DE2162616A1 - l- (2- or 3-Deoxy-beta-D-pentofuranosyl) -2-thiouracile and 1-beta-D-pentofuranosyl-2-thio-6-aza-uracile, process for their preparation and pharmaceutical preparations containing these compounds contain - Google Patents

Description

2152616

LAWYERS -

^ JÜ2. DlPL-CHEM. WALTER BEIT Dec. 15, 1971

■ IFRSD HQ = P? EiiER
.1. JUR. ÜifL-CH-iM. H.-J. WOLFP
R. JUR. HOUSE CHU. AX

i-i3 FRANKFUKTAM MAIN-HOCHSf

ADaONS18ASS65i

Our Kr. 17 53o

■ The Upjohn Company
Kalamazoo, kich., V.bt.A.

l- (2- or 3-Deoxy-ß-D-pentofuranosyl) -2-thiouracile and l-ß-D-pentofuranosyl-2-thio-6-aza-uracile, process for their preparation and pharmaceutical preparations containing these compounds contain.

The present invention relates to new 1- (2- or 3-i> eoxy-fi-D-pentofuranosyl) -2-thiouracils and 1-β-li-eentofuranosyl-2-thio-6-azauracils, and a process for their Production which is characterized in that a corresponding 2-u-i-aethylurac <i> l-rru.cleoside or 2-u-methyl-6-aza-uracil nucleoid is treated with uchwei'olwaüserstoi'f. The new compounds are effective against viruses and neoplastic cells, and the present invention also includes preparations containing the new compounds.

liie novel 1- (2- or 3-i> eoxy-ß-jj-pentoiura-

? Π982Π / 1156

nosyl) -2-thiouracile and i-ß-lJ-pentofuranosyl ^ -thio-e-azauracile "have the following formula

HN
S.

YGH

CM

i I

in which M is hydrogen, a lower alkyl radical with 1 "to 4 carbon atoms, a hydroxy-lower alkyl radical with 1" to carbon atoms, a lower alkoxymethyl radical with 2 "to 5 carbon atoms or a lower alkanecarboxylic acid ester of the hydroxy-lower alkyl radical with a total of 5 up to 6 carbon atoms, X the grouping = GH- or = N-, Y a hydroxyl group, a lower alkanoyl radical with 1 to 4 carbon atoms or an aroyl radical with 7 to 8 carbon atoms and Z, Z ^a , Z, Z ^c independently of one another Represent a hydrogen atom, a hydroxyl group, a lower alkanoyl radical with 1 to 4 carbon atoms or an aroyl radical with 7 to b carbon atoms, with the general assumption that at least one of the groups, but not more than two simultaneously hydroxyl,

2 0 9 8 2 9/1 1 5 B.

are lower-alkanoyl or aroyl groups, with the further proviso that Z ⁰ and Z ^c , Z ^a and Z ^c , Z and Z ^a and Z and Z ^b cannot both be hydroxyl groups or lower alkanoyl groups when X is a = CH -G group is.

liie chemotherapeutically active compounds are those of the formula I and i-ß-D-kibofuranosyl ^ -thiouracil-Iierivate, in which Z ^a and Z denote hydroxyl, lower alkanoyl or aroyl.

The substituent M in the above formula I can be a methyl, ethyl, Represent propyl or butyl group or isomeric forms thereof. Furthermore, M can be a hydroxymethyl, 2-hydroxyethyl, 3-wydroxypropyl, 1-Hydroxyäthyl- or 2-Hydroxypropylrest continue to be a wethoxymethyl, ethoxymethyl, propoxymethyl, isopropoxymethyl or butoxymethyl radical, or for example an acetoxymethyl, propionoxymethyl, 2-butyroxyethyl radical or the like

If the substituents Z, Z ^a , Z, Z ^c and Y are lower alkanoyl radicals, they can consist, for example, of the acetyl radical (preferred), a propionyl, butyryl radical or isomeric forms thereof. Possible aroyl radicals for the substituents mentioned are, for example, the benzoyl, loluoyl or anisoyl radical.

The ß-D-Pentoffuranosylzucker with X = = 1ϊ- include "for example Kibose, arabinose, xylose and lyxose and their deoxy forms.

never effective against \ Tiren and tumors according to the invention 1 -Jä-i) -Ientofuranosyl-2-thiouracile and 1-β-'JJ-pentofuranosyl-2-thio-6-azauracile are made using an improved direct synthesis with hydrogen sulfide. According to In a preferred embodiment, a solution is a 1 -ß-u-Pentofuranosyl-2-ü-methy! Uracils or 1 -ß-'Jj-Pentofurano-

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syl-2-ü-methyl-6-aza-uracils with gaseous hydrogen sulfide Treated at a temperature between 15 and 30 ° C in the presence of a basic catalyst until the 2-0-kethyl group is replaced by a sulfur atom. The desired product is easily obtained in relatively pure i'orm and generally in Maintain good yields, in a conventional manner by evaporating the solvent and chromatographic Cleaning up.

The 1-ß-D-pentofuranosyl-2-umethyluracils used as starting material and -2-ü-methyl-b-aza-uracils are in general known compounds, which are based on those of Brown, 'i'odd, and Yaradarajan, J. Ghemr Soc. 868 (1957) Methods are available.

According to a preferred embodiment of the invention, a 2,5'-anhydro-i-ß-u-pentofuranosyluracilj * or -6-aza-uracil reacted with methanol in the presence of an alkali metal alkoxide. According to the prior art, ammonia and triethylamine were used worked as basic catalysts; in some cases the conversions are uselessly slow here.

It is also possible to use an alcohol other than methanol, e.g. ethanol or propyl alcohol, as well as another effective alkali metal alkoxide, e.g. sodium methylate (preferred), Sodium ethylate, sodium propylate and potassium methylate. The xieaction runs smoothly at temperatures in the range of 25 C, but slightly higher temperatures of, for example, to can be applied to about 40 ° 0.

The desired 1-ß-D-pentofuranosyl-2-U-methyluracil or 1-ß-D-pentofuranosyl-2-ü-methyl-6-aza-uracil is worked up by neutralizing the reaction mixture in portions with acid, e.g. with a lower alkanoic acid such as iissetic acid (preferred) or propionic acid, followed by the solvent

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removed and a conventional cleaning system is connected ', e.g. by chromatography, recrystallization or the like.

The compounds used as starting materials 2,5'-anhydro-1-ß-u-pentofuranosyl-uracil and -6-aza-uracil are known or can be used according to the literature known ketiiodes or the the following regulations.

The activity of the compounds according to the invention against viruses and neoplastic cells were detected both in vitro and in vivo. Preliminary experiments with tissue cultures made from L-121o leukemia cells showed that 1-ß-D-i-i.ibofuranosyl-2-thio-6-aza-uraeil in very low concentrations

v / is ineffective. The ID [- "is, for example, o, o6yug / ml and ^{the IJ)} Sü ^o » 23 /

The following experiment was carried out in vivo:

'/ stabs (käuse) received 1 x. 1o ascitic tumor cells (L 121o), and the compound 1-du-kiboruanosyl-2-thio-6-aza-uracil v / urde in an amount of 200 mg / kg per l'ag intrapex-itoneal for 8 days administered. In the treated houses, the survival time was 47% longer than the survival time in untreated lice after infection.

These observations result in a daily dose for mammals from e tv / a 1 ο mg to around 4oo mg per kg of body weight, never exact jjosxs depends of course on the type of tieres, z.L. riuuLd, cat, xvindvieh, horse, mouse, iiatte or Human, in front of age, or worse of the disease state and the strength of the patient or animal. The administration can be drawn with daily i-lots, several doses per 'i'ag or taking place under i'.placing administration breaks »

Those according to the invention have proven to be extremely effective Connections in tests against viruses. At different

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Tests received the compounds 1-ß-l) -i \ .ibofuranosyl-2-thio-6-aza-uracil, i-ß-u-iiibofiiranosyl-2-thiouracil, 1- (2-leoxy-ß-I) erythroperito ± \ iranosyl) -2-thiothymine, 1- (5, 5- ±) i-U-acetyl-2-deoxy-JB-jj-ery LiiroperLtoi'uranosyl) -2-thiothymine, 1- (2-deoxy-ß-L) -erythropentoiuranosyl-2-thiouracil, 1- (3,5-I> i-0-acetyl-2-deoxy ß-ii-erytnropentofuranosyl) -2-thioiiracil and 1 - (2-.ueoxy-ß-l) -threopentoiuranosyl) -2-tniothyniin the ratings 5 "to 4+ (best value) with regard to their activity against nerpes viruses. The compounds are also effective against Goe virus and HA-1 virus.

preferred compounds of the present invention are the effective against viruses 1- (2- or 3-i) eoxy-ß-jj-erythropentofuranosyl) -2-thiouracils, in particular 1- (3,5-l »i-ü-acetyl-2-deoxy-β-i; -erythropentoxuranosyl) -2-thiothymine and 1- (3,5-iJi-0-acetyl-2-deoxy-β.-jj-erythropentofuranosyl) -2-thiouracil.

A few other preferred compounds are the 1-ß-i) -pentofuranosyl-2-thio-6-aza-uracils, in particular the 1 -ß-Li-itibofuranosyl-2-thio-6-aza-uracils, especially 1-ß- l) -iiiboxuranusyl-3 ^r -2-thio-6-aza-uracil and its 2 ', 3'»5'-triacetate.

i) the valuable anti-virus and anti-tumor effect of the invention 1-ß-I) - ± -entoruranosyl-2-thiouracile and -2-tnio-6-azauracile is exploited by bringing the virus to be combated or an aleoplasm into contact with the compounds. This is achieved by methods known per se of infusion into the environment containing the virus or the neoplastic Contains cells. For example, a syncytic virus can infect the respiratory tract in chimpanzees fight by 1- (3,5-ui-U-acetyl-2-deoxy-i.i-iJ-erythropentofuranosyl) -2-tniotJ: iymin or 1- (2-i) eoxy-β-jJ-threopentofuranosyl) -2-thiothymine on the mucous membranes of the nose and Throat applied in a concentration of about 1, ί. L'ie so applied to an infected or adjacent tissue

209B29 / 1186 ' ^BAD ° ^RIGINAL

Compound is distributed through the tissue fluids and thus comes into contact with the virus at the site of the infection. The treatment can also be carried out systemically by the "compounds in formulations for oral or parenteral Administration incorporated.

For oral administration, the active ingredient is combined with a pharmaceutical Carriers combined to form a solid or liquid dosage form.

Suitable solid dosage forms are tablets, pills, Capsules, granules, powders, suppositories and the like. Appropriate the pharmaceutical carriers used for these dosage forms are mostly starch, lactose, dicalcium phosphate, terra alba, («Eaf calcium sulfate), talc, stearic acid, magnesium te ar at or rubbers. Suitable liquid i'Orms are solutions, suspensions, Syrups and emulsions. Conveniently, the pharmaceutical carrier in such liquid dosage forms comprises water, Oils and water emulsions. If desired, suitable ones can be used Dispersing or suspending agents may be added, e.g. Tragacanth, gum acacia, alginates, dextran, sodium carboxymethyl cellulose, Methyl cellulose, polyvinyl pyrrolidone, gelatin and mixtures thereof. Suitable oils for ■ the production of solutions and water-oil emulsions are e.g. cottonseed oil, sesame oil, Coconut oil and .ord nut oil.

For parenteral administration, the 1-ß-D-pentofuranosyl-2-thiouracils and -Z-thio-ö-aza-uracile in the form of diluted ones aqueous solutions, aqueous suspensions and oily dispersions for intramuscular injection, drip infusion, vaücular Perfusion or the like. Be formulated. A solubilizer, e.g. N, ii-dimethylacetamide (preferred), is expedient. i <l, W-iemetiiylformamid, iithanol or the like. Used. If desired, other aqueous media such as injection water,

BATHROOM OFUGINAL 209829 / 11SB

2152616

normal saline solution, itinger solution, blood plasma or whole ± ilut uses v / earth.

