Human cytomegalovirus rna vaccines

Abstract

The disclosure describes HCMV ribonucleic acid (RNA) vaccines, as well as methods of using the vaccines and compositions comprising the vaccines.

1 - 96 . (canceled) 
     
     
         97 . A messenger ribonucleic acid (mRNA) vaccine comprising: an mRNA polynucleotide comprising an open reading frame encoding a human cytomegalovirus (hCMV) protein and a lipid nanoparticle comprising a compound of Formula (I): 
       
         
           
           
               
               
           
         
         or a salt or isomer thereof, wherein: 
         R 1  is selected from the group consisting of C 5-30  alkyl, C 5-20  alkenyl, —R*YR″, —YR″, and —R″M′R′; 
         R 2  and R 3  are independently selected from the group consisting of H, C 1-14  alkyl, C 2-14  alkenyl, —R*YR″, —YR″, and —R*OR″, or R 2  and R 3 , together with the atom to which they are attached, form a heterocycle or carbocycle; 
         R 4  is selected from the group consisting of a C 3-6  carbocycle, —(CH 2 ) n Q, —(CH 2 ) n CHQR, 
         CHQR, —CQ(R) 2 , and unsubstituted C 1-6  alkyl, where Q is selected from a carbocycle, heterocycle, —OR, —O(CH 2 ) n N(R) 2 , —C(O)OR, —OC(O)R, —CX 3 , —CX 2 H, —CXH 2 , —CN, —N(R) 2 , —C(O)N(R) 2 , —N(R)C(O)R, —N(R)S(O) 2 R, —N(R)C(O)N(R) 2 , —N(R)C(S)N(R) 2 , —N(R)R 8 , —O(CH 2 ) n OR, —N(R)C(═NR 9 )N(R) 2 , —N(R)C(═CHR 9 )N(R) 2 , —OC(O)N(R) 2 , —N(R)C(O)OR, —N(OR)C(O)R, —N(OR)S(O) 2 R, —N(OR)C(O)OR, —N(OR)C(O)N(R) 2 , —N(OR)C(S)N(R) 2 , —N(OR)C(═NR 9 )N(R) 2 , —N(OR)C(═CHR 9 )N(R) 2 , —C(═NR 9 )N(R) 2 , —C(═NR 9 )R, —C(O)N(R)OR, and —C(R)N(R) 2 C(O)OR, and each n is independently selected from 1, 2, 3, 4, and 5; 
         each R 5  is independently selected from the group consisting of C 1-3  alkyl, C 2-3  alkenyl, and H; 
         each R 6  is independently selected from the group consisting of C 1-3  alkyl, C 2-3  alkenyl, and H; 
         M and M′ are independently selected from —C(O)O—, —OC(O)—, —C(O)N(R′)—, 
         —N(R′)C(O)—, —C(O)—, —C(S)—, —C(S)S—, —SC(S)—, —CH(OH)—, —P(O)(OR′)O—, —S(O) 2 —, —S—S—, an aryl group, and a heteroaryl group; 
         R 7  is selected from the group consisting of C 1-3  alkyl, C 2-3  alkenyl, and H; 
         R 8  is selected from the group consisting of C 3-6  carbocycle and heterocycle; 
         R 9  is selected from the group consisting of H, CN, NO 2 , C 1-6  alkyl, —OR, —S(O) 2 R, —S(O) 2 N(R) 2 , C 2-6  alkenyl, C 3-6  carbocycle and heterocycle; 
         each R is independently selected from the group consisting of C 1-3  alkyl, C 2-3  alkenyl, and H; 
         each R′ is independently selected from the group consisting of C 1-18  alkyl, C 2-18  alkenyl, —R*YR″, —YR″, and H; 
         each R″ is independently selected from the group consisting of C 3-14  alkyl and C 3-14  alkenyl; 
         each R* is independently selected from the group consisting of C 1-12  alkyl and C 2-12  alkenyl; 
         each Y is independently a C 3-6  carbocycle; 
         each X is independently selected from the group consisting of F, Cl, Br, and I; and 
         m is selected from 5, 6, 7, 8, 9, 10, 11, 12, and 13. 
       
