CN1094046A

CN1094046A - The xanthine derivative that replaces

Info

Publication number: CN1094046A
Application number: CN93107586A
Authority: CN
Inventors: A·E·芬韦克
Original assignee: SmithKline Beecham Ltd
Current assignee: SmithKline Beecham Ltd
Priority date: 1992-05-21
Filing date: 1993-05-20
Publication date: 1994-10-26
Also published as: WO1993023401A1; CA2136196A1; EP0641344A1; MX9302957A; AU4081493A; GB9210839D0; ZA933494B; JPH07506591A; TW277062B

Abstract

The compound of a kind of formula (I) or the acceptable salt of its suitable medicine, formula (I) is: R wherein ¹And R ²Difference expression (a) part:

-(CH ₂) _m-A (a)

Wherein m represents 0 or integer 1,2 or 3, and A represents the cyclic hydrocarbon group that replaces or do not replace; R ³Expression hydrogen, replacement or the alkyl that does not replace, or the aralkyl that in aryl moiety, replaces or do not replace; And R ⁴Expression hydrogen, alkyl or alkyl-carbonyl; Prepare this compound and the method and the purposes of this compound in treatment that contain the pharmaceutical composition of this compound.

Description

The xanthine derivative that replaces

The present invention relates to the preparation method of some novel cpds and such compound thereof, also related to the pharmaceutical composition and these compounds or the application of composition in medical science that contain these compounds with pharmacological activity.

Molecular pharmacology (Molecular pharmacology), 6 volumes, No.6,1970, P597～603) disclosed 1,3-dimethyl-8-nitro-xanthine.This compound has lipolytic activity.Ann Chim, 47,362-365(1957) disclosed 1,3-dimethyl-8-amino-xanthine and preparation method thereof, this compound does not have pharmacology effectiveness.Drug Res, 27(1) Nr.19,1977,4-14 page or leaf, Van K.H.Klingler disclosed some 1, the xanthine that 3-dimethyl-8-replaces, they the styroyl aminoalkyl xanthic synthetic in conduct intermediate independently.Drug Res, 31(11), Nr.12,1981, people such as R.G.Werner, the 2044-2048 page or leaf disclosed some 1, the xanthine that 3-dimethyl-8-replaces, these compounds do not have pharmacological activity.

European patent application (publication number 0369744) also disclosed some 1,3-or 1,3,7-8-H cycloalkyl alkylidene group xanthine, relate in addition they as bronchodilator in the application of treatment in the asthma.

It has been found that some new 8-substituted xanthines have the activity as phosphodiesterase inhibitor.

These compounds show it is the good inhibitor of inducing blood eosinocyte quantity to increase.Therefore, these compounds can be used for treating and/or preventing by eosinocyte quantity and increase and the illness such as the asthma that cause, and allergic conditions such as the urticaria, eczema and the rhinitis that are caused by specific reaction.

These compounds also show the activity with bronchodilator, therefore can be used for treating illness such as the reversible airway obstruction and the asthma of respiratory organs system.

These compounds also have the protective effect that stops the influence of cerebrum metabolism restraining effect.Described compound has improved the recovery behind data acquisition or the instantaneous preceding cerebral ischaemia, thereby they can be used to treat the brain vascular and neuronic sex change illness, above-mentioned illness and study, remember and comprise brain aging, be subjected to the infarct dementia, Alzheimer type senile dementia, with memory impairment follow aging relevant.And they also can be used to treatment by Parkinson's (some illnesss that the disease of Parkinson ' s) causes.

These compounds also show the activity with neuroprotective; thereby they comprise because the prevention of cardiac's dislocation in prevention, apoplexy and the ischemic case of the cerebral ischemia that causes causes neuronic sex change and in the illness that produces and as by surgery and/or these cerebrum ischemia cases of during baby due, causing after the neuronic sex change that caused and be useful in the illness that produces.In addition, showing with this compounds for treating, is useful for treatment by the functional illness that brain function imbalance after the local asphyxia is caused.

