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Nucleic acid-based therapeutics that control gene expression have been steadily progressing towards achieving their full clinical potential throughout the last few decades. Rapid progress has been achieved in RNAi-based therapy by optimizing high specificity and gene silencing efficiency using chemically modified siRNAs. Since 2018, four siRNA drug...
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... June-2021, APOLLO-B was enrolled for phase III study of patisiran for the treatment of TTR-mediated Amyloidosis (ATTR) with patients having cardiomyopathy [22]. Similar to patisiran, vutrisiran is another siRNA candidate targeting TTR mRNA and is currently being evaluated in the late stages of phase 3 clinical trial [ Table 1]. ...Similar publications
Silencing genes using small interfering (si) RNA is a promising strategy for treating cancer. However, the curative effect of siRNA is severely constrained by low serum stability and cell membrane permeability. Therefore, improving the delivery efficiency of siRNA for cancer treatment is a research hotspot. Recently, mesoporous silica nanoparticles...
Anti-angiogenic RNAi-based therapy can be considered as a possible strategy for the treatment of endometriosis (EM), which is the most common gynecological disease. Targeted delivery of siRNA therapeutics is a prerequisite for successful treatment without adverse effects. Here we evaluated the RGD1-R6 peptide carrier as a non-viral vehicle for targ...
Macrophages are integral components of the innate immune system, playing a dual role in host defense during infection and pathophysiological states. Macrophages contribute to immune responses and aid in combatting various infections, yet their production of abundant proinflammatory cytokines can lead to uncontrolled inflammation and worsened tissue...
RNA interference (RNAi) therapies have significant potential for the treatment of inflammatory bowel diseases (IBD). Although administering small interfering RNA (siRNA) via an oral route is desirable, various hurdles including physicochemical, mucus, and cellular uptake barriers of the gastrointestinal tract (GIT) impede both the delivery of siRNA...
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... 103 Givosiran, lumasiran, inclisiran, vutrisiran, and nedosiran are all composed of 2 0 -O-Me, 2 0 -F, and PS chemical modifications that enhance stability and reduce immunogenicity. 104 These five drugs are also conjugated to the GalNAc ligand, which was developed to overcome the biological barriers and facilitate targeted delivery. 105 When the bound siRNAs are internalized into hepatocytes through ASGPR-mediated endocytosis, GalNAc siRNAs are rapidly cleaved from the target moiety within acidic endosomes. ...
... 228,229 Furthermore, givosiran, lumasiran, inclisiran, and vutrisiran represent approved GalNAc-siRNA conjugate drugs, which are administered subcutaneously for acute liver porphyria, primary hyperoxaluria type 1, ASCVD, and transthyretin-mediated amyloidosis, respectively. 104 However, the development of naked RNA drug for systemic administration is hampered by RNA stability and rapid renal clearance considerations. 115 The only exception is QPI-1002, a naked siRNA targeting p53 to treat acute kidney injury, 106 in which intravenous administration of QPI-1002 is used to target tubule cells with high renal siRNA concentration. ...
... 115 The only exception is QPI-1002, a naked siRNA targeting p53 to treat acute kidney injury, 106 in which intravenous administration of QPI-1002 is used to target tubule cells with high renal siRNA concentration. 104 ...
G protein-coupled receptors (GPCRs) are the major targets of existing drugs for a plethora of human diseases and dominate the pharmaceutical market. However, over 50% of the GPCRs remain undruggable. To pursue a breakthrough and overcome this situation, there is significant clinical research for developing RNA-based drugs specifically targeting GPCRs, but none has been approved so far. RNA therapeutics represent a unique and promising approach to selectively targeting previously undruggable targets, including undruggable GPCRs. However, the development of RNA therapeutics faces significant challenges in areas of RNA stability and efficient in vivo delivery. This review presents an overview of the advances in RNA therapeutics and the diverse types of nanoparticle RNA delivery systems. It also describes the potential applications of GPCR-targeted RNA drugs for various human diseases.
