US2023136645A1PendingUtilityA1

Compositions and methods for phosphoramidite and oligonucleotide synthesis

Assignee: WAVE LIFE SCIENCES LTDPriority: Mar 13, 2016Filed: Aug 5, 2022Published: May 4, 2023
Est. expiryMar 13, 2036(~9.7 yrs left)· nominal 20-yr term from priority
C07H 1/06C07H 19/16C07H 19/06C07F 9/65844C07H 19/173C07H 19/073C07H 23/00C07D 413/12C07D 417/14C07D 471/04C07D 405/14
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Claims

Abstract

The present disclosure, among other things, provides technologies for preparing and purifying phosphoramidites for oligonucleotide synthesis.

Claims

exact text as granted — not AI-modified
1 - 15 . (canceled) 
     
     
         16 . A composition comprising:
 a) a pre-treated purification medium; and   b) a phosphoramidite; or   
       a composition comprising:
 a) silica gel pre-treated with a hygroscopic solvent; and 
 b) a phosphoramidite: 
 wherein: 
 
       the phosphoramidite has the structure of formula I, 
       
         
           
           
               
               
           
         
       
       wherein:
 BA is R, or an optionally substituted group selected from a 3-30 membered cycloaliphatic ring, a 6-30 membered aryl ring, a 3-30 membered heterocyclyl having 1-10 heteroatoms independently selected from oxygen, nitrogen, sulfur, phosphorus and silicon, a 5-30 membered heteroaryl ring having 1-10 heteroatoms independently selected from oxygen, nitrogen, sulfur, phosphorus and silicon, a natural nucleobase moiety and a modified nucleobase moiety; 
 SU is a sugar moiety, a modified sugar moiety, -L-O— or 
 
       
         
           
           
               
               
           
         
          wherein SU is connected to the phosphorus atom in formula I through the oxygen atom; 
         L is a covalent bond, or a bivalent, optionally substituted, linear or branched group selected from C 1-30  aliphatic and C 1-30  heteroaliphatic group having 1-10 heteroatoms independently selected from oxygen, nitrogen, sulfur, phosphorus and silicon, wherein one or more methylene units are optionally and independently replaced by an optionally substituted C 1-6  alkylene, C 1-6  alkenylene, —C≡C—, —C(R′) 2 —, -Cy-, —O—, —S—, —S—S—, —N(R′)—, —C(O)—, —C(S)—, —C(NR′)—, —C(O)N(R′)—, —N(R′)C(O)N(R′)—, —N(R′)C(O)—, —N(R′)C(O)O—, —OC(O)N(R′)—, —S(O)—, —S(O) 2 —, —S(O) 2 N(R′)—, —N(R′)S(O) 2 —, —SC(O)—, —C(O)S—, —OC(O)—, or —C(O)O—; 
         R 5s  is R′ or —OR′; 
         R 2s  is —F, —CN, —N 3 , —NO, —NO 2 , -R′, —OR′, —SR′, —N(R′) 2 , -L-R′, —O-L-OR′, —O-L-SR′, or —O-L-N(R′) 2 , or R 2s  is L connecting C2 with C1, C2, C3, C4 or C5; 
         -Cy- is an optionally substituted bivalent ring selected from 3-30 membered carbocyclylene, 6-30 membered arylene, 5-30 membered heteroarylene having 1-10 heteroatoms independently selected from oxygen, nitrogen and sulfur, and 3-30 membered heterocyclylene having 1-10 heteroatoms independently selected from oxygen, nitrogen, sulfur, phosphorus and silicon; 
         R 1 , R 2 , and R 3  are taken together with their intervening atoms to form: 
       
       
         
           
           
               
               
           
         
