HUT75338A - Adenosine derivatives, their preparation and use, and pharmaceutical preparations containing them - Google Patents

Description

The pharmaceutical compositions of the present invention contain, in addition to pharmacologically acceptable carriers or diluents, an effective amount of a compound of formula (Ia) in the treatment of the present invention, such as relieving pain, relieving high blood pressure, and expanding the blood pressure. . patients with high blood lipid levels, hypertriglyceridemia, and low HDL cholesterol, as well as type I or type II. patients with type 2 diabetes, etc. ⁽ - < tb > ingesting a sufficient amount of a compound of formula (la)). The present invention provides 2 (O-alkyl) adenosine derivatives by treating adenosine with appropriate alkyl sulfates in the presence of a phase transfer catalyst.

-A- (The process according to the invention is less time consuming and costly in comparison to the prior art processes and can be used in the preparation of more pure active substances) (ι v'U

(Λ

S 1 'η' ι L _f

h.'ü V. ·> 6, ... _Ί ff t. (Li./ <tí {· _M ί, n {., ·· k V _v . [.

Ti '•' / 04 Z ^C

Represented by Danubia Patent and Trademark Office Ltd., Budapest

Adenosine derivatives, their preparation and use, and pharmaceutical compositions containing them

SANDOZ LTD., Basel,

inventors:

FOZARD John R.,

GADIENT Fulvio,

MANGOLD Bonnié L.K., KAPA Prasad Koteswara, PRASHAD Mahavir,

Switzerland

Hegenheim, FR Birsfelden, CH Mendham, N.J., US Parsippany, N.J., US

Hopatcong, N.J., US

Date of International Application: 09.05.1995

PRIORITY: 1994. 05th 10th (9409324.2, GB) 1994th 05th 26th (08/249914, US) 1994. 08 18th (9416693.1, GB)

International Application Number: PCT / US95 / 05802 International Publication Number: WO 95/30683

84569-2703-PT / KmO <... :

The present invention relates to adenosine derivatives, to processes for their preparation and to their use, and to pharmaceutical compositions containing such derivatives. In particular, the present invention provides new uses for A1 receptor agonists.

In particular, the present invention relates to a novel use of compounds of formula I wherein R is hydrogen, C 1-4 alkyl, allyl, metallyl, straight or branched C 3-7 alkynyl, C 3-8 cycloalkyl, hydroxy, (C 4 -C 8) cycloalkyl, including phenyl, phenyl substituted with one or more substituents independently selected from the group consisting of halogen, C 1-4 alkyl, C 1-4 alkoxy, trifluoromethyl and trifluoromethyl; phenyl, (C 1 -C 4) alkyl, including the limit values on the phenyl ring, having from 9 to 35 halogen, (C 1 -C 4) -alkyl, (C 1 -C 4) -alkoxy or trifluoromethyl (singly or independently) - doubly substituted phenyl (C1-C4) alkyl; C 1-4 alkyl substituted with at least one hydroxy group or at least two phenyl groups; a bicycloalkyl group such as endo or exo-bicyclo [2.2. l] heptyl -; naphthyl (C 1 -C 4) alkyl, acenaphthylenyl (C 1 -C 4) alkyl; or something like that.

- a group of 3, wherein Z is hydrogen, hydroxy or C 1-4 alkoxy;

- Q is hydrogen or hydroxy;

- A represents a methylene group, an oxygen atom, a sulfur atom or a direct bond;

- Y is a direct bond or - (CH ₂ ) _n - wherein n is 1, 2 or 3;

and the dotted line in formula (A) represents an optional bond; and

- R ₂ is hydrogen, (C 1 -C 4) -alkyl, amino, (C 3 -C 5) -cycloalkyl, or within the limits - from 9 to 35 halogen; and

- R ₃ is C 1-4 alkyl.

The compounds of formula (I) are believed to - for example, the _R: site other than hydroxyl cycloalkyl group-containing substituents - are known and have been disclosed [for example, the No. 2 226 027. United Kingdom Patent Publication No. 378,518; and 269,574. European Patent Applications, and U.S. Patent No. 4,843,066; and U.S. Patent Nos. 4,985,049; U.S. Patents].

In the formula (I), the tetrahydrofuran ring substituents at the 2'-, 3'-, 4'- and 5'-positions have the same configuration as the corresponding substituents on the adenosine molecule.

A class of compounds of formula I is represented by β · i · i · i ·

- 4 (RJ is a substituent other than hydroxycycloalkyl). Another group of compounds of formula I is Rx is hydroxy (C4-C8) cycloalkyl.

Preferably, the compound of formula (I) is 6-cyclohexyl-2 '- (O-methyl) -adenosine, hereinafter referred to as compound (M).

In the preferred compounds of formula (I), the A? -Adenosine activity can usually be determined as follows.

Affinity for adenosine receptors

Porcine corpus striatum membranes were prepared for assay as previously described by H. Bruns et al., Molecular Pharmacology, 29, 331-344 (1986).

³ H-NECA, a non-selective adenosine receptor antagonist, is used to label both the A _T receptor and the A ₂ receptor. IC ₅₀ values are determined from the displacement curves of the non-linear weighted curve fitted to the Langmuir equation and the calculated pK _D values.

Spreadsheet

Affinity of adenosine receptor ligands for A _x and A ₂ receptors

Αχ (K _D ; nM) A ₂ (K _D ; nM) Αχ: Selectivity ₂ CPA 0.74 ± 0.08 930 ± 110 1260 6 CV 1808 2460 ± 757 269 ± 70 0.1 5 CGS 21680 4360 ± 1080 18.6 ± 4 0, 004 z Compound (M) 1.5 H ₂ O 23 ± 2 24500 ± 5160 1090 5

- 5 CPA = cyclopentyladenosine

CV 1808 = 2-Phenylamino-adenosine [Carbohydrates, 81,

No. 91898 No. K. review (1974)]

CGS 21680 = 2- [p- (2-Carboxyethyl) phenethylamino] -5'-N-ethylcarboxamidoadenosine [FASEB J., 3, (4),

4770th-4773rd s. abstracts (1989)]

According to the literature, CPA has been shown to be an effective and highly selective displacement agent for A 1 receptor binding components, CV 1808 is a relatively weak but selective A ₂ receptor ligand, and CGS 21680 is very effective and selective for the A ₂ receptor.

Compound (M), in the form of 1.5 crystalline water, exhibits strong affinity and high selectivity for the A ₂ receptor as opposed to the A ₂ receptor.

Known compounds of formula I are known as antihypertensive agents and coronary vasodilators.

It is also known that the compounds of formula I protect the endothelium of blood vessels by preventing platelet adhesion and activating white blood cells. These compounds also lower blood lipid levels. In addition, the compounds of formula (I) provide protection against diseases caused by hypertension such as congestive heart failure, myocardial infarction or sudden cardiac death and renal failure. and 269,574. European Publication Documents].

It is also known that said compounds may be useful in the treatment of neurodegenerative disorders, peripheral nerve disorders such as diabetes mellitus, peripheral vascular disorders and / or neurodegeneration-related disorders, hypertriglyceridemia, high free fatty acid levels, for the treatment of type 1 or type 2 diabetes, including non-insulin dependent diabetes mellitus, and cardiac arrhythmias, in particular for the development of hypertrophy and ventricular myocardial infarction.

In our experience, the compounds of formula (I) are particularly useful as anesthetics, for example in the treatment of pain, such as acute or chronic pain.

The anesthetic activity of the compounds of Formula I is demonstrated by their analgesic activity in standard animal studies using, for example, anti-inflammatory or neuropathic models, such as, for example, prolonged inflammation of the ) and (b) tests] and in the reduction of prolonged heat-induced pain sensation (thermal hyperalgesia) [test c)].