Formulations according to the invention for topical use are e.g. powders (preferred), ointments, creams, pastes, free and the like. ■ Such preparations can contain emulsifiers, solvents, antioxidants, preservatives, buffers and extenders.

example 1 Production of 1-ß-D-Ki * boi'uranosyl-2-thip-6 ~ aza ~ uracil

Part A: 1- (2,3-0-Isopropylidene-β-1) -ribofuranosyl) -6-aza-uracil

With the exclusion of moisture, a suspension of 245 mg (1.0 millimoles) 2-ß-D-ribofuranosyl-as-tr / azin- (2M, 4H) -dione, 49o mg (3.8 millimoles) of anhydrous copper sulfate and 10 ml Acetone acidified with 0.1 ml of concentrated sulfuric acid and stirred continuously for about 24 hours. The suspension is then filtered and the filtrate is made more aqueous by the dropwise addition of 1.0 Sodium hydroxide solution adjusted to neutral pH-Vert. The neutralized i'iltrat is then "at reduced Pressure evaporated to dryness. The residue is dispersed in a mixture of chloroform and water and the organic Phase is separated from the aqueous phase. The aqueous phase is extracted several times with chloroform, the extracts are combined with the chloroform phase. .After drying the chloroform is evaporated from the chloroform solution under reduced pressure. This gives a white solid, the when recrystallized from a mixture of acetone and methylcyclohexane 62.5 mg of 1- (2,5-0-isopropylidene-ß-D-ribo: iuranosyl) -ö-aza-uracil from melting point 141.5 to 143 O results.

209829/1 1 56

Ultraviolet Absorption:

Λ ^; ^Äthan01 261 γ (έ = 7 οοο)

Cli characteristic. Lk-is / absorption bands (cm "") OH / IMJti: 353o; i> m / = GH / uri: 314o, 3oSo, 295ο; C = O / C = u; 1585 (weak); C-O / C-N / others: 1275, 12o5, 1165, 1135, 1115, 1o7o, 1o55; others: 87o.

Analysis: iier. for O ₁₁ H ₁₅ K ₅ U ₆ : C: 46.31; H: 5.3o; W: 14.73. Found: O: 46.31; H: 5.52; N: 14.96.

i'eil U: 1- (5-u-tosyl-2,3-u-isopropylidene-ß-.u-ribofuranosyl) - -6-aza-urac ± l.

With the exclusion of moisture, a solution of 2.58 g (13.5 i'Iillimol) purified p-toluenesulfonyl chloride and Mix 26 ml of pyridine to a solution of 4.39 g (approx. 10 Hillimol) 1- (2,3-0-isopropylidene-β-D-ribofuranosyl) -6-aza-uracil (For preparation see Section A) and 26 ml of pyridine, the had been cooled to -1o G, added dropwise. The temperature is kept between 0 and -1o C during the addition. The that Flasks containing p-toluenesulfonyl chloride are added to 4 ml of pyridine are rinsed out, and this rinsing solution is added to the reaction mixture admitted. The mixture is then left to stand at about 3 CJ for about 16 hours. The reaction mixture turns dark it is then added with a kicked J ^ is, then again left for several days at approx. 3 G. After the pyridine and water have been evaporated off under reduced pressure, the residue becomes Azeotropically distilled with absolute ethanol at reduced pressure, ner non-volatile residue v / ird in a mixture dispersed from 75 parts of chloroform and 15 parts of water. After separating the chloroform layer from the wäijsrif; en phase the former is washed with water. the Washed, dark chloroform layer is then placed over anhydrous Sodium sulfate dried and freed from .Chlor-üJO-an under reduced pressure. The residue is stored in hot abso-

BATH ORIGINAL

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- 1ο -

Dissolved in pure ethanol and treated with decolorizing charcoal. This is then filtered off and washed on the filter with absolute ethanol. According to a second decolorization with charcoal, the alcohol is evaporated under reduced pressure, lter thus obtained residue (yellow solid) is χ 2 at a reduced pressure azeotropic distillation using 95 / ^j aqueous ethanol. The residue obtained in this way is kept in vacuo in order to remove traces of pyridine. The pyridine coolant is then dissolved in acetone and decolorized carbon is added. This is filtered off and washed on the filter with acetone. The acetone filtrate is concentrated to about 75 ml and diluted with 50 ml of technical grade hexane (Skellysolve B, G-mixture of isomeric liexanes with a boiling point of 60 to 71 ° C.). The diluted solution is seeded with crystals and left to stand for about 16 hours at 25 G, then for several hours at 3 ° C. The crystals that form are collected on a filter, washed well with ethyl ether and dried under a vacuum at 25 O. 2.12 g of 1- (5-u-tosyl-2,3 - ^ - isopropylidenide-D are obtained in this way -ribofuranosyl) -6-aza-uracil from melting point 156 to 158 C.

Part C: 2,5'-Ahydro-1 - (2,3-ü-isopropylidene-i3-jj- £ "ibofuranosyl) -6-aza-uracil

a mixture of 300 mg (0.705 hillimol) 1- (5-0-'x'osyl-2,3-isopropylidene-β-D-ribofuranosyl) -6-aza-uracil (see Part B), 10 ml of dimethylformamide and 325 mg of sodium bicarbonate are heated to 150 ° C for 10 minutes with the exclusion of moisture in a nitrogen atmosphere, after the reaction mixture has cooled to 25 ° C, it is filtered and the filter is carefully washed with dimethylformamide, i- 'The filtrate and wash solutions are evaporated to the'i'x'ockene under reduced pressure. December ¹ Kückstand thus obtained is χ 2 with absolute Atnanol under reduced pressure, azeotropic distillation of a subject to obtain a yellow solid, jjieser J5 / öigem aqueous ethanol is dissolved in ^c, the solution is treated with 2.5 g of

BATH ORIGINAL 209829/1158

Silica gel added. The slurry is evaporated to Tx-ockene by evaporating the aqueous ethanol "under reduced pressure. The dried silica gel is applied to a column containing 28 g of silica gel which has been mixed with" solvent A "(mixture of ethyl acetate, technical hexane and methanol in the ratio 6: 3: 1). The desired product is eluted with further solvent A. After evaporation of the solvent A "under reduced pressure, 79.1 mg of residue is obtained, which is crystallized from a mixture of acetone and technical grade hexane . This gives 65.1 mg 2.51 ^! & nhydro-1- (2,3-0-isopropylidene-ß-D-ribofuranosyl) -6-azauracil, which has no defined melting point. Ultraviolet Absorption:

236; 26o (black group) m / u (t = 121oo; 595o)

Characteristic. Ik absorption bands (cm ") C = o / C = N: 1675, 1595; Ο-Θ / σ-Μ / others: 1285, 126o, 12o5, 1o95, 1o65; other 91 ο, 855.

• Analysis: Ber. for G ₁₁ H ₁₅ N ₂ O ₅ : C: 49.43; H: 4.90; N: 15.73. Found: C: 49.33; H: 5.16; N: 15.53.

Part Ό: 1- (2,3-0-Isopropylidene-ß-D-ribofuranosyl) -2-thio-6-aza-uracil

A solution of 68.4 mg (0.26 millimoles) of 2,5'-anhydro-1- (2,3 - ^ - isopylidene-β-D-ribofuranosyl) -6-aza-uracil (For preparation, see Part Ci), 2 ml of dimethylformamide and 3 drops Triethylamine is treated with gaseous hydrogen sulfide by slowly blowing the gas for half an hour lets pearl through the reaction mixture. The solution will then Concentrated to dryness at reduced pressure, the residue v / ird 2 with absolute ethanol at reduced pressure a

inafi subjected to azeotropic distillation, whereupon the 1- (2,3-0-isopropylidene-ß-iJ-ribofuranosyl) -2-thio-6-aza-uracil is obtained as an amorphous Ji'e substance.

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Part] ü: i-ß-D-nibofuranosyl ^ -thio-o-aza-uracil.

Koh.es 1- (2.3-0-isopropylidene-ß-D-ribofuranosyl) -2-thio-6-aza-uracil (prepared according to part D of 84.1 mg (o, 315 toillimol) 2.5 ^I -Anhydro-1- (2,3 - <- ⁾ -isopropylidene-ß-D-ri'bofuranosyl) -6-aza-uracil) was dissolved in 4 ml of a mixture of 9 parts trifluoroacetic acid and 1 part V / water, the The solution was left to stand for 25 minutes "at 25 ^° C. After the volatile components had been removed by evaporation" at reduced pressure, the residue obtained in this way was subjected to azeotropic distillation with 95% aqueous ethanol "at reduced pressure treated again with the 9: 1 mixture of trifluoroacetic acid and water for 30 minutes, then the reaction mixture was worked up as described above. The residue of the azeotrope treatment was dissolved in 1 ml of ethanol and the ethanolic solution was applied to a column with 2o g of silica gel, which was moistened with a 1: 1 mixture of solvents A and B (solvent B is a mixture of Ethyl acetate, technical hexane and methanol in a ratio of 8: 12: 1). The desired product was eluted with the same 1: 1 mixed solvent, collecting fractions of 1 ml each. Fractions 46 to 50 were combined and freed from the solvent under reduced pressure, the i-ß-D-kibofuranosyl-2-thio-6-aza-uracil being obtained.
Ultraviolet Absorption:
itOH ₂₁₉ . _{272) 5}

Example 2 Preparation of i-ß-D-ri.ibofuranosyl-2-thiouracil

Part A: 1- (2,3-ü-Isopropylidene-β-D-ribofuranosyl) -2-thiouracil

8 ^ 9/1156

A solution of 744 mg (2.5 millimoles) 1- (2 _tons 3- ⁽ -'-Isopropylidenß-D-ribofuranosyl) -2-0-methyluracil (prepared by the method of Brown, Todd, and Varadarajan, J. Chem Soc. (1957)), 9 ml of dimethylformamide and 0.45 ml of freshly distilled triethylamine were treated with hydrogen sulfide by bubbling a stream of gas through the solution for a gentle 20 hours. A green solution was obtained. Solvent and volatile constituents were evaporated off under reduced pressure and the residue thus obtained was subjected to azeotrope treatment 3 times with benzene at reduced pressure. This gave 85o mg of an amorphous yellow foam. This foam was dispersed in 6 ml of warm solvent A and the mixture for several days at 25 ° C was allowed to stand crystals formed, which were collected on a filter and washed first with the l ^{|. l} were washed iltrat and then with cold solvent a was filtrate and / aschlösungen 7 under reduced Dr evaporated to dryness, giving an amorphous residue. This was dissolved in chloroform, and silica gel was added to make a slurry. The chloroform was then evaporated and the dry absorbate of silica gel was applied to a column of silica gel wetted with solvent A. The column was then developed with solvent α, collecting fractions of 5 ml. Fractions 50 to 39% are combined and evaporated to dryness under reduced pressure. This gave 1- (2,3-u-isopropylidene-ß-D-ribofuranosyl) -2-thiouracil in amorphous form. According to thin-layer chromatographic comparison, this product was essentially identical to that according to the method of Brown et al., J. Chem. Üoc. 1- (2,3-ü-isopropylidene-β-D-ribofuranosyl) -2-thiouracil obtainable in 868 (19i? 7).

Ufi.ii of; Worthwhile procedures were repeated, with the early jjractories being caught with aoc sensitive component.

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Crystallization from ethyl acetate gave 1- (2,3-0-isopropylidene-β-D-ribofuranosyl) -2-thiouracil of melting point 194.5 Ms 19ö ° 0.

Jp "" 41 ° (° = ° »7276 in 95> ö ethanol) UV absorption:
75 * ethOH

7 * _{2ig 274 (fee 17}

iiJLct A .. /

CV '_ -1

Charak-tidst. iK-ATDSorptionshanden (cm ") NH / OH: 55oo, 32oo, 312o, 3o8o; G = O: 169ο; G = C / G = N: I64o, 162o, 149o; G = S / G-u / C-M / other: 1275, 1255, 122o, 155o, 111o, 1oSo, 1o7o; others: 89o, 835.

Analysis: Ber. for C ₁₂ H ₁₆ W ₂ U ₅ S: C: 47.98; H: 5.37; N: 9.33;

S: 1o, 68. Found: G: 47.92; H: 5.43; W: 9.43;

S: 1o _f 7o.

