WO2020262555A1 - 核酸医薬副作用軽減剤、該核酸医薬副作用軽減剤を含む医薬組成物、並びに核酸医薬の副作用惹起性を軽減する方法 - Google Patents

核酸医薬副作用軽減剤、該核酸医薬副作用軽減剤を含む医薬組成物、並びに核酸医薬の副作用惹起性を軽減する方法 Download PDF

Info

Publication number
WO2020262555A1
WO2020262555A1 PCT/JP2020/025093 JP2020025093W WO2020262555A1 WO 2020262555 A1 WO2020262555 A1 WO 2020262555A1 JP 2020025093 W JP2020025093 W JP 2020025093W WO 2020262555 A1 WO2020262555 A1 WO 2020262555A1
Authority
WO
WIPO (PCT)
Prior art keywords
nucleic acid
side effect
reducing agent
double
effect reducing
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
PCT/JP2020/025093
Other languages
English (en)
French (fr)
Japanese (ja)
Inventor
隆徳 横田
永田　哲也
倫太朗 原
正裕 大原
猛 和田
佐藤　一樹
雄介 前田
一憲 高木
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Tokyo University of Science
Tokyo Medical and Dental University NUC
Original Assignee
Tokyo University of Science
Tokyo Medical and Dental University NUC
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Tokyo University of Science, Tokyo Medical and Dental University NUC filed Critical Tokyo University of Science
Priority to JP2021527744A priority Critical patent/JPWO2020262555A1/ja
Publication of WO2020262555A1 publication Critical patent/WO2020262555A1/ja
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

Links

Images

Classifications

    • 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
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/7088—Compounds having three or more nucleosides or nucleotides
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/04—Peptides having up to 20 amino acids in a fully defined sequence; Derivatives thereof
    • A61K38/08—Peptides having 5 to 11 amino acids
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/04—Peptides having up to 20 amino acids in a fully defined sequence; Derivatives thereof
    • A61K38/10—Peptides having 12 to 20 amino acids
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
    • A61K47/42—Proteins; Polypeptides; Degradation products thereof; Derivatives thereof, e.g. albumin, gelatin or zein
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P39/00—General protective or antinoxious agents
    • A61P39/04—Chelating agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2/00—Peptides of undefined number of amino acids; Derivatives thereof
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
    • C12N15/113—Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing

Definitions

  • the present disclosure relates to a nucleic acid drug side effect reducing agent, a pharmaceutical composition containing the nucleic acid drug side effect reducing agent, and a method for reducing the side effect inducing property of the nucleic acid drug.
  • nucleic acid drugs that control the expression of specific genes or non-coding regions by utilizing the specificity of nucleic acid-nucleic acid interaction have been developed, and their clinical application has begun.
  • WO 2014/148620 describes a double-stranded nucleic acid binder, which controls the nuclease-degradability of nucleic acids.
  • the present disclosure relates to a nucleic acid drug side effect reducing agent that reduces side effects as a class effect of a nucleic acid drug, a pharmaceutical composition containing the nucleic acid drug side effect reducing agent, and a method for reducing the side effect inducement of the nucleic acid drug.
  • ⁇ 1> It is composed of an oligopeptide containing an oligopeptide region consisting of 2 to 40 amino acids including a structure W having a structure in which two or more amino acid residues of the following formula (I) are continuous.
  • a structure W having a structure in which two or more amino acid residues of the following formula (I) are continuous.
  • adjacent structures W are linked by one amino acid residue X not of formula (I).
  • the oligopeptide region is composed of one structure W or two or more structures W and an amino acid residue X.
  • Nucleic acid drug Side effect reducing agent [In formula (I), R 1 is the group H 3 N + -CH 2- or the group represented by formula (II).
  • R 2 does not exist when R 1 is a group H 3 N + -CH 2- , or is an alkylene group having 1 to 3 carbon atoms, and R 1 is a group represented by the formula (II). In the case, it is an alkylene group having 1 to 4 carbon atoms. R 1 and R 2 are all the same in one oligopeptide region. ] [In formula (II), R 3 , R 4 and R 5 are independently hydrogen atoms or methyl groups, respectively. ] ⁇ 2> R 1 is the nucleic acid drug side effect reducing agent according to ⁇ 1>, which is the basis of the formula (II).
  • nucleic acid drug side effect reducing agent according to ⁇ 2>, wherein R 3 , R 4 and R 5 of the group of the formula (II) are all hydrogen atoms.
  • R 3 , R 4 and R 5 of the group of the formula (II) are all hydrogen atoms.
  • R 1 is a group H 3 N + -CH 2- .
  • ⁇ 5> The nucleic acid drug side effect reducing agent according to any one of ⁇ 1> to ⁇ 4>, which is used for reducing side effects of a nucleic acid drug containing a single-stranded nucleic acid.
  • nucleic acid drug side effect reducing agent according to ⁇ 5>, wherein the single-stranded nucleic acid is a single-stranded RNA or a single-stranded DNA.
  • ⁇ 7> The nucleic acid drug side effect reducing agent according to any one of ⁇ 1> to ⁇ 4>, which is used for reducing side effects of a nucleic acid drug containing a nucleic acid containing a double-stranded nucleic acid structure.
  • nucleic acid drug side effect reducing agent according to ⁇ 7> or ⁇ 8>, wherein in the double-stranded nucleic acid structure, at least one nucleic acid strand satisfies at least one of the following conditions (i) and (ii): (I) The total number of ribonucleosides and unnatural nucleosides contained in the nucleic acid chain is 30% or more of the total number of nucleosides contained in the nucleic acid chain. (Ii) The total number of ribonucleosides and unnatural nucleosides contained in the nucleic acid chain is 6 That is all.
  • nucleic acid according to any one of ⁇ 7> to ⁇ 9>, wherein the nucleic acid is a single-stranded nucleic acid, and the double-stranded nucleic acid structure is formed by intramolecular hybridization of the single-stranded nucleic acid. Nucleic acid side effect reducing agent.
  • nucleic acid drug side effect reducing agent according to any one of ⁇ 7> to ⁇ 9>, wherein the double-stranded nucleic acid structure is formed by intermolecular hybridization.
  • the double-stranded nucleic acid structure includes an RNA-DNA complex double-stranded nucleic acid structure.
  • nucleic acid is a type A double-stranded nucleic acid.
  • the double-stranded nucleic acid structure contains a type A double-stranded nucleic acid structure, and the first nucleic acid strand of the complementary strands in the nucleic acid is a nucleic acid strand containing a structure in which four or more deoxyribonucleosides are continuously arranged.
  • the second nucleic acid strand of the complementary strands in the nucleic acid has a base sequence complementary to the first nucleic acid strand and contains at least one of RNA and PNA, according to ⁇ 11>.
  • the first nucleic acid chain of the complementary strand is a region containing an unnatural nucleoside continuously or discontinuously on the 5'-terminal side and / or 3'-terminal side of a region consisting of four or more consecutively linked deoxyribonucleosides.
  • the nucleic acid drug side effect reducing agent according to ⁇ 15> which is a complex DNA strand provided with.
  • the double-stranded nucleic acid structure contains a type A double-stranded nucleic acid structure
  • the first nucleic acid strand of the complementary strands in the nucleic acid is a structure in which segments composed of natural nucleosides and segments composed of unnatural nucleosides are alternately arranged.
  • a nucleic acid strand having, and having neither four or more consecutive deoxyribonucleotides nor four or more consecutive ribonucleotides, and the second nucleic acid strand of the complementary strands in the nucleic acid is the first nucleic acid strand.
  • the nucleic acid drug side effect reducing agent according to ⁇ 11> which has a base sequence complementary to the nucleic acid chain and contains at least one of RNA and PNA.
  • the second nucleic acid strand of the complementary strand is RNA, and the region complementary to the region of the first nucleic acid strand of the complementary strand containing the unnatural nucleoside is composed of the unnatural nucleoside.
  • nucleic acid drug side effect reducing agent according to any one of ⁇ 15> to ⁇ 19>, wherein the type A double-stranded nucleic acid structure includes an RNA-DNA complex double-stranded nucleic acid structure.
  • the unnatural nucleoside in the second nucleic acid strand of the complementary strand is a 2'-O-methylated and / or phosphorothioated nucleoside.
  • a functional molecule is bound to the second nucleic acid strand of the complementary strand.
  • nucleic acid drug side effect reducing agent according to ⁇ 22>, wherein the functional molecule is a molecule having an activity of delivering a nucleic acid containing the double-stranded nucleic acid structure to a target site.
  • the functional molecule is a molecule having an activity of delivering a nucleic acid containing the double-stranded nucleic acid structure to a target site.
  • R 2 of the formula (I) is a methylene group.
  • nucleic acid drug side effect reducing agent according to ⁇ 7>, wherein the double-stranded nucleic acid structure is a B-type double-stranded nucleic acid structure.
  • ⁇ 27> The nucleic acid drug side effect reducing agent according to ⁇ 26>, wherein the nucleic acid containing the double-stranded nucleic acid structure is double-stranded DNA.
  • R 2 of the formula (I) is a trimethylene group.
  • ⁇ 29> The nucleic acid drug side effect reducing agent according to any one of ⁇ 1> to ⁇ 28>, wherein all of the oligopeptide region comprises an amino acid residue of the formula (I).
  • ⁇ 30> One of ⁇ 1> to ⁇ 29>, which is used for reducing side effects of nucleic acid drugs containing nucleic acids having a length of 10 to 25 bases and has 6 to 34 amino acid residues in the oligopeptide region.
  • ⁇ 31> The nucleic acid drug side effect reducing agent according to ⁇ 30>, wherein the nucleic acid is siRNA as a double-stranded nucleic acid.
  • all the amino acid residues with optical activity are amino acid residues having L-type or all D-type optical activity, among ⁇ 1> to ⁇ 31>.
  • the nucleic acid drug side effect reducing agent according to any one.
  • ⁇ 33> The nucleic acid drug side effect reducing agent according to any one of ⁇ 1> to ⁇ 32>, wherein a delivery molecule having an activity of delivering nucleic acid to a target site is bound to the oligopeptide.
  • ⁇ 34> The nucleic acid drug side effect reducing agent according to any one of ⁇ 1> to ⁇ 33>, which promotes the decomposition of nucleic acid contained in a nucleic acid drug by RNase H.
  • ⁇ 35> Any one of ⁇ 1> to ⁇ 34> used at a ratio of 0.5 to 20 mol to 1 mol of the nucleic acid having a length of 10 to 50 base together with a nucleic acid drug containing a nucleic acid having a length of 10 to 50 base.
  • ⁇ 36> A pharmaceutical composition comprising the nucleic acid drug side effect reducing agent according to any one of ⁇ 1> to ⁇ 35> and the nucleic acid drug. ⁇ 37> 3.
  • nucleic acid drug according to ⁇ 36>, wherein the nucleic acid drug contains a nucleic acid having a length of 10 to 50 bases, and contains a nucleic acid drug side effect reducing agent at a ratio of 0.5 to 20 mol to 1 mol of the nucleic acid having a length of 10 to 50 bases.
  • Pharmaceutical composition. ⁇ 38> The nucleic acid drug contains an A-type double-stranded nucleic acid having a length of 15 to 25 bases, and the nucleic acid drug has a ratio of 0.8 to 2.5 mol to 1 mol of the A-type double-stranded nucleic acid having a length of 15 to 25 bases.
  • the pharmaceutical composition according to ⁇ 37> which comprises an antisense agent.
  • ⁇ 39> The pharmaceutical composition according to any one of ⁇ 36> to ⁇ 38>, which is used for treating or preventing a specific disease targeted by the nucleic acid drug while reducing side effects caused by the nucleic acid drug. .. ⁇ 40>
  • the pharmaceutical composition according to any one of ⁇ 36> to ⁇ 39> which is for rapid intravenous injection.
  • ⁇ 41> The pharmaceutical composition according to any one of ⁇ 36> to ⁇ 40>, wherein the single dose is 0.01 mg to 200 mg per kg of body weight of the subject to whom the pharmaceutical composition is administered, as the amount of nucleic acid. Stuff.
  • ⁇ 42> The pharmaceutical composition according to any one of ⁇ 36> to ⁇ 41>, further comprising at least one of polyethylene glycol (PEG) and glycerol.
  • PEG polyethylene glycol
  • ⁇ 43> A method for reducing the side effect-inducing property of a nucleic acid drug, which comprises adding the nucleic acid drug side effect reducing agent according to any one of ⁇ 1> to ⁇ 35> to the nucleic acid drug.
  • a nucleic acid drug side effect reducing agent for reducing side effects as a class effect of a nucleic acid drug a pharmaceutical composition containing the nucleic acid drug side effect reducing agent, and a method for reducing the side effect inducement of the nucleic acid drug are provided. Can be done.
  • the gene expression inhibitory effect of the nucleic acid drug when the nucleic acid drug is used together with the nucleic acid drug side effect reducing agent according to the present disclosure is shown.
  • the gene expression inhibitory effect of the nucleic acid drug when the nucleic acid drug is used together with the nucleic acid drug side effect reducing agent according to the present disclosure is shown.
  • the gene expression inhibitory effect of the nucleic acid drug when the nucleic acid drug is used together with the nucleic acid drug side effect reducing agent according to the present disclosure is shown. It is a measurement result of aPTT 30 minutes after administration of the cholesterol-binding nucleic acid complex to which Dab10 / Dab12 was added in Example 2. Error bars indicate standard error.
  • the numerical range indicated by using "-" in the present disclosure indicates a range including the numerical values before and after "-" as the minimum value and the maximum value, respectively.
  • the upper limit value or the lower limit value described in a certain numerical range may be replaced with the upper limit value or the lower limit value of another numerical range described stepwise.
  • the upper limit value or the lower limit value described in a certain numerical range may be replaced with the value shown in the examples.
  • the amount of each component in the composition is the total amount of the plurality of substances present in the composition unless otherwise specified, when a plurality of substances corresponding to each component are present in the composition. means.
  • each element may exist alone or in plurality.
  • the combination of preferred embodiments is a more preferred embodiment.
  • nucleic acid is used interchangeably with polynucleosides and oligonucleosides to refer to polymers or oligomers of nucleosides of any length.
  • Nucleic acid may be single-stranded or double-stranded unless otherwise specified.
  • the nucleoside linkage is not limited to the phosphodiester bond linkage found in the natural nucleic acid, and may be a phosphodiester bond or the like as described later. Therefore, in the present disclosure, expressions such as “continuous nucleoside”, “polynucleoside”, and “oligonucleoside” may be used without specifying the connection mode between nucleosides.
  • heteroduplex oligonucleoside may be referred to as “HDO (heteroduplex oligonucleoside)", and the antisense oligonucleoside may be referred to as “ASO (antisense oligonucleoside)”.
  • HDO heteroduplex oligonucleoside
  • ASO antisense oligonucleoside
  • the nucleic acid drug side effect reducing agent according to the present disclosure is a nucleic acid drug side effect reducing agent composed of an oligopeptide, and the oligopeptide has a structure W having a structure in which two or more amino acid residues of the following formula (I) are continuous.
  • a nucleic acid drug side effect reducing agent composed of an oligopeptide, and the oligopeptide has a structure W having a structure in which two or more amino acid residues of the following formula (I) are continuous.
  • an oligopeptide region consisting of 2 to 40 amino acids is contained and two or more structures W are present in the oligopeptide region, adjacent structures W are one amino acid residue not represented by the formula (I).
  • the oligopeptide region is composed of one structure W or two or more structures W and an amino acid residue X.
  • R 1 is the group H 3 N + -CH 2- or the group represented by formula (II).
  • R 2 does not exist when R 1 is a group H 3 N + -CH 2- , or is an alkylene group having 1 to 3 carbon atoms, and R 1 is a group represented by the formula (II). In the case, it is an alkylene group having 1 to 4 carbon atoms.
  • R 1 and R 2 are all the same in one oligopeptide region.
  • R 3 , R 4 and R 5 are independently hydrogen atoms or methyl groups, respectively.
  • the oligopeptides defined above are also referred to as oligopeptides according to the present disclosure.
  • R 3 , R 4 and R 5 may all be hydrogen atoms.
  • R 2 is an alkylene group having 1 to 3 carbon atoms
  • the alkylene group is, for example, a methylene group, an ethylene group, a 1,2-propylene group, or a trimethylene group.
  • R 2 is an alkylene group having 1 to 4 carbon atoms
  • the alkylene group is, for example, a methylene group, an ethylene group, a 1,2-propylene group, a trimethylene group, a 1,2-butylene group, 1,3-. It is a butylene group or a tetramethylene group.
  • n is the number of repetitions of the amino acid residue A. It is abbreviated as a positive integer indicating), and specific embodiments of a specific oligopeptide region are illustrated and listed.
  • a n corresponds to the structure W.
  • n is an integer of 2 to 40
  • n is preferably an integer of 4 to 20, more preferably an integer of 6 to 15.
  • specific oligopeptides region may be a structure represented by [A m -X] p.
  • Each A m corresponds to the above structure W.
  • each m is independently 2 or 3 or more integer
  • p is number of repetitions of [A m -X] units, A contained in the structure represented by [A m -X] p Is selected so that the number of is 2 to 40.
  • p is, for example, an integer of 2 to 20.
  • Each X independently represents an amino acid residue other than the amino acid residue of the formula (I). That [A m -X] structure represented by p, the amino acid residue 2 of formula (I), or between several stations comprising units of more than two, is an aspect that sandwich the other amino acid residues. Other amino acid residues can be inserted for the purpose of increasing or decreasing the degree of freedom (flexibility) of the specific oligopeptide region.
  • Specific examples of this aspect (2) include, but are not limited to, the following.
  • M preferably represents an integer of 2 to 20, and more preferably represents an integer of 2 to 10.
  • p preferably represents an integer of 2 to 20, and more preferably an integer of 2 to 10.