These compounds also are effective to improving oxygen tension in the local ischemic skeletal muscle.This performance makes nutrition blood stream increase by local asphyxia marrow flesh, shows also that therefore The compounds of this invention can be used as the medicament of treatment peripheral vascular disease such as intermittent claudication.

Compound of the present invention also can be used as the inhibitor that produces tumour necrosis factor (Tumor Necrosis Factor) in the organism of living, thereby they can be used to treat the disease of following excessive or irregular TNF to produce, comprise rheumatoid arthritis, similar rheumatism spondylitis, osteoarthritis, gouty arthritis and other arthritis disease; Sepsis, septic shock, the endogenous toxic material shock, the Gram-negative sepsis, the shock of poisoning syndromes, grownup's respiratory distress syndrome, encephalic malaria, chronic pneumonia, silicosis, lung's sarcoidosis, bone absorpting disease, fusion damage (reperfusien injury) graft is to host's reaction again, allograft rejection, owing to infect the fever and myalgia such as the influenza that cause, evil matter disease secondary infection or malignancy, evil matter disease, secondary obtain immunity loss syndromes (AIDS), AIDS, the relevant multiple disease of ARC(AIDS), keloid forms, scar tissue forms, Crow engler (the disease of crohn ' s), ulcerative colitis or pyresis.Compound of the present invention also is used for the treatment of because infection produces the viral infection of TNF, or by the disease of The compounds of this invention treatment to the inhibition sensitivity, for example realizes by being reduced directly or indirectly to duplicate by some this discovery compounds.Such virus comprises as HIV-1, HIV-2, HIV-3, cytomegalovirus (CMV), influenza virus, adenovirus and simplexvirus group, for example varicella zoster virus and herpes simplex virus, and be not limited to these virus.

Comprise veterinary treatment the above-mentioned course of treatment, by the TNF that virus infection causes, for example comprise cat family immune deficiency virus (FIV) or other retrovirus infection such as equine infectious anemia virus, caprine arthritis virus, visna virus, sheep chronic pneumonia (maedi) virus and other slow virus (lentiviruses) particularly including treatment.

These compounds also can be used for the treatment of the mankind or other mammalian skin hyperplasia diseases.

Therefore, the invention provides a kind of salt of accepting as compound or its suitable medicine of formula I, formula I is:

R wherein ¹And R ²Difference expression (a) part:

-（CH ₂） _m-A （a）

Wherein m represents 0 or integer 1,2,3, and A represents a cyclic hydrocarbon group that replaces or do not replace;

R ³Expression hydrogen, replacement or the alkyl that does not replace or an aralkyl that is substituted or does not replace at aryl moiety;

R ⁴Expression hydrogen, alkyl or alkyl-carbonyl.

Proper A does not replace, and well, A represents the C that replaces or do not replace _3-8Cycloalkyl, especially C _3-6Cycloalkyl.

Especially, A represents cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl a replacement or that preferably do not replace.

Well, A representative ring propyl group or cyclobutyl.

Preferably, A representative ring propyl group.

As R wherein ³The expression do not replace alkyl the time, suitable example comprises methyl.

As R wherein ³When representing the alkyl of replacement, suitable example comprises alkoxyl-methyl such as methoxyl methyl.

Suitably, R ³Expression aralkyl, for example benzyl that replaces or do not replace at aryl moiety.

Work as R ³When being benzyl, example comprises benzyl that does not replace or the benzyl that is replaced by methoxyl group in phenyl, and special example comprises the 4-methoxy-benzyl.

Suitable R ⁴Expression hydrogen.

Suitable R ⁴The expression alkyl-carbonyl.

Suitable alkyl-carbonyl is C _1-4Alkyl-carbonyl such as ethanoyl.