Part 13: 1-β-I) -Ki "bofuranosyl-2-thiouracil

i ^ a sample of the amorphous 1- (2,3-L'-isopropylidene-β-] J-ri'bofuranosyl) -2-thiouracil of fractions 3o "to 39 (see Part A) and 1 ml of 1N hydrochloric acid were refluxed for 4 minutes. The resulting aqueous acidic solution was "at reduced." Pressure evaporated to 'i' dry and the residue was in Water dissolved, then the "water" became under reduced pressure removed. After recrystallizing twice from water, 1-ß-D-K.i "bofuranosyl-2-thiouracil of melting point was obtained 2o6 to 2o9 ° C.

UV absorption: J ^ l * ^ä ° ^{Sa 2} 59; 27o.

'in et ever

Example 3

r-c of 1-i3 - U-uibot'uranosyl-2-thiothymine

Part A: 1 - ( Z, 3-ü-Isopro py "J iclen-ß- ü-ri bof uranosyl) -thy min

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A suspension formed with the exclusion of moisture 5.95 g (23.0 millimoles) 1-β-D-ribofuranosylthymine, 11.25 g anhydrous copper (II) sulfate, 23o ml of acetone and 0.23 ml concentrated sulfuric acid for 40 hours at 25 ° G. The bus pension was then covered by a layer of "Celite" (Diatomaceous earth filter aid) filtered, the filter cake was carefully washed with acetone. After union of the filtrate and the washing solution were 5.5 g of calcium hydroxide admitted. Ms mixture was 5 hours "at 25 C with exclusion Stirred by moisture, then filtered through a layer of Celite, the filter cake was again carefully with Acetone washed. The filtrate and acetone vash solution were then "concentrated to dryness under reduced pressure. The resulting white foam was azeotroped 2 with ethyl acetate under reduced pressure. The one left behind Solid was dissolved in hot methanol, then the methanol was removed under reduced pressure. The white, non-crystalline residue was then kept at 50 ° C. for 1 3/4 hours (Bath temperature) kept in a high vacuum, 7.0 g of 1- (2,3-ü-isopropylidene-β-D-ribofuranosyl) thymine were obtained.

Part B: 1- (5-0-i ^I osyl-2.3-0-isopropylidene-.beta.-D-i'ibofuranosyl) thymine.

6.56 g (22.0 millimoles) 1- (2,3-G-isopropylidene-β-D-ribofuranosyl) -thymine (see Part A) were dissolved in pyridine, then 4.61 g (24.2 millimoles) of the solution became freshly crystallized p-Toluenesulfonyl chloride added. The total amount Pyridine in the reaction mixture was 37 ml. The reaction vessel was tightly closed and left in the semi-darkness for 24 hours. After adding 3 ml of water to the mixture, became Left to stand again for 1 hour, then the mixture was poured into 140 ml of water with stirring. It was different a gummy substance, the supernatant aqueous liquid was acanted off. This liquid was 5 times with

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30 ml of chloroform was extracted, the chloroform extracts were combined and the gummy substance was in the extracts solved. The chloroform solution was successively 3 x with water, 3 x with 1 of the above aqueous potassium bisulfate solution, 2 with cold saturated sodium bicarbonate solution and 2 χ with ice water washed. Then the chloroform solution was treated with decolorizing charcoal for several hours, whereby von Has been shaken from time to time. The solution was then filtered through Celite. The carbon left on the filter was washed with chloroform, and the filtrate and washing solutions were dried over anhydrous sodium sulfate. Do that Removing the chloroform under reduced pressure, the foam-like residue was dissolved in hot 95% aqueous ethanol solved. The solvent was in a high vacuum at a bath temperature from 5o ° C away. The residue obtained was a further 2 χ with 95% aqueous ethanol in a high vacuum subjected to azeotroping and the resulting amorphous residue was left in a high vacuum for 2 1/2 hours held at 5oC. In this way, 8.42 g of 1- (5-0-tosyl-2,3-U-isopropylidene-β-D-ribofuranosyl) thymine were obtained.

Part C: 2,5'-Anhydro-1- (2,3-0-isopropylidene-β-D-ribofuranosyl) -thymine.

A solution of 1.0 g (2.21 millimoles) of 1-X5-0-tosyl-2,3-0-isopropylidene-ß-D-ribofuranosyl) thymine (see Part B), 12 ml of dimethylformamide and 1, above-mentioned sodium bicarbonate is heated with stirring in a nitrogen atmosphere at a bath temperature of 150 to IbO ⁰ C for 45 minutes. The reaction mixture is colored dark. It is then filtered and the filter is washed with methyl formamide, the filtrate and washing solutions are evaporated to dryness while the methyl formamide is evaporated off under reduced pressure. The residue is subjected to an azeotrope treatment 3 times with absolute ethanol at reduced pressure. The resulting solid will

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dissolved in warm 90% aqueous ethanol, then 2.5 g of silica gel are added. The slurry is concentrated to dryness under reduced pressure. The dry silica gel containing the desired compound is applied to a column of silica gel which is wetted with solvent A. The product is eluted with solvent A. After removal of the solvent from the jiluate under reduced pressure, a white substance is obtained from which, when crystallized from a mixture of absolute ethanol and technical hexane, 356 mg of 2,5'-anhydrous -1- (2,; JU-isopropylidene-ß-D-ribofuranosyl) -thymine with an undefined melting point can be obtained. UV absorption:
V ^ Ätun ^ _{46 (fes 12 4oo)}

Characteristic. lii absorption bands (cm ")

G = O / C = ii / O = G: 165 o, 1545; C-ü / C-w / others: 1295, 1285, 121o, 1155, 1145, 1o95, 1o65, 1o55; other 86o, 79o. Analysis: Calc. For Ο ^ ϋ ^ ϋ ^ Ο ^: C: 55.71; H: 5.75; N: 1o, oo Found: G: 55.89; H: 6.00; N: 1o, o9.

Part I): 1- (2,3-ü-Isopropylidene-ß-u-ribof \ iranosyl) -2-ü-methylthymine.

Line a warm solution of 26o mg (i, o millimoles) of 2,5'-anhydro-1- (2,3-iJ-isopropyliaen-ß-jj-ribofuranosyl) -thymine (see Part 0) and add 2 ml of methylformamide under tap water (about 25 ° C) cooled, then 20 ml of absolute methanol and 21 mg of sodium methylate are added. The reaction vessel solidifies closed and left to stand at 25 ° C for 17 hours. Then the solution is neutralized with the dropwise addition of glacial acetic acid and the liquid components are removed by evaporation at reduced pressure. The white i'eststoff thus obtained v / is carefully triturated with acetone, the acetone solution v / ird filtered and the filter is washed with acetone. Ji'iltrat and washing solutions (about 15 ml) v / ground with technical tiexan

209829/1156

diluted to cloudiness, whereupon crystallization occurs immediately. After the resulting mixture has stood at 25 ° C. for 16 hours, the crystals are collected on a filter, washed with a mixture of acetone and technical grade hexane and dried. 222.7 mg of 1- (2,3-0 ± sopropylidene β-D-ribofuranosyl) -2-O-methylthymine with a melting point of 164.5 to 160 ° C. are obtained.
UV absorption:

Ιΐ ^AtÜH

231 (üch); 252 m / u (6 = 9,000; II700)

mm i

Characteristic. IK absorption bands: (cm ~) OH: 3250; C = ü / ü = N / C = G: 1655, 16o5, 157o (black), 152o. U-u / G-iV'other: 129o, 127o, 124o, 121o, 1155, 1125, mo, 1o95, I060, 1o25, Ι0Ι0; others: 87o, 785.

Analysis: Ber. for ^ ₁₄ H ₂₀ N ₂ O ₆ : C: 53.84; H: 6.45; M: 8.97. Found: G: 53.82; H: 6.22; Li: 8.87.

'i'eil i->: 1- (2,3-U-isopropylidene-β-D-ribofuranosyl) -2-thiothymine.

A solution of 158 mg (0.5o5 millimoles) of 1- (2,3-0-isopropylidene-β-D-ribofuranosyl) -2-0-methylthymine (see Part D), 5 ml of dimethylformamide and 0.4 ml of triethylamine is treated with hydrogen sulfide treated by slowly passing a stream of the gas through the reaction mixture. The treatment lasts 18 hours at 25 ° C. Excess hydrogen sulfide is then removed by purging with nitrogen gas, and the like Hydrogen sulfide-free reaction mixture is through a ! b'ilter poured into a flask and the volatile components are evaporated under reduced pressure. The residue is 3 times with absolute ethanol at reduced pressure Subjected to azeotroping treatment. The resulting solid is slurried with silica gel and the slurry becomes applied to a column with gel gel, which with solvent A is wetted. The column is developed with solvent A, collecting 1 ml fractions. the

209829 / 11B6

Actions 41 "to 48 are combined and the solvent A is evaporated off under reduced pressure. The resulting solid is recrystallized from a mixture of acetone and technical grade hexane, 51 »4 mg of 1- (2,3-0-isopropylidene-β-D-ribofuranosyl) -2-thiothymine having a melting point of 215 to 215 ° C. receives.
UV absorption:

²⁷⁹ ' ^{5 m} / ^u <£ = 16150, 17555)

uharacterist. Ik absorption bands (cm) ÜH / Kh: 353o, 333o, 322o (Sch); C = U / C = C: 167o, 16o5, 149o; C-O / G-N / others: 1295, 1275, 127o, 123o, 122o, 1i6o, 11o5, 1o9o; others: 85o.

Analysis: Ber. for C ₁₅ H ₁₈ N ₂ O ₅ S: C, 49.67; H: 5.77; N: 8.91; S: 1c0: Found: C: 49.3o; H: 6.25; H: 8.55; S: 1o, o9

Part Έ: 1-ß-I) -rihofuranosyl-2-thiothymine.

Solution of 300 mg (0.955 millimoles) of 1- (2,3-0-isopropylidene-β-i) -ribofuranosyl) -2-thiothymine (For preparation see 'feil L ·) and 4.5 ml 90 / o aqueous trifluoroacetic acid in one tightly closed flask is heated for 10 minutes at 25 ° G let react. The volatile components are then removed under reduced pressure and the residue is washed 3 times with subjected to an azeotropic treatment of absolute ethanol at reduced pressure. The solid residue thus obtained is under Using methanol as washing liquid transferred into a clean flask, then das.Methanol at reduced Pressure removed. The pest is rubbed with absolute ethanol and the ethanol is evaporated off under reduced pressure, thus obtaining a crystalline solid which, at 55.degree (Bath temperature) is kept under high vacuum for several hours. The evacuated crystals melt at 198 to 2o1 G. .beim recrystallization from a mixture of. absolute ethanol

20982 9/1156

With technical Mexan, an analytically pure sample with a melting point of 2o6 to 2o8.5 ö is obtained.
/ 7x_7 UV absorption:

_7 -. + 1 (c = o, 4772 in! Dimethylformamide)

^m / ^u (^ = 20000, 2οδ5ο, 19900)

Characteristic. Ift absorption bands (cm ") Nri / OHs 346ο, 342o, 335o, 326o; C = u / O = Ü: 1685 (öch), 1665, 149o; Gi ^ i / O = S / Gu / others: 1265, 1235 , 1165, 113o, 11oo, 1o8o 1o55, 1o4o.
Analysis: Ber. for C ₁₀ H ₁₄ N ₂ U ₅ Si C: 43.78; H: 5.14; M: 10.22;

S; 11.68. Found: C: 44.18; H: 5.9o; N: 10.27;

S: 11.42.

