US2018030448A1PendingUtilityA1

Pharmaceutical Composition Consisting Of RNA Having Alkali Metal As Counter Ion And Formulated With Dications

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Assignee: PASCOLO STEVEPriority: Dec 16, 2010Filed: Sep 25, 2017Published: Feb 1, 2018
Est. expiryDec 16, 2030(~4.4 yrs left)· nominal 20-yr term from priority
Inventors:Steve Pascolo
C12N 15/88A61K 47/02C12N 2320/32C12N 15/117C12N 2310/11C12N 15/11C12N 15/113A61K 9/0019C12N 2310/14C12N 15/111Y02A50/30
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Claims

Abstract

A biologically active RNA-alkali metal-dication formulation, a pharmaceutical composition containing the complexes, and methods of producing the same. The formulation is particularly useful to introduce RNA and an attached cargo into cells allowing its biological intracellular activities: e.g. immunostimulation (immunomodulation), RNA interference or gene expression.

Claims

exact text as granted — not AI-modified
1 . A composition comprising a complex of RNA and an alkali metal counter ion in a dication-containing solution, wherein the composition is prepared by the process of:
 (a) providing a RNA;   (b) next, preparing a RNA-alkali metal complex of the RNA and an alkali metal counter ion;   (c) then, adding a dication to the a RNA-alkali metal complex   
     
     
         2 . The composition of  claim 1  wherein the alkali metal counter ion is sodium. 
     
     
         3 . The composition of  claim 2  wherein the step of preparing the RNA-alkali metal complex of the RNA comprises ion exchange or precipitation. 
     
     
         4 . The composition of  claim 2  wherein the step of preparing the RNA-alkali metal complex comprises precipitation of the RNA-alkali metal complex in a mixture of sodium salt and an alcohol. 
     
     
         5 . The composition of  claim 2 , wherein the step of preparing the RNA-alkali metal complex comprises precipitation of the RNA-alkali metal complex in a mixture of sodium chloride or sodium acetate, and ethanol. 
     
     
         6 . The composition of  claim 1  wherein the dication is calcium. 
     
     
         7 . The composition of  claim 2  wherein the dication is calcium. 
     
     
         8 . The composition of  claim 1  wherein the RNA has a sequence containing more than 2 consecutive G residues or more than 4 consecutive U residues or GPurine (n) G wherein Purine is G or A and n from 1 to 4 or more. 
     
     
         9 . The composition of  claim 1  wherein the RNA has a sequence containing four or more consecutive G residues. 
     
     
         10 . The composition of  claim 6  wherein the RNA has a sequence containing four or more consecutive G residues. 
     
     
         11 . The composition of  claim 7  wherein the RNA has a sequence containing four or more consecutive G residues. 
     
     
         12 . The composition of  claim 8  wherein the RNA has a sequence containing four or more consecutive G residues. 
     
     
         13 . The composition of  claim 1  wherein the RNA has a sequence containing six consecutive G residues. 
     
     
         14 . The composition of  claim 1  wherein the RNA is linked to a chemical or biological cargo entity. 
     
     
         15 . A composition comprising an RNA-alkali metal complex of a RNA and an alkali metal counter ion in a dication-containing solution. 
     
     
         16 . The composition of  claim 15  wherein the RNA has a sequence containing four or more consecutive G residues. 
     
     
         17 . The composition of  claim 15  wherein the RNA has a sequence containing six consecutive G residues. 
     
     
         18 . The composition of  claim 15  wherein the alkali metal counter ion is sodium. 
     
     
         19 . The composition of  claim 15  wherein the dication is calcium. 
     
     
         20 . The composition of  claim 15  wherein the RNA is linked to a chemical or biological cargo entity. 
     
     
         21 . A method for the preparation of a complex of RNA and an alkali metal counter ion in a dication-containing solution, comprising:
 (a) providing a RNA;   (b) next, preparing a RNA-alkali metal complex of the RNA and an alkali metal counter ion;   (c) then, adding a dication to the a RNA-alkali metal complex   
     
     
         22 . The method  claim 21  wherein the step of preparing the RNA-alkali metal complex of the RNA comprises ion exchange or precipitation. 
     
     
         23 . The method of  claim 21  wherein the alkali metal counter ion is sodium. 
     
     
         24 . The method of  claim 23  wherein the step of preparing the RNA-alkali metal complex comprises precipitation of the RNA-alkali metal complex in a mixture of sodium salt and an alcohol. 
     
     
         25 . The method of  claim 23 , wherein the step of preparing the RNA-alkali metal complex comprises precipitation of the RNA-alkali metal complex in a mixture of sodium chloride or sodium acetate, and ethanol. 
     
     
         26 . The method of  claim 21  wherein the dication is calcium. 
     
     
         27 . The method of  claim 23  wherein the dication is calcium. 
     
     
         28 . The method of  claim 25 , wherein RNA-alkali metal complex is provided in an aqueous solution containing 0.2 to 20 mM calcium. 
     
     
         29 . The method of  claim 21 , wherein the RNA has a sequence containing more than 2 consecutive G residues or more than 4 consecutive U residues or GPurine (n) G wherein Purine is G or A and n from 1 to 4 or more. 
     
     
         30 . The method of  claim 29 , wherein the RNA has a sequence containing four or more consecutive G residues.

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