SA92120445B1 - Use of deoxy-3 "thiacytidine 5-fluoro- 2" for the treatment of hepatitis B virus - Google Patents

Abstract

THE USE OF A 1,3-OXATHIOLANE NUCLEOSIDE ANALOGUE AND PHARMACEUTICALLY ACCEPTABLE DERIVATIVES THEREOF FOR THE TREATMENT OF HEPATITIS B VIRUS INFECTIONS IS DISCLOSED. PHARMACEUTICAL FORMULATIONS ARE ALSO PROVIDED.

Description

USE OF 5-FLUORO-2'-DEOXY-3'-THIACYTIDINE FOR "Treatment of Hepatitis B Virus" FULL DESCRIPTION BACKGROUND OF THE INVENT The present invention relates to the use of a derivative 1,7-hydroxymethyl-0,7- mo OY SSS yl-cytosine 1-2-hydroxymethyl-1,3-oxathiolan-5-yl-cytosine and their physiologically functional derivatives for the treatment of viral infections of Hepatitis B Hepatitis B causes hepatitis C virus HBV is a viral germ Of great importance in the world wide Hepatitis C disease is common in Asian countries and predominates in Africa in sub-Saharan Africa The virus is associated with primary carcinoid tumors of cal cells and is believed to cause 7850 liver cancers in: the world In the United States it is treated in hospitals more Out of ten thousand patients suffer from hepatitis C 1131 each year; Lay a YOu dies on average of the deadly disease—and in the United States the disease is carried by an estimated half million to one million people. carriers of the disease virus, and it often develops into cirrhosis of the liver, and it is estimated that 900,810 people suffer from SUH-related cirrhosis of the liver each year in the United States; 1,000 people are likely to die: from hepatitis C virus-related liver cancer. Even with the existence of a vaccine against hepatitis virus SUS 1 worldwide; The need for effective anti-HCV compounds Cand SY is urgent and will continue to be. It will not work for a large number of HIV carriers who are infected with it, estimated at 770 million people in the world, to receive a protective vaccine, because they will be constantly exposed to the seriousness of liver disease induced by the hepatitis virus. Carriers of this disease help as a source of infection for individuals exposed to it, especially in endemic areas, or other groups exposed to its risks © _such as groups that abuse drugs or homosexuals. Thus, there is an urgent need for effective antiviral agents; To eliminate acute infection and reduce disease progression to hepatocellular carcinoma. Clinical effects of HCV range from headache to fever to a general “0” feeling of illness to nausea; to vomiting; to lack of appetite for food; and abdominal pain. ASE virus is eliminated by the immune response following a period of convalescent therapy lasting several weeks to months in humans; However, the infection may be more severe, leading to acute and chronic liver disease, as mentioned above. And in the book: A second edition, Ed. Evans, A.S. 1982 Plenum Publishing Corporation, New York, Hepatitis C Virus Infection. 11 Chapter describes a set of 10; “- Oxathiolane TAY 877 European Patent Application Disclosure 0 of a Possible HIV Treatment for HIV Viruses.” ; Especially 1,3-0xathiolane nucleosides SY dy -1-nisotis niclosides Saying that the compound 7- hydroxymethyl-#-*- fluoro- generally die las 2-hydroxymethyl-5-5-fluorocytosin- 1-yl-1,3-oxathiolane oxaio lan But not declared. about him in particular”; in this specification; It is disclosed by the patent application ald $B and not the hepatitis C virus (HIV) in general for the treatment of 0

Sn i OY Aminopurine-4-beta-- IB TeX for European Ye 451 for the treatment of 2,6-diaminopurine-9-beta-D-2',3'-dideoxyribofuranoside ribovir atoside No. 94 (Gastroenterology Viral Infection; especially 11 [7]. In the scientific journal 2',3'-dideoxycytidine VEY HH 552 H. reveals the effect of CY co part using WO92 /14743 on hepatitis B virus 3 [Petsa.] ve: enantiomers and enantiomers 2-hydroxymethyl-55-fluorocytosin-1-yl-1,3-oxathiolane