Suitable drug acceptable salt is acceptable basic salt of medicine and the acceptable acid salt of medicine.The suitable medicine of formula I compound can be accepted the 7-N subsalt that basic salt comprises metal-salt, comprises the salt of an alkali metal salt such as sodium salt or organic amine salt such as 1.

The acid salt that the formula I compound is suitable comprises the acceptable inorganic salt of medicine such as the acceptable organic acid addition salt of vitriol, nitrate, phosphoric acid salt, borate, oxyhydroxide, hydrobromide and medicine such as acetate, tartrate, maleate, Citrate trianion, succinate, benzoate, ascorbic acid, metilsulfate, alpha-ketoglutarate, α-glycerophosphate and Cori ester salt.Preferred acid salt is a hydrochloride salt.

Utilize ordinary method to prepare the salt that the formula I compound medicine is accepted.

Terminology used here " cyclic hydrocarbon group " comprises monocycle and condensed ring, and it is no more than 8 to contain carbonatoms in each ring of alicyclic ring, and suitable is no more than 6 carbon atoms, for example 3,4,5 or 6 carbon atoms.

For any cyclic hydrocarbon group, suitable selectivity substituting group comprises a C _1-6Alkyl or a halogen atom.

No matter used term " aryl " is to use separately or the part (as in aralkyl) of the other group of conduct, all comprise phenyl and the naphthyl of selecting no more than 5 replacements, preferably be no more than 3, replacement is selected from halogen, alkyl, phenyl, alkoxyl group, haloalkyl, hydroxyl, amino, nitro, carboxyl, alkoxy carbonyl, alkoxy carbonyl alkyl, alkoxy carbonyl oxygen or alkyl-carbonyl.For any phenylene, the substituting group of selection comprises no more than 3 substituting group relevant with described aryl.

For the aryl moiety of any aralkyl, suitable selectivity substituting group comprises the substituting group of above-mentioned relevant " aryl ", particularly including alkoxyl group such as methoxyl group.

No matter terminology used here " alkyl " is to use separately or as the part (as in alkyl-carbonyl) of other group, all comprise the straight chain and the branched-chain alkyl that contain 1～12 carbon atom, preferably contain 1～6 carbon atom, for example methyl, ethyl, propyl group or butyl.For any alkyl, suitable selectivity substituting group comprises and is no more than 5 the substituting group relevant with above-mentioned aryl, preferred no more than 3 substituting groups.

Here used noun " skin hyperplasia disease " is meant and optimum and virulent skin hyperplasia disease it is characterized in that following incomplete tissue differentiation, in epidermis, corium or the cell fission quickened in this place's appurtenant.These diseases comprise: psoriasis, different keratosis of answering dermatitis, non-specific dermatitis, initial stage stimulation contact dermatitis, allergy contact dermatitis, skin substrate and squamous cell carcinoma, layer sauriasis, the excessive disease of keratinization of epidermis, the preceding Exposure to Sunlight of deterioration to cause.Atopic dermatitis and the mange of non-pernicious keratosis, acne, human seborrheic dermatitis and domestic animal.

The compound of formula I is preferably with the acceptable form of medicine.In addition, the acceptable form of medicine is meant the acceptable scale of medicine, and it does not comprise conventional medicated premix such as thinner and carrier, does not also contain the toxicant of any common dosage.Get rid of conventional medicated premix, the acceptable scale of this medicine generally is at least 50%, and is preferred 75%, more preferably is 90%, and more preferably 95%.

