TWI198092B - Nucleosides - Google Patents

Abstract

Compounds of the formula I, wherein: R is independently H or -CH3 and pharmaceutically acceptable salts thereof have favourable pharmacological properties and are antivirally active against HBV and HIV.

Description

5. Description of the invention (1): It is related to the inhibition of nucleoside analogues, such as antiviral drugs, including reverse transcriptase and hepatitis B virus (HBV) DNA polymerase. The present invention proposes novel compounds with beneficial pharmacological variables, a pharmaceutical composition containing these compounds and a method of using HBV and HIV to inhibit virus and tumor-related diseases. International Patent Case No. W0 8 8/0 0 0 5 0 describes a series of 3, -fluorinated nucleotides, antiretrovirus and anti-HBV activity, including compound 2, 3, -dideoxy '3 —Fluoroguanine nucleoside (FLG) and 3, -fluorothymidine (FLT). This compound is clinically evaluated as an anti-ΗIV agent, and despite its excellent antiviral activity and pharmacokinetics, it still shows unpredictable toxicity (Flexner et a1, J "n-f Dis 170 (6) 13 9 4-40 3— (1 9 94)). The former compound, FLG, was extremely active in a test tube, but the inventors found that its bioavailability difference was about 4%-so the in vivo availability of the compound was limited to the animal model for intra-abdominal or subcutaneous administration. U.S. Patent No. 4, 963, 662. In general, 0, ^ π β buckle α α a series of 3, -fluorinated ribose and phase bis-carboxylate, and particularly >铷 Manufactured by Xi Xi, ^ LL will describe the FLT 5-0-brown fluorenyl derivative, but no improvement has been reported in the bioavailability 筚 π + Shangya. The international patent case W 0 9 3 1 3 7 7 8 describes ΑΑ β ft ft r π bucket from ^ 3 之 at the base 6 a position modified FLG and organisms, especially with positive-, A ^ nil ·- a -ν πί-ί * ^ Propyl-based,% butoxy, modified by cyproteron. The international patent 荦 Ν 0 9 3 1 4 1 03 describes the FLG derivative, which is produced by the hemp ΰ I do not look for it 00 ·… a λψ a々β Di Shi ', the forma is in the 6-position Oxygen is replaced by hydrazine, tritium, _ or% acid salt. 1198092 V. Description of the invention (3) The present invention also extends to the therapeutic use of compounds of formula I or salts, for example, pharmaceuticals can be made to treat retrovirus or TBV infection. In the treatment of conditions caused by a retrovirus (such as ΗIV), or HBV, the compound or salt of formula I is preferably administered in a dose of 50 to 1 500 mg, one, two or three times at a time, especially Is 100 -7 00 mg two or three times a day. It is hoped that a serum level of the active metabolite of 0.01 to 100 micrograms / ml can be achieved, especially 0.1 to 5 micrograms / ml. Therefore preferred compounds of formula I include: 5 '-0-[(S, R) 2, 3-bis- (L-valaminefluorenyloxy) -propanyl] -2', 3'-dideoxy -3'-Fluorine bird sigma rib ribs, and preferably 5'-0-[(8,10,2,3-bis-([-isoleukinoxy) -propanyl] -2 ' , 3'-dideoxy-3'-fluoroguanine nucleoside, 'and preferably 5 -0-[(R) 2,3_bis- (L-damineuroxy) -propionic acid] -2,3 -dideoxy-3'-fluoroguanine nucleoside, 5'-0-[(R) 2,3 -bis- (L-isoleucine fluorenyloxy) -propionamidine] -2 ', 3'-dideoxy-3'-fluoroguanosine; and pharmaceutically acceptable salts thereof. The compounds of the present invention can form salts, which constitute a further aspect of the present invention. Suitable compounds of the formula I are pharmaceutically acceptable Acceptable salts include salts of organic acids, especially carboxylic acids, including but not limited to the following: acetate, trifluoroacetate, lactate, gluconate, citrate, tartrate, maleate , Malate, steam salt, 2-hydroxyethanesulfonate, adipate, alginic acid

