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WO1996003133A1 - Nouveau medicament anti vih - Google Patents

Nouveau medicament anti vih Download PDF

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Publication number
WO1996003133A1
WO1996003133A1 PCT/JP1995/001456 JP9501456W WO9603133A1 WO 1996003133 A1 WO1996003133 A1 WO 1996003133A1 JP 9501456 W JP9501456 W JP 9501456W WO 9603133 A1 WO9603133 A1 WO 9603133A1
Authority
WO
WIPO (PCT)
Prior art keywords
hydroxyl group
formula
active ingredient
group
represented
Prior art date
Application number
PCT/JP1995/001456
Other languages
English (en)
Japanese (ja)
Inventor
Akira Kaji
Hideko Kaji
Original Assignee
Akira Kaji
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
Priority claimed from JP20003194A external-priority patent/JPH0834737A/ja
Priority claimed from JP20003294A external-priority patent/JPH0834738A/ja
Priority claimed from JP20002994A external-priority patent/JPH0834735A/ja
Priority claimed from JP20003094A external-priority patent/JPH0834736A/ja
Priority claimed from JP23831494A external-priority patent/JPH0859478A/ja
Priority claimed from JP23831794A external-priority patent/JPH0859481A/ja
Priority claimed from JP23831694A external-priority patent/JPH0859480A/ja
Priority claimed from JP23831594A external-priority patent/JPH0859479A/ja
Application filed by Akira Kaji filed Critical Akira Kaji
Priority to EP95926009A priority Critical patent/EP0724882A4/fr
Publication of WO1996003133A1 publication Critical patent/WO1996003133A1/fr

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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
    • A61K31/7105—Natural ribonucleic acids, i.e. containing only riboses attached to adenine, guanine, cytosine or uracil and having 3'-5' phosphodiester links
    • 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
    • A61K31/7125—Nucleic acids or oligonucleotides having modified internucleoside linkage, i.e. other than 3'-5' phosphodiesters
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
    • C07H21/00—Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids

Definitions

  • the present invention relates to a phosphodiester-binding type or a hoshortiester-binding type that exhibits excellent antiviral activity against diseases caused by HIV (Human Immunodeficiency Virus).
  • HIV Human Immunodeficiency Virus
  • the present invention relates to an anti-HIV drug containing a ribonucleotide as an active ingredient.
  • the present inventor has conducted various studies on the reduction of the molecular weight of an oligonucleotide compound having an inhibitory effect on the growth of HIV, and has first made a short 4 to 8 nucleotides. It was found that chain oligodoxyguanylic acid has an HIV growth inhibitory effect.
  • the present invention also provides a phosphodiester-bonded oligoribonucleotide represented by the formulas (I) to (XX ⁇ I) described in the following items (a) to (e). Or one or more selected from hoshortiester-bonded oligoribonucleotides as an active ingredient. It provides anti-HIV drugs.
  • E E represented by any of the following formulas (I) to (IV) An anti-HIV agent characterized in that it contains one or more selected from among the phosphodiester-binding oligoribonucleotides as an active ingredient. .
  • G represents a guanosine structure
  • ⁇ represents a phosphoric acid residue forming an ester bond with an adjacent G
  • ⁇ represents the 3 ′
  • G Forms a monoester bond with the hydroxyl group at the 3'-position of the terminal G
  • each ⁇ located between the two Gs is the G on the left side in the formula. Is an ester bond with the 3'-hydroxyl group, and G on the right-hand side is an ester bond with the 5'-hydroxyl group.
  • each G and p are as defined above, and the left terminal p is the 5'-position of the terminal G. It forms a monoester bond with a hydroxyl group.
  • G s G s G s G p (XI) G s G s G s G s G p (XI I) G s G s G s G s G s G p (XI II) G s G s G s G s G s G sp (IV) G s G s G s G s G s G s G s G p (XV) (wherein each G and p represent the same as above, and s is The s represents a phosphorothiophosphoric acid residue that forms a phosphorothioester bond with an adjacent G, and each s is 3 'to the G on the left side in the formula. The hydroxyl group and the right-hand side G form an ester bond with the 5′-position hydroxyl group, respectively).
  • (Ii) one selected from the phosphoholoester-bonded oligoribonucleotides represented by any of the following formulas (XVI) to (IXX) Or, an anti-HIV agent characterized by containing two or more kinds as an active ingredient.
  • oligoribonucleotides represented by the above formulas (I) to (XXIII), which are the active ingredients of the anti-HIV agent of the present invention, are generally used for the synthesis of oligoribonucleotides.
  • each internucleotide is subjected to ammonia treatment by a triester. It converts to physical strength, and is extracted from the support. At the same time, the base is deprotected, and the tetra-N-butylammonium derivative is removed.
  • a method in which the hydroxyl group at the 2'-position is deprotected by a light treatment to obtain the desired 3'-terminal phosphorylated oligoribonucleotide.
  • guanosine with a protected functional group is applied to a support such as control-bore glass (CPG).
  • CPG control-bore glass
  • 4'-dimethoxytrityl group which is a protecting group for the 5'-hydroxyl group of this nucleotide, to trichloroacetic acid, etc.
  • Protected amidite reagent (5'-o—dimethoxycitryl) 1-2'-o— (t-butyltin dimethyltrisilyl) ribonucleic acid Leoside 3'-N, N — Condensates diisobrovinore (cyanethyl) phosphoramigite. Thereafter, the phosphorus portion is oxidized using iodine to lead to a phosphoric acid triester body. Next, the 5'-protecting group (DMTR) of this guanylic acid is removed with trichloroacetic acid, and the second amidate protecting the functional group of guanylic acid is removed. The reagents are condensed as above.
  • Amidite reagent in which the amino group of the acid base is protected with an isobutyl group, (5'-o-dimethoxycitryl 1 2 ')-o-1 ( t — Butinore di-methyl thiol) Ribonucleoside 3'-N, N — Diisopropynole (cyanoethyl) Condensed using tetrazole as a condensing agent.
  • the unreacted 5'-hydroxyl group is acetylated with acetic anhydride, N-methylimidazole, or the like.
  • the rind is sulfurized using tetraethyl uranium disulphide, which leads to the hoshorochiotriester body.
  • a phosphorylation reagent (2—cyanoethoxy) is used.
  • the phosphine is condensed (see Tetrahedron Letters, supra), and the terminal phosphorus is oxidized to form a phosphoric acid triester using ammonium chloride.
  • the processing converts the triester into a diester, which is cleaved from the support and deprotected at the same time as the base.
  • the hydroxyl group at the 2'-position is deprotected by the treatment with dimethylphenol, and the phosphorylated phosphorylated phosphate at the 5'-terminal is targeted.
  • a force method that makes it a polygon.
  • the trimethyl group is removed with trichloroacetic acid or the like, and the amidate reagent in which the amino group of the guanylate group is protected with the isopropyl group, (5 '— o — Dimethyl citrine) 1 2'-o-(t-butyl dimethylsilyl) Ribonucleoside 3 '-N, N- Condensation of disopropinole (cyanoethyl) phosphamide using tetrazole as a condensing agent. To prevent this, the unreacted 5'-hydroxyl group is acetylated with acetic anhydride, N-methylimidazole, etc.
  • tetraethylate Sulfuration of the phosphorous part is performed using a urethium sulfide and the sulfide is led to a phosphoric acid triester body.
  • ammonia processing is used to remove each nucleotide from the triester body to the polyester.
