Classifications

• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
  • C07H21/00—Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
  • C07H1/00—Processes for the preparation of sugar derivatives
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
  • C07H1/00—Processes for the preparation of sugar derivatives
  • C07H1/02—Phosphorylation
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
  • C07H19/00—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof
  • C07H19/02—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof sharing nitrogen
  • C07H19/04—Heterocyclic radicals containing only nitrogen atoms as ring hetero atom

Definitions

• Oligonucleotides may contain a variety of modifications. Certain modifications, such as phosphorothioate intemucleotidic linkages, may introduce new chiral centers into oligonucleotides.
• certain provided technologies utilize chiral auxiliaries that can be readily removed using bases without the utilization of HF, and/or basic conditions and/or elevated temperatures that may cause significant break of oligonucleotide chain and/or undesired transformation of certain intemucleotidic linkages (e.g., formation of natural phosphate linkage from phosphorothioate intemucleotidic linkages (or precursors thereof) and/or neutral intemucleotidic linkage (or precursors thereof)).
• Such technologies can provide new chemical compatibility, and can provide high stereoselectivity, cmde purity and yield as demonstrated herein.
• a useful compound is a compound of formula I, I-a, I-a-1, 1-a-2, 1-b, I-c, I-d, I-e, II, Il-a, Il-b, III, Ill-a, or III- b, or a salt thereof.
• such compounds may be utilized as chiral auxiliaries, e.g., for preparation of chirally controlled oligonucleotide compositions as demonstrated here.
• Oligonucleotide synthesis typically utilizes highly efficient chemical transformations in its steps.
• products of one or more steps often contain one or more reactive functional groups that can introduce significant impurities if uncapped, e.g., for coupling products, unreacted 5’-OH groups, and/or newly formed reactive groups (e.g., primary and/or secondary amino groups) when chiral auxiliaries are utilized for chirally controlled oligonucleotide synthesis.
• reactive functional groups are capped during oligonucleotide synthesis in order to reduce impurities from them.
• the present disclosure encompasses the recognition of a source of problem that capping steps as those typically used in traditional phosphoramidite-based oligonucleotide synthesis can lead to generation of a significant amount of byproducts, particularly for many chirally controlled (stereocontrolled, stereoselective) oligonucleotide synthesis processes.
• the present disclosure provides technologies that address the problems.
• the present disclosure provides methods comprising a post- modification capping step, e.g., after a modification step but before the next de-blocking and/or coupling step.
• a post-modification capping step is after a modification step that provides a chirally controlled intemucleotidic linkage but before the next de-blocking and/or coupling step.
• the present disclosure provides methods comprising capping steps of different chemistry strategies compared to a reference capping step in traditional oligonucleotide synthesis.
• the present disclosure provides methods comprising one or more capping steps, each of which selectively caps amino groups over hydroxyl groups (e.g., compared to a reference capping reagent system in traditional oligonucleotide synthesis).
• provided capping steps are selective for amidation over esterification.
• capping reagent systems for capping steps contain no or reduced levels (e.g., compared to a reference capping reagent system in traditional oligonucleotide synthesis) of strong nucleophiles and/or esterification catalysts (or reagents that can provide them when contacted with a composition to be capped), e.g., no or reduced levels of DMAP, NMI, etc..
• the present disclosure provides methods comprising capping steps that can cap both amino groups and hydroxyl groups efficiently, e.g., capping steps that are comparable or identical to a reference capping step in traditional oligonucleotide synthesis.
• the present disclosure provides methods that comprise capping steps of the same or different chemistry strategies to achieve oligonucleotide synthesis and can provide various advantages, e.g., improved crude purity, improved yield, etc., particularly for chirally controlled
• the present disclosure provides methods of a pre-modification capping step (after a coupling step but before the next modification step) and a post-modification capping step (after a modification step but before the next de blocking and/or coupling step).
• a pre-modification capping step and post- modification capping step are different.
• a pre-modification capping step and post- modification capping step have different chemistry strategies.
• a pre-modification capping step caps amino groups selectively over hydroxyl groups (e.g., compared to a reference capping reagent system in traditional oligonucleotide synthesis).
• a post-modification capping step can cap both amino and hydroxyl groups (e.g., compared to a reference capping reagent system in traditional oligonucleotide synthesis).
• a pre-modification capping step caps amino groups selectively over hydroxyl groups (e.g., compared to a reference capping reagent system in traditional oligonucleotide synthesis).
• a pre-modification capping step can cap both amino and hydroxyl groups (e.g., compared to a reference capping reagent system in traditional oligonucleotide synthesis).
• provided methods comprise two or more capping steps in an oligonucleotide synthesis cycle. In some embodiments, provided methods comprise two capping steps in an oligonucleotide synthesis cycle, wherein the two steps are separated by a modification step, e.g., oxidation, sulfurization, etc. In some embodiments, provided methods comprise a step in which a chiral modified intemucleotidic linkage comprising a chiral linkage phosphorus is formed with a
• the present disclosure provides a method comprising: providing a chiral nucleoside phosphoramidite which comprises a chiral atom that is not the phosphorus atom or a sugar carbon atom; and
• the present disclosure provides a method comprising:
• a chiral nucleoside phosphoramidite which comprises a chiral atom that is not the phosphorus atom and is not an atom of the nucleoside unit;
• the present disclosure provides a method comprising:
• oligonucleotide intermediate comprising a chiral linkage phosphorus atom, which is bonded to a chiral unit which does not comprise a nucleoside unit or a part thereof;
• the present disclosure provides a method comprising:
• oligonucleotide intermediate comprising a chiral linkage phosphorus atom, which is bonded to a chiral unit which does not comprise an atom of a nucleoside unit;
• the present disclosure provides a method, e.g., for preparing an oligonucleotide, comprising:
• a cycle comprises all optional steps.
• the present disclosure provides a method, e.g., for preparing a composition comprising a plurality of oligonucleotides comprising:
• a de-blocked composition comprising a plurality of de-blocked oligonucleotides (a de-blocked oligonucleotide composition) or nucleosides, which is de-blocked in that each independently comprises a free hydroxyl group, with a coupling reagent system comprising a partner compound which comprises a nucleoside unit; and
• the coupling step provides a coupling product composition
• a coupling product composition comprising a plurality of coupling product oligonucleotides, each of which independently comprises an intemucleotidic linkage connecting a hydroxyl group of a de-blocked oligonucleotide with a nucleoside unit of a partner compound;
• pre-modification capping step provides a pre -modification capping product composition comprising a plurality of pre-modification capping product oligonucleotides
• modification step provides a modification product composition comprising a plurality of modification product oligonucleotides
• post-modification capping step provides a post-modification capping product composition comprising a plurality of post-modification capping product oligonucleotides
• a de-blocking step comprising:
• the deblocking step provides a de-blocking product composition comprising a plurality of de-blocking product oligonucleotides, each of which independently comprises a free hydroxyl group;
• steps (1) through (5) optionally repeating steps (1) through (5) a number of times (e.g., 1-100, 1-90, 1-80, 1-70, 1- 60, 1-50, 1-40, 1-30, 1-25, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22,
• a number of times e.g., 1-100, 1-90, 1-80, 1-70, 1- 60, 1-50, 1-40, 1-30, 1-25, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22,
• provided methods comprise one or more pre-modification capping steps. In some embodiments, provided methods comprise one or more post-modification capping steps. In some embodiments, provided methods comprise one or more pre- and post-modification capping steps. In some embodiments, provided methods comprise one or more de-blocking steps.
• the present disclosure provides a method, e.g., for preparing an oligonucleotide, comprising one or more cycles, each of which independently comprises:
• the present disclosure provides a method, e.g., for preparing a composition comprising a plurality of oligonucleotides comprising one or more cycles, each cycle independently comprising:
• a de-blocked composition comprising a plurality of de-blocked oligonucleotides (a de-blocked oligonucleotide composition) or nucleosides, which is de-blocked in that each independently comprises a free hydroxyl group, with a coupling reagent system comprising a partner compound which comprises a nucleoside unit; and
• the coupling step provides a coupling product composition
• a coupling product composition comprising a plurality of coupling product oligonucleotides, each of which independently comprises an intemucleotidic linkage connecting a hydroxyl group of a de-blocked oligonucleotide with a nucleoside unit of a partner compound;
• pre-modification capping step provides a pre -modification capping product composition comprising a plurality of pre-modification capping product oligonucleotides
• a modification step comprising: contacting a coupling product composition with a modification reagent system comprising a modification reagent, and modifying one or more intemucleotidic linkages of one or more coupling product oligonucleotides; or
• modification step provides a modification product composition comprising a plurality of modification product oligonucleotides
• post-modification capping step provides a post-modification capping product composition comprising a plurality of post-modification capping product oligonucleotides
• a de-blocking step comprising:
• the deblocking step provides a de-blocking product composition comprising a plurality of de-blocking product oligonucleotides, each of which independently comprises a free hydroxyl group.
• a cycle comprises one or more pre-modification capping steps. In some embodiments, a cycle comprises one or more post-modification capping steps. In some embodiments, a cycle comprises one or more pre- and post-modification capping steps. In some embodiments, a cycle comprises one or more de-blocking steps. In some embodiments, a cycle comprises a coupling step, a pre -modification capping step, a modification step, a post-modification capping step, and a de-blocking step. In some embodiments, a cycle comprises a coupling step, a pre -modification capping step, a modification step, and a de-blocking step.
• a cycle comprises a coupling step, a modification step, a post-modification capping step and a de-blocking step.
• one or more cycles comprise a coupling step, a pre-modification capping step, a modification step, and a de-blocking step.
• one or more cycles comprise a coupling step, a modification step, a post-modification capping step and a de -blocking step.
• a cycle comprises one or more but not all optional steps.
• a cycle comprises a pre -modification step. In some embodiments, a cycle does not contain a pre-modification step. In some embodiments, a cycle comprises a post-modification step. In some embodiments, a cycle does not contain a post-modification step. In some embodiments, a cycle does not contain a de-blocking step, e.g., the last cycle when a desired length of an oligonucleotide is achieved.
• each step in a cycle is independently selected from a coupling step, a pre -modification capping step, a modification step, a post-modification capping, and a de-blocking step.
• a cycle may comprise two or more of the same steps (e.g., two coupling steps), each of which may utilize the same or different reagents, conditions, etc.
• steps e.g., two coupling steps
• repeat is performed before a next de-blocking step is performed.
• a coupling step is repeated immediately after another coupling step one or more times.
• a sequence of steps e.g., (l)-(2), (l)-(3), (l)-(2)-(3), (l)-(3)-(4), (l)-(2)-(3)-(4)) is repeated one or more times.
• “one or more” is one. In some embodiments,“one or more” is two or two or more. In some embodiments,“one or more” is 1-200, 1-
• “at least one” is one. In some embodiments,“at least one” is two or two or more. In some embodiments,“at least one” is 1-200, 1-150, 1-100, 1-90, 1-80, 1-70, 1-60, 1-50, 1-40, 1-30, 1-25, 1-24, 1-23, 1-22, 1-21, 1-20, 5-200, 5-150, 5-100, 5- 90, 5-80, 5-70, 5-60, 5-50, 5-40, 5-30, 5-25, 5-24, 5-23, 5-22, 5-21, 5-20, 10-200, 10-150, 10-100, 10-90, 10-80, 10-70, 10-60, 10-50, 10-40, 10-30, 10-25, 10-24, 10-23, 10-22, 10-21, 10-20, 15-200, 15-150, 15- 100, 15-90, 15-80, 15-70, 15-60, 15-50, 15-40, 15-30, 15-25, 15-24, 15-23, 15-22, 15-21, 15-20, 1, 2, 3, 4,
• one or more cycles comprise a coupling step, a pre-modification capping step, a modification step, a post-modification capping step, and a de-blocking step
• one or more cycles comprise a coupling step, a pre-modification capping step, a modification step, and a de- blocking step
• one or more cycles comprise a coupling step, a pre-modification capping step, a modification step, a post-modification capping step, and a de-blocking step
• one or more cycles comprise a coupling step, a modification step, a post-modification capping step and a de blocking step.
• a cycle consists of a coupling step, a pre -modification capping step, a modification step, a post-modification capping step, and a de-blocking step.
• a cycle consists of a coupling step, a pre-modification capping step, a modification step, and a de-blocking step.
• a cycle consists of a coupling step, a modification step, a post-modification capping step and a de -blocking step.
• a coupling step e.g., in a cycle
• a coupling step is immediately followed by a modification capping step.
• a coupling step e.g., in a cycle
• a pre-modification step e.g., in a cycle, is immediately followed by a modification step.
• a modification step comprises formation of a bond (e.g., single bond, double bond, etc.) between a linkage phosphorus and a sulfur or a nitrogen atom.
• a modification step is immediately followed by a post-modification capping step.
• a post- modification capping step is immediately followed by a de-blocking step.
• one or more washes may be included in steps described herein and may be performed before and/or after reactions of such steps, e.g., those steps in various methods and/or cycles described herein.
• suitable solvents in some embodiments, can be mixtures of chemicals
• oligonucleotides on solid support are typically extensive washed (e.g., to remove excess reagents, to remove undesired products, to switch solvents, to condition for a next reaction, etc.) before performance of another reaction of the same or an immediate following step.
• a step described herein e.g., a coupling step, a pre-modification capping step, a modification step, a post-modification capping step, or a de-blocking step, optionally comprises one or more washes.
• a reagent, solvent, and/or reagent system for a reaction is removed after the reaction is performed.
• removal is performed by filtration and/or washes when product oligonucleotides are on solid support, e.g., when using solid support for oligonucleotide synthesis.
• the present disclosure encompasses the recognition that traditional capping conditions when used as in traditional oligonucleotide synthesis may be a significant source of various problems under certain circumstances, and may contribute to formation of one or more by products (impurities) and significantly lower oligonucleotide crude purity and yield, particularly for stereoselective preparation of oligonucleotides comprising one or more chiral intemucleotidic linkages.
• the present disclosure provides technologies comprising capping strategies that can deliver unexpectedly high crude impurity and yield compared to an appropriate reference technology, for example, through designed capping strategies in combination with other steps in oligonucleotide synthesis.
• a reference technology uses a traditional capping condition as in traditional phosphoramidite-based oligonucleotide synthesis, which typically is or comprises an esterification condition that acrylates hydroxyl groups, e.g. , by using a mixture comprising an acylating agent (e.g., acetic anhydride), a base (e.g., 2,6-lutidine), and a catalyst (e.g., /V-methylimidazole, DMAP, etc.) to contact oligonucleotides to cap hydroxyl groups (e.g., unreacted 5’-OH groups).
• an acylating agent e.g., acetic anhydride
• a base e.g., 2,6-lutidine
• a catalyst e.g., /V-methylimidazole, DMAP, etc.
• acylating agent, base and catalyst for capping typically use a substantial amount of acylating agent, base and catalyst for capping, generally each independently about 5 %- 15% volume, and/or at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 30, 40, 50, 60, 70, 80, 90, or 100 equivalents relative to the first nucleoside incorporated into an oligonucleotide (before any cycle forming intemucleotidic linkage) or oligonucleotide loading capacity of a support (e.g., loading capacity of the support used for preparing an oligonucleotide, can be calculated by multiplying unit loading capacity of a support (e.g., umol/g) by amount of support (g)).
• the present disclosure provides technologies comprising one or more capping steps, e.g., pre-modification capping steps, post-modification capping steps, etc., each of which is independently comparable or identical to a reference capping step, e.g., of traditional oligonucleotide synthesis based on phosphoramidite chemistry.
• the present disclosure reduces by products that may be formed in such capping steps by strategically positioning their positions (or timing) in oligonucleotide synthesis methods and/or cycles.
• a typical condition selective or specific for amidation over esterification is an anhydride and a base without a catalyst (e.g., AC O and 2,6-lutidine), as a corresponding efficient esterification condition typically requires an anhydride, a base, and a catalyst (e.g., AC O, 2,6-lutidine, and NMI) as traditional capping conditions.
• the present disclosure provides technologies that comprise one or more synthetic cycles each independently comprising a coupling step, a modification step (e.g., oxidation, sulfiirization, etc.), and one or more capping steps, wherein each capping step after a coupling step and before a modification step comprising an amidation condition and no esterification condition.
• an amidation condition comprises no more than 0.01%, 0.02%, 0.05%, 0.1%, 0.2%, 0.5%, 1%, 2%, 3%, 4%, or 5% by volume of a catalyst for esterification under an appropriate corresponding condition (having the same acylating agent and base), and/or no more than about 0.01, 0.02, 0.05, 0.1, 0.2, 0.5, 1, 1.1, or 1.2 equivalents of a catalyst for esterification under an appropriate corresponding condition (having the same acylating agent and base), relative to an acylating agent and/or relative to
• an acylating agent is an anhydride. In some embodiments, an acylating agent is AC O. In some embodiments, a catalyst is NMI. In some embodiments, a catalyst is DMAP. In some embodiments, a catalyst is a nucleophilic nitrogen base.
• the present disclosure encompasses the recognition of a source of a problem in oligonucleotide synthesis, that an nucleophilic agent, particularly when used in a capping step that is after a coupling step and before a modification step in stereoselective oligonucleotide preparation, may contribute to generation of byproducts and lower overall preparation efficiency and/or crude purity through, e.g., degradation of oligonucleotides, lowering performance of another step, etc.
• each R N is independently R and the three R groups are taken together with the nitrogen atom to form an optionally substituted bicyclic or polycyclic ring as described in the present disclosure for R groups (and groups can be R), wherein the nitrogen of N(R n ) 3 (underlined) is a tertiary nitrogen, and there are no substitutions at any of the positions alpha to the nitrogen atom.
• a formed ring is saturated.
• a nucleophilic base is DABCO (l,4-diazabicyclo[2.2.2]octane).
• a formed ring contains one or more unsaturation.
• a nucleophilic nitrogen base is a compound of the structure of formula B-II:
• a formed ring is an optionally substituted C5-30 heteroaryl ring comprising 0-10 hetereoatoms in addition to the nitrogen atom.
• a formed ring is an optionally substituted 5-membered heteroaryl ring.
• a formed ring is a substituted 5-membered heteroaryl ring.
• a formed ring is a substituted imidazolyl ring.
• a nucleophilic base is substituted imidazole.
• a nucleophilic nitrogen base is NMI.
• a formed ring is an optionally substituted 6-membered heteroaryl ring.
• a formed ring is a substituted 6- membered heteroaryl ring.
• a formed ring is a substituted pyridinyl ring.
• a nucleophilic base is substituted pyridine.
• a nucleophilic nitrogen base is DMAP.
• nucleophilicity e.g., of basic nitrogen atoms in bases
• nucleophilicity is related to several factors, e.g., steric hindrance, electron density, etc. Technologies for assessing nucleophilicity are widely known in the art and can be utilized in accordance with the present disclosure. Additionally or alternatively, bases of various levels of nucleophilicity are well-known and can be assessed and/or utilized in accordance with the present disclosure.
• a base that can efficiently catalyze esterification reactions e.g., a base that can be used for efficient capping of unreacted 5’-OH together with anhydride and 2,6-lutidine in traditional oligonucleotide synthesis (e.g., DMAP, NMI, etc. ) is a strong nucleophilic base and should be avoided or used at reduced levels for capping steps that comprise greatly reduced levels of or no strong nucleophiles, e.g., any capping step after a coupling step and before a modification step.
• a strong nucleophilic base is a base that can effectively replace DMAP or NMI in esterification.
• a strong nucleophilic base is a base that can effectively replace DMAP or NMI in a capping step of traditional oligonucleotide synthesis (which typically uses phosphoramidite chemistry and does not use chiral auxiliaries and is considered non-stereoselective/non-stereocontrolled).
• provided methods comprise a capping step, which capping step comprises no more than 0.1, 0.2, 0.3, 0.4, 0.5, 0.5, 0.7, 0.8, 0.9, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 equivalents of a strong nucleophilic base relative to oligonucleotides or loading capacity of a support, or no strong nucleophilic bases.
• a capping step is immediately followed by a non-capping step.
• such a capping step is immediately after a non-capping step.
• such a capping step is immediately followed by a non-capping step, and is immediately after a non-capping step.
• a non-capping step is a coupling step. In some embodiments, a non-coupling step is a modification step. In some embodiments, a non-capping step immediately before such a capping step is a coupling modification step.
• each cycle independently forms an intemucleotidic linkage
• each cycle independently comprising a coupling step, one or more capping steps, and a modification step, which coupling step forms an intemucleotidic linkage, and which modification step modifies the intemucleotidic linkage formed in the coupling step;
• each capping step between the coupling step and the modification step comprises no strong nucleophile, or if it comprises one or more strong nucleophiles, level of each of the one or more strong nucleophiles is independently reduced compared to an appropriate reference capping condition.
• the present disclosure provides a method, e.g., for preparing an oligonucleotide, comprising one or more cycles, wherein:
• each cycle independently forms an intemucleotidic linkage
• a strong nucleophile is a strong nucleophile base as described in the present disclosure.
• a strong nucleophilic base is a compound of formula B-I.
• a strong nucleophilic base is a compound of formula B-I and can be used for efficient capping in traditional, phosphoramidite-based oligonucleotide synthesis.
• a strong nucleophilic base is a compound of formula B-II.
• a strong nucleophilic base is a compound of formula B-II and can be used for efficient capping in traditional, phosphoramidite-based oligonucleotide synthesis.
• a strong nucleophilic base is DMAP.
• the present disclosure provides a method, e.g., for preparing an oligonucleotide, comprising one or more cycles, wherein:
• each capping step between the coupling step and the modification step comprises no catalyst that promotes capping of 5’-OH as in an appropriate reference capping condition, or if it comprises one or more such catalysts, level of each of the one or more such catalysts is independently reduced compared to an appropriate reference capping condition.
• each cycle independently comprising a coupling step, one or more capping steps, and a modification step, which coupling step forms an intemucleotidic linkage, and which modification step modifies the intemucleotidic linkage formed in the coupling step;
• the present disclosure provides a method, e.g., for preparing an oligonucleotide, comprising one or more cycles, wherein:
• each cycle independently forms an intemucleotidic linkage
• each cycle independently comprising a coupling step, one or more capping steps, and a modification step, which coupling step forms an intemucleotidic linkage, and which modification step modifies the intemucleotidic linkage formed in the coupling step; wherein each capping step between the coupling step and the modification step (pre -modification capping step) comprises no catalyst for esterification, or if it comprises one or more catalysts for esterification, level of each of the one or more such catalysts is independently no more than 0.01, 0.02, 0.05, 0.1, 0.2, 0.5, or 1 equivalents relative to the first incorporated nucleoside of the oligonucleotide.
• the present disclosure provides a method, e.g., for preparing an oligonucleotide, comprising one or more cycles, wherein:
• each capping step between the coupling step and the modification step comprises a selective condition for amidation over esterification.
• each cycle independently comprising a coupling step, one or more capping steps, and a modification step, which coupling step forms an intemucleotidic linkage, and which modification step modifies the intemucleotidic linkage formed in the coupling step;
• each capping step between the coupling step and the modification step comprises a selective condition for amidation over esterification, and no condition identical to or comparable to an appropriate reference condition.
• selective conditions for amidation over esterification comprise reduced levels of or no catalysts for esterification, e.g., no DMAP, NMI, etc.
• a condition identical to or comparable to an appropriate reference condition can be used to replace capping conditions in traditional phosphoramidite-based oligonucleotide synthesis without significantly reducing (or with no more than 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, or 50% of reduction of) efficiency, cmde purity, and/or yield.
• provided methods comprise a second capping step after a modification step in one or more cycles.
• a second capping step comprises an esterification condition that is identical or comparable with a reference capping condition, e.g., in terms of capping unreacted 5’-OH in oligonucleotide synthesis.
• a strong nucleophile is DMAP orNMI.
• a strong nucleophile is DMAP.
• a strong nucleophile is NMI.
• an esterification catalyst is DMAP or NMI.
• an esterification catalyst is DMAP.
• an esterification catalyst is NMI.
• the present disclosure provides a method, e.g., for preparing an oligonucleotide, comprising one or more cycles each independently comprising:
• provided methods comprise repeating a number of steps or cycles, e.g., until a desired length (e.g., a desired length of an oligonucleotide or an intermediate) is achieved.
• a desired length e.g., a desired length of an oligonucleotide or an intermediate
• an order is, or comprises (l)-(2), ( l)-(3), (2)-(3), (3)-(4), (3)-(5), (4)-(5), (5)-( 1), (1)- (2)-(3), (l)-(3)-(4), (l)-(2)-(3)-(4) or any combination thereof (as appreciated by those skilled in the art, each of (l)-(5) independently represents a corresponding step of a method or a cycle, e.g., (1) - a coupling step; (2)- a pre -modification capping step or a first capping step; (3) - a modification step; (4) - a post- modification capping or a second capping step; (5)- a de-blocking step).
• an order is or comprises (l)-(2). In some embodiments, an order is or comprises (l)-(3). In some embodiments, an order is or comprises (2)-(3). In some embodiments, an order is or comprises (3)-(4). In some embodiments, an order is or comprises (3)-(5). In some embodiments, an order is or comprises (4)-(5).
• an order is or comprises (5)-(l)-(3)-(4). In some embodiments, an order is or comprises (5)-(l)-(2)-(3)-(4).
• a cycle or each cycle independently consists of steps (l)-(5). In some embodiments, a cycle or each cycle independently consists of steps (l)-(5) in the order of (l)-(2)-(3)-(4)- (5). In some embodiments, a cycle or each cycle independently consists of steps (l)-(5) in the order of (5)-(l)-(2)-(3)-(4). In some embodiments, a cycle or each cycle independently consists of steps (l)-(5) in the order of (l)-(2)-(4)-(3)-(5). In some embodiments, a cycle or each cycle independently consists of steps (l)-(5) in the order of (5)-(l)-(2)-(4)-(3).
• a provided method optionally or additionally comprises one or more cycles each independently comprising steps (1), (2), (3), and (5), optionally in that order. In some embodiments, a provided method optionally or additionally comprises one or more cycles each independently consisting of steps (1), (2), (3), and (5), optionally in that order. In some embodiments, such a cycle provides a natural phosphate linkage, e.g., optionally after cleavage, deprotection, etc. of an oligonucleotide. In some embodiments, such a cycle provides a non-chirally controlled phosphorothioate intemucleotidic linkage, e.g., optionally after cleavage, deprotection, etc.
• a provided method optionally or additionally comprises one or more cycles each independently comprising steps (1), (3), (4), and (5), optionally in that order. In some embodiments, a provided method optionally or additionally comprises one or more cycles each independently consisting of steps (1), (3), (4), and (5), optionally in that order. In some embodiments, such a cycle provides a natural phosphate linkage, e.g., optionally after cleavage, deprotection, etc. of an oligonucleotide. In some embodiments, such a cycle provides a non-chirally controlled phosphorothioate intemucleotidic linkage, e.g., optionally after cleavage, deprotection, etc.
• each such cycle independently provides a natural phosphate linkage, e.g., optionally after cleavage, deprotection, etc. of an oligonucleotide.
• each natural phosphate linkage in an oligonucleotide is independently formed by such a cycle optionally after cleavage, deprotection, etc. of an oligonucleotide.
• a first capping step is a pre -modification capping step as described in the present disclosure.
• a first capping step utilizes a capping reagent system that is a pre-modification capping reagent system.
• a second capping step is a post- modification capping step as described in the present disclosure.
• a second capping step utilizes a capping reagent system that is a post-modification capping reagent system.
• an appropriate reference capping condition is a capping condition of traditional oligonucleotide synthesis based on phosphoramidite chemistry. Example cycles for traditional phosphoramidite-based oligonucleotide synthesis are described below, wherein the illustrated
• a cycle of is a DPSE cycle depicted below (DPSE chiral auxiliary compounds: for different configurations of linkage phosphorus):
• Capping- 1 is a pre-modification or first capping step.
• Capping-2 is a post-modification or second capping step.
• cycle exit is after de-blocking before the next coupling.
• cycle exit is before de-blocking (e.g., to keep a 5’-blocking group such as DMTr on).
• ⁇ is nucleoside, nucleotide, or oligonucleotide on support (optionally linked to support via a linker).
• a reduced level of the present disclosure is no more than a percentage, e.g., 0.01%, 0.02%, 0.05%, 0.1%, 0.2%, 0.5%, 1%, 2%, 3%, 4%, 5%, etc., by volume of a capping reagent solution.
• a percentage is 0.01%.
• a percentage is 0.02%.
• a percentage is 0.05%.
• a percentage is 0.1%.
• a percentage is 0.2%.
• a percentage is 0.5%.
• a percentage is 1%.
• a percentage is 2%.
• a percentage is 3%.
• a percentage is 4%.
• a percentage is 5%.
• a reduced level is no more than about 0.01, 0.02, 0.05, 0.1, 0.2, 0.5, 1, 1.1, or 1.2 equivalents relative to a reference agent.
• a reference agent is an acylating agent.