1198092

V. Description of the invention (4) Salt, aspartate, benzoate, butyrate, digluconate, cyclopentane; acid salt, glucosylheptanoate, glyceryl phosphate, oxalate , Heptanoate, hexate, fumarate, nicotinate, palmitate, pectate ester, 3-phenylpropionate 'bitterate' trimethyl acetate, Propionate, tartrate: acid salt, lactate salt, pivalic acid salt, camphor salt, alkanoic acid and succinic acid salt, organic luteolinic acid such as mesitolite, ethidium Sulphate, 2-sense ^ ^ | sulfonate, camphor sulfonate, 2 ~ naphthalene sulfonate, benzene sulfonate, p- ~ | Phenylbenzenesulfonate and p-benzenesulfonate ; And inorganic acids such as hydrochloride, hydrobromide, hydrosulfide, sulfate, bisulfate, hemisulfate,

I-cyanate, persulfate, linoleic acid and sulfonic acid. In some cases, the compound can be isolated in a hydrated state. x is a general practice for maintaining retroviral and HBV inhibitors, and is more interesting! It is to administer one to three additional island antiviral agents at the same time, such as AZZ \ ddl, ddC, d4T, 3TC, H2G, abacavir, ABT 6 0 6, j I f oscar net, via urea, ritonavir, indinavir, saquinavir, nevirapine, delaviridine, MIV 150 I efavirenz, Vertex 478 / amprenavir or Agouron AG1343, etc., used in the case of HIV, or lamivudine, interferon, I famciclovir, adefovir, lobucovir, BMS 200475 I L-FMAU, FTC, DAPD, Nabi 3700001, etc., are used in HBV examples. Such additional antiviral agents are usually administered in doses that can broadly reflect each other's therapeutic value. Relative to the compound or salt of formula I, a molar concentration ratio of mol: 00 to mol, especially 25: 1 to 1:25 is generally appropriate. Administration of additional antiviral agents for antiviral nucleosides intended for rash

Page 8 1198092 V. Description of Invention (5): When dyeing, it is usually less common. Although it is possible for the active substance to be administered alone, it is preferably presented as part of a pharmaceutical blend. Such a blend may contain an active agent as defined above, plus one or more acceptable carriers / excipients, and other therapeutic components as required. By definition of compatibility with other ingredients of the blend, the carrier must be acceptable and not harmful to the recipient. Concoctions include transrectal, nasal, topical (including cheek and sublingual),

! Vaginal or parenteral (including subcutaneous, intramuscular, intravenous and intradermal) administration, but the blend is preferably an oral blend. The blend may suitably be in unit dosage form I, such as lozenges and sustained release capsules, and may be prepared by any method well known in the pharmaceutical arts. This method includes the steps of combining the above active agent with a carrier. Generally speaking, the preparation of the blend is to uniformly and tightly mix the active component with a liquid carrier or a finely divided solid carrier or both, and then if necessary, shape the product. !

I The present invention extends to a method for preparing a pharmaceutical blend, which method comprises combining or combining a compound of Formula I: or a pharmaceutically acceptable salt thereof with a pharmaceutically acceptable carrier or vehicle. If the preparation of pharmaceutical blends involves intimate mixing of isochronous excipients with active ingredients in the form of salts, it is preferred to use excipients which are essentially non-alkaline, that is, acidic or neutral. The preparations for oral administration in the present invention can be in the form of separate units, such as capsules, cachets, or lozenges, each containing a predetermined dose of the active substance; powder or granules; active substance in an aqueous liquid Or non-aqueous liquid; liquid or suspension; or oil-water liquid emulsion or water-oil liquid emulsion, which is in the form of rapid and rapid injection.

1198092 V. Description of the invention (6)! For oral compositions (such as lozenges and capsules), suitable carriers include vehicles such as general excipients, such as binding agents such as syrup, acacia, gelatin, and sorbitol. , Tragacanth gum, polyethylene D to σ, each ketone (Povidone), ethyl cellulose, ethyl cellulose, sodium carboxymethyl cellulose, hydroxypropyl methyl cellulose, sucrose and starch ; Fillers and carriers, such as corn starch, gelatin,

Page 10 1198092 V. Description of the invention (7) includes an activated acid of the formula 〇