... 103 Givosiran, lumasiran, inclisiran, vutrisiran, and nedosiran are all composed of 2 0 -O-Me, 2 0 -F, and PS chemical modifications that enhance stability and reduce immunogenicity. 104 These five drugs are also conjugated to the GalNAc ligand, which was developed to overcome the biological barriers and facilitate targeted delivery. 105 When the bound siRNAs are internalized into hepatocytes through ASGPR-mediated endocytosis, GalNAc siRNAs are rapidly cleaved from the target moiety within acidic endosomes. ...
... 228,229 Furthermore, givosiran, lumasiran, inclisiran, and vutrisiran represent approved GalNAc-siRNA conjugate drugs, which are administered subcutaneously for acute liver porphyria, primary hyperoxaluria type 1, ASCVD, and transthyretin-mediated amyloidosis, respectively. 104 However, the development of naked RNA drug for systemic administration is hampered by RNA stability and rapid renal clearance considerations. 115 The only exception is QPI-1002, a naked siRNA targeting p53 to treat acute kidney injury, 106 in which intravenous administration of QPI-1002 is used to target tubule cells with high renal siRNA concentration. ...
... 115 The only exception is QPI-1002, a naked siRNA targeting p53 to treat acute kidney injury, 106 in which intravenous administration of QPI-1002 is used to target tubule cells with high renal siRNA concentration. 104 ...
G protein-coupled receptors (GPCRs) are the major targets of existing drugs for a plethora of human diseases and dominate the pharmaceutical market. However, over 50% of the GPCRs remain undruggable. To pursue a breakthrough and overcome this situation, there is significant clinical research for developing RNA-based drugs specifically targeting GPCRs, but none has been approved so far. RNA therapeutics represent a unique and promising approach to selectively targeting previously undruggable targets, including undruggable GPCRs. However, the development of RNA therapeutics faces significant challenges in areas of RNA stability and efficient in vivo delivery. This review presents an overview of the advances in RNA therapeutics and the diverse types of nanoparticle RNA delivery systems. It also describes the potential applications of GPCR-targeted RNA drugs for various human diseases.
... Once siRNA molecules have successfully penetrated the skin, they may encounter endogenous endonuclease enzymes, which are abundant in the skin, and undergo a degradation process that reduces their therapeutic efficacy [35]. The development of siRNA with improved stability, such as chemically modified siRNAs (including modifications to the sugar ribose of the siRNA, addition of methyl or thiophosphate groups to the siRNA phosphate group, and modification of the siRNA nucleobase), is crucial to protect the molecule from nuclease degradation and increase its halflife in the body [4,[36][37][38]. ...
... A 1 mm 2 region around the injection site was selected for quantification. Colors were deconvoluted using 'H DAB', and the number of nuclei was counted by inverting the nuclei image color, applying median blur ( 3 ), and find maxima ( 30 ). The number of EGFP-positive cells was counted by inverting the DAB image color, applying Gaussian blur ( 2 ), setting the threshold to remove the background counting, and applying watershed, followed by analyzing particles (200 − infinity). ...
Guide RNAs offer programmability for CRISPR-Cas9 genome editing but also add challenges for delivery. Chemical modification, which has been key to the success of oligonucleotide therapeutics, can enhance the stability, distribution, cellular uptake, and safety of nucleic acids. Previously, we engineered heavily and fully modified SpyCas9 crRNA and tracrRNA, which showed enhanced stability and retained activity when delivered to cultured cells in the form of the ribonucleoprotein complex. In this study, we report that a short, fully stabilized oligonucleotide (a ‘protecting oligo’), which can be displaced by tracrRNA annealing, can significantly enhance the potency and stability of a heavily modified crRNA. Furthermore, protecting oligos allow various bioconjugates to be appended, thereby improving cellular uptake and biodistribution of crRNA in vivo. Finally, we achieved in vivo genome editing in adult mouse liver and central nervous system via co-delivery of unformulated, chemically modified crRNAs with protecting oligos and AAV vectors that express tracrRNA and either SpyCas9 or a base editor derivative. Our proof-of-concept establishment of AAV/crRNA co-delivery offers a route towards transient editing activity, target multiplexing, guide redosing, and vector inactivation.