• a reference agent is a support (by oligonucleotide loading capacity).
• a reduced level is no more than about 0.01, 0.02, 0.05, 0.1, 0.2, 0.5, 1, 1.1, or 1.2 equivalents relative to an acylating agent.
• a reduced level is no more than about 0.01, 0.02, 0.05, 0.1, 0.2, 0.5, or 1 equivalent relative to oligonucleotide loading capacity of a support. In some embodiments, a reduced level is no more than about 0.01 equivalent. In some embodiments, a reduced level is no more than about 0.02 equivalent. In some embodiments, a reduced level is no more than about 0.05 equivalent. In some embodiments, a reduced level is no more than about 0.1 equivalent. In some embodiments, a reduced level is no more than about 0.2 equivalent. In some embodiments, a reduced level is no more than about 0.5 equivalent. In some embodiments, a reduced level is no more than about 1 equivalent. In some embodiments, a reduced level is no more than about 1.1 equivalents. In some embodiments, a reduced level is no more than about 1.2 equivalents.
• provided technologies are particularly useful for preparing chirally controlled oligonucleotide compositions.
• provided technologies comprise formation of one or more chiral intemucleotidic linkages each independently comprising a chiral linkage phosphorus, wherein each of the chiral linkage phosphorus chiral center is independently formed with a stereoselectivity as described in the present disclosure, e.g., of at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%.
• phosphoramidites of traditional oligonucleotide synthesis are utilized for non-chirally controlled intemucleotidic linkages, and/or non-chiral intemucleotidic linkages.
• Suitable chiral auxiliaries and phosphoramidites for chirally controlled oligonucleotide synthesis that can be utilized in accordance with the present disclosure include those described in US 9598458, US 9744183, US 9605019, US 9394333, US 8859755, US 20130178612, US 8470987, US 8822671, US 20150211006, US 20170037399, US 20180216107, US 20180216108, US 20190008986, US 20180216107, US 20180216108, US 20190008986, WO 2017/015555, WO
• a phosphoramidite has the structure of formula IV, IV-a, IV-b, IV-c-1, IV-c-2, IV-d, IV-e, IVa, IVa-a, IVa-b, IVa-c-1, IVa-c-2, IVa-d, IVa-e, V, V-a, V-b, V-c-1, V-c-2, V-d, V-e, VI, Vl-a, Vl-b, VI-c-1, VI-c-2, Vl-d, or Vl-e, or a salt thereof, as described in the present disclosure.
• provided technologies comprising formation of an intemucleotidic linkage having the stmcture of formula VII, VII- a- 1, VII-a-2, Vll-b, VII-c, Vll-d, Vll-e, NL-n-1, NL- n-2, NL-n-3, NL-n-4, NL, NL-a-1, NL-a-2, NL-b-1, NL-b-2, NL-c-1, NL-c-2, NL-d-1, NL-d-2, or a salt form thereof, as described in the present disclosure.
• provided technologies provides oligonucleotides as intermediates and/or products.
• a provided oligonucleotide is of formula O-I or a salt thereof as described in the present disclosure.
• an oligonucleotide e.g., a final product, a product of a reaction, a product of a step, e.g., is one described in US 9598458, US 9744183, US 9605019, US 9394333, US 8859755, US 20130178612, US 8470987, US 8822671, US 20150211006, US 20170037399, US 20180216107, US 20180216108, US 20190008986, WO 2017/015555, WO
• provided intermediates and/or products are chirally controlled oligonucleotide compositions of a plurality of oligonucleotides of formula O-I or salts thereof.
• provided intermediates and/or products are chirally controlled oligonucleotide compositions of US 9598458, US 9744183, US 9605019, US 9394333, US 8859755, US 20130178612, US 8470987, US 8822671, US 20150211006, US
• the present disclosure provides a composition comprising:
• the present disclosure provides a composition comprising:
• a capping reagent system comprising a first compound having the structure of formula B-I or B-
• oligonucleotides each comprising at least one intemucleotidic linkage comprising a -C(0)-N(-)- moiety or a -P-S- moiety;
• the first compound is at a level of at least 0.1, 0.2, 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 50, or 100 equivalents relative to the plurality of oligonucleotides.
• the present disclosure provides a composition comprising:
• each intemucleotidic linkage of oligonucleotides of the plurality is independently an intemucleotidic linkage comprising a -C(0)-N(-)- moiety and a linkage phosphoms that is tetravalent;
• the first compound is at a level of at least 0.1, 0.2, 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 50, or 100 equivalents relative to the plurality of oligonucleotides.
• the plurality of oligonucleotides is a plurality of oligonucleotides of a modification product composition.
• oligonucleotide composition comprising a plurality of oligonucleotides:
• oligonucleotides of the plurality share the same base sequence
• oligonucleotides of the plurality share the same pattern of backbone linkages
• oligonucleotides of the plurality comprise at least one chirally controlled intemucleotidic linkage; wherein at least ((DS) Nc * 100)% of all oligonucleotides sharing the same base sequence in the composition are oligonucleotides of the plurality, wherein DS is at least 80%, 85%, 90%, 91%, 92%,
• Nc is the number of chirally controlled intemucleotidic linkage.
• oligonucleotides of the plurality share the same linkage phosphoms configuration (Rp or rip).
• a chirally controlled intemucleotidic linkage is referred to as a“stereodefined” intemucleotidic linkage.
• a first compound is NMI. In some embodiments, a first compound is DMAP.
• oligonucleotides of a plurality are attached to a support, e.g., a solid support used to prepare the oligonucleotides.
• a support e.g., a solid support used to prepare the oligonucleotides.
• oligonucleotides of a plurality equals loading capacity of the solid support they are attached to.
• oligonucleotides of a plurality are attached to a support via linkers.
• linkers are known in the art and may be utilized in accordance with the present disclosure; certain examples are described herein.
• a plurality of oligonucleotides share 1) a common base sequence, 2) a common pattern of backbone linkages, and 3) a common pattern of backbone phosphorus modifications, wherein the plurality of oligonucleotides share the same linkage phosphorus stereochemistry at one or more chiral intemucleotidic linkages (chirally controlled or stereodefined intemucleotidic linkages).
• about 1%-100% (e.g., 5%-100%, 10%-100%, 20%-100%, 30%-100%, 40%-100%, 50%-100%, 60%-100%, 70%-100%, 80-100%, 90-100%, 95-100%, 50%-90%, or about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%, or at least 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) of all intemucleotidic linkages in oligonucleotides of a plurality are chirally controlled intemucleotidic linkages.
• one or more intemucleotidic linkages are natural phosphate linkages. In some embodiments, all chiral intemucleotidic linkages which comprise chiral linkage phosphoms are chirally controlled intemucleotidic linkages.
• one or more chiral intemucleotidic linkages which comprise chiral linkage phosphoms are stereorandom chiral intemucleotidic linkages (not chirally controlled intemucleotidic linkages, typically prepared by non-chirally controlled methods, e.g., traditional oligonucleotide synthesis without utilization of chiral auxiliaries or chiral modification (e.g., sulfurization) reagents).
• oligonucleotides of a plurality share the same constitution.
• oligonucleotides of a plurality share the same stmcture (structurally identical).
• a plurality of oligonucleotides share the same stereochemistry at least one intemucleotidic linkage comprising a -C(0)-N(-)- moiety or a -P-S- moiety.
• about 0.1%-100% e.g., about 1%-100%, 5%-100%, 10%-100%, 20%-100%, 30%-100%, 40%-100%, 50%-100%, 60%-100%, 70%-100%, 80-100%, 90-100%, 95-100%, 50%-90%, or about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%, or at least 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) of all oligonucleotides in a chirally controlled oligonucleotide composition that share the common base sequence, or that share the common base sequence, the common pattern of backbone linkages, and the common pattern of backbone phosphorus modifications, or share the same constitution of oligonucle
• oligonucleotides of a plurality are each of the structure of formula O-I or a salt thereof.
• oligonucleotides of a plurality are oligonucleotides of US 9598458, US 9744183, US 9605019, US 9394333, US 8859755, US 20130178612, US 8470987, US 8822671, US 20150211006, US 20170037399, US 20180216107, US 20180216108, US 20190008986, WO 2017/015555, WO 2017/015575, WO 2017/062862, WO 2017/160741, WO 2017/192664, WO 2017/192679, WO 2017/210647, WO 2018/022473, WO 2018/067973, WO 2018/098264, WO
• oligonucleotides of each of which are independently incorporated herein by reference.
• oligonucleotides of a plurality share the same constitution.
• oligonucleotides of a plurality are identical.
• a level of the present disclosure is at least 0.1 equivalent. In some embodiments, a level is at least 0.2 equivalent. In some embodiments, a level is at least 0.5 equivalent.
• a level is at least 1 equivalent. In some embodiments, a level is at least 2 equivalents. In some embodiments, a level is at least 3 equivalents. In some embodiments, a level is at least 4 equivalents. In some embodiments, a level is at least 5 equivalents. In some embodiments, a level is at least 6 equivalents. In some embodiments, a level is at least 7 equivalents. In some embodiments, a level is at least 8 equivalents. In some embodiments, a level is at least 9 equivalents. In some embodiments, a level is at least 10 equivalents. In some embodiments, a level is at least 20 equivalents.
• a level is at least 50 equivalents. In some embodiments, a level is at least 100 equivalents.
• a -C(0)-N(-)- is part of a capped amino group in a chiral auxiliary moiety bonded to a linkage phosphorus, wherein the corresponding chiral auxiliary (replacing bonding to -C(O)- of -C(0)-N(-)- with -H, and replacing bonding to the linkage phosphorus with -H) is a compound of formula I, I-a, I-a-1, 1-a-2, 1-b, I-c, I-d, I-e, II, Il-a, Il-b, III, Ill-a, Ill-b, or a salt thereof.
• the present disclosure provides oligonucleotide compositions of high crude purity.
• the present disclosure provides a crude chirally controlled oligonucleotide composition comprising a plurality of oligonucleotides, wherein:
• oligonucleotides of the plurality share the same base sequence
• oligonucleotides of the plurality share the same pattern of backbone linkages
• oligonucleotides of the plurality comprise at least one chirally controlled intemucleotidic linkage, which intemucleotidic linkage is chirally controlled in that oligonucleotides of the plurality share the same stereochemical configuration at the chiral linkage phosphorus of the intemucleotidic linkage;
• oligonucleotides sharing the same base sequence in the crude composition are oligonucleotides of the plurality, wherein DS is at least 80%, 85%, 90%, 91%,
• Nc is the number of chirally controlled
• the present disclosure provides a cmde chirally controlled oligonucleotide composition comprising a plurality of oligonucleotides, wherein:
• oligonucleotides of the plurality share the same constitution
• oligonucleotides of the plurality comprise at least one chirally controlled intemucleotidic linkage, which intemucleotidic linkage is chirally controlled in that oligonucleotides of the plurality share the same stereochemical configuration at the chiral linkage phosphoms of the intemucleotidic linkage;
• oligonucleotides sharing the same base sequence in the cmde composition are oligonucleotides of the plurality, wherein DS is at least 80%, 85%, 90%, 91%,
• Nc is the number of chirally controlled
• a provided cmde chirally controlled oligonucleotide composition has a cmde purity of 30%-80%, 30%-90%, 30%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, or 75%, or more.
• a cmde chirally controlled oligonucleotide composition is cleaved from a support, and before any further purification.
• cmde chirally controlled oligonucleotide composition is cleaved from a support, after de-salting, and before any further purification.
• cmde chirally controlled oligonucleotide composition is before any chromatograph or gel purification.
• a cmde purity is % full-length product.
• a cmde purity is % full-length product as assessed by LC-UV monitored at UV 260 nm.
• DS is about 80%-100%, 85%-100%, 87%-100%, 89%-100%, 90- 100%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% or more. In some embodiments, DS is about 85% or more. In some embodiments, DS is about 86% or more. In some embodiments, DS is about 87% or more. In some embodiments, DS is about 88% or more. In some embodiments, DS is about 89% or more. In some embodiments, DS is about 90% or more. In some embodiments, DS is about 91% or more. In some embodiments, DS is about 92% or more.
• DS is about 93% or more. In some embodiments, DS is about 94% or more. In some embodiments, DS is about 95% or more. In some embodiments, DS is about 96% or more. In some embodiments, DS is about 97% or more. In some embodiments, DS is about 98% or more. In some embodiments, DS is about 99% or more.
• diastereoselectivity at, and/or diastereopurity of, chiral linkage phosphorus of a chiral intemucleotidic linkage in an oligonucleotide may be measured or represented through a model reaction, e.g.
• dimer under essentially the same or comparable conditions wherein the dimer has the same intemucleotidic linkage as the chiral intemucleotidic linkage, the 5’- nucleoside of the dimer is the same as the nucleoside to the 5’-end of the chiral intemucleotidic linkage, and the 3’-nucleoside of the dimer is the same as the nucleoside to the 3’-end of the chiral intemucleotidic linkage.
• diastereopurity of the underlined linkage in NNNNNNNG*SGNNNNNNN can be assessed from coupling two G moieties under the same or comparable conditions, e.g., monomers, chiral auxiliaries, solvents, activators, temperatures, etc.
• diastereopurity (and/or diastereoselectivity) of the linkage of a dimer (G*SG) is used as diastereopurity (and/or
• diastereopurity of a compound comprising multiple chiral elements is product of diastereomeric purity of all its chiral elements. In some embodiments, diastereopurity (i.e..
• diastereomeric purity of a provided oligonucleotide is product of diastereomeric purity of all its chiral linkage phosphoms in its chiral intemucleotidic linkages.
• Nc is the number of chirally controlled intemucleotidic linkage and is
• Nc is 1-50. In some embodiments, Nc is 1-40. In some embodiments, Nc is 1-30. In some embodiments, Nc is 1-25. In some embodiments, Nc is 1-24. In some embodiments, Nc is 1-23. In some embodiments, Nc is 1-22. In some embodiments, Nc is 1-21. In some embodiments, Nc is 1-20. In some embodiments, Nc is 1-19. In some embodiments, Nc is 1-18. In some embodiments, Nc is 1-17. In some embodiments, Nc is 1-16. In some embodiments, Nc is 1-15. In some embodiments, Nc is 1-14. In some embodiments, Nc is 1-13. In some embodiments, Nc is 1-12.
• Nc is 1-11. In some embodiments, Nc is 1-10. In some embodiments, Nc is 1-9. In some embodiments, Nc is 1-8. In some embodiments, Nc is 1-7. In some embodiments, Nc is 1-6. In some embodiments, Nc is 1-5. In some embodiments, Nc is 1. In some embodiments, Nc is 2. In some embodiments, Nc is 3. In some embodiments, Nc is 4. In some embodiments, Nc is 5. In some embodiments, Nc is 6. In some embodiments, Nc is 7. In some embodiments, Nc is 8. In some embodiments, Nc is 9. In some embodiments, Nc is 10. In some embodiments, Nc is 11. In some embodiments, Nc is 12.
• Nc is 13. In some embodiments, Nc is 14. In some embodiments, Nc is 15. In some embodiments, Nc is 16. In some embodiments, Nc is 17. In some embodiments, Nc is 18. In some embodiments, Nc is 19. In some embodiments, Nc is 20. In some embodiments, Nc is 21. In some embodiments, Nc is 22. In some embodiments, Nc is 23. In some embodiments, Nc is 24. In some embodiments, Nc is 25.
• provided technologies comprising one or more modification steps that independently comprise or are sulfurization (thiolation).
• provided intermediates and/or products comprise one or more phosphorothioate intemucleotidic linkages or precursors thereof (which can be converted into phosphorothioate intemucleotidic linkages upon deprotection/cleavage), optionally chirally controlled.
• provided intermediates and/or products comprise one or more non-negatively charged intemucleotidic linkages (e.g., neutral intemucleotidic linkages) or precursors thereof (which can be converted into phosphorothioate intemucleotidic linkages upon deprotection/cleavage), optionally chirally controlled.
• provided technologies comprising one or more modification steps that independently comprise or are oxidation.
• provided intermediates and/or products comprise one or more natural phosphate linkages or precursors thereof (which can be converted into natural phosphate linkages upon
• provided intermediates and/or products comprise one or more natural phosphate linkages and one or more phosphorothioate intemucleotidic linkages.
• Various supports can be utilized in accordance with the present disclosure, e.g., those described in US 9598458, US 9744183, US 9605019, US 9394333, US 8859755, US 20130178612, US 8470987, US 8822671, US 20150211006, US 20170037399, US 20180216107, US 20180216108, US 20190008986, WO 2017/015555, WO 2017/015575, WO 2017/062862, WO 2017/160741, WO
• 2019/032607 WO 2019/032612, WO 2019/032607, WO 2019/055951, WO 2019/075357, WO
• a support is a polymer. In some embodiments, a support is a solid support. In some embodiments, a solid support is a polymer, e.g., polystyrene. In some embodiments, a solid support is a Primer Support (e.g., Primer Support 5G, Primer Support 200, etc.). In some embodiments, a solid support is NittoPhase support (e.g.. Nitto Phase HU, NittoPhase UnyUinker, etc.). In some embodiments, a solid support is controlled-pore glass (CPG).
• CPG controlled-pore glass
• volume of a solid support e.g., certain polystyrene based solid support
• volume of a solid support changes during oligonucleotide synthesis, e.g., at different stages of synthesis and/or when contacted with different solvent systems and/or reagents.
• volume of a solid support e.g., many CPG support, changes less than 25%, 20%, 15%, 10%, or 5%, or remains substantially the same during oligonucleotide synthesis.
• the present disclosure encompasses the recognition than volume change of solid support during synthesis may cause deviations from planned reaction conditions, e.g., solvent system, reagent concentrations, contact time, etc., and may negatively impact synthesis efficiency, crude purity and/or yield.
• solid support that does not significantly change its volume or keep substantially the same volume during oligonucleotide synthesis may provide advantages, e.g., less deviation from planned reaction conditions, higher crude purity, higher yield, etc.
• a support can have a number of chemical modifications for nucleoside loading, and may have various unit loading capacities (e.g., umol/g).
• oligonucleotide synthesis using a support typically oligonucleotides are linked to a support through a linker.
• linkers can be utilized in accordance with the present disclosure, e.g., those described in US 9598458, US 9744183, US 9605019, US 9394333, US 8859755, US 20130178612, US 8470987, US 8822671, US 9403865, US 20150211006, US 20170037399, US 20180216107, US 20180216108, US 20190008986, WO 2017/015555, WO 2017/015575, WO 2017/062862, WO
• the present disclosure provides designed linkers. Useful support and/or functionalization there are described herein.
• Oligonucleotide synthesis typically comprises a deblocking step, which de-blocks a blocked hydroxyl group for a next step, e.g., a coupling step, which keeps intact capped hydroxyl groups which should not participate in a next step, e.g., a coupling step.
• a deblocking step which de-blocks a blocked hydroxyl group for a next step, e.g., a coupling step, which keeps intact capped hydroxyl groups which should not participate in a next step, e.g., a coupling step.
• de-blocking removes DMT group from DMT-protected hydroxyl (detritylation). In some embodiments, deblocking is performed by contacting oligonucleotides with an acid. In some
• an acid is trichloroacetic acid or dichloroacetic acid.
• a deblocking condition is 2% trichloroacetic acid (TCA) or 3% dichloroacetic acid (DCA), in an inert solvent (e.g., dichloromethane, toluene, etc.)
• a coupling step forms an intemucleotidic linkage, which adds a nucleoside unit to an existing oligonucleotide.
• an intemucleotidic linkage formed during a coupling step is a phosphite triester linkage.
• an intemucleotidic linkage can form with chirally control, e.g., as in chirally controlled oligonucleotide synthesis using diastereomerically pure
• phosphoramidite typically comprising a chiral auxiliary moiety.
• Conditions for coupling are widely reported and many can be utilized in accordance with the present disclosure, including those described in US 9598458, US 9744183, US 9605019, US 9394333, US 8859755, US 20130178612, US 8470987, US 8822671, US 20150211006, US 20170037399, US 20180216107, US 20180216108, US 20190008986, WO 2017/015555, WO 2017/015575, WO 2017/062862, WO 2017/160741, WO 2017/192664, WO 2017/192679, WO 2017/210647, WO 2018/022473, WO 2018/067973, WO 2018/098264, WO
• a coupling reagent system comprises a nucleoside phosphoramidite and an activator.
• Various phosphoramidites may be used in provided technologies, including those described in US 9598458, US 9744183, US 9605019, US 9394333, US 8859755, US 20130178612, US 8470987, US 8822671, US 20150211006, US 20170037399, US 20180216107, US 20180216108, US 20190008986, WO 2017/015555, WO 2017/015575, WO 2017/062862, WO 2017/160741, WO
• 2019/032607 WO 2019/032612, WO 2019/032607, WO 2019/055951 , WO 2019/075357, WO
• 2019/200185 or WO 2019/217784, and those having the structure of formula IV, IV-a, IV-b, IV-c-1, IV- c-2, IV-d, IV-e, IVa, IVa-a, IVa-b, IVa-c-1, IVa-c-2, IVa-d, IVa-e, V, V-a, V-b, V-c-1, V-c-2, V-d, V- e, VI, Vl-a, Vl-b, VI-c-1, VI-c-2, Vl-d, or Vl-e, or a salt thereof, as described in the present disclosure.
• Example activators include those described in US 9598458, US 9744183, US 9605019, US 9394333, US 8859755, US 20130178612, US 8470987, US 8822671, US 20150211006, US 20170037399, US 20180216107, US 20180216108, US 20190008986, WO 2017/015555, WO 2017/015575, WO
• an activator is CMPT. In some embodiments, an activator is CMIMT. In some embodiments, an activator is ETT. In some embodiments, conditions, e.g., concentrations of phosphoramidites, concentrations of activators, contact times, solvents, etc. can be optimized for each coupling to improve, e.g., crude purity, yield, etc.
• each capping step after a coupling step and before a modification step is performed as described in the present disclosure, e.g., with reduced levels of or no strong nucleophiles, with reduced levels of or no catalysts for esterification, and/or under conditions selective or specific for amidation over esterification.
• each capping step after a coupling step and before a modification step is performed to cap one or more amino groups, e.g., one or more amino groups formed after coupling in chiral auxiliaries moieties attached to linkage phosphorus atoms.
• a modification step is performed to modify a intemucleotidic linkage formed after coupling, which intemucleotidic linkage, in some embodiments, comprises a linkage phosphorus atom that is trivalent (e.g., as in a phosphite linkage).
• a modification step is or comprises sulfiirization.
• a nitrogen atom is in a salt form (quaternary) and is associated with a counterion (e.g., PF 6 ⁇ ).
• Example modifications and related technologies include those described in US 9598458, US 9744183, US
• 2019/032607 WO 2019/055951, WO 2019/075357, WO 2019/200185, or WO 2019/217784.
• reagents that tended to give inferior results in previously reported chirally controlled oligonucleotide synthesis can be utilized with technologies of the present disclosure to provide significantly improved, satisfactory results.
• the present disclosure provides technologies (e.g., chiral auxiliaries, compounds, methods, etc.) that are particularly effectively for chirally controlled preparation of oligonucleotides containing certain types of
• non-negatively charged intemucleotidic linkages e.g., nOOl
• an after-modification capping step is performed with a substantial amount of a strong nucleophile and/or an esterification catalyst (a strong nucleophile can be the same as an esterification catalyst) under an esterification condition which is comparable or identical to a capping condition in traditional oligonucleotide synthesis.
• an after-modification capping step caps free hydroxyl groups, e.g., those residue hydroxyl groups as a result of incomplete coupling which remain intact after a modification step. After this capping step, oligonucleotides can be de-blocked to expose hydroxyl groups at sites for further chain extension, and enter another synthetic cycle.
• oligonucleotides can be fully deprotected and cleaved from support for purification and/or further uses.
• Various cleavage and/or deprotection technologies can be utilized in accordance with the present disclosure, including those described in US 9598458, US 9744183, US 9605019, US 9394333, US 8859755, US 20130178612, US 8470987, US 8822671, US 20150211006, US 20170037399, US 20180216107, US 20180216108, US 20190008986, WO 2017/015555, WO 2017/015575, WO 2017/062862, WO 2017/160741, WO 2017/192664, WO 2017/192679, WO 2017/210647, WO 2018/022473, WO 2018/067973, WO 2018/098264, WO
• cleavage and/or deprotection comprise removal of chiral auxiliaries.
• cleavage and/or deprotection conditions can depend on the chemistry used during oligonucleotide synthesis, e.g., properties of linkers connecting
• removal of chiral auxiliaries comprises contacting oligonucleotides (e.g., a plurality of oligonucleotides) which comprise chiral auxiliaries with a base under an anhydrous condition, in some embodiments, preferably before the oligonucleotides are contacted with significant amount of water (e.g., a reagent system comprising a base, water, and optionally one or more organic solvents (e.g., one useful for deprotection of bases, cleavage of oligonucleotides from support, etc.)).
• water e.g., a reagent system comprising a base, water, and optionally one or more organic solvents (e.g., one useful for deprotection of bases, cleavage of oligonucleotides from support, etc.).
• water e.g., a reagent system comprising a base, water, and optionally one or more organic solvents (e.g., one
• deprotection/cleavage conditions comprising NH 3 .H 2 O
• deprotection/cleavage conditions comprising NH 3 .H 2 O
• removal of chiral auxiliaries using bases as described herein may simplify operation procedures and reduce manufacturing cost.
• sugars and nucleobases including non-natural, modified sugars and nucleobases, can be utilized in provided technologies in accordance with the present disclosure, e.g., those sugar and nucleobases described in US 9598458, US 9744183, US 9605019, US 9394333, US 8859755, US 20130178612, US 8470987, US 8822671, US 20150211006, US 20170037399, US 20180216107, US 20180216108, US 20190008986, WO 2017/015555, WO 2017/015575, WO
• a scale is 100 g or more. In some embodiments, a scale is 200 g or more. In some embodiments, a scale is 500 g or more. In some embodiments, a scale is at least 1000 g or more.
• a composition comprises a large scale of an oligonucleotide (e.g., a product of a reaction, a step, a method, etc. , or a final product).
• an oligonucleotide e.g., a product of a reaction, a step, a method, etc. , or a final product.
• the present disclosure addresses various challenges associated with large-scale preparation of oligonucleotide compositions, particularly chirally controlled oligonucleotide compositions.
• a de blocked composition being contacted in a coupling step is the de-blocked composition of the first de blocking step preceding the coupling step
• a coupling product composition being contacted in a pre- modification capping step is the coupling product composition of the first coupling step preceding the pre -modification capping step
• a coupling product composition being contacted in a modification step is the coupling product composition of the first coupling step preceding the modification step
• a pre- modification capping product composition being contacted in a modification step is the pre-modification capping product composition of the first pre-modification capping step preceding the modification step
• a modification product composition being contacted in a post-modification capping step is the modification product composition of the first modification step preceding the post-modification capping step
• a modification product composition being contacted in a de-blocking step is the modification product composition of the first modification step preceding the de-blocking step
• a post-modification capping product composition being contacted in a de-blocking step is the
• a method for preparing a composition comprising a plurality of oligomeric compounds comprises:
• a de-blocked composition comprising a plurality of de-blocked compounds, each independently comprising a de-blocked monomeric unit, which is de-blocked in that each de-blocked monomeric unit independently comprises a free connecting group, with a coupling reagent system which comprises a partner compound comprising a monomeric unit of the oligomeric compound;
• a de-blocking step comprising:
• a de-blocking reagent system to provide a de-blocked composition comprising a plurality of de-blocked products, each of which independently comprises a de-blocked monomeric unit comprising a free connecting group.
• a method optionally comprises repeating steps (1) to (5) a number of times, e.g., until a desired length is achieved.
• modification product composition comprising the chiral center and having the same constitution share the same stereochemical configuration at the chiral center, or a post-modification capping step, and a coupling reagent system comprising a chiral partner compound that comprises a monomeric unit of the oligomeric compound, wherein the chiral partner compound comprises a chiral atom that is not within the monomeric unit; or a coupling step which is immediately followed by a pre-modification capping step, which the pre-modification capping reagent system of the pre-modification capping step comprises no esterification catalyst or comprises no strong nucleophile.
• an oligomeric compound is an oligonucleotide.
• a composition comprising a plurality of oligomeric compounds is an oligonucleotide composition comprising a plurality of oligonucleotides.
• a coupling step is a coupling step as described in the present disclosure for oligonucleotide synthesis.
• a de-blocked compound is a de-blocked oligonucleotide, e.g., an oligonucleotide after de-blocking step in an oligonucleotide synthesis cycle.
• a de-blocked monomeric unit is a de-blocked 5’-end nucleoside unit.
• a free connecting group is a free 5’-hydroxyl group.
• a coupling reagent system is a coupling reagent system in oligonucleotide synthesis cycles.
• a partner compound is a phosphoramidite described herein for oligonucleotide synthesis.
• a coupling product is an oligonucleotide formed after coupling in oligonucleotide synthesis.