Increased pain sensation caused by inflammation

a) Test: induction of increased pain sensation with Freund's enhancer

One knee joint of rats is injected with 100 microliters of Freund's complete booster. The load on rats in the injected leg is reduced and remains low for up to 5 days. This mechanism of mechanical hyperalgesia is influenced by NSAIDs and opiates. The compounds of formula (I) are administered at a dose of from about 3 to about 60 pg / kg of body weight by injection or, preferably, orally. On the injected side, we measure the increase in endurance to determine the progressive withdrawal of hyperalgesia.

Particularly noteworthy is the activity of Compound (M), which has a duration of action of about 1 hour when administered orally at a dose of about 3 to 60 pg / kg body weight. Given that no significant differences were observed in the response of the animals at doses of 3, 30 and 60 pg / kg body weight, it can be concluded that

Maximum effect was achieved in the range of 3-30 pg / kg body weight.

b) Test: Induction of mechanical hyperalgesia with turpentine

Locally injected turpentine / paraffin injected into the sole of the left hind paw of rats induces inflammation around the injection site and reduces the protective threshold (suppression threshold 340 g) for mechanical stimulation (pressure on the paw). Compounds of formula I, administered orally or subcutaneously 3 days after injection, are effective at a dose of 1 to 100 pg / kg body weight. New Thresholds at 1 and 3 Hours «4 4 4 * 4« «« t

- 8 set.

Compound (M), when administered orally at doses of 30 and 60 µg / kg body weight, shows significant activity. The effect is maximal at a dose of 30 gg / kg body weight. The morphine ED ₅₀ value for this test is 1.2 mg / kg body weight when injected subcutaneously.

Neuropathic hyperalgesia

c) Test: Examination of Neuropathic Thermal Hyperalgesia according to the principles of Z. Seltzer et al., Pain, 43: 205-218 (1990).

Partial ligation of the sciatic nerve removes nerve fibers from the nerve network of the rat paw. Partially deprived of nerve fibers, without triggering reflex separation, mechanical stimuli and heat stimuli develop increased pain and allodynia. The animals were placed in a perspex box on a thin glass slide. The soles of the paws of the animals are incrementally stimulated with increasing heat and the time until the paw is retracted is measured. The compounds of formula I are effective 12 to 15 days after nerve dressing, when administered subcutaneously or, preferably, orally, in a dosage of about 1 to 100 µg / kg body weight. Compound (M) is particularly effective against increased pain sensation when exposed to heat. Compound (M) exhibits significant activity at doses of 30 to 60 µg / kg body weight, with maximal effect at 30 µg / kg body weight. For oral administration, the ED ₅₀ is about 60 µg / kg body weight. In this test, the ED ₅₀ value for morphine - approximately subcutaneous injection

mg / kg body weight.

Clinical trials

Clinical trials may be performed as described in the following section.

Compounds of general formula (I), in particular Compound (M), are injected intravenously in a dose of 0.02 to 5 mg in a subject suffering from pain, post operative pain or post-ring neuralgia. We can observe a reduction in pain.

Thus, the compounds of the invention may be used as anesthetics, for example in the treatment of acute or chronic pain such as chronic neuropathic pain.

Therefore, one result of the present invention is to provide a method of alleviating pain, comprising administering to a subject in need thereof a sufficient amount of a compound of formula (I) as hereinbefore defined for its therapeutic effect.

The present invention makes it possible to use the compounds of formula (I) as hereinbefore defined as anesthetics for the preparation of medicaments for the treatment of pain.

A further result of the present invention is the availability of the compounds of formula (I) as previously defined for analgesia.

It is also within the scope of the present invention to provide analgesic compositions which are pharmaceutically acceptable carriers or

Containing 10 of the compounds of formula I as defined above in addition to diluents.

Indications for the use of the compounds of the formula I: pains such as acute pain associated with tissue damage and inflammation (such as post-operative pains, burning pains, pain-related pains, etc.), chronic inflammatory pains (such as arthritis) and chronic neuropathic pain (e.g. such as neuropathy associated with diabetes, neuralgia after ringworm, multiple sclerosis, post-amputation pain, etc.).

Of course, the dosage of the compounds used in the above indicated uses of the invention will depend upon the active ingredient employed, the severity of the disease, the subject being treated, the patient's body weight, the mode of administration and the relative potency of the active ingredient. In general, however, satisfactory results can be obtained when treating animals with doses ranging from about 1 to 100 µg / kg body weight, such as 3 to 60 µg / kg body weight, such as 10 to 60 µg / kg body weight, three times daily. In the treatment of larger mammals, such as humans, the recommended daily dose is about 0.1 to 10 mg, which, when properly administered, is administered in unit doses of about 0.02 to 5 mg more than once a day, for example 2, 3, 4, 5 or 6 times - it can be delivered to the body, but once or twice daily it is advisable.

With test a) described above, the ED _{50 of} ibuprofen orally administered NSAID ibuprofen is approximately 4 mg / kg body weight and that of indomethacin - also orally • ·

- 11 mg / kg body weight. Compound (M) is about 300-fold more potent. Depending on the severity of the disease (s) and the frequency of administration, the preferred dose of Compound M is about 0.1 to 10 mg / day, for example, 3 to 30 pg / kg for a 70 kg adult.

Compound (M) has been shown to have no adverse effects on the human body following single oral administration of single doses up to 5 mg.

The term "pharmacologically acceptable" as used herein is intended to be used with reference to substances that are useful in both human and veterinary medicine.

The compounds of the present invention can be formulated into ready-to-use compositions by any suitable means. Depending on the disorder to be treated, which formulation method should be chosen. It is advantageous to formulate the compositions in unit dosage form or in such a form that the patient may self-administer a single dose of the vaccine. Preferably, the compositions may be administered orally, rectally, topically, parenterally or intravenously or intramuscularly.

The compositions of the present invention may be formulated as tablets, capsules, sachets, vials, powders, granules, lozenges, suppositories, water-soluble powders or liquid preparations such as oral solutions or suspensions, or sterile solutions or suspensions for parenteral use. . Topical formulations may also be used where appropriate.

In order to maintain a constant dosage, it is preferable to administer the compositions of the invention in unit dosage form.

For oral use, unit dosage forms may be formulated as tablets and capsules containing 0.02 to 5 mg of the compounds of the present invention, which may further contain conventionally used excipients such as syrup, acacia, gelatin, sorbitol, tragacanth or polyvinylpyrrolidone. ), fillers such as lactose, sugar, corn starch, calcium phosphate, sorbitol or glycine, tabletting lubricants such as magnesium stearate, tablet disintegrants such as starch, cross-linked polyvinylpyrrolidone, sodium starch glycolate or microcrystalline cellulose; and / or pharmacologically acceptable wetting agents such as sodium lauryl sulfate.

Solid compositions for oral use may be prepared by conventional means such as mixing, filling, tabletting, and the like. In the case of formulations using a large amount of filler, repeated mixing operations may be carried out to obtain an appropriate distribution of the active ingredient.

Of course, the above operations are conventionally used in the preparation of medicaments. The tablets may be coated by methods well known in the art, and in particular by enteric coating.

Liquid formulations for oral use may include emulsions, syrups or medical drinks, but may also be formulated in a dry formulation for constitution with water or other suitable vehicle before use. These liquid formulations may contain commonly used additives such as suspending agents such as sorbitol, syrup, methylcellulose, gelatin, hydroxyethylcellulose, carboxymethylcellulose, aluminum stearate gel or hydrogenated edible fats such as emulsifiers, lecithin, sorbitan monooleate or acacia, non-aqueous vehicles which may also be edible oils, such as almond oil, fractionated coconut oil, oily esters such as glycerol esters and propylene glycol esters; or ethyl alcohol, preservatives, such as methyl p-hydroxybenzoic acid, propyl p-hydroxybenzoic acid or sorbic acid, and, if necessary, conventional flavoring or coloring agents.