  • amino acid residues other than the amino acid residue represented by A are not particularly limited, and can be appropriately selected depending on the degree of freedom intended to be given to the oligopeptide.
  • Glycine can be mentioned as an amino acid residue having a large degree of freedom, but other amino acid residues that affect the degree of freedom of oligopeptides include, for example, L-alanine and L-proline, as well as amino acids having a proline skeleton. Examples thereof include, but are not limited to, L-aminoproline and L-guanidinoproline.
  • the portion of the oligopeptide according to the present disclosure other than the specific oligopeptide region may be appropriately selected as necessary, and is not particularly limited.
  • an amino acid derivative residue to which a signal source such as a protecting group or a fluorescent group such as fluorescein is bound may be used, if necessary.
  • So-called delivery molecules may be attached to the oligopeptides according to the present disclosure.
  • the delivery molecule include a molecule such as a signal peptide capable of introducing the oligopeptide according to the present disclosure into the cell, a molecule having target selectivity, and the like.
  • the oligopeptide according to the present disclosure can be selectively introduced into the liver or the like.
  • the lipid include cholesterol; fat-soluble vitamins such as vitamin E (tocopherols, tocotrienols, etc.), vitamin A, and vitamin K; intermediate metabolites such as acylcarnitine and acyl-CoA; glycolipids; lipids such as glycerides, and lipids. , Derivatives of these lipids.
  • cholesterol or vitamin E is given as a general preferable example from the viewpoint of safety and the like.
  • a sugar such as glucose or sucrose
  • binding the oligopeptide according to the present disclosure to various proteins and receptors on the cell surface of each organ is also recognized as a selective introduction means, antibodies and ligands specific to these cell surface proteins and the like are also recognized. Can also be used as a delivery molecule.
  • the oligopeptide according to the present disclosure may have its N-terminal protected by an acetyl group and / or its C-terminal protected by an amide group in order to bind the above-mentioned delivery molecule to the N-terminal or C-terminal.
  • an oligopeptide of the present disclosure oligonucleotides having the structure or [A m -X] amino acid residue group represented by Ac-YGG- the N-terminal side of the structure represented by p represented by A n It may be a peptide.
  • Ac represents an acetyl group.
  • amino acid residue is present at the N-terminal side of the structure represented by structural or [A m -X] p represented by A n, although the sequence of the N-terminal region is not particularly limited, its length It is preferably 1 to 5 amino acid residues. Amino acid residues may not be present in the N-terminal region. Similarly, if there is an amino acid residue at the C-terminal side of the structure represented by structural or [A m -X] p represented by A n, although the sequence of the C-terminal region is not particularly limited, its The length is preferably 1 to 5 amino acid residues. Amino acid residues may not be present in the C-terminal region.
  • Examples of the amino acid residue represented by A in the oligopeptide according to the present disclosure include the following structures.
  • the bars at the left and right ends of each structure represent the binding to adjacent amino acid residues.
  • the structural formula below shows the structure of amino acid residues derived from L-type amino acids
  • the amino acid residues represented by A may be amino acid residues derived from D-type amino acids.
  • the number of amino acid residues of the oligopeptide according to the present disclosure is not particularly limited, but may be, for example, 6 to 50 amino acid residues. Further, for example, in order to reduce the side effects of a nucleic acid drug containing a nucleic acid having a length of 10 to 25 bases, an oligopeptide having 6 to 34 amino acid residues in the oligopeptide region may be used, or an amino acid in the oligopeptide region may be used. An oligopeptide having 7 to 20 residues may be used, or an oligopeptide having 8 to 14 amino acid residues in the oligopeptide region may be used.
  • the oligopeptide according to the present disclosure can be produced according to a known chemical synthesis method for peptides. That is, it can be produced by using the liquid phase peptide synthesis method or the solid phase peptide synthesis method, which is now established as a conventional method.
  • a solid-phase peptide synthesis method generally recognized as a suitable chemical synthesis method, the Boc solid-phase method or the Fmoc solid-phase method can be used, and the ligation method can also be used if necessary. Is.
  • the required oligopeptides were synthesized using the Fmoc solid phase method, which is the most commonly used at present.
  • the individual amino acids constituting the oligopeptide can be produced by a known method, and a commercially available product can also be used.
  • the synthesized oligopeptide can be purified by a conventional method such as reverse phase high performance liquid chromatography (reverse phase HPLC) after a step such as deprotection according to a conventional method. Then, the target oligopeptide can be identified by mass spectrometry (matrix-assisted laser desorption ionization time-of-flight: MALDI-TOF or liquid chromatography-electrospray ionization: LC-ESI). Finally, the oligopeptide can be hydrolyzed to confirm the amino acid composition and content.
  • a conventional method such as reverse phase high performance liquid chromatography (reverse phase HPLC) after a step such as deprotection according to a conventional method. Then, the target oligopeptide can be identified by mass spectrometry (matrix-assisted laser desorption ionization time-of-flight: MALDI-TOF or liquid chromatography-electrospray ionization: LC-ESI). Finally
  • the chemical formula of the oligopeptide according to the present disclosure will be illustrated based on the state in which two amino acid residues, which are the smallest units constituting the specific oligopeptide region, are linked via a peptide bond.
  • the names given under each chemical formula are the names of the repeating structure after acetyltyrosine-glycine-glycine at the amino-terminal portion together with the number of repetitions, but they are also used as the names of the entire compound. ..
  • Dap is diaminopropionic acid
  • Dab is diaminobutanoic acid
  • Orn is ornithine
  • Lys is lysine
  • Agp is 2-amino-3-guanidino-propionic acid
  • Agb is 2-amino-3-guanidino-butanoic acid.
  • Arg represents arginine.
  • the oligopeptide according to the present disclosure it is one of the preferred embodiments of the oligopeptide according to the present disclosure that all the amino acid residues constituting the specific oligopeptide region are composed of the amino acid residues of the formula (I).
  • the "specific oligopeptide region” and other parts of the oligopeptide according to the present disclosure are derived from the above definition of "specific oligopeptide region”. That is, as long as two or more "amino acid residues other than formula (I)" are consecutive (however, as long as the condition "other than formula (I)” is satisfied, even if they are the same amino acid residues, they are different amino acids.
  • the location (which may be a residue) will be outside the "specific oligopeptide region".
  • the amino acid residue of the formula (I) closest to the place where two or more "amino acid residues other than the formula (I)" are continuous is ,
  • the C-terminal side and the N-terminal side are similarly "terminals of the specific oligopeptide region".
  • amino acid residues constituting the specific oligopeptide region it is preferable that all the amino acid residues with optical activity are L-type or all D-type amino acid residues having the same optical activity. ..
  • a so-called delivery molecule is bound to the oligopeptide according to the present disclosure. You may let me.
  • the oligopeptide according to the present disclosure it is possible to obtain an effect of suppressing the double-stranded nucleic acid or the double-stranded nucleic acid structure contained in the nucleic acid drug from being degraded by a nuclease other than RNase H.
  • a nucleic acid drug contains a nucleic acid having an RNA chain
  • the nucleic acid is rapidly degraded by RNase present in the blood or digestive tract, and the desired effect cannot be obtained. There is a tendency.
  • the oligopeptide according to the present disclosure has a function of protecting the nucleic acid from such degradation.
  • RNA-DNA complex double strands since the heteronucleic acid and the like described later function through the degradation of the sense strand by RNase H, it is not preferable that the degradation by RNase H is impaired.
  • RNase H specifically recognizes and degrades chimeric double strands such as RNA-DNA complex double strands.
  • the oligopeptide according to the present disclosure has a preferable property that it does not impair the degradability of RNA strands by RNase H.
  • nucleases other than RNase H that degrade the above-mentioned double-stranded nucleic acid or double-stranded nucleic acid structure include RNase A, oligonucleotidase, RNase II, RNase III, Spleen exonuclease, DNase I, DNase II, and so on. Examples thereof include Exodeoxyribonuclease VII. Although some nucleases existing in humans are listed here, nucleases existing in other organisms are also included in the above nuclease category. The range of the above nucleases includes both endonucleases and exonucleases.
  • RNase H is also referred to as Calf thymus ribonuclease H, Exolibonuclease H, or RNA-DNA-hybrid ribonucleotide hydrolase (hybrid ribonucleonucleotide double-stranded), and is an RNA-DNA complex double-stranded or the like. End-to-end degradation of strand RNA to produce 5'-phosphomonooligonucleotides or 5'-phosphonooligonucleotides.
  • RNase H is widely recognized from bacteria to mammals, and it is known that two or three types of RNase H are present in one cell.
  • the oligopeptide according to the present disclosure suppresses the degradation of nucleic acids contained in nucleic acid drugs by nucleases such as RNase A, but does not suppress the degradation of chimeric double strands by RNase H.
  • This seemingly contradictory function is based on the difference in nuclease binding sites in double-stranded nucleic acid or double-stranded nucleic acid structures.
  • RNase A is located on the major groove side of the double-stranded nucleic acid or double-stranded nucleic acid structure at the site where the oligopeptide according to the present disclosure binds. Since it is considered to bind, the action is inhibited by the presence of the oligopeptide according to the present disclosure. On the other hand, since RNase H binds to the minor groove side, it is considered that the degradation of RNA strand is not suppressed.
  • the nucleic acid drug in the present disclosure is not particularly limited as long as it is a nucleic acid drug that exerts a therapeutic or preventive effect on a disease or symptom when administered to a living body.
  • the nucleic acid contained in the nucleic acid drug may be a single-stranded nucleic acid or a double-stranded nucleic acid.
  • the nucleic acid drug may contain a single-stranded nucleic acid
  • the single-stranded nucleic acid may be a single-stranded RNA or a single-stranded DNA.
  • the double-stranded nucleic acid may be a type A double-stranded nucleic acid or a type B double-stranded nucleic acid.
  • the double-stranded nucleic acid may be double-stranded RNA, double-stranded DNA, or RNA-DNA complex double-stranded.
  • Double-stranded RNA and RNA-DNA complex double strands are known to have a type A double-stranded nucleic acid structure, at least in a physiological environment.
  • double-stranded DNA is known to have a B-type double-stranded nucleic acid structure, at least in a physiological environment.
  • the type A double-stranded nucleic acid may have a length of 15 to 25 bases.
  • the nucleic acid contained in the nucleic acid drug may be a nucleic acid containing a double-stranded nucleic acid structure.
  • the nucleic acid containing the double-stranded nucleic acid structure may entirely form a double-stranded nucleic acid, or only a part thereof may form a double-stranded nucleic acid.
  • Such a double-stranded nucleic acid structure is not limited to a structure formed by intermolecular hybridization of two nucleic acid molecules, and one nucleic acid molecule (single-stranded confirmation molecule) is self-complementary within the molecule.
  • the structure may be formed by intramolecular hybridization.
  • the description of double-stranded nucleic acids also applies to double-stranded nucleic acid structures as long as there is no contradiction.
  • the double-stranded nucleic acid structure may be an RNA-DNA complex double-stranded nucleic acid structure.
  • the double-stranded nucleic acid structure may include an A-type double-stranded nucleic acid structure or a B-type double-stranded nucleic acid structure.
  • the total length of the double-stranded nucleic acid structure may be an A-type double-stranded nucleic acid structure, or only a part of the double-stranded nucleic acid structure is an A-type double-stranded nucleic acid. It may be a structure. That is, it can be said that the double-stranded nucleic acid structure may at least partially include a type A double-stranded nucleic acid structure.
  • the total length of the double-stranded nucleic acid structure may be a B-type double-stranded nucleic acid structure, or only a part of the double-stranded nucleic acid structure is a B-type double-stranded nucleic acid. It may be a structure.
  • nucleic acid drugs include antisense nucleic acid, which is a single-stranded nucleic acid, siRNA (small interfering RNA), which is a double-stranded RNA, aptamer, which is a single-stranded nucleic acid, decoy, which is double-stranded DNA, and single-stranded DNA.
  • siRNA small interfering RNA
  • aptamer which is a single-stranded nucleic acid
  • decoy which is double-stranded DNA
  • single-stranded DNA and single-stranded DNA.
  • CpG oligo which is a double-stranded heteronucleic acid consisting of an RNA strand and a DNA strand, and the like.
  • At least one nucleic acid strand satisfies at least one of the following conditions (i) and (ii).
  • the total number of ribonucleosides and unnatural nucleosides contained in the nucleic acid chain is 30% or more of the total number of nucleosides contained in the nucleic acid chain.
  • the total number of ribonucleosides and unnatural nucleosides contained in the nucleic acid chain is 6 That is all.
  • the number of nucleosides contained in one nucleic acid strand of interest in the double-stranded nucleic acid structure is counted.
  • the total number of ribonucleosides and unnatural nucleosides contained in the nucleic acid chain is more preferably 40% or more, further preferably 50% or more of the total number of nucleosides contained in the nucleic acid chain. It is more preferably 60% or more.
  • the total number of ribonucleosides and unnatural nucleosides contained in the nucleic acid chain is more preferably 8 or more, further preferably 10 or more, and even more preferably 12 or more.
  • the antisense nucleic acid may be, for example, a nucleic acid that selectively binds to mRNA and interferes with gene expression by inhibiting mRNA degradation and / or splicing.
  • the antisense nucleic acid may be one that complementarily binds to miRNA (microRNA) to inhibit the action of miRNA and / or degrade miRNA.
  • the antisense nucleic acid may be DNA, RNA, or a chimeric oligonucleoside of DNA and RNA. In addition, it may be optionally chemically modified.
  • Examples of chemical modification are (i) S conversion of phosphate groups between nucleic acids, that is, conversion from phosphodiester bond to phosphorothioate bond, and (ii) modification of the hydroxy group at the 2-position of ribose, for example, methoxyethyl of O atom. Modification with a group or a methyl group, substitution of an OH group with an F atom, use of a crosslinked artificial nucleic acid such as (iii) 2'-4'-BNA (commonly known as LNA), AmNA, and the like. The structure of the bond between these modified nucleosides and adjacent nucleosides is shown below.
  • Antisense nucleic acids that target genes are typically 12 to 30 bases long. Antisense nucleic acids that target miRNAs are typically 12 to 16 bases long.
  • the sequence of the antisense nucleic acid does not have to be a sequence that is completely complementary to the mRNA or miRNA to be bound, but from the viewpoint of specificity, it is complementary to the mRNA or miRNA to be bound. The higher the value, the better.