• a linkage linking the connecting group of a de -blocked monomeric unit and a monomeric unit of the partner compound is an intemucleotidic linkage formed during a coupling step.
• a pre- modification capping step is a capping step in oligonucleotide synthesis as described in the present disclosure.
• a pre -modification capping reagent system is a pre-capping reagent system in oligonucleotide synthesis as described in the present disclosure.
• a pre- modification capping product composition is a composition after a pre-modification capping step in oligonucleotide synthesis.
• a pre-modification capping product is a product formed after a pre-modification capping step in oligonucleotide synthesis.
• a modification step is a modification step as used in oligonucleotide synthesis.
• a modification step as demonstrated in oligonucleotide synthesis described in the present disclosure modifies an intemucleotidic linkage.
• a modification product composition is an oligonucleotide composition provided after a modification step in oligonucleotide synthesis.
• a modification product is an oligonucleotide provided after a modification step in oligonucleotide synthesis.
• a post-modification capping step is a capping step in oligonucleotide synthesis as described in the present disclosure.
• a post-modification capping reagent system is a post-capping reagent system in oligonucleotide synthesis as described in the present disclosure.
• a post-modification capping product composition is a composition after a post- modification capping step in oligonucleotide synthesis.
• a post-modification capping product is a product formed after a post-modification capping step in oligonucleotide synthesis.
• a de-blocking step is a de-blocking step as described in the present disclosure for oligonucleotide synthesis.
• a non-connecting group is an amino group.
• a chirally controlled chiral center is a chirally controlled linkage phosphorus center.
• a chiral partner compound comprising a chiral atom that is not within the monomeric unit is a phosphoramidite comprising a chiral center that is not in its nucleoside unit and is not the P.
• a reagent system comprises no esterification catalyst comprises no DMAP and no NMI.
• the term“a” or“an” may be understood to mean“at least one”;
• the term“or” may be understood to mean“and/or”;
• the terms“comprising”,“comprise”,“including” (whether used with“not limited to” or not), and “include” (whether used with“not limited to” or not) may be understood to encompass itemized components or steps whether presented by themselves or together with one or more additional components or steps;
• the term“another” may be understood to mean at least an additional/second one or more;
• the terms“about” and“approximately” may be understood to permit standard variation as would be understood by those of ordinary skill in the art; and (vi) where ranges are provided, endpoints are included.
• oligonucleotides and elements thereof e.g., base sequence, sugar modifications, intemucleotidic linkages, linkage phosphorus stereochemistry, etc.
• description of oligonucleotides and elements thereof is from 5’ to 3’.
• oligonucleotides described herein may be provided and/or utilized in a salt form, particularly a pharmaceutically acceptable salt form.
• oligonucleotides include various forms of the oligonucleotides.
• individual oligonucleotides within a composition may be considered to be of the same constitution and/or structure even though, within such composition (e.g., a liquid composition), particular such oligonucleotides might be in different forms (e.g., different pharmaceutically acceptable salt form(s) (and may be dissolved and the oligonucleotide chain may exist as an anion form when, e.g., in a liquid composition)) at a particular moment in time.
• individual intemucleotidic linkages along an oligonucleotide chain may be in an acid (H) form, or in one of a plurality of possible salt forms (e.g., a sodium salt, or a salt of a different cation, depending on which ions might be present in the preparation or composition), and will understand that, so long as their acid forms (e.g., replacing all cations, if any, with H + ) are of the same constitution and/or structure, such individual oligonucleotides may properly be considered to be of the same constitution and/or structure.
• Aliphatic Aliphatic.
• aliphatic means a straight-chain (i.e.. unbranched) or branched, substituted or unsubstituted hydrocarbon chain that is completely saturated or that contains one or more units of unsaturation, or a substituted or unsubstituted monocyclic, bicyclic, or polycyclic hydrocarbon ring that is completely saturated or that contains one or more units of unsaturation (but not aromatic), or combinations thereof.
• aliphatic groups contain 1-50 aliphatic carbon atoms. In some embodiments, aliphatic groups contain 1-20 aliphatic carbon atoms. In other embodiments, aliphatic groups contain 1-10 aliphatic carbon atoms.
• aliphatic groups contain 1-9 aliphatic carbon atoms. In other embodiments, aliphatic groups contain 1-8 aliphatic carbon atoms. In other embodiments, aliphatic groups contain 1-7 aliphatic carbon atoms. In other embodiments, aliphatic groups contain 1-6 aliphatic carbon atoms. In still other embodiments, aliphatic groups contain 1-5 aliphatic carbon atoms, and in yet other embodiments, aliphatic groups contain 1, 2, 3, or 4 aliphatic carbon atoms.
• Suitable aliphatic groups include, but are not limited to, linear or branched, substituted or unsubstituted alkyl, alkenyl, alkynyl groups and hybrids thereof such as (cycloalkyl)alkyl, (cycloalkenyl)alkyl or (cycloalkyl)alkenyl.
• Alkenyl refers to an aliphatic group, as defined herein, having one or more double bonds.
• Alkyl As used herein, the term“alkyl” is given its ordinary meaning in the art and may include saturated aliphatic groups, including straight-chain alkyl groups, branched-chain alkyl groups, cycloalkyl (alicyclic) groups, alkyl substituted cycloalkyl groups, and cycloalkyl substituted alkyl groups. In some embodiments, alkyl has 1-100 carbon atoms. In certain embodiments, a straight chain or branched chain alkyl has about 1-20 carbon atoms in its backbone (e.g., C1-C20 for straight chain, C2-C20 for branched chain), and alternatively, about 1-10.
• alkyl has 1-100 carbon atoms.
• a straight chain or branched chain alkyl has about 1-20 carbon atoms in its backbone (e.g., C1-C20 for straight chain, C2-C20 for branched chain), and alternatively, about 1-10.
• cycloalkyl rings have from about 3-10 carbon atoms in their ring structure where such rings are monocyclic, bicyclic, or polycyclic, and alternatively about 5, 6 or 7 carbons in the ring structure.
• an alkyl group may be a lower alkyl group, wherein a lower alkyl group comprises 1-4 carbon atoms (e.g., C1-C4 for straight chain lower alkyls).
• Alkynyl ⁇ As used herein, the term“alkynyl” refers to an aliphatic group, as defined herein, having one or more triple bonds.
• Analog includes any chemical moiety which differs structurally from a reference chemical moiety or class of moieties, but which is capable of performing at least one function of such a reference chemical moiety or class of moieties.
• a nucleotide analog differs structurally from a nucleotide but performs at least one function of a nucleotide
• a nucleobase analog differs structurally from a nucleobase but performs at least one function of a nucleobase; etc.
• Aryl ⁇ refers to monocyclic, bicyclic or polycyclic ring systems having a total of five to thirty ring members, wherein at least one ring in the system is aromatic.
• an aryl group is a monocyclic, bicyclic or polycyclic ring system having a total of five to fourteen ring members, wherein at least one ring in the system is aromatic, and wherein each ring in the system contains 3 to 7 ring members.
• an aryl group is a biaryl group.
• aryl may be used interchangeably with the term“aryl ring.”
• “aryl” refers to an aromatic ring system which includes, but not limited to, phenyl, biphenyl, naphthyl, binaphthyl, anthracyl and the like, which may bear one or more substituents.
• aryl also included within the scope of the term“aryl,” as it is used herein, is a group in which an aromatic ring is fused to one or more non-aromatic rings, such as indanyl, phthalimidyl, naphthimidyl, phenanthridinyl, or tetrahydronaphthyl, and the like, where a radical or point of attachment is on an aryl ring.
• Chiral control refers to control of the stereochemical designation of the chiral linkage phosphorus in a chiral intemucleotidic linkage within an oligonucleotide.
• a chiral intemucleotidic linkage is an intemucleotidic linkage whose linkage phosphoms is chiral.
• a control is achieved through a chiral element that is absent from the sugar and base moieties of an oligonucleotide, for example, in some embodiments, a control is achieved through use of one or more chiral auxiliaries during oligonucleotide preparation as described in the present disclosure, which chiral auxiliaries often are part of chiral phosphoramidites used during oligonucleotide preparation.
• oligonucleotide synthesis which does not use chiral auxiliaries cannot control stereochemistry at a chiral intemucleotidic linkage if such conventional oligonucleotide synthesis is used to form the chiral intemucleotidic linkage.
• the stereochemical designation of each chiral linkage phosphoms in each chiral intemucleotidic linkage within an oligonucleotide is controlled.
• Chirally controlled oligonucleotide composition refers to a composition that comprises a plurality of oligonucleotides (or nucleic acids) which share 1) a common base sequence, 2) a common pattern of backbone linkages, and 3) a common pattern of backbone phosphoms modifications, wherein the plurality of oligonucleotides (or nucleic acids) share the same linkage phosphoms stereochemistry at one or more chiral intemucleotidic linkages (chirally controlled or stereodefined intemucleotidic linkages, whose chiral linkage phosphoms is Rp or .S ' p in the composition (“stereodefined”), not a random Rp and .S ' p mixture as non-chirally controlled intemucleotidic linkages).
• Level of the plurality of oligonucleotides (or nucleic acids) in a chirally controlled oligonucleotide composition is pre-determined/controlled (e.g., through chirally controlled oligonucleotide preparation to stereoselective ly form one or more chiral intemucleotidic linkages).
• about 1%-100% (e.g., about 5%-100%, 10%-100%, 20%-100%, 30%-100%, 40%- 100%, 50%-100%, 60%-100%, 70%-100%, 80-100%, 90-100%, 95-100%, 50%-90%, or about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, or at least 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) of all oligonucleotides in a chirally controlled oligonucleotide composition are oligonucleotides of the plurality.
• about 1%-100% (e.g., about 5%-100%, 10%-100%, 20%-100%, 30%-100%, 40%-100%, 50%-100%, 60%-100%, 70%-100%, 80- 100%, 90-100%, 95-100%, 50%-90%, or about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, or at least 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) of all oligonucleotides in a chirally controlled oligonucleotide composition that share the common base sequence, the common pattern of backbone linkages, and the common pattern of backbone phosphoms modifications are oligonucleotides of the plurality.
• a level is about 1%-100%, (e.g., about 5%-100%, 10%-100%, 20%-100%, 30%-100%, 40%-100%, 50%-100%, 60%-100%, 70%- 100%, 80-100%, 90-100%, 95-100%, 50%-90%, or about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%,
• oligonucleotides in a composition or of all oligonucleotides in a composition that share a common base sequence (e.g., of a plurality of oligonucleotide or an oligonucleotide type), or of all oligonucleotides in a composition that share a common base sequence, a common pattern of backbone linkages, and a common pattern of backbone phosphorus modifications, or of all oligonucleotides in a composition that share a common base sequence, a common patter of base modifications, a common pattern of sugar modifications, a common pattern of in
• the plurality of oligonucleotides share the same stereochemistry at about 1-50 (e.g., about 1-10, 1-20, 5-10, 5-20, 10-15, 10-20, 10-25, 10-30, or about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20, or at least 1, 2, 3, 4, 5, 6, 7, 8, 9,
• the plurality of oligonucleotides share the same stereochemistry at about 1%-100% (e.g., about 5%-100%, 10%-100%, 20%-100%, 30%-100%, 40%-100%, 50%-100%, 60%-100%, 70%-100%, 80-100%, 90-
• oligonucleotides (or nucleic acids) of a plurality are of the same constitution.
• level of the oligonucleotides (or nucleic acids) of the plurality is about 1%-100%, (e.g., about 5%-100%, 10%-100%, 20%-100%, 30%-100%, 40%-100%, 50%-100%, 60%-100%, 70%-100%, 80-100%, 90-100%, 95-100%, 50%-90%, or about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, or at least 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) of all oligonucleotides (or nucleic acids) in a composition that share the same constitution as the oligonucleotides (or nucleic acids) of the plurality.
• each chiral intemucleotidic linkage is a chiral controlled intemucleotidic linkage, and the composition is a completely chirally controlled oligonucleotide composition.
• oligonucleotides (or nucleic acids) of a plurality are structurally identical.
• a chirally controlled intemucleotidic linkage has a diastereopurity of at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 99.5%, typically at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 99.5%.
• a chirally controlled intemucleotidic linkage has a diastereopurity of at least 95%.
• a chirally controlled intemucleotidic linkage has a diastereopurity of at least 96%.
• a chirally controlled intemucleotidic linkage has a diastereopurity of at least 97%. In some embodiments, a chirally controlled intemucleotidic linkage has a diastereopurity of at least 98%. In some embodiments, a chirally controlled intemucleotidic linkage has a diastereopurity of at least 99%.
• a percentage of a level is or is at least (DS) nc , wherein DS is a diastereopurity as described in the present disclosure (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 99.5% or more) and nc is the number of chirally controlled intemucleotidic linkages as described in the present disclosure (e.g., 1-50, 1-40, 1-30, 1-25, 1-20, 5-50, 5-40, 5-30, 5-25, 5-20, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25 or more).
• level of a plurality of oligonucleotides in a composition is represented as the product of the diastereopurity of each chirally controlled intemucleotidic linkage in the oligonucleotides.
• diastereopurity of an intemucleotidic linkage connecting two nucleosides in an oligonucleotide (or nucleic acid) is represented by the diastereopurity of an intemucleotidic linkage of a dimer connecting the same two nucleosides, wherein the dimer is prepared using comparable conditions, in some instances, identical synthetic cycle conditions (e.g., for the linkage between Nx and Ny in an oligonucleotide ....NxNy . , the dimer is
• not all chiral intemucleotidic linkages are chiral controlled intemucleotidic linkages, and the composition is a partially chirally controlled oligonucleotide composition.
• a non-chirally controlled intemucleotidic linkage has a diastereopurity of less than about 80%, 75%, 70%, 65%, 60%, 55%, or of about 50%, as typically observed in stereorandom oligonucleotide compositions (e.g. , as appreciated by those skilled in the art, from traditional oligonucleotide synthesis, e.g., the phosphoramidite method).
• oligonucleotides (or nucleic acids) of a plurality are of the same type.
• a chirally controlled oligonucleotide composition comprises non-random or controlled levels of individual oligonucleotide or nucleic acids types. For instance, in some embodiments a chirally controlled oligonucleotide composition comprises one and no more than one oligonucleotide type. In some embodiments, a chirally controlled oligonucleotide composition comprises more than one oligonucleotide type. In some embodiments, a chirally controlled oligonucleotide composition comprises multiple oligonucleotide types.
• a chirally controlled oligonucleotide composition is a composition of oligonucleotides of an oligonucleotide type, which composition comprises a non-random or controlled level of a plurality of oligonucleotides of the oligonucleotide type.
• Comparable is used herein to describe two (or more) sets of conditions or circumstances that are sufficiently similar to one another to permit comparison of results obtained or phenomena observed.
• comparable sets of conditions or circumstances are characterized by a plurality of substantially identical features and one or a small number of varied features.
• sets of conditions are comparable to one another when characterized by a sufficient number and type of substantially identical features to warrant a reasonable conclusion that differences in results obtained or phenomena observed under the different sets of conditions or circumstances are caused by or indicative of the variation in those features that are varied.
• Cycloaliphatic The term “cycloaliphatic,” “carbocycle,” “carbocyclyl,” “carbocyclic radical,” and“carbocyclic ring,” are used interchangeably, and as used herein, refer to saturated or partially unsaturated, but non-aromatic, cyclic aliphatic monocyclic, bicyclic, or polycyclic ring systems, as described herein, having, unless otherwise specified, from 3 to 30 ring members.
• Cycloaliphatic groups include, without limitation, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cycloheptyl, cycloheptenyl, cyclooctyl, cyclooctenyl, norbomyl, adamantyl, and cyclooctadienyl.
• a cycloaliphatic group has 3-6 carbons.
• a cycloaliphatic group is saturated and is cycloalkyl.
• cycloaliphatic may also include aliphatic rings that are fused to one or more aromatic or nonaromatic rings, such as decahydronaphthyl or tetrahydronaphthyl.
• a cycloaliphatic group is bicyclic.
• a cycloaliphatic group is tricyclic.
• a cycloaliphatic group is polycyclic.
• “cycloaliphatic” refers to C3-C6 monocyclic hydrocarbon, or CVCm bicyclic or polycyclic hydrocarbon, that is completely saturated or that contains one or more units of unsaturation, but which is not aromatic, that has a single point of attachment to the rest of the molecule, or a C 9 -C 16 polycyclic hydrocarbon that is completely saturated or that contains one or more units of unsaturation, but which is not aromatic, that has a single point of attachment to the rest of the molecule.
• Halogen means F, Cl, Br, or I.
• heteroaliphatic is given its ordinary meaning in the art and refers to aliphatic groups as described herein in which one or more carbon atoms are independently replaced with one or more heteroatoms (e.g ., oxygen, nitrogen, sulfur, silicon, phosphorus, and the like). In some embodiments, one or more units selected from C, CH, C3 ⁇ 4, and C3 ⁇ 4 are independently replaced by one or more heteroatoms (including oxidized and/or substituted forms thereof). In some embodiments, a heteroaliphatic group is heteroalkyl. In some embodiments, a heteroaliphatic group is heteroalkenyl.
• heteroalkyl refers to alkyl groups as described herein in which one or more carbon atoms are independently replaced with one or more heteroatoms (e.g., oxygen, nitrogen, sulfur, silicon, phosphorus, and the like).
• heteroalkyl groups include, but are not limited to, alkoxy, polyethylene glycol)-, alkyl- substituted amino, tetrahydrofuranyl, piperidinyl, morpholinyl, etc.
• Heteroaryl refers to monocyclic, bicyclic or polycyclic ring systems having a total of five to thirty ring members, wherein at least one ring in the system is aromatic and at least one aromatic ring atom is a heteroatom.
• a heteroaryl group is a group having 5 to 10 ring atoms (i.e., monocyclic, bicyclic or polycyclic), in some embodiments 5, 6, 9, or 10 ring atoms.
• a heteroaryl group has 6, 10, or 14 p electrons shared in a cyclic array; and having, in addition to carbon atoms, from one to five heteroatoms.
• Heteroaryl groups include, without limitation, thienyl, furanyl, pyrrolyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, oxadiazolyl, thiazolyl, isothiazolyl, thiadiazolyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, indolizinyl, purinyl, naphthyridinyl, and pteridinyl.
• a heteroaryl is a heterobiaryl group, such as bipyridyl and the like.
• the terms“heteroaryl” and“heteroar-”, as used herein, also include groups in which a heteroaromatic ring is fused to one or more aryl, cycloaliphatic, or heterocyclyl rings, where the radical or point of attachment is on the heteroaromatic ring.
• Non-limiting examples include indolyl, isoindolyl, benzothienyl, benzofuranyl, dibenzofuranyl, indazolyl, benzimidazolyl, benzthiazolyl, quinolyl, isoquinolyl, cinnolinyl, phthalazinyl, quinazolinyl, quinoxalinyl, 4 H quinolizinyl, carbazolyl, acridinyl, phenazinyl, phenothiazinyl, phenoxazinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, and pyrido[2,3-b]-l,4-oxazin-3(4H)-one.
• a heteroaryl group may be monocyclic, bicyclic or polycyclic.
• the term“heteroaryl” may be used interchangeably with the terms “heteroaryl ring,”“heteroaryl group,” or“heteroaromatic,” any of which terms include rings that are optionally substituted.
• the term“heteroaralkyl” refers to an alkyl group substituted by a heteroaryl group, wherein the alkyl and heteroaryl portions independently are optionally substituted.
• Heteroatom means an atom that is not carbon or hydrogen.
• a heteroatom is boron, oxygen, sulfur, nitrogen, phosphorus, or silicon (including oxidized forms of nitrogen, sulfur, phosphorus, or silicon; charged forms of nitrogen (e.g., quatemized forms, forms as in iminium groups, etc.), phosphorus, sulfur, oxygen; etc.).
• a heteroatom is oxygen, sulfur or nitrogen.
• Heterocycle As used herein, the terms“heterocycle,”“heterocyclyl,”“heterocyclic radical,” and“heterocyclic ring”, as used herein, are used interchangeably and refer to a monocyclic, bicyclic or polycyclic ring moiety (e.g., 3-30 membered) that is saturated or partially unsaturated and has one or more heteroatom ring atoms.
• a heterocyclyl group is a stable 5- to 7-membered monocyclic or 7- to 10-membered bicyclic heterocyclic moiety that is either saturated or partially unsaturated, and having, in addition to carbon atoms, one or more, preferably one to four, heteroatoms, as defined above.
• nitrogen When used in reference to a ring atom of a heterocycle, the term "nitrogen" includes substituted nitrogen.
• the nitrogen in a saturated or partially unsaturated ring having 0-3 heteroatoms selected from oxygen, sulfur and nitrogen, the nitrogen may be N (as in 3.4-dihydro-2// pyrrolyl). NH (as in pyrrolidinyl), or + NR (as in A substituted pyrrolidinyl).
• a heterocyclic ring can be attached to its pendant group at any heteroatom or carbon atom that results in a stable structure and any of the ring atoms can be optionally substituted.
• saturated or partially unsaturated heterocyclic radicals include, without limitation, tetrahydrofuranyl, tetrahydrothienyl, pyrrolidinyl, piperidinyl, pyrrolinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, decahydroquinolinyl, oxazolidinyl, piperazinyl, dioxanyl, dioxolanyl, diazepinyl, oxazepinyl, thiazepinyl, morpholinyl, and quinuclidinyl.
• heterocycle refers to an alkyl group substituted by a heterocyclyl, wherein the alkyl and heterocyclyl portions independently are optionally substituted.
• Internucleotidic linkage refers generally to a linkage linking nucleoside units of an oligonucleotide or a nucleic acid.
• an intemucleotidic linkage is a modified intemucleotidic linkage (not a natural phosphate linkage).
• an intemucleotidic linkage is a“modified intemucleotidic linkage” wherein at least one oxygen atom or -OH of a phosphodiester linkage is replaced by a different organic or inorganic moiety.
• a modified intemucleotidic linkage is a phosphorothioate linkage.
• an intemucleotidic linkage is one of, e.g., PNA (peptide nucleic acid) or PMO (phosphorodiamidate Morpholino oligomer) linkage.
• a modified intemucleotidic linkage is a non- negatively charged intemucleotidic linkage.
• a modified intemucleotidic linkage is a neutral intemucleotidic linkage (e.g., nOOl in certain provided oligonucleotides). It is understood by a person of ordinary skill in the art that an intemucleotidic linkage may exist as an anion or cation at a given pH due to the existence of acid or base moieties in the linkage.
• a modified intemucleotidic linkages is a modified intemucleotidic linkages designated as s, si, s2, s3, s4, s5, s6, s7, s8, s9, slO, si 1, sl2, sl3, sl4, sl5, sl6, sl7 and s 18 as described in WO 2017/210647.
• Linkage phosphorus as defined herein, the phrase“linkage phosphorus” is used to indicate that the particular phosphoms atom being referred to is the phosphoms atom present in the intemucleotidic linkage, which phosphoms atom corresponds to the phosphoms atom of a phosphodiester intemucleotidic linkage as occurs in naturally occurring DNA and RNA.
• a linkage phosphoms atom is in a modified intemucleotidic linkage, wherein each oxygen atom of a phosphodiester linkage is optionally and independently replaced by an organic or inorganic moiety.
• a linkage phosphoms atom is the P of, e.g., formula VII as defined herein. In some embodiments, a linkage phosphoms atom is chiral. In some embodiments, a linkage phosphoms atom is achiral (e.g., as in natural phosphate linkages).
• Modified nucleohase refer to a chemical moiety which is chemically distinct from a nucleobase, but which is capable of performing at least one function of a nucleobase.
• a modified nucleobase is a nucleobase which comprises a modification.
• a modified nucleobase is capable of at least one function of a nucleobase, e.g., forming a moiety in a polymer capable of base-pairing to a nucleic acid comprising an at least complementary sequence of bases.
• a modified nucleobase is substituted A, T, C, G, or U, or a substituted tautomer of A, T, C, G, or U.
• a modified nucleobase in the context of oligonucleotides refer to a nucleobase that is not A, T, C, G or U.
• Modified nucleoside refers to a moiety derived from or chemically similar to a natural nucleoside, but which comprises a chemical modification which differentiates it from a natural nucleoside.
• modified nucleosides include those which comprise a modification at the base and/or the sugar.
• modified nucleosides include those with a 2’ modification at a sugar.
• modified nucleosides also include abasic nucleosides (which lack a nucleobase).
• a modified nucleoside is capable of at least one function of a nucleoside, e.g., forming a moiety in a polymer capable of base-pairing to a nucleic acid comprising an at least complementary sequence of bases.
• Modified nucleotide includes any chemical moiety which differs structurally from a natural nucleotide but is capable of performing at least one function of a natural nucleotide.
• a modified nucleotide comprises a modification at a sugar, base and/or intemucleotidic linkage.
• a modified nucleotide comprises a modified sugar, modified nucleobase and/or modified intemucleotidic linkage.
• a modified nucleotide is capable of at least one function of a nucleotide, e.g., forming a subunit in a polymer capable of base-pairing to a nucleic acid comprising an at least complementary sequence of bases.
• Modified sugar refers to a moiety that can replace a sugar.
• a modified sugar mimics the spatial arrangement, electronic properties, or some other physicochemical property of a sugar.
• a modified sugar is substituted ribose or deoxyribose.
• a modified sugar comprises a 2’-modification. Examples of useful 2’-modification are widely utilized in the art and described herein.
• a 2’ -modification is 2’-OR, wherein R is optionally substituted C O aliphatic.
• a 2’ -modification is 2’-OMe.
• a 2’ -modification is 2’-MOE.
• a modified sugar is a bicyclic sugar (e.g., a sugar used in LNA, BNA, etc.).
• a modified sugar in the context of oligonucleotides, is a sugar that is not ribose or deoxyribose as typically found in natural RNA or DNA.
• Nucleobase refers to the parts of nucleic acids that are involved in the hydrogen-bonding that binds one nucleic acid strand to another complementary strand in a sequence specific manner.
• the most common naturally-occurring nucleobases are adenine (A), guanine (G), uracil (U), cytosine (C), and thymine (T).
• a naturally -occurring nucleobases are modified adenine, guanine, uracil, cytosine, or thymine.
• a naturally-occurring nucleobases are methylated adenine, guanine, uracil, cytosine, or thymine.
• a nucleobase comprises a heteroaryl ring wherein a ring atom is nitrogen, and when in a nucleoside, the nitrogen is bonded to a sugar moiety.
• a nucleobase comprises a heterocyclic ring wherein a ring atom is nitrogen, and when in a nucleoside, the nitrogen is bonded to a sugar moiety.
• a nucleobase is a“modified nucleobase,” a nucleobase other than adenine (A), guanine (G), uracil (U), cytosine (C), and thymine (T).
• a modified nucleobase is substituted A, T, C, G or U.
• a modified nucleobase is a substituted tautomer of A, T, C, G, or U.
• a modified nucleobases is methylated adenine, guanine, uracil, cytosine, or thymine.
• a modified nucleobase mimics the spatial arrangement, electronic properties, or some other physicochemical property of the nucleobase and retains the property of hydrogen-bonding that binds one nucleic acid strand to another in a sequence specific manner.
• a modified nucleobase can pair with all of the five naturally occurring bases (uracil, thymine, adenine, cytosine, or guanine) without substantially affecting the melting behavior, recognition by intracellular enzymes or activity of the oligonucleotide duplex.
• nucleobase also encompasses structural analogs used in lieu of natural or naturally-occurring nucleotides, such as modified nucleobases and nucleobase analogs.
• a nucleobase is optionally substituted A, T, C, G, or U, or an optionally substituted tautomer of A, T, C, G, or U.
• a“nucleobase” refers to a nucleobase unit in an oligonucleotide or a nucleic acid (e.g., A, T, C, G or U as in an oligonucleotide or a nucleic acid).
• Nucleoside refers to a moiety wherein a nucleobase or a modified nucleobase is covalently bound to a sugar or a modified sugar.
• a nucleoside is a natural nucleoside, e.g., adenosine, deoxyadenosine, guanosine, deoxyguanosine, thymidine, uridine, cytidine, or deoxycytidine.
• a nucleoside is a modified nucleoside, e.g., a substituted natural nucleoside selected from adenosine, deoxyadenosine, guanosine, deoxyguanosine, thymidine, uridine, cytidine, and deoxycytidine.
• a nucleoside is a modified nucleoside, e.g., a substituted tautomer of a natural nucleoside selected from adenosine, deoxyadenosine, guanosine, deoxyguanosine, thymidine, uridine, cytidine, and deoxycytidine.
• a “nucleoside” refers to a nucleoside unit in an oligonucleotide or a nucleic acid.
• nucleoside analog refers to a chemical moiety which is chemically distinct from a natural nucleoside, but which is capable of performing at least one function of a nucleoside.
• a nucleoside analog comprises an analog of a sugar and/or an analog of a nucleobase.