Example 4

Production of 1- (2-! Üeoxy-ß ~ I) -er.ythropentofuranosyl) -2- »thio ° thymine and its 3t5 - m - C

ΐβϋ A: 1- (2-Deoxy-ß-lt-erythropentofuranosyl) -2-Ü-methylthymine _e

To a solution of 41 ο ml of absolute methanol and 1.12 g (5, ο Mllimol) 2,5'-anhydro-1- (2-deoxy-β ~ I) -eryh ^ ropentofuranosyl) -thyinin (prepared according to the method von Michelson and l'odd, J. Chem. boc, 816 (1953)) 4 ml of methanol, which had been saturated with anhydrous ammonia at from above to -5.degree. C., are added. The flask is tightly closed, left to stand at 25 ° Θ and the course of the reaction is followed by I) thin-layer chromatography or the mixture of "solvent C" (mixture of ethyl acetate, technical hexane and methanol in a ratio of 5: 2: 3), jtfach 12o hours after the reaction only an amount of 3o ^c; _s has reached £ wei the more

209829/1156

7 ml of saturated methanolic ammonia were added. After a further two weeks of reaction time, after the reaction appeared to be complete, methanol and residual ammonia are removed with reduced bridging. The residue is subjected to an azeotrope treatment with absolute ethanol at reduced pressure for 2 hours and the white crystalline substance thus obtained is kept under high vacuum for 2 hours . 1oo mg of crystals are recrystallized from a mixture of absolute methanol and absolute ether, to yield the 1- (2-deoxy-.beta.-D-erytnropentofuranosyl) -2-u-methylthymine of melting point 14 ", 5-151 ⁰ O .
UV absorption:

^ maxf ^ ^{255 (üch)} 5 ^{252 m} / ^{u (6 = 88o} ° ' ^{1o5i? o)}

__ -ι characteristic. IR absorption bands (cm) ÜH / = CH: 342o, 33So, 3o6o, 3o8o; G = ü / C = C: I65o, I6io (Sch), 16oo, 157o, 1515, 1485; C-O / C-tt / others: 129o, 119o, 111o, 1o75, 1o3o, 1o1o, 1ooo; others: 845, 785.

Analysis: Ber. for CjJ ^ gN ₂ ⁰ V ^{C: 51} > ⁵⁶ ' ^{H: 6} » ²⁹ » ^W: 1 ° »93. Found: C: 51.71; H: 6.15; N: 1o, 88

Part B: 1- (2-jjeoxy-β-U-erythropentofuranosyl) -2-thiothymine.

With the exclusion of moisture, a suspension of 19o mg (o.742 i-iillimol) 1- (2-ueoxy-β-JJ-erythropentofuranosyl) -2-u-methylthymine (see section A), flushed with a stream of dry nitrogen gas in 6 ml of methylformamide, ran 0.37 ml of triethylamine were added, and it was still Flushed with dry nitrogen for 10 minutes. Then the nitrogen purge was stopped and it became a slow one Hydrogen sulfide stream initiated for 26 hours. After this treatment there was still a small amount of undissolved material Starting compound before, however, the reaction mixture was transferred into a clean customer flask, with dimethyl

209829/1156

formamide was rinsed. The dimethylformamide was then "distilled off under reduced pressure and the residue was subjected to an azeotrope treatment 3 times with anhydrous benzene" at reduced pressure. The semi-solid residue thus obtained was carefully triturated with acetone and the acetone-insoluble components were filtered off. The acetone was evaporated from the filtrate and 147.5 g of a semicrystalline substance were obtained. This was chromatographed on 18.5 g of silica gel using solvent G for development, collecting 1.5 ml fractions. Fractions 19 to 25 were combined and evaporated. A practically white crystalline residue was obtained, which was taken up in methanol. After pooling the fractions in hethanol and evaporation of the methanol under reduced pressure gave 35.2 mg of 1- (2-deoxy-.beta.-D-erythropentofuranosyl) -2-thiothymine of melting point 173-175 ° C _O UV absorption:

²² ° 5 ^{278 m} / ^{u (e} = ¹⁵²⁵ ° 'Ί7 ° οο) ) ° tl ^{n Na0H} 241; 265 (black Sch) m.-u ( G = 25 8oo, 14 95o)

'* - QlcL X. /

Characteristic of IR absorption bands (cm) IMH / üH / = CH: 314o, 3o9o, 3o6o; C = ü / G = C / ü = N: 1695, I68o, 1635 (Schw.Sch), 15oo; G = S / G-N / G-ü / other: 131o, 128o, 123o, 1185,

1145, 11o5, 1o7o, 1o6o, 1o1o

Analysis: iier. for G ₁₀ H ₁₄ N ₂ O ₄

Found:

The above preparation was repeated on a 2.5 fold scale A final recrystallization from a mixture of absolute ethanol and technical grade hexane gave the compound from melting point 175 to 176 ° C

G: 46.5ο; H: 5 , 96; N: 1 ο .85; S: 12.42. G: 46.6ο; H: 5 , 8ο; N: 1 0 , 07? S: 12.48.

209829/1156

i ~ ^ 7p ⁵ + 44 ° (0 = o, 4322 in o, 1-n NaOH) UY absorption:

^ mIx. ^221; 277 m / rev (£ = 151oo, 1685o). IR absorption (cm ""): as above
i-mass spectrum: M ⁺ 258; B +1, 142
Analysis: Found: G: 46.39; H: 5.24; N: 11.12; S; 12.12.

li due to the rapid cleavage with üscherichia coli phosphorylase the u-anomeric configuration was confirmed.

Part ü: 1- (3,5-iitü-acetyl-2-deoxy-ß-D-erythropentofuranosyl) -2-thiothymine.

i. a suspension of 516 mg (2.0 millimoles) of finely divided 1- (2-üeoxy-ß-I) -erythropentofuranosyl) -2-thiothymine, 4.0 ml of acet- ■ -: .ihydride and 2 drops of pyridine was added under Exclusion of moisture on a bath at a temperature of 76 to 80 s .; wa heated for 72 hours. The reaction vessel was then allowed to cool slowly, after which it was poured into a mixture of crushed lis and water. The reaction vessel was rinsed with a little pyridine, and the filling solution was combined with the main amount. l) The crystallization took place spontaneously by rubbing the walls of the reaction vessel. The reaction product, which crystallized in the mixture of Lis and water, was kept undisturbed for 11/2 hours, then it was cooled to 3 ° C. for 5 hours. The crystals were then filtered off and the filter cake was first mixed with the mother liquor and then washed with ice-cold water. When the crystals were dried in air, 565 mg of 1- (3 »5-di-O-acetyl-2-deoxy-β-i) -erythropentofuranosyl)» 2- "thiothymine with a melting point of 155 were obtained to 160 ° G. About 545 mg of the compound were recrystallized from a mixture of absolute ethanol and technical grade hexane, after decolorization with charcoal, whereby 49o mg of the analytically pure compound with a melting point of 152-164.5 G

209829/1156

were received «
_ " ■ - ■ -; ο η Z ⁰ L / '-λ * -'i4 \ ö = o.7192 in acetone /

UV absorption:

j ⁹⁵ ' ^{J Atüxl} 221; 277 m, u (6 = 1535o, 1ö4oo)

■ - max. /

üharakteris'c »Ixt absorption bands (cm") Wn / = Ch: 516ο, 3ö8o; O = Oj 17415, 173o; O = u / O = N / C = ü: 1675, G-ü / C-ü / aiidere : 13oo, 129o, 12öo, 126o, 1245, 1225, 12oo _? 1i6o _t 11o5, 1o95, 1o25, 96o.

Analysis: Ber. rlir U ₁₄ H ₁₈ N ₂ U ₆ L: o: 49.1o; rl: 5.3o; N: 8.19?

ü: 9.37.

Found: G: 49.2o; H: 5.89; M: 8.19;

ö: 9.13,

Example ρ

Manufacture of 1- (2-Leox; / - ß-D ~ erythropentofuranosyl) -2-tiiiour acil and aess en 315-i ^ iacetat .

Rope A: 1- (5-0-Tosyi-2-deoxy-ß-D-erythropentofuranosyl) -uracil <

With exclusion of moisture, a solution of 1.18 g (5.2 millimoles) of 1- (2-i> eoxy-β-1) -ribofuranosyl) uracil in 6 ml of pyridine is cooled to 50 ° with stirring. At this temperature 1.20 g (0.63 millimoles) of p-toluenesulfonyl chloride, dissolved in 6 ml of pyridine, are added. The reaction vessel is tightly closed and then left to stand at 3 to 5 ° G for 3 days. Then excess p-'l'oluenesulfonyl chloride is decomposed by adding crushed .ti3. As soon as the .bis has melted, the reaction mixture is poured into a separating funnel, which contains 80% water. The diluted aqueous solution is then mixed 3 times with each 25 ml of chloroform extracted. The combined chloroform extracts are mixed 1 χ with pure water

O 8 8 2 9/1 1 S S

washed and then dried over anhydrous matrix sulfate. The chloroform is evaporated off under reduced pressure and the residue is subjected 3 times to an azeotropic treatment with 95% aqueous ethanol, a crystalline white solid being obtained. Recrystallization from a mixture of acetone and technical grade hexane gives 1, og 1- (5 - ^ - ^r i'osyl-2-deoxy-ß-D-erythropentofuranosyl) uracil.Recrystallization of a 150 mg sample from the mixture of acetone and technical hexane yields the analytically pure sample with a melting point of 158 to 158 , 5 ⁰ C. UV absorption:

^{223; 261 ra} / ^u ( ^{t = 1} 535o, 1o15o)

-1 ) Characteristic. III * absorption bands (cm ^; NH / OH / = CH: 338o, 316o, 31 oo, 3o6o; C = O: 172ο, 167ο; C = C: 1625,

1575; -SOjü / C-O / O-N / others: 1365, 127o, 119o, 118o, 1175, 1o9o,

1o65, 92o: p-CH / others: 825, 815, 81 ο.

Analysis: Ber. for C ₁₆ H ₁₀ N ₂ O ₇ S: 0: 5o, 3o; H: 4.75; N: 7.33;

S: 8.39.

Found: C: 50.63; H: 4.98; IM: 7.08;

S: 8.12.

Part B: 2,5'-anhydro-1- (2-deoxy-β-D-erythropentofuranosyl) uracil.

In a dry nitrogen atmosphere, with exclusion of moisture, a reaction mixture of 4.43 g (11.6 millimoles) 1- (5_O- ^l isyl-2-deoxy-ß-1) -erythropentofuranosyl) -uracil (see! • Part A) , 150 ml of dimethylformamide and 5, og sodium bicarbonate heated to 145 ° C for 50 minutes. The mixture is then cooled to 25 ° C., filtered and the i'lltrat saved. The dimethylformamide is removed under reduced pressure and the residue is left. is subjected to an azeotrope treatment 2 χ with absolute ethanol at reduced pressure. The residue is dissolved in 95% aqueous hot ethanol, the solution becomes

209829/1156

10 g silica gel added. By evaporating the ethanol under reduced pressure, a dry mixture of reaction product and silica gel is obtained, which is applied to a column with 275 g of silica gel which had previously been wetted with solvent G. The chromatogram is developed with solvent C. After removing the solvent from the oil, 1.4o g of crude 2,5'-anhydro-i- (deoxy-β-D-erythropentofuranosyl) uracil are obtained. When a 100 mg sample is recrystallized from aqueous ethanol, the analytically pure sample with a melting point of 182 to 184 ° C. is obtained.
UV absorption:

/ LmIx. ^{237; 255 (bcil) m} / ^{u (e = 12} ° ⁰⁰ ' ^{7 8oo)}

Characteristic. IR absorption bands (cm) OH: 338o, 329o; G = O / C = C / C = N: 1645, 162ο (Sch), 1615, 151o, G-ü / C-N / other: 126o, 11 To, 1115, 1o9o, 1o8o; Aroma. GH / others: 86o, 835.

Analysis: Ber. for C ₉ H ₁₀ N ₂ O ^: C: 51.43; H: 4.80; N: 13.33. Found: G: 51.19; H: 4.45; N: 12.87.

l ^l eilC: 1- (2-Deoxy-ß-D-erythropentofuranosyl) -2-U-methyluracil.