... Unlike single-stranded ribozymes, EGSs, and U1 adapters, double-stranded siRNAs are more resistant to RNases. In addition, their stability and efficiency can be increased by nucleotide modification [24]. To date, more than 30 clinical trials of siRNA-based drugs have been initiated. ...
... CPPs often have the amphipathic nature, i.e., one part of the peptide is hydrophobic and the other is hydrophilic. CPPs not only penetrate into the cell, but can also transport various therapeutic agents (for example, nucleic acids [24]) to the cytoplasm [35,36]. ...
Currently, nucleic acid therapeutics are actively developed for the treatment and prophylactic of metabolic disorders and oncological, inflammatory, and infectious diseases. A growing number of approved nucleic acid-based drugs evidences a high potential of gene therapy in medicine. Therapeutic nucleic acids act in the cytoplasm, which makes the plasma membrane the main barrier for the penetration of nucleic acid-based drugs into the cell and requires development of special vehicles for their intracellular delivery. The optimal carrier should not only facilitate internalization of nucleic acids, but also exhibit no toxic effects, ensure stabilization of the cargo molecules, and be suitable for a large-scale and low-cost production. Cell-penetrating peptides (CPPs), which match all these requirements, were found to be efficient and low-toxic carriers of nucleic acids. CPPs are typically basic peptides with a positive charge at physiological pH that can form nanostructures with negatively charged nucleic acids. The prospects of CPPs as vehicles for the delivery of therapeutic nucleic acids have been demonstrated in numerous preclinical studies. Some CPP-based drugs had successfully passed clinical trials and were implemented into medical practice. In this review, we described different types of therapeutic nucleic acids and summarized the data on the use of CPPs for their intracellular delivery, as well as discussed, the mechanisms of CPP uptake by the cells, as understanding of these mechanisms can significantly accelerate the development of new gene therapy approaches.
... The following options exist for siRNA drug design: delivery of either siRNA precursors or mature siRNA [90]. siRNA precursors are usually 26-28 bp long and have a hairpin-shaped region corresponding to the 5 -end of the leading (antisense) strand [116]. The mature siRNAs form dsRNA complexes of 21 and 23 nt long with a 2-nt unpaired region at the 3 end. ...
Epigenetic therapy is a promising tool for the treatment of a wide range of diseases. Several fundamental epigenetic approaches have been proposed. Firstly, the use of small molecules as epigenetic effectors, as the most developed pharmacological method, has contributed to the introduction of a number of drugs into clinical practice. Secondly, various innovative epigenetic approaches based on dCas9 and the use of small non-coding RNAs as therapeutic agents are also under extensive research. In this review, we present the current state of research in the field of epigenetic therapy, considering the prospects for its application and possible limitations.
... Some common groups added include phosphorothioate (PS), 2'-O-methyl (2'-OMe), 2'-fluoro (2'-F), and 2'-O-methyloxyethyl (2'-MOE) groups. These specific modifications to siRNA and ASO therapeutics allow them to be resistant to nuclease degradation Gangopadhyay and Gore, 2022) and to reduce non-specific effects (Yoo, 2004). ...
... Addition of this moiety is also advantageous because it eliminates the need for premedication . Vutrisiran has 2'-OMe, 2'-F, and PS modifications (Fig. 3) dispersed throughout the chemical structure to increase specificity and potency (Gangopadhyay and Gore, 2022). Like inotersen and patisiran, vutrisiran also binds to the 3'-UTR region of the TTR mRNA (Fig. 4). ...