%

Fermented nuclear spine FLG

PGR — 0 PGR — 0 where PGR is R as defined above and protected with traditional protecting groups such as Fmoc, BOC or CBz N-. Activated derivatives used in tritiation may contain, for example, a fluorenyl halide, an acid anhydride, an activated acid ester, or an acid in the presence of a coupling agent, such as dicyclohexyl-disidized diimine. Representative activated acid derivatives include pro-based gas, anhydrides derived from alkoxycarbonyl halides such as isobutoxycarbonyl gas, etc., esters derived from N-hydroxy @ polamine, and esters derived from N-hydroxyphthalamide , N-hydroxy-5 -orthodifene-2,3-diamine, vinegar derived from 2,4,5-trisgas. Further activated acids are included in the formula RX, X represents the OR 'moiety, where R is

Page 11 1198092 V. R as defined in the description of the invention (8), and R 'is, for example, COCH3, COCH2CH3 or COCF3, or wherein X is benzotrisigma. The activated acid can be formed in advance, or it can be generated in the original using reagents, such as dicyclohexyl-imidized diimine (DCC) or 0- (1Η_benzotrisialyl-I-yl) N, N, N ', Ν' -Tetramethyl tungsten tetrafluoroborate (TBTU). When using a fluorenyl halide, such as a fluorenyl gas, a tertiary amine catalyst, such as triethylamine, Ν, Ν'-dimethylaniline, pyridine, or dimethylaminopyridine, can be added to the reaction mixture to release The hydrohalic acid.

The reaction is preferably carried out in a non-reactive solvent, such as N, N-dimethylformamide, tetrahydrofuran, dioxane, acetonitrile or a halogenated hydrocarbon such as digasmethane. If desired, any of the above-mentioned tertiary amine catalysts can be used as a solvent and care should be taken to allow for a moderate excess. The reaction temperature usually varies from 0 ° C to 60 ° C, but is usually maintained between 5 ° and 50 ° C. After 1 to 60 hours, the reaction is usually fully completed. The reaction can be tracked using thin layer chromatography (TLC) and a suitable solvent system. In general, when the reaction is known to be complete by TLC, the product is extracted with an organic solvent and purified by chromatography and / or recrystallization from a suitable solvent system.

Among them, donkey reaction occurs as a by-product of nucleoside bases, which can be separated by chromatography. However, such false deuteration can be minimized by controlling the reaction conditions. Such controlled conditions can be achieved by, for example, operating the reagent concentration or addition rate, especially the chelating agent, lowering the temperature, or the choice of solvent. Responses were controlled by TLC. The conventional 6-benzyl and especially 2 amine groups on the test group can be conveniently protected with traditional protecting groups to prevent erroneous acidification.

Page 121198092 V. Description of the invention (9) The preparation of 2,3-bis-N-protected amino fluorenyl glycerol intermediates is based on the traditional protective groups, such as oxybenzyl to protect glyceric acid. Carboxyl and esterified with a suitable N-protected amino group, and then selectively deprotected methoxybenzyl. R2 and the amino acid derivative, if present, can be further esterified to a linker group with 2-fluoren-4-acyl-cycloalkane-1,3-dione, the method is described in International Patent No. 1 W0 94/29311, the contents of which have been listed for reference in this case. ! Detailed description of Example 1! 2, 3 -Bis (N —CBz-L—Polysulfonyloxy) -propion I-man (MSS-137) i | 〇 | Α〇η | _ N-CBz-Valaminofluorenyl-1-Ia! Ia) 2, 3- (bisN-CBZ-L-Valaminofluorenyloxy) propionic acid 1,1-dimethylethyl Ester I p-1,2-dihydroxypropionic acid 1,1-dimethylethyl ester (2.43 g, 15 mmol), N-CBZ-L-valinic acid (7.54 g, 30 mmol) Moore) and 0th birthday? (0.37 | g, 3 mmol) to 150 ml of a solution of digas methane, DCC (7.2 I g, 35 mmol) was added, and the mixture was stirred at room temperature for two days. The filtrate was evaporated, ethyl acetate was added, and the organic phase was washed twice with 5% acetic acid, 5% sodium bicarbonate, and water. The organic phase was dried over sodium sulfate and filtered and evaporated under reduced pressure. The product was separated by silica gel column chromatography. Yield: 8.2 g = 86%. ! lH-NMR (DMSO d-6) 0.87 (m, 12H) 1.40 (d, 9H) 2.12 I (in, 1H) 4.0 2-4.4 0 (m, 2H) 5.04 (d, 4H) 5.20 (m, I 1H), 7.36 (m, 10H) 7. 72 (d, 2H)