A reaction solution of 26o, 9 mg (1.24 millimoles) of 2,5'-anhydro-1- (2-deoxy-ß-D-erythropentofuranosyl) uracil (part 15), 3 ml of dimethylformamide, 165 ml of absolute methanol and 3 35 ml of triethylamine are tightly closed and left to stand at 25 G for 3 l'age. Solvent and volatile components are then removed by evaporation under reduced pressure. The residue is subjected to 3 χ an azeotrope treatment with absolute ethanol, the ethanol being removed under reduced pressure. The residue obtained in this way is dissolved in 95 ^{l of} aqueous ethanol, then 1.5 g of silica gel are added to the solution. After removing the aqueous ethanol under reduced pressure, the mixture of bilikagel and reac-

209829/1156

tion product on a column with 35 g of silica gel, which had been wetted with solvent 0, applied. The column is developed with solvent C, then the solvent is removed from the fraction containing the desired compound under reduced pressure. The residue is dissolved in ethanol. After the insoluble fractions are filtered off, the methanol is removed from the piltrate under reduced pressure, 1 ~ (2-deoxy-β-D-erythropentofuranosyl) -2-0-methyluracil having a melting point of 134.5 to 136 ⁰ G being obtained . uY absorption:

^{22Ö; 2} ^ om / u (έ = 113οο, 1015ο) -

Oi characterist Iit absorption bands (cm)

I.ii / üii: 34oo, 500O (broad), 272o (Sch); C = ü / C = H / C = C: I640,

161 o, 1500; C-ü / C-iM / other: 129o, 1115, 1100, 1o8o _s 1o3o, 1c »15; others: 835

Analysis: Ber. for O ₁₀ H ₁₄ H ₂ O ₅ : G: 49.58; H: 5.83 ί Hs 11.57ο

Found: G: 49.31; ti: 6 ₉ o2? Ms 11.42.

ϊβχΐ J): 1- (2-deoxy-ß-D-erythropen.tofuranosyl) -2-thiouracilo

i-Ine solution of 1- (2-deoxy-β-D »erythropentofuranosyl) -2-u» Kethyluracil (for preparation see Part G) in 50 ml of dimethylformamide, which contains 3.3 ml of triethylamine, is treated with sulfur water = CTOf ± "treated by lr 17 hours a flow of the gas:? ig slowly passed into the solution Here, moisture is kept away, the l'emperature is 25 ° 0 ₄ the Keaktionsge- Y'iLsch v / ith then with dry nitrogen.. rinsed _{9 in} order to drive out any excess sulphurous water. Then the mixture is transferred to a clean round-bottomed flask, where it is rinsed with a large amount of dimethylformamide. The dimethylformamide is then reduced at reduced The pressure is removed and the residue is subjected to an azeotrope treatment at reduced pressure with absolute ethanol

2 O 9 8 2 S / 1 1 5 6

dissolved in hot absolute ethanol, then 5, og oil gel are added to the solution. After removing the ethanol by evaporation under reduced pressure, the dry mixture is applied to a column with 125 g of oilikagel, which has been prepared with a slurry of oilikagel in solvent A. The column is developed with solvent A, collecting fractions of 10 ml each. The fraction 41 to 59 are combined and freed from the solvent under reduced pressure. A white solid is obtained which is crystallized from a mixture of absolute ethanol and technical iiexane, 1- (2-deoxy-β-jjerythropentofuranosyl) -2-thiouracil having an undefined melting point, which is apparently at 133 to 134 ° C , receives. £ "^ ίψ + 68 ° (C = 0.2524 in 0.1 n-NaüIi) UV absorption:

Ügi? 217; 274 m / u (b = 15ooo; 1295o)

λ max? ^{WaÜii 2} ^ ^{269 m} / ^u <t- 19 6oo; 12 6 ₅ o)

Max? ² ^ ^{269 m} /

mm i

Characteristic. lü absorption bands (cm ") WH / OH / = CH: 339o, 318o, 3obo; C = O / C = C / "C-N": 1675, 1655, 14Ö5, "C = S" / ΰ-ü / C-l · ί / others: 128o, 1195, 1165, 114o, 1115, 1o9o, 1o4o.

Part 1 · ;: 1- (3,5-JJi-0-acetyl-2-deoxy-β-D-erythropentofuranosyl) -2-thiouracil.

i a suspension of 488 mg (2.0 millimoles) of finely divided 1- (2-jjeoxy-ß-D-erythropentofuranosyl) -2-thiouracil (see 'feil D) in 4 ml of acetic anhydride, which contains 2 drops of tyridine, is placed on a J3ad at a temperature of 75 to 76 O for 71 hours. warmed up for a long time, excluding moisture, etc. Reaction mixture is then cooled and converted into a mixture of his and V / ater poured. Lying and rubbing the walls of the reaction tank, crystallization occurs spontaneously. That

20982 9/1156

crystallizing mixture is left to stand for a few hours, occasionally rubbing. After cooling down to 3 ° C the crystals are collected on an irilter, the filter cake is washed with the mother liquor and then with white water and dried. 43o + mg of 1- (3,5-M-0-acetyl-2-deoxy-β-D-erythropentofuranosyl) -2-thiouracil are eaten. xi by recrystallization from a mixture of absolute ethanol and technical hexane under decolorization with charcoal are 4oo mg an analytically pure sample with a melting point of 13o to 131 ° C obtain.

°° ( ^G = o, 552o in acetone)

ÜY absorption:

1 95; _: EtOii _{274 m}

Ghai characteristic. liv absorption bands (cm ~) im: 321o, 316o, G: 0: 1750, 173o, 17o5; O = G / O = N: 162o, 1485; C = S / C-ü / GN / other: 131o, 128o, 125o, 1195, 114o, 1115, 1o8o, 1o55, 1o25; Aroma. Cu / other: 835.
hate spectrum:

M ⁺ 328; B +1 128; Jeoxy sugar " ¹ " 2o1.

Analysis: Ber. for G ₁₅ H ₁₆ N ₂ O ₆ S: C: 47.55; Ii: 4.91; K: 8.53; S: 9.77 Found: G: 47.1o; H: 4.86; H: 8.62; S: 9.58

Example 6 Preparation of 1- (2-jeoxy-ß-Li-threopentofuranosyl) -2-thiothymine

Part A: 1- (5-0-'i ^l osyl-2-deoxy-β-D-three open of uranosyl) -thymine.

With the exclusion of i'Ouchtigkeit is at a temperature of etv / a O ⁰ G, a solution of 1.18 g (6.2 Mllimol) of p-toluenesulfonyl chloride in pyridine to α ml uUhren to 1,2o g (4.96 millimoles; practically completely dissolved 1- (2-Deoxy-ß-ii-threopentofura-

BATH ORIGINAL 209829/1156

- 3ο -

nosyl) thymine added. The reaction mixture is then tightly closed and cooled to 3 to 5 ° C. overnight. The next I ¹ ag a small amount gestoi3enes i '^ is added, then another 2 hours is still cooled at 3 to 5 C. The reaction mixture is then poured into 200 ml of ice water, the flask is rinsed with 75 ml of water and the rinsing solution is added to the main mixture. The aqueous solution is then poured into a separating funnel and extracted 3 with 25 ml of chloroform. The chloroform extracts are combined, washed 3 times with saturated aqueous sodium bicarbonate solution and 1 with water and dried over anhydrous sodium sodium sulfate. After removing the chloroform at reduced. Pressure and 5 times azeotroping of the residue with absolute ethanol at reduced pressure gives a practically pure compound, as determined by thin-layer chromatography using solvent A. When the solids crystallize from a mixture of acetone and technical grade hexane, 1.51 g of 1- (5-P-'i'osyl-2-deoxy-iu-D-threopentoi \ xranosyl) -thymine with a melting point of 163 to 164 are obtained ° G received.
UV absorption:

224; 266, 272 (Sch) m / u (b = 1d7oo, 975o, 9ooo) -

- 1 characteristic. IK absorption bands (cm ~)

ün / = Cri: 342o, 31ao, 3o5o; O = ü / ö = C / other: 1715, 1695, 16? O, G = U: 16oo; -bO ₂ -ü / C ^l -O / others: 137o, 1355, 1275, 1195, I'loo; lin identified: 9β5, 965 _S ; Aroma. Üii / others: V> bo, 775. · Analysis: At ·, for G ₁₇ H ₂₀ M ₂ (J ₇ S: G: 51.56; H: 5, o9; H: 7, o7;

S: b, 1o.

Found: G: 52, oil; Ii: 5.46; 1 \ 1: 7, o2;

S: ti, 36.

ulle B: 2,5'-anhydro-1- (2-deoxy-β-D-threopGntof ui'anosyl) -tliymine. In a nitrogen atmosphere, a U-emiach becomes au »üoo mg Ί - ν. ! ν — Ο—

BATH ORIGINAL

209829/1 156

Tosyl-2-deoxy-ß-D-threopentofuranosyl) thymine (see Part A), 3o ml of dimethylformamide and 2 g of sodium bicarbonate under heat on a xiad from 130 to 150 ° C for 75 minutes »Then the reaction mixture is cooled and filtered . The filter is washed with dimethylformamide, the filtrate and ash solutions are combined, then. After evaporation of the dimethylformamide under reduced pressure, the residue obtained in this way is dissolved in aqueous ethanol, and 2 g of bilikägel are then added to the solution. The ethanol is evaporated off under reduced pressure and the mixture obtained in this way is applied to a column with bilikägel which had been wetted with solvent G. After developing the chromatogram with solvent C and collecting the fluid, the solvent is evaporated off under reduced pressure. The residue is subjected to an azeotrope treatment with ethanol under reduced pressure. The residue obtained is crystallized from absolute ethanol, and xvaä under reduced pressure. Dried 40 ° G, 179 mg of 2,5'-anhydro-1- (2-deoxy-β-D-threopentofuranosyl) .- thymine having a melting point of 228 to 23 ° G are obtained.
UV absorption:

rlmax. ^At0H ^ o (se); 246 nyu (G = 815o, 82oo)

Characteristic, lii absorption bands (cm) üü: 3530, 33üo, 32bo; G = ü / ü = N / G = G: 1665, I630, 1535, 1485; G-O / C-W / others: 1275, 114o, I080, 1o5o, 975, 9oo, 88o, 8I0, 79o.

Analysis: Ber. for C ₁₀ H ₁₂ W ₂ O ₄ : C: 53.57; ti: 5.39; N: 12.5o. Found: C: 53.49; ü: 5.81; N: 11.80.

Part C: 1- (2-Deoxy-β-jj-threopentofuranosyl) -2-0-methylthymine.

To a lukewarm solution of 224.0 g (1.0 millimoles) of 2,5'-anhydro-1- (2-deoxy-ß-D-threopentofuranosyl) thymine (see part ιϊ) in a solvent mixture of 2 ml of dimethylformamide

i, * _Ä . 2 0 9 8 2 9/1156 BAD ORIGINAL

and 2 ml of absolute ethanol, 18 mg (0.323 millimoles) of sodium methylate are added, then the reaction vessel is tightly closed and left to stand at 25 ° G. After several days, a further 150 mg (2.78 millimoles) of sodium methylate are added, and after a further 6 days the reaction mixture is neutralized by the dropwise addition of glacial acetic acid. The solvents dimethylformamide and methanol are removed under reduced pressure and the residue is subjected to 2 azeotrope treatments with absolute ethanol under reduced pressure. The residue obtained in this way is dissolved in hot 95% aqueous ethanol and 2.5 g of bilica gel are added to the solution. The aqueous ethanol is evaporated off under reduced pressure and the dry residue mixture is applied to a column with 25 g of silica gel, which had previously been wetted with 4 ml of solvent C, - the column is developed with solvent C. Upon removal of the solvent from the üluat under reduced pressure to 116.7 mg of 1 - (2-deoxy-.beta.-D-tiireopentof uranosyl) -2-Ü-methyl-thymine obtained 159-161 ⁰ C of melting point.
UV absorption:

λ mS

Gharalcterist. Li absorption bands (cm ") Uli: 34oo, 31oo, 276o (Sch), 266o (Sch); U = u / G = M / O = C: 1b6o, 1615, 1525, 1485; C-u / G-N / others: 1295, 1255, 1225, 119o, 114o, 111o, 1o75, 1o6o, 1o1o, 99o; others: 87o.

Part D: 1- (2-Deoxy-J3-D-threopentofuranosyl) -2-thiothymine.