  • the product is converted to a thiol form, cut out from the support and deprotected at the same time as the base, and treated at the 2'-position by tetra-N-butylammonium fluoride treatment.
  • a method for deprotecting hydroxyl groups to obtain the target oligoribonucleotide is performed using a urethium sulfide and the sulfide is led to a phosphoric acid triester body.
  • 96--4 cells were spread on a 96-well microplate with a volume of 6 x 10 4 cells / well, and then used for AIDS virus (HIV-1 (I ⁇ B; about 50 TCI). (50% cultured cell infection concentration) / well] and cultured for 5 days at 37 ° C and 5% carbon dioxide. To determine the inhibition of cell damage due to infection, the cells were cultured for 5 days. It was determined by measuring the cells by the MTT method.
  • G p G p G p G 100 // g / ml or more
  • phos which is an active ingredient of the anti-HIV agent of the present invention
  • Anti-HIV activity is observed even with 2 nucleotides of phospho- or phospho-ester-bound oligoribonucleotides, while excellent anti-HIV activity is observed with 3 to 8 nucleotides. It was shown to have HIV effects. Furthermore, none of the oligoribonucleotides showed cytotoxicity at a concentration of 100 g / ml.
  • phosphodiester-linked oligoguanic acid in which the terminal G was not phosphorylated had no effect at all with a few bases, and showed a slight effect with four bases. In addition, no effect was observed with any of the four bases of 3′-terminal phosphorylated phosphodiester-bonded oligoadenylic acid, citidic acid, and peridinic acid.
  • the 3'-terminal or 5'-terminal phosphorylated phosphodiester-bonded type and the phosphoroester-type bonded oligoguanylic acid it is effective. The gender is recognized to be outstanding.
  • the novel anti-HIV agent according to the present invention is particularly useful as a therapeutic agent for retroviral infections such as AIDS, and is formulated into a tablet using a pharmaceutical carrier, and is used as a tablet. It can be made into oral dosage forms such as capsules, capsules, granules, syrups, and powders, and parenteral dosage forms such as injections, infusions, and suppositories.
  • the dosage form is not specified, and an appropriate dosage form may be selected and used as needed, such as tablets, capsules, and granules. And parenteral administration agents such as injections, suppositories and the like.
  • the amount required as an oral preparation varies depending on the age, weight, and degree of disease of the patient, and the dosage for adults is usually 0.1 g to 6 g divided into several times a day. Take it.
  • oral preparations such as tablets, capsules, granules, etc.
  • starch lactose, sucrose, mannitol, carboxymethyl Cellulose and inorganic salts can be used, and are produced according to a conventional method.
  • a binder In this type of preparation, a binder, a disintegrant, a surfactant, a lubricant, a fluidity enhancer, a flavoring agent, a coloring agent, a fragrance, etc. are appropriately selected and used in addition to the above-mentioned excipients. You can do it. Specific examples are as follows.
  • Starch dextrin, arabia rubber powder, gelatin, hydroxypropinorestin, methinoresenolose, carboxy Methylcellulose sodium, hydroxypropylcellulose, crystal cellulose, ethylcellulose, Livininore Senororose, McGorgo Norre.
  • Talc roux, hydrogenated vegetable oil, sucrose fatty acid ester, magnesium stearate, polyethylene glycol.
  • the oligoribonucleotides represented by the formulas (I) to (XXm) according to the present invention include suspensions, emulsion liquids, syrups, and elixirs. It can also be administered as a preparation, and each of these dosage forms can contain a flavoring agent and a coloring agent.
  • the amount required as a parenteral preparation varies depending on the age, weight, and degree of the disease of the patient, but it is usually 1 to 1 for the above-mentioned oligoribonucleotides per day for adults. It is about 00 mg and is administered by intravenous injection, intravenous drip infusion, subcutaneous injection, or intramuscular injection.
  • This parenteral preparation can be manufactured by a conventional method.
  • the diluents are generally distilled water for injection, physiological saline, aqueous solution of glucose, vegetable oil for injection, sesame oil, racquet oil, soybean oil, and grease. Corn oil, propylene glycol, polyethylene glycol, etc. can be used.
  • fungicides preservatives, and stabilizers can be added as needed.
  • this parenteral preparation is filled into a vial or the like, then frozen, the water is removed by ordinary freeze-drying technology, and the product is lyophilized immediately before use.
  • the solution can be reconstituted from the lyophilized product at the time of use.
  • an isotonic agent, a stabilizer, a preservative, an analgesic agent and the like can be appropriately added.
  • parenteral preparations examples include liquid preparations for external use, coating preparations such as soft blue, suppositories for rectal administration, etc., all of which can be manufactured by a conventional method. it can.
  • Example 1 Example 1
  • This one tablet contains 20 mg of GpGpGpGp. This tablet is taken in several doses of 4 to 8 tablets daily for an adult.
  • magnesium stearate 0.5% by weight magnesium stearate
  • This one tablet contains 20 mg of pGpGpGpG. Adults take this tablet in 4 divided doses 8 times a day for adults. Used.
  • novel anti-HIV agent according to the present invention is effective for the treatment of diseases caused by HIV (AIDS virus), AIDS and ARC.