     
     
         98 . The mRNA vaccine of  claim 97 , wherein when R 4  is —(CH 2 ) n Q, —(CH 2 ) n CHQR, —CHQR, or —CQ(R) 2 , then (i) Q is not —N(R) 2  when n is 1, 2, 3, 4 or 5, or (ii) Q is not 5, 6, or 7-membered heterocycloalkyl when n is 1 or 2. 
     
     
         99 . The mRNA vaccine of  claim 97 , wherein:
 R 1  is selected from the group consisting of C 5-30  alkyl, C 5-20  alkenyl, —R*YR″, —YR″, and —R″M′R′;   R 2  and R 3  are independently selected from the group consisting of H, C 1-14  alkyl, C 2-14  alkenyl, —R*YR″, —YR″, and —R*OR″, or R 2  and R 3 , together with the atom to which they are attached, form a heterocycle or carbocycle;   R 4  is selected from the group consisting of a C 3-6  carbocycle, —(CH 2 ) n Q, —(CH 2 ) n CHQR, —CHQR, —CQ(R) 2 , and unsubstituted C 1-6  alkyl, where Q is selected from a C 3-6  carbocycle, a 5- to 14-membered heteroaryl having one or more heteroatoms selected from N, O, and S, —OR, —O(CH 2 ) n N(R) 2 , —C(O)OR, —OC(O)R, —CX 3 , —CX 2 H, —CXH 2 , —CN, —C(O)N(R) 2 , —N(R)C(O)R, —N(R)S(O) 2 R, —N(R)C(O)N(R) 2 , —N(R)C(S)N(R) 2 , —CRN(R) 2 C(O)OR, —N(R)R 8 , —O(CH 2 ) n OR, —N(R)C(═NR 9 )N(R) 2 , —N(R)C(═CHR 9 )N(R) 2 , —OC(O)N(R) 2 , —N(R)C(O)OR, —N(OR)C(O)R, —N(OR)S(O) 2 R, —N(OR)C(O)OR, —N(OR)C(O)N(R) 2 , —N(OR)C(S)N(R) 2 , —N(OR)C(═NR 9 )N(R) 2 , —N(OR)C(═CHR 9 )N(R) 2 , —C(═NR 9 )N(R) 2 , —C(═NR 9 )R, —C(O)N(R)OR, and a 5- to 14-membered heterocycloalkyl having one or more heteroatoms selected from N, O, and S which is substituted with one or more substituents selected from oxo (═O), OH, amino, mono- or di-alkylamino, and C 1-3  alkyl, and each n is independently selected from 1, 2, 3, 4, and 5;   each R 5  is independently selected from the group consisting of C 1-3  alkyl, C 2-3  alkenyl, and H;   each R 6  is independently selected from the group consisting of C 1-3  alkyl, C 2-3  alkenyl, and H;   M and M′ are independently selected from —C(O)O—, —OC(O)—, —C(O)N(R′)—, —N(R′)C(O)—, —C(O)—, —C(S)—, —C(S)S—, —SC(S)—, —CH(OH)—, —P(O)(OR′)O—, —S(O) 2 —, —S—S—, an aryl group, and a heteroaryl group;   R 7  is selected from the group consisting of C 1-3  alkyl, C 2-3  alkenyl, and H;   R 8  is selected from the group consisting of C 3-6  carbocycle and heterocycle;   R 9  is selected from the group consisting of H, CN, NO 2 , C 1-6  alkyl, —OR, —S(O) 2 R, —S(O) 2 N(R) 2 , C 2-6  alkenyl, C 3-6  carbocycle and heterocycle;   each R is independently selected from the group consisting of C 1-3  alkyl, C 2-3  alkenyl, and H;   each R′ is independently selected from the group consisting of C 1-18  alkyl, C 2-18  alkenyl, —R*YR″, —YR″, and H;   each R″ is independently selected from the group consisting of C 3-14  alkyl and C 3-14  alkenyl;   each R* is independently selected from the group consisting of C 1-12  alkyl and C 2-12  alkenyl;   each Y is independently a C 3-6  carbocycle;   each X is independently selected from the group consisting of F, Cl, Br, and I; and   m is selected from 5, 6, 7, 8, 9, 10, 11, 12, and 13,   or salts or isomers thereof.   
     