• a modified nucleoside is capable of at least one function of a nucleoside, e.g., forming a moiety in a polymer capable of base-pairing to a nucleic acid comprising a complementary sequence of bases.
• Nucleotide refers to a monomeric unit of a polynucleotide that consists of a nucleobase, a sugar, and one or more intemucleotidic linkages (e.g., phosphate linkages in natural DNA and R A).
• the naturally occurring bases [guanine, (G), adenine, (A), cytosine, (C), thymine, (T), and uracil (U)] are derivatives of purine or pyrimidine, though it should be understood that naturally and non-naturally occurring base analogs are also included.
• the naturally occurring sugar is the pentose (five-carbon sugar) deoxyribose (which forms DNA) or ribose (which forms RNA), though it should be understood that naturally and non-naturally occurring sugar analogs are also included.
• Nucleotides are linked via intemucleotidic linkages to form nucleic acids, or polynucleotides. Many intemucleotidic linkages are known in the art (such as, though not limited to, phosphate, phosphorothioates, boranophosphates and the like).
• a natural nucleotide comprises a naturally occurring base, sugar and intemucleotidic linkage.
• the term“nucleotide” also encompasses stmctural analogs used in lieu of natural or naturally- occurring nucleotides, such as modified nucleotides and nucleotide analogs.
• a “nucleotide” refers to a nucleotide unit in an oligonucleotide or a nucleic acid.
• Oligonucleotide refers to a polymer or oligomer of nucleotides, and may contain any combination of natural and non-natural nucleobases, sugars, and intemucleotidic linkages.
• Oligonucleotides can be single-stranded or double-stranded.
• a single-stranded oligonucleotide can have double-stranded regions (formed by two portions of the single-stranded oligonucleotide) and a double-stranded oligonucleotide, which comprises two oligonucleotide chains, can have single-stranded regions for example, at regions where the two oligonucleotide chains are not complementary to each other.
• Example oligonucleotides include, but are not limited to stmctural genes, genes including control and termination regions, self-replicating systems such as viral or plasmid DNA, single -stranded and double-stranded RNAi agents and other RNA interference reagents (RNAi agents or iRNA agents), shRNA, antisense oligonucleotides, ribozymes, microRNAs, microRNA mimics, supermirs, aptamers, antimirs, antagomirs, U1 adaptors, triplex-forming oligonucleotides, G-quadmplex oligonucleotides, RNA activators, immuno-stimulatory oligonucleotides, and decoy oligonucleotides.
• RNAi agents or iRNA agents RNA interference reagents
• shRNA antisense oligonucleotides
• ribozymes microRNAs
• microRNA mimics supermirs
• Oligonucleotides of the present disclosure can be of various lengths. In particular embodiments, oligonucleotides can range from about 2 to about 200 nucleosides in length. In various related embodiments, oligonucleotides, single-stranded, double-stranded, or triple-stranded, can range in length from about 4 to about 10 nucleosides, from about 10 to about 50 nucleosides, from about 20 to about 50 nucleosides, from about 15 to about 30 nucleosides, from about 20 to about 30 nucleosides in length. In some embodiments, the oligonucleotide is from about 9 to about 39 nucleosides in length.
• the oligonucleotide is at least 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 nucleosides in length. In some embodiments, the oligonucleotide is at least 4 nucleosides in length. In some embodiments, the oligonucleotide is at least 5 nucleosides in length. In some embodiments, the oligonucleotide is at least 6 nucleosides in length. In some embodiments, the oligonucleotide is at least 7 nucleosides in length. In some embodiments, the oligonucleotide is at least 8 nucleosides in length.
• the oligonucleotide is at least 9 nucleosides in length. In some embodiments, the oligonucleotide is at least 10 nucleosides in length. In some embodiments, the oligonucleotide is at least 11 nucleosides in length. In some embodiments, the oligonucleotide is at least 12 nucleosides in length. In some embodiments, the oligonucleotide is at least 15 nucleosides in length. In some embodiments, the oligonucleotide is at least 15 nucleosides in length. In some embodiments, the oligonucleotide is at least 16 nucleosides in length.
• the oligonucleotide is at least 17 nucleosides in length. In some embodiments, the oligonucleotide is at least 18 nucleosides in length. In some embodiments, the oligonucleotide is at least 19 nucleosides in length. In some embodiments, the oligonucleotide is at least 20 nucleosides in length. In some embodiments, the oligonucleotide is at least 25 nucleosides in length. In some embodiments, the oligonucleotide is at least 30 nucleosides in length. In some embodiments, the oligonucleotide is a duplex of complementary strands of at least 18 nucleosides in length.
• the oligonucleotide is a duplex of complementary strands of at least 21 nucleosides in length.
• each nucleoside counted in an oligonucleotide length independently comprises A, T, C, G, or U, or optionally substituted A, T, C, G, or U, or an optionally substituted tautomer of A, T, C, G or U.
• Oligonucleotide type is used to define an oligonucleotide that has a particular base sequence, pattern of backbone linkages (i.e.. pattern of intemucleotidic linkage types, for example, phosphate, phosphorothioate, etc.), pattern of backbone chiral centers (i.e. pattern of linkage phosphorus stereochemistry (/Zp/.S'p)). and pattern of backbone phosphorus modifications (e.g., pattern of “-X-L s -R 5 ” groups in formula VII).
• oligonucleotides of a common designated“type” are structurally, including stereochemically, identical to one another.
• each nucleotide unit of the oligonucleotide strand can be designed and/or selected in advance to have a particular stereochemistry at the linkage phosphorus and/or a particular modification at the linkage phosphorus, and/or a particular base, and/or a particular sugar.
• an oligonucleotide strand is designed and/or selected in advance to have a particular combination of stereocenters at the linkage phosphorus.
• an oligonucleotide strand is designed and/or determined to have a particular combination of modifications at the linkage phosphorus. In some embodiments, an oligonucleotide strand is designed and/or selected to have a particular combination of bases. In some embodiments, an oligonucleotide strand is designed and/or selected to have a particular combination of one or more of the above structural characteristics.
• the present disclosure provides compositions comprising or consisting of a plurality of oligonucleotide molecules (e.g., chirally controlled oligonucleotide compositions). In some embodiments, all such molecules are of the same type (i.e., are structurally identical to one another). In some embodiments, however, provided compositions comprise a plurality of oligonucleotides of different types, typically in pre -determined relative amounts.
• compounds, e.g., oligonucleotides, of the disclosure may contain optionally substituted and/or substituted moieties.
• substituted whether preceded by the term“optionally” or not, means that one or more hydrogens of the designated moiety are replaced with a suitable substituent.
• an“optionally substituted” group may have a suitable substituent at each substitutable position of the group, and when more than one position in any given structure may be substituted with more than one substituent selected from a specified group, the substituent may be either the same or different at every position.
• an optionally substituted group is unsubstituted.
• each R° may be substituted as defined herein and is independently hydrogen, Ci-20 aliphatic, Ci-20 heteroaliphatic having 1-5 heteroatoms independently selected from nitrogen, oxygen, sulfur, silicon and phosphorus, CH2 (Ce- 14 aryl), -0(CH 2 )o-i(C 6-i4 aryl), -CH 2 -(5-14 membered heteroaryl ring), a 5-20 membered, monocyclic, bicyclic, or polycyclic
• Suitable monovalent substituents on R° are independently halogen, -(CH2)o-2R*, - (haloR * ), -(CH 2 )O- 2 OH, -(CH 2 )O- 2 OR ⁇ , -(CH 2 )O-2CH(OR # ) 2 ; -0(haloR # ), -CN, -N 3 , -(CH 2 )o- 2 C(0)R # , - (CH 2 )O-2C(0)OH, -(CH 2 )O-2C(0)OR ⁇ , -(CH 2 )O-2SR # , -(CH 2 )O- 2 SH, -(CH 2 )O-2NH 2 , -(CH 2 )O-2NHR ⁇ , - (CH2)O-2NR # 2, -NO2, -SiR * 3 , -OSiR * 3 ,
• Suitable divalent substituents that are bound to vicinal substitutable carbons of an“optionally substituted” group include: -0(CR * 2)2-30-, wherein each independent occurrence of R * is selected from hydrogen, Ci_ 6 aliphatic which may be substituted as defined below, and an unsubstituted 5-6-membered saturated, partially unsaturated, and aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur.
• Suitable substituents on the aliphatic group of R * are independently halogen, -R # , -(haloR*), -OH, -OR*, -0(haloR # ), -CN, -C(0)OH, -C(0)OR # , -NH 2 , NHR ⁇ -NRV or -NO2, wherein each R* is unsubstituted or where preceded by“halo” is substituted only with one or more halogens, and is independently C 1-4 aliphatic, -CH2PI1, -0(03 ⁇ 4)o-iR1i, or a 5-6-membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur.
• suitable substituents on a substitutable nitrogen are independently -R ⁇ , -NR ⁇ 2 , -C(0)R ⁇ , -C(0)OR ⁇ , -C(0)C(0)R ⁇ , -C(0)CH 2 C(0)R ⁇ , -S(0) 2 R ⁇ , -S(0) 2 NR ⁇ 2 , -C(S)NR ⁇ 2 , - C(NH)NR ⁇ 2 , or -N(R ⁇ )S(0) 2 R ⁇ ; wherein each R ⁇ is independently hydrogen, Ci_ 6 aliphatic which may be substituted as defined below, unsubstituted -OPh, or an unsubstituted 5-6-membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur, or, notwithstanding the definition above, two independent occurrences of R ⁇ , taken together with their intervening atom(s) form
• Suitable substituents on the aliphatic group of R ⁇ are independently halogen, -R # , -(haloR*), -OH, -OR*, -0(haloR # ), -CN, -C(0)OH, -C(0)OR # , -NH 2 , NHR ⁇ -NRV or -N0 2 , wherein each R* is unsubstituted or where preceded by“halo” is substituted only with one or more halogens, and is independently Ci- 4 aliphatic, -CH 2 Ph, -0(CH 2 )o-iPh, or a 5-6-membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur.
• Partially unsaturated refers to a ring moiety that includes at least one double or triple bond.
• the term“partially unsaturated” is intended to encompass rings having multiple sites of unsaturation, but is not intended to include aryl or heteroaryl moieties, as herein defined.
• composition refers to an active agent, formulated together with one or more pharmaceutically acceptable carriers.
• an active agent is present in unit dose amount appropriate for administration in a therapeutic regimen that shows a statistically significant probability of achieving a predetermined therapeutic effect when administered to a relevant population.
• pharmaceutical compositions may be specially formulated for administration in solid or liquid form, including those adapted for the following: oral administration, for example, drenches (aqueous or non-aqueous solutions or suspensions), tablets, e.g.
• parenteral administration for example, by subcutaneous, intramuscular, intravenous or epidural injection as, for example, a sterile solution or suspension, or sustained-release formulation
• parenteral application for example, as a cream, ointment, or a controlled-release patch or spray applied to the skin, lungs, or oral cavity
• intravaginally or intrarectally for example, as a pessary, cream, or foam
• sublingually ocularly
• transdermally or nasally, pulmonary, and to other mucosal surfaces.
• compositions and/or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit/risk ratio.
• composition or vehicle such as a liquid or solid fdler, diluent, excipient, or solvent encapsulating material, involved in carrying or transporting the subject compound from one organ, or portion of the body, to another organ, or portion of the body.
• Each carrier must be“acceptable” in the sense of being compatible with the other ingredients of the formulation and not injurious to the patient.
• materials which can serve as pharmaceutically-acceptable carriers include: sugars, such as lactose, glucose and sucrose; starches, such as com starch and potato starch; cellulose, and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; powdered tragacanth; malt; gelatin; talc; excipients, such as cocoa butter and suppository waxes; oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, com oil and soybean oil; glycols, such as propylene glycol; polyols, such as glycerin, sorbitol, mannitol and polyethylene glycol; esters, such as ethyl oleate and ethyl laurate; agar; buffering agents, such as magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen-free water; isotonic saline
• compositions that are appropriate for use in pharmaceutical contexts, i. e.. salts which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and lower animals without undue toxicity, irritation, allergic response and the like, and are commensurate with a reasonable benefit/risk ratio.
• Pharmaceutically acceptable salts are well known in the art. For example, S. M. Berge, et al. describes pharmaceutically acceptable salts in detail in J. Pharmaceutical Sciences, 66: 1-19 (1977).
• pharmaceutically acceptable salt include, but are not limited to, nontoxic acid addition salts, which are salts of an amino group formed with inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid and perchloric acid or with organic acids such as acetic acid, maleic acid, tartaric acid, citric acid, succinic acid or malonic acid or by using other methods used in the art such as ion exchange.
• nontoxic acid addition salts which are salts of an amino group formed with inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid and perchloric acid or with organic acids such as acetic acid, maleic acid, tartaric acid, citric acid, succinic acid or malonic acid or by using other methods used in the art such as ion exchange.
• pharmaceutically acceptable salts include, but are not limited to, adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate, gluconate, hemisulfate, heptanoate, hexanoate, hydroiodide, 2-hydroxy- ethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methane sulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate,
• a provided compound comprises one or more acidic groups, e.g., an oligonucleotide, and a pharmaceutically acceptable salt is an alkali, alkaline earth metal, or ammonium (e.g., an ammonium salt of N(R) 3 , wherein each R is independently defined and described in the present disclosure) salt.
• a pharmaceutically acceptable salt is a sodium salt.
• a pharmaceutically acceptable salt is a potassium salt.
• a pharmaceutically acceptable salt is a calcium salt.
• pharmaceutically acceptable salts include, when appropriate, nontoxic ammonium, quaternary ammonium, and amine cations formed using counterions such as halide, hydroxide, carboxylate, sulfate, phosphate, nitrate, alkyl having from 1 to 6 carbon atoms, sulfonate and aryl sulfonate.
• a provided compound comprises more than one acid groups, for example, an oligonucleotide may comprise two or more acidic groups (e.g., in natural phosphate linkages and/or modified intemucleotidic linkages).
• a pharmaceutically acceptable salt, or generally a salt, of such a compound comprises two or more cations, which can be the same or different.
• all ionizable hydrogen e.g., in an aqueous solution with a pKa no more than about 11, 10, 9, 8, 7, 6, 5, 4, 3, or 2; in some embodiments, no more than about 7; in some embodiments, no more than about 6; in some embodiments, no more than about 5; in some embodiments, no more than about 4; in some embodiments, no more than about 3 in the acidic groups are replaced with cations.
• each phosphorothioate and phosphate group independently exists in its salt form (e.g., if sodium salt, -0-P(0)(SNa)-0- and -0-P(0)(0Na)-0-, respectively).
• each phosphorothioate and phosphate intemucleotidic linkage independently exists in its salt form (e.g., if sodium salt, -0-P(0)(SNa)-0- and -0-P(0)(0Na)-0-, respectively).
• a pharmaceutically acceptable salt is a sodium salt of an oligonucleotide.
• a pharmaceutically acceptable salt is a sodium salt of an oligonucleotide, wherein each acidic phosphate and modified phosphate group (e.g., phosphorothioate, phosphate, etc.), if any, exists as a salt form (all sodium salt).
• each acidic phosphate and modified phosphate group e.g., phosphorothioate, phosphate, etc.
• predetermined By predetermined (or pre-determined) is meant deliberately selected or non- random or controlled, for example as opposed to randomly occurring, random, or achieved without control.
• predetermined By reading the present specification, will appreciate that the present disclosure provides technologies that permit selection of particular chemistry and/or stereochemistry features to be incorporated into oligonucleotide compositions, and further permits controlled preparation of oligonucleotide compositions having such chemistry and/or stereochemistry features.
• Such provided compositions are“predetermined” as described herein. Compositions that may contain certain oligonucleotides because they happen to have been generated through a process that are not controlled to intentionally generate the particular chemistry and/or stereochemistry features are not “predetermined” compositions.
• a predetermined composition is one that can be intentionally reproduced (e.g., through repetition of a controlled process).
• a predetermined level of a plurality of oligonucleotides in a composition means that the absolute amount, and/or the relative amount (ratio, percentage, etc.) of the plurality of oligonucleotides in the composition is controlled.
• a predetermined level of a plurality of oligonucleotides in a composition is achieved through chirally controlled oligonucleotide preparation.
• Protecting group The term“protecting group,” as used herein, is well known in the art and includes those described in detail in Protecting Groups in Organic Synthesis, T. W. Greene and P. G. M. Wuts, 3 rd edition, John Wiley & Sons, 1999, the entirety of which is incorporated herein by reference. Also included are those protecting groups specially adapted for nucleoside and nucleotide chemistry described in Current Protocols in Nucleic Acid Chemistry, edited by Serge L. Beaucage et al. 06/2012, the entirety of Chapter 2 is incorporated herein by reference.
• Suitable amino-protecting groups include methyl carbamate, ethyl carbamante, 9-fluorenylmethyl carbamate (Fmoc), 9-(2-sulfo)fluorenylmethyl carbamate, 9-(2,7-dibromo)fluoroenylmethyl carbamate, 2.7-di-/-butyl-
• o-nitophenylacetamide o-nitrophenoxyacetamide, acetoacetamide, (A -dithiobenzyloxycarbonylamino)acetamide.
• A- 1.1 ,4,4-tetramethyldisilylazacyclopentane adduct STABASE
• 5-substituted l,3-dimethyl-l,3,5-triazacyclohexan-2-one 5-substituted 1,3— dibenzyl-1, 3, 5-triazacyclohexan-2-one, 1-substituted 3,5-dinitro-4-pyridone, A'-mcthylaminc.
• A- 1. 1- dimethylthiomethyleneamine, A'-b c n z y 1 i dc n c am i n c .
• Dpp diphenylphosphinamide
• Mpt dimethylthiophosphinamide
• Ppt diphenylthiophosphinamide
• dialkyl phosphoramidates dibenzyl phosphoramidate, diphenyl phosphoramidate
• benzenesulfenamide o-nitrobenzenesulfenamide (Nps)
• 2,4- dinitrobenzenesulfenamide pentachlorobenzenesulfenamide, 2-nitro-4-methoxybenzenesulfenamide, triphenylmethylsulfenamide, 3-nitropyridinesulfenamide (Npys), / -to 1 uc n c s ul fo n am i dc (Ts), benzenesulfonamide, 2,3,6,-trimethyl-4-methoxybenzenesulfonamide (Mtr), 2,4,6- trimethoxybenzenesulfonamide (Mt
• Suitably protected carboxylic acids further include, but are not limited to, silyl— , alkyl-, alkenyl-, aryl-, and arylalkyl-protected carboxylic acids.
• suitable silyl groups include trimethylsilyl, triethylsilyl, t-butyldimethylsilyl, t-butyldiphenylsilyl, triisopropylsilyl, and the like.
• suitable alkyl groups include methyl, benzyl, p-methoxybenzyl, 3,4-dimethoxybenzyl, trityl, t-butyl, tetrahydropyran-2-yl.
• suitable alkenyl groups include allyl.
• suitable aryl groups include optionally substituted phenyl, biphenyl, or naphthyl.
• suitable arylalkyl groups include optionally substituted benzyl (e.g., p-methoxybenzyl (MPM), 3,4- dimethoxybenzyl, O-nitrobenzyl, p-nitrobenzyl, p-halobenzyl, 2,6-dichlorobenzyl, p-cyanobenzyl), and 2- and 4-picolyl.
• Suitable hydroxyl protecting groups include methyl, methoxylmethyl (MOM), methylthiomethyl (MTM), /-butylthiomethyl.
• MOM methoxylmethyl
• MTM methylthiomethyl
• PMBM benzyloxymethyl
• GUM guaiacolmethyl
• the protecting groups include methylene acetal, ethylidene acetal, 1 -t- butylethylidene ketal, 1-phenylethylidene ketal, (4-methoxyphenyl)ethylidene acetal, 2,2,2- trichloroethylidene acetal, acetonide, cyclopentylidene ketal, cyclohexylidene ketal, cycloheptylidene ketal, benzylidene acetal, / -methoxybenzylidene acetal, 2,4-dimethoxybenzylidene ketal, 3,4- dimethoxybenzybdene acetal, 2-nitrobenzylidene acetal, methoxymethylene acetal, ethoxymethylene acetal, dimethoxymethylene ortho ester, 1-methoxy ethyliden
• a hydroxyl protecting group is acetyl, t-butyl, tbutoxymethyl, methoxymethyl, tetrahydropyranyl, 1 -ethoxyethyl, 1 -(2-chloroethoxy)ethyl, 2- trimethylsilylethyl, p- chlorophenyl, 2,4-dinitrophenyl, benzyl, benzoyl, p-phenylbenzoyl, 2,6- dichlorobenzyl, diphenylmethyl, p-nitrobenzyl, triphenylmethyl (trityl), 4,4'-dimethoxytrityl, trimethylsilyl, triethylsilyl, t- butyldimethylsilyl, t-butyldiphenylsilyl, triphenylsilyl, triisopropylsilyl, benzoylformate, chloroacetyl, trichlor
• each of the hydroxyl protecting groups is, independently selected from acetyl, benzyl, t- butyldimethylsilyl, t-butyldiphenylsilyl and 4,4'- dimethoxytrityl.
• the hydroxyl protecting group is selected from the group consisting of trityl, monomethoxytrityl and 4,4'-dimethoxytrityl group.
• a phosphorous linkage protecting group is a group attached to the phosphorous linkage (e.g., an intemucleotidic linkage) throughout oligonucleotide synthesis.
• a protecting group is attached to a sulfur atom of an phosphorothioate group. In some embodiments, a protecting group is attached to an oxygen atom of an intemucleotide phosphorothioate linkage. In some embodiments, a protecting group is attached to an oxygen atom of the intemucleotide phosphate linkage.
• a protecting group is 2-cyanoethyl (CE or Cne), 2-trimethylsilylethyl, 2-nitroethyl, 2- sulfonylethyl, methyl, benzyl, o-nitrobenzyl, 2-(p-nitrophenyl)ethyl (NPE or Npe), 2-phenylethyl, 3-(N- tert-butylcarboxamido)- 1 -propyl, 4-oxopentyl, 4-methylthio-l -butyl, 2-cyano-l,l-dimethylethyl, 4 -N- methylaminobutyl, 3-(2-pyridyl)-l-propyl, 2-
• the term“substantially” refers to the qualitative condition of exhibiting total or near-total extent or degree of a characteristic or property of interest.
• a base sequence which is substantially complementary to a second sequence is not identical to the second sequence, but is mostly or nearly identical to the second sequence.
• the term “substantially” is therefore used herein to capture the potential lack of completeness inherent in many biological and/or chemical phenomena.
• sugar refers to a monosaccharide or polysaccharide in closed and/or open form.
• sugars are monosaccharides.
• sugars are polysaccharides.
• Sugars include, but are not limited to, ribose, deoxyribose, pentofuranose, pentopyranose, and hexopyranose moieties.
• the term“sugar” also encompasses structural analogs used in lieu of conventional sugar molecules, such as glycol, polymer of which forms the backbone of the nucleic acid analog, glycol nucleic acid (“GNA”), etc.
• a sugar is a R A or DNA sugar (ribose or deoxyribose).
• a sugar is a modified ribose or deoxyribose sugar, e.g., 2’-modified, 5’-modified, etc.
• modified sugars when used in oligonucleotides and/or nucleic acids, modified sugars may provide one or more desired properties, activities, etc.
• a sugar is optionally substituted ribose or deoxyribose.
• a“sugar” refers to a sugar unit in an oligonucleotide or a nucleic acid.
• Susceptible to An individual who is“susceptible to” a disease, disorder and/or condition is one who has a higher risk of developing the disease, disorder and/or condition than does a member of the general public. In some embodiments, an individual who is susceptible to a disease, disorder and/or condition is predisposed to have that disease, disorder and/or condition. In some embodiments, an individual who is susceptible to a disease, disorder and/or condition may not have been diagnosed with the disease, disorder and/or condition. In some embodiments, an individual who is susceptible to a disease, disorder and/or condition may exhibit symptoms of the disease, disorder and/or condition.
• an individual who is susceptible to a disease, disorder and/or condition may not exhibit symptoms of the disease, disorder and/or condition. In some embodiments, an individual who is susceptible to a disease, disorder, and/or condition will develop the disease, disorder, and/or condition. In some embodiments, an individual who is susceptible to a disease, disorder, and/or condition will not develop the disease, disorder, and/or condition.
• therapeutic agent in general refers to any agent that elicits a desired effect (e.g., a desired biological, clinical, or pharmacological effect) when administered to a subject.
• a desired effect e.g., a desired biological, clinical, or pharmacological effect
• an agent is considered to be a therapeutic agent if it demonstrates a statistically significant effect across an appropriate population.
• an appropriate population is a population of subjects suffering from and/or susceptible to a disease, disorder or condition.
• an appropriate population is a population of model organisms.
• an appropriate population may be defined by one or more criterion such as age group, gender, genetic background, preexisting clinical conditions, prior exposure to therapy.
• a therapeutic agent is a substance that alleviates, ameliorates, relieves, inhibits, prevents, delays onset of, reduces severity of, and/or reduces incidence of one or more symptoms or features of a disease, disorder, and/or condition in a subject when administered to the subject in an effective amount.
• a“therapeutic agent” is an agent that has been or is required to be approved by a government agency before it can be marketed for administration to humans.
• a “therapeutic agent” is an agent for which a medical prescription is required for administration to humans.
• a therapeutic agent is a provided compound, e.g. , a provided oligonucleotide.
• therapeutically effective amount means an amount of a substance (e.g., a therapeutic agent, composition, and/or formulation) that elicits a desired biological response when administered as part of a therapeutic regimen.
• a therapeutically effective amount of a substance is an amount that is sufficient, when administered to a subject suffering from or susceptible to a disease, disorder, and/or condition, to treat, diagnose, prevent, and/or delay the onset of the disease, disorder, and/or condition.
• the effective amount of a substance may vary depending on such factors as the desired biological endpoint, the substance to be delivered, the target cell or tissue, etc.
• the effective amount of compound in a formulation to treat a disease, disorder, and/or condition is the amount that alleviates, ameliorates, relieves, inhibits, prevents, delays onset of, reduces severity of and/or reduces incidence of one or more symptoms or features of the disease, disorder, and/or condition.
• a therapeutically effective amount is administered in a single dose; in some embodiments, multiple unit doses are required to deliver a therapeutically effective amount.
• Treat refers to any method used to partially or completely alleviate, ameliorate, relieve, inhibit, prevent, delay onset of, reduce severity of, and/or reduce incidence of one or more symptoms or features of a disease, disorder, and/or condition.
• Treatment may be administered to a subject who does not exhibit signs of a disease, disorder, and/or condition.
• treatment may be administered to a subject who exhibits only early signs of the disease, disorder, and/or condition, for example for the purpose of decreasing the risk of developing pathology associated with the disease, disorder, and/or condition.
• Unsaturated means that a moiety has one or more units of unsaturation.
• the present disclosure provides technologies for preparing
• oligonucleotide compositions particularly chirally controlled oligonucleotide compositions, with unexpectedly improved crude purity and yield.
• provided technologies can dramatically reduce costs of goods, and in some embodiments, enable large production of therapeutic oligonucleotides at commercially acceptable conditions, e.g., cost, purity, yield, etc., for clinical uses and commercialization.
• provided technologies enable production of compositions of various oligonucleotides, independent of base sequences, chemical/stereochemical modifications, modes of activities, chiral auxiliaries, etc. Example embodiments of provided technologies are described herein.
• oligonucleotide compositions of provided technologies e.g., product oligonucleotide compositions of various steps, final oligonucleotide compositions, etc., are chirally controlled oligonucleotide composition.
• each oligonucleotide of a plurality of oligonucleotides is independently an oligonucleotide of formula O-I or a salt thereof.
