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WO2024123010A1 - Plateforme luminescente pour la double mesure de l'activité du vhc/mir-122 et utilisation d'une substance rigosertib dérivée pour le traitement contre le vhc résistant au sofosbuvir - Google Patents

Plateforme luminescente pour la double mesure de l'activité du vhc/mir-122 et utilisation d'une substance rigosertib dérivée pour le traitement contre le vhc résistant au sofosbuvir Download PDF

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WO2024123010A1
WO2024123010A1 PCT/KR2023/019800 KR2023019800W WO2024123010A1 WO 2024123010 A1 WO2024123010 A1 WO 2024123010A1 KR 2023019800 W KR2023019800 W KR 2023019800W WO 2024123010 A1 WO2024123010 A1 WO 2024123010A1
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hcv
mir
platform
sofosbuvir
resistant
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English (en)
Korean (ko)
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김종헌
서유나
조성찬
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National Cancer Center Japan
National Cancer Center Korea
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National Cancer Center Japan
National Cancer Center Korea
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Priority claimed from KR1020230173335A external-priority patent/KR20240087568A/ko
Publication of WO2024123010A1 publication Critical patent/WO2024123010A1/fr
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/185—Acids; Anhydrides, halides or salts thereof, e.g. sulfur acids, imidic, hydrazonic or hydroximic acids
    • A61K31/19—Carboxylic acids, e.g. valproic acid
    • A61K31/195—Carboxylic acids, e.g. valproic acid having an amino group
    • A61K31/197—Carboxylic acids, e.g. valproic acid having an amino group the amino and the carboxyl groups being attached to the same acyclic carbon chain, e.g. gamma-aminobutyric acid [GABA], beta-alanine, epsilon-aminocaproic acid or pantothenic acid
    • A61K31/198—Alpha-amino acids, e.g. alanine or edetic acid [EDTA]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12—Antivirals
    • A61P31/14—Antivirals for RNA viruses

Definitions

  • the present invention relates to a luminescent platform for dual measurement of HCV/miR-122 activity and the use of the derivative ligosertip for the treatment of sofosbuvir-resistant HCV, and more specifically, to a dual detection system for screening miR-122 target substances for inhibiting HCV proliferation. It relates to a pharmaceutical composition for preventing or treating sofosbuvir-resistant HCV, including a dual-sensing system and ligosertip.
  • Hepatitis C virus is an RNA virus classified into the Hepacivirus genus of the Flaviviridae family and was discovered in the United States in 1989. This is a clinically very important virus that causes hepatitis, cirrhosis, and liver cancer (HCC), and in developed countries such as the United States and Europe, HCV is the main cause of chronic hepatitis. According to WHO, hepatitis C virus (HCV) is emerging as a more serious problem as approximately 200 million people, or more than 3% of the total population, are infected worldwide, and the number of additional infections is increasing by 3-4 million each year. .
  • DAAs direct-acting antivirals
  • miR-122 is liver-specific and is highly abundant in hepatocytes, with more than 60,000 copies per cell.
  • miR-122 is known to promote adipogenesis, tumor suppression, and immune responses, but the effects of MiR-122 binding to the HCV genome are in stark contrast to the standard mode of action of miRNAs. Binding of miR-122 to the microRNA-122 (miR-122) target site of the HCV genome increases HCV genome translation and replication initiation, and binding of miR-122 obscures the 5' terminal region of HCV RNA, making it immune to host ribonuclease.
  • the present invention in order to select substances that can specifically inhibit HCV proliferation by miR-122, we developed a dual sensing system for screening inhibitors of hepatitis C virus proliferation, and furthermore, we developed a dual sensing system for screening inhibitors of hepatitis C virus proliferation ( We developed a sensing system for screening treatment materials for Beer-resistant HCV).
  • the ligosertib selected using the system of the present invention showed strong antiviral activity in RAS-positive hepatitis C (sofosbuvir-resistant HCV), and the present invention was completed.
  • the purpose of the present invention is to provide a dual-measurement luminescent platform for selecting a target substance for miR-122 to inhibit HCV proliferation.
  • Another object of the present invention is to provide a luminescent platform for screening sofosbuvir-resistant HCV therapeutic substances.
  • Another object of the present invention is to provide a pharmaceutical composition for preventing or treating sofosbuvir-resistant HCV, including ligosertip.