     
         100 . The mRNA vaccine of  claim 97 , wherein:
 R 1  is selected from the group consisting of C 5-30  alkyl, C 5-20  alkenyl, —R*YR″, —YR″, and —R″M′R′;   R 2  and R 3  are independently selected from the group consisting of H, C 1-14  alkyl, C 2-14  alkenyl, —R*YR″, —YR″, and —R*OR″, or R 2  and R 3 , together with the atom to which they are attached, form a heterocycle or carbocycle;   R 4  is selected from the group consisting of a C 3-6  carbocycle, —(CH 2 ) n Q, —(CH 2 ) n CHQR, —CHQR, —CQ(R) 2 , and unsubstituted C 1-6  alkyl, where Q is selected from a C 3-6  carbocycle, a 5- to 14-membered heterocycle having one or more heteroatoms selected from N, O, and S, —OR, —O(CH 2 ) n N(R) 2 , —C(O)OR, —OC(O)R, —CX 3 , —CX 2 H, —CXH 2 , —CN, —C(O)N(R) 2 , —N(R)C(O)R, —N(R)S(O) 2 R, —N(R)C(O)N(R) 2 , —N(R)C(S)N(R) 2 , —CRN(R) 2 C(O)OR, —N(R)R 8 , —O(C H 2 ) n OR, —N(R)C(═NR 9 )N(R) 2 , —N(R)C(═CHR 9 )N(R) 2 , —OC(O)N(R) 2 , —N(R)C(O)OR, —N(OR)C(O)R, —N(OR)S(O) 2 R, —N(OR)C(O)OR, —N(OR)C(O)N(R) 2 , —N(OR)C(S)N(R) 2 , —N(OR)C(═NR 9 )N(R) 2 , —N(OR)C(═CHR 9 )N(R) 2 , —C(═NR 9 )R, —C(O)N(R)OR, and —C(═NR 9 )N(R) 2 , and each n is independently selected from 1, 2, 3, 4, and 5; and when Q is a 5- to 14-membered heterocycle and (i) R 4  is —(CH 2 ) n Q in which n is 1 or 2, or (ii) R 4  is —(CH 2 ) n CHQR in which n is 1, or (iii) R 4  is —CHQR, and —CQ(R) 2 , then Q is either a 5- to 14-membered heteroaryl or 8- to 14-membered heterocycloalkyl;   each R 5  is independently selected from the group consisting of C 1-3  alkyl, C 2-3  alkenyl, and H;   each R 6  is independently selected from the group consisting of C 1-3  alkyl, C 2-3  alkenyl, and H;   M and M′ are independently selected from —C(O)O—, —OC(O)—, —C(O)N(R′)—, —N(R′)C(O)—, —C(O)—, —C(S)—, —C(S)S—, —SC(S)—, —CH(OH)—, —P(O)(OR′)O—, —S(O) 2 —, —S—S—, an aryl group, and a heteroaryl group;   R 7  is selected from the group consisting of C 1-3  alkyl, C 2-3  alkenyl, and H;   R 8  is selected from the group consisting of C 3-6  carbocycle and heterocycle;   R 9  is selected from the group consisting of H, CN, NO 2 , C 1-6  alkyl, —OR, —S(O) 2 R, —S(O) 2 N(R) 2 , C 2-6  alkenyl, C 3-6  carbocycle and heterocycle;   each R is independently selected from the group consisting of C 1-3  alkyl, C 2-3  alkenyl, and H;   each R′ is independently selected from the group consisting of C 1-18  alkyl, C 2-18  alkenyl, —R*YR″, —YR″, and H;   each R″ is independently selected from the group consisting of C 3-14  alkyl and C 3-14  alkenyl;   each R* is independently selected from the group consisting of C 1-12  alkyl and C 2-12  alkenyl;   each Y is independently a C 3-6  carbocycle;   each X is independently selected from the group consisting of F, Cl, Br, and I; and   m is selected from 5, 6, 7, 8, 9, 10, 11, 12, and 13,   or salts or isomers thereof.   
     