• an oligonucleotide is of formula O-I or a salt thereof:
• R 5S is independently R’ or -OR’ ;
• each L s is independently a covalent bond, or a bivalent, optionally substituted, linear or branched group selected from a Ci-30 aliphatic group and a Ci-30 heteroabphatic group having 1-10 heteroatoms, wherein one or more methylene units are optionally and independently replaced by an optionally substituted group selected from Ci- 6 alkylene, Ci- 6 alkenylene, ⁇ CoC ⁇ , a bivalent C1-G5 heteroabphatic group having 1-5 heteroatoms, -C(R’)2-, -Cy-, -0-, -S-, -S-S-, -N(R’)-, -C(O)-, -C(S)-,
• each -Cy- is independently an optionally substituted bivalent group selected from a C3-20 cycloaliphatic ring, a Co- 20 aryl ring, a 5-20 membered heteroaryl ring having 1-10 heteroatoms, and a 3- 20 membered heterocyclyl ring having 1-10 heteroatoms ;
• each Cy L is independently an optionally substituted tetravalent group selected from a C3-20 cycloaliphatic ring, a Co- 20 aryl ring, a 5-20 membered heteroaryl ring having 1-10 heteroatoms, and a 3- 20 membered heterocyclyl ring having 1-10 heteroatoms;
• each Ring A s is independently an optionally substituted 3-20 membered monocyclic, bicyclic or polycyclic ring having 0-10 heteroatoms;
• each R s is independently -H, halogen, -CN, -N3, -NO, -NO2, -L s -R’, -L s -Si(R') 3 , -L s -OR’, -L S -SR’, -L S -N(R’) 2 , -0-L S -R’, -0-L s -Si(R) 3 , -0-L s -OR’, -0-L s -SR’, or -0-L s -N(R’) 2 ;
• each t is independently 0-20;
• each BA is independently an optionally substituted group selected from C3-30 cycloaliphatic, Co- 30 aryl, C5-30 heteroaryl having 1-10 heteroatoms, C3-30 heterocyclyl having 1-10 heteroatoms, a natural nucleobase moiety, and a modified nucleobase moiety;
• each L p is independently an intemucleotidic linkage
• z is 1-1000
• L 3E is -L s - or -L s -L s -;
• R 3E is -R’, -L s -R’, -OR’, or a support
• each R’ is independently -R, -C(0)R, -C(0)OR, or -S(0) 2 R;
• each R is independently -H, or an optionally substituted group selected from Ci-30 aliphatic, Ci-30 heteroaliphatic having 1-10 heteroatoms, 0,- 30 aryl, Co-30 arylaliphatic, Co-30 arylheteroaliphatic having 1- 10 heteroatoms, 5-30 membered heteroaryl having 1-10 heteroatoms, and 3-30 membered heterocyclyl having 1-10 heteroatoms, or
• two or more R groups on the same atom are optionally and independently taken together with the atom to form an optionally substituted, 3-30 membered, monocyclic, bicyclic or polycyclic ring having, in addition to the atom, 0-10 heteroatoms; or
• two or more R groups on two or more atoms are optionally and independently taken together with their intervening atoms to form an optionally substituted, 3-30 membered, monocyclic, bicyclic or polycyclic ring having, in addition to the intervening atoms, 0-10 heteroatoms.
• an oligonucleotide composition is a chirally controlled oligonucleotide composition.
• a chirally controlled oligonucleotide composition is an oligonucleotide composition comprising a plurality of oligonucleotides which share:
• composition which composition is chirally controlled in that level of the plurality of oligonucleotides in the composition is predetermined.
• a chirally controlled oligonucleotide composition is an oligonucleotide composition comprising a plurality of oligonucleotides, wherein oligonucleotides of the plurality are of a particular oligonucleotide type defined by:
• composition which composition is chirally controlled in that level of the plurality of oligonucleotides in the composition is predetermined.
• a chirally controlled oligonucleotide composition is an oligonucleotide composition comprising a plurality of oligonucleotides which share:
• composition is a substantially pure preparation of a single oligonucleotide in that a predetermined level of the oligonucleotides in the composition have the common base sequence and length, the common pattern of backbone linkages, and the common pattern of backbone chiral centers.
• a chirally controlled oligonucleotide composition is an oligonucleotide composition comprising a plurality of oligonucleotides, wherein:
• oligonucleotides of the plurality share the same base sequence
• oligonucleotides of the plurality share the same pattern of backbone linkages
• oligonucleotides of the plurality comprise at least one chirally controlled intemucleotidic linkage, which intemucleotidic linkage is chirally controlled in that oligonucleotides of the plurality share the same stereochemical configuration at the chiral linkage phosphorus of the intemucleotidic linkage;
• oligonucleotides sharing the same base sequence in the composition are oligonucleotides of the plurality, wherein DS is at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%, and Nc is the number of chirally controlled intemucleotidic linkage.
• a chirally controlled oligonucleotide composition is an oligonucleotide composition comprising a plurality of oligonucleotides, wherein:
• oligonucleotides of the plurality share the same constitution
• oligonucleotides of the plurality comprise at least one chirally controlled intemucleotidic linkage, which intemucleotidic linkage is chirally controlled in that oligonucleotides of the plurality share the same stereochemical configuration at the chiral linkage phosphorus of the intemucleotidic linkage; and wherein no less than ((DS) Nc * 100)% of all oligonucleotides sharing the same base sequence in the composition are oligonucleotides of the plurality, wherein DS is at least 80%, 85%, 90%, 91%, 92%,
• Nc is the number of chirally controlled intemucleotidic linkage.
• provided oligonucleotides comprise 1-30 non-natural intemucleotidic linkages (not -0-P(0)(0H)-0- or salt forms thereof). In some embodiments, provided oligonucleotides comprise 2-30 non-natural intemucleotidic linkages. In some embodiments, provided oligonucleotides comprise 5-30 non-natural intemucleotidic linkages. In some embodiments, provided oligonucleotides comprise 10-30 non-natural intemucleotidic linkages.
• provided oligonucleotides comprise 1-30 chirally controlled intemucleotidic linkages. In some embodiments, provided oligonucleotides comprise 2-30 chirally controlled intemucleotidic linkages. In some embodiments, provided oligonucleotides comprise 5-30 chirally controlled intemucleotidic linkages. In some embodiments, provided oligonucleotides comprise 10-30 chirally controlled intemucleotidic linkages. In some embodiments, provided oligonucleotides comprise 1 chirally controlled intemucleotidic linkage.
• provided oligonucleotides comprise 2 chirally controlled intemucleotidic linkages. In some embodiments, provided oligonucleotides comprise 3 chirally controlled intemucleotidic linkages. In some embodiments, provided oligonucleotides comprise 4 chirally controlled intemucleotidic linkages. In some embodiments, provided oligonucleotides comprise 5 chirally controlled intemucleotidic linkages. In some embodiments, provided oligonucleotides comprise 6 chirally controlled intemucleotidic linkages. In some embodiments, provided oligonucleotides comprise 7 chirally controlled intemucleotidic linkages.
• provided oligonucleotides comprise 8 chirally controlled intemucleotidic linkages. In some embodiments, provided oligonucleotides comprise 9 chirally controlled intemucleotidic linkages. In some embodiments, provided oligonucleotides comprise 10 chirally controlled intemucleotidic linkages. In some embodiments, provided oligonucleotides comprise 11 chirally controlled intemucleotidic linkages. In some embodiments, provided oligonucleotides comprise 12 chirally controlled intemucleotidic linkages. In some embodiments, provided oligonucleotides comprise 13 chirally controlled intemucleotidic linkages. In some embodiments, provided oligonucleotides comprise
• provided oligonucleotides have
• provided oligonucleotides have
• provided oligonucleotides have
• provided oligonucleotides have
• provided oligonucleotides have
• provided oligonucleotides have
• chirally controlled intemucleotidic linkages In some embodiments, about 1-100% of all intemucleotidic linkages are chirally controlled intemucleotidic linkages. In some embodiments, about 1- 100% of all chiral intemucleotidic linkages (comprising chiral linkage phosphoms) are chirally controlled intemucleotidic linkages. In some embodiments, a percentage is about 5%-100%.
• a percentage is at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99%. In some embodiments, a percentage is about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99%. In some embodiments, each chiral intemucleotidic linkage is chirally controlled.
• a portion of or all of chirally controlled intemucleotidic linkages in provided oligonucleotides or one or more segments thereof are consecutive.
• all chirally controlled intemucleotidic linkages in provided oligonucleotides or one or more segments thereof are consecutive.
• provided oligonucleotides comprise 1-30 natural phosphate linkages (not -0-P(0)(0H)-0- or salt forms thereof). In some embodiments, provided oligonucleotides, in addition to natural phosphate linkages, or chiral intemucleotidic linkages, or chirally controlled intemucleotidic linkages as described herein, further comprise 1-30 natural phosphate linkages (not -0-P(0)(0H)-0- or salt forms thereof). In some embodiments, provided oligonucleotides comprise 2- 30 natural phosphate linkages. In some embodiments, provided oligonucleotides comprise 5-30 natural phosphate linkages.
• provided oligonucleotides comprise 10-30 natural phosphate linkages. In some embodiments, provided oligonucleotides comprise 1 chirally controlled intemucleotidic linkage. In some embodiments, provided oligonucleotides comprise 2 natural phosphate linkages. In some embodiments, provided oligonucleotides comprise 3 natural phosphate linkages. In some embodiments, provided oligonucleotides comprise 4 natural phosphate linkages. In some embodiments, provided oligonucleotides comprise 5 natural phosphate linkages. In some embodiments, provided oligonucleotides comprise 6 natural phosphate linkages.
• provided oligonucleotides comprise 7 natural phosphate linkages. In some embodiments, provided oligonucleotides comprise 8 natural phosphate linkages In some embodiments, provided oligonucleotides comprise 9 natural phosphate linkages In some embodiments, provided oligonucleotides comprise 10 natural phosphate linkages In some embodiments, provided oligonucleotides comprise 11 natural phosphate linkages In some embodiments, provided oligonucleotides comprise 12 natural phosphate linkages In some embodiments, provided oligonucleotides comprise 13 natural phosphate linkages In some embodiments, provided oligonucleotides comprise 14 natural phosphate linkages In some embodiments, provided oligonucleotides have 15 natural phosphate linkages.
• provided oligonucleotides have 16 natural phosphate linkages. In some embodiments, provided oligonucleotides have 17 natural phosphate linkages. In some embodiments, provided oligonucleotides have 18 natural phosphate linkages. In some embodiments, provided oligonucleotides have 19 natural phosphate linkages. In some embodiments, provided oligonucleotides have 20 natural phosphate linkages. In some embodiments, about 1-100% of all intemucleotidic linkages are natural phosphate linkages.
• intemucleotidic linkages are natural phosphate linkages
• about 1-99% of all intemucleotidic linkages are non-natural intemucleotidic linkages (e.g., intemucleotidic linkages of formula VII, VII-a-1, VII-a-2, Vll-b, VII-c, Vll-d, Vll-e, NL-n-1, NL-n-2, NL-n-3, NL-n-4, NL, NL- a-1, NL-a-2, NL-b-1, NL-b-2, NL-c-1, NL-c-2, NL-d-1, or NL-d-2, or a salt form thereof and are non natural intemucleotidic linkages).
• intemucleotidic linkages are natural phosphate linkages
• about 1-99% of all intemucleotidic linkages are chiral intemucleotidic linkages (e.g., intemucleotidic linkages of formula VII, VII-a-1, VII-a-2, Vll-b, VII-c, Vll-d, Vll-e, NL-n-1, NL-n-2, NL-n-3, NL-n-4, NL, NL-a-1, NL-a-2, NL-b-1, NL-b-2, NL-c-1, NL-c-2, NL-d-1, or NL-d-2, or a salt form thereof and are chiral).
• a percentage is about 5%-100%. In some embodiments, a percentage is at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99%. In some embodiments, a percentage is about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99%.
• each non-natural intemucleotidic linkage independently has the structure of formula VII, VII-a-1, VII-a-2, Vll-b, VII-c, Vll-d, Vll-e, NL-n-1, NL-n-2, NL-n- 3, NL-n-4, NL, NL-a-1, NL-a-2, NL-b-1, NL-b-2, NL-c-1, NL-c-2, NL-d-1, or NL-d-2, or a salt form thereof, wherein the structure of formula VII, VII-a-1, VII-a-2, Vll-b, VII-c, Vll-d, Vll-e, NL-n-1, NL- n-2, NL-n-3, NL-n-4, NL, NL-a-1, NL-a-2, NL-b-1, NL-b-2, NL-c-1, NL-c-2, NL-d-1, or NL-d-2, or a salt form thereof
• each chiral linkage independently has the structure of formula VII, VII-a-1, VII- a-2, Vll-b, VII-c, Vll-d, Vll-e, NL-n-1, NL-n-2, NL-n-3, NL-n-4, NL, NL-a-1, NL-a-2, NL-b-1, NL- b-2, NL-c-1, NL-c-2, NL-d-1, or NL-d-2, or a salt form thereof, wherein the stmcture of formula VII, VII-a-1, VII-a-2, Vll-b, VII-c, Vll-d, Vll-e, NL-n-1, NL-n-2, NL-n-3, NL-n-4, NL, NL-a-1, NL-a-2, NL-b-1, NL-b-2, NL-c-1, NL-c-2, NL-d-1, or NL-d-2, or a salt form thereof, where
• each chirally controlled phosphate linkage independently has the stmcture of formula VII, VII-a-1, VII-a-2, Vll-b, VII-c, Vll-d, Vll-e, NL-n-1, NL-n-2, NL-n-3, NL-n-4, NL, NL-a-1, NL-a-2, NL-b-1, NL-b-2, NL-c-1, NL-c-2, NL- d-1, or NL-d-2, or a salt form thereof, wherein the stmcture of formula VII, VII-a-1, VII-a-2, Vll-b, VII-c, Vll-d, Vll-e, NL-n-1, NL-n-2, NL-n-3, NL-n-4, NL, NL-a-1, NL-a-2, NL-b-1, NL-b-2, NL-c-1, NL-c-2, NL-d-1, or NL-d-2,
• a portion of or all of natural phosphate linkages in provided oligonucleotides or one or more segments thereof are consecutive.
• all natural phosphate linkages in provided oligonucleotides or one or more segments thereof are consecutive.
• a non-natural intemucleotidic linkage is a phosphorothioate linkage or a salt form thereof (-0-P(0)(SH)-0- or a salt form thereof).
• a chiral intemucleotidic linkage is a phosphorothioate linkage or a salt form thereof (-0-P(0)(SH)-0- or a salt form thereof).
• a chirally controlled intemucleotidic linkage is a phosphorothioate linkage or a salt form thereof (-0-P(0)(SH)-0- or a salt form thereof).
• provided oligonucleotides comprise 5-200, 5-150, 5-100, 5-50, 5-40, 5-35, 5-30, 5-25, 10-200, 10-150, 10-100, 10-50, 10-40, 10-35, 10-30, 10-25, 15-200, 15-150, 15-100, 15- 50, 15-40, 15-35, 15-30, or 15-25 nucleobases.
• provided oligonucleotides comprise at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 35, 40, 45, or 50 nucleobases.
• provided oligonucleotides comprise at least 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 35, 40,
• provided oligonucleotides comprise at least 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 35, 40, 45, or 50 nucleobases. In some embodiments, provided oligonucleotides comprise at least 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 35, 40, 45, or 50 nucleobases. In some embodiments, provided oligonucleotides comprise at least 15 nucleobases. In some embodiments, provided oligonucleotides comprise at least 16 nucleobases. In some embodiments, provided oligonucleotides comprise at least 17 nucleobases.
• provided oligonucleotides comprise at least 18 nucleobases. In some embodiments, provided oligonucleotides comprise at least 19 nucleobases. In some embodiments, provided oligonucleotides comprise at least 20 nucleobases. In some embodiments, provided oligonucleotides comprise at least 21 nucleobases. In some embodiments, provided oligonucleotides comprise at least 22 nucleobases. In some embodiments, provided oligonucleotides comprise at least 23 nucleobases. In some embodiments, provided oligonucleotides comprise at least 24 nucleobases. In some embodiments, provided oligonucleotides comprise at least 25 nucleobases. In some embodiments, a nucleobase is optionally substituted adenine, cytosine, guanosine, thymine, or uracil, or a tautomer thereof.
• each chiral linkage phosphorus independently has a diastereomeric purity as described in the present disclosure.
• a provided compound has a purity, diastereomeric purity, and/or enantiomeric purity as described in the present disclosure.
• a provided compound has a purity as described in the present disclosure.
• a provided compound has a diastereomeric purity as described in the present disclosure.
• a provided compound has an enantiomeric purity as described in the present disclosure.
• a provided compound has a diastereomeric purity and an enantiomeric purity as described in the present disclosure.
• provided oligonucleotides comprise or are of a 5’-wing region-core region-3’-wing region structure. In some embodiments, provided oligonucleotides comprise or are of a 5’-wing region-core region structure. In some embodiments, provided oligonucleotides comprise or are of a core region-3’-wing region structure. In some embodiments, provided oligonucleotides comprise of a 5’-wing region-core region-3’-wing region structure. In some embodiments, provided oligonucleotides comprise of a 5’-wing region-core region structure. In some embodiments, provided oligonucleotides comprise of a core region-3’-wing region structure.
• provided oligonucleotides are of a 5’-wing region-core region-3’-wing region structure. In some embodiments, provided oligonucleotides are of a 5’-wing region-core region structure. In some embodiments, provided oligonucleotides are of a core region-3’-wing region structure.
• a wing-core-wing (i.e., X-Y-X) motif is represented numerically as, e.g., 5-10-4, meaning 5’-wing region is 5 bases in length, the core region is 10 bases in length, and the 3’-wing region is 4-bases in length. In some embodiments, a wing-core-wing motif is any of, e.g.
• a wing-core-wing motif is 5-10-5.
• a wing -core-wing motif is 7-7-6.
• a wing -core-wing motif is 8-7-5.
• a wing-core motif is 5-15, 6-14, 7-13, 8-12, 9-12, etc.
• a core-wing motif is 5-15, 6-14, 7-13, 8-12, 9-12, etc.
• a wing region comprises a sugar modification absent from a core region.
• a wing region comprises a 2’-modification.
• each nucleotide unit of a wing region independently comprises a 2’-modification.
• each nucleotide unit of a wing region independently comprises the same 2’-modification.
• each nucleotide unit of a 5’-wing region independently comprises the same 2’- modification.
• each nucleotide unit of a 3’-wing region independently comprises the same 2’-modification.
• 2’-modifications of the 5’-wing region are the same.
• 2’-modifications of the 5’-wing region are the different.
• a 2’- modification is 2’-OR, wherein R’ is not hydrogen.
• a 2’-modification is 2’-OR, wherein R’ is optionally substituted Ci- 6 aliphatic.
• a 2’-modification is 2’-OR, wherein R’ is optionally substituted Ci- 6 alkyl.
• a 2’-modification is 2’-OMe.
• a 2’-modification is 2’-0CH 2 CH 2 0Me.
• a wing region comprises one or more natural phosphate linkages as described in the present disclosure.
• a wing region comprises one or more non-natural intemucleotidic linkages, e.g., phosphorothioate intemucleotidic linkages.
• a core region comprises one or more natural phosphate linkages.
• a core region comprises one or more consecutive natural phosphate linkages.
• a core region comprises one or more chiral phosphate linkages.
• a core region comprises one or more consecutive chiral phosphate linkages.
• a chiral phosphate linkage is a phosphorothioate linkage.
• a chiral phosphate linkage is a chirally controlled.
• Oligonucleotides of the present disclosure may contain a pattern of backbone chiral centers.
• a pattern of backbone chiral centers of oligonucleotides or segments thereof, e.g., core regions provides increased stability.
• a pattern of backbone chiral centers provides surprisingly increased activity.
• a pattern of backbone chiral centers provides increased stability and activity.
• a pattern of backbone chiral centers provides surprisingly increased binding to certain proteins.
• a pattern of backbone chiral centers provides surprisingly enhanced delivery.
• a pattern of backbone chiral centers comprises or is (.S ' p)m(//p)n (7Zp)n(.S ' p)m. (A'p)p(//p)n(.S ' p)m. (.S ' p)p(/Zp)n(.S ' p)m. ( ⁇ Sp)m(Op)n, (Op)n( ⁇ Sp)m, (A'p)p(t/p)n(.S ' p)m.
• a pattern of backbone chiral centers comprises or is (.S ' p)m(//p)n (Ap)n(.S ' p)m (A'p)p(//p)n(.S ' p)m. (.S ' p)p(//p)n(.S ' p)m.
• a pattern of backbone chiral centers comprises or is (.S ' p)m(//p)n (Ap)n(.S ' p)m (A'p)p(//p)n(.S ' p)m.
• a pattern of backbone chiral centers comprises (.S ' p)m(//p)n (Ap)n(.S ' p)m (A'p)p(//p)n(.S ' p)m.
• a pattern of backbone chiral centers comprises ( ⁇ Sp)m(Op)n, (Op)n( ⁇ Sp)m, (A'p)p(t/p)n(.S ' p)m or (,S ' p)p(ri>p)n(,S ' p)m.
• a pattern of backbone chiral centers is (.S ' p)m(//p)n (Ap)n(.S ' p)m (A'p)p(//p)n(.S ' p)m.
• a pattern of backbone chiral centers is ( ⁇ Sp)m(Op)n, (Op)n( ⁇ Sp)m, (A'p)p(t/p)n(.S ' p)m or ( Sp ) p ( riA p ) n (Sp ) m .
• a pattern of backbone chiral centers comprises or is a repeating pattern comprising or being of (S ' p)m(//p)n. (Ap)n(.S ' p)m (A'p)p(//p)n(.S ' p)m.
• a pattern of backbone chiral centers comprises or is a repeating pattern of (.S'p)m(//p)n (Ap)n(.S'p)m (A'p)p(//p)n(.S'p)m. or (.S'p)p(//p)n(.S'p)m (from 5’ to 3’).
• a pattern of backbone chiral centers comprises or is a repeating pattern comprising or being of ( ⁇ Sp)m(Op)n, (Op)n( ⁇ Sp)m, (A'p)p(t/p)n(.S ' p)m or (,S ' p)p(ri>p)n(,S ' p)m.
• a pattern of backbone chiral centers comprises or is a repeating pattern of ( ⁇ Sp)m(Op)n, (Op)n( ⁇ Sp)m, (A'p)p(t/p)n(.S'p)m or (.Sp ) p ( riAp ) n (Sp ) m (from 5’ to 3’).
• a pattern of backbone chiral centers comprises or is (i?p)n(Sp)m, (A'p)p(//p)n(.S ' p)m. or (.S ' p)p(//p)n(.S ' p)m. wherein m > 2.
• a pattern of backbone chiral centers comprises or is (Ap)n(.S ' p)m (A'p)p(//p)n(.S ' p)m. or (.S ' p)p(//p)n(.S ' p)m. wherein n is 1, p >1, and m > 2.
• a pattern of backbone chiral centers comprises or is (Op)n( ⁇ Sp)m, (A'p)p(t/p)n(.S ' p)m or (.S ' p)p(t/p)n(.S ' p)m. wherein m > 2.
• a pattern of backbone chiral centers comprises or is (Op)n( ⁇ Sp)m, (A'p)p(t/p)n(,S ' p)m. or (.S ' p)p(Y/p)n(,S ' p)m. wherein n is 1, p >1, and m > 2.
• a pattern of backbone chiral centers of an oligonucleotide or a region thereof comprises or is two or more units independently selected from (Ap)n(,S'p)m. (A'p)p(//p)n(,S'p)m. (,S'p)p(Ap)n(,S'p)m. ( ⁇ rip)n(,S'p)m. (A'p)p( ⁇ rip)n(,S'p)m. and (.S'p)p( ⁇ rip)n(,S'p)m. wherein each variable is independently as described in the present disclosure. In some embodiments, n is 1.
• n is 1 and m of each unit is independently 2 or greater as described in the present disclosure. In some embodiments, at least two m of two units are different.
• a pattern of backbone chiral centers comprises 2, 3, 4, 5, 6, 7, 8, 9, or 10 such units. In some embodiments, a pattern of backbone chiral centers comprises 2 and no more than 2 such units. In some embodiments, a pattern of backbone chiral centers comprises 3 and no more than 3 such units. In some embodiments, a pattern of backbone chiral centers comprises 4 and no more than 4 such units. In some embodiments, a pattern of backbone chiral centers comprises 5 and no more than 5 such units.
• a region of an oligonucleotide comprises such a pattern of backbone chiral centers. In some embodiments, such a region comprises no 2’ -substitution at its sugar moieties (two 2’-H). In some embodiments, such a region is flanked by a 5’-region comprising a sugar modification as described in the present disclosure ( e.g .
• a 2’-modification e.g., 2’-OMe, 2’-MOE, 2’-F, etc., as described in the present disclosure
• a 5’-region comprising a sugar modification as described in the present disclosure (e.g., a 2’-modification, e.g., 2’-OMe, 2’-MOE, 2’-F, etc., as described in the present disclosure).
• a pattern of backbone chiral centers comprises or is (Rp)n( ⁇ Sp)m, (L ' r)r(/y)h. (A'p)p(//p)n(.S ' p)m (,S ' p)p(,S ' p)m or (.S ' p)p(//p)n(.S ' p)m
• a pattern of backbone chiral centers comprises or is (Ap)n(.S ' p)m. (,Ur)r(/y)h.
• a pattern of backbone chiral centers comprises or is (Rp)n( ⁇ Sp)m, (L ' r)r(/y)h.
• a pattern of backbone chiral centers comprises or is (Ap)n(.S ' p)m. (,Ur)r(/y)h. (A'p)p(//p)n(.S ' p)m or (.S ' p)p(/Zp)n(.S ' p)m. and the oligonucleotides comprise one or more 2’-OR modifications as described herein.
• a pattern of backbone chiral centers comprises or is (Ap)n(.S ' p)m. (,Ur)r(/y)h. (A'p)p(//p)n(.S ' p)m or (.S ' p)p(/Zp)n(.S ' p)m. and the oligonucleotides comprise one or more 2’-OR modifications, wherein R is not -H.
• a pattern of backbone chiral centers comprises or is (Rp)n( ⁇ Sp)m, (L ' r)r(/y)h.
• a pattern of backbone chiral centers comprises or is (//p)n(,S ' p)m.
• a pattern of backbone chiral centers comprises or is (Rp)n( ⁇ Sp)m. In some embodiments, a pattern of backbone chiral centers comprises or is ( ⁇ Sp)p(Rp)n. In some embodiments, a pattern of backbone chiral centers comprises or is (A'p)p(//p)n(.S ' p)m In some embodiments, a pattern of backbone chiral centers comprises (Ap)n(.S ' p)m In some embodiments, a pattern of backbone chiral centers comprises ( ⁇ Sp)p(Rp)n.
• a pattern of backbone chiral centers comprises (A'p)p(//p)n(.S ' p)m In some embodiments, a pattern of backbone chiral centers is (Ap)n(.S ' p)m In some embodiments, a pattern of backbone chiral centers is (L ' r)r(/y)h. In some embodiments, a pattern of backbone chiral centers is (A'p)p(//p)n(.S ' p)m In some embodiments, a pattern of backbone chiral centers comprises or is (,S ' p)p(,S ' p)m.
• a pattern of backbone chiral centers comprises or is (.S ' p)p(//p)n(.S ' p)m
• a pattern of backbone chiral centers comprises or is (Y/p)n(.S ' p)m. ( ⁇ Sp)p(Op)n, (A'p)p(Y/p)n(.S ' p)m. (,S ' p)p(,S ' p)m or (.S ' p)p(Y/p)n(.S ' p)m.
• a pattern of backbone chiral centers comprises or is (Y/p)n(.S ' p)m.
• a pattern of backbone chiral centers comprises or is (Y/p)n(.S ' p)m. (.S ' p)p(Y/p)n.
• a pattern of backbone chiral centers comprises or is (Op)n(Sp)m, ( ⁇ Sp)p(Op)n, (A'p)p(Y/p)n(.S ' p)m. or (.S ' p)p(Y/p)n(.S ' p)m. and the oligonucleotides comprise one or more 2’-OR modifications as described herein.
• a pattern of backbone chiral centers comprises or is (Op)n( ⁇ Sp)m, ( ⁇ Sp)p(Op)n, (A'p)p(Y/p)n(.S ' p)m. or (,S ' p)p(ri>p)n(,S ' p)m. and the oligonucleotides comprise one or more 2’-OR modifications, wherein R is not -H.
• a pattern of backbone chiral centers comprises or is (Y/p)n(.S ' p)m. (.S ' p)p(t/p)n
• a pattern of backbone chiral centers comprises or is (Y/p)n(.S ' p)m.
• (.S'p)p(t/p)n A'p(t/p)n(.S'p)m or (.S'p)p(t/p)n(.S'p)m comprises no 2’-substitutions (-CI3 ⁇ 4- at 2’-position).
• a pattern of backbone chiral centers comprises or is (Op)n( ⁇ Sp)m. In some embodiments, a pattern of backbone chiral centers comprises or is ( ⁇ Sp)p(Op)n. In some embodiments, a pattern of backbone chiral centers comprises or is (A'p)p(t/p)n(,S'p)m. In some embodiments, a pattern of backbone chiral centers comprises (Y/p)n(,S'p)m. In some embodiments, a pattern of backbone chiral centers comprises ( ⁇ Sp)p(Op)n. In some embodiments, a pattern of backbone chiral centers comprises (A'p)p(t/p)n(,S'p)m.
• a pattern of backbone chiral centers is (Y/p)n(,S ' p)m. In some embodiments, a pattern of backbone chiral centers is ( ⁇ Sp)p(Op)n. In some embodiments, a pattern of backbone chiral centers is (A'p)p(Y/p)n(,S'p)m. In some embodiments, a pattern of backbone chiral centers comprises or is (,S ' p)p(,S ' p)m.
