Process of purifying phosphoramidites
First Claim
Patent Images
1. A process of purifying a phosphitylated compound of formula I:
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wherein G1 is O or S;
G2 is a protecting group;
G3 is an amine;
Z is Z1 or Z2;
Z1 has the formula;
Z2 has the formula;
Y is O or S;
R′
2 is protected OH,;
H, HO, OR5, NR5R6, F, Cl, Br, alkyl, substituted alkyl, heterocyclyoalkyl, substituted heterocycloalkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, aralkyl, substituted aralkyl, heterocycloaralkyl, substituted heterocycloaralkyl;
or R′
2 and R′
4 together form a bridge;
each of R5 and R6 is independently alkyl, heterocycloalkyl, substituted heterocycloalkyl, aryl, substituted aryl, aralkyl, substituted aralkyl, hetaryl, substituted hetaryl, hetarylalkyl, substituted hetarylalkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, alkanoyl, substituted alkanoyl, trialkylsilyl or substituted trialkylsilyl;
orwhen R5 and R6 are on the same nitrogen, R5 and R6 may be taken together with the nitrogen to which they are bound to form a heterocyclyl ring or a substituted heterocyclyl ringR′
4 is H or together with R′
2 forms a bridge;
m is an integer of 0 or 1, Bx is a heterocycle and G4 is a hydroxy protecting group;
the process comprising;
(a) providing the compound of formula I in a polar phase comprising a polar organic solvent and at least one impurity;
(b) adding a basic compound and a first portion of water to the polar phase;
(c) contacting the polar phase with a first apolar organic phase;
(d) separating the first apolar organic phase from the polar phase;
(e) adding a second portion of water to the polar phase and contacting the polar phase with a second apolar organic phase, the polar phase and the second apolar phase being contacted for a time sufficient for the compound of formula I to partition into the second apolar organic phase; and
(f) separating the second apolar phase from the polar phase.
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Abstract
A process of purifying phosphoramidite precursors useful in inter alia synthesis of oligonucleotides comprises dissolving a crude phosphoramidite in a polar phase, adding a basic compound to the polar phase, adding a portion of water to the polar phase, contacting the polar phase with a first apolar phase to extract impurity into the apolar phase, separating the first apolar phase from the polar phase, adding a second aliquot of water to the polar phase, and contacting the polar phase with a second apolar phase, whereby the phosphoramidite partitions into the second apolar phase.
34 Citations
73 Claims
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1. A process of purifying a phosphitylated compound of formula I:
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wherein G1 is O or S; G2 is a protecting group; G3 is an amine; Z is Z1 or Z2; Z1 has the formula;
Z2 has the formula;
Y is O or S; R′
2 is protected OH,;
H, HO, OR5, NR5R6, F, Cl, Br, alkyl, substituted alkyl, heterocyclyoalkyl, substituted heterocycloalkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, aralkyl, substituted aralkyl, heterocycloaralkyl, substituted heterocycloaralkyl;
or R′
2 and R′
4 together form a bridge;each of R5 and R6 is independently alkyl, heterocycloalkyl, substituted heterocycloalkyl, aryl, substituted aryl, aralkyl, substituted aralkyl, hetaryl, substituted hetaryl, hetarylalkyl, substituted hetarylalkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, alkanoyl, substituted alkanoyl, trialkylsilyl or substituted trialkylsilyl;
orwhen R5 and R6 are on the same nitrogen, R5 and R6 may be taken together with the nitrogen to which they are bound to form a heterocyclyl ring or a substituted heterocyclyl ring R′
4 is H or together with R′
2 forms a bridge;m is an integer of 0 or 1, Bx is a heterocycle and G4 is a hydroxy protecting group;
the process comprising;(a) providing the compound of formula I in a polar phase comprising a polar organic solvent and at least one impurity; (b) adding a basic compound and a first portion of water to the polar phase; (c) contacting the polar phase with a first apolar organic phase; (d) separating the first apolar organic phase from the polar phase; (e) adding a second portion of water to the polar phase and contacting the polar phase with a second apolar organic phase, the polar phase and the second apolar phase being contacted for a time sufficient for the compound of formula I to partition into the second apolar organic phase; and (f) separating the second apolar phase from the polar phase. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25)
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26. A process of purifying a phosphitylated compound of formula I:
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wherein Bx is a heterocycle; Y is O or S; R′
2 is protected OH, H, OH, OR5, NR5R6, F, Cl, Br, alkyl, substituted alkyl, heterocyclyoalkyl, substituted heterocycloalkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, aralkyl, substituted aralkyl, heterocycloaralkyl, substituted heterocycloaralkyl;
or R′
2 and R′
4 together form a bridge;each of R5 and R6 is independently alkyl, heterocycloalkyl, substituted heterocycloalkyl, aryl, substituted aryl, aralkyl, substituted aralkyl, hetaryl, substituted hetaryl, hetarylalkyl, substituted hetarylalkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, alkanoyl, substituted alkanoyl, trialkylsilyl or substituted trialkylsilyl;
orwhen R5 and R6 are on the same nitrogen, R5 and R6 may be taken together with the nitrogen to which they are bound to form a heterocyclyl ring or a substituted heterocyclyl ring and R′
4 is H;
orR′
2 and R′
4 together form a bridge;G1 is O or S; G2 is a protecting group; G3 is an amine; and G4 is a protecting group; the process comprising; (a) providing the compound of formula I in a polar phase comprising a polar organic solvent and at least one impurity; (b) adding a basic compound and water to the polar phase; (c) contacting the polar phase with a first apolar organic phase; (d) separating the first apolar organic phase from the polar phase; (e) adding a second portion of water to the polar phase and contacting the polar phase with a second apolar organic phase, the polar phase and the second apolar phase being contacted for a time sufficient for the compound of formula I to partition into the second apolar organic phase; and (f) separating the second apolar phase from the polar phase. - View Dependent Claims (27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44)
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45. A process of purifying a phosphitylated compound of formula I:
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wherein G4 is a protecting group;
Bx is a nucleobase;R′ and
R″
each independently represent a substituted- or unsubstituted hydrocarbyl radical having 1 to 10 carbon atoms, or taken together R′ and
R″
, together with the nitrogen to which they are attached, form a nitrogen-containing heterocyclyl ring, which optionally contains one or two additional hetero atoms, said nitrogen-containing heterocyclyl ring being saturated or unsaturated;R′
2 is protected OH, H, OH, OR5, NR5R6, F, Cl, Br, alkyl, substituted alkyl, heterocyclyoalkyl, substituted heterocycloalkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, aralkyl, substituted aralkyl, heterocycloaralkyl, substituted heterocycloaralkyl;
orR′
2 and R′
4 together form a bridge;each of R5 and R6 is independently alkyl, heterocycloalkyl, substituted heterocycloalkyl, aryl, substituted aryl, aralkyl, substituted aralkyl, hetaryl, substituted hetaryl, hetarylalkyl, substituted hetarylalkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, alkanoyl, substituted alkanoyl, trialkylsilyl or substituted trialkylsilyl;
orwhen R5 and R6 are on the same nitrogen, R5 and R6 may be taken together with the nitrogen to which they are bound to form a heterocyclyl ring or a substituted heterocyclyl ring R′
4 is H or together with R′
2 forms a bridge; andR5 represents a protecting group; the process comprising; (a) providing the compound of formula I in a polar solution comprising a polar organic solvent; (b) adding a basic compound and a first portion of water to the polar solution; (c) contacting the polar phase with a first apolar organic phase; (d) removing the first apolar organic solvent from the polar phase; (e) adding a second portion of water to the polar phase and contacting the polar phase with a second apolar organic phase, said polar phase and said apolar organic phase remaining in contact for a period sufficient to extract the compound of formula I into the second apolar organic phase; and (f) separating the second apolar organic phase from the polar phase. - View Dependent Claims (46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68)
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69. A process of purifying a phosphoramidite by reverse precipitation at room temperature, said process comprising:
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(a) combining an intermediate purity phosphoramidite with a volume of an alkylated benzene solvent to form an organic solution; (b) gradually adding to the organic solution an apolar organic solvent until the phosphoramidite forms a gum phase that is discrete from the organic solution; and (c) separating the organic solution from the gum phase, said gum phase containing a purified phosphoramidite. - View Dependent Claims (70, 71, 72, 73)
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Specification