Following are examples of solid compositions for oral use, such as tablets and capsules.

The tablets were compressed from capsules of the appropriate dosage strength. Compound (M) was encapsulated using size 3 yellow capsules. The composition of the 0.1mc and 5mg strength formulations is described below.

* · · ·

0.1 mg tablets / capsules 5.0 mg tablets / capsules Compound (M) 0.1 5, 0 Microcrystalline cellulose 76, 92 74.96 Water-containing lactose 87.63 * 84, 69 ' Poly (vinylpyrrolidone) 7.4 7.4 crospovidone 7.4 7.4 Magnesium stearate 0, 925 0, 925 Remaining moisture 4, 625 4, 625 Size 3 capsules 50 50 The substance filled into the capsule or tablet theoretical weight 185 185 The theoretical weight of the capsule 235 235

The amount of lactose is adjusted to compensate for the amount of moisture in the drug substance.

The compounds may also be formulated as sustained release formulations to extend the duration of action. Conventional formulations such as coated pellets or push-pull osmotic systems may be used to provide a sustained action.

For parenteral administration, unit dosage forms are formulated in a liquid dosage form using the active ingredient and a sterile vehicle. Depending on the concentration used, the active ingredient is suspended or dissolved in the vehicle. When preparing solutions, the active ingredient is dissolved in polyethylene glycol or ethanol and diluted with water for injection, then sterilized by filtration before filling into the appropriate vial or ampoule and sealing the vial or vial. Excipients such as topical anesthetics, preservatives and buffering agents should be dissolved in the vehicle. Parenteral suspensions are prepared in substantially the same manner with

Except that the active ingredient is not dissolved, but suspended in the vehicle and sterilization cannot be accomplished by filtration. The active ingredient may be sterilized by exposure to ethylene oxide prior to suspension in the sterile vehicle. It is also desirable to include a surfactant or wetting agent in the compositions to facilitate uniform distribution of the active ingredient.

The formulations may contain from about 0.1% to about 99%, preferably from about 10% to about 60%, by weight of the active ingredient, depending on the route of administration.

The compounds of formula I, as well as their hydrates, addition products and non-aqueous solvents, and salts thereof, may also be used in combination with excipients commonly used in topical formulations in the form of topical formulations.

Topical formulations include ointments, creams, lotions, impregnated dressings, gels, gel sticks, sprays and aerosols. These compositions may contain suitable customary additives, for example preservatives, solvents to facilitate the penetration of the active ingredient, and, in ointments and creams, plasticizers. They may also be compatible carriers conventionally used in the compositions, such as cream or ointment bases and, in the case of lotions, ethanol or oleyl alcohol.

Suitable creams, salts of the compounds of formula (I), their hydrates, solvents other than water and salts thereof may be prepared.

The composition of the 16 detergents, gels, ointments, sprays and aerosols corresponds to formulas conventionally known in the art. Such prescriptions are described, for example, in manuals widely used by pharmacists and cosmetologists, such as Harry Cosmeticology, published by Leonard Hill Books, Remington's Pharmaceutical Sciences, and the British Pharmacy and the United States Pharmacology.

As a result of the present invention, adenosine derivatives having the general formula (Ia) are also available

- R 4 is C 4 -C 8 cycloalkyl substituted with hydroxy;

- R ₂ is as defined above, preferably hydrogen,

C 1 -C 4 alkyl or halogen atoms, including the limit values, from 9 to 35; and

R ₃ is as defined above.

The compounds of formula (Ia) may exist in the form of enantiomers or diastereomeric mixtures.

In the formula (Ia), the limit values, including the limits, may be from 9 to 35 halogen fluorine, chlorine or bromine; C 1-4 alkyl is methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl or tert-butyl, preferably methyl; and C 4 -C 8 cycloalkyl may be cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl or cyclooctyl, preferably cyclohexyl or cyclopentyl.

A process for the preparation of compounds of formula (Ia): compounds of formula (Ia)

R ₂ and R ₃ are as defined above; and

- X represents a chlorine atom or a bromine atom, by reacting with compounds of formula (IIIa) as defined above.

Suitably, the process is carried out by maintaining the compound (IIa) and the compound (IIIa) in a solvent such as dioxane at a temperature of about 80-120 ° C, preferably at the reflux temperature.

The X substituent of the compounds of formula (IIa) used as starting materials for this process is predominantly chlorine. The compounds of formula (Ha), together with a suitable process for their preparation, are disclosed in U.S. Patent No. 269,574. European Patent Publication.

(Ia) Compounds of formula I can also be prepared by reacting a compound which (IVa) wherein R ', R ₂ and R ₃ are as defined above with tetrabutylammonium fluoride trihydrate reacted.

This reaction can be performed by stirring in an organic solvent such as tetrahydrofuran at room temperature. Compounds of formula IVa can be prepared by reacting compounds of formula Ha with 1,3-dichloro-1,3,3,3-tetrazopropyldisiloxane in an alkaline solvent such as pyridine.

99

18 and then reacting the compounds thus obtained, wherein X, R ₂ and R _{3 in} formula (Va) are as defined above, with compounds of formula (IIIa).

The following 1-8. In the examples, all temperature data are given in 'C' without correction.

Example 1: 6- [(trans) -4-hydroxycyclohexyl] -2 ^{1 2} - (O-methyl) adenosine Preparation of 6- [(trans) -4-hydroxycyclohexyl] -9-purinyl-2 '- (0-methyl) -3', 5'-O- (1,1,3,3-tetraizopropyldisyloxy-1,3-diyl) -D-ribose, 10 g of tetrabutylammonium fluoride trihydrate and A mixture of tetrahydrofuran (20 ml) was stirred at room temperature for 15 minutes. The mixture was then evaporated to dryness and the residue was eluted on silica gel with ethyl acetate / methanol (85:15 v / v). The purified product was crystallized from a mixture of methanol and ethyl acetate. The title compound crystallizes from 1 equivalent of methanol, m.p. 121-124 ° C.

[α] D = -50.2 ° (c = 1; dimethylformamide).

The compound may also be recrystallized from ethanol and the compound will crystallize with 1 equivalent of water when left to stand in moist air. Melting point: 114-117 ° C.

[α] D = -46.9 '(c = 1, dimethylformamide).

6 - [(trans) -4-hydroxycyclohexyl] -9-purinyl-2 '- (0-methyl) -3', 5'-O- (1,1,3,3) used as starting material for the above procedure. The preparation of tetraizopropyldisyloxy-1,3-diyl) -D-ribose is described below.

g of 6-chloro-9-puridinyl-2 '- (O-methyl) -3', 5'-O- (1,3,3,3-tetrazopropyl-disyloxy-1,3-diyl) -D-ribose, 1.32g transII t • · ··· · 4 ···

- A mixture of 19 -4-hydroxycyclohexylamine, 1.4 ml of N-ethyldiisopropylamine and 80 ml of dioxane was stirred for 6 hours in a 105 ° C oil bath. The mixture was cooled to ambient temperature, filtered, and the filtrate evaporated to dryness. The resulting mixture was purified by elution on silica gel. Elution was carried out first with a 7: 3 by volume mixture of hexane and ethyl acetate and then with a 95: 5 by volume mixture of ethyl acetate and methanol. The resulting oil-like compound _Rf in ethyl acetate: methanol 95: 5 as determined by volume mixture of 0.3.

Example 2: Preparation of 6 - [(cis) -4-hydroxycyclohexyl] -2 '- (0-methyl) adenosine

The title compound was prepared as described in Example 1, but using cis -4-hydroxycyclohexylamine instead of trans-4-hydroxycyclohexylamine.

The compound thus crystallized with 1/2 equivalent of ethyl acetate, m.p. 110-111 ° C. [.alpha.] D @ 20 = -48.6 DEG (c = 1, dimethylformamide).