L'a solution of 106 mg (0.415 millimoles) 1- (2-Deoxy-13-D-threopentofuranosyl) -2-U-methylthymine (see Part G), 5 ml of dimethylformamide and 12 drops of triethylamine is 22 1/2 hours treated for a long time with a stream of hydrogen sulfide gas bubbling slowly through the solution. Then be Dimethylformamide and volatile components of the reaction

209829/1156

mixed evaporated at reduced pressure. The residue is subjected to an azeotrope treatment with absolute ethanol at reduced pressure. After thin-layer chromatography had shown that a large amount of the starting material was still present, this residue was dissolved in warm dimethylformamide, then 7 drops of triethylamine were added and treatment with hydrogen sulfide gas was 2 I ¹ age continued for. the reaction mixture was then concentrated at reduced pressure and the! residue was subjected to absolute ethanol at reduced pressure of a azeotropation. the iiückstand thereby obtained was dissolved in 95 / * aqueous ethanol, 1 g of silica gel was added to the solution. After evaporation of the aqueous ethanol under reduced pressure, the dry mixture was applied to a column containing 25 g of silica gel which had been wetted with solvent G. The column was developed with solvent G, whereby - Fractions of 3 ml each are collected . Fractions 14-17 were combined and the solvent was evaporated under reduced pressure. The residue was crystallized from a mixture of acetone and ethyl acetate, the 1- (2-deoxy-ß-D-threoperrtofuranosyl) -2-thiothymine having a melting point of 168.5 to 171 ° G (re-solidification at 175 ° C) received.
UV absorption:

) 95> Ätüii _{2 2 28Ö} ( _black> ^ ₀ J ₁ ) (£ _e -J ₄₀ Uj ₀ (V 'max · /

14900, 1315ο).

Characteristic. IR absorption bands (cm "") WH / Oh: 3410, 325o; O = u / O = G / O = W: 169o (black), 1fai? O, 151o, 1485; G-w / G = S / G-0 / other: 13oo, 1265, 1265, 113o, 11o5, 1o6o, 1o3o.

BAD 209829/1158

Example 7

Preparation of "1- (3-Deoxy-β-D-threopentofuranosyl) -2-thiouracil 'i'eil A: 1- (5-Ü-l ¹ osyl-5-deoxy-β-ii-threopentoi'uranosyl ) -uracil

With the exclusion of moisture, a solution of 228 mg (1.0 i'iillimol) 1- (3-deoxy-β-D-threopentofuranosyl) uracil (prepared by the method of iiorwitz et al., J. Urg. Uhem. 51, p. 2o5-211 (1966)) in 2.5 ml of l-yridine is cooled to ⁰ G, then a solution of -236 mg (1.24 i-iillimol) p-'x'oluenesulfonyl chloride and 2, 5 ml of pyridine were added dropwise, the temperature of the reaction mixture being kept at 0 °. The reaction vessel is then tightly closed and left to stand for about 16 hours at 3 to 5 ° C, then a small amount of crushed oil is added and it is cooled for about 1 to 2 hours. The mixture is then diluted with 50 ml of ice-cold water and the diluted aqueous solution is extracted 3 times with 30 ml of chloroform each time. The Oiilox ^v oformextrakte are combined and dried over anhydrous sodium sulfate. After evaporation of the chloroform under reduced pressure, the residue is dissolved in hot absolute ethanol. Uncolouring carbon is added to the ethanolic solution, this is filtered off and the filter is washed with hot absolute ethanol. i ⁽ⁱ iltrat and washing solutions are freed at vex-niindertem pressure from the ethanol and the üückstand is subjected with absolute ethanol at a reduced pressure azeotropation. thus obtained 182 U mg-1- (5-0-tosyl-3-deoxy- ß-D-threopentofuranosyl) uracil in i'Orm of an amorphous solid.
UV absorption:

^ ÄtOH ₂₃₅ ^ _{262j 272} ( _{Bchw # üch} ) ((ύ ₌ 155ΟΟ, lo '^ oo, 79oo)

Max. .

Characteristic. lit absorption bands (cm ")

BAD ORlGiMAL

20 98 29/1 1 pp

Nti / üH: 334o (wide); C = O / G = C / G = lSii 17o5 (ScIi) ₅ 1685, 1625 (Sch),

1595 (black); -Sü ^ ü / C-ü / CN / others: 156o, 128o ₅ 11So, 1175, 111o, 1o95, 1o85, 955.

Analysis: Ber. for. O ₁₆ H ₁₈ M ₂ U ₇ S: C; 5o, 3o; H: 4.74? N: 7.38;

-S: 8.38.

Found: C; 5o.89; H: 5.1o; Ns 6.665

S: 8, o7.

Part B: 2,2'-ArL "nydro-1- (3-d.eoxy-13-D-threopentofur.anosyl) -uracil.

In a reaction mixture of 1.54 g (4.03 millimoles) 1- (5-0-tosyl-3-deoxy-ß-D-threopentofuranosyl) uracil (for preparation see section A), 1.5 g sodium bicarbonate and 50 ml of diethylformamide is heated with stirring in the presence of nitrogen gas for 65 minutes at a bath temperature of 15 ° C. After cooling to about 25 ° C., the mixture is filtered and the filter is removed Washed with dimethylformamide, the dimethylformamide is evaporated off practically completely under reduced pressure and the residue is subjected to an azeotrope treatment 3 times with absolute ethanol at reduced pressure 5 g of silica gel are added to the solution, the aqueous ethanol is removed from the slurry under reduced pressure, and the dry mixture is applied to a chromatography column with 100 g of silica gel which has been wetted with solvent C. The column is applied with solvent O develop ckelt and the JJluat is caught. After the solvent has been removed under reduced pressure, the residue is crystallized from a mixture of absolute ethanol and technical grade hexane, then the crystals are dried at 40 G and reduced pressure, giving 160.3 mg of 2,2'-anhydro-1 - (3-deoxy-.beta.-D-threo-pentofuranosyl) uracil of melting point 214-216 ⁰ C is obtained.
UV absorption:

2 0 9 8 2 9/1156 ^{ßAD Of} «GINAL

^ -mix 22i ^{>> 255; 26} ° ^ ^Scll )> ²⁶⁹ ( ^Sch ) ^m / ^u ( ^{έ = 8} 35o, 6.1 oo,

625o, 24oo)

Characteristic, lit absorption bands (cm "):

OH: 332o; C = o / C = N / C = C: 1475; C-O / C-N / others: 124ο, 1ο95, 1ο4ο; = Cix / others: 83ο

Analysis: Ber. for CqH ₁₀ N ₂ O .: C, 51.42; E: 4.8ο, · N: 13.33; Found: O: 51.49; H: 5, ο4; N: 13.65;

Part 0: 1- (3-eoxy-β- "jj-threopentofuranosyl) -2-thiouracil.

Line solution of 42 mg (0.2 millimoles) of 2., 2'-Anliydro-1- (3-deoxyß-D-threopentofuranosyl) uracil (see Part B), 2 ml of dimethylformamide and 0.1 ml of triethylamine is treated with hydrogen sulfide treated by bubbling a stream of gas slowly through the solution for 114 hours. Additional triethylamine is added from time to time. Excess hydrogen sulfide is then removed by flushing the reaction solution with nitrogen, and the dimethylformamide is then evaporated off under reduced pressure. The residue is subjected to an azeotrope treatment with absolute ethanol at reduced pressure for 2 * The residue obtained is dissolved in 5 ml of 5% aqueous ethanol and 1 g of silica gel is added to the solution. The resulting slurry is freed from the aqueous ethanol under reduced pressure. The dry mixture is applied to a chromatography column with 18 g of silica gel which has been wetted with solvent A. The column is developed with solvent A and the fraction containing the desired compound is freed from the solvent under reduced pressure. The residue is made from a mixture of absolute ethanol and. technical hexane crystallized. This gives 26.6 mg of 1 - (^ - jjeoxy-ia-D-threopentofuranosyl) -2-thiouracil with a melting point of 6ΐ-163 ° ΰ. UV absorption:

95 '/ · »^ to OH _22o . _{27! 3} . ₂₈₅ ( _{schw # bcJ1} ) _{m / U} (^ ₌ Ib75o, 142oo, 138üq

2098 29/1156 BAD OR.SINAL

- ■ 37 - '

Characteristic. Ιϊί absorption bands (cm ") üh / Wii: 359ο ·, 32oo; G = O: 167o; δ = fl / C = C: 1495; ü = S / C-G / others:

1285, 115o, 1115, 11oo; others: 82o.

Analysis: Ber. for G ₉ H ₁₂ I ₂ O ₄ S: G: 44.25; H: 4.95; N: 11.47; '- J

S: 13.13. Found: G: 44.3a; H: 4.79; N: 12.28;

S: 12.67.

Example 8

If the procedure of Example 4, Part G is repeated, however using the starting materials 1-ß-i) -Ki'bofuranosyl-2-thio-b-aiiauracil, 1-ß-jj-iiiboiuranosyl-2-thiouracil,

i-ß-ij-kibofiiranosyl-2-thiothymine,

1- (2-ueoxy-β-ij-threopeiitofuranosyl) -2-thiothymine or 1- (3-.ueoxy-β-jj-threopentofuranosyl) -2-thiouracil, so obtained man the connections

1- (2,3,5-'i ¹ ri-0-acetyl-iJ-jJ-ribofuranosyl) -2-thio-u-azauracil with the optional direction

J ² J ^j0 -4-4- ° (G = 1, o668 in

I) V absorption:
1 ethanol _{21? 2? 1} ^ ^ ₅ (black Öcla) mu (G = 14I00; 17ö5o;

2o5o)

Gharakberist. iü absorption bands (cm ") Oh: 34oo (broad); WIi / oi: i / = CH: 321o, 314o, 3o7o; G = O: 1745, 172.0; Go / other: 133o, 1235, 1185, 1o9o, 1o5o , 99o, 935, 9oo, analysis: boron. For G ₁₄ H ₁₇ N ₅ O ₀ S: G: 43.45; H: 4.42; H: 1o.85;

S: 8.28; (found: G: 42.98; H: 4.48; N: 10.76;

S: 8th, 57; 1 - (2,3,5-'l ¹ ri-O-acetyl-ß-1) -ribof uranosyl) -2-thiouracil,

BAD OBIGINAU 2 0 9 8 2 9/1156

1 ~ (2,3,5-'i ^l ri-0-acetyl-bi; -ri'boiuranoByl) -2-triiotliyiniri, 1- (3f 5-l'i-0-acetyl-2-deoxy-ß- ! ü-threopentoiuranüsyl) -2-tiiiotJiymir ¹ DZVJ.

1- (2,5-iJi-ü-acetyl-5-deoxy-ö-JJ-tlireopentofuranosyl) -2-tliiaiiracil.

Example 9

Repeating the procedure of Example 4, 'i'eil 0, but using i ^J rO | iorisäureanhydrid, Butters Jaireanhydrid, benzoic anhydride or AnisGäuretinh'ydrid instead' of the acetic anhydride, thus obtained the compounds 1- (5,5-- ui-0-propionyl-2-deoxy-ß-; L) -erythropentoi'uranosyl) -2-thiothymine, 1- (3,5-l>iU-butyryl-2-deoxy-j'b-jj-exythropentofui^> anosyl) -2-tniothymine, 1- (3, ί? -iJi-ί> -ΐ) en2; oyl-2-deoxy-J3-l) -erytiαropentoί'uranosyl) -2-thiothymine and 1- (3.5 -Bi-U-anisoyl-2-deoxy-i3-li-erythropentoi (iranosyl) -2-thiothymine.

Example 1o

l'eil A.

"If the procedure of Example 3 is repeated, however using the starting materials 1 - (ß-D-iti bof uranosyl) -5-ÄLhyluracil, 1 - (ß-jj-iiibof urano syl) -5 -but ylurac il, 1- (ß-ij-ii.ibof uranosyl) -5-hydroxymethyluracil, 1- (ß-D-iiibo ± "uranosyl) - ^ - methoxymethyluracil, i-Cß-JJ-iiibofuranosylJ - ^ - acetoxymethyluracil or 1 "(ß_U_Hibofuranosyl) -5-äthoxymethyluracil, then one obtains the

BAD ORIGINAL 209829/1156

Links

1- (2,3-0-isopropylidene-ß-l) = ribof uranosyl) ~ 5-äthyru.racil _?

■ \ _ (2,3-U-isopropylidene-ß-iJ-ribofuranosyl) -5-butyluracil, 1- (2,3-U-isopropylidene-BO-ri "bofurarLOsyl) -t) -hydroxynietliyluracil _?

1- (2,3-0-Isopropylidene-β-.u-ri'bofuranosyl) -5-met] ioxyraethyluracil, 1- (2,3-Ü-Isoprop3rliden-β-lJ-ril3of uranosyl) -5-acetoxymethyl -uracil Td betw.