• a pattern of backbone chiral centers comprises or is (.S'p)p(Y/p)n(.S'p)m
• an oligonucleotide, or a region thereof comprises a pattern, or a repeating pattern, of backbone chiral centers of (.S'p)m(//p)n (Ap)n(.S'p)m. (A'p)p(//p)n(.S'p)m or (.S ' p)p(/Zp)n(.S ' p)m (structurally starting from the first, and ending at the last, intemucleotidic linkage of the intemucleotidic linkages which have the pattern, or the repeating pattern, of (.S'p)m(//p)n (Ap)n(.S'p)m.
• an oligonucleotide, or a region thereof comprises a pattern, or a repeating pattern, of backbone chiral centers of ( ⁇ Sp)m(Op)n, (Op)n(Sp)m, (A'p)p(t/p)n(,S'p)m.
• a (Sp)p(i?p)n(Sp)m region ((.S'p)7(/Zp) 1 (,S'p)3) in WV-2555: mA * SmGmCmUmU * SC * ST * ST * SG * ST * SC * SC * RA * SG * SC * SmUmUmUmA * SmU) comprises no 2’-OR sugar modifications.
• each sugar moieties in the region is - ⁇ 1 ⁇ 4- at the 2’-position.
• each sugar moieties in the region is an unmodified, natural, 2’-deoxyribose moiety of DNA.
• a region comprising a pattern, or a repeating pattern, of backbone chiral centers which comprises or is (.S'p)m(//p)n (Ap)n(.S'p)m.
• (A'p)p(//p)n(.S'p)m or (.S'p)p(//p)n(.S'p)m is flanked by a 5’-wing region, which structurally ends with a nucleoside moiety (which nucleoside moiety, at its 3’-end, is connected to the first intemucleotidic linkage of the region comprising a pattern, or a repeating pattern, of backbone chiral centers which comprises or is ( ⁇ Sp)m(Rp)n, (/Zp)n(,S'p)m.
• flanking 5’-wing region in WV-2555 mA * SmGmCmUmU * SC * ST * ST * SG * ST * SC * SC * RA * SG * SC * SmUmUmA * SmU).
• a region comprising a pattern, or a repeating pattern, of backbone chiral centers which comprises or is ( ⁇ Sp)m(Op)n, (Gp)n(,Yp)m. (A'p)p(Gp)n(,Sp)m.
• (,S'p)p(Gp)n(,Yp)m is flanked by a 5’-wing region, which structurally ends with a nucleoside moiety (which nucleoside moiety, at its 3’-end, is connected to the first intemucleotidic linkage of the region comprising a pattern, or a repeating pattern, of backbone chiral centers which comprises or is ( ⁇ Sp)m(Op)n, (Gp)n(,Yp)m. (A'p)p(Gp)n(,Yp)m. or (,S'p)p(Gp)n(,Yp)m).
• a region comprising a pattern, or a repeating pattern, of backbone chiral centers which comprises or is (.Yp)m(//p)n. (Ap)n(,Yp)m. (A'p)p(//p)n(,Sp)m.
• nucleoside moiety which nucleoside moiety, at its 5’-end, is connected to the last intemucleotidic linkage of the region comprising a pattern, or a repeating pattern, of backbone chiral centers which comprises or is ( ⁇ Sp)m(Rp)n, (Ap)n(,Sp)m. (A'p)p(//p)n(,Yp)m. or (Sp)p(i?p)n(Sp)m).
• a flanking 3’-wing region in WV-2555 mA * SmGmCmUmU * SC * ST * ST * SG * ST * SC * SC * RA * SG * SC * SmUmUmUmA * SmU).
• a region comprising a pattern, or a repeating pattern, of backbone chiral centers which comprises or is ( ⁇ Sp)m(Op)n, (Gp)n(,Yp)m. (A'p)p(Gp)n(,Sp)m.
• nucleoside moiety which nucleoside moiety, at its 5’-end, is connected to the last intemucleotidic linkage of the region comprising a pattern, or a repeating pattern, of backbone chiral centers which comprises or is ( ⁇ Sp)m(Op)n, (Gp)n(,Yp)m. (A'p)p(Gp)n(,Yp)m. or (,S'p)p(Gp)n(,Yp)m).
• a region comprising a pattern, or a repeating pattern, of backbone chiral centers which comprises or is (.Yp)m(//p)n. (Ap)n(,Yp)m. (A'p)p(//p)n(,Sp)m. or (.Sp)p(//p)n(,Yp)m is flanked by a 5’- end and a 3’-wing regions.
• a region comprising a pattern, or a repeating pattern, of backbone chiral centers which comprises or is ( ⁇ Sp)m(Op)n, (Gp)n(,Yp)m. (A'p)p(Gp)n(,Yp)m.
• flanking 5’- wing region and/or the 3’-wing region comprise a non-natural intemucleotidic linkage.
• the flanking 5’-wing region and/or the 3’-wing region comprise a chiral intemucleotidic linkage.
• the flanking 5’-wing region and/or the 3’-wing region comprise a chirally controlled intemucleotidic linkage.
• flanking 5’-wing region and/or the 3’- wing region comprise a modified intemucleotidic linkage comprising a rip linkage phosphoms. In some embodiments, the flanking 5’-wing region and/or the 3’-wing region comprise a ⁇ Sp phosphorothioate linkage. In some embodiments, the flanking 5’-wing region and/or the 3’-wing region comprise one or more natural phosphate linkages. In some embodiments, the flanking 5’-wing region and/or the 3’-wing region comprise one or more consecutive natural phosphate linkages.
• the flanking 5’-end comprises only one modified intemucleotidic linkage which is the 5’-end intemucleotidic linkage, and one or more consecutive natural phosphate linkages (for example, in WV-2555: mA * SmGmCmUmU * SC * ST * ST * SG * ST * SC * SC * RA * SG * SC * SmUmUmUmA * SmU (SOOOSSSSSSSRSSSOOOS)).
• the flanking 3’-end comprises only one modified intemucleotidic linkage which is the 3’-end intemucleotidic linkage, and one or more consecutive natural phosphate linkages (for example, in WV-2555: mA * SmGmCmUmU * SC * ST * ST * SG * ST * SC * SC * RA * SG * SC * SmUmUmUmA * SmU (SOOOSSSSSSSRSSSOOOS)).
• the flanking 5’-wing region and/or the 3’-wing region comprise 2’-modified sugar units.
• each sugar unit in the 5’-wing region and/or the 3’-wing region is independently modified.
• each sugar unit in the 5’-wing region and/or the 3’-wing region independently comprises a 2’-modification (for example, m, 2’-OMe in WV-2555: mA * SmGmCmUmU * SC * ST * ST * SG * ST * SC * SC * RA * SG * SC * SmUmUmUmA * SmU).
• each sugar unit in the 5’-wing region and/or the 3’-wing region comprises the same 2’-modification.
• a 2’ -modification is 2’-OR, wherein R is optionally substituted Ci- 6 aliphatic.
• a 2’ -modification is 2’-OMe.
• a 2’ -modification is 2’-MOE.
• a 2’ -modification is a UNA modification (which comprises a type of C2-C4 bridge).
• oligonucleotides comprise 2’-F modified sugars and comprise one or more wing and one or more core (e.g., wing-core-wing, core-wing, wing-core, etc.).
• a wing e.g., a 5’-wing, a 3’-wing, etc., comprises one or more 2’-F modified sugar.
• most sugars in a wing comprises a 2’-F modification.
• each sugar in a wing comprises a 2’-F modification.
• an intemucleotidic linkages bonded to two 2’-F modified sugars is a chiral modified intemucleotidic linkage.
• each intemucleotidic linkages bonded to two 2’-F modified sugars is independently a chiral modified intemucleotidic linkage.
• each chiral modified intemucleotidic linkage is independently a phosphorothioate intemucleotidic linkage or a non-negatively charged intemucleotidic linkage.
• each chiral modified intemucleotidic linkage is independently a phosphorothioate intemucleotidic linkage or a neutral intemucleotidic linkage.
• a chiral modified intemucleotidic linkage is a phosphorothioate intemucleotidic linkage.
• each chiral modified intemucleotidic linkage is a phosphorothioate intemucleotidic linkage.
• a chiral modified intemucleotidic linkage is chirally controlled.
• a chiral modified intemucleotidic linkage is chirally controlled and is ,S ' p at the linkage phosphorus. In some embodiments, a chiral modified intemucleotidic linkage is a chirally controlled phosphorothioate intemucleotidic linkage and is Sp at the linkage phosphorus. In some embodiments, a chiral modified intemucleotidic linkage is a non-negatively charged intemucleotidic linkage and is Rp at the linkage phosphorus.
• each chirally controlled phosphorothioate intemucleotidic linkage is .
• S ' p In some embodiments, each chirally controlled phosphorothioate intemucleotidic linkage bonded to two 2’-F modified sugars is .
• S ' p In some embodiments, each intemucleotidic linkages bonded to two 2’-F modified sugars is independently a chirally controlled phosphorothioate intemucleotidic linkage and is .S ' p
• a core comprises one or more 2’-F modified sugars and one or more 2’-OR modified sugars, wherein R is as described herein.
• R is optionally substituted C M alkyl.
• R is -OMe.
• a core comprises alternating 2’-F modified sugars and 2’-OR modified sugars.
• one or more 2’-OR modified sugars are consecutive.
• the percentage of 2’-F modified sugar(s) in a core is lower that that for a wing or each wing.
• a core comprises one or more natural phosphate linkages.
• an intemucleotidic linkage bonded to two 2’-OR modified sugars is a natural phosphate linkage.
• each intemucleotidic linkage bonded to two 2’-OR modified sugars is a natural phosphate linkage.
• an intemucleotidic linkage bonded to a 2’-F modified sugar and a 2’-OR modified sugar is a natural phosphate linkage.
• each natural phosphate linkage independently bonds to a 2’ -OR modified sugar at its 3’-position.
• an intemucleotidic linkage bonded to a 2’-F modified sugar and a 2’-OR modified sugar is a modified intemucleotidic linkage (e.g., a phosphorothioate intemucleotidic linkage), which is optionally chirally controlled (and which is optionally rip at its linkage phosphoms).
• each modified intemucleotidic linkage is independently a phosphorothioate intemucleotidic linkage.
• each modified intemucleotidic linkage is chirally controlled.
• each modified intemucleotidic linkage is chirally controlled .S ' p phosphorothioate intemucleotidic linkage.
• each intemucleotidic linkage in a core is independently a natural phosphate linkage or a phosphorothioate intemucleotidic linkage.
• each intemucleotidic linkage in a core is independently a natural phosphate linkage or a chirally controlled phosphorothioate intemucleotidic linkage.
• each intemucleotidic linkage in a core is independently a natural phosphate linkage or a chirally controlled .
• S ' p intemucleotidic linkage is independently a natural phosphate linkage or a chirally controlled .
• S ' p phosphorothioate intemucleotidic linkage is independently a natural phosphate linkage or a chirally controlled .
• Example oligonucleotides comprising wing and core regions described herein include WV- 1714, WV-2444, WV-2445, WV-2526, WV-2527, WV-2528, WV-2530, WV-2531, WV-2578, WV- 2580, WV-2587, WV-3047, WV-3152, WV-3472, WV-3473, WV-3507, WV-3508, WV-3509, WV- 3510, WV-3511, WV-3512, WV-3513, WV-3514, WV-3515, WV-3545, WV-3546, WV-9517, WV- 12555, WV-12556, WV-12558, WV-12876, WV-12877, WV-12878, WV-12880, WV-13826, WV- 13835, WV-13864, or WV-14344.
• n is 1-10. In some embodiments, n is 1, 2, 3, 4, 5, 6, 7 or 8. In some embodiments, n is 1. In some embodiments, n is 2, 3, 4, 5, 6, 7 or 8. In some embodiments, n is 3, 4, 5,
• n is 4, 5, 6, 7 or 8. In some embodiments, n is 5, 6, 7 or 8. In some embodiments, n is 6, 7 or 8. In some embodiments, n is 7 or 8. In some embodiments, n is 1. In some embodiments, n is 2. In some embodiments, n is 3. In some embodiments, n is 4. In some embodiments, n is 5. In some embodiments, n is 6. In some embodiments, n is 7. In some embodiments, n is 8. In some embodiments, n is 9. In some embodiments, n is 10.
• n is 1. In some embodiments, m is at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15. In some embodiments, m is 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15. In some embodiments, p is at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15. In some embodiments, m is at least 2. In some embodiments, p is at least 2. In some embodiments, n is 1. In some embodiments, m is at least 2, p is at least 2, n is 1. In some embodiments, p is 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15. In some embodiments, each of m and p is independently at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15.
• each of m and p is independently 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15. In some embodiments, at least one of m and p is at least 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15. In some embodiments, m is at least 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15. In some embodiments, p is at least 5, 6,
• p is 0-50. In some embodiments, p is 1-50. In some embodiments, p is 1. In some embodiments, p is 2-50. In some embodiments, p is 2, 3, 4, 5, 6, 7 or 8. In some embodiments, p is 3, 4, 5, 6, 7 or 8. In some embodiments, p is 4, 5, 6, 7 or 8. In some embodiments, p is 5, 6, 7 or 8. In some embodiments, p is 6, 7 or 8. In some embodiments, p is 7 or 8. In some embodiments, p is 2. In some embodiments, p is 3. In some embodiments, p is 4. In some embodiments, p is 5. In some embodiments, p is 6. In some embodiments, p is 7. In some embodiments, p is 8.
• p is 9. In some embodiments, p is 10. In some embodiments, p is 11. In some embodiments, p is 12. In some embodiments, p is 13. In some embodiments, p is 14. In some embodiments, p is 15. In some embodiments, p is 16. In some embodiments, p is 17. In some embodiments, p is 18. In some embodiments, p is 19. In some embodiments, p is 20. In some embodiments, p is 21. In some embodiments, p is 22. In some embodiments, p is 23. In some embodiments, p is 24. In some embodiments, p is 25. In some embodiments, p is at least 2. In some embodiments, p is at least 3. In some embodiments, p is at least 4.
• p is at least 5. In some embodiments, p is at least 6. In some embodiments, p is at least 7. In some embodiments, p is at least 8. In some embodiments, p is at least 9. In some embodiments, p is at least 10. In some embodiments, p is at least 11. In some embodiments, p is at least 12. In some embodiments, p is at least 13. In some embodiments, p is at least 14. In some embodiments, p is at least 15. In some embodiments, p is at least 16. In some embodiments, p is at least 17. In some embodiments, p is at least 18. In some embodiments, p is at least 19. In some embodiments, p is at least 20. In some embodiments, p is at least 21. In some embodiments, p is at least 22. In some embodiments, p is at least 23. In some embodiments, p is at least 24. In some embodiments, p is at least 25.
• m is 0-50. In some embodiments, m is 1-50. In some embodiments, m is 1. In some embodiments, m is 2-50. In some embodiments, m is 2, 3, 4, 5, 6, 7 or 8. In some embodiments, m is 3, 4, 5, 6, 7 or 8. In some embodiments, m is 4, 5, 6, 7 or 8. In some embodiments, m is 5, 6, 7 or 8. In some embodiments, m is 6, 7 or 8. In some embodiments, m is 7 or 8. In some embodiments, m is 0. In some embodiments, m is 1. In some embodiments, m is 2. In some embodiments, m is 3. In some embodiments, m is 4. In some embodiments, m is 5. In some embodiments, m is 6.
• m is 7. In some embodiments, m is 8. In some embodiments, m is 9. In some embodiments, m is 10. In some embodiments, m is 11. In some embodiments, m is 12. In some embodiments, m is 13. In some embodiments, m is 14. In some embodiments, m is 15. In some embodiments, m is 16. In some embodiments, m is 17. In some embodiments, m is 18. In some embodiments, m is 19. In some embodiments, m is 20. In some embodiments, m is 21. In some embodiments, m is 22. In some embodiments, m is 23. In some embodiments, m is 24. In some embodiments, m is 25. In some embodiments, m is at least 2.
• m is at least 3. In some embodiments, m is at least 4. In some embodiments, m is at least 5. In some embodiments, m is at least 6. In some embodiments, m is at least 7. In some embodiments, m is at least 8. In some embodiments, m is at least 9. In some embodiments, m is at least 10. In some embodiments, m is at least 11. In some embodiments, m is at least 12. In some embodiments, m is at least 13. In some embodiments, m is at least 14. In some embodiments, m is at least 15. In some embodiments, m is at least 16. In some embodiments, m is at least 17. In some embodiments, m is at least 18. In some embodiments, m is at least 19.
• m is at least 20. In some embodiments, m is at least 21. In some embodiments, m is at least 22. In some embodiments, m is at least 23. In some embodiments, m is at least 24. In some embodiments, m is at least 25. In some embodiments, m is at least greater than 25.
• At least one of m and p is greater than 2. In some embodiments, at least one of m and p is greater than 3. In some embodiments, at least one of m and p is greater than 4. In some embodiments, at least one of m and p is greater than 5. In some embodiments, at least one of m and p is greater than 6. In some embodiments, at least one of m and p is greater than 7. In some embodiments, at least one of m and p is greater than 8. In some embodiments, at least one of m and p is greater than 9. In some embodiments, at least one of m and p is greater than 10. In some embodiments, at least one of m and p is greater than 11.
• At least one of m and p is greater than 12. In some embodiments, at least one of m and p is greater than 13. In some embodiments, at least one of m and p is greater than 14. In some embodiments, at least one of m and p is greater than 15. In some embodiments, at least one of m and p is greater than 16. In some embodiments, at least one of m and p is greater than 17. In some embodiments, at least one of m and p is greater than 18. In some embodiments, at least one of m and p is greater than 19. In some embodiments, at least one of m and p is greater than 20. In some embodiments, at least one of m and p is greater than 21.
• At least one of m and p is greater than 22. In some embodiments, at least one of m and p is greater than 23. In some embodiments, at least one of m and p is greater than 24. In some embodiments, at least one of m and p is greater than 25.
• each of m and p is greater than 2. In some embodiments, each of m and p is greater than 3. In some embodiments, each of m and p is greater than 4. In some embodiments, each of m and p is greater than 5. In some embodiments, each of m and p is greater than 6. In some embodiments, each of m and p is greater than 7. In some embodiments, each of m and p is greater than 8. In some embodiments, each of m and p is greater than 9. In some embodiments, each of m and p is greater than 10. In some embodiments, each of m and p is greater than 11. In some embodiments, each of m and p is greater than 12.
• each of m and p is greater than 13. In some embodiments, each of m and p is greater than 14. In some embodiments, each of m and p is greater than 15. In some embodiments, each of m and p is greater than 16. In some embodiments, each of m and p is greater than 17. In some embodiments, each of m and p is greater than 18. In some embodiments, each of m and p is greater than 19. In some embodiments, each of m and p is greater than 20.
• the sum of m and p is greater than 3. In some embodiments, the sum of m and p is greater than 4. In some embodiments, the sum of m and p is greater than 5. In some embodiments, the sum of m and p is greater than 6. In some embodiments, the sum of m and p is greater than 7. In some embodiments, the sum of m and p is greater than 8. In some embodiments, the sum of m and p is greater than 9. In some embodiments, the sum of m and p is greater than 10. In some embodiments, the sum of m and p is greater than 11. In some embodiments, the sum of m and p is greater than 12. In some embodiments, the sum of m and p is greater than 13.
• the sum of m and p is greater than 14. In some embodiments, the sum of m and p is greater than 15. In some embodiments, the sum of m and p is greater than 16. In some embodiments, the sum of m and p is greater than 17. In some embodiments, the sum of m and p is greater than 18. In some embodiments, the sum of m and p is greater than 19. In some embodiments, the sum of m and p is greater than 20. In some embodiments, the sum of m and p is greater than 21.
• the sum of m and p is greater than 22 In some embodiments, the sum of m and p is greater than 23 In some embodiments, the sum of m and p is greater than 24 In some embodiments, the sum of m and p is greater than 25
• n is 1, and at least one of m and p is greater than 1. In some embodiments, n is i and each of m and p is independently greater than 1. In some embodiments, m>n and p>n. In some embodiments, (.S ' p)m(/Zp)n(.S ' p)p is (.S ' pb/ZpOS ' pb. In some embodiments,
• (.S ' p)p(/Zp)n(.S ' p)m is ( r) 2 / ⁇ r( r)2. In some embodiments, (.S ' p)p(/Zp)n(.S ' p)m is r/ ⁇ r( r) 2 . In some embodiments, (A'p)p(//p)n(.S ' p)m is (A'p)t//p(.S ' p)m In some embodiments, (A'p)p(//p)n(.S ' p)m is (A'pb/ZpOS ' pIm.
• (A'p)p(//p)n(.S ' p)m is (/y) 2 /y(L ' r)h ⁇ . In some embodiments, (A'p)p(//p)n(.S ' p)m is (.Sp) 2 / ⁇ p(,Sp)m . In some embodiments, (A'p)p(//p)n(.S ' p)m is Ap.S ' p//p(.S ' p)m In some embodiments, (A'p)p(//p)n(.S ' p)m is .S ' p//p//p(.S ' p)m
• (.S ' p)p(//p)n(.S ' p)m is L ' r/y,Ur,Ur. In some embodiments,
• ( ⁇ Yp)p(Rp)n(Yp)m is (.Sp) 2 / ⁇ p(,Sp) 2 .
• (.Yp)p(//p)n(,Yp)m is (.Vp) /p(.Vp) 2
• (.Yp)p(//p)n(,Yp)m is ( ⁇ Ur) /f(Ur) 4 .
• (.Yp)p(//p)n(,Yp)m is (.Yp)t//p(.Yp) ⁇ .
• (.S ' p)p(//p)n(.S ' p)m is .Yp//p(,YpE. In some embodiments, (.S ' p)p(//p)n(.S ' p)m is (.S ' p) 2 / ⁇ p(.S ' p) ⁇ . In some embodiments, (.Yp)p(//p)n(,Yp)m is (,Ur) 4 / (,Ur)5. In some embodiments, (.Yp)p(//p)n(,Yp)m is (.S ' p) / ⁇ p(.S ' p) ⁇ . In some embodiments, (.Yp)p(//p)n(,Yp)m is (.Yp)J/p(,Yp) ⁇ .
• ( ⁇ Yp)m(Rp)n(Yp)p is (.Vp) 2 / ⁇ p(.Vp) 2 . In some embodiments,
• (.Yp)m(//p)n(,Yp)p is (.Sp) 2 / ⁇ p(,Sp) 2 .
• (.Yp)m(//p)n(,Yp)p is (.S ' p) 4 / ⁇ p(.S ' p)4.
• ( ⁇ Yp)m(Rp)n(Yp)p is (.Ur)hi/y(,UrE.
• (.Yp)m(//p)n(,Yp)p is (.S ' p) 2 / ⁇ p(.S ' p) ⁇ .
• (.Yp)m(//p)n(,Yp)p is (.S ' p) 2 / ⁇ p(.S ' p) ⁇ .
• (.Yp)m(//p)n(,Yp)p is (.S ' p) 4 //p(.S ' p) ⁇ .
• (.Yp)m(//p)n(,Yp)p is (.Yp)J/p(,Yp) ⁇ .
• provided oligonucleotides are blockmers. In some embodiments, provided oligonucleotide are altmers. In some embodiments, provided oligonucleotides are altmers comprising alternating blocks.
• a blockmer or an altmer can be defined by chemical modifications (including presence or absence), e.g. , base modifications, sugar modification, intemucleotidic linkage modifications, stereochemistry, etc., or patterns thereof. Example chemical modifications, stereochemistry and patterns thereof for a block and/or an alternating unit include but are not limited to those described in this disclosure, such as those described for an oligonucleotide, etc.
• a blockmer comprises a pattern of ..SS..RR..SS..RR...
• an altmer comprises a pattern of SRSRSRSR.
• a provided pattern of backbone chiral centers comprises repeating (.S ' p)m(//p)n (//p)n(,S ' p)m. (A'p)p(//p)n(.S ' p)m or (.Yp)p(//p)n(,Yp)m units.
• a repeating unit is (.S ' p)m(//p)n
• a repeating unit is L ' r/y.
• a repeating unit is SpSpRp.
• a repeating unit is SpRpRp.
• a repeating unit is RpRpSp. In some embodiments, a repeating unit is (/Zp)n(,S'p)m. In some embodiments, a repeating unit is (A'p)p(//p)n(,S'p)m. In some embodiments, a repeating unit is (.S'p)p(/Zp)n(,S'p)m.
• oligonucleotides of the present disclosure comprise base sequences, base modifications, sugar modifications, pattern of backbone linkages (intemucleotidic linkages), and/or pattern of backbone chiral centers (e.g., of linkage phosphorus atoms) as described in US20150211006, US20170037399, US 20180216107, US 20180216108, US 20190008986, WO 2017/015555, WO
• provided technologies comprise labeling of oligonucleotides, e.g., using isotopes.
• provided oligonucleotides contain increased levels of one or more isotopes.
• provided oligonucleotides are labeled, e.g., by one or more isotopes of one or more elements, e.g., hydrogen, carbon, nitrogen, etc.
• provided oligonucleotides in provided compositions comprise one or more base modifications, sugar modifications, and/or intemucleotidic linkage modifications, wherein the oligonucleotides contain an enriched level of an isotope.
• an isotope is deuterium.
• a hydrogen in a sugar is replaced by deuterium (e.g., at the 2’ position of a 2’- deoxy).
• a hydrogen in a base is replaced by deuterium.
• a hydrogen in an intemucleotidic linkage is replaced by deuterium.
• provided oligonucleotides are labeled with deuterium (replacing -3 ⁇ 4 with - 2 H) at one or more positions.
• replacement of a hydrogen with deuterium can improve the stability, activity, bioavailability, easy of use, convenience, efficacy, and/or systemic exposure of an oligonucleotide.
• one or more 'H of an oligonucleotide or any moiety conjugated to the oligonucleotide e.g., a targeting moiety, lipid, etc.
• Such oligonucleotides can be used in any composition or method described herein.
• an oligonucleotide which targets HTT comprises one or more isotopes.
• an oligonucleotide which targets dystrophin comprises one or more isotopes.
• provided technologies are particularly useful for preparing chirally controlled oligonucleotide composition with high (crude) purity and/or yield.
• chiral auxiliaries are typically used to control stereochemistry of a formed linkage phosphorus chiral center.
• the present disclosure provides compounds, e.g., of formula I, I-a, I-a-1, 1-a-2, 1-b, I-c, I- d, I-e, II, Il-a, Il-b, III, Ill-a, or Ill-b, or salts thereof, that may be utilized as chiral auxiliaries for oligonucleotide synthesis.
• a chiral auxiliary is one described in US 9598458, US 9744183, US 9605019, US 9394333, US 9598458, US 8859755, US 20130178612, US 8470987, US 8822671, US 20150211006, US 20170037399, US 20180216107, US 20180216108, US 20190008986, WO 2017/015555, WO 2017/015575, WO 2017/062862, WO 2017/160741, WO 2017/192664, WO 2017/192679, WO 2017/210647, WO 2018/022473, WO 2018/067973, WO 2018/098264, WO
• the present disclosure provides a compound having the structure of formula I:
• L is a covalent bond, or optionally substituted Ci- 6 alkylene, wherein one or more methylene units are optionally and independently replaced with -L’-; or L is L s ;
• each L’ is independently a covalent bond, optionally substituted bivalent C1-3 alkylene,
• each of R 1 , R 2 , R 3 , R 4 , and R 5 is independently -H, -L s -R’, halogen, -CN, -N0 2 , -L s -Si(R’)3,
• each L s is independently a covalent bond, or a bivalent, optionally substituted, linear or branched group selected from a Ci-30 aliphatic group and a Ci-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 group selected from Ci- 6 alkylene, Ci- 6 alkenylene, CoC .
• each -Cy- is independently an optionally substituted bivalent group selected from a C3-20 cycloaliphatic ring, a CV20 aryl ring, a 5-20 membered heteroaryl ring having 1-10 heteroatoms independently selected from oxygen, nitrogen, sulfur, phosphorus and silicon, and a 3-20 membered heterocyclyl ring having 1-10 heteroatoms independently selected from oxygen, nitrogen, sulfur, phosphorus and silicon;
• each Cy L is independently an optionally substituted tetravalent group selected from a C3-20 cycloaliphatic ring, a CV20 aryl ring, a 5-20 membered heteroaryl ring having 1-10 heteroatoms independently selected from oxygen, nitrogen, sulfur, phosphorus and silicon, and a 3-20 membered heterocyclyl ring having 1-10 heteroatoms independently selected from oxygen, nitrogen, sulfur, phosphorus and silicon;
• each R’ is independently -R, -C(0)R, -CO2R, or -SO2R;
• R 6 is -L-R’ ;
• R 7 is -OH or -SH
• R 1 , R 2 , R 3 and R 4 is not -H;
• each R is independently -H, or an optionally substituted group selected from Ci-30 aliphatic, Ci-30 heteroaliphatic having 1-10 heteroatoms independently selected from oxygen, nitrogen, sulfur, phosphorus and silicon, Ce-30 aryl, Ce-30 arylaliphatic, Ce-30 arylheteroaliphatic having 1-10 heteroatoms independently selected from oxygen, nitrogen, sulfur, phosphorus and silicon, 5-30 membered heteroaryl having 1-10 heteroatoms independently selected from oxygen, nitrogen, sulfur, phosphorus and silicon, and 3-30 membered heterocyclyl having 1-10 heteroatoms independently selected from oxygen, nitrogen, sulfur, phosphorus and silicon, or
• R groups on the same atom are optionally and independently taken together with the atom to form an optionally substituted, 3-30 membered, monocyclic, bicyclic or polycyclic ring having, in addition to the atom, 0-10 heteroatoms independently selected from oxygen, nitrogen, sulfur, phosphorus and silicon; or
• L is a covalent bond, or optionally substituted Ci- 6 alkylene, wherein one or more methylene units are optionally and independently replaced with -L’-.