HBV; HIV of it in the treatment of oxathiolane-*-yl-YO = f Escordia-7-1-018 a We surprisingly found that the ©1: derivative represented by the formula cis-1-2-hydroxymethyl-1 Sustained Cytosin-Eng 5-valve 3-0, - Z like ©: |

HO N | 0) 0: mother 7 pm

¢ Specifically 8[”-1,7-hydroxy-Fo = fe oxathiolan-*-yl-*-fluorocytosine cis-1-2-hydroxymethyl-1,3-oxathiolan-5-yl-5-fluorocytosine and its derivatives Physiologically, it has strong activity against hepatitis C virus 11317. Z Z It should be noted that the compound represented by formula 1 contains two chiral centers, and therefore it is 0 in the form of a pair of optical isomers, meaning “optically identical” 38 and their mixtures including racemic mixtures thus; The compound represented by Formula 0; may be composed of a cis cls or a trans isomer or mixtures of both each of the trans 5 cis isomers present in a single optical lil die form or their mixtures include conjugate mixtures . 1 All of these analogues and their mixtures include isotropic mixtures and 'tautomeric' forms of formula 1 are within the scope of the invention. Preferably similar to the compound Jed of formula 1. Chua gl detailed z and according to one of the features of the present invention; We are working on finding a compound with formula 1 or a salt amide eo or a pharmaceutically acceptable ester or a salt of the ester thereof as a drug for the treatment or prevention of hepatitis C infection. The drug can be used to treat or prevent hepatitis C virus infection in a host, for example; It belongs to mammals like humans; The method includes: Treating the host with an effective amount, Ladle, of the compound loaded with the formula, ester sf famide lo Jf 1, pharmacologically acceptable, or an ester salt thereof: a. The term “physiologically functional derivative” means a salt, amide, ester, or Pharmtically acceptable ester salt: of a compound of formula 1.z © The preferred esters according to the invention include carboxylic acid esters in which a non-carbonylic moiety is selected for the carbocylic acid part of the ester groups. straight or branched-chain alkyl; e.g. vo propyl a-propyl butyl two-thirds t-butyl n-butyl alkyl lie alkoxyalkyl amethoxymethyl arylalkyl For example; Ole aryloxyalkyl benzyl phenoxymethyl and aryl methyl phenoxy

0 . | : cphenyl sulfonate esters, Sia sulfonate esters, emethanesulfonyl sulfonate, esters of amino acids, Otis amino acid esters, L-valyl- or L-isoleucyl

for | OG dicarboxylic acid esters are semi-succinates. hemisuccinate and Al deal —'o or ali wh esters ©. S-mono-di or tri-phosphate esters may also be esterified by; CMa is an alcohol containing 1-71 carbon atoms or its reactant derivative; or by FY di Cg oy acyl glycerol Baume 2,3-010. Any day a saalkyl moiety of these esters contains carbon 33 18-1 and especially 1 to ; carbon atoms and includes the distinction which aryl moiety in these phenyl de sana esters substituted old) eg (JU by an alkyl halogen. contains)-£ a carbon atom or an alkoxy It contains (£7) a carbon or nitro atom The pharmaceutically acceptable amides mentioned above for the compound of formula 1 include derivatives in which the amino group cytosine amino 03105106 is present in the form -HNCOR Mis amide where R is an alkyl or aryl group containing 1-1 atoms phenyl «Sis carbon optionally substituted vo by an alkyl chalogen containing 1-; alkoxy carbon atoms containing 1- Nitro carbon or hydroxyl atoms: Examples of pharmaceutically acceptable salts according to the invention include basic salts, for example, derived from an appropriate base, such as the salts of the alkali metal Sle sodium alkali earth metal Magnesium Sa 122200865110170 and ammonium R 11.6 where X is an alkyl, EY x contains carbon atoms, and acceptable acid addition salts Wa a include organic carboxylic acids, such as acetic, dactic, tartaric, malic, isthionic, lactobionic, succinic, and organic sulfonic acids: cmethanesulfonic, ethanesulfonic cethanesulfonic benzene sulfonic cbenzenesulfonic and para-tubulin sulfonic p-toluenesulfonic vo | and inorganic acids (Jia) chydrochloric, sulfuric, phosphoric, and sulfamic acids.