Systemic diseases of liver origin (SDLO) are complex diseases in multiple organ systems, such as cardiovascular, musculoskeletal, endocrine, renal, respiratory, and sensory organ systems, caused by irregular liver metabolism and production of functional factors. Examples of such diseases discussed in this article include primary hyperoxaluria, familial hypercholesterolemia, acute hepatic porphyria, hereditary transthyretin amyloidosis, hemophilia, atherosclerotic cardiovascular diseases, alpha-1 antitrypsin deficiency-associated liver disease, and complement-mediated diseases. Nucleic acid therapeutics use nucleic acids and related compounds as therapeutic agents to alter gene expression for therapeutic purposes. The two most promising fastest-growing classes of nucleic acid therapeutics are antisense oligonucleotides (ASOs) and small interfering RNAs (siRNAs). For each listed SDLO disease, this article discusses epidemiology, symptoms, genetic causes, current treatment options, and advantages and disadvantages of nucleic acid therapeutics by either ASO or siRNA drugs approved or under development. Furthermore, challenges and future perspectives on adverse drug reactions and toxicity of ASO and siRNA drugs for the treatment of SDLO diseases are also discussed. In summary, this review article will highlight the clinical advantages of nucleic acid therapeutics in targeting the liver for the treatment of SDLO diseases. Significance Statement SDLO diseases contain both rare and common complex diseases caused by irregular functions of the liver. Nucleic acid therapeutics have shown promising clinical advantages to treat SDLO diseases. This article aims to provide the most updated information on targeting the liver with ASO and siRNA drugs. The generated knowledge may stimulate further investigations in this growing field of new therapeutic entities for the treatment of some human systemic diseases, which currently have no or limited options for treatment.
... [7,8] Electronegative substituents at the C2'-position of ribofuranose such as 2'-OMe, 2'-F, and 2'-O-MOE lock the sugar conformation in the C3'-endo (North) state, [9][10][11] that have been extensively incorporated in the clinically approved siRNA drugs and other RNAi therapeutics. [12,13] Moreover, several destabilizing modifications such as glycol nucleic acids (GNA), unlocked nucleic acids (UNA), etc. have been incorporated at the seed region of siRNA to mitigate the off-target mediated toxicity in siRNA drugs. [14,15] Natural RNA modifications including common methylated derivatives such as N 6 -methyl adenosine, 5-methylcytidine, N 3 -methylcytidine, N 3 -methyluridine, N 1 -methylguanidine, and 2'-OMe have numerous biological functions, including regulating gene expression and cellular processes. ...
... Compound 7 was methylated using MeI and K 2 CO 3 in a mixture of acetone and DMF (1 : 1) to obtain the 2'-F-N 3 -methyluridine 8 in 98 % yield. All of these compounds were characterized by 1 H, 13 C NMR and HRMS. NMR-based conformational analysis. ...
... Solvents were removed under reduced pressure using a rotary evaporator maintaining a water-bath temperature of~40°C and a maximum vacuum of 0 mbar. All 1 H, 13 C, 19 F, and HMBC NMR spectra were recorded at room temperature either on a Bruker 400 or a 500 MHz spectrometer. All NMR chemical shift values (δ) are quoted in parts per million (ppm) downfield from TMS and calibrated on the solvent signal. ...
Dual modifications (base and sugar) in siRNA have the potential to overcome off‐target effects and impart nuclease resistance. Here we present the synthesis and detailed structural analysis of N³‐methyluridine and 2’‐alkoxy/fluoro‐N³‐methyluridine modifications, the rationally designed modifications for oligonucleotide therapeutics. Using X‐ray crystallography and ¹H NMR spectral data, it is clear that the C2’‐endo conformation of N³‐methyluridine switches to the C3’‐endo pucker state preferentially due to the introduction of electron‐withdrawing 2’‐alkoxy or fluoro substituents. DFT study confirms that the modifications preferentially acquire the C3’‐endo conformation. NBO analysis indicates that in the presence of the 2’‐alkoxy/fluoro substituents, the C3’‐endo conformation is stabilized due to nO4’→σ*C1’‐N1 anomeric as well as σC3’‐H→σ*C2’‐R and σC1’‐C2’→σ*C3’‐OH hyperconjugation effects. The W−C hydrogen bond energies of the modified nucleosides were calculated using the DFT and SAPT methods at the modified base pair regions, and it was observed that one of the hydrogen bonds gets disrupted due to the presence of the N³‐methyl group, causing significant decline in hydrogen bond energy. These detailed structural studies will help to expand the scope of these destabilizing modifications in the rational design of RNAi‐based therapeutics.