• L is a covalent bond.
• a provided compound has the structure of
• R R or a salt thereof are taken together with their intervening atoms to form an optionally substituted, 3-20 membered monocyclic, bicyclic or polycyclic ring having 1-5 heteroatoms.
• one of R 1 and R 2 are taken together with R 5 and their intervening atoms to form an optionally substituted, 3-20 membered monocyclic, bicyclic or polycyclic ring having 1-5 heteroatoms.
• a formed ring can be of various sizes, monocyclic, bicyclic or polycyclic, and contain various numbers of heteroatoms.
• a ring is a 3-membered ring.
• a ring is a 4-membered ring. In some embodiments, a ring is a 5 -membered ring. In some embodiments, a ring is a 6-membered ring. In some embodiments, a ring is monocyclic. In some embodiments, a ring contains additional ring heteroatoms other than the intervening heteroatoms. In some embodiments, a ring is a 3-membered ring containing one ring heteroatom. In some embodiments, a ring is a 3-membered ring containing two ring heteroatoms. In some embodiments, a ring is a 3-membered ring containing one carbon, one nitrogen, and one oxygen ring atom.
• L is -C(R 3 )(R 4 )-.
• a provided compound has the structure of formula I-a:
• a compound of formula I has the structure of formula I-a. In some embodiments, a
• provided compound has the structure salt thereof, wherein each variable is independently as described in the present disclosure, wherein R 4 and R 5 are not hydrogen.
• a provided compound has the structure of formula (I-a-1):
• a compound of formula I-a has the structure of formula I-a-1.
• a provided compound has the structure of formula (I-a-2):
• a compound of formula I-a has the structure of formula I-a-2.
• R 6 is -H. In some embodiments, R 6 is -H, and R 4 and R 5 are taken together with their intervening atoms to form an optionally substituted 3-20 membered heterocyclyl ring having 1-5 heteroatoms. In some embodiments, R 4 and R 5 are taken together with their intervening atoms to form an optionally substituted 4-10 membered heterocyclyl ring with the intervening nitrogen atom as the only ring heteroatom. In some embodiments, a formed ring is 3-membered. In some embodiments, a formed ring is 4-membered. In some embodiments, a formed ring is 5-membered. In some embodiments, a formed ring is 6-membered.
• a formed ring is 7-membered. In some embodiments, a formed ring is substituted. In some embodiments, a formed ring is unsubstituted. In some embodiments, a formed ring is monocyclic. In some embodiments, a formed ring is bicyclic. In some embodiments, a formed ring is polycyclic. In some embodiments, a formed ring is saturated. In some embodiments, a formed ring has no ring heteroatoms in addition to the nitrogen to which R 5 is attached.
• R 1 and R 2 are not hydrogen. In some embodiments, R 1 is hydrogen and R 2 is not hydrogen. In some embodiments, R 1 is not hydrogen and R 2 is hydrogen. In some embodiments, neither of R 1 and R 2 is hydrogen.
• one of R 1 and R 2 is -H, and the other is R, wherein R is not hydrogen. In some embodiments, one of R 1 and R 2 is -H, and the other is optionally substituted Ci- 6 aliphatic. In some embodiments, one of R 1 and R 2 is -H, and the other is optionally substituted CM aliphatic. In some embodiments, one of R 1 and R 2 is -H, and the other is optionally substituted C1-3 aliphatic. In some embodiments, one of R 1 and R 2 is -H, and the other is optionally substituted C1-2 aliphatic.
• one of R 1 and R 2 is -H, and the other is optionally substituted C M alkenyl. In some embodiments, one of R 1 and R 2 is -H, and the other is vinyl. In some embodiments, one of R 1 and R 2 is -H, and the other is optionally substituted CM alkynyl. In some embodiments, one of R 1 and R 2 is -H, and the other is ethynyl. In some embodiments, one of R 1 and R 2 is -H, and the other is optionally substituted benzyl. In some embodiments, one of R 1 and R 2 is -H, and the other is benzyl wherein the phenyl group of the benzyl is optionally substituted.
• R 1 is -H and R 2 is benzyl. In some embodiments, R 1 is -H and R 2 is -R, wherein R is as described in the present disclosure and is not hydrogen. In some embodiments, R 2 is optionally substituted CM aliphatic. In some embodiments, R 2 is optionally substituted -Cff-CPI ⁇ Me. In some embodiments, R 2 is -Cff-CPI ⁇ Me. In some embodiments, R 2 is optionally substituted phenyl. In some embodiments, R 2 is optionally substituted
• a provided compound is thereof.
• R 1 is not -H and R 2 is not -H. In some embodiments, R 1 and R 2 are independently R, wherein R is not -H. In some embodiments, R 1 is optionally substituted Ci- 6 aliphatic, and R 2 is optionally substituted phenyl. In some embodiments, R 1 is methyl and R 2 is phenyl.
• R 1 and R 2 are R, wherein R comprises a ring moiety.
• R is an optionally substituted group selected from C 3-20 cycloaliphatic, CV20 aryl, 5-20 membered heteroaryl having 1-5 heteroatoms, and 3-20 membered heterocyclyl having 1-5 heteroatoms.
• R is optionally substituted C 3-20 cycloaliphatic.
• R is optionally substituted C 3-10 cycloaliphatic.
• R is optionally substituted C 3-10 cycloalkyl.
• R is optionally substituted C 4-10 cycloalkyl.
• R is optionally substituted cyclopropyl. In some embodiments, R is optionally substituted cyclobutyl. In some embodiments, R is optionally substituted cyclopentyl. In some embodiments, R is optionally substituted cyclohexyl. In some embodiments, R is optionally substituted cycloheptyl. In some embodiments, R is cyclopropyl. In some embodiments, R is cyclobutyl. In some embodiments, R is cyclopentyl. In some embodiments, R is cyclohexyl. In some embodiments, R is cycloheptyl. In some embodiments, R is optionally substituted CV20 aryl.
• R is optionally substituted phenyl. In some embodiments, R is phenyl. In some embodiments, R is optionally substituted 5-20 membered heteroaryl having 1-5 heteroatoms. In some embodiments, R is optionally substituted 5- membered heteroaryl having 1-5 heteroatoms. In some embodiments, R is optionally substituted 6- membered heteroaryl having 1-5 heteroatoms. In some embodiments, R is optionally substituted 3-20 membered heterocyclyl having 1-5 heteroatoms. In some embodiments, the other of R 1 and R 2 is R wherein R is not hydrogen. In some embodiments, R is optionally substituted Ci- 6 aliphatic. In some embodiments, R is optionally substituted Ci- 6 alkyl.
• R is Ci- 6 alkyl. In some embodiments, R is methyl. In some embodiments, R is substituted methyl. In some embodiments, R is ethyl. In some embodiments, R is substituted ethyl. In some embodiments, one of R 1 and R 2 is R comprising a cyclic moiety as described in the present disclosure, and the other is an alkyl group as described in the present disclosure.
• each of R 1 and R 2 is independently R, wherein R is optionally substituted Ci-20 aliphatic. In some embodiments, R is unsubstituted Ci-20 aliphatic. In some embodiments, R is optionally substituted Ci-20 alkyl. In some embodiments, R is optionally substituted Ci- 6 alkyl. In some embodiments, R is linear Ci- 6 alkyl. In some embodiments, one of R 1 and R 2 is optionally substituted Ci- 6 alkyl, and the other is optionally substituted Ci- 6 alkyl. In some embodiments, R 1 and R 2 are the same. In some embodiments, R 1 and R 2 are different.
• one of R 1 and R 2 is optionally substituted Ci- 6 alkyl, and the other is optionally substituted Ci- 6 alkenyl. In some embodiments, one of R 1 and R 2 is optionally substituted methyl or ethyl, and the other is vinyl. In some embodiments, one of R 1 and R 2 is methyl, and the other is vinyl.
• one of R 1 and R 2 is optionally substituted Ci- 6 alkyl, and the other is optionally substituted Ci- 6 alkynyl. In some embodiments, one of R 1 and R 2 is optionally substituted methyl or ethyl, and the other is ethynyl. In some embodiments, one of R 1 and R 2 is methyl, and the other is ethynyl.
• one of R 1 and R 2 is optionally substituted Ci- 6 alkyl, and the other is optionally substituted Ci- 6 alkyl.
• R 1 and R 2 are the same optionally substituted Ci- 6 alkyl.
• R 1 and R 2 are the same optionally substituted C1-2 alkyl, and R 1 and R 2 comprise no more than two carbon atoms.
• both R 1 and R 1 are methyl.
• both R 1 and R 1 are ethyl.
• both R 1 and R 1 are isopropyl.
• one of R 1 and R 2 is optionally substituted C1-3 linear alkyl, and the other is optionally substituted C3-10 cycloalkyl. In some embodiments, one of R 1 and R 2 is optionally substituted C1-3 linear alkyl, and the other is optionally substituted C5-6 cycloalkyl. In some embodiments, R 1 is methyl. In some embodiments, R 2 is cyclopentyl. In some embodiments, R 2 is cyclohexyl. In some embodiments, one of R 1 and R 2 is optionally substituted C1-3 linear alkyl, and the other is optionally substituted benzyl. In some embodiments, R 1 is methyl and R 2 is optionally substituted benzyl.
• a provided compound is or a salt thereof. In some embodiments, a provided compound is or a salt thereof. In some embodiments, a provided compound is or a salt thereof. In some embodiments, a
• provided compound is or a salt thereof. In some embodiments, a provided compound is or a salt thereof. In some embodiments, a provided compound is or a salt thereof. In some embodiments, a provided compound
• a provided compound i salt thereof. In some embodiments, a provided compound is or a sa it thereof. In some
• a provided compound is MePh 2 Si ⁇ '" ‘‘O or a salt thereof. In some embodiments, a provided compound is or a salt thereof.
• one of R 1 and R 2 is optionally substituted Ci- 6 alkyl, and the other is optionally substituted phenyl.
• R 1 is methyl
• R 2 is optionally substituted phenyl.
• R 1 is methyl
• R 2 is phenyl.
• R 1 is methyl
• i n some embodiments, a
• provided compound is selected from ,
• a provided compound is F or
• a salt thereof in some embodiments, a provided compound salt thereof.
• a provided compound salt thereof a provided compound salt thereof.
• R 1 and R 2 are independently R, wherein R is an optionally substituted aryl group. In some embodiments, R 1 and R 2 are independently optionally substituted phenyl. In some
• R 1 and R 2 are phenyl
• a provided compound is or a salt thereof.
• R 1 and R 2 are taken together with the carbon atom they are attached on to form an optionally substituted 3-20 membered monocyclic, bicyclic or polycyclic ring having 0-5 heteroatoms. In some embodiments, R 1 and R 2 are taken together with the carbon atom they are attached on to form an optionally substituted 3-7 membered monocyclic ring having no heteroatoms.
• such a formed monocyclic ring is 3 -membered; in some embodiments, 4-membered; in some embodiments, 5 -membered; in some embodiments, 6-membered; in some embodiments 7- membered; in some embodiments, 8-membered; in some embodiments 9-membered; and in some embodiments 10-membered.
• a formed ring is monocyclic.
• a formed ring is bicyclic.
• a formed ring is polycyclic.
• a formed ring is aliphatic.
• a formed ring comprises no unsaturation.
• a formed ring is saturated, partially unsaturated, and/or partially aromatic, for example, a bicyclic or polycyclic ring comprising fused saturated, partially unsaturated, and/or aromatic moieties.
• such a formed ring is substituted.
• such a formed ring is not substituted.
• the carbon to which R 1 and R 2 are attached is not chiral.
• R 1 and R 2 are the same, and the carbon they are attached on is not chiral.
• the ring formed by R 1 and R 2 taken together with the carbon atom they are attached on does not introduce asymmetry, and the carbon atom R 1 and R 2 attached on is not chiral.
• R 1 and R 2 are different, and the carbon they are attached on is chiral. In some embodiments, the ring formed by R 1 and R 2 taken together with the carbon atom they are attached on introduces asymmetry, and the carbon atom R 1 and R 2 attached on is not chiral. In some embodiments, a provided compound is
• a provided compound is selected from salts thereof. In some embodiments, a provided compound
• a provided compound salt thereof thereof.
• a provided compound i salt thereof in some embodiments, a provided compound i salt thereof.
• R 4 and R 5 are taken together with their intervening atoms to form an optionally substituted 3-20 membered monocyclic, bicyclic or polycyclic ring having 1-5 heteroatoms.
• a formed ring is 3-membered.
• a formed ring is 4-membered.
• a formed ring is 5-membered.
• a formed ring is 6- membered.
• a formed ring is 7-membered.
• a formed ring is 8-membered.
• a formed ring is 9-membered.
• a formed ring is 10-membered.
• R 3 is -H, and R 4 and R 5 are taken together with their intervening atoms to form an optionally substituted 3-20 membered monocyclic, bicyclic or polycyclic ring having 1- 5 heteroatoms. In some embodiments, R 3 is -H, and R 4 and R 5 are taken together with their intervening atoms to form an optionally substituted 3-20 membered monocyclic, bicyclic or polycyclic ring having a nitrogen atom (the one which R 5 is on). In some embodiments, R 3 is -H, and R 4 and R 5 are taken together with their intervening atoms to form an optionally substituted 4-7 membered monocyclic ring having a nitrogen atom (the one which R 5 is on).
• R 3 is -H, and R 4 and R 5 are taken together with their intervening atoms to form an optionally substituted 4-membered monocyclic ring having a nitrogen atom (the one which R 5 is on). In some embodiments, R 3 is -H, and R 4 and R 5 are taken together with their intervening atoms to form an optionally substituted 5-membered monocyclic ring having a nitrogen atom (the one which R 5 is on). In some embodiments, R 3 is -H, and R 4 and R 5 are taken together with their intervening atoms to form an optionally substituted 6-membered monocyclic ring having a nitrogen atom (the one which R 5 is on).
• R 3 is -H, and R 4 and R 5 are taken together with their intervening atoms to form an optionally substituted 7-membered monocyclic ring having a nitrogen atom (the one which R 5 is on). In some embodiments, R 3 is -H, and R 4 and R 5 are taken together with their intervening atoms to form an optionally substituted 8-membered monocyclic ring having a nitrogen atom (the one which R 5 is on). In some embodiments, R 3 is -H, and R 4 and R 5 are taken together with their intervening atoms to form an optionally substituted 9-membered monocyclic ring having a nitrogen atom (the one which R 5 is on).
• R 3 is -H
• R 4 and R 5 are taken together with their intervening atoms to form an optionally substituted 10-membered monocyclic ring having a nitrogen atom (the one which R 5 is on).
• a ring formed by R 4 and R 5 taken together with their intervening atoms is substituted.
• a ring formed by R 4 and R 5 taken together with their intervening atoms is unsubstituted.
• a ring formed by R 4 and R 5 taken together with their intervening atoms is monocyclic.
• a ring formed by R 4 and R 5 taken together with their intervening atoms is bicyclic.
• one of R 1 and R 2 , and one of R 3 and R 4 are taken together with their intervening atoms to form an optionally substituted 3-20 membered monocyclic, bicyclic or polycyclic ring having 0-5 heteroatoms.
• a formed ring is a 3 -membered ring.
• a formed ring is a 4-membered ring.
• a formed ring is a 5 -membered ring.
• a formed ring is a 6-membered ring.
• a formed ring is a 7-membered ring.
• a formed ring is substituted.
• a formed ring is unsubstituted. In some embodiments, a formed ring is monocyclic. In some embodiments, a formed ring is bicyclic. In some embodiments, a formed ring is polycyclic. In some embodiments, a formed ring has no additional heteroatoms in addition to an intervening atom. In some embodiments, a formed ring has additional ring heteroatoms in addition to an intervening atom. Example rings formed are extensively described in the present disclosure. In
• a provided compound is selected from
• a provided compound is selected from salts
• a provided compound salt thereof thereof.
• a provided compound i salt thereof in some embodiments, a provided compound i salt thereof.
• a provided compound is or a salt thereof. In some embodiments, a provided compound is
• a provided compound a salt thereof.
• a provided compound is or a salt thereof.
• a provided compound or a salt thereof in some embodiments, a provided compound or a salt thereof.
• a provided compound is
• one or two of R 1 and R 2 are taken together with one or more of R 3 , R 4 , and R 5 and the intervening atoms to form an optionally substituted 3-20 membered monocyclic, bicyclic or polycyclic ring having 0-5 heteroatoms.
• one or two of R 1 and R 2 are taken together with one or two of R 3 and R 4 and the intervening atoms to form an optionally substituted 3-20 membered monocyclic, bicyclic or polycyclic ring having 0-5 heteroatoms.
• one or two of R 1 and R 2 are taken together with R 5 and the intervening atoms to form an optionally substituted 3- 20 membered monocyclic, bicyclic or polycyclic ring having 1-5 heteroatoms. In some embodiments, one or two of R 1 and R 2 are taken together with R 5 , one or two of R 3 and R 4 , and the intervening atoms to form an optionally substituted 3-20 membered monocyclic, bicyclic or polycyclic ring having 1-5 heteroatoms.
• one or two of R 1 and R 2 are taken together with R 5 , one or two of R 3 and R 4 , and the intervening atoms to form an optionally substituted 6-20 membered bicyclic or polycyclic ring having 1-5 heteroatoms.
• one or two of R 1 and R 2 are taken together with R 5 , one or two of R 3 and R 4 , and the intervening atoms to form an optionally substituted 8-20 membered bicyclic or polycyclic ring having 1-5 heteroatoms.
• one of R 1 and R 2 are taken together with R 5 , one of R 3 and R 4 , and the intervening atoms to form an optionally substituted 8-20 membered bicyclic or polycyclic ring having 1-5 heteroatoms. In some embodiments, one of R 1 and R 2 are taken together with R 5 , one of R 3 and R 4 , and the intervening atoms to form an optionally substituted 8-20 membered bicyclic ring having 1-5 heteroatoms. In some embodiments, a formed ring is 8- membered. In some embodiments, a formed ring is 9-membered.
• R 5 is taken with one of R 1 and R 2 and their intervening atoms to form an optionally substituted 3-20 membered monocyclic, bicyclic or polycyclic ring having 1-5 heteroatoms. In some embodiments, R 5 is taken with one of R 3 and R 4 and their intervening atoms to form an optionally substituted 3-20 membered monocyclic, bicyclic or polycyclic ring having 1-5 heteroatoms.
• Example rings formed are extensively described in the present disclosure. In some embodiments, a formed ring is 3-membered. In some embodiments, a formed ring is 4-membered. In some embodiments, a formed ring is 5-membered. In some embodiments, a formed ring is 6-membered.
• R 5 is not taken with R 1 , R 2 , R 3 , or R 4 to form an optionally substituted ring.
• R 5 is optionally substituted Ci- 6 aliphatic.
• R 5 is optionally substituted Ci- 6 alkyl.
• R 5 is unsubstituted Ci- 6 alkyl.
• R 5 is methyl.
• R 5 is ethyl.
• R 5 is isopropyl.
• a provided compound salt thereof in some embodiments, a provided compound salt thereof.
• a provided compound salt thereof In some embodiments, a provided compound salt thereof.
• L is -L’-C(R 3 )(R 4 )-.
• a provided compound has the structure of formula I-b:
• a compound of formula I has the structure of formula I-b.
• L’ is a covalent bond. In some embodiments, L’ is -C(R 3 )(R 4 )-. In some embodiments, a provided compound has the structure of formula I-c:
• a compound of formula I has the structure of formula I-c.
• one or R 3 and R 4 on C2 are taken together with R 5 to form with their intervening atoms an optionally substituted 3-20 membered monocyclic, bicyclic or polycyclic ring having a nitrogen atom (the one which R 5 is on).
• one or R 3 and R 4 on C3 are taken together with R 5 to form with their intervening atoms an optionally substituted 3-20 membered monocyclic, bicyclic or polycyclic ring having a nitrogen atom (the one which R 5 is on).
• one of R 3 and R 4 on C2, and one of R 3 and R 4 on C3, are taken together with their intervening atoms to form an optionally substituted 3-20 membered monocyclic, bicyclic or polycyclic ring having 0-5 heteroatoms.
• R 3 and R 4 on the same carbon atom are taken together with the carbon atom to form an optionally substituted 3-20 membered monocyclic, bicyclic or polycyclic ring having 0-5 heteroatoms.
• R 3 and R 4 on C2 are taken together with C2 to form an optionally substituted 3-20 membered monocyclic, bicyclic or polycyclic ring having 0-5 heteroatoms.
• R 3 and R 4 on C3 are taken together with C3 to form an optionally substituted 3-20 membered monocyclic, bicyclic or polycyclic ring having 0-5 heteroatoms.
• Example such ring moieties e.g., formed by R 3 /R 4 and R 5 , by R 3 /R 4 and R 3 /R 4 , etc., are extensively described in the present disclosure, and can be e.g., 4-membered, 5-membered, 6-membered, 7-membered, monocyclic, bicyclic, polycyclic, substituted, unsubstituted, with additional ring heteroatoms (other than the intervening atom(s)), without additional ring hetereoatoms, combinations thereof, etc.
• R 3 on C2 is hydrogen. In some embodiments, R 4 on C2 is hydrogen. In some embodiments, R 3 on C3 is hydrogen. In some embodiments, R 4 on C3 is hydrogen. In some embodiments, both R 3 and R 4 on C2 are hydrogen. In some embodiments, both R 3 and R 4 on C3 are hydrogen. In some embodiments, both R 3 and R 4 on C2, and one of R 3 and R 4 on C3, are hydrogen. In some embodiments, both R 3 and R 4 on C3, and one of R 3 and R 4 on C2, are hydrogen.
• a provided compound or a salt thereof in some embodiments, a provided compound or a salt thereof.
• L is -Cy-.
• a provided compound has the structure of formula I-d:
• a compound of formula I has the structure of formula I-d.
• -Cy- is 1,2-bivalent.
• -Cy- is optionally substituted cycloalkylene.
• -Cy- is optionally substituted .
• -Cy- is optionally
• one of R 1 and R 2 , and one of R 3 and R 4 are R and are taken together with their intervening atoms to form an optionally substituted 3-20 membered ring having 1-10 heteroatoms as described in the present disclosure, e.g., Ring A as described herein.
• a provided compound has the structure of formula I-e:
• a compound of formula I has the structure of formula I-e.
• one of R 1 and R 2 , and R 4 are taken together with their intervening atoms to form an optionally substituted 3-20 membered ring having 1-5 heteroatoms.
• R 3 is -H
• one of R 1 and R 2 , and R 4 are taken together with their intervening atoms to form an optionally substituted 3-20 membered ring having 1-5 heteroatoms.
• R 2 and R 4 are taken together with their intervening atoms to form an optionally substituted ring (e.g., formula I-e).
• a formed ring e.g., Ring A in formula I-e, is 3, 4, 5, 6, 7, 8, 9, or 10-membered.
• a formed ring is 3-membered. In some embodiments, a formed ring is 4-membered. In some embodiments, a formed ring is 5-membered. In some embodiments, a formed ring is 6- membered. In some embodiments, a formed ring is 7-membered. In some embodiments, a formed ring is 8-membered. In some embodiments, a formed ring is 9-membered. In some embodiments, a formed ring is 10-membered. In some embodiments, a formed ring is monocyclic. In some embodiments, a formed ring is bicyclic. In some embodiments, a formed ring is polycyclic. In some embodiments, a formed ring is saturated.
• a formed ring is partially unsaturated. In some embodiments, a formed ring has no heteroatoms. In some embodiments, a formed ring is an optionally substituted 3- membered saturated aliphatic ring. In some embodiments, a formed ring is an optionally substituted 4- membered saturated aliphatic ring. In some embodiments, a formed ring is an optionally substituted 5- membered saturated aliphatic ring. In some embodiments, a formed ring is an optionally substituted 6- membered saturated aliphatic ring. In some embodiments, a formed ring is an optionally substituted 7- membered saturated aliphatic ring.
• a formed ring is an optionally substituted 8- membered saturated aliphatic ring. In some embodiments, a formed ring is an optionally substituted 9- membered saturated aliphatic ring. In some embodiments, a formed ring is an optionally substituted 10- membered saturated aliphatic ring.
• R 3 is -H
• R 1 is optionally substituted Ci- 6 aliphatic or phenyl
• R 5 is optionally substituted Ci- 6 aliphatic
• R 6 is -H.
• R 3 is -H
• R 1 and R 5 are taken together with their intervening atoms to form an optionally substituted ring
• R 6 is -H.
• R 3 is -H
• R 1 and R 5 are taken together with their intervening atoms to form an optionally substituted 5- or 6-membered ring
• R 6 is -H.
• R 3 is -H
• R 1 and R 5 are taken together with their intervening atoms to form an optionally substituted 5-membered saturated ring having no heteroatom in addition to the nitrogen to which R 5 is attached
• R 6 is -H
• R 3 is -H
• R 1 and R 5 are taken together with their intervening atoms to form an optionally substituted 6- membered saturated ring having no heteroatom in addition to the nitrogen to which R 5 is attached
• R 6 is -H.
• a ring formed by R 1 and R 5 taken together are unsubstituted.
• -OH and -N(R 5 )(R 6 ) are trans. In some embodiments, -OH and -N(R 5 )(R 6 ) are cis.
• the carbon to which R 1 and -OH are attached is R. In some embodiments, the carbon to which R 1 and -OH are attached is S.
• R 1 is hydrogen. In some embodiments, R 1 is not hydrogen.
• R 1 is optionally substituted Ci- 6 aliphatic or phenyl. In some embodiments, R 1 is methyl. In some embodiments, R 1 is phenyl.
• R 3 is hydrogen. In some embodiments, R 5 is hydrogen.
• R 5 is not hydrogen. In some embodiments, R 5 is optionally substituted Ci- 6 aliphatic or phenyl. In some embodiments, R 5 is methyl. In some embodiments, R 5 is phenyl. In some embodiments, R 6 is hydrogen. In some embodiments, R 6 is not hydrogen. In some embodiments, as demonstrated by certain example data, compounds with trans -OH and -N(R 5 )(R 6 ) can provide high yields and/or diastereoselectivity. In some embodiments, as demonstrated by certain example data, compounds with trans -OH and -N(R 5 )(R 6 ) can provide both high yields and diastereoselectivity.
• a provided compound e.g., a compound of formula I-e, is selected
• a provided compound is or a salt there of. In some embodiments,
• a provided compound is or a salt there of. In some embodiments, a provided
• a provided compound is or a salt there of. In some embodiments, a provided compound is or a salt there of. In some embodiments, a provided compound is or a salt
• a provided compound is or a salt thereof.
• a provided compound i salt thereof in some embodiments, a provided compound i salt thereof.
• a provided compound has the structure of formula II:
• Ring A is an optionally substituted 3-20 membered monocyclic, bicyclic or polycyclic ring having 0-10 heteroatoms independently selected from oxygen, nitrogen, sulfur, phosphorus and silicon; each of R 1 , R 2 , R 3 , R 4 , and R 5 is independently -H, -L s -R, halogen, -CN, -NO2, -L s -Si(R) 3 , -OR, -SR, or -N(R) 2 ;
• each L s is independently a covalent bond, or a bivalent, optionally substituted, linear or branched group selected from a C i-30 aliphatic group and a C i-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 group selected from Ci- 6 alkylene, Ci- 6 alkenylene, C-C . a bivalent C1-C6 heteroaliphatic group having 1-5 heteroatoms independently selected from oxygen, nitrogen, sulfur, phosphorus and silicon, -C(R’)2-,
• each -Cy- is independently an optionally substituted bivalent group selected from a C3-20 cycloaliphatic ring, a CV20 aryl ring, a 5-20 membered heteroaryl ring having 1-10 heteroatoms independently selected from oxygen, nitrogen, sulfur, phosphorus and silicon, and a 3-20 membered heterocyclyl ring having 1-10 heteroatoms independently selected from oxygen, nitrogen, sulfur, phosphorus and silicon;
• each Cy L is independently an optionally substituted tetravalent group selected from a C3-20 cycloaliphatic ring, a CV20 aryl ring, a 5-20 membered heteroaryl ring having 1-10 heteroatoms independently selected from oxygen, nitrogen, sulfur, phosphorus and silicon, and a 3-20 membered heterocyclyl ring having 1-10 heteroatoms independently selected from oxygen, nitrogen, sulfur, phosphorus and silicon;
• each R’ is independently -R, -C(0)R, -C(0)OR, or -S(0) 2 R;
• t 0-20;
• R 6 is -L-R’ ;
• L is a covalent bond, or optionally substituted Ci- 6 alkylene, wherein one or more methylene units are optionally and independently replaced with -L’-; or L is L s ;
• L’ is a covalent bond, optionally substituted bivalent C1-3 alkylene, -C(R 3 )(R 4 )-,
• each R is independently -H, or an optionally substituted group selected from Ci-30 aliphatic, Ci-30 heteroaliphatic having 1-10 heteroatoms independently selected from oxygen, nitrogen, sulfur, phosphorus and silicon, Ce-30 aryl, Ce-30 arylaliphatic, Ce-30 arylheteroaliphatic having 1-10 heteroatoms independently selected from oxygen, nitrogen, sulfur, phosphorus and silicon, 5-30 membered heteroaryl having 1-10 heteroatoms independently selected from oxygen, nitrogen, sulfur, phosphorus and silicon, and 3-30 membered heterocyclyl having 1-10 heteroatoms independently selected from oxygen, nitrogen, sulfur, phosphorus and silicon, or two R groups are optionally and independently taken together to form a covalent bond, or:
• a provided compound has the structure of formula Il-a:
• a provided compound of structure Il-a has the structure of formula
• a compound of formula Il-a has the structure of formula Il-b.