Example 3: Preparation of 6 - [(1 S, trans) -2-hydroxycyclopentyl] -2 (O-methyl) adenosine

The title compound was prepared as described in Example 1, but using (1S, trans) -2-hydroxycyclopentylamine instead of trans-4-hydroxycyclohexylamine. The foam-like title compound _Rf of ethyl acetate and ethanol as determined 75/25 mixture of 0, 44, [a] p ° value is -25.5 ° C (c = 1, dimethylformamide).

Example 4: 6- [(IR, trans) -2-hydroxycyclopentyl] -2 ¹ - (0-me-

• ·

- Preparation of 20 til) adenosine

The title compound was prepared as described in Example 1, but using trans (4R, trans) -2-hydroxycyclopentylamine instead of trans-4-hydroxycyclohexylamine. The title compound thus obtained has a melting point of 164-166 ° C.

_Rf in ethyl acetate: ethanol 75:25 as determined by volume mixture of 0.44 [a] _D value is ² ° -80.2 ° (c = 1, dimethylformamide diir.etil).

Example 5: Preparation of 5 - [(1 S, trans) -2-hydroxycyclohexyl] -2 (0-methyl) adenosine

The title compound was prepared as described in Example 1, but using (1S, trans) -2-hydroxycyclohexylamine instead of trans-4-hydroxycyclohexylamine. The foam-like title compound _Rf in ethyl acetate: ethanol 75:25 as determined by volume mixture of 0.42; [α] ²⁰ D = -26.2 ° (c = 1; dimethylformamide).

Example 6: Preparation of 6 - [(trans) -4-hydroxycyclohexyl] -2 '- (O-ethyl) adenosine

The title compound was prepared as described in Example 5. The compound 0.4 hydrate had a melting point of 98 ° C (liquefying) and a [α] ² ° of -58 ° (c = 1; dimethylformamide).

Example 7: Preparation of 6 - [(1 S, trans) -2-hydroxycyclohexyl] -2 ^1- (O-ethyl) adenosine

The title compound was prepared as described in Example 5.

Example 8: 6- [(R, trans) -2-hydroxycyclohexyl] -2 '- (O-me · · · · · · · · · · · ·

- Preparation of 21 til) adenosine

The title compound was prepared as described in Example 1, but using (1R, trans) -2-hydroxycyclohexylamine instead of trans-4-hydroxycyclohexylamine. The foam-like title compound _Rf in ethyl acetate: ethanol 75:25 as determined by volume mixture of 0.42; and [α] D -69.5 ° (c = 1, dimethylformamide).

The compounds of the present invention have valuable pharmacological properties and can therefore be used as medicaments, not only in the form of the formula I, but also in the form of their hydrates or addition products with organic solvents such as methanol, ethanol or ethyl acetate. Of particular interest to the compounds of formula (Ia) is the compound of Example 1, which is set forth below in Example 1. compound.

As mentioned above, the compounds of formula Ia are valuable anesthetics. These compounds of the invention may also be of interest because of their other effects, including the antihypertensive activity, as evidenced by the results of the following studies.

Binding to Al and A2 adenosine receptors in membranes derived from rat cortex or porcine cortex or corpus striatum R.F. BRUNS, G.H. LU and T.A. PUGSLEY method [MOLEC. PHARMACOL., 29, 331-346 (1986)]. Further testing of the activity of the compounds of formula I is carried out.

- performed in 22 isolated and flow-through rat kidneys by measuring the following parameters:

- renin formation,

- kidney hemodynamics (blood pressure measurements);

inhibition of the release of norepinephrine from nerve endings following electrical stimulation of the nerves in the kidneys [measurement Ρ. M. VANHOUTTE, D. BROWNING, E. COEN, T.J. VERBEUREN, L. ZONNEKEYEN and M. G. COLLIS, HYPERTENSION 4, 251-256 (1982)]; and

- changes in blood pressure, blood flow, blood flow, blood flow, and renal enzyme activity in normal and spontaneously hypertensive rats, who are awake, have lost sodium chloride in their blood, and are then supplemented with sodium chloride; catheters implanted by intravenous administration of the compounds of the invention as infusions or as large pills following the method of JFM SMITS and JM BRODY (Am. J. Physiol. 247, R1003-R1008 (1984)).

From the results of the studies it can be concluded that the inhibition of renin production, noradrenaline release from nerve endings and direct vasodilation play a role in the antihypertensive effect of the compounds of formula (Ia). In contrast to the decrease in blood pressure, urine production and electrolyte excretion remain unchanged. It follows that the compounds of formula (Ia) are not only reduced in hypertension.

- 23 but can also be used to widen coronary vessels. In addition, said compounds protect the endothelium of blood vessels by inhibiting platelet adhesion and activating white blood cells. Compounds of formula (Ia) also lower blood lipid concentrations.

In the case of the above indications, the compound of formula (Ia) is preferably the compound of example 1 above. The compounds of formula (Ia) are primarily intended for cardiac protection and analgesia.

Furthermore, can be used for treating irregular heart rhythms in patients due to Formula (Ia) Compounds of formula adenosine A ₁ receptor agonist activity of goods, which - as is to be described selective such later - unlike their activity against receptor ^- A ₂

a) It is evident from the test and, for example, its ability to extend the passage through the atrial ventricular (A-V) node of the heart, as will be described later

b) shown by test results. Thus, these compounds restore sinus rhythm in supaventricular tachycardias, particularly in seizures, in reducing seizure in tachycardial atrial tremor, and in restoring normal heart rate in t-adrenergic pacing.

The effects of the compounds of formula (Ia) are similar to those of preconditioning, that is, the effect of a conditioning treatment which is a short-term local anemia condition.

- With 24 inducers, it makes the heart resistant to a subsequent infarction that threatens anemia. The use of these compounds can therefore protect the heart, with or without complementary therapies that result in the resolution of blood clots, against myocardial infarction or many anemia caused by anemia, which endangers patients who have, for example, coronary artery bypass grafting, vascular surgery. surgery, heart transplantation, or other non-cardiac surgery.

The compounds of formula (Ia) are particularly useful in the treatment of persons who are predisposed to myocardial infarction, for example, or who have already undergone a myocardial infarction.

The compounds of formula (Ia) further reduce plasma insulin without affecting glucose tolerance. These compounds also potentiate the effect of insulin in increasing adipose glucose uptake and suppressing fat breakdown, which results in a reduction in the concentration of free fatty acids in the plasma as shown in the test d) below. However, a decrease in free fatty acids in the plasma results in a decrease in the triglyceride concentration and consequently in an increase in HDL cholesterol.

By virtue of their insulin-saving and / or lowering concentration of free fatty acids, the compounds of formula (Ia) can be used for the first time.

- treating diabetes mellitus and type 2 diabetes, non-insulin dependent diabetes mellitus.

Since compounds of formula (Ia) lower plasma triglyceride and free fatty acid levels (as shown in test e) below and increase plasma HDL cholesterol levels, lipid dysfunction and for the treatment of conditions associated with elevated plasma free fatty acids and triglycerides, and in cases where an increase in HDL cholesterol is desired.

The compounds of formula Ia have good metabolic stability.

As a result of the present invention, it is also possible to use the compounds of formula (Ia) in the manufacture of medicaments for the treatment of analgesia or analgesia.

The present invention also results in the availability of analgesic compounds of formula (Ia) as defined above.

As a result of the present invention, analgesic compositions containing a compound of formula (Ia) as hereinbefore defined and a pharmaceutically acceptable carrier or diluent may be prepared. The use of compounds of formula Ia according to the indications already described for compounds of formula I is contemplated.

• · ·

The following pharmacological tests illustrate the activity of the compounds of formula (Ia) as previously described.

a) Affinity for adenosine receptors

Membranes from porcine corpus striatum were prepared as described by H. Bruns et al. Bruns et al. (1986) Molecular Pharmacology 29: 331-344. ³ H-NECA, a non-selective adenosine receptor agonist, is used to label both A ₁ and A ₂ receptors. IC ₅₀ values are determined from the displacement curves of the non-linear weighted curve fitted to the Langmuir equation and the calculated pK _D values.