         Ring A is an optionally substituted multivalent, monocyclic, bicyclic or polycyclic, saturated, partially unsaturated, or aryl 3-30 membered ring having, in addition to the intervening atoms, 0-10 heteroatoms independently selected from oxygen, nitrogen, sulfur, phosphorus and silicon; 
         each R s  is independently R′ or -L-R′; 
         t is 0-5; 
         each R′ is independently —R, —C(O)R, —CO 2 R, or —SO 2 R, or:
 two or more R′ are taken together with their intervening atoms to form an optionally substituted monocyclic, bicyclic or polycyclic, saturated, partially unsaturated, or aryl 3-30 membered ring having, in addition to the intervening atoms, 0-10 heteroatoms independently selected from oxygen, nitrogen, sulfur, phosphorus and silicon; and 
 each R is independently hydrogen, or an optionally substituted group selected from C 1-30  aliphatic, C 1-30  heteroaliphatic having 1-10 heteroatoms independently selected from oxygen, nitrogen, sulfur, phosphorus and silicon, a 6-30 membered aryl ring, a 5-30 membered heteroaryl ring having 1-10 heteroatoms independently selected from oxygen, nitrogen, sulfur, phosphorus and silicon, and a 3-30 membered heterocyclic ring having 1-10 heteroatoms independently selected from oxygen, nitrogen, sulfur, phosphorus and silicon. 
 
       
     
     
         17 - 19 . (canceled) 
     
     
         20 . The composition of  claim 16 , wherein the composition comprising:
 a) a pre-treated purification medium; and   b) a phosphoramidite.   
     
     
         21 . The composition of  claim 16 , wherein BA is a natural nucleobase moiety and a modified nucleobase moiety. 
     
     
         22 . The composition of claim  3 , wherein SU is 
       
         
           
           
               
               
           
         
       
     
     
         23 . The composition of  claim 22 , wherein R 5s  is —ODMTr. 
     
     
         24 . The composition of  claim 22 , wherein R 2s  is selected form —H, —F, —OMe, and —OCH 2 CH 2 OMe. 
     
     
         25 . The composition of  claim 16 , wherein 
       
         
           
           
               
               
           
         
         wherein one or more R 5s  is not hydrogen. 
       
     
     
         26 . The method of  claim 16 , wherein 
       
         
           
           
               
               
           
         
       
     
     
         27 . The composition of  claim 16 , wherein 
       
         
           
           
               
               
           
         
       
     
     
         28 . The composition of  claim 16 , wherein the phosphoramidite is selected from: 
       
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         or is selected from: 
       
       
         
           
           
               
               
           
         
          wherein X is 
       
       
         
           
           
               
               
           
         
          and: 
         R′ is H and R is H, 
         R′ is H, R is OMe, 
         R′ is H and R is OCH 2 CH 2 OMe, 
         R′ is H and R is F, 
         R′ is Me and R is H, 
         R′ is Me, R is OMe, 
         R′ is Me and R is OCH 2 CH 2 OMe, or 
         R′ is Me and R is F; 
       
       
         
           
           
               
               
           
         
          wherein X is 
       
       
         
           
           
               
               
           
         
          and 
         R′ is H and R is H, 
         R′ is H, R is OMe, 
         R′ is H and R is OCH 2 CH 2 OMe, 
         R′ is H and R is F, 
         R′ is Me and R is H, 
         R′ is Me, R is OMe, 
         R′ is Me and R is OCH 2 CH 2 OMe, or 
         R′ is Me and R is F; 
       
       
         
           
           
               
               
           
         
          wherein X is 
       
       
         
           
           
               
               
           
         
          and: 
         R′ is H and R is H, 
         R′ is H, R is OMe, 
         R′ is H and R is OCH 2 CH 2 OMe, 
         R′ is H and R is F, 
         R′ is Me and R is H, 
         R′ is Me, R is OMe, 
         R′ is Me and R is OCH 2 CH 2 OMe, or 
         R′ is Me and R is F; 
       
       
         
           
           
               
               
           
         
          wherein X is 
       
       
         
           
           
               
               
           
         
          and 
         R′ is H and R is H, 
         R′ is H, R is OMe, 
         R′ is H and R is OCH 2 CH 2 OMe, 
         R′ is H and R is F, 
         R′ is Me and R is H, 
         R′ is Me, R is OMe, 
         R′ is Me and R is OCH 2 CH 2 OMe, or 
         R′ is Me and R is F; 
       
       
         
           
           
               
               
           
         
          wherein X is 
       
       
         
           
           
               
               
           
         
          and: 
         R′ is Ac and R is H, 
         R′ is Ac, R is OMe, 
         R′ is Ac and R is OCH 2 CH 2 OMe, 
         R′ is Ac and R is F, 
         R′ is iBu and R is H, 
         R′ is iBu, R is OMe, 
         R′ is iBu and R is OCH 2 CH 2 OMe, or 
         R′ is iBu and R is F; 
       
       
         
           
           
               
               
           
         
          wherein X is 
       
       
         
