Gene Silencing Activity and Hepatic Accumulation of Antisense Oligonucleotides Bearing Cholesterol-Conjugated Thiono Triester at the Gap Region

Nakajima, Mado; Kasuya, Tadashi; Yokota, Shinnichi; Onishi, Reina; Ikehara, Tatsuya; Kugimiya, Akira; Watanabe, Asako

doi:10.1089/nat.2017.0669

Cited by 10 publications

(8 citation statements)

References 23 publications

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“…In the synthesis of these biomolecular hybrids, in which even the bis-conjugation of ONs is needed, orthogonal ligation chemistries play a central role. It is beneficial if the conjugation itself creates a linker that is cleavable [32][33][34]. The linker should also provide efficient conjugation, be stable in physiological conditions, and release the therapeutic ON cargo in appropriate intracellular compartments.…”

Section: Introductionmentioning

confidence: 99%

Expanding the Scope of the Cleavable N-(Methoxy)oxazolidine Linker for the Synthesis of Oligonucleotide Conjugates

et al. 2021

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Oligonucleotides modified by a 2′-deoxy-2′-(N-methoxyamino) ribonucleotide react readily with aldehydes in slightly acidic conditions to yield the corresponding N-(methoxy)oxazolidine-linked oligonucleotide-conjugates. The reaction is reversible and dynamic in slightly acidic conditions, while the products are virtually stable above pH 7, where the reaction is in a ‘‘switched off-state’’. Small molecular examinations have demonstrated that aldehyde constituents affect the cleavage rate of the N-(methoxy)oxazolidine-linkage. This can be utilized to adjust the stability of this pH-responsive cleavable linker for drug delivery applications. In the present study, Fmoc-β-Ala-H was immobilized to a serine-modified ChemMatrix resin and used for the automated assembly of two peptidealdehydes and one aldehyde-modified peptide nucleic acid (PNA). In addition, a triantennary N-acetyl-d-galactosamine-cluster with a β-Ala-H unit has been synthesized. These aldehydes were conjugated via N-(methoxy)oxazolidine-linkage to therapeutically relevant oligonucleotide phosphorothioates and one DNA-aptamer in 19–47% isolated yields. The cleavage rates of the conjugates were studied in slightly acidic conditions. In addition to the diverse set of conjugates synthesized, these experiments and a comparison to published data demonstrate that the simple conversion of Gly-H to β-Ala-H residue resulted in a faster cleavage of the N-(methoxy)oxazolidine-linker at pH 5, being comparable (T0.5 ca 7 h) to hydrazone-based structures.

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Section: Introductionmentioning

confidence: 99%

Expanding the Scope of the Cleavable N-(Methoxy)oxazolidine Linker for the Synthesis of Oligonucleotide Conjugates

et al. 2021

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“…Poor cellular uptake of oligonucleotides having therapeutic potential is still a major problem to be solved. Conjugation of oligoribonucleotides to hydrophobic moieties and their analogs has been reported to enhance their cellular uptake (examples are numerous; see, for instance, [ 5 , 65 , 66 , 67 , 68 , 69 ]). A series of 2′-cholesterol-functionalized 21-mer oligonucleotides 8b – 10b ( Table 1 and Table S2 ) corresponding to the sense strand of a model siRNA-targeting GFP mRNA [ 70 ] was synthesized as described above ( Table 3 and Supplementary Materials, Section 1.9, page 9 ).…”

Section: Resultsmentioning

confidence: 99%

Postsynthetic On-Column 2′ Functionalization of RNA by Convenient Versatile Method

Krasheninina

Fishman

Lomzov

et al. 2020

IJMS

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We report a universal straightforward strategy for the chemical synthesis of modified oligoribonucleotides containing functional groups of different structures at the 2’ position of ribose. The on-column synthetic concept is based on the incorporation of two types of commercial nucleotide phosphoramidites containing orthogonal 2’-O-protecting groups, namely 2’-O-thiomorpholine-carbothioate (TC, as “permanent”) and 2’-O-tert-butyl(dimethyl)silyl (tBDMS, as “temporary”), to RNA during solid-phase synthesis. Subsequently, the support-bound RNA undergoes selective deprotection and follows postsynthetic 2’ functionalization of the naked hydroxyl group. This convenient method to tailor RNA, utilizing the advantages of solid phase approaches, gives an opportunity to introduce site-specifically a wide range of linkers and functional groups. By this strategy, a series of RNAs containing diverse 2’ functionalities were synthesized and studied with respect to their physicochemical properties.

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“…Cholesterol is conjugated to oligos at different locations, including the 5', 3', and middle part of the oligos; importantly, the spacer selection between the ASO and cholesterol moiety is thought critical for biological activity. [123][124][125] Cholesterol-conjugated ASOs are recognized by high and low-affinity lipoproteins and can thus escape renal clearance extending the time the ASO remains in the circulation. They have also been shown to improve cellular uptake of oligos as well as endosomal escape.…”

Section: Delivery and Pharmacokinetics Of Asosmentioning

confidence: 99%

“…Another commonly used method for the conjugation of ASOs, particularly siRNAs, is the use of cholesterol derivatives. Cholesterol is conjugated to oligos at different locations, including the 5', 3', and middle part of the oligos; importantly, the spacer selection between the ASO and cholesterol moiety is thought critical for biological activity 123–125 . Cholesterol‐conjugated ASOs are recognized by high and low‐affinity lipoproteins and can thus escape renal clearance extending the time the ASO remains in the circulation.…”

Section: Delivery and Pharmacokinetics Of Asosmentioning

confidence: 99%

Therapeutic antisense oligonucleotides for movement disorders

Doxakis

2020

Medicinal Research Reviews

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Movement disorders are a group of neurological conditions characterized by abnormalities of movement and posture. They are broadly divided into akinetic and hyperkinetic syndromes. Until now, no effective symptomatic or disease-modifying therapies have been available. However, since many of these disorders are monogenic or have some well-defined genetic component, they represent strong candidates for antisense oligonucleotide (ASO) therapies. ASO therapies are based on the use of short synthetic single-stranded ASOs that bind to disease-related target RNAs via Watson-Crick base-pairing and pleiotropically modulate their function. With information arising from the RNA sequence alone, it is possible to design ASOs that not only alter the expression levels but also the splicing defects of any protein, far exceeding the intervention repertoire of traditional small molecule approaches. Following the regulatory approval of ASO therapies for spinal muscular atrophy and Duchenne muscular dystrophy in 2016, there has been tremendous momentum in testing such therapies for other neurological disorders. This review article initially focuses on the chemical modifications aimed at improving ASO effectiveness, the mechanisms by which ASOs can interfere with RNA function, delivery systems and pharmacokinetics, and the common set of toxicities associated with their application. It, then, describes the pathophysiology and the latest information on preclinical and clinical

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Gene Silencing Activity and Hepatic Accumulation of Antisense Oligonucleotides Bearing Cholesterol-Conjugated Thiono Triester at the Gap Region

Cited by 10 publications

References 23 publications

Expanding the Scope of the Cleavable N-(Methoxy)oxazolidine Linker for the Synthesis of Oligonucleotide Conjugates

Expanding the Scope of the Cleavable N-(Methoxy)oxazolidine Linker for the Synthesis of Oligonucleotide Conjugates

Postsynthetic On-Column 2′ Functionalization of RNA by Convenient Versatile Method

Therapeutic antisense oligonucleotides for movement disorders

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