Compounds of formula (Ia) exhibit affinity for the A 1 receptor, for example, when present at a concentration of 1 to 500 nM.

b) Arrhythmias

Activation of the adenosine A ₁ receptor reduces, for example, the incidence of supraventricular tachycardia, tachycardial ventricular tremor, and other arrhythmias in seizures by slowing down the flow of PR at the node of the atrial ventricular (AV) node. presents. The assay involves recording ECG changes in conscious Rhesus monkeys. The compounds of formula (Ia) are administered orally at a dose of about 0.1 to 1000 mg / kg or intravenously at a dose of 0.03 to 30 mg / kg, according to the method of C. Clarke et al., The Pharmaceutical Journal, 244, 595- 597 (1990)]. For example, in this assay, oral administration of doses of 0.1, 0.3 and 0.6 mg / kg orally is shown in FIG. compound increases the PR interval of the surface electrocardiogram (ECG), indicating slower flow through the AV node. As mentioned above, this effect prevents the tachycardia from recurring at the AV node and reduces the heart rate in the case of tachycardial atrial tremor.

c) Pre-conditioning protection against infarction

Preconditioning (5 minutes of anemia followed by 10 minutes of regeneration) greatly increases cardiac resistance to the subsequent 30 minutes of anemia and subsequent infarction for 3 hours. The test method is known from two articles [G. S. Liu et al., Circulation, 84, (1), 350-356 (1991) and J.D.

Thornton et al. (1992) Circulation 85: 659-665. The compounds of formula (Ia) are administered intravenously at a dose of about 0.01 to 10 mg / kg. In this study, no. Rabbits receiving 100 mg / kg of Compound II are almost as protected as rabbits preconditioned for myocardial infarction by occluding the coronary artery for 30 minutes.

d) Glucose transport and fat digestion in rat fat cells

i) Collagenase digestion isolates fat cells from adipose fat pads of rats fed a normal diet. Cells with a final concentration of 2% v / v with adenosine deamylase (1 U / ml), the test compound, e.g. compound, and other additives recommended for glucose transport measurement, are preincubated at 37 ° C.

After ³ min, [3- ³ H] glucose (final concentration: 50 μΜ; 0.5 gCi / mL) was added and incubation continued for another 60 min. The incorporation of radioactive [3- ³ H] glucose into lipids in cells, which is a measure of glucose transport, was determined by extracting 0.5 ml of cell suspension with 5 ml of toluene-based scintillant and counting by liquid scintillation. The water-soluble metabolites, together with the remaining [3- ³ H] glucose, remain in the aqueous phase and thus do not interfere with the measurement.

Fat degradation is measured by some conventional method. In one method, cells are incubated in the absence of isoproterenol and with 1 μΜ of isoproterenol. After centrifugation through dinonyl phthalate, the supernatant is separated from the cells and the degree of lipid degradation is determined by enzymatic determination of the amount of glycerol released into the supernatant.

Compounds of formula Ia are active in the concentration range of 0.1 to 1000 nM.

No. 1 Compound II inhibits lipolysis in rat adipose cells at concentrations of 0.1-100 nM.

In the presence of adenosine deaminase (1 U / ml) Compound does not have a significant effect on radioactive [3- ³ H] 29

-glucose uptake into lipids in cells in the absence of insulin, and the effect is negligible even when insulin is present at maximal stimulatory concentration (8 nM) but significantly increases the concentration-dependent uptake of the radioactive compound when insulin is present. , At a concentration of 1 to 50 nM. These observations suggest that no. compound increases the sensitivity of glucose transport to insulin. In the event that no. is present at near maximal effective concentrations, the EC ₅₀ for insulin stimulation of [3- ^J H] glucose by lipids is reduced to one-half to one-third. In the presence of adenosine deaminase enzyme and 1 μΜ of isoproterenol, 10-50 nM no. compound increases both the sensitivity and the degree of insulin response; the EC ₅₀ for insulin can be reduced by up to a fifth, and the stimulant effect of insulin at maximum concentrations is greatly increased.

ii) Development of lipid and glucose levels in normal 18-hour fasted rats

Rats of 2-3 months, about 250 g, are kept for 7 days in a room set at 22 ° C, where light and dark periods follow each other in 12/12 hour cycles.

The rats may consume any amount of purine rat food and water as desired. After 18 hours of fasting, rats (5 rats / group) were injected with 1.0 mL / 100 g body weight of test compounds in carboxymethylcellulose 0.5 x · · ·

- 30% by weight of the mixture. After 3 hours after administration of the test compounds, the rats were anesthetized with carbon dioxide and blood was drawn from them by cardiac aspiration. Blood sera were collected to determine concentrations of glucose, free fatty acid, and β-hydroxybutyrate. Free fatty acids were assayed by an acyl-COA-peroxidase calorimetric enzyme assay, glucose by the glucose oxidase method (YSI Model 27, Yellow Spring, Ohio), and β-hydroxybutyrate by a β-hydroxybutyrate dehydrogenase enzyme assay (Sigma Kit 310- A - St. Louis, Mo.). The compounds of formula (Ia) are effective at doses of from about 1 to about 5000 pg / kg.

2 hours after the administration, it can be stated that no. When administered at doses of 5-100 μg / kg, Compound II reduces the free fatty acid content as a major result of its action on fat cells. As a result, β-hydroxybutyrate and blood glucose are decreased in a dose-dependent manner.

iii) Effects on non-insulin dependent diabetes mellitus (NIDD) rats

During the NIDD screening test, rats weighing 200 g each are fed a high-fat diet, which can be consumed as desired. 41 mg of streptozotocin / kg body weight were injected into the tail vein of rats fed in this manner. After 1 week, rats with dietary blood glucose levels greater than 200 mg / dl and those having a blood glucose level of 40-80 mg / dl after 3 hours of fasting overnight are considered diabetic. Four days later • · ·

- 31 animals with blood glucose levels greater than 180 mg / dl are screened. Blood glucose levels were determined with a YSI Glucose Analyzer. Performing a chronic screening assay is described below.

At 9 am on the first day, the rats are deprived of food. Initial glucose was determined in blood samples taken from the caudal tip, followed by oral administration of the test compound to 9 rats in the control group, and to test compounds in the 9-9 rats in the other groups. After six hours, the blood glucose is measured and immediately afterwards the rats are fed again.

The rats referred to in the previous paragraph are administered once daily for a period of 11 consecutive days with vehicle or drug. Blood glucose levels were determined at 0 hours, and at the end of fasting for 6 hours after dosing on days 4, 8 and 11. Compounds of formula (Ia) administered daily at a dose of 100 μg / kg / day for 11 days, e.g. Compound - significantly reduces the amount of free fatty acids in the plasma resulting in a significant decrease in blood glucose.

e) Dyslipidemias due to increased levels of triglycerides in the serum (disorders due to increased fat in the blood)

Some studies have shown that there is a positive correlation between serum triglyceride levels (and consequently reduced HDL cholesterol) and the risk of developing coronary heart disease (CHD) [Grundy:

«·

- 32 Cholesterol and Atherosclerosis: Diagnosis and Treatment, Lippincott, Philadelphia, 1990]. The Helsinki Heart Study clearly showed how reducing high triglyceride levels could be one of the possible ways to reduce the risk of CHD. In this study, the rate of major coronary artery disease reduction after treatment with gemfibrozil was the highest in type IIB hyperlipidemic patients with high levels of both LDL cholesterol and total serum triglycerides and generally low HDL cholesterol. . The compounds of formula Ia are about 0.03 to 30 mg / kg, e.g. 0.1 to 30 mg / kg, when administered intravenously to Rhesus monkeys, and 0.1 to 100 mg / kg, e.g. , Effective at doses of 1 to 10 mg / kg. No. 1 Compounds of formula IV, when administered intravenously in rhesus monkeys at doses ranging from 0.03 to 0.6 mg / kg and orally from 0.1 to 1.2 mg / kg, decrease the concentration of free fatty acids and triglycerides in the serum in a dose-dependent and sustained manner. No. 1 For example, compound 0.60 mg / kg, administered orally, reduced the free fatty acid content by about 60% and the triglyceride content by 40% after 300 minutes compared to the control.