  • a nucleic acid strand similar to the first nucleic acid strand in the specific double-stranded nucleic acid described later may be used as the antisense nucleic acid.
  • the siRNA is incorporated into RNA-induced silencing complex (RISC), which is a complex containing RNase, and is unwound, leaving only the antisense strand complementary to the RNA to be silenced, and the RNA to be silenced. Is decomposed by RISC.
  • RISC RNA-induced silencing complex
  • the siRNA may optionally undergo chemical modifications as described above.
  • a delivery molecule such as cell-penetrating peptide (CPP) may be bonded by a disulfide bond or the like.
  • CPP cell-penetrating peptide
  • the siRNA that targets a gene is basically 21 bases long. However, in the case of siRNA excised by the Dicer enzyme, each strand is basically 23 bases long because the 5'side of each strand is overhanged by 2 bases in length.
  • the sequence of the antisense strand does not have to be completely complementary to the RNA to be silenced, but from the viewpoint of specificity, the higher the complementarity, the
  • Aptamer is a single-stranded nucleic acid having a unique three-dimensional structure according to its sequence, and binds to a molecule such as a protein by the three-dimensional structure. By the binding, the activity of molecules such as proteins can be controlled, particularly the activity can be suppressed.
  • the antisense nucleic acid may be DNA, RNA, or a chimeric oligonucleoside of DNA and RNA. Aptamer lengths are typically 26-45 bases long.
  • Decoy is a double-stranded DNA that inhibits transcription of a specific genomic region by binding to a transcription factor.
  • the decoy sequence may be a sequence recognized by the target transcription factor, and the decoy length is typically about 20 bases long.
  • CpG oligo is a single-stranded DNA molecule that exerts a strong adjuvant effect, and activates the immune system by interacting with proteins such as Toll-like Receptor 9 (TLR9).
  • TLR9 Toll-like Receptor 9
  • the length of the CpG oligo is typically about 20 bases long.
  • Heteronucleic acid (hereinafter, also referred to as HDO) is a nucleic acid having a chimeric double-stranded portion such as RNA-DNA complex double-stranded portion.
  • the chimeric double strand is recognized by RNase H and the RNA strand is degraded. Therefore, by making the strand containing the DNA portion complementary to the RNA to be silenced, the RNA to be silenced can be degraded.
  • the heteronucleic acid may optionally undergo chemical modifications as described above. In addition, the delivery molecule may be bound.
  • the length of the heteronucleic acid is typically 12-45 bases long.
  • the sequence of the strand containing the DNA portion does not have to be completely complementary to the RNA to be silenced, but from the viewpoint of specificity, the higher the complementarity, the more preferable.
  • the nucleic acid contained in the nucleic acid drug may be a double-stranded nucleic acid (hereinafter, also referred to as a specific double-stranded nucleic acid) described below.
  • the specific double-stranded nucleic acid is a double-stranded nucleic acid in which a first nucleic acid strand and a second nucleic acid strand containing a complementary region which is a base sequence complementary to the first nucleic acid strand are bound, and is a first nucleic acid.
  • the chain contains at least one selected from the group consisting of natural nucleosides and unnatural nucleic acids.
  • the complementary region includes a region composed of RNA at least in part. Since the double-stranded nucleic acid region composed of the RNA strand and the DNA strand serves as a substrate for RNase H in the cell, an antisense effect in the cell can be obtained and the expression of the target gene can be suppressed.
  • the "antisense effect” means, for example, RNA editing such as splicing, RNA-protein binding, digestion by RNase H, etc. by forming a double strand of an antisense oligonucleoside and a transcript such as an RNA sense strand. It means that RNA translation such as RNA degradation and translation into protein is altered, and as a result, the level of biological activity of the protein, which is a translation of the transcript, or the transcript itself is reduced.
  • exon skipping can be caused by hybridization of an antisense oligonucleoside (eg, first nucleic acid strand) to a transcript.
  • an antisense oligonucleoside eg, first nucleic acid strand
  • degradation of the transcript may result from RNase H or the like recognizing a hybridization moiety between the antisense oligonucleoside and the transcript.
  • ASO antisense oligonucleoside
  • the ASO binds to the transcript (mRNA) of the target gene, forming a partial double strand. Will be done.
  • This double strand serves as a cover to prevent translation by the ribosome, thus inhibiting the expression of the protein encoded by the target gene.
  • a DNA-containing oligonucleoside is introduced into a cell as ASO, a partial DNA-RNA heteroduplex is formed.
  • This structure is recognized by RNase H, which results in the degradation of the mRNA of the target gene, thus inhibiting the expression of the protein encoded by the target gene, which is referred to as the RNase H-dependent pathway.
  • antisense effects can be provided by targeting introns of pre-mRNA. The antisense effect may also be brought about by targeting the miRNA, in which case the function of the miRNA may be inhibited and the expression of the gene for which the miRNA suppresses expression may be increased.
  • an “antisense oligonucleoside” or “antisense nucleic acid” comprises a (ie, complementary) base sequence capable of hybridizing to at least a transcript of a target gene or a target transcript, and is predominantly. It refers to a single-stranded oligonucleoside that can suppress the expression of a transcript of a target gene or the expression level of a target transcript by an antisense effect.
  • target gene or “target transcript” whose expression is suppressed, altered or modified by the antisense effect is not particularly limited.
  • the “target gene” include a gene derived from an organism into which the specific double-stranded nucleic acid according to the present disclosure is introduced, a gene whose expression is increased in various diseases, and the like.
  • the "transcription product of the target gene” is RNA transcribed from genomic DNA, such as mRNA, miRNA, and the like. For example, in the case of mRNA, it is RNA transcribed from genomic DNA encoding a protein.
  • the transcript may be unmodified RNA, unspliced RNA, and the like.
  • the "target transcript” may be not only mRNA but also non-coding RNA (ie, ncRNA) such as miRNA. Therefore, the “transcript” may be any RNA synthesized by DNA-dependent RNA polymerase. More generally, the “transcript” may be any RNA synthesized by DNA-dependent RNA polymerase.
  • the "target transcript” is, for example, Apolipoprotein B (ApoB) mRNA, scavenger receptor B1, SRB1 mRNA, metastasis-related lung adenocarcinoma transcript 1 ( metastasis associated lung adenocarcinoma transcript 1, MALAT1) non-coding RNA, microRNA-122 (miR-122), ⁇ -secretase 1 (beta-secretase 1, BACE1) mRNA, or PTEN (Phosphatase and Tensin Homolog Deleted from Chromosome 10) mRNA It may be.
  • Apolipoprotein B Apolipoprotein B
  • SRB1 SRB1
  • metastasis-related lung adenocarcinoma transcript 1 metastasis associated lung adenocarcinoma transcript 1, MALAT1
  • MALAT1 metastasis associated lung adenocarcinoma transcript 1
  • MALAT1 metastasis associated lung a
  • Nucleotide sequences of genes and transcripts can be obtained from known databases such as the NCBI (National Center for Biotechnology Information) database.
  • the nucleotide sequence of the microRNA is, for example, the miRBase database (Kozomara A, Griffiths-Jones S. NAR 2014 42: D68-D73; Kozomara A, Griffiths-Jones S. NAR 2011 39: D152-D157; Griffiths-Jones S, Sani. HK, van Dongen S, Enright AJ. NAR 2008 36: D154-D158; Griffiths-Jones S, Grocock RJ, van Dongen S, Bateman A, Enright AJ. NAR 2006 34: D140-D144; Griffiths-Jones It can be obtained from 32: D109-D111).
  • the bond containing a phosphorus atom is a phosphorothioate bond in at least one nucleic acid chain selected from the group consisting of the first nucleic acid chain and the second nucleic acid chain.
  • the phosphorothioate bond refers to a bond between nucleosides in which the non-crosslinked oxygen atom of the phosphodiester bond is replaced with a sulfur atom.
  • naturally nucleotide includes deoxyribonucleotides found in DNA and ribonucleotides found in RNA.
  • deoxyribonucleotide and ribonucleotide may also be referred to as “DNA nucleotide” and “RNA nucleotide”, respectively.
  • the "natural nucleoside” includes a deoxyribonucleoside contained in DNA and a ribonucleoside contained in RNA.
  • deoxyribonucleoside and “ribonucleoside” may also be referred to as “DNA nucleoside” and “RNA nucleoside”, respectively.
  • non-natural nucleotide refers to any nucleotide other than the natural nucleotide, and the “non-natural nucleotide” includes modified nucleotides and nucleotide mimetics.
  • non-natural nucleoside refers to any nucleoside other than natural nucleoside, and “unnatural nucleoside” includes modified nucleosides and nucleoside mimetics.
  • nucleoside mimic is a structure in which a nucleoside sugar or sugar and base, and, but not necessarily, a bond is replaced with another sugar, another sugar and base, or another bond.
  • nucleoside mimetic include a nucleoside mimetic having a cyclohexenyl, cyclohexyl, tetrahydropyranyl, bicyclic sugar mimic or tricyclic sugar mimic instead of the sugar of the nucleoside, and examples thereof include non-furanose sugars.
  • nucleoside mimics having units are examples.
  • nucleotide mimetic is a structure in which the nucleoside and / or binding of a nucleotide is replaced with a nucleoside mimetic and / or another binding.
  • Nucleic acid chains containing unnatural oligonucleosides have, for example, enhanced cell uptake, enhanced affinity for nucleic acid targets, increased stability or increased inhibitory activity in the presence of nucleases compared to nucleic acid chains containing natural oligonucleosides. It has characteristics such as.
  • nucleic acid chains containing unnatural oligonucleotides have, for example, enhanced cell uptake, enhanced affinity for nucleic acid targets, increased or inhibited stability in the presence of nucleases compared to nucleic acid chains containing natural oligonucleotides. It has properties such as increased activity.
  • modified nucleotide means a nucleotide having at least one of a modified sugar moiety, a modified nucleoside bond, and a modified nucleobase.
  • modified nucleoside means a nucleoside having at least one selected from the group consisting of a modified sugar moiety and a modified nucleobase.
  • modified nucleoside bond refers to a nucleoside bond that is different from the naturally occurring nucleoside bond (ie, phosphodiester bond). Modified nucleoside linkages are generally more nuclease-resistant bindings than naturally occurring nucleoside linkages.
  • the first nucleic acid chain contains at least one selected from the group consisting of natural nucleosides and unnatural nucleosides.
  • the first nucleic acid chain according to the present disclosure may contain both a natural nucleoside and an unnatural nucleoside.
  • the first nucleic acid strand includes two terminal regions containing 2 to 10 consecutive nucleosides from the 5'end and 3'end of the first nucleic acid strand, and a terminal region. It is preferably located in between and composed of a central region containing at least 4 nucleosides.
  • the nucleosides in the terminal region and the central region are not particularly limited and may contain at least one selected from the group consisting of natural nucleosides and unnatural nucleosides, and may contain both natural nucleosides and unnatural nucleosides. Good.
  • the nucleoside in the terminal region comprises at least one unnatural nucleoside and may contain both an unnatural nucleoside and a natural nucleoside.
  • the nucleoside in the central region is not particularly limited and may contain at least one selected from the group consisting of natural nucleosides and unnatural nucleosides, and may contain both natural nucleosides and unnatural nucleosides.
  • these regions may contain, for example, cross-linked nucleosides, nucleosides containing 2'-O-MOE groups, and the like.
  • the examples and preferable examples of the natural nucleoside and the unnatural nucleoside in the terminal region and the central region are the same as the examples and the preferable examples of the natural nucleoside and the unnatural nucleoside in the natural nucleoside and the unnatural nucleoside in the wing region described later.
  • Each of the two terminal regions in the first nucleic acid chain is preferably 2 to 10 bases in length, and more preferably contains 2 to 5 consecutive nucleosides.
  • the nucleoside contained in the terminal region of the first nucleic acid chain is not particularly limited, but when the terminal region contains an unnatural nucleoside, the region continuously containing the unnatural nucleoside may be referred to as a "wing region".
  • the central region of the first nucleic acid chain is preferably at least 4 bases long, and more preferably 4 to 12 bases long.
  • the nucleoside contained in the central region of the first nucleic acid chain is not particularly limited, but when the nucleoside contained in the central region is a natural nucleoside, a region containing four or more natural nucleosides in succession is referred to as a "gap region". In some cases.
  • the specific double-stranded nucleic acid according to the present disclosure may contain a nucleic acid structure that can be recognized by RNase H.
  • the nucleic acid structure that can be recognized by RNase H include a site that is cleaved by RNase H.
  • RNase H is not particularly limited as long as it is an enzyme capable of recognizing a specific double-stranded nucleic acid of an animal including human and cleaving RNA.
  • the specific double-stranded nucleic acid according to the present disclosure includes a structure in which four or more deoxyribonucleosides are continuously arranged in the first nucleic acid chain, and four or more consecutive ribonucleosides are arranged in the second nucleic acid chain described later.
  • the specific double-stranded nucleic acid is derived from a structure containing a complementary base pair of a structure in which four or more of the deoxyribonucleosides are continuously arranged and a structure in which four or more of the ribonucleosides are arranged in succession. It may be configured.
  • the specific double-stranded nucleic acid is a nucleic acid in which the first nucleic acid strand of the complementary strand has a structure in which four or more deoxyribonucleosides are continuously arranged, and the second nucleic acid strand of the complementary strand is said. It may be a nucleic acid containing a type A double-stranded nucleic acid structure, which has a base sequence complementary to the first nucleic acid strand and contains at least one of RNA and PNA.
  • the second nucleic acid strand may be composed of only one of RNA and PNA, and may further contain nucleic acid structures other than these.
  • the region of the base sequence complementary to the first nucleic acid strand in the second nucleic acid strand may be composed of at least one of RNA and PNA, and the nucleic acid structures other than these may be further added. It may be included.
  • the first nucleic acid chain of the complementary strand is provided with a region containing an unnatural nucleoside continuously or discontinuously on the 5'-terminal side and / or 3'-terminal side of a region composed of four or more consecutive deoxyribonucleosides. It may be a complex DNA strand.
  • the unnatural nucleoside in the first nucleic acid strand of the complementary strand may be an LNA nucleoside.
  • the first nucleic acid strand includes a gap region containing a structure in which four or more natural nucleosides are continuously arranged, and an unnatural nucleoside located on the 5'terminal side of the gap region.
  • the wing region on the 5'end side (5'wing region) and the wing region on the 3'end side (3'wing region) will be collectively referred to as a "wing region" unless otherwise specified.
  • the antisense effect can be obtained more effectively.
  • the first nucleic acid strand may be a "gapmer".
  • gap mer refers to a gap region (DNA gap region) containing a structure in which at least four deoxyribonucleosides are arranged consecutively, and non-located on the 5'end side and 3'end side of the gap region. It refers to a nucleic acid chain consisting of a region containing a natural nucleoside (5'wing region and 3'wing region).