oo . and the amount of compound cis represented by formula 1 also referred to herein in this full description of the invention as the "active ingredient" or salt; Pharmacologically acceptable amide or ester or a salt of the ester thereof that is required in medication to reach the desired effect and this amount depends on a number of factors; especially for a particular use; the nature of the compound used; method of administration and the condition of the patient. general; The appropriate dose ranges from ? to 171 mg per kilogram of body weight per user per day; Preferably in the range + to 0 mg per kg body weight per day; and in preference in the range of 0 to 10 mg per kg body weight per day. It is preferable to give the dose in parts twice; or three; or four; or five, six or more times at appropriate intervals throughout the day. The divided dose may be administered in unit doses; Contains; Ola at 156-01 eae > 0 preferably 000011-71 mg; 501-700 mg of active ingredient per unit dose form is preferred. in an ideal way; The active ingredient must be capped to achieve valuable plasma concentrations of the active ingredient ranging from about 1 VO to about 1 micromole micromol; Preferably about? to 50 μmol; With an preference of about “ to about 1 micromol. dda eg JB ve may be achieved by intravenous injection of a solution containing 1.1 to 75% of the active ingredient; optionally in brine; Or it is taken orally in the form of boluses, large tablets, each containing about 1 to about 100 mg / kg of the active ingredient. It may maintain desirable levels of blood by continuous infusions to create infusion rates of about 1.00 to about © mg/kg/hr or by intermittent infusion rates containing about 1.4 to ~V mg/kg 0 of the active ingredient. (a) in the process of manufacturing a drug according to the invention; referred to in this full description of the invention below by the term 'combination'; ideally and implicitly mixes the cis compound represented by formula 1 or its salt; a pharmacologically acceptable ester 'amide or an ester salt thereof herein in this full description of the invention as Sa active'" with 0 carriers or one or more acceptable excipients Wy a and Y° - other therapeutic agents. The combinations include pharmaceutical forms suitable for oral administration; anus: nose; Objectively, including pharmaceutical images placed through the skin and in the oral cavity and under the tongue or Valll

No or for intraesophageal administration containing solutions for subcutaneous and intramuscular injection; Intravenous and intradermal. The combinations may conveniently exist in unit doses; It may be prepared by any methods known in the field of pharmacy. These methods include the step of combining the active ingredient with a carrier comprising one or more of the components All and ©. Blends are generally prepared by homogeneously combining the active ingredient with liquid carriers, micro-fractionated solid carriers, or both and then; as necessary; product formation. Blends of the present invention suitable for oral administration may exist as separate units Jie capsules; single-dose sachets or tablets each containing: a predetermined amount of active ingredient and as a powder or granule; as a solution or suspension in an aqueous or anhydrous liquid; Or as a liquid oil-in-water emulsion or a liquid oil-in-water emulsion. The component may exist. It is also in the form of boluses, spits, or paste. Tablets may be made by pressing or forming; and optionally with one or more secondary components. Compressed tablets may be prepared by compressing the active ingredient in a suitable machine in a free-flowing form such as powder or granules; optionally mixing it with a binder, for example; povidone vo gelatin <hydroxypropylmethyl cellulose lubricant; inert diluting agent preservative A catalyst for disintegration such as povidone “sodium starch glycollate” cross-linked; Sodium carboxymethyl cellulose cross-linking or surfactant or dispersing agent. Formed tablets may be made by molding into molds in a suitable machine a mixture of powdered compound after wetting it with an inert liquid diluting agent. The pal AVL (optionally shat. or notched) may be tuned to allow a slow or controlled release of the active ingredient using Ole hydroxypropylmethyl cellulose in varying proportions to create the desired side of release. Tablets may optionally be provided with a dissolving coating. By enteric juice, to allow release of the active ingredient into the vo-gut and not the stomach.