Compounds of formula (Ia) also exhibit activity in anesthetized rats with moderate arterial blood pressure, low heart rate, and peripheral vasodilation.

The experiments were performed intraperitoneally on 120 W / kg male Wistar rats anesthetized with pentotal at a dose of 120 mg / kg, according to the method described by Salzman et al. Cardiovasc. Pharmacol. 12: 451-460 (1988). Catheters are inserted into the right cervical vein, the right femoral vein, the left ventricle (introduced through the right carotid artery), the left thigh artery, and the aorta (introduced through the right thigh artery). The following parameters are measured or calculated: systolic, diastolic, and mean arterial blood pressure (Hgmm; left thigh artery blood pressure, Statham P 23 Gb pressure transducer), heart rate (Hgmm), heart rate (heart rate / min), growth rate of left ventricular pressure (dP / dt _max , mmHg / s; Statham P 23 Gb pressure transducer), cardiac percolume [ml / (min-100 g body weight), thermodilution method, right cervical vein and aorta], complete peripheral resistance (dynes s ^cm-5/100 g body weight) and surface ECG. Arterial blood pressure, left ventricular pressure, dP / dt _max , heart rate and electrocardiogram are recorded continuously with a Schwarzer polygraph (i.e., a device for simultaneous recording of blood pressure, artery and respiration). Parameters were measured 30, 20, 10, and 2 minutes before test compounds application, and 1, 5, 10, and 15 minutes after injection of test compound into the right femoral vein. Compounds of formula (Ia) are injected at a dose of about 0.001 to about 10 mg / kg body weight. No. 1 Compound II was tested at increasing doses of 0.003 mg / kg, 0.010 mg / kg and 0.03 mg / kg, with three experimental animals34 per dose. No. 1 Compound II reduces blood pressure (ED ₅₀ = 49 µg / kg by intravenous administration) and heart rate, and increases systemic vascular conductance.

Dosages of the compounds of formula (Ia) used in accordance with the claimed indication will vary in the usual manner depending on the severity of the disorders, the weight of the patient and the relative efficacy of the compound employed. However, as a general guideline, suitable unit doses within the 0.1 to 1000 mg range include, for example, 0.1 to 10 mg, 0.5 to 200 mg, 0.5 to 100 mg. or in the range of 0.5 to 10 mg. For example, unit doses of 0.1 mg, 0.5 mg, 1 mg, 2 mg, 3 mg, 4 mg and 5 mg may be administered more than once a day - for example 2, 3, 4, 5 or 6 times, but once or twice daily administration is preferred. Thus, a total daily dose of a 70 kg mammal, such as a human, is, for example, about 0.1 to 1000 mg for oral administration and 0.03 to 300 mg for intravenous administration; that is, the dose is in the range of about 0.001 to 20 mg / (kg-day), e.g., 0, 007 to 3 mg / (kg-day), 0, 007 to 1.4 mg / (kg-day), 0.007. - in the range of 0.14 mg (kg-day) or 0.01-0.5 mg / (kg-day) - so for example 0.01, 0.02, 0, 04, 0.05, 0, 06, 0, 08, 0, 1, or 0.2 mg / (kg-day) - and treatment with such doses may take several weeks or months. In the case of an adult weighing 70 kg, no. The compound of the present invention is preferably administered in a dosage of 0.1 to 10 mg / day for all indications of the invention. In the treatment of non-insulin dependent diabetes mellitus and hypertriglyceridemia which is preferable to treat, the oral dosage is 0.2 to 2 mg / day in humans and 0.2 to 10 mg / day for oral heart failure and other cardiac disorders, preferably 0, 5-5 mg / day, and 0.25-5 mg / day for intravenous treatment of arrhythmias such as tachycardia atrial tremor.

The compounds of formula (Ia) may be formulated for use in any convenient manner. Depending on the disorder requiring treatment, what method is best used for shaping. The formulations may be presented in unit dosage form or in such a manner that the patient may self-administer a single dose. Compounds of formula Ia, as described above for use as analgesics, are preferably administered orally, rectally, topically, parenterally, by intravenous or intramuscular injection.

For oral administration, unit dosage forms may be tablets and capsules containing 0.05 to 20 mg of a compound of formula (Ia).

Suitable amounts of the compounds of the present invention, or hydrates thereof, or addition products thereof with organic solvents, are suitably in the range of about 0.5 to 20% by weight, preferably about 1 to 10% by weight, for example 2 to 5% by weight.

Compounds of formula (Ia) wherein R ' _T is hydroxy (C 4 -C 8 cycloalkyl) and R ₃ is alkyl are found to be metabolites when R ₃ is a C 4 -C 8 cycloalkyl group and R ₃ is alkyl. The compounds of formula (I) are administered to warm-blooded animals such as rats, dogs and humans.

For example, using compound (M) gives hydroxycyclohexyl-2 '- (O-methyl) -adenosine.

Object of the present invention was also possible to apply a method, wherein ^{N-cyclohexyl-phenyl} 2 '- introducing (0-methyl) adenosine organism in warm-blooded animals that ^{N6-hydroxycyclohexyl-2'} - (O-methyl) - adenosine.

The use of the present invention also makes it possible to prepare N ⁶ - (hydroxycycloalkyl) -2 '- (O-alkyl) adenosine in pure form, for example, in a purity greater than 95% by weight. Exemplary compounds described herein meet this requirement.

It is also possible to prepare N ⁶ - (hydroxycyclohexyl) -2 '- (O-alkyl) -adenosine free of cyclohexyl-2' - (O-methyl) -adenosine using the invention.

The present invention also provides a process for the preparation of 2 '- (O-alkyl) -adenosine derivatives, which comprises treating adenosine with the appropriate alkyl sulfate in the presence of a phase transfer catalyst.

As a 2'- (O-alkyl) -adenosine derivative, it is desirable to prepare one of the above-mentioned compounds.

The present invention also relates to a process for the preparation of compounds of formula I wherein the compounds of formula VI are in a non-polar solvent (R ₃ ) ₂ SO ₄

After treatment with a 37 basic solution in the presence of tetrabutylammonium hydrogen sulfate, the resulting product is purified by recrystallization.

If necessary, reactive groups may be temporarily protected.

Suitable alkyl sulfates are di (C 1 -C 4) alkyl sulfates.

The phase transfer catalyst is preferably tetrabutylammonium hydrogen sulfate.

The reaction may conveniently be carried out in a non-polar solvent, such as one of the solvents described below.

A separate group of compounds of formula I comprises those compounds wherein R 1 is C 3 -C 8 cycloalkyl, R ₂ is hydrogen or C 1 -C 4 alkyl, and R ₃ is C 1 -C 4 alkyl.

To date, the compounds of formula (I) have been prepared, usually in six steps, as follows: preparation of 2 ', 3', 5'-triacetylinosine; Preparation of 6-chloro-9-pD-ribofuranosyl-9H-purine by chlorination and hydrolysis; protecting the 3'-O- and 5'-O-positions with tetrazopropyldisiloxane groups (TIPDS-CL ₂ ); 2'-O-alkylation and purification by silica gel chromatography; removing the protecting groups from the 3'-0 and 5'-positions; and reacting the resultant product with a compound of formula R _X NH ₂ followed by recovery of the compound of formula I by recrystallization.