Page 131198092 V. Description of the invention (ίο) b) 2, 3-bis- (N-CBZ-L-valamineamidooxy) propionic acid p-ester (7.2 ^ -NMR 3.92-4 5.28 (Example 2 2,2,3,-3 -bis- (N-CBZ-L-valamine methoxy) propionic acid 1,1-difluorenylethyl g, 11.4 mmol Mol) in a solution of dioxane (25 ml), fluoroacetic acid (25 ml) and the solution was stirred at room temperature for 5 hours. The solution was evaporated and co-evaporated with toluene twice. The product was packed in a silica gel column Layer. Yield: 5.9 g = 90%. (DMSO d-6) 0.92 (m, 12H) 2. 08 (d, 2H) .17 (m, 2H) 4.3 0 -4.67 (m, 2H ) 5.04 (s, 4H) m, 1H), 7. 32 (m, 1 OH) 7. 70 (d, 2H) dideoxy-3 '-fluoro-5' -0- "2, 3 -bis -(L-valaminyloxy) propionyl 1 uridine nucleoside a) 2,, 3, deoxy-3 '-fluoro-5' -0- [2, 3 -bis- (N- CBZ-: L-valaminyloxy) propionyl] guanine nucleoside (MSS-138) 2 ', 3'-dideoxy-3'-fluoroguanine nucleoside (2.15 g, 8 millimoles), 2, 3 -bis- (N-CBZ-L-valaminemethyloxy) propionic acid (6.2 g, 10.8 millimoles), DMAP (244 millimoles , 2 mmol) and Β0BT (1.46 g, 10.8 mmol) were co-evaporated with DMF twice and reduced to about 120 ml. DCC (2.48 g, 12 mmol) was added Mol) and the mixture was stirred at room temperature for two days. The mixture was filtered and the solution was evaporated under reduced pressure. 150 ml of ethyl acetate was added and the organic phase was washed twice with 5% acetic acid, 5% sodium bicarbonate and water. The organic phase was dried over sodium sulfate and evaporated under reduced pressure. The product was separated by silica gel column chromatography, yield = 2.25 g = 35%. ^ -NMR (DMSO d-6) 0.88 (m, 12H ), 2.12 (m, 2H),

Page 141198092 V. Description of the Invention (11) 2.50-3.00 (m, 2H), 3.88-4.14 (m, 2H), 4.22-4.62 (in, 6H), 5.04 (s, 4H), 5.30-5.61 (m, 2H), 6.16 (m, 1H), 6.50 (s, 2H), 7.32 (in, 10H), 7.70 (in, 2H), 7.92 (s, 1H) b) 2 ', 3'- Dideoxy-3'-fluoro-5'-〇- [2,3-bis-([-valamineamidooxy) propionamidine] guanine nucleoside 2,3,3, _fluoro-5 , -0- [2, 3 -Bis- (N-CBZ-L-Valaminomethyloxy) propanyl] guanine nucleoside (0.41 g, 0.5 mmol) in ethyl acetate A solution of the ester (40 ml) and acetic acid (20 ml) was hydrogenated with palladium black (200 mg) at 30 psi for 2 hours at room temperature. The catalyst was filtered and washed with ethyl acetate and acetic acid. The solution was evaporated under reduced pressure and the product was dried under vacuum to form a dihydrochloride salt. The slug rate = 0.3 g = 95%. l-NMR (DMSO d-6 & D20) 0.9 4 (m, 12H), 2.18 (m, 2H), 2.5 2-3.0 0 (in, 2H), 3.8 8-4.0 9 (m, 2H), 4.36- 4.72 (m, 6H), 5. 4 2-5. 72. Biological Example 1 Pharmacokinetics Confirmation that FLG can be released in vivo by the oral prodrug of the present invention in rat mode has been confirmed to be a useful model for assessing pharmacokinetic variables of nucleoside analogs. The oral composition was administered in a pharmaceutical vehicle, including propylene glycol, to a triple fasted animal at a dose equivalent to 0.1 mmol / kg. For comparison, another group of rats was iv. Administered a metabolite of 2 ', 3'-didehydro-3'-fluoroguanine nucleoside at 0.01 mol / kg. Collected at 0.5 to 6 hours after administration (5 minutes to 6 hours at F LG) at regular intervals