     
         101 . The mRNA vaccine of  claim 97 , wherein:
 R 1  is selected from the group consisting of C 5-30  alkyl, C 5-20  alkenyl, —R*YR″, —YR″, and —R″M′R′;   R 2  and R 3  are independently selected from the group consisting of H, C 1-14  alkyl, C 2-14  alkenyl, —R*YR″, —YR″, and —R*OR″, or R 2  and R 3 , together with the atom to which they are attached, form a heterocycle or carbocycle;   R 4  is selected from the group consisting of a C 3-6  carbocycle, —(CH 2 ) n Q, —(CH 2 ) n CHQR, —CHQR, —CQ(R) 2 , and unsubstituted C 1-6  alkyl, where Q is selected from a C 3-6  carbocycle, a 5- to 14-membered heteroaryl having one or more heteroatoms selected from N, O, and S, —OR, —O(CH 2 ) n N(R) 2 , —C(O)OR, —OC(O)R, —CX 3 , —CX 2 H, —CXH 2 , —CN, —C(O)N(R) 2 , —N(R)C(O)R, —N(R)S(O) 2 R, —N(R)C(O)N(R) 2 , —N(R)C(S)N(R) 2 , —CRN(R) 2 C(O)OR, —N(R)R 8 , —O(C H 2 ) n OR, —N(R)C(═NR 9 )N(R) 2 , —N(R)C(═CHR 9 )N(R) 2 , —OC(O)N(R) 2 , —N(R)C(O)OR, —N(OR)C(O)R, —N(OR)S(O) 2 R, —N(OR)C(O)OR, —N(OR)C(O)N(R) 2 , —N(OR)C(S)N(R) 2 , —N(OR)C(═NR 9 )N(R) 2 , —N(OR)C(═CHR 9 )N(R) 2 , —C(═NR 9 )R, —C(O)N(R)OR, and —C(═NR 9 )N(R) 2 , and each n is independently selected from 1, 2, 3, 4, and 5;   each R 5  is independently selected from the group consisting of C 1-3  alkyl, C 2-3  alkenyl, and H;   each R 6  is independently selected from the group consisting of C 1-3  alkyl, C 2-3  alkenyl, and H;   M and M′ are independently selected from —C(O)O—, —OC(O)—, —C(O)N(R′)—, —N(R′)C(O)—, —C(O)—, —C(S)—, —C(S)S—, —SC(S)—, —CH(OH)—, —P(O)(OR′)O—, —S(O) 2 —, —S—S—, an aryl group, and a heteroaryl group;   R 7  is selected from the group consisting of C 1-3  alkyl, C 2-3  alkenyl, and H;   R 8  is selected from the group consisting of C 3-6  carbocycle and heterocycle;   R 9  is selected from the group consisting of H, CN, NO 2 , C 1-6  alkyl, —OR, —S(O) 2 R, —S(O) 2 N(R) 2 , C 2-6  alkenyl, C 3-6  carbocycle and heterocycle;   each R is independently selected from the group consisting of C 1-3  alkyl, C 2-3  alkenyl, and H;   each R′ is independently selected from the group consisting of C 1-18  alkyl, C 2-18  alkenyl, —R*YR″, —YR″, and H;   each R″ is independently selected from the group consisting of C 3-14  alkyl and C 3-14  alkenyl;   each R* is independently selected from the group consisting of C 1-12  alkyl and C 2-12  alkenyl;   each Y is independently a C 3-6  carbocycle;   each X is independently selected from the group consisting of F, Cl, Br, and I; and   m is selected from 5, 6, 7, 8, 9, 10, 11, 12, and 13,   or salts or isomers thereof.   
     