We have found that the compound of the formula I •

38 runs can be prepared in high yield and high purity without disiloxane protection and deprotection of the 3'-O and 5'-O positions, or without chromatography on silica gel. It has been found that the inosine-2 ', 3', 5'-triacetate starting material used to carry out the processes used so far can be advantageously substituted by the lipophilic inosine-2 ', 35'-tripropionate. This process doubles the productivity and avoids the use of unwanted pyridine solvents.

According to the invention, the compounds of formula (I) are prepared under conditions sufficient to phase transfer catalysts in the reaction according to Scheme 1, wherein R _lz R ₂ and R ₃ are as defined above.

The compounds of formula (I) may be prepared by reacting an alkaline aqueous solution of the compounds of formula (VI) with di (C 1-4 alkyl) sulfate in a water immiscible organic solvent in the presence of tetrabutylammonium hydrogen sulfate. An alkaline metal hydroxide such as sodium hydroxide or potassium hydroxide may be used to prepare the alkaline aqueous solution. The solvent may be any water immiscible or substantially immiscible solvent in which the compound of formula (I) is soluble; such as methylene chloride or tertiary amyl alcohol. The reaction is preferably carried out at a temperature of about -20 ° C to about 50 ° C, in particular room temperature. Reaction time is not a critical parameter, but it is preferred that the reaction time be about

5-10 hour intervals and more preferably about ·

• · «· • <

• · »·

- 39 is in the 7-8 hour interval. The crude products of formula (I) are isolated by evaporation of the reaction mixture, and the residue is stirred with a 1: 1 mixture of water and an inert solvent, preferably toluene, at ambient temperature for 5-7 hours, and the precipitated material is filtered off and dried. . The pure compounds can be obtained by fractional crystallization, which is conveniently carried out by:

1. recrystallizing the crude product from an inert solvent such as toluene by dissolving the crude product at 80 ° C in the solvent, maintaining the solution at about 55 ° C for about 1 hour, cooling to ambient temperature and crystals of the pure compound and filtering off the resulting crystals;

2. repeating the recrystallization step of step 1, but maintaining the solution at 65 ° C before cooling to ambient temperature, and

3. Recrystallization from 100% ethyl alcohol by dissolving the product under reflux, diluting with water, inoculating, and drying the crystals formed after filtration.

Many of the compounds of formula (VI) are known and are described in the literature, e.g. and 4,985,409. U.S. Pat. The compounds of formula (VI), as already mentioned, can be conveniently obtained by the process of Scheme 2, which is conveniently employed. Scheme 2 is a replicate R _; and R ₂ are as defined above.

Thus, compounds of formula (VI) may be prepared by acylation of the inosine derivatives of formula (VII) with propionic anhydride to give the 2'-O-, 3'-O- and 5'-O-protected compounds of formula (VIII). halogenation with thionyl chloride and simultaneous amination and hydrolysis of the resulting intermediates of formula IX to give compounds of formula IX. The acylation of inosine is conveniently carried out in a mixture containing toluene, tributylamine and 4-dimethylaminopyridine at 100-110 ° C for 4-5 hours. The compounds of formula VIII are isolated by precipitation with heptane. The compounds of formula (VIII) are suitably halogenated in a mixture of toluene and N, N-dimethylformamide at 60-70 ° C for 3 to 4 hours, and the solution of the compounds of formula (IX) is then washed with aqueous and brine. hereafter. The simultaneous amination and hydrolysis of intermediates of formula (IX) is conveniently accomplished by the addition of an amine of formula R ₁ NH ₂ and the reaction mixture maintained at a temperature of 100 ° C to 110 ° C for 15 to 20 hours. The compounds of formula (VI) are isolated by filtration at ambient temperature and purified by recrystallization.

The following examples illustrate the process of the invention.

• · ·

- 41 Production Example 1

Preparation of N ⁶ -cyclohexyl-2 ^1- (O-methyl) -adenosine

Step A: Preparation of inosine 2 ', 3', 5'-tripropionate

A mixture of 271.2 g of inosine, 966 ml of tributylamine, 3.30 g of 4- (dimethylamino) pyridine and 600 ml of toluene is heated to an internal temperature of 104-105 ° C and 453 ml of propionic anhydride are added over 35 minutes. at a rate such that the internal temperature remains between 104 ° C and 105 ° C. The mixture was stirred at this temperature for an additional 4 hours, then cooled to 5-10 ° C in an ice bath, and heptane (1000 mL) was added. The resulting suspension is stirred at ambient temperature, 20-22 ° C, for 30 minutes and then filtered, for example, in a Buchner funnel. The solid was washed with a total of 450 ml of heptane in three equal portions of 150 ml and dried overnight (14 hours) at 45-50 ° C under 3333 Pa. 425.9 g (96.5%) of inosine 2 ', 3', 5'-tripropionate are thus obtained in the form of a white solid.

Step B: Preparation of N ⁶ -cyclohexyladenosine

A mixture of 270.6 g of inosine 2 ', 3', 5'-tripropionate, 240 ml of N, N-dimethylformamide and 600 ml of toluene was heated at 65 ° C for 1 hour and then added at a rate of 67.84 g. of thionyl chloride to keep the internal temperature between 62 ° C and 65 ° C. The reaction mixture was stirred at this temperature for a further 2.5 hours and then cooled to 10 ° C in an ice bath. Then add 600 ml of water cooled in a pre-ice bath at 10-15 ° C at a rate such that the temperature of the mixture is below 20 ° C. The organic layer was separated and washed with a total of 800 mL of 10% w / w aqueous sodium chloride in 200 mL portions. The crude organic layer containing 6-chloro-9- [2,3,5-tri- (O-propionyl) -3-ribofuranosyl] -9H-purine was added with stirring over 2 hours to a temperature of 105 ° C (620 ml). cyclohexylamine, at a rate such that the internal temperature remains at 105 ° C. The resulting mixture was stirred at this temperature for an additional 17 hours, then cooled to ambient temperature - 25 ° C with vigorous stirring for about 2 hours - and then filtered, for example, on a Buchner funnel. The solid containing N ⁶ -cyclohexyladenosine and cyclohexylamine hydrochloride was washed with a total of 460 mL of four equal volumes of 115 mL of toluene and transferred wet into a 5 L flask mixed with a mechanical stirrer. Aqueous saturated sodium bicarbonate solution (2 L) and ethyl acetate (2.5 L) were added and the resulting mixture was stirred for about 10-15 minutes until the total solids dissolved. The organic layer was separated and the aqueous layer was washed with a total of 1.5 L, first 1 L, then 500 mL, ethyl acetate.

_f o 44

The organic layers were combined and evaporated at 40 DEG C. under 10-10 bar until ethyl acetate (about 3 liters) was removed. Heptane (500 mL) was added to the residue and the resulting mixture was stirred for 30 min. The solids were separated on a Büchner funnel, and a total of 300 mL was used in three equal 100 mL portions of hep • · ·

- 43 tannas wash. The solids were then 3-10 ³ -3,5-10 ³ mm Hg for about 3 hours at 45-50 ° C. This gave 148 g of crude N ⁶ -cyclohexyladenosine as a white solid, which was transferred to a 1 liter round bottom flask equipped with a mechanical stirrer and 175 ml of 95% ethanol was added. The resulting slurry was stirred for 15-20 minutes and then tert-butyl ethyl ether (175 mL) was added. The suspension was stirred for 5 minutes, then cooled in an ice bath and stirred for an additional 15 minutes. The suspension was filtered through a Buchner funnel and the solid was washed with a total of 50 ml of two equal portions of 25 ml of tert-butyl methyl ether.

The filtered solid was dried at 45-50 ° C under a pressure of 3 x 10 ³ -3.5 x 10 ³ Pa for 14 hours. This gave N ⁶ -cyclohexyladenosine (130 g, 60.0% yield) as a white solid, m.p. 185-187 ° C.