The present invention has further proposed a kind of method for preparing the formula I compound, and this method comprises the compound reaction of the compound and the formula III of formula II, and formula II is:

R wherein ^1aExpression is as the R of formula I definition ¹Or one can change R into ¹Group, R ^2aExpression is as the R of formula I definition ²Or one can change R into ²Group, R ^3aExpression is as the R of formula I definition ³Or one can change R into ³Group; Formula III is:

R wherein ⁵Expression hydroxyl protecting group, L ¹Represent a leavings group; If desired, can carry out selectivity step below one or several:

(ⅰ) remove arbitrary blocking group;

(ⅱ) arbitrary R ^1aBe converted into R ¹And/or arbitrary R ^2aBe converted into R ²And/or arbitrary R ^3aBe converted into R ³;

(ⅲ) compound of formula I is changed into the compound of another formula I;

(ⅳ) compound of formula I is converted into the acceptable salt of its medicine.

Suitable leavings group L ¹Be a halogen atom, iodine atom especially.

The reaction of formula II and formula III compound can be finished under the alkylation conditions of routine, for example in aprotic solvent as glycol dimethyl ether, dimethyl formamide or tetrahydrofuran (THF), arbitrary temperature of suitable finished product rate of formation can be provided, for example 0 ℃～100 ℃, be generally 40 ℃～80 ℃, as 60 ℃; And preferably in rare gas element such as nitrogen.

The 8-amino group that compound (II) is suitable is active form, is the form with ionic form such as salt well, for example by the form of formula II compound and basic metal base such as uncle-alkali metal base that Ding oxygen nak response provides.

The compound of formula II can utilize the method preparation that is documented in the european patent application (publication number 0389282).

The compound of formula III is a known compound, and perhaps they can be by the method preparation for preparing known compound, for example at Tetrahedron(1990), those methods of discussing in 46,6903.

The form that a compound of formula I changes another compound of formula I into comprises a R ⁴Base changes another R into ⁴Base is for example R ⁴The hydrolysis compound that is an alkyl-carbonyl is transformed into R ⁴It is the formula I compound of a hydrogen atom.

In described conversion process, suitably utilize suitable ordinary method, and, in the said hydrolyzed process, what also use is common hydrolysising condition, for example in ethanolic soln such as methyl alcohol, utilizes the weak base effect to finish the hydrolysis of alkyl-carbonyl, preferably use salt of wormwood, this is normally in envrionment temperature.

For R ^1aAnd R ^2a, suitable form comprises R respectively ¹And R ²Or nitrogen-protecting group group is as benzyl, nitrobenzyl or trimethoxy benzyl.For R ^3a, suitable form comprises R ³

When R is the aralkyl that replaces or do not replace, suitable R ^1aAnd R ^2aBeing illustrated in does not influence R ³Situation under the nitrogen-protecting group that can insert and remove, for example trimethyl silyl.

R ³When being the aralkyl that replaces or do not replace, preferred R ^1aBe R ¹, R ^2aBe R ²

Work as R ^1a, R ^2aOr R ^3aExpression is different from R respectively ¹, R ²Or R ³The time, above-mentioned R ^1aBe converted into R ¹, R ^2aBe converted into R ², R ^3aBe converted into R ³, these conversion processes can be carried out with suitable ordinary method.R for example ^1a(or R ^2a) when being expressed as a nitrogen-protecting group such as benzyl, just can remove this protecting group with suitable ordinary method, suitable method is catalytic hydrogenation for example, and a compound reaction with formula IV obtains product then, and formula IV is:

Wherein the definition of A and m is suc as formula definition accordingly in (I A), and X represents a leavings group, for example halogenide such as bromide or iodide.

The protection of any reactive group or atom such as xanthine nitrogen-atoms can be in aforesaid method suitably step carry out.Suitable protecting group comprises in the prior art the specific bases of protection that those are commonly used or the group of atom, and for example for the xanthine nitrogen-atoms, its suitable protecting group is alkyl silyl, particularly trimethyl silyl or tert-butyl dimetylsilyl.

Utilize suitable ordinary method can prepare and remove protecting group: the method that for example prepares alkyl silyl blocking group can be handled the compound of formula II with suitable alkyl silyl halides; for example for the trimethyl silicon based halogenide of trimethyl silyl; for the tert-butyl dimetylsilyl; with tert-butyl dimetylsilyl halogenide; by in suitable solvent, for example tetrahydrofuran (THF) is handled with the tert-butyl Neutral ammonium fluoride at ambient temperature and can be removed the silyl blocking group easily.