     
         102 . The mRNA vaccine of  claim 97 , wherein:
 R 1  is selected from the group consisting of C 5-30  alkyl, C 5-20  alkenyl, —R*YR″, —YR″, and —R″M′R′;   R 2  and R 3  are independently selected from the group consisting of H, C 2-14  alkyl, C 2-14  alkenyl, —R*YR″, —YR″, and —R*OR″, or R 2  and R 3 , together with the atom to which they are attached, form a heterocycle or carbocycle;   R 4  is —(CH 2 ) n Q or —(CH 2 ) n CHQR, where Q is —N(R) 2 , and n is selected from 3, 4, and 5;   each R 5  is independently selected from the group consisting of C 1-3  alkyl, C 2-3  alkenyl, and H;   each R 6  is independently selected from the group consisting of C 1-3  alkyl, C 2-3  alkenyl, and H;   M and M′ are independently selected from —C(O)O—, —OC(O)—, —C(O)N(R′)—, —N(R′)C(O)—, —C(O)—, —C(S)—, —C(S)S—, —SC(S)—, —CH(OH)—, —P(O)(OR′)O—, —S(O) 2 —, —S—S—, an aryl group, and a heteroaryl group;   R 7  is selected from the group consisting of C 1-3  alkyl, C 2-3  alkenyl, and H;   each R is independently selected from the group consisting of C 1-3  alkyl, C 2-3  alkenyl, and H;   each R′ is independently selected from the group consisting of C 1-18  alkyl, C 2-18  alkenyl, —R*YR″, —YR″, and H;   each R″ is independently selected from the group consisting of C 3-14  alkyl and C 3-14  alkenyl;   each R* is independently selected from the group consisting of C 1-12  alkyl and C 1-12  alkenyl;   each Y is independently a C 3-6  carbocycle;   each X is independently selected from the group consisting of F, Cl, Br, and I; and   m is selected from 5, 6, 7, 8, 9, 10, 11, 12, and 13,   or salts or isomers thereof.   
     
     
         103 . The mRNA vaccine of  claim 97 , wherein:
 R 1  is selected from the group consisting of C 5-30  alkyl, C 5-20  alkenyl, —R*YR″, —YR″, and —R″M′R′;   R 2  and R 3  are independently selected from the group consisting of C 1-14  alkyl, C2-14 alkenyl, —R*YR″, —YR″, and —R*OR″, or R 2  and R 3 , together with the atom to which they are attached, form a heterocycle or carbocycle;   R 4  is selected from the group consisting of —(CH 2 ) n Q, —(CH 2 ) n CHQR, —CHQR, and —CQ(R) 2 , where Q is —N(R) 2 , and n is selected from 1, 2, 3, 4, and 5;   each R 5  is independently selected from the group consisting of C 1-3  alkyl, C 2-3  alkenyl, and H;   each R 6  is independently selected from the group consisting of C 1-3  alkyl, C 2-3  alkenyl, and H;   M and M′ are independently selected from —C(O)O—, —OC(O)—, —C(O)N(R′)—, —N(R′)C(O)—, —C(O)—, —C(S)—, —C(S)S—, —SC(S)—, —CH(OH)—, —P(O)(OR′)O—, —S(O) 2 —, —S—S—, an aryl group, and a heteroaryl group;   R 7  is selected from the group consisting of C 1-3  alkyl, C 2-3  alkenyl, and H;   each R is independently selected from the group consisting of C 1-3  alkyl, C 2-3  alkenyl, and H;   each R′ is independently selected from the group consisting of C 1-18  alkyl, C 2-18  alkenyl, —R*YR″, —YR″, and H;   each R″ is independently selected from the group consisting of C 3-14  alkyl and C 3-14  alkenyl;   each R* is independently selected from the group consisting of C 1-12  alkyl and C 1-12  alkenyl;   each Y is independently a C 3-6  carbocycle;   each X is independently selected from the group consisting of F, Cl, Br, and I; and   m is selected from 5, 6, 7, 8, 9, 10, 11, 12, and 13,   or salts or isomers thereof.   
     
