Classifications

• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
  • C07H19/00—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof
  • C07H19/02—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof sharing nitrogen
  • C07H19/04—Heterocyclic radicals containing only nitrogen atoms as ring hetero atom
  • C07H19/16—Purine radicals
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P25/00—Drugs for disorders of the nervous system
  • A61P25/04—Centrally acting analgesics, e.g. opioids

Definitions

• This invention relates to adenosine derivatives, and in particular, provides a new use of A1 receptor agonists.
• R 1 is hydrogen, (C ⁇ alkyl, allyl; methallyl; a straight-chain or branched
• Z is hydrogen, a hydroxy group or a (C 1 _ 4 )alkoxy group
• Q is hydrogen or hydroxy
• A is -CH 2 -, -0-, -S- or a direct bond
• Y is -(CH 2 ) n - or a direct bond, n is a whole number from 1 to 3
• R 2 is hydrogen, (C 1w ,)alkyl, amino, (C ⁇ cycloalkyl or halogen with an atomic number of 9 to 35 and R 3 is (C ⁇ )alkyl.
• R 1 is other than hydroxycycloalkyl.
• R. is hydroxy(C 4-8 )cycloalkyl.
• the compound of formula I is of 6-cyclohexyl-2'-0-methyl-adenosine, 5 hereinafter referred to as Compound M.
• Pig striatal membranes are prepared as previously described by H. Bruns et al. in Molecular Pharmacology 29 (1986) pages 331-344.
• IC 50 values are derived from the displacement curves by weighted non-linear least-square curve fitting to the Langmuir equation and pK D values calculated.
• CV 1808 2-Phenylaminoadenosine (Carbohydrates vol. 81 1974) ref. 91898 K)
• CGS 21680 2-[p-(2-Carboxyethyl)phenethylamino]-5'-N-ethyl carboxamido-adenosine (FASEB J, 1989, 3 (4) Refs. 4770/3)
• CPA proves to be a potent and highly selective displacer of binding to the A. receptor
• CV 1808 is a relatively weak but selective A 2 receptor ligand
• CGS 21680 shows high potency and selectivity for the A 2 receptor.
• Compound M in the form of its 1.5 hydrate shows good affinity and high selectivity for the A 1 receptor vis a vis the A2 receptor.
• the known compounds of the formula I are known as antihypertensive agents and coronary vasodilators.
• the compounds of formula I protect the vascular endothelium by inhibiting both thrombocyte aggregation and activation of leucocytes. They also lower the blood lipid levels. Further, compounds of formula I have a protective effect against diseases caused by hypertension such as congestive heart failure, myocardial infarction or sudden cardiac death and renal insufficiency (see Europ. Pat. Appl. EP-0-378 518 and EP-0-269 574).
• peripheral neuropathies such as diabetic neuropathy and of disorders associated with peripheral vascular diseases and/or disorders associated with neuronal degeneration, hypertriglyceridemia/low HDL cholesterol levels, lipid dysfunction, elevated free fatty acids or type I or type II diabetes including non-insulin dependent diabetes, arrhythmias in particular paroxysmal supraventricular tachycardia and tachycardiac atrial fibrillation and for protection against myocardial infarction.
• arrhythmias in particular paroxysmal supraventricular tachycardia and tachycardiac atrial fibrillation and for protection against myocardial infarction.
• the compounds of formula I are particularly interesting analgesics, for example for the treatment of pain, e.g. acute or chronic pain.
• the analgesic activity of the compounds of the formula I are indicated by their analgesic activity in standard animal tests, e.g. inflammatory and neuropathic models e.g. in reducing persistent inflammatory mechanical hyperanalgesia [(tests a) and b)] and persistent neuropathic thermal hyperalgesia [test c)] indicative of chronic neuropathic pain.
• standard animal tests e.g. inflammatory and neuropathic models e.g. in reducing persistent inflammatory mechanical hyperanalgesia [(tests a) and b)] and persistent neuropathic thermal hyperalgesia [test c)] indicative of chronic neuropathic pain.
• Rats are injected intra-articularly in one knee joint with Freund's complete adjuvant
• the load that the rat will tolerate on that leg decreases and remains depressed for up to 5 days. This effect is indicative of mechanical hyperalgesia, and is responsive to NSAID's and opiates.
• the compounds of the formula I are administered by injection and preferably orally at doses of from about 3 to 60 microgram/kg animal body weight. The increased load tolerated on the injected side is measured to determine the reversal of hyperalgesia.