1 - (2, SO-Isopropyliden-B-iJ-rilDof uranosyl) -5-ethoxy-diethyluracil _o

Part Bo

If the procedure of Example 3, part B, is repeated, however

1- (2,3-u-Isopropylidene-ß-1) -ri "bo ± 'uranosyl) -5-äthyluracil, 1- (2,3-0-isopropylidene-ß-: u-ri'bofuranosyl) -5-'butyluracil, 1- (2,3-U-Isopropyliden-ß-U-ri "bof uranosyl) - ^ - hydroxyme thy luracil, 1- (2,3-ü-isopropylidene-ß-D-ri "boi'uranosyl) -5-niethoxymethyluracil, 1- (2,3-ü-Isopropylidene-β-D-ribofuranosyl) -5-acetoxymethyluracil

1- (2,3-0-Isopropylidene-ß-1) -ri'boi'uranosyl) -5-ethoxymethyluracil, this is how you get the connections

1- (5-Ü-Tosyl-2,3-0-isopropylidene-b-lJ-ri'bof uranosyl) -!? - ethyluracil,

1 - (5-0-To syl-2,3 -ü-isopropylidene-ß-aj-riTDoi uranosyl) -! 5-butyluracil,

1- (5-O-'i'osyl-2,3-0-isopropylitien-ß ~ D-ribofuranosyl) -5-hyclroxymethyluracil,

1- (5-0-Tosyl-2,3-0-isopropylidene-β-'Jj-ribofuranosyl) -5-methoxyme thyluracil,

1- (5-0-Tosyl-2., 3-O-isoproxDylidene-ß-ii-ri'bo ± ^> uranosyl) -5-acetoxymethyluracil or

1- (5-0-To syl-2,3-0-isopr opyliden-ß-li-ribof'ui "anosyl) -5-ethox37 · - me tliy lur ac il.

BATH ORIGINAL

209829/1156 ~~

- 4ο -

Part C.

If the procedure of Example 3, Part C is repeated, but using 1_ (5-υ-ιΌ syl-2,3 - ^ - isopropylidene-ß-E-ribofuranosyl) -5-ethyluracil, 1- (5-ü- Tosyl-2,3-U-isopropylidene-β-jü-ribofuranosyl) -i? -T) utyluracil, 1- (5-u- '±' osyl-2,5 - ^ - isopropylidene-β-l) -ri 'bofuranosyl) -5-HYDX * oxy · - methyluracil, 1- (5-0-'i ⁱ osyl-2.5-0-isopropylidene-.beta.-D-ri'bofuranosyl) -5-niethoxymethyluracil, 1- (5 -0- ⁱ i ⁱ osyl-2,3-U-isopropylidene-ß-Jj-ril3ofuranosyl) -5-aGetoxymethyluracil or - | _ (5_ (j_'l ^l osyl-2,3-0-isopropylidene-ß-JJ -ril) ofuranosyl) -5-ethoxymethyluracil, the compounds 2.5 ^I -anhydro-1- (2,3 - ^ - isopropylidene-β-1) -ri'boi'uranosyl) -5-a'Lhyluracil are obtained , 2,5'-anhydro-1- (2,3-U-isopropylidene-β-D-ri "bofuranosyl) -5-t> utyluracil, 2,5'-anhydro-1 - (2,3-0 -is <^ opyliden-ß-i> -ribofuranosyl) -5-hydroxymethyluracil, 2,5'-anhydro-1- (2,3-U-isopropylidene-ß-l) -ri'boi ^> uranosyl) -5- methoxyme thyluracil, 2,5'-arihydro-1- (2,3-U-isopropylidene-ß-i / -riTDOfuranosyl) -5-acetoxymethyluracil or 2,5'-anhydro- 1- (2, ^ -U-isopropylidene-β-II-ribofuranosyl) -5-ethoxy- • methyluracil.

Part D.

one repeats the Veri'a.hren of Example 3, Part D, but with 2,5 '-Anhydro-1 - (2,3- <J-isopropylidene-ß-l) -rite) Of uranosyl) -5-:

äthylui-acil, 2.5 ^! -Anhydro-1- (2,3-0-isopropylidene-ß-i> -ribofuranosyl) -5-butyluracil,

0 9 8 2 9/1156 BAD ORiQINAL

2,5 ^I- anhydro-1- (2,3-u-isopropylidene-ß-3J-ribof'uranosyl) -!? - hydroxymethyluracil,

2.5 ^l -AnhydrQ-1- (2,3-G-isopropylidene-ß-l) -ribofuranosyl) -5-methoxymethyluracil,

2,5'-AnIydro-1- (2,5-0-isopropylidene-β-iJ-ri "bofuranosyl) -5-acetoxymethyluracil or

2.5 ^l anhydro-1- (2,3-0-isopropylidene-β-I) -ri'bofuranosyl) -!? - ethoxymethyruracil, the compounds 1- (2,3-0-isopropylidene-β -D-ri "bof-uranosyl) -5-ätliyl-2-ü-methyluracil,

1- (2,3-ü-Isopropylidene-ß-lJ-ribo £ uranosyl) -5-butyl-2-0 ~ methyluracil,

1-r (2,3-u-Isopropylidene-ß-jj-rilDoi'uranosyl) -5-hydroxymethyl-2-u-methyluraci1,

1- (2,3-0-Isopropylidene-β-U-ri'bofuranosyl) -5-methoxymethyl-2-O-methyluracil.

1- (2-, 3-t) -Isopropylidene-β-D-rit) oiuranosyl) -5-acetoxyiiiethyl-2-0-methyluracil "respectively.

1- ^ 2,3-0-Isopropylidene-β-D-ri'boi'uranosyl) -i3-ethoxymethyl-2 ~ 0-methyluracil.

Part E. - '

If the procedure of Example 3, Part E is repeated, however with

1- (2,3-ü-isopropylidene-ß-i) -riTDOfuranosyl) -5-ethyl-2-ü-methyluracil,

■ 1- (2,3-U-isopropylidene-ß-D-ri'bofuranosyl) -5-l3utyr-2-u-methyluracil,
1- (2,3-ü-Isofjropyliden-ß-i) -ri'bofuranosyl) -5-hydroxymethyl-2-0-methylura eil,

1- (2,3-0-lijoijropyliden-ß-iJ-ri'bO ± ^I uranosyl) -5-methoxymethyl-2-u-methylui'acil,

1 - (2,3-u-isopropylidene-β-D-ri "bof uranosyl) -5-acet oxymethyl-2-O-methyluracil or

H2,3-Ü- ± sopropylidene-β-jj-ribofuranosyl) -5-ethoxy-methyl-2-0-methyluracil, the compounds 1- (2,3-Ü-isopropylidene-β-JJ-ribofuranosyl) -5 are obtained -äthyl-2-thiotii * acil, 1- (2,3-iJ-isopropylidene-ß-jJ-ribo ± "uranosyl) -5-" butyl-2-th.iouracil, 1- (2,3- ⁽ - ⁾ -Isopropylidene-ß-D-ribofuranosyl) -5-hyaroxymethyl-2-thiouracil,

1- (2,5-ü-Isopropyliaen-ß-! Ü-ribofuranosyl) -5-uiethoxyme "bhyl-2'-thiouracil,

1- (2,3-U-Isopropylidene-J3-D-ribo ± uranosyl) -5-acetoxymethyl-2-thiouracil respectively.

1- (2,5-0-13opropylidene-ß-.ü-riboi'-uranosyl) -5-etb.oxymeth.yl-2-thiioaracil.

ieil l ¹ .

If the "procedure of Example 3, part jj 'is repeated, but with 1- (2,3-0-Isopi-opylid.en-ß-iJ-ribofuranosyl) - | 5-ethyl-2-thiouracil, 1- (2 , 3- ⁽ I-isopropylidene-β-l! -Ribofuranosyl) -5-t> utyl-2- "biouracil, 1- (2,3-u-isopropylidene-β-D-ribofuranosyl) -5-hydroxymethyl-2 -thiouracilj

1- (2,3-ü-Isopi ^> opyliden-ß-li-ribof'uranosyl) -5-methoxyraetliyl-2-thiouracil,

1- (2,3-ü-Isopropylidene-β-D-ribofuranosyl) -5-acetoxymethyl-2-thiouracil or

1- (2,3-ü-Isopropylidene-β-D ~ ribofuranosyl) -5-ethoxymethyl-2-thiouracil, the compounds -] _ ß_jj_Kibofuranosyl-5-ethyl-2-thiouracil, 1-ß-1J-ribofuranosyl are obtained -3-butyl-2-thiouracil _l 1-ß-I) -ribofuranosyl-3-hydroxymethyl-2-thiouracil, 1-.beta.-D-Kibo ± 'uranosyl-5-methoxymethyl-2-thiouracil, 1-ß-D -Kibofuranosyl-5-acetoxymethyl-2-thiouracil or 1-β-D-kibofuranosyl-5-ethoxymethyl-2-thiouracil.

209829/1158

Example 11

Preparation of 1- (2-deoxy-ß-D-erythropentofuranosyl) -2-thiothymine.

Part A: 1- (5-0-t'osyl-2-deoxy-ß-D-erythropentofuranosyl) -th; yTnin

A solution of 2.36 g (12.0 millimoles) of p-toluenesulfonyl chloride in 12 ml of pyridine is slowly added to a solution of 2.0 (10.0 millimoles) 1- (2-jeoxy-13-] I-erythropentofuranosyl with stirring ) -thymine in 12 ml of pyridine "was ^{added dropwise at 0 0} O. Moisture was excluded. The reaction vessel was tightly closed and left to stand at 3 to 5 G for 17 hours.

Bach added little crushed & is chilled for another hour. Then the cold reaction mixture was in about 75 ml pounded ice and poured water, then left the diluted The aqueous mixture was heated to 25 ° C., whereupon it was extracted 3 times with 50 ml of chloroform each time. The chloroform extracts were combined and washed 3 times with saturated aqueous sodium bicarbonate solution and 1 χ with water. then Filter the chloroform solution with anhydrous sodium sulfate dried. After evaporation of the chloroform under reduced pressure The residue was azeotroped with absolute ethanol under reduced pressure, and the pressure the resulting white solid became from 95tiger aqueous Ethanol crystallized using decolorizing charcoal, followed by drying at reduced pressure and 40C v / urde. This gave 2.22 g of 1- (5-u-tosyl-2-deoxy-ß-D-erythropentofuranosyl) thymine from melting point 17o to 171.5 ° C. UV absorption:

223, 265, 273 (Sch) (6 = 17ooo, 98oo, 885o)

Characteristic. lit absorption bands: (cm ") Uri / Nh7 = CM: 337o, 516o, 31oo, 3040; C = ü / C = C: 172o, 1655, 1595,

209 8 29/1156

-SOgü / C-ü / others: 156ο, 1275, 119ο, 1175, 1ο95, 1ο75, 925, 92ο; pJ-CH / Others: 85ο, 82ο.

Analysis: Ber. for G ₁₇ H ₂₀ N ₂ O ₇ S: C: 51.56; H: 5, ο9; N: 7, ο7;

S: 8.1ο. Found: C: 51.83; H: 5, ο5; N: 6.69;

S: 8.92.

Part B: 2.5 ^f -Anhydro-1- (2-deoxy ~ ß-D-erythropentofuranosyl) -

thymine.

Mn mixture of 1.98 g (5.0 millimoles) 1- (5-0-tosyl-2-deoxyß-D-ery.thropentofuranosyl) thymine (see Part A), 60 ml of dimethylformamide and 5 g of sodium bicarbonate was added with stirring Heated in a nitrogen atmosphere * for 2 hours on a bath with a bath temperature of 122 to 154 C. Then it became The reaction mixture was cooled to about 25 O, filtered and the filter was washed with dimethylformamide. After uniting The dirnethylformamide wuxed out of the filtrate and washing solutions evaporated under reduced pressure and the residue was azeotroped under reduced pressure with 95% aqueous ethanol subject. The residue obtained was redissolved in 95 / ^ strength aqueous ethanol and the 5 g of silica gel was added to the solution. After removing the aqueous ethanol under reduced pressure, it became dry G-emically applied to a chromatography column with 150 g of silica gel. The column was developed with solvent C and 750 ml fluid was collected. After removing the solvent G at reduced pressure, 660 mg of 2,5'-anhydro-1- (2-deoxy-β-D-erythropentofuranosyl) thymine were obtained from the melting point 193-197 ° C.