A

Suitable combinations for oral use as described above may include de gana organic buffering agents designed to neutralize gastric acidity. You may choose these organizing materials from

Chl or inorganic substances such as mixed weak acids or bases; With their covalent salts, formulations suitable for surface oral administration include dressings containing the active ingredient in acacia gum or a base material containing an aromatic and flavoring agent; often sucrose and acacia gelatin and pastilla containing the active ingredient in an inert base material such as tragacanth or sucrose and acacia; mouthwashes; Containing the active ingredient glycerin and glycerin in a suitable liquid carrier. There may be combinations used for rectal administration as a suppository with a suitable base material, salicylate, including for example cocoa butter or salicylate. There may be combinations suitable for vaginal administration. such as vaginal suppositories and tampons; creams; gel gels; pastes; foams; or solutions for spraying by atomization containing in addition to

Co the active ingredient on suitable carriers as known in Appropriate combinations for intraesophageal administration include sterile, buffered, anhydrous injection solutions that may contain antioxidant agents; organizing factors; Bacteria-regulating agents, sterile aqueous candle suspensions, and dissolved salts that make the mixture consistent with the intended user's blood salts, and anhydrous agents that may include suspending agents and thickening agents; as systems that split fats or otherwise, that aim to combine compounds with components of the blood or one or more organs of the body; The mixtures may be contained in hermetically sealed containers containing a unit dose or multiple doses; example; ampoules, glass vials containing injection solution and small vials; It may be stored in a lyophilized condition so that © for injection; Before use in SLA only requires the addition of a sterile liquid carrier; For example; case. Solutions and suspensions for intravenous injection may be prepared from sterile powders, granules, and tablets of the type previously described. Among the preferred unit dose combinations, combinations containing a daily dose, a unit dose, or a split dose for administration throughout the day may be mentioned; As stated here above in this vo full description of the invention; or their appropriate parts of an active component.‎ m q . It must be understood that in addition to the above-mentioned components in particular; The blends according to the present invention may include other agents traditional in the field for the type of blend in question; and for example; Combinations suitable for oral administration may contain sweetening agents; and thickening agents, flavoring agents, and odorants. (Jia, others) The compound may be prepared according to the formula © for example by: > 0 Has the formula ?a 1,3-0xathiolane with 5-F-Cytosine A reactance of an optionally protected compound of 80 1 0 ; kK Da 1 (i)

Ls hydroxy or a hydrogen protecting group in which [4] is a hydrogen dus being a leaving group; or b is the reactance of a compound of formula b 1011141 a 1

HN 8 0 (AB) R,0 is 0 with an amino fluorinating agent, in which 18 is as defined above and 183 is a protecting group or “cytosine in the -8 position of the ring” fluorine helps insert an atom

Ca¥ c is the reactance of a compound of formula z ©

H.N.F

A si. 0 N:

R,0 0 1 yen ; 5 Vo

I.E

Where in it, 18 be as defined above with an agent that helps convert the oxo group 070 in: and any remaining protecting groups on the camino are removed to the uracil group put -; to the uracil ring, for example, by hydrolysis in the presence of an acid or a base to obtain the desired product. Examples of acyl-acetyls on hydroxy groups include a protecting group of for method 0 substituted; triethyl benzoyl or benzoyl benzoyl “JE as carylacyl M immunosuppressant; Aryl acyl substituted; benzyl or benzyl benzyl cmonomethoxyltrityl or monomethoxytril trityl dimethyl-t-dimethyl-tert-butylsilyl JB eg trialkylsilyl 5-R - or diphenyl methyl silyl 010116071083181171. Compound 71 may optionally be protected by, for example, trimethyl silyl dil; Trimethyl groups "Jilin" with a group of cytosine and these groups may be removed in a traditional way. The leaving group, 1, is an ideal leaving group, chlorine, such as chlorine, halogen, for example, nucleoside, in relation to groups known in the field of chemistry, such as acyl, ethoxy, or ethoxy, methoxy, Jie alkoxy, or bromine. or bromine (benzoyl sl acetyl chloroethane ET). The reaction in method A may be carried out in an organic solvent eg Lewis acid in the presence of acetonitrile or konitrile. 1,2-dichloroethane 00 trimethylsilyl triflate or stannic chloride 2-hydroxyacetaldehyde may obtain compounds of formula 7-a from = hydroxyacetaldehyde