  • the wing region preferably contains at least one of a structure in which unnatural nucleosides are continuously arranged from the 5'end of the gap region and a structure in which unnatural nucleosides are continuously arranged from the 3'end of the gap region.
  • the wing region on the 5'end side of the gap region may be referred to as a "5'wing region”
  • the wing region on the 3'end side of the gap region may be referred to as a "3'wing region”.
  • the base lengths (lengths) of the 5'wing region and the 3'wing region may be usually 2 to 10 base lengths, 2 to 7 base lengths, or 2 to 5 base lengths, respectively.
  • the 5'wing region and the 3'wing region may further contain a natural nucleoside as long as it contains a structure in which unnatural nucleosides are continuously arranged.
  • the unnatural nucleoside is preferably a sugar-modified nucleoside from the viewpoint of stability against a nuclease.
  • the "sugar-modified nucleoside” refers to a modified nucleoside containing a modified sugar.
  • the "modified sugar” is a substitution of an arbitrary structure in a natural sugar (that is, a sugar moiety found in DNA (2'-H) or RNA (2'-OH)) with another structure. , Indicates a sugar having any variation from natural sugar.
  • Sugar-modified nucleosides can impart enhanced stability to nucleases, increased binding affinities, or other changes in molecular biological properties to nucleic acid chains.
  • the sugar-modified nucleoside contains a chemically modified ribofuranose ring moiety.
  • chemically modified ribofuranose rings include, but are not limited to, bicyclic nucleic acids (crosslinked nucleic acids, BNAs) by the addition of substituents (including 5'or 2'substituents) and the cross-linking of nongeminal ring atoms. ), S, N (R), or C (R 1 ) (R 2 ) (R, R 1 and R 2 of the ribosyl ring oxygen atom are each independently hydrogen atom and carbon number 1 to carbon number. Substitutions with 12 alkyls (representing protective groups), and combinations thereof.
  • the sugar-modified nucleoside may contain a 2'-modified sugar.
  • the 2'-modified sugar may be a sugar containing a 2'-O-methyl group.
  • "2'-modified sugar” means a furanosyl sugar modified at the 2'position.
  • sugar-modified nucleosides include, but are not limited to, 5'-vinyl, 5'-methyl (R or S), 4'-S, 2'-F (2'-fluoro group), 2'-.
  • examples include nucleosides containing OCH 3 (2'-OMe group or 2'-O-methyl group) or 2'-O (CH 2 ) 2 OCH 3 (2'-O-MOE) substituents.
  • "2'-modified sugar” means a furanosyl sugar modified at the 2'position.
  • sugar-modified nucleosides include bicyclic nucleosides.
  • bicyclic nucleoside refers to a modified nucleoside containing a bicyclic sugar moiety.
  • Nucleic acids containing bicyclic sugar moieties are commonly referred to as bridged nucleic acids (BNAs).
  • BNAs bridged nucleic acids
  • a nucleoside containing a bicyclic sugar moiety may also be referred to as a "crosslinked nucleoside”.
  • the bicyclic sugar may be a sugar in which a carbon atom at the 2'position and a carbon atom at the 4'position are crosslinked by two or more atoms.
  • Examples of bicyclic sugars include publicly known and publicly available ones.
  • BNAs bicyclic sugar-containing nucleic acids
  • p, m and n represent integers 1 to 4, integers 0 to 2 and integers 1 to 3, respectively; or R 3 is a hydrogen atom, an alkyl group, an alkenyl group, a cycloalkyl group, Cross-linked by aryl group, aralkyl group, acyl group, sulfonyl group, and unit substituent (representing a fluorescent or chemically luminescent labeled molecule, a functional group having nucleic acid cleavage activity, an intracellular
  • R 1 and R 2 are typical. Although they are hydrogen atoms, they may be the same or different from each other, and further, a hydroxy group protecting group, an alkyl group, an alkenyl group, a cycloalkyl group, an aryl group, an aralkyl group, etc. for nucleic acid synthesis.
  • Amino group alkoxy group having 1 to 5 carbon atoms, alkylthio group having 1 to 5 carbon atoms, cyanoalkoxy group having 1 to 6 carbon atoms, or having 1 to 5 carbon atoms It may represent an amino group substituted with an alkyl group).
  • the crosslinked nucleic acid is not particularly limited.
  • Known and publicly used cross-linked nucleic acid is also known as, for example, methyleneoxy (4'-CH 2- O-2') BNA (LNA (Locked Nucleic Acid®), 2', 4'-BNA.
  • ⁇ -L-methyleneoxy (4'-CH 2 -O-2') BNA or ⁇ -D-methyleneoxy (4'-CH 2- O-2') BNA ethyleneoxy (4' -(CH 2 ) 2 -O-2') BNA (also known as ENA), ⁇ -D-thio (4'-CH 2 -S-2') BNA, Aminooxy (4'-CH 2' -ON (R 3 ) -2') BNA, Oxyamino (4'-CH 2 -N (R 3 ) -O-2') BNA (also known as 2', 4'-BNA NC ), 2', 4'-BNA coc , 3'-amino-2', 4'-BNA, 5'-methyl BNA, (4'-CH (CH 3 ) -O-2') BNA (also known as cEt BNA) (4'-CH (CH 2 OCH 3 ) -O-2') BNA (also known as cMOE BNA), Amid BNA,
  • LNA nucleoside a crosslinked nucleoside having a methyleneoxy (4'-CH 2- O-2') crosslink (bicyclic nucleoside) may be referred to as "LNA nucleoside”.
  • the modified sugar can be prepared by a known and publicly available method.
  • the nucleobase moiety (natural, modified, or a combination thereof) may be maintained for hybridization with the target nucleic acid.
  • the sugar-modified nucleoside preferably contains a crosslinked nucleoside, and more preferably contains an LNA nucleoside.
  • the crosslinked nucleoside may contain a modified nucleobase.
  • the "modified nucleobase” or “modified nucleobase” means any nucleobase other than adenine, cytosine, guanine, thymine, or uracil.
  • the "unmodified nucleobase” or “unmodified nucleobase” is the purine bases adenine (A) and guanine (G), and the pyrimidine bases thymine (T), cytosine (C), and It means uracil (U).
  • modified nucleobases include 5-methylcytosine, 5-fluorocytosine, 5-bromocytosine, 5-iodocytosine or N4-methylcytosine; 5-fluorouracil, 5-bromouracil or 5-iodouracil; 2-thiothymine. N6-methyladenine or 8-bromoadenine; and N2-methylguanine or 8-bromoguanine and the like, but are not limited thereto.
  • the gap region is located between the 3'wing region and the 5'wing region and contains four or more natural nucleosides in succession.
  • the gap region is not particularly limited as long as it contains four or more natural nucleosides in succession, and may contain an unnatural nucleoside.
  • the gap region may contain a nucleoside containing a 2'-O-MOE group. Good.
  • a specific example of the unnatural nucleoside is synonymous with the unnatural nucleoside in the wing region, and the preferable range is also the same.
  • the base length of the gap region is preferably 4 to 20 bases, more preferably 4 to 15 bases, and even more preferably 4 to 10 bases.
  • the natural nucleoside is preferably deoxyribonucleoside or ribonucleoside, and more preferably deoxyribonucleoside.
  • the base length of the first nucleic acid strand is preferably 8 to 30 bases, more preferably 8 to 20 bases, and further preferably 10 to 15 bases. ..
  • the first nucleic acid chain may further contain a peptide nucleic acid.
  • Peptide nucleic acid is one of the above-mentioned nucleotide mimetics.
  • Peptide Nucleic Acid (PNA) is a nucleotide mimetic having a main chain in which N- (2-aminoethyl) glycine is bound by an amide bond instead of sugar.
  • the first nucleic acid strand may be a "mixer".
  • the term "mixed mer” means a segment consisting of a natural nucleoside (meaning at least one selected from the group consisting of a deoxyribonucleoside and a ribonucleoside) and a segment consisting of an unnatural nucleoside, which are arranged alternately. It may be periodic or random, and refers to a nucleic acid chain that does not have four or more consecutive deoxyribonucleosides and four or more consecutive ribonucleosides.
  • a mixmer in which the unnatural nucleoside is a cross-linked nucleoside and the natural nucleoside is a deoxyribonucleoside may be referred to as a "BNA / DNA mixmer".
  • BNA / RNA mixmer Mixmers in which the unnatural nucleoside is a cross-linked nucleoside and the natural nucleoside is a ribonucleoside may be referred to as a "BNA / RNA mixmer".
  • the mixmer does not necessarily have to be restricted to contain only two types of nucleosides (the two types here refer to types such as BNA and DNA, not types such as G and C).
  • the mixmer may contain any number of nucleosides, each type of nucleoside may be a natural nucleoside, a modified nucleoside, or a nucleoside mimic.
  • a mixmer may have deoxyribonucleoside segments separated from each other by a crosslinked nucleoside (eg, LNA nucleoside), and each deoxyribonucleoside segment may be formed from one deoxyribonucleoside or two. It may be formed from a series of deoxyribonucleosides.
  • the crosslinked nucleoside may contain a modified nucleobase (eg, 5-methylcytosine).
  • the second nucleic acid strand has a complementary region which is a base sequence complementary to the first nucleic acid strand. Therefore, in the specific double-stranded nucleic acid, the first nucleic acid strand is annealed to a complementary region in the second nucleic acid strand.
  • the complementary region in the second nucleic acid chain may be a natural nucleoside, an unnatural nucleoside, or both.
  • the complementary region may extend over the entire length of the second nucleic acid strand, but when an overhang region described later is present, the complementary region corresponds to a part of the length of the second nucleic acid strand.
  • "having a base sequence complementary to the first nucleic acid chain” also includes the case where a sequence other than the base sequence complementary to the first nucleic acid chain is further present. ..
  • the second nucleic acid when the first nucleic acid strand is composed of the wing region and the gap region and the gap region contains a deoxyribonucleoside, the second nucleic acid
  • the complementary region in the chain preferably contains a ribonucleoside, more preferably contains a structure in which ribonucleosides are continuously arranged, and more preferably three or more consecutive ribonucleosides, particularly preferably four. It is preferable to include a structure in which 5 consecutive or 5 consecutive structures are arranged.
  • the complementary region of the second nucleic acid strand When the complementary region of the second nucleic acid strand has such a structure in which ribonucleosides are continuously arranged, it can form a double strand with the DNA gap region of the first nucleic acid strand.
  • This double strand is recognized by RNase H and can cause cleavage of the second nucleic acid strand by RNase H.
  • the complementary region in the second nucleic acid chain may not include a structure in which two or more ribonucleosides are continuously arranged.
  • a "consecutive" nucleoside is a contiguous sequence of different nucleosides (eg, nucleosides with different bases) as long as the provisions for the nucleoside are met, even if the same nucleoside is not necessarily contiguous. You may.
  • the specific double-stranded nucleic acid according to the present disclosure is a specific double-stranded nucleic acid in which a first nucleic acid strand and a second nucleic acid strand having a complementary region which is a base sequence complementary to the first nucleic acid strand are bound.
  • the first nucleic acid chain is a 5'side wing containing a gap region containing a structure in which four or more deoxyribonucleosides are continuously arranged and a structure in which crosslinked nucleosides are continuously arranged from the 5'end of the gap region. It may have at least one of the 3'side wing regions containing a structure in which crosslinked nucleosides are lined up continuously from the 3'end of the region and the gap region.
  • the second nucleic acid chain preferably contains ribonucleoside.
  • a region complementary to the region containing the unnatural nucleotide of the first nucleic acid strand may be composed of unnatural nucleotides.
  • the unnatural nucleotide in the second nucleic acid chain may be an unnatural nucleotide having 2'-O-methylation and / or phosphorothioation.
  • the second nucleic acid chain may further contain at least one functional molecule bound to the polynucleoside.
  • the functional molecule may be linked to the 5'end of the second nucleic acid chain, may be linked to the 3'end, or may be linked to the nucleoside inside the polynucleoside.
  • the expressions "bonded to a nucleoside” and “bonded to a nucleoside” include cases where the phosphate group bound to the nucleoside, a phosphorothioate structure, and the like are bound.
  • the number of functional molecules in the second nucleic acid chain is not particularly limited and may be two or more.
  • the arrangement of the two or more functional molecules is not particularly limited, and the two or more functional molecules are linked to different positions of the polynucleoside. It may also be linked as a group to one position of the polynucleoside.
  • the bond between the second nucleic acid chain and the functional molecule may be directly linked or may be indirectly bound by another substance.
  • the functional molecule is preferably directly bound to the second nucleic acid chain via a covalent bond, an ionic bond, a hydrogen bond, or the like, from the viewpoint of obtaining a more stable bond. , It is more preferable that it is directly bound to the second nucleic acid chain via a covalent bond.
  • the functional molecule may be bound to the second nucleic acid chain via a cleavable linker moiety (linking group), and for example, the functional molecule may be linked via a disulfide bond.
  • linking group cleavable linker moiety
  • the functional molecule is selected from the group consisting of a specific double-stranded nucleic acid and a second nucleic acid strand to which the functional molecule is bound, and has one of a labeling function, a purification function, and a target delivery function.
  • the structure of the functional molecule is not particularly limited as long as it is a portion that imparts one or more.
  • the functional molecule preferably has at least one function selected from the group consisting of a labeling function, a purification function, and a target delivery function.
  • Examples of the portion that gives the labeling function include compounds such as fluorescent protein and luciferase.
  • Examples of the portion that provides the purification function include compounds such as biotin, avidin, His tag peptide, GST tag peptide, and FLAG tag peptide.
  • the functional molecule serves to enhance transport to the cell or cell nucleus.
  • certain peptide tags have been shown to enhance cellular uptake of oligonucleosides when conjugated to oligonucleosides. Examples include HaiFang Yin et al., Human Molecular Genetics, Vol. 17 (24), 3909-3918 (2008) and the arginine-rich peptides P007 and B peptides disclosed in their references.
  • Nuclear transport is enhanced by conjugating parts such as m3G-CAP (see Pedro M. D. Moreno et al., Nucleic Acids Res., Vol. 37, 1925-1935 (2009)) to oligonucleosides. be able to.
  • the specific double-stranded nucleic acid (or first nucleic acid strand) according to the present disclosure is delivered to a target site or target region in the body with high specificity and high efficiency, whereby a target transcript (for example, a target gene) by the related nucleic acid is delivered.
  • a target transcript for example, a target gene
  • a molecule having an activity of delivering the specific double-stranded nucleic acid of one embodiment of the present disclosure to a "target site" in the body becomes a second nucleic acid strand as a functional molecule. It is preferable that they are combined.
  • the specific double-stranded nucleic acid according to the present disclosure can be delivered to, for example, the liver with high specificity and high efficiency, and thus lipids, antibodies, peptides and proteins. It is preferably at least one molecule selected from.
  • lipids examples include lipids such as cholesterol and fatty acids (eg, vitamin E (tocopherols, tocotrienols), vitamin A, and vitamin D); fat-soluble vitamins such as vitamin K (eg, acylcarnitine); acyl-CoA and the like. Intermediate metabolites; glycolipids, glycerides, and derivatives thereof.
  • the lipid is preferably at least one selected from cholesterol, tocopherol, and tocotrienol.
  • the functional molecule is cholesterol or an analog thereof, tocopherol or an analog thereof, and a sugar (for example,). It may be glucose and sucrose).
  • the second nucleic acid chain may further include an overhang region located at at least one end selected from the group consisting of the 5'end and the 3'end of the complementary region.
  • the overhang region is preferably a single chain region.
