more hydrolysed backbone esters.