  • the present invention provides a luminescent platform for measuring HCV proliferation, which is composed of a miR-122 binding portion, a Renilla luciferase gene, and an HCV replicon, and is represented by the RNA base sequence of SEQ ID NO: 1; and
  • a miR-122 target material for inhibiting HCV proliferation consisting of a Firefly luciferase gene and a miR-122 binding site, and a miR-122 measurement luminescent platform represented by the RNA base sequence of SEQ ID NO: 2. Provides a dual measurement luminescence platform for screening.
  • HCV replicon replication is reduced in the presence of the target substance, miR-122, for inhibiting HCV proliferation, and the signal of the HCV proliferation measurement luminescent platform is reduced.
  • the signal of the miR-122 instrumented luminescent platform can be increased.
  • the present invention provides a vector containing a dual-measurement luminescent platform for selecting the target substance for miR-122 for inhibiting HCV proliferation.
  • the present invention provides a dual-measurement luminescent platform for selecting a target substance for miR-122 for inhibiting HCV proliferation or a recombinant cell transformed with a vector containing the same.
  • the recombinant cells may be liver cancer-derived cells.
  • the present invention provides a method for selecting a target substance for miR-122 for inhibiting HCV proliferation using a dual-measurement luminescent platform for selecting a target substance for the inhibition of HCV proliferation.
  • the selection method includes (a) a dual-measurement luminescent platform for selecting a miR-122 target material for inhibiting HCV proliferation, or a miR-122 target candidate in recombinant cells transformed with a vector containing the same. processing the material; and
  • the present invention is a luminescent platform for screening sofosbuvir-resistant HCV therapeutic agents, which consists of a miR-122 binding portion, a reporter gene, and an HCV replicon, and is represented by the RNA base sequence of SEQ ID NO: 3 or SEQ ID NO: 4. provides.
  • the reporter gene may be the Renilla luciferase gene or the Firefly luciferase gene.
  • the light-emitting platform is composed of a Firefly luciferase gene and a miR-122 binding site, and is represented by the base sequence of SEQ ID NO: 2. may additionally be included.
  • the present invention provides a vector containing a luminescent platform for screening sofosbuvir-resistant HCV therapeutic substances.
  • the present invention provides a light-emitting platform for selecting a sofosbuvir-resistant HCV therapeutic agent or a recombinant cell transformed with a vector containing the same.
  • the present invention provides a method for screening sofosbuvir-resistant HCV therapeutic substances using the luminescent platform for screening sofosbuvir-resistant HCV therapeutic substances.
  • the selection method is (a) treating a sofosbuvir-resistant HCV treatment candidate to a recombinant cell transformed with a luminescent platform for selecting a sofosbuvir-resistant HCV treatment material or a vector containing the same. steps; and
  • the present invention provides a pharmaceutical composition for preventing or treating sofosbuvir-resistant HCV, including ligosertip.
  • the present invention provides information on treatment of hepatitis C virus resistant to sofosbuvir, including providing information on predicting the effect of administering rigosertib to patients with hepatitis C virus infection showing resistance to sofosbuvir. Provides a method of provision.
  • the dual measurement luminescence platform for selecting a target substance for miR-122 for inhibiting HCV proliferation of the present invention reduces HCV replicon replication in the presence of a target substance for inhibiting HCV proliferation. Since it was confirmed that the signal decreases and the signal of the miR-122 measurement luminescence platform increases, the HCV/miR-122 activity dual measurement luminescence platform of the present invention can be effectively applied to select miR-122 target substances for inhibiting HCV proliferation. .
  • ligosertib derived using the HCV/miR-122 active luminescence platform in which the NS5B region of the HCV replicon is substituted with S282T, is a potent antibacterial agent against RAS-positive hepatitis C virus (sofosbuvir-resistant HCV). Since it was confirmed that it exhibits viral activity, the platform of the present invention can be easily applied to the selection of sofosbuvir-resistant HCV treatments.
  • Figure 1 is a schematic diagram showing the dual measurement luminescent platform (HIRL-2Aneo-NSrep/FL-5 ⁇ 122) for selecting a target substance for miR-122 for inhibiting HCV proliferation of the present invention.
  • Figure 2 is data confirming the degree of Rluc. (Renilla luciferase) and Fluc. (firefly luciferase) luminescence when rigosertib was treated with the dual measurement luminescence platform of the present invention.
  • Figure 3 shows (A) data monitoring the levels of NS5A and ELAVL1/HuR proteins through Western blotting when rigosertib was treated with the dual-measurement luminescence platform of the present invention, and (B) the level of miR-122. This is data analyzed by qRT-PCR for expression.