• Compound M shows particularly interesting activity on p.o. administration from about 3 to about 60 microgram/kg with a duration of action of about 1 hour. There is no significant difference in response between the doses of 3, 30 and 60 microgram/kg suggesting that the maximum effect had been reached in the range 3 to 30 microgram/kg.
• a local intra-plantar injection of turpentine/paraffin in rat paw (left hind) results in a local inflammatory response and a reduction in the withdrawal threshold (cut-off threshold 340 g) for a mechanical stimulus (paw pressure).
• the compounds of formula I are active at doses from about 1 to 100 microgram/kg p.o. or s.c. administered three days after the injection, further threshold readings being taken 1 and 3 hours later.
• Compound M shows significant activity at doses of 30 and 60 microgram/kg orally, with the maximum effect at 30 microgram/kg. Morphine has an ED 50 value of 1.2 mg/kg s.c. in this test.
• Unilateral partial ligation of the sciatic nerve eliminates fibres throughout the innervation of a paw of a rat.
• the rats develop hyperalgesia to mechanical and thermal stimuli and allodynia of the partially denervated paw without the induction of autotomy.
• the animals are placed in a perspex box on a thin glass plate and a ramp-shaped heat stimulus is applied to the planter surface of a paw. Latency to paw withdrawal is measured.
• the compounds of the formula I are active at doses from about 1 to about 100 microgram/kg injection (s.c. or preferably orally), administered 12 to 15 days after nerve ligation.
• Compound M is particularly active against thermal hyperalgesia.
• Compound M shows significant activity at doses of 30 and 60 microgram/kg, with the maximum effect at 30 microgram/kg.
• the ED 50 is around 60 microgram /kg p.o.
• Morphine has an ED 50 of around 3 mg/kg in this test when administered subcutaneously.
• Clinical trials may be effected as follows:- Subjects suffering from pain, post-operative pain or post herpetic neuralgesia, are administered with compounds of the formula I, especially compound M, i.v. at a dose of from 0.02 to 5 mg. The alleviation of pain is noted.
• the compounds of the invention are therefore useful as analgesics, e.g. against acute or chronic pain, e.g. chronic neuropathic pain.
• the present invention provides a method for the treatment of pain which comprises administering a therapeutically effective amount of a compound of formula I as defined above, to a subject in need of such treatment.
• this invention provides the use of a compound of formula I as defined above as an analgesic in the preparation of medicament suitable for the treatment of pain.
• this invention provides a compound of formula I as defined above for use in the treatment of pain.
• this invention provides an analgesic composition
• an analgesic composition comprising a compound of formula I as defined above as an analgesic in association with a pharmaceutically acceptable carrier or diluent.
• Indications include pain, for example acute pain associated with tissue damage and inflammation (e.g. post operative pain, burn pain, injuries, etc.) chronic inflammatory pain (e.g. arthritis) and chronic neuropathic pain (e.g. diabetic neuropathy, post- herpetic neuralgia, multiple sclerosis, causalgia, etc.).
• tissue damage and inflammation e.g. post operative pain, burn pain, injuries, etc.
• chronic inflammatory pain e.g. arthritis
• chronic neuropathic pain e.g. diabetic neuropathy, post- herpetic neuralgia, multiple sclerosis, causalgia, etc.
• an indicated daily dose is from about 0.1 to about 10 mg, conveniently administered in unit doses from about 0.02 to about 5 mg, and such unit doses may be administered more than once a day, for example 2, 3, 4, 5 or 6 times a day, but preferably 1 or 2 times per day.
• the NSAID ibuprofen has an ED 50 of about 4 mg/kg p.o. and indomethacin 13 mg/kg p.o..
• Compound M is about 300 times more active.
• the preferred dose range is from about 0.1 mg/day to about 10 mg/day, e.g. 3 - 30 microgram/kg for a 70 kg adult, depending on the severity of the indication(s) and frequency of administration.
• Compound M has been shown to have no serious adverse effects in man after single doses up to 5 mg p.o.
• pharmaceutically acceptable encompasses materials suitable for both human and veterinary use.
• the compounds of the invention may be formulated for administration by any suitable route, the preferred route depending upon the disorder for which treatment is required, and preferably in unit dosage form or in a form that a human patient may administer to himself in a single dosage.
• the composition is suitable for oral, rectal, topical, parenteral, intravenous or intramuscular administration.
• compositions of the invention may be in the form of tablets, capsules, sachets, vials, powders, granules, lozenges, suppositories, reconstitutable powders, or liquid preparations such as oral or sterile parenteral solutions or suspensions. Topical formulations are also envisaged where appropriate.
• a composition of the invention is in the form of a unit dose.