Cr is protected in a suitable way for it

R,OCH,CHO Page ch Folder Can. 1. Research as described in lef magazine where 18 Y is known. The FAYOYT reaction results in 197 and full description of European Patent No. 4 where HSCH,CHOR, mercaptoacetal compounds according to the formula ¥ with mercapt and acetal. Known as HSCH,CHOC, Hs, as carbon containing -1; alkoxy atoms in which R is compounds of formula 1a where 0907 pp. 974-77; General AS Folder “Chem. Ber the field of that; You may transform Yay ethoxy or methoxy “Jil” for example 0 alkoxy OR in it, so be vo

Lad where IY to compounds of formula calkoxy L is compounds of formula "a where in it: tYo

11 By methods known in the chemistry of acyl or halogen carbohydrate sugars, the carbohydrate is oxo-isopropylidene glycerol -1,7 Ga. Compounds of the formula may be prepared, for example; Ry silyl by introducing 1,2-O-isopropylidene glycerol and removing the trityl isopropylidene group or benzyl trisubstituted silyl: acetic or formic acetic using a medium-strength acid such as isopropylidene formic acid Peroxide de gana alcohol is followed by oxidation of cacetonitrile in anhydrous zinc bromide or anhydrous zinc bromide. periodate iodate For method B; Substituted *-Fluoro 5-1000 may enter by known methods in

QS Art See 0

M.J. Robins, et al., in Nucleic Acid Chemistry, Part2, L.B. Townsend and R.S.

Tipson, editors, J. Wiley and sons, New York, 895-900 1978, and includes references such as; Article:

R. Duschinsky in Nucleic Acid Chemistry, Part 1, L.B. Townsend and R.S. Tipson, editors, J. Wiley and Sons, New York, 43-46 1978. | yo in trimethylhypofluorite and may be a fluorinating agent; For example; Trimethyl hypofluorite, fluorotrichloromethane, fluorotrichloromethane, and for method C, it is preferable to treat the compound represented by the formula CB 40701- trichlorophyllite chlorophosphate -4 aad i YL 1,24 tirazole azole dls to form compound 701 -4;- 4-chlorophenyl dichlorophosphate v. analogue which is then converted to compound 4-aminocytidine 441.2 4-triazolyl: .methanol desired by reaction with; for example; Methanol 4-aminocytidine by reacting the base (Jal as cay) The starting materials may be prepared according to formula B in a manner analogous to that described in method A. IY suitable optionally protected with a compound of formula and 0— S-fluorouracil Fluorouracil - o Vaccine Tajarisa ve. Aldrich Chemical Co., Milwaukee, 1 53233, USA. From S-fluorocytosine (yo)

VY : cist homologues and drans-homologs may be separated eg in a protected form by chromatography on silica gel 511108 with mixtures of organic solvents eg ethyl acetate/methanol Any protecting groups may be removed Using the appropriate reagent for each group. A° The compound represented by formula 1 may be converted to pharmaceutically acceptable amides s esters upon reaction with a suitable acylation agent; For example; halide or anhydride helps to acylate © 5-011 5 4-NH groups, hence; The acyl group may selectively be removed from one group or the other : from my group 5-011 5 4-NH for example; The treatment of the diacyl compound under acidic conditions, eg using Lewis acid, reduced zinc bromide 0: in methanol, removes the 4N-acyl group to obtain the corresponding 5-OH ester, as the treatment removes (J) i lo pg yl nd ALY A LS) p acyl ic seas sodium methoxide 5-011 to obtain the corresponding 4N-amide. WES acyl groups can also be removed by treatment with, for example, a commercially available esterase or ddipase enzymes; pig liver esterase 1 or pancreatic lipase; or by treatment according to Gok described in the full description of US Patent No. 071947 2. Compound J Beall in Formula 1 may be converted to its pharmaceutically acceptable salt in a conventional manner; For example; By treating with a suitable base. An ester or salt of a compound of formula 1 may be converted to the parent compound; for example (JO) by hydrolysis. A For a better understanding of the invention, the following examples are given by way of illustration. Method 1: cis-1-2-Hydroxymethyl-1,3-oxathiolan-5-yl-5-fluorocytosine A: Prepare: £-cis and *-trans-2-benzoyloxymethyl-5-N-acetyl-cytosin-1-yl-1,3-oxathiolane, and separate into trans cist analogues as described in the patent. ve European FAY No. chlor analogue cis-+ using trifluoromethyl hypofluorite in fluorotrichloromethane CCO3F and chloroform at -mother; bib of method 2,895-900, 1978 Robins, et al.