  • the term "overhang region” means that when the first nucleic acid strand and the second nucleic acid strand are annealed to form a double-stranded nucleic acid structure, the 5'end of the second nucleic acid strand is 3 of the first nucleic acid strand. It consists of a nucleoside region in the second nucleic acid chain extending beyond the end and a nucleoside region in the second nucleic acid chain in which the 3'end of the second nucleic acid chain extends beyond the 5'end of the first nucleic acid chain. Indicates at least one region selected from the group. That is, the overhang region is a nucleoside region in the second nucleic acid strand that protrudes from the double-stranded nucleic acid structure and is a region adjacent to the complementary region.
  • the position of the overhang region is not particularly limited and may be present only on the 5'end side of the complementary region or only on the 3'end side of the complementary region. It may be present on both the 5'end side and the 3'end side.
  • the base length of each overhang region is 1 base length or more, preferably 9 base length or more.
  • the base length of each overhang region is, for example, 1 to 30 base lengths, preferably 9 to 17 base lengths, and more preferably 11 to 15 base lengths.
  • the lengths of the overhang regions may be the same as or different from each other.
  • the base length of the second nucleic acid chain is not particularly limited, but from the viewpoint of synthesis cost and delivery efficiency, it is preferably 40 base lengths or less, more preferably 18 to 30 base lengths, and further preferably 21 to 28 bases. It is long.
  • the base length of the second nucleic acid strand means the total base length of the complementary region and all the overhang regions.
  • the overhang region may be formed from a natural nucleoside, a non-natural nucleoside, or both a natural nucleoside and a non-natural nucleoside.
  • the overhang region in the second nucleic acid chain is preferably not a therapeutic oligonucleoside region.
  • the therapeutic oligonucleoside region is a region that exerts a function as, for example, an antisense oligonucleoside, a microRNA inhibitor (antimiR), a splice switching oligonucleoside, siRNA, a microRNA, or a pre-microRNA. Since the overhang region in the second nucleic acid chain does not have the therapeutic oligonucleoside region as described above, it does not have the ability to substantially hybridize to the intracellular transcript and does not easily affect gene expression. ..
  • At least one nucleoside (specifically, for example, 1 to 1 to 1) is present from the end.
  • 3 Nucleoside is preferably a sugar-modified nucleoside.
  • at least one nucleoside eg, at least two nucleosides or at least three nucleosides, more specifically 1-3 nucleosides from that end is a modified nucleoside. May be good.
  • the overhang region may be a region containing a sugar-modified nucleoside and having a base length of 9 to 12 bases.
  • the overhang region may be a region that does not contain a sugar-modified nucleoside and has a base length of 9 to 17 bases in the overhang region.
  • the first nucleic acid chain and the second nucleic acid chain each design the base sequence of the nucleic acid based on the information of, for example, the base sequence of the target transcript (or, in some cases, the base sequence of the target gene). Then, the nucleic acid chain is synthesized by using a commercially available automatic nucleic acid synthesizer (such as a product of Applied Biosystems, Inc., a product of Beckman Coulter, Inc.), and then a nucleic acid chain is synthesized. , The resulting oligonucleoside can be produced by purifying it using a reverse phase column or the like. Nucleic acid chains produced by this method are mixed in a suitable buffer solution, denatured at about 90 ° C.
  • the nucleic acids are denatured at about 30 ° C. to 70 ° C. for about 1 hour. It can be annealed for 8 hours to produce the specific double-stranded nucleic acid according to the present disclosure.
  • the production of specific double-stranded nucleic acids is not limited to the above time and temperature protocols. Suitable conditions for promoting double-stranded annealing are well known in the art.
  • the nucleic acid complex to which the functional molecule is bound can be produced by carrying out the above synthesis, purification and annealing using a nucleoside to which the functional molecule is bound in advance.
  • the method for linking the functional molecule to the nucleoside may be a known and publicly available method.
  • the nucleic acid strand constituting the specific double-stranded nucleic acid may be synthesized by designating the base sequence and the modification site or type.
  • the specific double-stranded nucleic acid according to the present disclosure is efficiently delivered in vivo, and the expression of the target gene or the level of the target transcript can be suppressed by the antisense effect. Therefore, the specific double-stranded nucleic acid according to the present disclosure may be used for expressing a target gene or suppressing the level of a target transcript.
  • nucleic acid base portion in the nucleic acid contained in the nucleic acid drug is shown below.
  • R T represents a hydrogen atom, an alkyl group, an alkenyl group, or alkynyl group
  • R C, R A and R G represents a hydrogen atom
  • R C2 represents a hydrogen atom or It represents an alkyl group
  • the wavy line represents a bond site with another structure.
  • the bond linking between nucleosides in a nucleic acid drug is not particularly limited, and examples thereof include a phosphorothioate bond, a phosphotriester bond, a methylphosphonate bond, a methylthiophosphonate bond, a boranophosphate bond, and a phosphoroamidate bond.
  • At least one nucleic acid chain selected from the group consisting of the first nucleic acid chain and the second nucleic acid chain includes a phosphorothioate bond as a bond linking the nucleosides.
  • the bond between deoxyribonucleosides is preferably a phosphorothioate bond.
  • the nucleic acid drug side effect reducing agent according to the present disclosure has at least one side effect as a class effect of the nucleic acid drug, for example, increase in APTT, thrombocytopenia, cerebral infarction, sedation. Reduce at least one effect, etc. Such an effect is the first effect found in the present disclosure. Surprisingly, even with the nucleic acid drug side effect reducing agent according to the present disclosure, the effectiveness of the nucleic acid drug is still in the effective range.
  • the nucleic acid drug side effect reducing agent according to the present disclosure does not reduce the side effect by simply preventing the interaction with other molecules of the nucleic acid drug, but the nucleic acid drug to the extent that the nucleic acid drug can exert its efficacy. It is considered that while allowing the interaction between the molecule and the target molecule, it suppresses the interaction with other molecules that cause side effects.
  • the amount of the oligopeptide according to the present disclosure is not particularly limited, but from the viewpoint of sufficiently exerting the effect of the nucleic acid drug while reducing the side effects of the nucleic acid drug (that is, treating or preventing a specific disease targeted by the nucleic acid drug).
  • a nucleic acid drug contains a nucleic acid having a length of 10 to 50 bases, it is preferable to use it at a ratio of 0.5 to 20.0 mol of the oligopeptide according to the present disclosure to 1 mol of the nucleic acid, preferably 0.6. It is more preferable to use it in a ratio of ⁇ 10.0 mol.
  • the oligopeptide according to the present disclosure may be used at a ratio of 0.8 to 5.0 mol or a ratio of 1.0 to 3.0 mol with respect to 1 mol of the nucleic acid. It may be used in a ratio of .2 to 2.7 mol, or may be used in a ratio of 1.5 to 2.5 mol. More specifically, when the nucleic acid drug contains a type A double-stranded nucleic acid structure-containing nucleic acid having a length of 15 to 25 bases, 0.5 to 10.0 for 1 mol of the type A double-stranded nucleic acid structure-containing nucleic acid.
  • the oligopeptide according to the present disclosure is 1.2 to 1.2 to 1 mol of the A-type double-stranded nucleic acid structure-containing nucleic acid. It may be used at a ratio of 1.7 mol, or may be used at a ratio of 1.5 to 2.5 mol.
  • the oligopeptide according to the present disclosure preferably contains 6 or more amino acid residues of the formula (I), and contains, for example, 5 amino acid residues of the formula (I). It may contain up to 20, 6 to 15, 8 to 14, or 10 to 12. Examples of these include Dab8, Dab10, Dab12, etc., which will be described later.
  • the present invention is based on 1 mol of the nucleic acid.
  • the amount (number of moles) of the oligopeptide according to the disclosure is preferably 0.4 ⁇ (T / S) or more, and more preferably 0.6 ⁇ (T / S) or more.
  • the amount is preferably 0.8 ⁇ (T / S) or more, and more preferably.
  • the amount (number of moles) of the oligopeptide according to the present disclosure may be 0.2 ⁇ (T / S) to 10.0 ⁇ (T / S) with respect to 1 mol of the nucleic acid, and 0.3 ⁇ (T). / S) to 5.0 ⁇ (T / S), 0.4 ⁇ (T / S) to 3.0 ⁇ (T / S), 0.4 ⁇ (T / S) to It may be 2.0 ⁇ (T / S), 0.4 ⁇ (T / S) to 1.5 ⁇ (T / S), and 0.6 ⁇ (T / S) to 1.2 ⁇ . It may be (T / S).
  • the base length of the nucleic acid means a length that does not include the overhang portion.
  • the oligopeptide When the oligopeptide according to the present disclosure is complexed with a nucleic acid in a nucleic acid drug, the oligopeptide may coexist with the nucleic acid in a buffer solution.
  • a buffer solution Although it is possible to use a phosphate buffer solution, it is possible to use a buffer solution that does not contain divalent anions, particularly phosphate ions, in consideration of the interaction with the phosphate group of the double-stranded nucleic acid. It is suitable for avoiding the aggregation phenomenon. Examples of suitable buffers include Tris buffer, HEPES buffer, and cacodylic acid buffer.
  • a solubility improver for improving the solubility of the oligopeptide and the nucleic acid drug in a solvent may be further used.
  • the solubility improver By using the solubility improver, the amount of oligopeptide and nucleic acid drug dissolved in a solution such as an injection solution can be increased, and the efficacy of the nucleic acid drug tends to be exhibited more efficiently.
  • the solubility improver include polyethylene glycol (PEG) and glycerol, and a more specific example thereof is PEG200.
  • the amount of the solubility improver used may be appropriately set according to the amount of the oligopeptide and the amount of the nucleic acid drug. For example, 1% by mass with respect to the total amount of the solution such as the injection solution containing the oligopeptide and the nucleic acid drug. It may be up to 50% by mass, or 5% by mass to 20% by mass.
  • the present disclosure also provides a pharmaceutical composition
  • a pharmaceutical composition comprising a nucleic acid drug side effect reducing agent and a nucleic acid drug according to the present disclosure.
  • the pharmaceutical composition can exert the efficacy of the nucleic acid drug while suppressing the side effect as a class effect of the nucleic acid drug.
  • the pharmaceutical composition according to the present disclosure may optionally further comprise a pharmaceutically acceptable carrier.
  • at least one of the above-mentioned solubility improvers, for example, PEG and glycerol, may be further contained.
  • the amount of the solubility improver used may be appropriately set according to the amount of oligopeptide and the amount of nucleic acid drug, but may be, for example, 1% by mass to 50% by mass with respect to the total amount of the pharmaceutical composition, and 5% by mass. It may be% to 20% by mass.
  • nucleic acid drug side effect reducing agent By using the nucleic acid drug side effect reducing agent according to the present disclosure, the side effects of the nucleic acid drug can be reduced. This gives you more freedom in choosing the dosage and administration of nucleic acid medications.
  • nucleic acid drugs cause the side effects described above as their class effects, so the dose must be kept low, or intramuscular injection, subcutaneous injection, infusion, etc. are used to suppress the increase in blood concentration.
  • the administration method had to be adopted. In other words, it was not possible to adopt a method of administration such as a large dose of rapid intravenous infusion. As a result, administration was not convenient for patients, and problems such as decreased adherence could occur.
  • the side effects of the nucleic acid drug can be reduced, so that a large dose can be administered (that is, the dose restriction is relaxed), and the dose restriction is relaxed. Since it is possible to adopt rapid intravenous injection, usage restrictions are relaxed. This has made it possible for patients to administer a required amount of nucleic acid drug in a convenient manner.
  • compositions can be formulated by a known pharmaceutical method.
  • pharmaceutical compositions include capsules, tablets, pills, liquids, powders, granules, fine granules, film coatings, pellets, troches, sublinguals, peptizers, buccal agents, pastes. , Syrups, Suspensions, Elixirs, Emulsions, Coatings, Ointments, Plasters, Pap (cataplasm), Transdermals, Lotions, Inhalants, Aerosols, Eye Drops, Injections and Suppositories It can be used orally or parenterally in the form of a drug.
  • pharmaceutically acceptable carriers or carriers that are acceptable as food and beverage products, specifically sterile water, physiological saline, vegetable oils, solvents, bases, emulsifiers, suspensions.
  • Agents, surfactants, pH regulators, stabilizers, flavors, fragrances, excipients, vehicles, preservatives, binders, diluents, isotonic agents, sedatives, bulking agents, disintegrants, buffers Agents, coatings, lubricants, colorants, sweeteners, thickeners, flavoring agents, solubilizers, and other additives can be incorporated appropriately.
  • the administration method of the pharmaceutical composition according to the present disclosure is not particularly limited, and for example, oral administration or parenteral administration, more specifically, intravenous administration, intraventricular administration, intrathecal administration, subcutaneous administration, intraarterial administration. , Intraperitoneal administration, intradermal administration, intrabronchial administration, rectal administration, intraocular administration, nasal administration and intramuscular administration, and administration by blood transfusion. From the viewpoint that the effect of the nucleic acid drug side effect reducing agent according to the present disclosure is remarkably exhibited, intravenous administration is preferable, and rapid intravenous injection is more preferable.
  • the specific double-stranded nucleic acid according to the present disclosure may not be bound to lipids such as vitamin E (tocopherol, tocotrienol) and cholesterol.
  • the use or method of the pharmaceutical composition according to the present disclosure is not particularly limited, and may be, for example, a use or method of administering to a cell to modify the function of the intracellular transcript, and the use or method of the intracellular protein. It may be a use or method that changes the expression level, or it may be a use or method that changes the protein structure in the cell.
  • the type of cell to which the pharmaceutical composition according to the present disclosure is administered is not particularly limited.
  • Examples of cell types include immune cells, epithelial cells, vascular endothelial cells, mesenchymal cells and the like.
  • the pharmaceutical composition according to the present disclosure can be used for animals including humans as a subject.
  • the animals other than humans are not particularly limited, and various livestock, poultry, pets, laboratory animals and the like can be subjects of some embodiments.
  • the dose or ingestion is appropriate according to the age, weight, symptom and health condition of the subject, the type of composition (pharmaceuticals, foods, beverages, etc.), etc. Can be selected for.
  • a large dose of the nucleic acid drug can be administered by the nucleic acid drug side effect reducing agent according to the present disclosure. Therefore, a single dose of the pharmaceutical composition according to the present disclosure (per 1 kg of body weight of the subject to be administered) is, for example, 0.01 mg / kg to 200 mg / kg, 0.1 mg / kg to the amount of nucleic acid.
  • the route of administration is as described above, and for example, intravenous injection can be adopted.
  • the administration frequency is, for example, once a day to once every four weeks, for example, once a week.