• Unit dose presentation forms for oral administration may be tablets and capsules containing 0.02 - 5 mg of a compound of the invention and may contain conventional excipients such as binding agents, for example syrup, acacia, gelatin, sorbitol, tragacanth, or polyvinylpyrrolidone; fillers, for example lactose, sugar, maize-starch, calcium phosphate, sorbitol, or glycine; tabletting lubricants, for example magnesium stearate; disintegrants, for example starch, cross-linked polyvinylpyrrolidone, sodium starch glycollate or microcrystalline cellulose; or pharmaceutically acceptable wetting agents such as sodium lauryl sulphate.
• binding agents for example syrup, acacia, gelatin, sorbitol, tragacanth, or polyvinylpyrrolidone
• fillers for example lactose, sugar, maize-starch, calcium phosphate, sorbitol, or glycine
• the solid oral compositions may be prepared by conventional methods of blending, filling, tabletting or the like. Repeated blending operations may be used to distribute the active agent throughout those compositions employing large quantities of fillers.
• the tablets may be coated according to methods well known in normal pharmaceutical practice, in particular with an enteric coating.
• Oral liquid preparations may be in the form of, for example, emulsions, syrups, or elixirs, or may be presented as a dry product for reconstitution with water or other suitable vehicle before use.
• Such liquid preparations may contain conventional additives such as suspending agents, for example sorbitol, syrup, methyl cellulose, gelatin, hydroxyethylcelluiose, carboxymethylcellulose, aluminium stearate gel, hydrogenated edible fats; emulsifying agents, for example lecithin, sorbitan monooleate, or acacia; non-aqueous vehicles (which may include edible oils), for example almond oil, fractionated coconut oil, oily esters such as esters of glycerine, propylene glycol, or ethyl alcohol; preservatives, for example methyl or propyl p-hydroxybenzoate or sorbic acid; and if desired conventional flavouring or colouring agents.
• suspending agents for example sorbitol, syrup,
• solid oral compositions e.g. tablets and capsules
• solid oral compositions e.g. tablets and capsules
• the tablets are compressed from the capsule granulation of the corresponding dose strength.
• Size #3, yellow capsules are used for the encapsulation of compound M, and the respective formulation for the 0.1 and 5 mg dose strengths are shown below.
• the compounds may also be administered in the form of a sustained release form, in order to provide a prolonged duration of action.
• Conventional drug delivery systems may be used to provide sustained release, for example, coated pellets or push-pull osmotic systems.
• fluid unit dosage forms are prepared utilizing the compound and a sterile vehicle, and, depending on the concentration used, can be either suspended or dissolved in the vehicle.
• the compound can be dissolved in polyethyleneglycol or ethanol and diluted with water for injection and filter sterilized before filling into a suitable vial or ampoule and sealing.
• adjuvants such as a local anaesthetic, a preservative and buffering agents can be dissolved in the vehicle.
• Parenteral suspensions are prepared in substantially the same manner, except that the compound is suspended in the vehicle instead of being dissolved, and sterilization cannot be accomplished by filtration.
• the compound can be sterilized by exposure to ethylene oxide before suspending in the sterile vehicle.
• a surfactant or wetting agent is included in the composition to facilitate uniform distribution of the compound.
• compositions may contain from about 0.1 % to about 99% by weight, preferably from 10 to 60% by weight of the active material, depending on the method of administration.
• Compounds M of the invention may also be administered as a topical formulation in combination with conventional topical excipients.
• Topical formulations may be presented as, for instance, ointments, creams or lotions, impregnated dressings, gels, gel sticks, spray and aerosols, and may contain appropriate conventional additives such as preservatives, solvents to assist drug penetration and emollients in ointments and creams.
• the formulations may contain compatible conventional carriers, such as cream of ointment bases and ethanol or oleyl alcohol for lotions.
• Suitable cream, lotion, gel, ointment, spray or aerosol formulations that may be used for compounds of formula I or a hydrate or an addition product with other solvents or salts thereof, are conventional formulations well known in the art, for example, as described in standard text books of pharmaceutics and cosmetics, such as Harry's Cosmeticology published by Leonard Hill Books, Remington's Pharmaceutical Sciences, and the British and US Pharmacopoeias.
• the present invention provides adenosine-derivatives of formula la
• R.' is (C 4 ⁇ )cycloalkyl substituted by a hydroxy group
• R 2 is as defined above or preferably hydrogen, (C ⁇ alkyl or halogen with an atomic number of 9 to 35, and R, is as defined above.