• a provided compound of structure Il-a has the structure of formula
• a compound of formula Il-a has the structure of formula II-c.
• R 8 is -OH.
• R 6 is -H.
• R 5 is optionally substituted alkyl.
• R 5 is methyl.
• t is 0.
• R 3 is optionally substituted alkyl.
• R 3 is methyl.
• R 3 is optionally substituted phenyl.
• R 3 is phenyl.
• R 3 is optionally substituted C 3-10 cycloalkyl.
• R 3 is optionally substituted cyclohexyl.
• R 3 is cyclohexyl.
• Ring A is an optionally substituted 3-20 membered monocyclic, bicyclic or polycyclic ring having 0-10 heteroatoms independently selected from oxygen, nitrogen, sulfur, phosphorus and silicon.
• Ring A is or comprises at least one monocyclic saturated or partially unsaturated monocyclic ring moiety, optionally as part of a bicyclic or polycyclic system.
• Ring A is monocyclic.
• Ring A is bicyclic or polycyclic comprising at least one monocyclic saturated or partially unsaturated monocyclic ring moiety, and optionally one or more aromatic monocyclic moieties.
• Ring A is or comprises at least one saturated monocyclic ring moiety.
• R 8 is connected to a sp 3 ring atom of Ring A. In some embodiments, R 8 is connected to a sp 3 carbon ring atom of Ring A. In some embodiments, R 3 is connected to a sp 3 ring atom of Ring A. In some embodiments, R 3 is connected to a sp 3 carbon ring atom of Ring A. In some embodiments, -N(R 5 )(R 6 ) is connected to a sp 3 ring atom of Ring A. In some embodiments, -N(R 5 )(R 6 ) is connected to a sp 3 carbon ring atom of Ring A.
• Ring A is optionally substituted C 3-10 cycloalkyl. In some embodiments, Ring A is optionally substituted cyclohexyl. In some embodiments, Ring A is cyclohexyl. In some embodiments, R 8 and -N(R 5 )(R 6 ) are cis. In some embodiments, R 8 and -N(R 5 )(R 6 ) are trans.
• a provided compound of formula II is selected from
• a provided compound is a compound of II-c or a salt thereof.
• R 3 and R 5 are R, and are taken together to form an optionally substituted ring as described in the present disclosure.
• a provided compound of formula II is selected from
• a provided compound has the structure of formula III:
• Ring A’ is an optionally substituted 3-20 membered monocyclic, bicyclic or polycyclic ring having 1-10 heteroatoms independently selected from oxygen, nitrogen, sulfur, phosphorus and silicon, wherein Ring A’ comprises a -N(R 6 )- moiety;
• each of R 1 , R 2 , R 3 and R 4 is independently -H, -L s -R, halogen, -CN, -NO2, -L s -Si(R) 3 , -OR, -SR, or -N(R) 2 ;
• each L s is independently a covalent bond, or a bivalent, optionally substituted, linear or branched group selected from a Ci-30 aliphatic group and a Ci-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 group selected from Ci- 6 alkylene, Ci- 6 alkenylene, CoC _ a bivalent C1-C6 heteroaliphatic group having 1-5 heteroatoms independently selected from oxygen, nitrogen, sulfur, phosphorus and silicon, -C(R’) 2- , -Cy-, -0-, -S-, -S-S-, -N(R’)-, -C(O)-, -C(S)-, -C(NR’)-, -C(0)N(R’)-, -N(R’)C(0)N(R’)-, -N(R’)C(0)N(R
• each -Cy- is independently an optionally substituted bivalent group selected from a C 3-2 o cycloaliphatic ring, a CV20 aryl ring, a 5-20 membered heteroaryl ring having 1-10 heteroatoms independently selected from oxygen, nitrogen, sulfur, phosphorus and silicon, and a 3-20 membered heterocyclyl ring having 1-10 heteroatoms independently selected from oxygen, nitrogen, sulfur, phosphorus and silicon;
• each Cy L is independently an optionally substituted tetravalent group selected from a C 3-2 o cycloaliphatic ring, a CV20 aryl ring, a 5-20 membered heteroaryl ring having 1-10 heteroatoms independently selected from oxygen, nitrogen, sulfur, phosphorus and silicon, and a 3-20 membered heterocyclyl ring having 1-10 heteroatoms independently selected from oxygen, nitrogen, sulfur, phosphorus and silicon;
• each R’ is independently -R, -C(0)R, -C(0)OR, or -S(0) 2 R; t is 0-20;
• R 6 is -L-R’ ;
• R 8 is -L-R 7 , -L-C(R')(R 2 )-R 7 . or -L s -R 7 ;
• R 7 is -OH or -SH
• L is a covalent bond, or optionally substituted Ci- 6 alkylene, wherein one or more methylene units are optionally and independently replaced with -L’-; or L is L s ;
• L’ is a covalent bond, optionally substituted bivalent C1-3 alkylene, -C(R 3 )(R 4 )-,
• each R is independently -H, or an optionally substituted group selected from Ci-30 aliphatic, Ci-30 heteroaliphatic having 1-10 heteroatoms independently selected from oxygen, nitrogen, sulfur, phosphorus and silicon, Ce-30 aryl, Ce-30 arylaliphatic, Ce-30 arylheteroaliphatic having 1-10 heteroatoms independently selected from oxygen, nitrogen, sulfur, phosphorus and silicon, 5-30 membered heteroaryl having 1-10 heteroatoms independently selected from oxygen, nitrogen, sulfur, phosphorus and silicon, and 3-30 membered heterocyclyl having 1-10 heteroatoms independently selected from oxygen, nitrogen, sulfur, phosphorus and silicon, or
• R groups on the same atom are optionally and independently taken together with the atom to form an optionally substituted, 3-30 membered, monocyclic, bicyclic or polycyclic ring having, in addition to the atom, 0-10 heteroatoms independently selected from oxygen, nitrogen, sulfur, phosphorus and silicon; or
• two or more R groups on two or more atoms are optionally and independently taken together with their intervening atoms to form an optionally substituted, 3-30 membered, monocyclic, bicyclic or polycyclic ring having, in addition to the intervening atoms, 0-10 heteroatoms independently selected from oxygen, nitrogen, sulfur, phosphorus and silicon.
• a provided compound has the structure of formula Ill-a:
• a compound of formula III has the structure of formula Ill-a.
• a provided compound has the structure of formula Ill-b:
• a compound of formula Ill-a has the structure of formula III-b.
• R 8 is bonded to a carbon atom (C 2 ) next to the nitrogen atom in -N(R 6 )- (N 1 ) (e.g., formula Ill-a, formula III-b, etc.). In some embodiments, R 8 is bonded to a carbon atom next to C 2 (C 3 ). In some embodiments, R 8 is bonded to a carbon atom next to C 3 that is not C 2 (C 4 ). In some embodiments, R 8 is bonded to a carbon atom next to C 4 which is not C 3 (C 5 ). In some embodiments, R 8 is bonded to a carbon atom next to C 5 which is not C 4 (C 6 ).
• R 8 is -OH. In some embodiments, R 6 is -H. In some embodiments, R 5 is optionally substituted alkyl. In some embodiments, R 5 is methyl. In some embodiments, t is 0. In some embodiments, R 3 is optionally substituted alkyl. In some embodiments, R 3 is methyl. In some embodiments, R 3 is optionally substituted phenyl. In some embodiments, R 3 is phenyl. In some embodiments, R 3 is optionally substituted C3-10 cycloalkyl. In some embodiments, R 3 is optionally substituted cyclohexyl. In some embodiments, R 3 is cyclohexyl.
• Ring A’ is an optionally substituted 3-20 membered monocyclic, bicyclic or polycyclic ring having 1-5 heteroatoms independently selected from oxygen, nitrogen, sulfur, phosphorus and silicon, wherein Ring A’ comprises a -N(R 6 )- moiety.
• Ring A’ is Ring A as described in the present disclosure, wherein Ring A comprises a nitrogen ring atom.
• Ring A’ is or comprises at least one monocyclic saturated or partially unsaturated monocyclic ring moiety, optionally as part of a bicyclic or polycyclic system. In some embodiments, Ring A’ is monocyclic.
• Ring A’ is bicyclic or polycyclic comprising at least one monocyclic saturated or partially unsaturated monocyclic ring moiety, and optionally one or more aromatic monocyclic moieties. In some embodiments, Ring A’ is or comprises at least one saturated monocyclic ring moiety. In some embodiments, R 8 is connected to a sp 3 ring atom of Ring A’. In some embodiments, R 8 is connected to a sp 3 carbon ring atom of Ring A’. In some embodiments, R 3 is connected to a sp 3 ring atom of Ring A’. In some embodiments, R 3 is connected to a sp 3 carbon ring atom of Ring A’ . In some embodiments, the nitrogen to which R 6 is attached is sp 3 .
• a provided compound of formula III is selected from
• a provided compound of formula III is selected from compounds listed in Table 4 below and salts thereof.
• a compound of formula I is a compound of formula I-a. In some embodiments, a compound of formula I is a compound of formula I-a-1. In some embodiments, a compound of formula I is a compound of formula I-a-2. In some embodiments, a compound of formula I is a compound of formula I-b. In some embodiments, a compound of formula I is a compound of formula I-c In some embodiments, a compound of formula I is a compound of formula I-d In some embodiments, a compound of formula I is a compound of formula I-e In some embodiments, a compound of formula I is a compound of formula II. In some embodiments, a compound of formula I is a compound of formula Il-a.
• a compound of formula I is a compound of formula Il-b. In some embodiments, a compound of formula I is a compound of formula II-c. In some embodiments, a compound of formula I is a compound of formula III. In some embodiments, a compound of formula I is a compound of formula Ill-a. In some embodiments, a compound of formula I is a compound of formula Ill-b.
• a provided compound is an enantiomer of a compound selected from Table 1 or a salt thereof. In some embodiments, a provided compound is a diastereomer of a compound selected from Table 1 or a salt thereof.
• a provided compound is an enantiomer of a compound selected from Table 2 or a salt thereof. In some embodiments, a provided compound is a diastereomer of a compound selected from Table 2 or a salt thereof.
• a provided compound is an enantiomer of a compound selected from Table 3 or a salt thereof. In some embodiments, a provided compound is a diastereomer of a compound selected from Table 3 or a salt thereof.
• a provided compound is an enantiomer of a compound selected from Table 4 or a salt thereof. In some embodiments, a provided compound is a diastereomer of a compound selected from Table 4 or a salt thereof.
• a chiral auxiliary can be readily removed by a base (base-labile, e.g., under an anhydrous condition substantially free of water; in many instances, preferably before oligonucleotides comprising intemucleotidic linkages comprising such chiral auxiliaries are exposed to conditions/reagent systems comprising a substantial amount of water, particular in the presence of a base(e.g., cleavage conditions/reagent systems using NH 4 OH)) and provides various advantages as described herein, e.g., high crude purity, high yield, high stereoselectivity, more simplified operation, fewer steps, further reduced manufacture cost, and/or more simplified downstream formulation (e.g., low amount of salt(s) after cleavage), etc.
• base-labile e.g., under an anhydrous condition substantially free of water; in many instances, preferably before oligonucleotides comprising intemucleotidic linkages comprising such chiral auxiliaries are exposed to
• auxiliaries may provide alternative or additional chemical compatibility with other functional and/or protection groups.
• base-labile chiral auxiliaries are particularly useful for construction of chirally controlled non-negatively charged intemucleotidic linkages (e.g., neutral intemucleotidic linkages such as nOOl); in some instances, they can provide significantly improved yield and/or crude purity with high stereoselectivity, e.g., when utilized with removal using a base under an anhydrous condition.
• such a chiral auxiliary is bonded to a linkage phosphorus via an oxygen atom (e.g., which corresponds to a -OH group in a corresponding chiral auxiliary compound, e.g., a compound of formula I), the carbon atom in the chiral auxiliary to which the oxygen is bonded (the alpha carbon) also bonds to -H (in addition to other groups; in some embodiments, a secondary carbon), and the next carbon atom (the beta carbon) in the chiral auxiliary is boned to one or two electron-withdrawing groups.
• R 7 is -OH.
• R 1 is -H.
• R 2 comprises one or two electron-withdrawing groups or can otherwise facilitate remove of the chiral auxiliary by a base.
• R 6 is -H.
• R 1 is -H
• R 2 comprises one or two electron-withdrawing groups
• R 6 is -H
• R 7 is -OH.
• R 1 is -H
• R 2 comprises one or two electron-withdrawing groups
• R 6 is -H
• R 7 is -OH
• a methylene unit of L is replaced with -C(R 3 )(R 4 )-
• R 3 and R 4 is taken together with R 5 to form with their intervening atoms a ring as described herein (e.g., an optionally substituted 3-20 membered monocyclic, bicyclic or polycyclic ring having in addition to the nitrogen atom to which R 5 is on, 0-5 heteroatoms (e.g., an optionally substituted 3, 4, 5, or 6-membered monocyclic saturated ring having in addition to the nitrogen atom to which R 5 is on no other heteroatoms)).
• an electronic -withdrawing group comprises and/or is connected to the carbon atom through, e.g., -S(O)-, — S(0) 2— , -P(0)(R 1 )-, -P(S)R 1 -, or -C(O)-.
• an electron-withdrawing group is -CN, -NO2, halogen, -C(0)R 1 , -C(0)OR’, -C(0)N(R’) 2 , -S(0)R 1 , -S(0) 2 R 1 , -P(W)(R 1 ) 2 , -R(0)( ⁇ ) 2 , -P(0)(OR’) 2 , or -P(S)(R 1 ) 2 .
• an electron-withdrawing group is aryl or heteroaryl, e.g., phenyl, substituted with one or more of -CN, -N0 2 , halogen, C (0)R 1 .
• R 2 is -L-R’.
• R 2 is -L’-L”-R’, wherein L’ is — C(R) 2— or optionally substituted -CH 2- , and L” is -P(0)(R’)-, -P(0)(R’)0-, -P(0)(OR’) , -P(0)(0R’)0- -P(0)[N(R’)]-, -P(0)[N(R’)]0- -P(0)[N(R’)][N(R’)]- -P(S)(R’)- -S(0) 2- -S(0) 2 - -S(0) 2 0- -S(O)-, -C(O)-, -C(0)N(R’)-, or -S-.
• L’ is -C(R) 2 -
• L’ is optionally substituted -CH 2 -.
• L’ is -C(R) 2 -.
• each R is independently hydrogen, or an optionally substituted group selected from Ci-Ce aliphatic, carbocyclyl, aryl, heteroaryl, and heterocyclyl.
• L’ is -CH 2 -.
• L” is -P(0)(R’) , -P(S)(R’) _ , -S(0) 2 -.
• R 2 is -L’-C(0)N(R’) 2 .
• R 2 is -L’-P(0)(R’) 2 .
• R 2 is -L’-P(S)(R’) 2 .
• each R’ is independently optionally substituted aliphatic, heteroaliphatic, aryl, or heteroaryl as described in the present disclosure (e.g., those embodiments described for R).
• each R’ is independently optionally substituted phenyl.
• each R’ is independently optionally substituted phenyl wherein one or more substituents are independently selected from -CN, -OMe, -Cl, -Br, and -F.
• each R’ is independently substituted phenyl wherein one or more substituents are independently selected from -CN, -OMe, -Cl, -Br, and -F. In some embodiments, each R’ is independently substituted phenyl wherein the substituents are independently selected from -CN, -OMe, -Cl, -Br, and -F. In some embodiments, each R’ is independently mono-substituted phenyl, wherein the substituent is independently selected from -CN, -OMe, -Cl, -Br, and -F. In some embodiments, two R’ are the same. In some embodiments, two R’ are different.
• R 2 is -L’-S(0)R ⁇ In some embodiments, R 2 is -L’-C(0)N(R’) 2 . In some embodiments, R 2 is -L’-S(0) 2 R ⁇ In some embodiments, R’ is optionally substituted aliphatic, heteroaliphatic, aryl, or heteroaryl as described in the present disclosure (e.g., those embodiments described for R). In some embodiments, R’ is optionally substituted phenyl. In some embodiments, R’ is optionally substituted phenyl wherein one or more substituents are independently selected from -CN, -OMe, -Cl, -Br, and -F.
• R’ is substituted phenyl wherein one or more substituents are independently selected from -CN, -OMe, -Cl, -Br, and -F. In some embodiments, R’ is substituted phenyl wherein each substituent is independently selected from -CN, -OMe, -Cl, -Br, and -F. In some embodiments, R’ is mono-substituted phenyl. In some embodiments, R’ is mono-substituted phenyl, wherein the substituent is independently selected from -CN, -OMe, -Cl, -Br, and -F. In some embodiments, a substituent is an electron-withdrawing group.
• an electron-withdrawing group is -CN, -N0 2 , halogen, -C(0)R 1 , -C(0)0R’, -C(0)N(R’) 2 , -S(0)R 1 , -S(0) 2 R 1 , -P(W)(R 1 ) 2 , -P(0)(R 1 ) 2 , -P(0)(0R’) 3 ⁇ 4 or -P(S)(R 1 ) 2 .
• R 2 is optionally substituted -CH 2 -L”-R, wherein each of L” and R is independently as described in the present disclosure.
• R 2 is optionally substituted — CH(— L”— R) 2 , wherein each of L” and R is independently as described in the present disclosure.
• R 2 is optionally substituted -CH(-S-R) 2 .
• R 2 is optionally substituted -CH 2- S-R.
• the two R groups are taken together with their intervening atoms to form a ring.
• a formed ring is an optionally substituted 5, 6, 7-membered ring having 0-2 heteroatoms in addition to the intervening heteroatoms.
• R 2 is optionally substituted .
• R 2 is S .
• -S- may be converted to -S(O)- or S(0) 2 - e.g., by oxidation, e.g, t facilitate removal by a base.
• R 2 is -L’-R’, wherein each variable is as described in the present disclosure.
• R 2 is -CHR-R’.
• R 2 is -CH 2- R ⁇
• R 2 is -CH(R’) 2 .
• R 2 is -C(R’)3.
• R’ is optionally substituted aryl or heteroaryl.
• R’ is substituted aryl or heteroaryl wherein one or more substituents are independently an electron-withdrawing group.
• -L’- is optionally substituted -CH 2-
• R’ is R, wherein R is optionally substituted aryl or heteroaryl.
• R is substituted aryl or heteroaryl wherein one or more substituents are independently an electron-withdrawing group.
• R is substituted aryl or heteroaryl wherein each substituent is independently an electron-withdrawing group.
• R is aryl or heteroaryl substituted with two or more substituents, wherein each substituent is independently an electron-withdrawing group.
• an electron-withdrawing group is -CN, -N0 2 , halogen, -CCOJR 1 , -C(0)OR’, -C(0)N(R’) 2 , )(R 1 ) 2 ,
• R’ is /;-N0 2 Ph-. In some embodiments, R’ is . In some embodiments, R’ In some embodiments, R’ is . In some embodiments, R’ In some embodiments, R’ is In some embodiments, R 2 is , . In some embodiments, R’ is In some embodiments, R’ is 2,4,6-trichlorophenyl. In some embodiments, R’ is 2,4,6-trifluorophenyl. In some embodiments, R 2 is -CH(4-chlorophenyl) 2 . In some embodiments, R 2 is
• R is -CH(R’)2 , wherein each Me0 2 S— c " Z
• R 2 is -L’-S(0) 2 R ⁇ wherein each variable is as described in the present disclosure.
• R 2 is -Cfh-SiO ⁇ R’ .
• R 2 is -L’-S(0)R’, wherein each variable is as described in the present disclosure.
• R 2 is -CH 2- S(0)R ⁇
• R 2 is -L’-C(0) 2 R ⁇ wherein each variable is as described in the present disclosure.
• R 2 is -Cfh-CiO ⁇ R’.
• R 2 is -L’-C(0)R’, wherein each variable is as described in the present disclosure.
• R is not phenyl, or mono-, di- or tri-substituted phenyl, wherein each substituent is selected from -NO2, halogen, -CN, Ci-3 alkyl, and C1-3 alkyloxy.
• R is substituted aryl or heteroaryl wherein one or more substituents are independently an electron-withdrawing group.
• R is substituted aryl or heteroaryl wherein each substituent is independently an electron-withdrawing group.
• R is aryl or heteroaryl substituted with two or more substituents, wherein each substituent is independently an electron-withdrawing group.
• an electron- withdrawing group is -CN, -NO2, halogen, -C(0)R 1 , -C(0)0R’, -C(0)N(R’) 2 , -S(0)R 1 , -S(0) 2 R 1 , -P(W)(R 1 )2, -P(0)(R 1 ) 2 , -P(0)(0R’) 2 , or -P(S)(R 1 )2.
• R’ is phenyl.
• R’ is substituted phenyl. In some embodiments, R’ is Cl In some embodiments, R’ is In some embodiments, R’ is optionally substituted Ci- 6 aliphatic. In some embodiments, R’ is t-butyl. In some embodiments, R’ is isopropyl. In some embodiments, R’ is methyl. In some embodiments, R 2 is -CH 2 C(0)0Me. In some embodiments, R 2 is -CH 2 C(0)Ph. In some embodiments, R 2 is -CH 2 C(0)- tBu.
• R 2 is -L -NO2. In some embodiments, R 2 is -CH2-NO2. In some embodiments, R 2 is -L’-S(0) 2 N(R’) 2 . In some embodiments, R 2 is -CH 2- S(0) 2 N(R’) 2 . In some embodiments, R 2 is -L’-S(0) 2 NHR ⁇ In some embodiments, R 2 is -CH 2- S(0) 2 NHR ⁇ In some embodiments, R’ is methyl. In some embodiments, R 2 is -CH 2 -S(0) 2 NH(CH 3 ). In some embodiments, R’ is -CH2PI1. In some embodiments, R 2 is -CH 2 -S(0) 2 NH(CH 2 Ph).
• R 2 is -CH 2 -S(0) 2 N(CH 2 Ph)(0CH 3 ). In some embodiments, R 2 is -CH 2 -S(0) 2 N(CH 2 Ph)(0CH 3 ). In some embodiments, R 2 is -L’-S(0) 2 0R ⁇ In some embodiments, R 2 is -CH 2 -S(0) 2 0R ⁇ In some embodiments, R 2 is -CH 2 -S(0) 2 0Ph. In some embodiments, R 2 is -CH 2 -S(0) 2 0CH 3 . In some embodiments, R 2 is -CH 2 -S(0) 2 0CH 2 Ph.
• R 2 is -L’-P(0)(R’) 2 In some embodiments, R 2 is -CH 2 -P(0)(R’) 2 . In some embodiments, R 2 is -L’-P(0)[N(R’) 2 ] 2 . In some embodiments, R 2 is -CH 2 -P(0)[N(R’) 2 ] 2 . In some embodiments, R 2 is -L’-P(0)[0(R’) 2 ] 2 ⁇ In some embodiments, R 2 is -CH 2 -P(0)[0(R’) 2 ] 2 ⁇ In some embodiments, R 2 is -L’-P(0)(R’)[N(R’) 2 ] 2 .
• R 2 is -CH 2 -C(0)N(R’) 2 .
• each R’ is independently R.
• one R’ is optionally substituted aliphatic, and one R is optionally substituted aryl.
• one R’ is optionally substituted Ci- 6 aliphatic, and one R is optionally substituted phenyl.
• each R’ is independently optionally substituted Ci- 6 aliphatic.
• R 2 is -CH 2 -P(0)(CH 3 )Ph.
• R 2 is -CH 2 -P(0)(CH 3 ) 2 .
• R 2 is -L’-SR’. In some embodiments, R 2 is -Cfh-SR’. In some embodiments, R’ is optionally substituted phenyl. In some embodiments, R’ is phenyl. [00280] In some embodiments, a provided chiral reagent has the structure
• each R 1 is independently as described in the present disclosure.
• each R 1 is independently as described in the present disclosure. In some embodiments, each R 1 is independently R as described in the present disclosure. In some embodiments, each R 1 is independently R, wherein R is optionally substituted aliphatic, aryl, heteroaliphatic, or heteroaryl as described in the present disclosure. In some embodiments, each R 1 is phenyl. In some embodiments, R 1 is -L-R ⁇ In some embodiments, R 1 is -L-R’, wherein L is -0-, -S-, or -N(R’). In some embodiments, a provided chiral reagent has the structure of
• each X 1 is independently -CN, -OR, -Cl, -Br, or -F, wherein R is not -H.
• R is optionally substituted Ci- 6 aliphatic.
• R is optionally substituted Ci- 6 alkyl.
• R is -CH3.
• one or more X 1 are independently electron-withdrawing groups (e.g., -CN, -N0 2 , halogen, -C(0)R 1 , -C(0)OR ⁇ -C(0)N(R’) 2 , -S OJR 1 , -S(0) 2 R 1 , -R( ⁇ n)( ⁇ ) 2 , -P(0)(R 1 ) 2 , -P(0)(OR’) 3 ⁇ 4 -P(S)(R 1 ) 2 , etc ).
• a provided chiral reagent has the structure
• a provided chiral reagent has
• R 1 is R as described in the present disclosure.
• R 1 is R, wherein R is optionally substituted aliphatic, aryl, heteroaliphatic, or heteroaryl as described in the present disclosure.
• R 1 is -L-R ⁇ In some embodiments, R 1 is -L-R’, wherein L is -0-, -S-, or
• X 1 is -H, an electron-withdrawing group, -N0 2 , -CN, -OR, -Cl, -Br, or -F, and W is O or S.
• a provided chiral reagent has the structure wherein
• X 1 is an electron-withdrawing group (e.g., -CN, -NO2, halogen, -C(0)R 1 , -C(0)0R’, -C(0)N(R’) 2 , -SiOJR 1 , -S OfcR 1 , -P(W)(R 1 ) 2 , -P(0)(R 1 ) 2 , -P(0)(0R’)2,-P(S)(R 1 ) 2 , etc.).
• X 1 is an electron-withdrawing group that is not -CN,— N0 2 , or halogen.
• X 1 is not -H, -CN, -N0 2 , halogen, or C1-3 alkyloxy.
• R 22 and R 23 are both R, and the two R groups are taken together with their intervening atoms to form an optionally substituted aryl or heteroaryl ring as described herein.
• one or more substituents are independently electron- withdrawing groups.
• R 21 and R 24 are both R, and the two R groups are taken together with their intervening atoms to form an optionally substituted ring as described herein.

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• 2020
  • 2020-03-19 CN CN202080028682.0A patent/CN113748116A/zh active Pending
  • 2020-03-19 JP JP2021556501A patent/JP2022525541A/ja active Pending
  • 2020-03-19 EP EP20774109.1A patent/EP3941924A4/en active Pending
  • 2020-03-19 SG SG11202110045PA patent/SG11202110045PA/en unknown
  • 2020-03-19 US US17/439,755 patent/US12590115B2/en active Active
  • 2020-03-19 WO PCT/US2020/023735 patent/WO2020191252A1/en not_active Ceased
  • 2020-03-19 MX MX2021011351A patent/MX2021011351A/es unknown
  • 2020-03-19 CA CA3134036A patent/CA3134036A1/en active Pending
  • 2020-03-19 AU AU2020241559A patent/AU2020241559A1/en not_active Abandoned
  • 2020-03-19 KR KR1020217033895A patent/KR20210149750A/ko active Pending
• 2021
  • 2021-09-19 IL IL286487A patent/IL286487A/en unknown
• 2026
  • 2026-03-23 AU AU2026202225A patent/AU2026202225A1/en active Pending

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Non-Patent Citations (1)