Page 15 1198092 V. Description of the invention (12) Serum levels of metabolites in the serum of individual animals. Metabolites were analyzed by HPLC, 254 nm UV detection in a manner similar to that described by Stahle et al 1995, J. Pharm. Biomed. Anal. 13, 3 6 9-37 6. The HPLC system was buffered with 0.05M ammonium dihydrogen phosphate buffer solution, plus 1.2% 2-propanol solvent, to pH 4.5, or 30mM sodium dihydrogen tank acid, and 2% acetonitrile solvent, buffered to Based on pH 7.0. The column can be 100 x 2.1 mm BAS C18 5 micron particle size with a guard column of 7 micron C18, or Zorbax SB-CN C18 150x4.6 mm, 5 micron column. The protein binding of the compounds of the present invention is negligible, just like metabolites ' and is useful for ultrafiltration on Microcon 30 or Amicon filter membranes for corporate clear samples. Beneficially, subjecting the main peak to further column chromatography will help FLG go beyond the resolution of low molecular weight serum components. The iv level can be increased up to 10 times to get the AUC value and compare it with the oral level. The absolute oral bioavailability can be determined by the ratio between g-AUCiv and 0_AUC. The compound of Example 2 showed an absolute bioavailability of 68% '72% and 64% of 0-6 hours, resulting in plasma levels of active metabolites sufficiently higher than antiviral inhibitory levels, as reported in the scientific literature of. Therefore, the compound of the present invention significantly enhances the oral bioavailability compared with the metabolite 2,3, -dideoxy-3, -fluoroguanine nucleoside. Significantly, the compounds are released into the bloodstream in a fairly continuous manner, rather than in an immediate peak pattern. This means that the effective dose of the active substance can be used in the bloodstream for several hours, which helps the daily dose at one time. In addition, continuous release can prevent the compound from being seen at a faster release rate = two toxicity issues. , Blending example 1

1198092: V. Description of the invention (13): The following components of the tablet blend are sieved through a 0.1 mm sieve and dried and mixed; 10 grams of 5'-0-[(R) -2,3 -double- ( L-Valamine 醯 oxy) propionamidine]-! 2 ', 3'-dideoxy-3'-fluoroguanine nucleoside! 40 grams of lactose | 49 grams of crystalline cellulose! 1 grams of stearin Magnesium acid:! Use a tablet mill to compress the mixture to a tablet containing 250 mg of active ingredient |

I | Blends Example 2! Casing tablets 丨 The tablets of Blend Example 1 are sprayed in the tablet coating machine with the following solution |-! Coating, the solution contains ~ 120 g of ethyl cellulose and 30 g of propylene glycol! ίο sorbitan monooleate

I was added to 1000 ml with distilled water. I Blend Example 3!

; Controlled release blend I 50 g 5'-0-[(R) -2,3-bis- (L-valaminepyroxy) propanyl] I-2, 3 -dideoxy- 3-Airbird σ-epirubin nucleus; 12 g of propyl fluorenyl cellulose (Methocell Κ15)

I. 4.5 grams of lactose! Mix dry and granulate with an aqueous paste of buprene. Add magnesium stearate

Page 17

1198092 V. Description of the invention (14) (0.5 g) and the mixture was compressed in a tablet mill to a 13 mm diameter tablet containing 500 mg of active substance. Blends Example 4 Soft Capsules 250 grams 5'-0-[(S) -2, 3-bis (L-valaminepyroxy) -propylpyridine] -2 ', 3'-dideoxy- 100 g of 3′-fluoroguanine nucleoside 100 g of peanut oil and filling it with soft gelatin The compound of the present invention is dispersed in capsules of egg filling and peanut oil. m

Page 18

Claims (1)

1198092 Case No. 87113427A01 Year 0 S || :: .. 6. Scope of patent application 1. A compound V of formula I. —-Pr r 〇 NH N ° ~ 1 ^ ^ N NK F Among them: both R groups are H or both -CH3 and pharmaceutically acceptable salts thereof. 2. The compound according to item 1 of the patent application, wherein both R groups are hydrogen. 3. —An anti-HI V or anti-HBV pharmaceutical composition, which contains a compound as defined in item 1 or 2 of the patent application scope, and a pharmaceutically acceptable carrier thereof. The compound of item 2 is used for the preparation of pharmaceuticals for treating HIV or HBV. O: \ 57 \ 57208-930527.ptc Page 20