• halogen with an atomic number of 9 to 35 is fluorine, chlorine or bromine;
• C. alkyl is methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, tert.-butyl, especially methyl;
• C 4 _ ⁇ )cycloalkyl is cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl or cycloctyl, especially cyclohexyl or cyclopentyl.
• R 2 and R 3 possess the significances given above and
• X is chlorine or bromine
• R.' possesses the significances given above.
• the above process is conveniently effected by heating a compound of formula Ha together with a compound of formula Ilia in the presence of a solvent such as dioxane to a temperature of between about 80°C to and 120°C, preferably to boiling temperature.
• a solvent such as dioxane
• X is especially chlorine.
• the compounds of formula Ha, as well as a process for the production thereof, are described in published European Patent Application 269 574.
• a further process for preparation of compounds of formula la comprises treating compounds of formula IVa
• R , R 2 and R 3 possess the significances given above with tetrabutylammonium fluoride trihydrate.
• 6-[(trans)-4-hydroxycyclohexyl]-9-purinyl-2'-0-methyl-3',5'-0-(1 ,1 ,3,3-tetraiso- propyl- disilox-1 ,3-diyl-)-D- ribose used in the above process as starting material may be produced as follows:
• 2gof6-chloro-9-purinyl-2 , -0-methyl-3 , ,5'-0-(1 ,1 ,3,3-tetraisopropyldisilox-1 ,3-diyl)-D- ribose are stirred together with 1.32 g of trans-4-hydroxycyclohexylamine and 1 ,4 ml of N-ethyl-diisopropylamine in 80 ml of dioxane during 6 hours in an oil bath of 105°C. The mixture is subsequently cooled to room temperature, filtered and the filtrate concentrated to dryness.
• the so obtained mixture is eluted on silica gel, first with a mixture of hexane/ethyl acetate 7:3 and then with a mixture of ethyl acetate/methanol 95:5.
• the so purified oily compound has a R, value of 0,3
• Example 8 6-[(1 R,trans)-2-Hydroxycyclohexyl]-2'-0-methyl adenosine
• the compound named in the title is obtained analogously to example 1 when 1 R, trans-2-hydroxycyclohexylamine is used instead of trans-4-hydroxycyclohexylamine.
• the compounds according to the invention have interesting pharmacological properties. They are therefore useful as medicaments.
• compounds according to the invention can be used as such, as hydrates or addition products with organic solvents e.g. methanol, ethanol or ethyl acetate.
• organic solvents e.g. methanol, ethanol or ethyl acetate.
• the most interesting compound of formula la is the compound of Example 1 above which is in the following named compound No. 1.
• the compounds of formula la are of interest as analgesic as described above. They are also of interest for other activities. Among other activities, the compounds according to the invention have anti-hypertensive activity, as indicated from the results of the following investigations:
• the compound of Example 1 is preferred.
• the cardioprotective and analgesic indications are preferred.
• the compounds of formula la are also active in the treatment of arrhythmias as indicated by their adenosine A receptor agonist activity, which is selective via-a-vis their activity against the A2 receptor, as stated e.g. in test a) described hereinafter and for example their capability to prolong conduction through the atrio-ventricular (A-V)node of the heart as indicated in test b) described hereinafter.
• they restore sinus rhythm in supraventricular tachycardias, in particular paroxysmal supraventricular tachycardia, reduce cardiac rate in tachycardic atrial fibrilation and return ventricular tachycardias induced by ⁇ -adrenergic stimulation to normal rhythm.
• the compounds of formula la mimic the effect of "preconditioning", the procedure wherein a brief period of ischaemia renders the heart resistant to infarction from subsequent ischaemia. They are, therefore, useful to protect the heart during unstable angina, against myocardial infarction with or without adjunctive thrombolytic therapy, or ischaemic injury in patients undergoing (e.g.) coronary artery bypass graft surgery, angioplasty, cardiac transplantation or non-cardiac surgery.
• the compounds of formula la are especially useful for administering to subjects prone to myocardial infarctions, e.g. as result of diagnosis or those who have already suffered from a myocardial infarction.
• the compounds of formula la lower further plasma insulin without influencing glucose tolerance. They potentiate the insulin effect to increase glucose uptake and decrease lipolysis in adipose tissue leading to lower plasma free fatty acid concentrations (see test d) described hereinafter). Lower plasma free fatty acids in turn lead to lower triglycerides, which leads to increased HDL cholesterol.
• the insulin sparing effect and/or action to lower free fatty acids makes the compounds of formula la useful for the treatment of Type I diabetes and Type II diabetes i.e. non-insulin dependent diabetes.