           
           
               
               
           
         
          and: 
         R′ is Ac and R is H, 
         R′ is Ac, R is OMe, 
         R′ is Ac and R is OCH 2 CH 2 OMe, 
         R′ is Ac and R is F, 
         R′ is iBu and R is H, 
         R′ is iBu, R is OMe, 
         R′ is iBu and R is OCH 2 CH 2 OMe, or 
         R′ is iBu and R is F; 
       
       
         
           
           
               
               
           
         
          wherein X is 
       
       
         
           
           
               
               
           
         
          and: 
         R′ is Ac and R is H, 
         R′ is Ac, R is OMe, 
         R′ is Ac and R is OCH 2 CH 2 OMe, 
         R′ is Ac and R is F, 
         R′ is iBu and R is H, 
         R′ is iBu, R is OMe, 
         R′ is iBu and R is OCH 2 CH 2 OMe, or 
         R′ is iBu and R is F; 
       
       
         
           
           
               
               
           
         
          wherein X is 
       
       
         
           
           
               
               
           
         
          and: 
         R′ is Ac and R is H, 
         R′ is Ac, R is OMe, 
         R′ is Ac and R is OCH 2 CH 2 OMe, 
         R′ is Ac and R is F, 
         R′ is iBu and R is H, 
         R′ is iBu, R is OMe, 
         R′ is iBu and R is OCH 2 CH 2 OMe, or 
         R′ is iBu and R is F; 
       
       
         
           
           
               
               
           
         
          wherein X is 
       
       
         
           
           
               
               
           
         
          and: 
         R is H, 
         R is OMe, 
         R is OCH 2 CH 2 OMe, or 
         R is F; 
       
       
         
           
           
               
               
           
         
          wherein X is 
       
       
         
           
           
               
               
           
         
          and: 
         R is H, 
         R is OMe, 
         R is OCH 2 CH 2 OMe, or 
         R is F; 
       
       
         
           
           
               
               
           
         
          wherein X is 
       
       
         
           
           
               
               
           
         
          and: 
         R is H, 
         R is OMe, 
         R is OCH 2 CH 2 OMe, or 
         R is F; 
       
       
         
           
           
               
               
           
         
          wherein X is 
       
       
         
           
           
               
               
           
         
          and: 
         R is H, 
         R is OMe, 
         R is OCH 2 CH 2 OMe, or 
         R is F; 
       
       
         
           
           
               
               
           
         
          wherein X is 
       
       
         
           
           
               
               
           
         
          and: 
         R is H, 
         R is OMe, 
         R is OCH 2 CH 2 OMe, or 
         R is F; 
       
       
         
           
           
               
               
           
         
          wherein X is 
       
       
         
           
           
               
               
           
         
          and: 
         R is H, 
         R is OMe, 
         R is OCH 2 CH 2 OMe, or 
         R is F; 
       
       
         
           
           
               
               
           
         
          wherein X is 
       
       
         
           
           
               
               
           
         
          and: 
         R is H, 
         R is OMe, 
         R is OCH 2 CH 2 OMe, or 
         R is F; 
       
       
         
           
           
               
               
           
         
          wherein X is 
       
       
         
           
           
               
               
           
         
          and: 
         R is H, 
         R is OMe, 
         R is OCH 2 CH 2 OMe, or 
         R is F; 
       
       
         
           
           
               
               
           
         
          wherein X is 
       
       
         
           
           
               
               
           
         
          and: 
         R is H, 
         R is OMe, 
         R is OCH 2 CH 2 OMe, or 
         R is F; 
       
       
         
           
           
               
               
           
         
          wherein X is 
       
       
         
           
           
               
               
           
         
          and: 
         R is H, 
         R is OMe, 
         R is OCH 2 CH 2 OMe, or 
         R is F; 
       
       
         
           
           
               
               
           
         
          wherein X is 
       
       
         
           
           
               
               
           
         
          and: 
         R is H, 
         R is OMe, 
         R is OCH 2 CH 2 OMe, or 
         R is F; and 
       
       
         
           
           
               
               
           
         
          wherein X is 
       
       
         
           
           
               
               
           
         
          and: 
         R is H, 
         R is OMe, 
         R is OCH 2 CH 2 OMe, or 
         R is F. 
       