... 41 Givosiran is another siRNA drug that targets δ-aminolevulinate synthase 1 (ALAS1) and is used to treat acute intermittent porphyria. 42 Inclisiran, on the other hand, targets the PCSK9 gene and is used to treat hypercholesterolemia. Lumasiran is an siRNA drug that targets the glycolate oxidase (GO) gene and is used to treat primary hyperoxaluria type 1. 42 miRNAs are small single-strand RNA molecules that are approximately 20-22 nucleotides in length. miRNAs are synthesized by endogenous genes within cells. ...
... 41 Givosiran is another siRNA drug that targets δ-aminolevulinate synthase 1 (ALAS1) and is used to treat acute intermittent porphyria. 42 Inclisiran, on the other hand, targets the PCSK9 gene and is used to treat hypercholesterolemia. Lumasiran is an siRNA drug that targets the glycolate oxidase (GO) gene and is used to treat primary hyperoxaluria type 1. 42 miRNAs are small single-strand RNA molecules that are approximately 20-22 nucleotides in length. miRNAs are synthesized by endogenous genes within cells. ...
Genetic kidney disease is the main cause of chronic kidney disease in children. While the pathogenic genes associated with most genetic kidney diseases have been identified, the underlying mechanisms of disease initiation remain ambiguous, and effective treatment modalities are limited. Gene therapy has emerged as a promising approach for treating genetic diseases. Several gene therapy drugs have been successfully launched to treat genetic diseases affecting the eyes and muscles. However, the development of gene therapy agents for kidney diseases lags behind that of other genetic disorders. In this review, we first comprehensively summarize the main gene therapy strategies. Next, we provide an overview of current treatment options as well as the latest research on gene therapy approaches for pediatric genetic kidney diseases with a particular emphasis on Alport syndrome and polycystic kidney disease. Finally, we discuss the challenges and potential solutions for gene therapy of pediatric genetic kidney diseases.
... Nevertheless, a handful of companies including Alnylam, Ionis, Dicerna, Moderna, BioNtech, and others persisted and eventually developed selective delivery approaches for targeting hepatic diseases or as vaccines [3][4][5]. Concomitantly, novel modifications of RNA backbone structures were devised that both enhanced efficacy and reduced immune recognition [6][7][8][9][10][11]. ...
... How much damage might occur to a single nanostructure over time as it transits endolysosomal pathways is difficult to define experimentally, but the general assumption is that the process of condensation of peptide and nucleotide protects the complex against nucleases and proteases, as has been reported for many of the designs when exposed to serum or concentrated nucleases (e.g., p5RHH [51,52,66]). Furthermore, a now standard array of RNA chemical modifications affords additional longevity to the nucleotide cargo [6][7][8][9][10]. In most cases, the strategy for PBN design is to create a complex that employs the peptide excipient itself as the endosomal permeabilizing entity in an effort to promote rapid escape. ...
Peptide-based nanoparticles (PBN) for nucleotide complexation and targeting of extrahepatic diseases are gaining recognition as potent pharmaceutical vehicles for fine-tuned control of protein production (up- and/or down-regulation) and for gene delivery. Herein, we review the principles and mechanisms underpinning self-assembled formation of PBN, cellular uptake, endosomal release, and delivery to extrahepatic disease sites after systemic administration. Selected examples of PBN that have demonstrated recent proof of concept in disease models in vivo are summarized to offer the reader a comparative view of the field and the possibilities for clinical application.