• the compounds of formula la lower plasma triglycerides and free fatty acids (see test e) hereinafter) and raise HDL cholesterol and hence, are useful in treating lipid dysfunction, and conditions associated with elevated free fatty acids and triglycerides and where an increase in HDL cholesterol is desirable.
• the compounds of formula la show good metabolic stability.
• the invention provides the use of compounds of formula la for the treatment of, or in the preparation of medicaments suitable for the treatment of, pain.
• this invention provides a compound of formula la as defined above for use in the treatment of pain.
• this invention provides an analgesic composition
• an analgesic composition comprising a compound of formula la as defined above in association with a pharmaceutically acceptable carrier or diluent.
• Pig striatal membranes are prepared as previously described by H. Bruns et al. in Molecular Pharmacology 29 (1986) pages 331-344.
• 3 H-NECA a non-selective adenosine receptor agonist is used to label both A. and A 2 receptors.
• IC 50 values are derived from the displacement curves by weighted non-linear least-square curve fitting to the Langmuir equation and pK D values calculated.
• the compounds of formula la show affinity for the A 1 receptor e.g. at a range of from 1 to 500 nM.
• Adenosine A. receptor activation reduces the incidence of, e.g. paroxysmal supraventricular tachycardia, tachycardic atrial fibrillation and other arrhythmias by slowing conduction through the atrio-ventricular (AV) node of the heart detected by increases in the P-R interval.
• the test method involves recording of ECG changes in conscious adult Rhesus monkeys.
• the compounds of formula la are administered at dosage ranges from about 0.1 mg/kg to about 1000 mg/kg p.o. or 0.03 to 30 mg/kg i.v. in the test referred to by C. Clarke et. al. in The
• Preconditioning (5 minutes of ischaemia followed by 10 minutes of recovery) renders the heart very resistant to infarction from subsequent ischaemia (30 min) and reperfusion (3h).
• the test method is disclosed in the article of G. S. Liu et. al. 1991 - Circulation (84), 1 , 350 - 356 and J. D. Thornton et al. in 1992 - Circulation (85), 659-665.
• the compounds of formula la are administered i.v. at a dose of from about 0.01 to 10 mg/kg to rabbits.
• rabbits given 100 ⁇ g/kg of compound No. 1 were protected to nearly the same extent as endogenous preconditioning from infarction induced by a 30 min. period of coronary occlusion.
• Adipocytes are isolated from the epididymal fat pads of normal chow-fed rats by digestion with collagenase. Cells (final concentration 2% v/v) are pre-incubated with adenosine deaminase (1 U/ml) and with test compounds e.g. compound No. 1 and other additions as indicated to measure glucose transport at 37°C.
• [3- 3 H]Glucose (final concentration, 50 ⁇ M, 0.5 ⁇ Ci/ml) is then added after 30 minutes, and incubation is continued for a further 60 min. Incorporation of radioactivity from [3- 3 H]glucose into cell lipids (a measure of glucose transport) is evaluated by extraction of the cell suspension (0.5 ml) with 5 ml of toluene-based scintillant, followed by liquid scintillation counting; water-soluble metabolites and residual [3- 3 H] glucose remain in the aqueous phase and are not detected.
• Lipolysis is measured in a conventional manner. In one method cells are incubated with and without 1 ⁇ M isoproterenol and then the incubation continued for 60 min. The supernatant is separated from the cells after centrifugation through dinonylphthalate and lipolysis estimated by enzymatic determination of glycerol released into the supernatant.
• the compounds of formula la are active at concentrations from 0.1 to 1000 nM.
• Compound No. 1 suppressed lipolysis in rat adipocytes at concentrations from 0.1 to 100 nM.
• compound No. 1 In the presence of adenosine deaminase (1 U/ml), compound No. 1 has no significant effect on the incorporation of radioactivity from [3- 3 H]glucose into cell lipids in the absence of insulin and only a marginal effect in the presence of a maximally stimulating insulin concentration (8 nM), but significantly increases the stimulation of incorporation of radioactivity in a concentration-dependent manner, at concentrations from 0.1-50 nM insulin. These observations indicate that the compound increases the sensitivity of glucose transport to insulin. In the presence of a near-maximally effective concentration of compound No. 1 , the EC 50 for insulin stimulation of [3- 3 H]glucose incorporation into lipids is decreased between 2 and 3 fold.
• the rats, 2 to 3 months of age, weighing ca. 250 grams are kept in a room at a controlled ambient temperature of 22° C and a 12/12 hour light/dark cycle for seven days.