  • the amount of the oligopeptide according to the present disclosure in the pharmaceutical composition according to the present disclosure is the same as the amount of the oligopeptide according to the present disclosure in the description of the nucleic acid drug side effect reducing agent according to the present disclosure.
  • compositions according to the present disclosure are also used to treat or prevent diseases associated with, for example, gene mutations or increased expression of target genes (eg, metabolic diseases, tumors, and infectious diseases). May be good.
  • diseases associated with, for example, gene mutations or increased expression of target genes eg, metabolic diseases, tumors, and infectious diseases. May be good.
  • the pharmaceutical composition according to the present disclosure may be a pharmaceutical composition for administration in the ventricles or intrathecal space to treat or prevent a central nervous system disease.
  • the specific double-stranded nucleic acid used for intracerebroventricular or intrathecal administration may be one that does not bind lipids such as vitamin E (tocopherol, tocotrienol) and cholesterol.
  • the pharmaceutical composition according to the present disclosure may be administered to cells to treat a central nervous system disease.
  • central nervous system diseases include, but are not limited to, Huntington's disease, Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis (ALS), and brain tumors.
  • the pharmaceutical composition according to the present disclosure can be used as a therapeutic agent for cancer and various diseases, and as an antiviral agent for HIV and HCV.
  • a therapeutic agent for cancer and various diseases and as an antiviral agent for HIV and HCV.
  • renal cell cancer therapeutic agents targeting the FAK gene for example, renal cell cancer therapeutic agents targeting the FAK gene, age-related yellow spot degeneration (AMD) targeting the VEGF gene and therapeutic agents for yellow spot edema due to diabetes, VEGFR1 gene and RTP801 gene.
  • AMD age-related yellow spot degeneration
  • Age-related yellow spot degeneration (AMD) therapeutic agent targeted as a target gene pancreatic cancer therapeutic agent targeting protein kinase N ⁇ , liver cancer therapeutic agent targeting VEGF gene and kinesin spindle protein, ribonucleotide reductase M2 Liver cancer therapeutic agents targeting subsystems, hepatitis B therapeutic agents targeting the HBV gene, hepatitis C therapeutic agents targeting the HCV gene, RSV therapeutic agents targeting the RSV genome, influenza virus genome Targeted influenza drug, Th2 cytokine-targeted asthma drug, PCSK9 gene-targeted hypercholesterolemia drug, HIV-1 genome-targeted AIDS drug, hair growth gene-targeted hair loss Therapeutic agent, Parkinson's disease therapeutic agent targeting Huntingtin or ⁇ -synuclein, muscle atrophic lateral sclerosis (ALS) therapeutic agent targeting superoxide mustase, inflammatory disease therapeutic agent targeting TNF- ⁇ , Etc. can be mentioned.
  • ALD Age-related yellow spot degeneration
  • a method for treating or preventing a disease which comprises administering a therapeutically effective amount or a prophylactically effective amount of the pharmaceutical composition according to the present disclosure to a subject requiring administration of the pharmaceutical composition according to the present disclosure.
  • this method it is possible to select an appropriate dose and usage for treatment with a high degree of freedom while reducing side effects as a class effect of nucleic acid drugs.
  • a method for reducing the side effect-inducing property of a nucleic acid drug which comprises adding the nucleic acid drug side effect reducing agent according to the present disclosure to the nucleic acid drug, and a present invention for reducing the side effect-inducing property of the nucleic acid drug.
  • the use of the disclosed oligopeptides is also provided.
  • the coexistence of the nucleic acid drug side effect reducing agent according to the present disclosure significantly reduces the side effects of the nucleic acid drug as a class effect.
  • Example 1 a double-stranded nucleic acid complex composed of an antisense oligonucleoside and a platelet-bound complementary strand is mixed with each of Dab8, Dab10, or Dab12 as a nucleic acid drug side effect reducing agent, and then the double-stranded nucleic acid is used.
  • a single dose of the nucleic acid complex was administered to mice to assess antisense effects and APTT (activated partial thromboplastin time), platelet count and brain pathology at various sites in the mouse brain and liver.
  • APTT activate partial thromboplastin time
  • the structures of Dab8, Dab10 and Dab12 are shown below.
  • these names are the names about the repeating structure after the acetyl tyrosine-glycine-glycine of the amino terminal part, together with the number of repetitions.
  • a double-stranded nucleic acid agent consisting of an antisense oligonucleoside targeting malat1 which is a long non-coding RNA and a complementary strand in which cholesterol is bound to the 5'end is used as a nucleic acid drug side effect reducing agent.
  • a single dose was administered to mice, and an experiment was conducted to evaluate the effect of inhibiting the expression of malat1 RNA in the brain in vivo.
  • the experiment was carried out with the amount ratio of the double-stranded nucleic acid complex: nucleic acid drug side effect reducing agent (Dab8, Dab10 or Dab12) to be 1: 1 or 1: 2 in molar ratio.
  • the activated partial thromboplastin time (aPTT), platelet count, cerebral pathology, and sedative effect immediately after administration were also evaluated. The specific contents of the experiment will be described below.
  • This LNA / DNA gapmer was a mouse malat1 non-coding RNA (GenBank accession). It had a complementary base sequence at positions 1316-1331 of No. NR_002847). Annealing of the first chain and the second chain was carried out by equimolar amounts after dissolving the first and second chains with PBS, respectively. The resulting solution was heated at 95 ° C. for 5 minutes, then cooled to 37 ° C. and held for 1 hour. The prepared double-stranded nucleic acid complex is referred to as Chol-HDO.
  • the side effect reducing agents (Dab8, Dab10 or Dab12) were all dissolved in PBS to a final concentration of 5 mM.
  • the underlined uppercase letter represents LNA, where " C " represents 5-methylcytosine LNA, the lowercase letter represents DNA, and the asterisk represents the phosphorothioate bond that connects the nucleosides.
  • uppercase letters represent RNA
  • underlined lowercase letters represent 2'-O-methylated RNA
  • asterisks represent phosphorothioate bonds that connect nucleosides
  • Chol represents cholesterol
  • a solution of the above double-stranded nucleic acid complex concentration of the double-stranded nucleic acid complex is 800 ⁇ M
  • a solution of a nucleic acid drug side effect reducing agent Dab8, Dab10 or Dab12 are combined with the double-stranded nucleic acid complex and the nucleic acid drug side effect.
  • mice injected with only the PBS solution and mice injected with only the above Chol-HDO solution were also prepared.
  • a group was also prepared in which PEG200 (manufactured by Sigma-Aldrich) was added so that the final concentration was 10% by mass in order to increase the solubility of the mixed solution of Chol-HDO and a nucleic acid drug side effect reducing agent (Dab8, Dab10 or Dab12). ..
  • the molar ratio represents a double-stranded nucleic acid complex: nucleic acid drug side effect reducing agent.
  • Mouse group 1 PBS solution
  • Mouse group 2 Chol-HDO solution
  • Mouse group 3 A mixture of Chol-HDO and Dab8 (molar ratio 1: 1)
  • Mouse group 4 Mixed solution of Chol-HDO and Dab8 (molar ratio is 1: 2)
  • Mouse group 5 Mixed solution of Chol-HDO and Dab8 (molar ratio: 1: 2) + 10% by mass PEG200
  • Mouse group 6 A mixture of Chol-HDO and Dab10 (molar ratio 1: 1)
  • Mouse group 7 Mixed solution of Chol-HDO and Dab10 (molar ratio is 1: 2)
  • Mouse group 8 Mixed solution of Chol-HDO and Dab10 (molar ratio: 1: 2) + 10% by mass PEG200
  • Mouse group 9 Mixed solution of Chol-HDO and Dab12 (molar ratio 1: 1)
  • Mouse group 10 Mixed solution of Chol-HDO and Dab12 (m
  • mice Seventy-two hours after the above intravenous injection, PBS was perfused into mice, after which the mice were dissected and the cerebral cortex, cerebellum, striatum, liver, spinal cord, and brain stem were collected separately. Subsequently, mRNA was extracted from each tissue according to a protocol using a high-throughput fully automatic nucleic acid extractor MagNA Pure 96 (manufactured by Roche Life Sciences). From the extracted mRNA, cDNA was synthesized by using the reverse transcription reaction reagent PrimeScript RT Master Mix (TAKARA-Bio) according to the protocol.
  • TAKARA-Bio reverse transcription reaction reagent PrimeScript RT Master Mix
  • RT-PCR quantitative RT-PCR was carried out by Light Cycler 480 Probes Master (manufactured by Roche Life Science), which is a reagent for RT-PCR, and Light Cycler 480 (manufactured by Roche Life Science), which is an RT-PCR device.
  • the primers used in quantitative RT-PCR were products designed and manufactured by Thermo Fisher Scientific for various gene numbers.
  • the amplification conditions were as follows: A cycle consisting of 95 ° C. for 15 seconds, 60 ° C. for 30 seconds, and 72 ° C. for 1 second was defined as one cycle, and the cycle was repeated 40 cycles.
  • the APTT value in each group was 21.1 seconds in the mouse group 1, whereas it was as long as 149 seconds in the mouse group 2. This represents the side effect of prolonging APTT by administration of nucleic acid drugs.
  • a remarkable improvement was observed in the mouse group 3 to which the nucleic acid drug side effect reducing agent according to the present disclosure was administered to 86.4 seconds, and in the mouse group 4 to 33.1 seconds.
  • mice (Observation of sedative effect) The behavior of the mice was observed from immediately after the intravenous injection to 1.5 hours after the above intravenous injection. The evaluation was performed according to the following criteria. A: There is no abnormality in the behavior of the mouse, and it is actively active. B: Mouse behavior is slightly reduced. C: Mouse behavior is greatly reduced. D: The mouse hardly moves. The results are shown in Table 4 below.
  • Example 2 (Further in vivo experiment: Evaluation of APTT improvement effect in mouse in vivo by addition of a double-stranded nucleic acid complex consisting of an antisense oligonucleotide and a cholesterol-bound complementary strand and a nucleic acid drug side effect reducing agent)
  • a double-stranded nucleic acid agent consisting of an antisense oligonucleotide targeting malat1 and a cholesterol-binding complementary strand described in the section of "Preparation of double-stranded nucleic acid complex" of Example 1 reduces the side effects of nucleic acid drugs according to the present disclosure.
  • An experiment was conducted in which the agents were mixed, administered to mice, and blood was collected after a lapse of a predetermined time to evaluate the improvement of the APTT prolonging effect of nucleic acids. The details of the experiment are as follows.
  • the mouse used in the experiment was a 6-7 week old male Crl: CD1 mouse weighing 20 g.
  • a solution of the above double-stranded nucleic acid complex concentration of the double-stranded nucleic acid complex is 800 ⁇ M
  • a solution of a nucleic acid drug side effect reducing agent Dab8, Dab10 or Dab12
  • Dab8, Dab10 or Dab12 a nucleic acid drug side effect reducing agent
  • the molar ratio represents a double-stranded nucleic acid complex: nucleic acid drug side effect reducing agent.
  • Mouse group 1 Chol-HDO solution
  • Mouse group 2 A mixed solution of Chol-HDO and Dab8 (molar ratio is 1: 1)
  • Mouse group 3 Mixed solution of Chol-HDO and Dab8 (molar ratio is 1: 2)
  • Mouse group 4 A mixture of Chol-HDO and Dab10 (molar ratio 1: 1)
  • Mouse group 5 Mixed solution of Chol-HDO and Dab10 (molar ratio is 1: 2)
  • Mouse group 6 A mixture of Chol-HDO and Dab12 (molar ratio 1: 1)
  • Mouse group 7 Mixed solution of Chol-HDO and Dab12 (molar ratio is 1: 2)
  • the ratio (T / S) of the base length T of the nucleic acid contained in the nucleic acid drug to the number S of the amino acid residues of the formula (I) in the specific oligopeptide region, and the molar of the nucleic acid drug side effect reducing agent is (T / S). It is shown in Table 5 below.
  • Example 2 The results of Example 2 are shown in the graphs of FIGS. 4 to 6.
  • Chol-HDO 20 mg / kg represents mouse group 1
  • "+ Dab8 1: 1” represents mouse group 2
  • "+ Dab8 1: 2” represents mouse group 3.
  • "+ Dab10 1: 1” represents mouse group 4
  • "+ Dab10 1: 2” represents mouse group 5
  • "+ Dab12 1: 1” represents mouse group 6
  • “+ Dab12 1: 2” represents mouse group 7. .
  • error bars indicate standard errors. In the results of Example 2, the following was observed.
  • the aPTT value (mean) 30 minutes after administration was 51.4 seconds
  • the aPTT value 120 minutes later was 42.6 seconds
  • the aPTT value 240 minutes after administration was , 29.3 seconds
  • the extension of aPTT was observed (FIGS. 4 to 6).
  • the aPTT value (mean) 30 minutes after administration in each group was 51.4 seconds in the mouse group 1, 42.2 seconds in the mouse group 4, 30.9 seconds in the mouse group 5, and mice. It was 43.1 seconds in group 6 and 26.7 seconds in mouse group 7 (Fig. 4).
  • the aPTT value (mean) 120 minutes after administration in each group was 42.6 seconds in the mouse group 1, 39.0 seconds in the mouse group 2, 33.6 seconds in the mouse group 3, and the mouse group. 4 was 40.4 seconds, mouse group 5 was 22.4 seconds, mouse group 6 was 35.6 seconds, and mouse group 7 was 18.7 seconds (FIG. 5).
  • the aPTT value (mean) 240 minutes after administration in each group was 29.3 seconds in the mouse group 1, 28.0 seconds in the mouse group 4, and 17.7 seconds in the mouse group 5. It took 26.4 seconds in the mouse group 6 and 17.5 seconds in the mouse group 7 (Fig. 6).
  • the dose-dependent shortening effect of aPTT on Dab8 / 10/12 was analyzed by Williams' test. As a result, the following was found. -Compared with the mouse group 1, in the mouse group 2 to which Dab8 was added 1: 1 and the mouse group 3 to which Dab8 was added 1: 2, the shortening of aPTT 120 minutes after administration was not so large ( 4 to 6). -Compared with the mouse group 1, in the mouse group 4 to which Dab10 was added 1: 1 and the mouse group 5 to which Dab10 was added 1: 2, 30 minutes after administration, 120 minutes after administration, and 240 minutes after administration.
  • Mouse group 1 vs mouse group 4: P 0.004, mouse group 1 vs mouse group 4: P ⁇ 0.001 240 minutes after administration (FIG. 6).
  • the aPTT shortening effect tended to be greater in a dose-dependent manner of Dab10 (that is, when the molar ratio of Chol-HDO: Dab10 was 1: 1 than when the molar ratio was 1: 2). (FIGS. 4 to 6).
  • aPTT was significantly shortened (mouse group 1 vs mouse group 6: P ⁇ 0.001 30 minutes after administration, mouse group 1 vs mouse group 7: P ⁇ 0.001 30 minutes after administration (Fig.). 4), mouse group 1 vs mouse group 6: P ⁇ 0.001 120 minutes after administration, mouse group 1 vs mouse group 7: P ⁇ 0.001 (Fig. 5) 120 minutes after administration, 240 minutes after administration.
  • the aPTT shortening effect is as large as the amount of Dab12 added. (That is, the case where the molar ratio of Chol-HDO: Dab12 was 1: 1 was higher than the case where the molar ratio was 1: 2).
  • the shortening effect of each aPTT of Dab8, Dab10, and Dab12 was analyzed by Williams' test.
  • Mouse group Dab8 was added 1: 1 to mouse group 2
  • Dab10 was added 1: 1 to mouse group 4
  • Dab12 was added 1: 1 to mouse group 6.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • General Health & Medical Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Engineering & Computer Science (AREA)
  • Organic Chemistry (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Animal Behavior & Ethology (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Genetics & Genomics (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Epidemiology (AREA)
  • Molecular Biology (AREA)
  • Biomedical Technology (AREA)
  • Biophysics (AREA)
  • Wood Science & Technology (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • General Chemical & Material Sciences (AREA)
  • Immunology (AREA)
  • Biotechnology (AREA)
  • General Engineering & Computer Science (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Biochemistry (AREA)
  • Zoology (AREA)
  • Microbiology (AREA)
  • Toxicology (AREA)
  • Inorganic Chemistry (AREA)
  • Plant Pathology (AREA)
  • Physics & Mathematics (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
PCT/JP2020/025093 2019-06-28 2020-06-25 核酸医薬副作用軽減剤、該核酸医薬副作用軽減剤を含む医薬組成物、並びに核酸医薬の副作用惹起性を軽減する方法 Ceased WO2020262555A1 (ja)