  • FIG. 4 is a schematic diagram showing (A) a luminescent platform (HIRL-2Aneo-NSrep-5B(S282T)) for screening therapeutic substances for RAS-positive hepatitis C virus (sofosbuvir-resistant HCV) of the present invention, and sofosbuvir-resistant HCV.
  • This data confirms the degree of HCV proliferation through Rluc (Renilla luciferase) luminescence when (B) rigosertib and (C) sofosbuvir were treated on a luminescent platform for selecting therapeutic substances.
  • FIG. 5 is a schematic diagram showing (A) a luminescent platform (HIFL-NSrep-5B(S282T)) for screening a therapeutic material for RAS-positive hepatitis C virus (sofosbuvir-resistant HCV) of the present invention, and a sofosbuvir-resistant HCV therapeutic material.
  • This data confirms the degree of HCV proliferation through Fluc (firefly luciferase) emission when (B) rigosertib and (C) sofosbuvir were treated on the luminescent platform for selection.
  • the present invention is consistent with the present invention, which consists of a miR-122 binding portion, a Renilla luciferase (Rluc) gene, and an HCV replicon, and is expressed as an RNA base sequence of SEQ ID NO: 1. Instrumentation luminescence platform; and
  • MiR-122 for inhibiting HCV proliferation consisting of the Firefly luciferase (Fluc) gene and the miR-122 binding site, and the miR-122 measurement luminescence platform represented by the RNA base sequence of SEQ ID NO: 2 This relates to a dual measurement luminescence platform for target material selection.
  • Fluc Firefly luciferase
  • the dual measurement luminescence platform reduces HCV replicon replication in the presence of a target substance for the inhibition of HCV proliferation, miR-122, thereby reducing the signal of the HCV proliferation measurement luminescence platform and emitting light for measuring the miR-122.
  • the platform's signal may increase.
  • the Renilla luciferase gene or the Firefly luciferase gene was used as the reporter gene, but if necessary, reporter protein genes known in the art can be applied to the present invention.
  • the reporter gene is a group consisting of a green fluorescent protein gene, a red fluorescent protein gene, and a secreted alkaline phosphatase (SeAP) gene. It may be any one or more selected from .
  • the present invention relates to a vector containing a dual-measurement luminescent platform for selecting the target substance for the miR-122 for inhibiting HCV proliferation.
  • the present invention relates to a dual-measurement luminescence platform for selecting a target substance for the miR-122 for inhibiting HCV proliferation or to a recombinant cell transformed with a vector containing the same.
  • expression vector is a gene product containing essential regulatory elements such as a promoter to enable expression of a target gene in an appropriate host cell.
  • Vectors known in the art can be used.
  • vectors may contain expression control elements that allow the coding region to be expressed correctly in a suitable host.
  • the recombinant cell or transformant refers to a cell transformed by introducing a vector having a polynucleotide encoding one or more target proteins into a host cell, and is a method for producing a transformant by introducing an expression vector into a host cell.
  • Examples include the calcium phosphate method or the calcium chloride/rubidium chloride method described in the literature (Sambrook, J., et al., Molecular Cloning, A Laboratory Manual (2nd edition), Cold Spring Harbor Laboratory, 1. 74, 1989), electrophoresis There are methods such as electroporation, electroinjection, chemical treatment methods such as PEG, and methods using gene guns.
  • antibody proteins By culturing the transformant expressing the vector in a nutrient medium, antibody proteins can be produced and isolated in large quantities.
  • the medium and culture conditions can be appropriately selected and used depending on the host cell tolerance. During culture, conditions such as temperature, pH of the medium, and culture time must be appropriately adjusted to suit cell growth and mass production of proteins.
  • the vector according to the present invention may be preferably a hepatocyte, more preferably a hepatocarcinoma-derived cell, but cells expressing miR-122 and known in the art can be applied without limitation.
  • the present invention relates to a method for selecting a target substance for miR-122 for inhibiting HCV proliferation using a dual-measurement luminescence platform for selecting a target substance for miR-122 for inhibiting HCV proliferation.
  • the selection method includes the steps of (a) treating a candidate candidate for the miR-122 target on a recombinant cell transformed with a dual-measurement luminescent platform for selecting a target material for miR-122 to inhibit HCV proliferation or a vector containing the same; and
  • the miR-122 binding site-Rluc (Renilla luciferase)-HCV replicon (HIRL-2Aneo-NSrep) and Fluc (Firefly luciferase)-miR-122 binding site A dual instrument luminescence platform composed of mRNA (FL-5 ⁇ 122) was fabricated. When the expression of miR-122 increases, the HCV replicon is replicated by miR-122, resulting in an increase in the Rluc signal and a decrease in the Fluc signal.