• rat chow and water are available ad libitum. Following an 18 hour fast, rats (5/group) are given test compounds by gavage in 0.5% CMC. The animals receive 1.0 ml/100 g body wt. Three hours after administration, the rats are anesthetized with C0 2 and blood collected via cardiac puncture-.
• Sera are collected and used for glucose, free fatty acid and ⁇ -hydroxybutyrate determination.
• Free fatty acids are measured by acyl- COA peroxidase calorimetric enzyme assay
• glucose is measured by the glucose oxidase method (YSI Model 27, Yellow Spring, Oh) and ⁇ -hydroxybutyrate is assayed with a ⁇ -hydroxybutyrate dehydrogenase-linked enzyme assay (Sigma Kit 310-A - St. Louis, Mo.).
• the compounds of formula la are active at a dose of from about 1 to about 5000 ⁇ g/kg.
• the doses for lowering free fatty acids (the primary result of the effect on adipocytes) for the compound No. 1 are between 5 and 100 ⁇ g/kg at 2 hr post dose. This results in dose dependent decrease in ⁇ -hydroxybutyrate and blood glucose levels.
• the rats (200-220 g) are fed a high fat diet ad libitum. At fed state, 40 mg of streptozotocin/kg of body weight are injected via the tail vein. One week later, those rats are considered to be diabetic which have fed blood glucose of greater than 200 mg/dl and, following an overnight fast, when given an oral glucose tolerance test, have blood glucose of 40 to 80 mg/dl three hours after the test. Four days later, animals are used in the screen, if fed blood glucose levels are greater than 180 mg/dl. Blood glucose is determined with a YSI Glucose Analyzer. The chronic screen test is carried out as follows:
• the compounds of formula la are also active in tests for mean arterial blood pressure, bradycardia and peripheral vasodilation in anaesthetized rats.
• systolic, diastolic, and mean arterial blood pressure mm Hg; left femoral artery, Statham pressure transducer P 23 Gb
• pulse pressure mm Hg
• heart rate beats/min; triggered from the blood pressure curve
• rate of rise of left ventricular pressure dP/dt max , mm Hg/s; Statham pressure transducer P 23 Gb
• cardiac output ml/min/100 g body weight, thermodilution method, right jugular vein and aorta
• total peripheral resistance dynes s.cm "5 /100 g body weight
• Arterial blood pressure, left ventricular pressure, dP/dt ma ⁇ , heart rate, and electrocardiogram are continuously recorded with a Schwarzer polygraph.
• the parameters are measured 30, 20, 10 and 2 min before administration of the test substance and 1 , 5, 10 and 15 min after injection of the drug into the right femoral vein.
• the compounds of formula la are injected at dosage ranges from about 0.001 to about 10 mg/kg animal body weight.
• Compound No. 1 was tested in cumulative doses of 0.003, 0.010 and 0.03 mg/kg, three animals per dose being used.
• suitable unit doses may be 0.1 to 1000 mg, such as 0.1 to 10, 0.5 to 200, 0.5 to 100 or 0.5 to 10 mg, for example 0.1 , 0.5, 1 , 2, 3, 4 or 5 mg; and such unit doses may be administered more than once a day, for example 2, 3, 4, 5 or 6 times a day, but preferably 1 or 2 times per day, so that the total daily dosage for a 70 kg mammal including humans is in the range of about 0.1 to 1000 mg e.g. per os or 0.003 to
• 300 mg i.v. that is in the range of about 0.001 to 20 mg/kg/day, such as 0.007 to 3, 0.007 to 1.4, 0.007 to 0.14 or 0.01 to 0.5 mg/kg/day, for example 0.01 , 0.02, 0.04, 0.05, 0.06, 0.08, 0.1 or 0.2 mg/kg/day; and such therapy may extend for a number of weeks or months.
• the preferred dose range for all the indications of the invention is from 0.1 mg/day to 10 mg/day for a 70 kg adult.
• non-insulin dependent diabetes and for hypertriglyceridemia the indicated dose for humans is 0.2 to 2 mg per day per os and for treating heart failure and other cardiac disorders 0.2 to 10 mg per day in particular 0.5 to 5 mg/day per os or 0.25 to 5 mg i.v. for arrhythmias e.g. tachycardic atrial fibrillation.
• the compounds of formula la may be formulated for administration by any suitable route, the preferred route depending upon the disorder for which treatment is required, and preferably in unit dosage form or in a form that a human patient may administer to himself in a single dosage.
• the composition is suitable for oral, rectal, topical, parenteral, intravenous or intramuscular administration as described above with respect to the analgesic use.