^ Ä

UY absorption: ^ ^ ÄtüH _{u &}

Part C: 1- (2-Deoxy-β-D-erythropentofuranosyl) -2-0-methylthymine.

209829/1156

Line solution of 7.96 g (31st, 2 millimoles) of 2,5'-anhydro-1- (2-deoxy-β-D-erythropentofuranosyl) -thyrnine (see Part B), 76 ml .dimethylformamide, 76o ml of absolute methanol and 765 mg (14.15 millimoles) sodium methylate was sealed in a Left flask for 19 hours. Then the solution is neutralized by the slow dropwise addition of glacial acetic acid. Small amounts of a flaky precipitate were filtered off and the filtrate was removed under reduced pressure freed from the solvents. The residue was at reduced Pressure with absolute ethanol an azeotrope treatment

/became

subjected, thereby 8.56 g of 1- (2-Deoxy-ß-D-erythropentoxuranosyl) · 2-ü-methylthymine, which by small amounts of sodium acetate contaminated, but for further reaction with hydrogen sulfide was suitable to receive.

Example 12

Part A.

If the procedure of Example 11, Part A is repeated, but using the starting materials 1- (2-deoxy-ß-D-erythropentofuranosyl) -6-azauracil, 1- (2-deoxy-ß-D-erythropentofuranosyl) -5- methyl-6-azauracil (for the preparation of both compounds, see Fliml, Pyystas, and Sorm, QöLlection Czechoslov. Chem. Gommun. 28, pp. 2588-2596 (1963)), 1-ß-D-arabinjäiofuranosyl-6-azauracil or 1 -ß-D-arabinofuranosyl-5-methyl-6-azauracil, the compounds 1- (5-Ü-tosyl-2-deoxy-ß-I) -erythropentofuranosyl) -6-azauracil, 1- (5- U-! I ^l osyl-2-deoxy-ß-D-erythropent of uranosyl) -5-methyl-6-azauracil,

! - (^ - D-l'osyl-ß-D-arabinofuranosyl) -6-azauracil or 1-jj ^ -. 0- ^r i'osyl-iJ-D-arabinofuranosyl) -5-methyl-6-azauracil .

209829/1156

'JJeil- B.

"If you repeat the procedure of Example 11, Part B, however

1 - (5-0-Tosyl-2-deoxy-ß-I) -erythropentofuranosyl) -6-azauracil, 1- (5-U-! I ^l osyl-2i-deoxy-ß-iJ-erythropentofuranosyl) -5- methyl-6-azauracil,

1- (5-0-l'osyl-β-I) -ara'binofuranosyl) -6-azauracil or i-CS-O-Tosyl-ß-D-arabinofuranosyli-S-niethyl-6-azauracil, this is how you get the connections

2,5'-Anhydr0-1- (2-deoxy-ß-iJ-erythropentofuranosyl) -6-azauracil, 2,5'-anhydro-1- (2-deoxy-ß-jj-erythropentofuranosyl) -5-niethyl-6-azauracil,

2,5'-anhydro-i-ß-Ii-arabinoiuranosyl-ö-azauracil or 2,5'-anhydro-1-ß-U-arabinofuranosyl-S-methyl-δ-azauracil.

Part C.

If the procedure of Example 11, Part O is repeated, however

2,5'-anhydro-1- (2-deoxy-ß-U-erythropentofuranosyl) -6-azauracil, 2,5'-Anhyäro-1- (2-deoxy-ß-U-erythropentofuranosyl) -5-methyl-6-azauracil,

2,5'-anhydro-1-β-I) -arabinofuranosyl) -6-azauracil or 2,5'-anhydro-1-ß-U-arabinofuranosyl-S-methyl-b-azauracil, this is how you get the connections

1- (2-Deoxy-ß-D-erythropentofuranosyl) -2-O-methyl-6-azauracil, 1- (2-Deoxy-ß-D-erythropentofuranosyl) -2-ü-methyl-5-raethyl-6-azauracil,

-] "ß_D_Arabinofuranosyl-2-0-methyl-6-azauracil or 1-β-D-arabinofuranosyl-2-0-methyl-5-methyl-6-azauracil.

Part D.

If the procedure of Example 4, Part B is repeated, but with 1- (2- ±) eoxy-ß-l> -erythropent of uranosyl) -2-ü-methyl-b-azauracil,

209829/1156

1- (2-Oeoxy-ß-lJ-erythropentof uranosyl) -2 ~ 0-methyl-5-methyl-> 6-azauracil,

1-ß-l) -arabinofuranosyl-2-0-methyl-6-azauracil or 1-ß-D-Ara "binofuranosyl-2-U-methyl-3-inet] iyl-6-azauracil, this is how you get the connections

1 '- (2-üeoxy-ß-D-erythropentofuranosyl) -Z-thio-G-azauracil, 1- (2-I) eoxy-ß-I) -erythropentof uranosyl) -2-tliio-5-methyl-6-azauracil,

1-ß-jj-Ara "binofuranosyl-2-thio-6-azauracil or 1-ß-D-Ai-abinof "uranosyl-5-niethyl-2-thio-6-azauracil.

Rope i ..

the procedure of Example 4, Part C, is repeated, however

1 - (2-l) eoxy-ß-D-erythr opentof ruanosyl) -2-thio-6-azauracil, 1- (2-Jjeoxy-ß-D-erythropentofuranosyl) -2-thio-5-methyl-6-azauracil,

i-ß-L-arabinofuranosyl ^ -thio-G-azauracil, or 1-ß-l) -arabinofuranosyl-5-methyl ~ 2-thio-6-azauracil, this is how you get the connections

1 - (3 1 5-i) i-0-acetyl-2-deoxy-ß-D-erythropent of uranosyl) -2-thio-6-s, zauracil,

1 -) 5,5-iJi-O-acetyl-2-de oxy-ß-l> -erythr opentof uranosyl) -5- methyl-2-thio-6-aza-uracil,

1 - (2,3-5-i'i'i-O-acetyl-β-D-arabinof uranosyl) -2-thio-6-azauracil bzv /.

1 - (2,3,5-l'ri-0-acetyl-β-l) -arabinof uranosyl) -S-6-azauracil.

2098 29/1 15 6

Claims (22)

Claims c h e y%. 1 ~ (2- or 5-I) eoxy-B -]> - pentofuranosyl) -2-thiouracils iind 1-ß-D-pentofuranosyl-2-thio-6-aza-uracils of the formula YCH Ii HN C-M I II ³ ^ _n in the M hydrogen, a lower alkyl radical with 1 to carbon atoms, a hydroxy-lower alkyl radical with 1 to. 3 carbon atoms, a lower alkoxymethyl radical with 2 to 5 carbon atoms or a lower alkanecylic acid ester of the hydroxy-lower alkyl radical with a total of 3 to 6 carbon atoms, X the group = CH- or = N-, Y a hydroxyl group, a lower alkanoyl radical with 1 to 4 carbon atoms or an aroyl radical with 7 "to 8 carbon atoms? Q. Y) C substance atoms and Z, Z, Z, Z independently of one another 209829/1156 represent a hydrogen atom, a hydroxyl group, a lower alkanoyl radical with 1 to 4 carbon atoms or an aroyl radical with 7 "to 8 carbon atoms, provided that at least one of the substituents, but not more than two at the same time are hydroxyl, lower-alkanoyl or aroyl groups , with the further proviso that Z and Z ^c , Z ^a and Z ^c , Z and Z ^a and Z and Z cannot both be hydroxyl groups or lower alkanoyl groups when X is a = GH-u group. 2. A compound according to claim 1, characterized in that X is a = Cti group. 3. A compound according to claim 2, characterized in that the deoxy-ß-i) pentofuranosyl radical is a 2-deoxy-ß-li-pentofuranosyl radical is. 4 «connection according to claim 5, characterized in that M is lower alkyl. 5. 1- (2-L) eoxy-β-I) -erythropentofuranosyl) -2-thiothymine. 6. A compound according to claim 4, characterized in that Z ^a and Y are lower alkanoyl radicals. 7 · 1- (3,5-x) i-0-acetyl-2-deoxy-β-D-erythropentofuranosyl) -2-thiothymine. 8. 1- (2-Deoxy-β-D-threopentofuranosyl) -2-thiothymine. 9. A compound according to claim 3, characterized in that M is a hydrogen atom. 10. 1- (2-eoxy-β-U-erythropentofuranosyl) -2-thiouracil. 2 0 9 8 2 9/1158 11. A compound according to claim 9, characterized in that Z and Y are lower alkanoyl radicals. 12. 1- (3,5-M-u-acetyl-2-deoxy-β-U-erythropentofuranosyl) -2-thiouracil. 13. 1- (3-Jeoxy-β-lJ-threopentofuranosyl) '- 2-thiouracil. 14. A compound according to claim 1, characterized in that X = M-. 15. A compound according to claim 14, characterized in that M is a hydrogen atom. 16. 1-β-L) -li.ibo ± 'uranosyl-2-thio-6-azauracil. 17.1 - (2,3 j 5-'l'ri-ü-acetyl-ß-ii-ribofurano syl) -2-thio-b-azauracil, 18. Process for the preparation of i-ß-D-pentofuranosyl-2-thiouracils or - ^ - thio-ö-azauracils, characterized in that "one (2.5 ^t -anhydro-1-ß-I) -pentofuranosyl ) -uracil- or -ö-azauracil-nueleosid, in which M, Z, Z ^a , Z and Z ^{c are} non-reacting substituents, reacted with methanol in the presence of an alkali metal alkoxide to "form" the corresponding 1-ß-D-pentofuranosyl -2-Ü-riiethyluracils or -2-O-methyl-6-azauracils and this treated with hydrogen sulfide. 19. The method according to claim 18, characterized in that a 2- or 3-deoxy-entofuranosyl group is present. 20. The method according to claim 18 or 19, characterized in that that sodium methylate is used. 21. Pharmaceutical preparations containing a pharmaceutically acceptable carrier and an active ingredient of the i'ormel 209829/1156 YCH ₂ in which M is hydrogen, a lower alkyl radical with Ί. up to 4 carbon atoms, a hydroxy-lower alkyl radical with 1 to carbon atoms, a lower alkoxymethyl radical with 2 to carbon atoms or a lower alkanecarboxylic acid ester of the hydroxy-lower alkyl radical with a total of 3 t> is 6 carbon atoms, X the grouping = CH- or = N-, Y is a hydroxyl group, a lower alkanoyl group with 1 to 4 carbon atoms or an aroyl group with 7 to 8 carbon atoms and Z, Z ^a , Z and Z ^c independently of one another a hydrogen atom, a hydroxyl group, a lower alkanoyl group with 1 to 4 carbon atoms or an aroyl residue with 7 Ms 8
Represent carbon atoms, with the proviso that at least one, but not more than two of the substituents
are at the same time hydroxyl, lower-alkanoyl or aroyl groups, with the further proviso that Z and Z, Z ^a and Z ^c and 'bc ' Z and Z are not both hydroxyl groups or lower alkanoyl groups can be when X is a = CH group. 2098 29/1156 22. Preparation according to claim 21, containing as active ingredient 1 -BD-HiTDOfuranosyl-2-thiouracil, 1-ß-D-ribofuranosyl-2-thio.6-azauracil, 1- (2,3,5-l ¹ ri-0 -acetyl-β-I) -ribofuranosyl) -2-thio-6-azauracil, 1- (2-I> eoxy-β-D-erythropentofuranosyl) -2-thiothymine, 1- (2-deoxy-β-D -erythropentofuranosyl) -2-thiouracil, 1 r (2-Ueoxy-.beta.-D-threopentofuranosyl) -2-thiothymine, 1- (3-deoxy-.beta.-D-threopentofuranosyl) -2-thiouracil, 1- (3,5 -Di-O-acetyl-2-deoxy-ß-u-erythropentofuranosyl) -2-thiouracil or 1- (3,5-Ui-0-acetyl-2-deoxy-ß-D-erythropentofuranosyl) -2-thiothymine. For: The Upjohn Company Kalamazoo, Mich., V.St.A. (Lawyer) 209829/1156