Nucleic Acid Chemistry, Part sass £8

: B -N, Ny-acetyl acetyl and ?- benzoyl lai 2-benzoyl Qasi Jira: Amsin dimethylamine in cethanol and the histolytic dissociates .%-cis-1-2-hydroxymethyl- 1,3-oxathiolan-5-yl-5-fluorocytosine Method B: Prepare: +-cis and +-trans-2-benzoyloxymethyl-5-uracil-1-yl-1,3-oxathiolane, > as Description in European Patent No. FAYOYT and after) 05 Protection of –Y – 2-hydroxyl hydroxyl group using a saturated solution of ammonia Li sal in cmethanolic Separation of the analogues on silica gel using BIOAC/MeOH European Patent No. 3,287077 as a flushing solution. The analogue 0 0 q is reacted with acetic anhydride in pyridine at room temperature to give “- acetate 2-acetate.” The solvent is removed in vacuum at 0 °C. At a temperature of 10 C. Then, 2-acetate |p is dissolved in CHCl and washed with aqueous sodium bicarbonate 201160115. The separated organic layer is dried, and CHCl is evaporated in vacuum. -0-AA5-07-At 777 -018-2-20607/107-+ CMSA description of Sel yo method A by the method of Robins et al. The process of converting the 5-F-uracil base into a base 5-F-cytosine by preparation of its derivative 4-triazol-1-yl 4-1,2 Tpq 1, 1171, 1984 B.Reese, J.

Chem.

Soc., © Perkins, and an article by ¢W.L.

Sung, Nucleic Acids Res. 9:6139, 1981 : using 1,2-4-triazole and two equivalents of -4-chlorophenyldichlorophosphate in dry pyridine at room temperature. x follows. - This conversion is a reaction with methanol previously saturated with ammonia at zero mm; And hydrolyzes 2-acetate to give +-cis-1-2-hydroxymethyl-1,3-oxathiolan-5-yl-5-fluorocytosine. Pharmaceutical combinations: Examples of the following combinations; The "active ingredient" is: +-cis-1-2-hydroxymethyl-1,3-oxathiolan-5-yl-5-fluorocytosine. disk combinations {Yo | . :

The following combinations f; b and c are prepared by means of a wet granulation process of the components 1 with a solution of povidone followed by the addition of magnesium stearate and pressing.

Yo. You. Active ingredient A 74 0 Lactose B.P. B 4 Yo Povidone B.P 1 00 C Sodium Starch Glycollate D E Magnesium stearate 5 and ove] ee] Combination B [eee]

You Yo. Active ingredient —- YY. Lactose b

YH Te Avicel PH 101 c q \o Povidone B.P d "1 0 Sodium Starch Glycollate e ¥ 5 Magnesium stearate and ve ee] the combination is 0 ee 0 active ingredient

You Lactose: 04 Starch

Povidone ¢ Magnesium stearate The following blends are prepared by direct pressing of the mixed ingredients. The milk sugar sO 1801088 in Blend E is of the direct-pressed type Dairy Crest-"Zeparox" Blend D 0 h/v

Yo. active ingredient

Yoo Pregelatinized Starch NF 15

\o . | º - combination — ‎emf] ‎Youll Active ingredient‎ ‎Vou | Lactose - Youu Avicel The blend and a blend with controlled release of the active ingredient The blend is prepared by wet granulation of the ingredients below with a povidone solution followed by the addition of magnesium stearate and pressing. : ede 00 a ous Active ingredient b 10 Hydroxypropylmethylcellulose Methocel K4M Premium c oy : Lactose B.P. d YA Povidone B.P e Magnesium stearate l Z > Release of the drug occurs during a period of about 4-7 hours; It will be completed after 11 hours. Example ? Blend A capsule mixes Prepare a capsule blend by mixing the ingredients for Blend D in Example ¥ above and filling them into a two-part hard gelatin capsule. Blend B below is prepared in a similar way. combination b aa fe a yo.