• Unit dose presentation forms for oral administration may be tablets and capsules containing 0.05-20 mg of a compound of formula la.
• the compounds according to this invention, or a hydrate or an addition product with organic solvents will comprise from about 0.5 to about 20% by weight of the formulation, preferably from about 1 to about 10%, for example 2 to 5%.
• the present invention provides a method of administering N 6 - cyclohexyl-2'-0-methyladenosine to a warm blooded animal in order to produce N 6 - hydroxycyclohexyl-2'-0-methyl adenosine.
• the present invention provides in another aspect N 6 -hydroxycycloalkyl-2'-0-alkyl adenosine in pure form, e.g. greater than 95% pure.
• the exemplified compounds described herein meet this criterion.
• the present invention provides in a further aspect N 6 -hydroxycyclohexyl-2'-0-alkyl adenosine free from cyclohexyl-2'-0-methyl adenosine.
• the present invention provides a process for the production of 2'-0-alkyladenosines which comprises treating adenosine with an appropriate alkyl sulphate in the presence of a phase transfer catalyst.
• the 2'0-alkyl adenosine is any compound defined above.
• the present invention provides a process for the production of compounds of formula I as defined above, which comprises reacting a compound of formula VI
• the alkyl sulphate is di(C 1 ⁇ )alky! sulphate.
• phase transfer catalyst is tetrabutyl ammonium hydrogen sulphate.
• reaction is effected in a non-polar solvent, e.g. as described hereinafter.
• a group of compounds of the formula I is defined wherein R. is (C M )cycloalkyl, R 2 is hydrogen or (Chalky., and R 3 is (C, ⁇ )alkyl.
• the preparation of compounds of formula I typically involved six steps, which include the preparation of 2',3',5'-triacetylinosine; chlorination and hydrolysis to 6-chloro-9- ⁇ -D-ribofuranosyl-9H-purine; protection of the 3'-0- and 5'-0-positions with tetraisopropyldisiloxane (TIPDS-CL 2 ); 2'-0-alkylation and purification by silica gel chromatography; deprotection of the 3'-0- and 5'-0-positions; and reaction with R ⁇ H j , and recrystallization to obtain the compound of formula I.
• TIPDS-CL 2 tetraisopropyldisiloxane
• the compounds of formula I can be prepared in good yields and purity without the need for 3'-0- and 5'-0-disiloxane protection and deprotection or for silica gel chromatography.
• the applicants have found that the inosine-2',3,'5'-triacetate starting material of the prior art process may be advantageously replaced with the lipophilic inosine-2',3',5'-tripropionate, which permits the doubling of throughput and the elimination of undesirable pyridine solvent.
• R 1 R 2 and R 3 are as defined above.
• the compounds of formula I may be prepared by reacting a basic aqueous solution of a compound of formula (VI) with a di-(C 1 ,)alkyl sulfate in the presence of tetrabutylammonium hydrogen sulfate and an organic water-immiscible solvent.
• the base used to prepare the basic aqueous solution is preferably an alkali metal hydroxide, such as sodium hydroxide or potassium hydroxide.
• the solvent can be any water immiscible or essentially immiscible solvent in which the compound of formula I is soluble, such as methylene dichloride or t-amyl alcohol. It is also preferred that the reaction be run at temperatures between about -20°C to about
• the crude compound of formula I is isolated by evaporation followed by stirring with a 1 : 1 mixture of water and inert solvent, preferably toluene, at room temperature for 5 to 7 hours, then filtering and drying. Pure compound may be obtained by fractional crystallization, preferably by:
• step 2) repeating the recrystallization procedure of step 1) but heating at about 65°C before cooling to room temperature;
• R. and R 2 are as defined above.
• the compounds of formula VI may be prepared by acylating inosine of formula VII with propionic anhydride to form the 2'-0-, 3'-0-, 5'-0-protected compound of formula VIII; halogenating the compound of formula VIII with thionyl chloride to form the intermediate of formula IX; and simultaneously aminating and hydrolyzing the compound of formula IX.
• Acylation of inosine is preferably carried out in a mixture of toluene, tributylamine, and 4-dimethylaminopyridine at a temperature of from 100° to 110°C over a period of 4 to 5 hours.
• the compound of formula VIII is isolated by precipitation with heptane.
• Halogenation of the compound of formula VIII is preferably carried out in a mixture of toluene and N,N-dimethylformamide at a temperature of from about 60° to 70°C over a period of 3 to 4 hours, and the solution of the compound of formula IX may be used after washing with water and brine.