Priority Applications (1)

Application Number Priority Date Filing Date Title
JP2021527744A JPWO2020262555A1 (https=) 2019-06-28 2020-06-25

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
JP2019120908 2019-06-28
JP2019-120908 2019-06-28

Publications (1)

Publication Number Publication Date
WO2020262555A1 true WO2020262555A1 (ja) 2020-12-30

Family

ID=74061653

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/JP2020/025093 Ceased WO2020262555A1 (ja) 2019-06-28 2020-06-25 核酸医薬副作用軽減剤、該核酸医薬副作用軽減剤を含む医薬組成物、並びに核酸医薬の副作用惹起性を軽減する方法

Country Status (2)

Country Link
JP (1) JPWO2020262555A1 (https=)
WO (1) WO2020262555A1 (https=)

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2014148620A1 (ja) * 2013-03-21 2014-09-25 国立大学法人東京医科歯科大学 二重鎖核酸結合剤、当該結合剤－二重鎖核酸複合体、当該複合体を含有する医薬品組成物、及び、当該複合体の製造方法
WO2019013255A1 (ja) * 2017-07-11 2019-01-17 学校法人東京薬科大学 核酸送達用組成物及び核酸含有組成物

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2014148620A1 (ja) * 2013-03-21 2014-09-25 国立大学法人東京医科歯科大学 二重鎖核酸結合剤、当該結合剤－二重鎖核酸複合体、当該複合体を含有する医薬品組成物、及び、当該複合体の製造方法
WO2019013255A1 (ja) * 2017-07-11 2019-01-17 学校法人東京薬科大学 核酸送達用組成物及び核酸含有組成物

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
HARA RINTARO IWATA, MAEDA YUSUKE, FUJIMAKI HARUNA, WADA TAKESHI: "Enhancement in RNase H activity of a DNA/RNA hybrid duplex using artificial cationic oligopeptides", CHEM COMMUN, vol. 54, no. 61, 2018, pages 8526 - 8529, XP055778726 *
MAEDA YUSUKE, IWATA HARA RINTARO, NISHINA KAZUTAKA, YOSHIDA-TANAKA KIE, SAKAMOTO TAIICHI, YOKOTA TAKANORI, WADA TAKESHI: "Artificial cationic peptides that increase nuclease resistance of siRNA without disturbing RNAi activity", NUCLEOSIDES , NUCLEOTIDES AND NUCLEIC ACIDS, vol. 38, no. 5, 2019, pages 338 - 348, XP055778723 *
MAEDA, Y ET AL.: "S ynthesis and properties of cationic oligopeptides with different side chain lengths that bind to RNA duplexes", BIOORGANIC & MEDICINAL CHEMISTRY, vol. 21, no. 7, 2013, pages 1717 - 1723, XP029002608, DOI: 10.1016/j.bmc.2013.01.053 *

Also Published As

Publication number Publication date
JPWO2020262555A1 (https=) 2020-12-30

Similar Documents

Publication Publication Date Title
JP7423015B2 (ja) キメラ２重鎖核酸
CN111107853B (zh) 用于抑制载脂蛋白C-III (APOC3)的表达的RNAi试剂和组合物
CN113164509B (zh) 用于抑制17β-HSD 13型(HSD17B13)表达的RNAi试剂、其组合物和使用方法
WO2020196662A1 (ja) 二本鎖核酸複合体及びその使用
JP6440170B2 (ja) キメラ一本鎖アンチセンスポリヌクレオチドおよび二本鎖アンチセンス剤
CN115397436B (zh) 用于抑制PNPLA3表达的RNAi剂、其药物组合物和使用方法
WO2018062510A1 (ja) オーバーハングを有する二本鎖核酸複合体
WO2021070959A1 (ja) 修飾ヘテロ核酸
JP6934695B2 (ja) 核酸医薬とその使用
WO2020262555A1 (ja) 核酸医薬副作用軽減剤、該核酸医薬副作用軽減剤を含む医薬組成物、並びに核酸医薬の副作用惹起性を軽減する方法
WO2024192379A1 (en) Rnai agents for inhibiting expression of mitochondrial amidoxime reducing component 1 (marc1), pharmaceutical compositions thereof, and methods of use
HK40084771B (zh) 用於抑制PNPLA3表达的RNAi剂、其药物组合物和使用方法
KR20250164214A (ko) 아탁신-2(ATXN2)의 발현을 억제하기 위한 RNAi 제제, 이의 조성물, 및 사용 방법
HK40084771A (en) Rnai agents for inhibiting expression of pnpla3, pharmaceutical compositions thereof, and methods of use
EA047975B1 (ru) РНКи АГЕНТЫ ДЛЯ ИНГИБИРОВАНИЯ ЭКСПРЕССИИ 17β-HSD ТИПА 13 (HSD17B13), ИХ КОМПОЗИЦИИ И СПОСОБЫ ПРИМЕНЕНИЯ

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 20831482

Country of ref document: EP

Kind code of ref document: A1

DPE1 Request for preliminary examination filed after expiration of 19th month from priority date (pct application filed from 20040101)
ENP Entry into the national phase

Ref document number: 2021527744

Country of ref document: JP

Kind code of ref document: A

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 20831482

Country of ref document: EP

Kind code of ref document: A1