Step C: Preparation of N ⁶ -cyclohexyl-2 '- (O-methyl) adenosine

A suspension of 94.33 g of N ⁶ -cyclohexyladenosine and 720 g of 5% w / w aqueous sodium hydroxide solution was stirred at ambient temperature - 24-25 ° C (about 5 minutes) until the solid the total amount does not dissolve. 850 ml of dichloromethane and 5.5 g of tetrabutylammonium bisulfate are added via a funnel, followed by 61.3 g of dimethyl sulfate over a period of 5 to 10 minutes at an internal temperature of 24-25 °. C stay. The addition funnel was washed with an additional 50 mL of dichloromethane and this dichloromethane was added to the reaction mixture. The biphasic mixture is maintained at an internal temperature of 24-25 ° C.

- 44 stirred for 7.5 hours, then the organic layer was separated and 40 ° C until concentrated to 2.7 to 10 ⁴ 2.9 to 10 Pa ⁴ Pa until further quantities solvent stops coming out from the mixture. The residue is dissolved in 200 ml of toluene and the resulting solution is evaporated at 45-50 [deg.] C. under 4 kPa until no more solvent is distilled off.

A mixture of the above crude product and 2470 mL of toluene was stirred at ambient temperature for 10 minutes and then 2470 mL of water was added over 22 minutes. The resulting slurry is stirred at ambient temperature for an additional 6 hours and the solid collected by filtration, for example by baking on a Buchner funnel. The solid was washed with toluene (114 mL) followed by a total of 285 mL of three equal portions of water (95 mL) and dried at 48-50 ° C overnight (14 h). This gave 53.0 g of a white solid which was suspended in 397 ml of toluene and stirred at 80 ° C. The resulting clear solution was cooled to 56 ° C over 45 minutes and seeded with 10 mg of pure product. The resulting mixture was stirred at 55-56 ° C for 45 minutes and then cooled to ambient temperature over 1 hour. The mixture is stirred at this temperature for an additional 1 hour and the solid is collected by filtration, for example by Buchner funneling. The solid was washed with a total of 75 mL of three equal portions of 25 mL of toluene. This gives 60 g of a white solid which is suspended in 159 ml of toluene and the resulting suspension is heated to 80 ° C. The inoculation described above is then carried out at 65-66 ° C.

The mixture was cooled to ambient temperature over 1 hour and stirred at this temperature for an additional 1 hour. The solid was filtered off and washed with a total of 42 ml of three equal portions of 14 ml of toluene and dried at 48-50 ° C overnight (14 hours). 57.7 g of a white solid are obtained. To this solid was added 122 mL of 100% ethanol and the resulting mixture was refluxed until a clear solution was formed. To the solution was added 288 ml of water preheated to 55 ° C over 25 minutes. The resulting mixture was cooled to 55 ° C and seeded with 20 mg of pure product. The resulting mixture was cooled to ambient temperature over 1 hour and stirred at this temperature overnight (16 hours). The solid was collected by digestion on a Buchner funnel and washed with a 1: 2.36 (v / v) mixture of 100% ethanol / water. A total of 57 ml of the mixture was washed in three equal 19 ml portions. The solid was dried at ambient temperature at 3866 Pa. 62.2 g (42.5%) of the title compound are obtained as a white solid with 1.5 molecules of crystalline water, m.p. 88-91 ° C.

Production Example 2

N ⁶ -cyclopentyl-2 (O-methyl) -adenosine was prepared as described in Preparation Example 1 except that an equivalent amount of N ⁶ -cyclopentyl-adenosine was used.

Production Example 3

The procedure of Example 1 is repeated, except that in step C, tertiary amyl alcohol is used in place of dichloromethane and, after crystallization from the first mixture of ethanol and water, the product is recrystallized from ethanol / water. 32.1 g of N ⁶ -cyclohexyl-2 '- (0-methyl) -adenosine are thus obtained in a purity of more than 98% by weight, about 30%.

The process of the present invention is more economical in terms of time and cost than prior art processes. The process provides an appropriate method for the selective 2'-O-methylation of N ⁶ -cyclohexyladenosine under phase transfer conditions and the purification of the 2'-O-methyl derivative. This procedure eliminates the need for expensive operations involving the use of protecting groups as well as chromatography.

Claims (8)

Claims Use of compounds of formula (I) - in formula (I) - R ₃ is hydrogen, C 1-4 alkyl, allyl, metallyl, straight or branched C 3-7 alkynyl, C 3-8 cycloalkyl, hydroxy (C 4-8 cycloalkyl), including 9 and Phenyl, singly or twice independently substituted with a halogen atom, a C 1-4 alkyl group, a C 1-4 alkyl group, a C 1-4 alkoxy group or a trifluoromethyl group; phenyl (C 1 -C 4) alkyl, together with the halo, C 1 -C 4 -alkyl, C 1 -C 4 -alkoxy or trifluoromethyl, singly or independently of such substituents, on the phenyl ring doubly substituted phenyl (C1-C4) alkyl; C 1-4 alkyl substituted with at least one hydroxy group or at least two phenyl groups; a bicycloalkyl group such as endo or exo-bicyclo [2.2. l] heptyl -; naphthyl (C1-C4) alkyl, acenaphthylenyl (C1-C4) alkyl; or a group wherein, in the general formula (A) or (B), - Z is hydrogen, hydroxy or (C 1 -C 4) -alkoxy; - Q is hydrogen or hydroxy; - A represents a methylene group, an oxygen atom, a sulfur atom or a direct bond; - Y is a direct bond or - (CH ₂ ) _n - wherein n is 1, 2 or 3; and the dotted line in formula (A) represents an optional bond; and - R ₂ is hydrogen, (C 1 -C 4) -alkyl, amino, (C 3 -C 5) -cycloalkyl, or a halogen atom of 9 to 35 inclusive; and - R ₃ is C 1-4 alkyl as anesthetic agents for the manufacture of a medicament for the treatment of pain. 2. Adenosine derivatives having the general formula (Ia) - _Rf 'is a substituted C4-8 cycloalkyl, hydroxy; R ₂ is as defined in claim 1, and R ₃ is as defined in claim 1. 3. Use of a compound of formula (Ia) as defined in claim 2 for the preparation of a medicament for the treatment of pain, reduction of hypertension, dilation of coronary arteries, prevention of platelet adhesion and activation of white blood cells. - 49, to lower blood lipid levels, to treat congestive heart failure, to prevent myocardial infarction, to prevent sudden cardiac death, to treat kidney failure, to treat hypertriglyceridemia and to raise low HDL cholesterol, to prevent high levels of lipid dysfunction, of type I and type II. type 2 diabetes, to control abnormal heart rhythm and / or to prevent myocardial infarction. Pharmaceutical compositions comprising, in addition to pharmacologically acceptable carriers or diluents, a therapeutically effective amount of a compound of formula (Ia) as defined in claim 1. 5. A method for treating pain, reducing hypertension, dilating coronary artery, preventing platelet adhesion, lowering high blood lipids, treating congestive heart failure, preventing sudden cardiac death, treating low-grade cholesterol, hypertriglyceride to reduce the concentration of free fatty acids in blood, type I or type II. Type I diabetes, treating cardiac arrhythmias, preventing myocardial infarction, comprising administering to a patient in need thereof a compound of formula (Ia). • · - 50 6. A process for the preparation of 2 '- (O-alkyl) -adenosine derivatives comprising mixing adenosine with appropriate alkyl sulfates in the presence of a phase transfer catalyst. Compounds prepared by the process of claim 6. 8. A process for the production of N ⁶ - (hydroxycyclohexyl) -2- (0'-methyl) -adenosine, which comprises administering N ^b -cyclohexyl-2 (0-methyl) -adenosine to warm-blooded animals.