• Amination and simultaneous hydrolysis of the intermediate of formula IX is preferably effected by adding the amine R 1 NH 2 and reacting at a temperature of from 100° to 110°C over a period of 15 to 20 hours.
• the compound of formula VI is isolated by filtration at room temperature and purified by recrystallization.
• a mixture of 271.2 g of inosine, 966 ml of tributylamine, 3.30 g of 4-dimethylaminopyridine and 600 ml of toluene is heated to an internal temperature of 104-105°C; and over a period of 35 minutes, 453 ml of propionic anhydride are added at a rate which maintains the internal temperature between 104-105°C. After stirring the mixture at this temperature for an additional 4 hours, the mixture is cooled to 5-10°C with an ice bath; and 1000 ml of heptane are added. The resulting suspension is stirred at room temperature (20-22°C) for 30 minutes and then filtered e.g. on a Buchner funnel.
• a mixture of 270.6 g of inosine-2',3',5'-tripropionate, 240 ml of N,N-dimethylformamide and 600 ml of toluene is heated to 65°C; and over a period of 1 hour, 67.84 ml of thionyl chloride are added at a rate which maintains the internal temperature between 62-65°C.
• the mixture is stirred at this temperature for an additional 2.5 hours and then cooled to 10°C with an ice bath.
• the suspension is stirred for 15-20 min and 175 ml of tert-butyl ethyl ether are added. After stirring for 5 min, the suspension is cooled in an ice bath and stirred for an additional 15 minutes. This suspension is filtered in a Buchner funnel and washed with a total of
• the filtered solid is dried at 45-50°C (30-35 mbar) for 14 hours to yield 130 g of N 6 -cyclohexyladenosine as a white solid (m.p. 185-187°C; yield 60.0%).
• a suspension of 94.33 g of N 6 -cyclohexyladenosine and 720 g of 5% aqueous sodium hydroxide is stirred at room temperature (24-25°C) until all of the solids are dissolved (approximately 5 min.).
• 850 ml. of dichloromethane and 5.5 g of tetrabutylammonium hydrogen sulfate are added followed by 61.3 g of dimethyl sulfate over 5-10 minutes, while maintaining the internal temperature between 24-25°C.
• the addition funnel is washed with an additional 50 ml of dichloromethane which is added to the reaction vessel.
• a mixture of the above-mentioned crude material and 2470 ml of toluene is stirred at room temperature for 10 minutes and then 2470 ml of water are added over a period of 22 min.
• the resulting suspension is stirred at room temperature for an additional 6 hours and solids are collected by filtration e.g. on a Buchner funnel with suction. After washing the solids with 114 ml of toluene and a total of 285 ml. of water in three equal portions of 95 ml each, the solids are dried at 48-50°C (25 mm Hg) overnight (14 hours) to yield 53.0 g of a white solid. A suspension of this solid in 397 ml.
• toluene is heated to 80°C with stirring to form a clear solution, which is cooled to 56°C over 45 min and seeded with 10 mg of pure product.
• the mixture is stirred at 55-56°C for 45 minutes and then cooled to room temperature over 1 hour. After stirring at this temperature for an additional 1 hour the solid are collected by filtration e.g. on a Buchner funnel with suction.
• the solids are washed with a total of 75 ml. of toluene in three equal portions of 25 ml. each to yield 60 g of a white solid.
• a suspension of this solid in 159 ml of toluene is again heated to 80°C, and the above seeding procedure is carried out at 65° to 66°C.
• the solids are collected by filtration on a Buchner funnel with suction and washed with a total of 57 ml. of 1 :2.36 mixture (v/v) of 100% ethanol and water in three equal portions of 19 ml each.
• the solids are dried at room temperature (29 mm Hg) to yield 62.2 g of product as a white solid in 1.5 hydrate form (m.p. 88°-91°C; yield
• the process of this invention is more economic in time and cost than hitherto known processes.
• the process provides a convenient method for selective 2'-0- methylation of N 6 -cyclohexyladenosine under phase-transfer conditions, and a method of purification of the 2'-0-methyl derivative. Expensive protecting groups and the use of chromatography are avoided.

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• 1995
  • 1995-05-09 SK SK1460-96A patent/SK146096A3/sk unknown
  • 1995-05-09 EP EP95919777A patent/EP0759925A1/en not_active Withdrawn
  • 1995-05-09 AU AU25461/95A patent/AU2546195A/en not_active Abandoned
  • 1995-05-09 HU HU9603105A patent/HUT75338A/hu unknown
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  • 1995-05-09 JP JP7529185A patent/JPH09512823A/ja active Pending
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• 1996
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