Active ingredient YET Lactose B.P. C Yo Sodium Starch Glycollate D Cy Magnesium stearate The combination is 1 mg/capsule Yo.

Active ingredient |

Macrogol 4000 B.P. and

The combination is dana [eke] Yo.

Active ingredient Yau Lecithin Yoo Arachis Oil Capsules containing Blend D are prepared by dispersing the active ingredient in ol lecithin and peanut oil and filling the dispersant into soft and flexible gelatin capsules. Combination E Active Ingredient Controlled Release Capsules: A combination active ingredient controlled release capsules is prepared by extruding the components a; b and c using an extruder; This is followed by sediment pelletizing and drying. The dried pellets were then covered with a controlled-release film and filled into a two-piece gelatin capsule. AA elf] Yo.

Active ingredient : : B \Yo Microcrystalline Cellulose C \Yo Lactose B.P. D VY Ethyl Cellulose Example ¢ Blend Gall AE 0 . Blend A Active ingredient: 0 | Hydrochloric acid solution, 0.1 M, or Sodium hydroxide solution, to 1 L M g.s. to pH 0.1 sterile water Sufficient up to 0 ml Dissolve the active ingredient in most of the water ©”**m-4m and adjust the pH between £0 sodium hydroxide § hydrochloric V,+ 5 0 as appropriate . Then the volume of the batch of materials is supplemented with water and filtered through a sterile filter in a sterile amber glass vial of 0 ml, type 1A, and it is closed tightly with stoppers and means to prevent leakage. . . 0 Blend With Live Active Ingredient Sterile, pyrogen-free, pH 7 phosphate buffer, Enough to So Je Yo

VY

© Example g Y Active ingredient g +1 Benzyl Alcohol g te Glycofurol 75 Is even enough? ml Benzyl Alcohol “benzyl alcohol” then add and dissolve glycofurol. The active ingredient in glycopherol is dissolved in microporous bay ales bottles and water is added up to ¥ ml. Then the mixture is filtered through a filter

Type A Ja 7 Sterilizer in amber color with a capacity of 1 0 eg syrup,; 8 g Active ingredient 8 g : Sorbitol Solution g ¥ Glycerol 1,000 g Sodium Benzoate eV Yo Flavor, Peach 17.42.3169

Jeo is enough for even Purified Water and most pure water. Then an aqueous solution of glycerol dissolving the active ingredient in a mixture of is added to the solution; followed by the addition of 80:01101 solution; And finally, sodium benzoate and the smell. Then supplement the volume with pure water and mix well.

You active ingredient

VY. Hard Fat, B.P. Witepsol H15-Dynamic Nobel in steam pressure cooker at #80 max. The component Witepsol HIS 5/1 molten is sifted using a Jalal mesh and added to the molten base material with active Aa 7010 through a silverson sieve equipped with a cutting head; Until a smooth dispersion is reached and the remaining Olea Ve temperature is compressed into the suspension and stirred to ensure a mixture of Witepsol 1115 is mixed at 45 C and a mesh is added and Yo is stirred. congenial. The entire suspension is passed through a stainless steel sieve iyo:

A continuous one is allowed to be cooled to Ee and at a temperature of 81 °C to Fe, 7.17 g of the mixture is filled into plastic molds? ml suitable. The pods are allowed to cool to room temperature. Yo.

Active ingredient YA Anhydrate Dextrose Magnesium Stearate L 0 Mix the above ingredients directly and prepare the vaginal suppositories by direct squeezing of the resulting mixture. J 4 Z Antiviral activity against Hepatitis C virus 1131 The compound cis-1-2-hydroxymethyl-1,3-oxathiolan-5-yl-5-fluorocytosine was tested as 1. Described below 1 It has been shown that the cell strain produced Human hepatitis C virus from HepGy 2.2.15 was described and characterized by Sells et al. See: (NAS 84: 1005, 1987 and J.