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  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Biochemistry (AREA)
  • Molecular Biology (AREA)
  • General Health & Medical Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • Epidemiology (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Medicinal Chemistry (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Organic Chemistry (AREA)
  • Engineering & Computer Science (AREA)
  • Biotechnology (AREA)
  • Genetics & Genomics (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Abstract

Médicament anti VIH contenant un principe actif comprenant au moins un constituant sélectionné à partir d'oligoribonucléotides à liaisons phosphodiester et phosphorothioester de di- à octabase, représentés par les homologues à tétrabase représentés par les formules suivantes (III), (VII), (XI), (XVII) et (XXI). G p G p G p G p (III), p G p G p G p G (VII), G s G s G s G p (XI), p G s G s G s G (XVII), G s G s G s G (XXI), dans lesquelles G représente une structure guanosine, p représente un résidu de phosphate constituant une liaison ester comportant un G contigu et s représente un résidu de phosphorothiophosphate constituant une liaison phosphorothioester comportant un G contigu.
PCT/JP1995/001456 1994-07-22 1995-07-21 Nouveau medicament anti vih WO1996003133A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
EP95926009A EP0724882A4 (fr) 1994-07-22 1995-07-21 Nouveau medicament anti vih

Applications Claiming Priority (17)

Application Number Priority Date Filing Date Title
JP6/200029 1994-07-22
JP6/200031 1994-07-22
JP6/200030 1994-07-22
JP20003194A JPH0834737A (ja) 1994-07-22 1994-07-22 新規抗ｈｉｖ剤
JP20003294A JPH0834738A (ja) 1994-07-22 1994-07-22 新規抗ｈｉｖ剤
JP20002994A JPH0834735A (ja) 1994-07-22 1994-07-22 新規抗ｈｉｖ剤
JP6/200032 1994-07-22
JP20003094A JPH0834736A (ja) 1994-07-22 1994-07-22 新規抗ｈｉｖ剤
JP6/238313 1994-08-26
JP23831494A JPH0859478A (ja) 1994-08-26 1994-08-26 新規抗ｈｉｖ剤
JP23831794A JPH0859481A (ja) 1994-08-26 1994-08-26 新規抗ｈｉｖ剤
JP6/238315 1994-08-26
JP6/238316 1994-08-26
JP23831694A JPH0859480A (ja) 1994-08-26 1994-08-26 新規抗ｈｉｖ剤
JP6/238317 1994-08-26
JP6/238314 1994-08-26
JP23831594A JPH0859479A (ja) 1994-08-26 1994-08-26 新規抗ｈｉｖ剤

Publications (1)

Publication Number Publication Date
WO1996003133A1 true WO1996003133A1 (fr) 1996-02-08

Family

ID=27573416

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/JP1995/001456 WO1996003133A1 (fr) 1994-07-22 1995-07-21 Nouveau medicament anti vih

Country Status (1)

Country Link
WO (1) WO1996003133A1 (fr)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0743318A4 (fr) * 1994-12-02 1998-09-02 Akira Kaji Nouvel agent anti-vih

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
KIM, S. et al., "Antiviral Effect of Phosphorothioate Oligodeoxyribonucleotides Complementary to Human Immunodefficiency Virus", BIOORG. MED. CHEM., (1995), 3(1), p. 49-54. *
See also references of EP0724882A4 *
SHIBAHARA, S. et al., "Inhibition of Human Immunodeficiency Virus (HIV-1), Replication by Synthetic Oligo-RNA Derivatives", NUCLEIC ACIDS RES., (1989), 17(1), p. 239-252. *

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0743318A4 (fr) * 1994-12-02 1998-09-02 Akira Kaji Nouvel agent anti-vih

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