×

Process for the preparation of oligonucleotides

  • US 6,809,195 B1
  • Filed: 08/16/2000
  • Issued: 10/26/2004
  • Est. Priority Date: 08/16/2000
  • Status: Expired due to Term
First Claim
Patent Images

1. A method of preparing an oligomeric compound having at least one moiety of formula:

  • embedded imagewherein;

    X2 is O or S;

    X1 is Pg—

    O—

    , Pg—

    S—

    , C1-C10 straight or branched chain alkyl, CH3(CH2)nn

    O—

    , R2R3N—

    or a group remaining from coupling a chiral auxiliary;

    nn is from 0 to 10;

    Pg is CH3, —

    CH2CH2CN, —

    C(CH3)(CH3)—

    CCl3, —

    CH2

    CCl3, —

    CH2CH═

    CH2, CH2CH2SiCH3, 2-yl-ethyl phenylsulfonate, δ

    -cyanobutenyl, cyano p-xylyl, diphenylsilylethyl, 4-nitro-2-yl-ethylbenzene, 2-yl-ethyl-methyl sulfonate, methyl-N-trifluoroacetyl ethyl, acetoxy phenoxy ethyl, or a blocking group;

    R1 is, independently, hydrogen, a blocked hydroxyl group, or a sugar substituent group;

    R2 is, independently, hydrogen, a C1-C10 alkyl, a cycloalkyl, or aryl;

    R3 is, independently, hydrogen, a C1-C10 alkyl, a cycloalkyl, or aryl;

    R4 is, independently, N(L1)L2;

    or optionally, R2 and R3, together with the nitrogen atom to which they are attached form a cyclic moiety;

    each Bx is, independently, a heterocyclic base moiety; and

    comprising the steps of;

    (a) providing a 5′

    -O-protected compound of the formula;

    embedded imagewherein;

    T1 is a hydroxyl protecting group; and

    T2 is a covalent attachment to a support media, a nucleoside bound to a support media, a nucleotide, an oligonucleoside or an oligonucleotide;

    (b) treating said 5′

    -O-protected compound with a deprotecting reagent for a time and under conditions effective to form a 5′

    -deprotected compound;

    (c) coupling said 5′

    -O-deprotected compound with an activated phosphorus composition of the formula;

    embedded imagewherein;

    T3 is a hydroxyl protecting group, a nucleoside, nucleotide, an oligonucleoside or an oligonucleotide;

    each L1 and L2 is, independently, C1-6 straight or branched alkyl, or a C5-7 cyclic aliphatic ring system;

    or L1 and L2 are joined together to form a 4- to 13-membered heterocyclic ring system including the nitrogen atom to which L1 and L2 are attached; and

    R5 is X1;

    or R4 and R5 together with the phosphorus atom to which R4 and R5 are attached form a chiral auxiliary;

    for a time and under conditions effective to form an extended compound having the formula;

    embedded image(d) treating said extended compound with a mixture comprising an oxidizing reagent and a capping reagent in a single step and for a time and under conditions effective to form said oligomeric compound, and (e) treating said oligomeric compound with a reagent for a time and under conditions effective to remove said blocking groups thereby forming a deblocked oligomeric compound.

View all claims
  • 2 Assignments
Timeline View
Assignment View
    ×
    ×