  • HCV replicon replication is reduced and the Rluc signal is decreased, while the Fluc signal is increased, so it can be applied to select a target substance for miR-122 to inhibit HCV proliferation.
  • the HCV/miR-122 activity dual measurement luminescent platform of the present invention can be effectively applied to select miR-122 target substances for inhibiting HCV proliferation.
  • the present invention provides a sofosbuvir-resistant HCV therapeutic agent consisting of a miR-122 binding portion, a reporter gene, and an HCV replicon, and represented by the RNA base sequence of SEQ ID NO: 3 or SEQ ID NO: 4. It is about a light-emitting platform for sorting.
  • the light-emitting platform for screening sofosbuvir-resistant HCV therapeutic substances is such that the amino acid encoded by the 844th and 846th base sequence of the NS5B region of the wild-type HCV replicon is substituted from serine to threonine (S282T). It is characterized by being designed.
  • the 7841st base in the RNA base sequence of SEQ ID NO: 3 is substituted with C, and the 7840th to 7842nd base sequence in the RNA base sequence of SEQ ID NO: 3 encodes threonine.
  • RNA base sequence of SEQ ID NO: 4 is substituted with C, and the 7725th to 7727th base sequence in the RNA base sequence of SEQ ID NO: 4 encodes threonine.
  • the reporter gene is characterized as being a Renilla luciferase gene or a Firefly luciferase gene.
  • a reporter protein gene known in the art may be used according to the present invention. Can be applied to.
  • the light-emitting platform is composed of a Firefly luciferase gene and a miR-122 binding site, and additionally includes a miR-122 measurement light-emitting platform represented by the base sequence of SEQ ID NO: 2. It can be included. If the miR-122 measurement luminescence platform is additionally included, the reporter genes of the luminescence platform for screening sofosbuvir-resistant HCV therapeutic substances and the miR-122 measurement luminescence platform can be changed to emit light in different colors or wavelengths.
  • the present invention in another aspect of the present invention, it relates to a vector containing a light-emitting platform for selecting a sofosbuvir-resistant HCV treatment substance. In another aspect, the present invention relates to a vector containing a light-emitting platform for selecting a sofosbuvir-resistant HCV treatment substance. .
  • the present invention relates to a light-emitting platform for selecting a sofosbuvir-resistant HCV therapeutic agent or a recombinant cell transformed with a vector containing the same.
  • the present invention relates to a method for selecting a sofosbuvir-resistant HCV therapeutic material using the luminescent platform for screening a sofosbuvir-resistant HCV therapeutic material.
  • the selection method includes the steps of (a) treating a sofosbuvir-resistant HCV treatment candidate to recombinant cells transformed with a light-emitting platform for selecting a sofosbuvir-resistant HCV treatment material or a vector containing the same; and
  • the present invention relates to a method of providing information to administer rigosertib to patients with hepatitis C virus infection who are resistant to sofosbuvir.
  • a light-emitting platform for screening sofosbuvir-resistant HCV therapeutic substances was manufactured as shown in Figures 4 and 5.
  • HCV subgenomic replicon geneotype 1b or genotype 2a
  • S282T substitution in the NS5B region was transfected into Huh7 cells and treated with rigosertib and sofosbuvir, respectively
  • Rigosert Tip was confirmed to inhibit proliferation of both wild-type HCV and mutant (S282T) HCV.
  • sofosbuvir inhibited the proliferation of wild-type HCV, but did not inhibit the proliferation of mutant (S282T) HCV.
  • Sofosbuvir is known to be effective in eliminating hepatitis C virus, but has the problem of being resistant to mutant viruses. That is, in the present invention, since it was confirmed that ligosertib effectively inhibits the proliferation of mutant HCV resistant to sofosbuvir, ligosertib can be used as a composition for preventing or treating sofosbuvir-resistant HCV.
  • the present invention in another aspect, relates to a pharmaceutical composition for preventing or treating sofosbuvir-resistant HCV, comprising rigosertib.
  • the present invention provides hepatitis C virus resistant to sofosbuvir, comprising the step of providing information on predicting the effect of administering rigosertib to patients with hepatitis C virus infection showing resistance to sofosbuvir. It is about how to provide information about treatment.
  • the pharmaceutical composition of the present invention can be formulated and used in various forms according to conventional methods.