     
     
         29 . The composition of  claim 28 , wherein the phosphoramidite is selected from: 
       
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
       
     
     
         30 . The composition of  claim 28 , wherein the phosphoramidite is selected from: 
       
         
           
           
               
               
           
         
         wherein X is 
       
       
         
           
           
               
               
           
         
          and: 
         R′ is H and R is H, 
         R′ is H, R is OMe, 
         R′ is H and R is OCH 2 CH 2 OMe, 
         R′ is H and R is F, 
         R′ is Me and R is H, 
         R′ is Me, R is OMe, 
         R′ is Me and R is OCH 2 CH 2 OMe, or 
         R′ is Me and R is F; 
       
       
         
           
           
               
               
           
         
          wherein X is 
       
       
         
           
           
               
               
           
         
          and: 
         R′ is H and R is H, 
         R′ is H, R is OMe, 
         R′ is H and R is OCH 2 CH 2 OMe, 
         R′ is H and R is F, 
         R′ is Me and R is H, 
         R′ is Me, R is OMe, 
         R′ is Me and R is OCH 2 CH 2 OMe, or 
         R′ is Me and R is F; 
       
       
         
           
           
               
               
           
         
          wherein X is 
       
       
         
           
           
               
               
           
         
          and: 
         R′ is Ac and R is H, 
         R′ is Ac, R is OMe, 
         R′ is Ac and R is OCH 2 CH 2 OMe, 
         R′ is Ac and R is F, 
         R′ is iBu and R is H, 
         R′ is iBu, R is OMe, 
         R′ is iBu and R is OCH 2 CH 2 OMe, or 
         R′ is iBu and R is F; 
       
       
         
           
           
               
               
           
         
          wherein X is 
       
       
         
           
           
               
               
           
         
          and: 
         R′ is Ac and R is H, 
         R′ is Ac, R is OMe, 
         R′ is Ac and R is OCH 2 CH 2 OMe, 
         R′ is Ac and R is F, 
         R′ is iBu and R is H, 
         R′ is iBu, R is OMe, 
         R′ is iBu and R is OCH 2 CH 2 OMe, or 
         R′ is iBu and R is F; 
       
       
         
           
           
               
               
           
         
          wherein X is Ph 
       
       
         
           
           
               
               
           
         
          and: 
         R is H, 
         R is OMe, 
         R is OCH 2 CH 2 OMe, or 
         R is F; and 
       
       
         
           
           
               
               
           
         
          wherein X is 
       
       
         
           
           
               
               
           
         
          and: 
         R is H, 
         R is OMe, 
         R is OCH 2 CH 2 OMe, or 
         R is F. 
       
     
     
         31 . The composition of  claim 28 , wherein the phosphoramidite is selected from: 
       
         
           
           
               
               
           
         
         wherein X is 
       
       
         
           
           
               
               
           
         
          and: 
         R is H, 
         R is OMe, 
         R is OCH 2 CH 2 OMe, or 
         R is F; 
       
       
         
           
           
               
               
           
         
          wherein X is 
       
       
         
           
           
               
               
           
         
          and: 
         R is H, 
         R is OMe, 
         R is OCH 2 CH 2 OMe, or 
         R is F; 
       
       
         
           
           
               
               
           
         
          wherein X is 
       
       
         
           
           
               
               
           
         
          and: 
         R is H, 
         R is OMe, 
         R is OCH 2 CH 2 OMe, or 
         R is F; 
       
       
         
           
           
               
               
           
         
          wherein X is 
       
       
         
           
           
               
               
           
         
          and: 
         R is H, 
         R is OMe, 
         R is OCH 2 CH 2 OMe, or 
         R is F; 
       
       
         
           
           
               
               
           
         
          wherein X is 
       
       
         
           
           
               
               
           
         
          and: 
         R is H, 
         R is OMe, 
         R is OCH 2 CH 2 OMe, or 
         R is F; and 
       
       
         
           
           
               
               
           
         
          wherein X is 
       
       
         
           
           
               
               
           
         
          and: 
         R is H, 
         R is OMe, 
         R is OCH 2 CH 2 OMe, or 
         R is F. 
       
     
     
         32 . The composition of  claim 16 , wherein 
       
         
           
           
               
               
           
         
         wherein one R s  is hydrogen, and the other R s  is -L-R′, wherein R′ is —SO 2 R and L is a bivalent, optionally substituted C 1-30  aliphatic group.

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