Virol. 62:2836, 1988 | It shares many of the characteristics of liver cells infected with chronic hepatitis C, and this virus is infectious, as demonstrated by the ability of V. 1 virus to cause disease in chimpanzees. This type of cell was used to test it outside the living animal to identify compounds with anti-HCV activity. Test compounds were treated to measure antiviral activity; Monolayer cultures were treated with the test compound at a concentration of 0-7000 micromolecular for ten days. DNA microarrays were harvested on day 3 from suspension fluid media containing DNA; the sixth and tenth; ve was treated with protinase K 1 mg/ml and 1 sodium dodecyl sulfate and incubated at +20 for one hour. DNA was extracted with equal volumes of phenol, followed by extraction with chloroform, and precipitated with ammonium acetate and propanol.

Schuell s Schleicher using the nitrocellulose method, DNA was collected on nitrocellulose and treated as S & S, 10 Optical Ave., Keene, NH 03431, Publication No. 700, 1987 Cells were collected and occurred Southern, J. Mol. Biol. 98:503, 1975. Cells described between cells after dissolution of cells with guanidine isothiocyanate DNA on the DNA found between cells in the same way as DNA. ammonium acetate and after deposition with LNA located outside

Hind II < intracellular nuclear endonuclease and oxy-acid endonuclease were inhibited for photo-quantification. Southern was then treated as described with agarose gel used on agarose gel 001 proliferative medium. Determine the effect of the antiviral compound by measuring a reduction of 0 times the amount of Dan particles; alii] given in HBV on the production of hepatitis C virus cis-1-2-Hydroxymethyl-1,3 -oxathiolan-5-yl-5-fluorocytosine Effect of Y=Y-10 buy cell primer in B covid-19 virus, integrated single-origin intermediate DNA sled complex, **Pg/ ml Agriculture sia Micromolecular Gram Today | hum | Today asl Sixth Tenth Salad Sua vv iy 1v oA AN 7 1,0 cells i

AA Yoo BE Ad vy YY A Untreated 0 y 111 nt Y CA 1,4 Yoo 0 Y 14 Ye 1 1,4 1,0 to 1 AY 31 10 VY 1,4 B m cells

AY As VY. q. 47 YY 1 1 unprocessed zero zero 4 q. 1 VA 1 zero zero Ye vi 1 : LY 1 You An hour from the tenth day of treatment. YE After Dane inside the cell HBV DNA particles were analyzed * +) Aplus all was an interruption (HBV DNA) 'Zero' indicates an undetectable level of ** milliliters. fpg 0 7 1

Claims (1)

0 Protectants or salt of cis-1-(2-(hydroxymethyl)-1,3-oxathiolan-5-y1)-5-fluorocytosine Use 1-1: of it for treatment or prevention of ester of a pharmaceutically acceptable salt or ester amide ester amide csalt Y B hepatitis virus infection r: : where 1?- use of the compound from protective element 1 cis-1-(2-( hydroxymethyl)-1,3-oxathiolan-5-yl)-5-fluorocytosine, 1 - of which is a pharmaceutically acceptable ester enantiomer or salt of the ester “amide or salt of the ¥ .enantiomer ¢ of an element protection? Where: 0S pall use = F 1 cis-1-(2-(hydroxymethyl)-1,3-oxathiolan-5-yl)-5-fluorocytosine, Y - of which the isolated ester enantiomer is pharmaceutically acceptable or a salt of ester camide or a salt of 1 .enantiomer ¢ where 1 ?- using the compound of protection 1 cis-1-(2-(hydroxymethyl)-1,3-oxathiolan-5-yl)- 5-fluorocytosine, Y of which is + a pharmaceutically acceptable ester enantiomer or salt of ester camide or salt; 7 : | + enantiomer ¢ Use of the compound in accordance with any of the preceding claims where said drug is in the form of # - 1 Dosage Unit. Y to 01 Use of the compound according to Claim © where said Dosage Unit contains 1 - 1 mg of compound: 0 Y cis-1-(2-(hydroxymethyl)-1,3-oxathiolan-5-y1)-5-fluorocytosine, ¥ of it. Pharmacologically acceptable ester or ester-camide salt “le or 1 Use of the compound according to claim 0 or 1 where the dose unit is in the form of a tablet or 7-1 capsule. Y ¢Yo