  • it can be formulated into oral dosage forms such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, etc., and can be formulated and used in the form of external preparations, suppositories, and sterile injection solutions.
  • it may further include pharmaceutically acceptable carriers, excipients, and diluents.
  • it can be formulated and used in the form of external preparations such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, etc., and sterile injection solutions according to conventional methods.
  • the carriers, excipients and diluents include lactose, dextrose, sucrose, oligosaccharides, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum acacia, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, These include microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxy benzoate, propylhydroxy benzoate, talc, magnesium stearate, and mineral oil.
  • diluents or excipients such as commonly used fillers, extenders, binders, wetting agents, disintegrants, and surfactants.
  • Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc. These solid preparations contain at least one excipient, such as starch, calcium carbonate, or sucrose. It is prepared by mixing , lactose, and gelatin. In addition to simple excipients, lubricants such as magnesium styrate talc are also used.
  • Liquid preparations for oral use include suspensions, oral solutions, emulsions, and syrups. In addition to the commonly used simple diluents such as water and liquid paraffin, various excipients such as wetting agents, sweeteners, fragrances, and preservatives may be included. there is.
  • Preparations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, suppositories, etc.
  • Non-aqueous solvents and suspensions may include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, and injectable ester such as ethyl oleate.
  • injectable ester such as ethyl oleate.
  • As a base for suppositories witepsol, macrogol, tween 61, cacao, laurin, glycerogenatin, etc. can be used.
  • the term “administration” means providing the pharmaceutical composition of the present invention to an individual by any suitable method.
  • the present invention provides a pharmaceutical composition that provides an amount of an active ingredient or pharmaceutical composition that induces a biological or medical response in a tissue system, animal or human as considered by a researcher, veterinarian, doctor or other clinician, that is, alleviation of symptoms of the disease or disorder being treated. It can be administered in a therapeutically effective amount that induces . It is obvious to those skilled in the art that the therapeutically effective dosage and frequency of administration of the pharmaceutical composition of the present invention will vary depending on the desired effect.
  • the optimal dosage to be administered can be easily determined by a person skilled in the art, depending on the type of disease, the severity of the disease, the content of the active ingredient and other ingredients contained in the composition, the type of dosage form, the patient's age, weight, and general health condition. , gender and diet, administration time, administration route and secretion rate of the composition, treatment period, and various factors including concurrently used drugs.
  • the pharmaceutical composition of the present invention can be administered to an individual through various routes. For example, it may be administered intravenously, intraperitoneally, intramuscularly, intraarterially, bucally, intracardiacally, intramedullary, intrathecally, transdermally, enterally, subcutaneously, sublingually, or topically, but is not limited thereto.
  • the pharmaceutical composition of the present invention can be administered in an amount of 1 to 10,000 mg/kg/day, and may be administered once a day or divided into several times.
  • Example 1 Preparation of a dual-measurement luminescence platform for selection of miR-122 target substances for inhibiting HCV proliferation
  • HIRL-2Aneo-NSrep (NKR2AN) DNA was treated with Sca I restriction enzyme, and RNA was synthesized using the linearized DNA.
  • HIRL-2Aneo-NSrep RNA (SEQ ID NO: 1) synthesized by electroporation is introduced into the Huh7 cell line, and the HCV replicon can be measured with Renilla luciferase through G418 drug selection. A stable cell line was first prepared.
  • pLJ-FL-5 ⁇ 122 virus was generated with Firefly luciferase and the miR-122 sensor (SEQ ID NO. 2) of five consecutive miR-122 Seed sequences. and transfect. This was treated in combination with G418 (500 ⁇ g/ml) and blasticidin (5 ⁇ g/ml) to prepare a transformed cell line system with a dual instrument luminescence platform capable of selecting final HCV and miR-122 target substances.
  • Example 2 Selection of miR-122 target substances for inhibiting HCV proliferation
  • Ligosertip showed the strongest effect on HCV replicon and miR-sensor activity. It was confirmed that it was shown.
  • Huh7 cells transformed with the dual instrument luminescence platform were treated with ligosertip at various concentrations (0, 0.625, 1.25, 2.5, 5, and 10 ⁇ M) for 48 hours, and then the lysate of each cell was double-luciferated.
  • Renilla luciferase which measures HCV replicon
  • Firefly luciferase which measures miR-122
  • RNA was extracted using a known general method.