     
         104 . The mRNA vaccine of  claim 97 , wherein the compound of Formula (I) comprises a compound of Formula (IA): 
       
         
           
           
               
               
           
         
       
       or a salt or isomer thereof, wherein 1 is selected from 1, 2, 3, 4, and 5; m is selected from 5, 6, 7, 8, and 9; M 1  is a bond or M′; R 4  is unsubstituted C 1-3  alkyl, or —(CH 2 ) n Q, in which Q is OH, —NHC(S)N(R) 2 , —NHC(O)N(R) 2 , —N(R)C(O)R, —N(R)S(O) 2 R, —N(R)R 8 , —NHC(═NR 9 )N(R) 2 , —NHC(═CHR 9 )N(R) 2 , —OC(O)N(R) 2 , —N(R)C(O)OR, heteroaryl or heterocycloalkyl; M and M′ are independently selected from —C(O)O—, —OC(O)—, —C(O)N(R′)—, —P(O)(OR′)O—, —S—S—, an aryl group, and a heteroaryl group; and R 2  and R 3  are independently selected from the group consisting of H, C 1-14  alkyl, and C 2-14  alkenyl. 
     
     
         105 . The mRNA vaccine of  claim 97 , wherein:
 R 1  is R″M′R′ or C 5-20  alkenyl;   R 2  and R 3  are each independently selected from C 1-14  alkyl and C 2-14  alkenyl;   R 4  is —(CH 2 ) n Q, wherein Q is OH and n is selected from 3, 4, and 5;   M and M′ are each independently —OC(O)— or —C(O)O—;   R 5 , R 6 , and R 7  are each H;   R′ is a linear C 1-12  alkyl, or C 1-12  alkyl substituted with C 6-9  alkyl;   R″ is C 3-14  alkyl; and   m is selected from 5, 6, 7, 8, 9, 10, 11, 12, and 13.   
     
     
         106 . The mRNA vaccine of  claim 105 , wherein:
 R 1  is R″M′R′;   R 2  and R 3  are each independently C 1-14  alkyl;   R 4  is —(CH 2 ) n Q, wherein Q is OH and n is 4;   M and M′ are each independently —OC(O)—;   R 5 , R 6 , and R 7  are each H;   R′ is C 1-12  alkyl substituted with C 6-9  alkyl;   R″ is C 3-14  alkyl; and   m is 6.   
     
     
         107 . The mRNA vaccine of  claim 105 , wherein:
 R 1  is C 5-20  alkenyl;   R 2  and R 3  are each independently C 1-14  alkyl;   R 4  is —(CH 2 ) n Q, wherein Q is OH and n is 3;   M is —C(O)O—;   R 5 , R 6 , and R 7  are each H; and   m is 6.   
     
     
         108 . The mRNA vaccine of  claim 97 , wherein the cationic lipid of Formula (I) is Compound 25: 
       
         
           
           
               
               
           
         
       
     
     
         109 . The mRNA vaccine of  claim 97 , wherein the lipid nanoparticle further comprises a PEG-modified lipid, a sterol, and a neutral lipid. 
     
     
         110 . The mRNA vaccine of  claim 109 , wherein the lipid nanoparticle comprises 20-60 mol % cationic lipid, 0.5-15 mol % PEG-modified lipid, 25-55 mol % sterol, and 5-25 mol % neutral lipid. 
     
     
         111 . The mRNA vaccine of  claim 110 , wherein the sterol is a cholesterol. 
     
     
         112 . The mRNA vaccine of  claim 97 , wherein the hCMV protein is selected from the group consisting of: gH, gL, UL128, UL130, UL131A, and gB. 
     
     
         113 . The mRNA vaccine of  claim 97 , comprising: an mRNA polynucleotide having an open reading frame encoding an hCMV gH protein; an mRNA polynucleotide having an open reading frame encoding an hCMV gL protein; an mRNA polynucleotide having an open reading frame encoding an hCMV UL128 protein; an mRNA polynucleotide having an open reading frame encoding an hCMV UL130 protein; an mRNA polynucleotide having an open reading frame encoding an hCMV UL131 protein; and an mRNA polynucleotide having an open reading frame encoding an hCMV gB protein. 
     
     
         114 . The mRNA vaccine of  claim 97 , wherein the mRNA polynucleotide comprises a chemical modification. 
     
     
         115 . The mRNA vaccine of  claim 114 , wherein the chemical modification is a N1-methylpseudouridine. 
     
     
         116 . A method of inducing an immune response in a subject, the method comprising administering to the subject the mRNA vaccine of  claim 97  in an amount effective to produce an antigen-specific immune response in the subject.