  • cDNA was synthesized from the extracted RNA through the reverse transcription process of the Mir- Quantitative real-time PCR (qRT-PCR) was performed using -5'-TGGAGTGTGACAATGGTGTTTG-3'; (Seo et al., Proceedings of the National Academy of Sciences U S A, 119(51), e2214911119, 2022).
  • Example 3 Preparation of a luminescent platform for screening sofosbuvir-resistant HCV therapeutic substances
  • HIRL-2Aneo-NSrep-5B (S282T), represented by the RNA base sequence of SEQ ID NO: 3, and HIRL-2Aneo-NSrep-5B (S282T), represented by the RNA base sequence of SEQ ID NO: 4, are luminescent platforms for selecting sofosbuvir-resistant HCV therapeutic substances.
  • HIFL-NSrep-5B(S282T) was prepared.
  • the S282T mutation, located in the NS5B region, is where Serine (S, Serine), the 282nd amino acid sequence, is replaced with Threonine (T), and shows resistance to the drug sofosbuvir.
  • the cell line system is treated with drugs such as rigosertib and sofosbuvir at different concentrations (0, 0.5, 1 ⁇ M) for 48 hours, and then luciferase activity analysis is performed.
  • drugs such as rigosertib and sofosbuvir at different concentrations (0, 0.5, 1 ⁇ M) for 48 hours, and then luciferase activity analysis is performed.
  • HIRL-2Aneo-NSrep-5B(S282T) was measured for Renilla luciferase
  • HIFL-NSrep-5B(S282T) was measured for Firefly luciferase activity and compared with the wild-type control.
  • the HCV/miR-122 activity dual measurement luminescent platform of the present invention can be effectively applied to select miR-122 target substances for inhibiting HCV proliferation, and the luminescent platform for screening sofosbuvir-resistant HCV therapeutic substances is a sofosbuvir-resistant HCV therapeutic agent. It can be easily applied for screening.
  • the ligosertib selected using the platform of the present invention shows strong antiviral activity against sofosbuvir-resistant HCV, it can be usefully used as a composition for preventing or treating sofosbuvir-resistant HCV.

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Abstract

La présente invention concerne une plateforme luminescente pour la double mesure de l'activité du VHC/miR-122 et l'utilisation d'une substance rigosertib dérivée pour le traitement contre le VHC résistant au sofosbuvir. Dans la plateforme luminescente selon la présente invention pour la double mesure de l'activité du VHC/miR-122 pour cribler des substances ciblant miR-122 pour l'inhibition de la prolifération du VHC, il a été confirmé que la réplication de réplicons du VHC diminue en présence d'une substance ciblant miR-122, ce qui conduit à une diminution du signal provenant de la plateforme luminescente pour la mesure de la prolifération du VHC et à une augmentation du signal provenant de la plateforme luminescente pour la mesure de miR-122 et ainsi, la plateforme luminescente selon la présente invention pour la double mesure de l'activité du VHC/miR-122 peut être efficacement appliquée au criblage de substances ciblant miR-122 pour l'inhibition de la prolifération du VHC. De plus, il a été confirmé que le rigosertib, qui a été dérivé à l'aide de la plateforme luminescente pour l'activité du VHC/miR-122 dans laquelle la région NS5B du réplicon du VHC a subi une substitution S282T, présente une puissante activité antivirale contre le virus de l'hépatite C portant une substitution d'acides aminés conférant de la résistance (VHC résistant au sofosbuvir) et ainsi, la plateforme selon la présente invention peut être facilement appliquée au criblage d'agents thérapeutiques contre le VHC résistant au sofosbuvir.
PCT/KR2023/019800 2022-12-05 2023-12-04 Plateforme luminescente pour la double mesure de l'activité du vhc/mir-122 et utilisation d'une substance rigosertib dérivée pour le traitement contre le vhc résistant au sofosbuvir Ceased WO2024123010A1 (fr)

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KR10-2022-0167321 2022-12-05
KR20220167321 2022-12-05
KR1020230173335A KR20240087568A (ko) 2022-12-05 2023-12-04 HCV/miR-122 활성 이중계측 발광플랫폼 및 도출물질 리고세르팁의 소포스부비어 저항성 HCV 치료 용도
KR10-2023-0173335 2023-12-04

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Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2011091209A1 (fr) * 2010-01-21 2011-07-28 North Carolina State University Modificateurs à petite molécule de microarn mir-122
WO2018172759A1 (fr) * 2017-03-20 2018-09-27 Queen Mary University Of London Procédé d'identification de patients résistants au sofosbuvir
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