The present invention's compound with treatment characteristic above-mentioned is: by formula I compound or acceptable salt of its suitable medicine and/or the acceptable solvate of its medicine as the active treatment material provided by the invention.

Therefore, the invention provides compound or the acceptable salt of its suitable medicine and/or the acceptable solvate of its medicine of formula I, allergic conditions such as urticaria, eczema and rhinitis that they are used for treating and/or preventing the illness following the eosinocyte number to increase and produce such as asthma and are caused by specific reaction.

On the other hand, the present invention also provides a kind of compound or its acceptable salt of a kind of suitable medicine and/or the acceptable solvate of its a kind of medicine as a kind of formula I of phosphodiesterase inhibitor.

As mentioned above, the special one side of the present invention provides a kind of compound or its acceptable salt of a kind of suitable medicine and/or acceptable solvate of a kind of medicine that is used for the treatment of the formula I of respiratory organs system illness such as reversible airway obstruction and asthma.

Another special aspect, a kind of formula I compound provided by the invention or its acceptable salt of a kind of suitable medicine or the acceptable solvate of its a kind of medicine, they are used for treatment above-mentioned, cerebral blood vessel and neuronal degeneration illness, peripheral vascular disease or skin hyperplasia disease that for example concomitant learning, memory and cognitive dysfunction produce, or be used to prevent to cause the illness of following neuronal degeneration to produce by local asphyxia.

On the other hand, the present invention also provides as the compound or acceptable salt of its suitable medicine and/or the acceptable solvate of its medicine that produce the formula I of tumour necrosis factor (TNF) inhibitor in the organism of living.

Formula I compound particularly of the present invention or the acceptable salt of its suitable medicine and/or the acceptable solvate of its medicine are used for the treatment of and/or prevent owing to excessive or irregular TNF produces the disease that causes.

The disease that is used for excessive or irregular TNF generation and causes comprises rheumatoid arthritis, similar rheumatism spondylitis, osteoarthritis, gouty arthritis and other arthritis; Sepsis, septic shock, the endogenous toxic material shock, the Gram-negative sepsis, the shock of poisoning syndromes, grownup's respiratory distress syndrome, cerebral malaria, chronic pneumonia, silicosis, sarcoidosis of lung, bone absorpting disease, damage (reperfusion injury) is closed in remelting, graft is to host's reaction, the repulsion of allograft, owing to infect the fever and myalgia such as the influenza that cause, secondary infection or malignancy emaciation, emaciation, secondary acquired immunodeficiency syndrome (AIDS), AIDS, the multiple disease that ARC(AIDS is relevant), keloid forms, scar tissue forms, Crow engler (the disease of crohn ' s), ulcer colonitis or pyresis.

On the other hand, formula I compound provided by the invention or the acceptable salt of its suitable medicine or the acceptable solvate of its medicine are used for the treatment of and/or prevent owing to infect the viral infection that causes generation TNF, or, duplicate as reducing directly or indirectly by compound of the present invention to those diseases of inhibition sensitivity.These viruses comprise as HIV-1, HIV-2 and HIV-3, cytomegalovirus (CMV), influenza virus, adenovirus and bleb papova, for example varicella zoster virus and herpes simplex virus and be not limited to these virus.

The compound of formula I or the acceptable salt of its suitable medicine and/or the acceptable solvate of its medicine can directly be offerd medicine, or preferred pharmaceutical composition as a kind of drug acceptable carrier.

Therefore, pharmaceutical composition provided by the invention contains formula I compound or the acceptable salt of its suitable medicine and/or the acceptable solvate of its medicine and a kind of medicine acceptable carrier.

The active compound that is used for introducing body can be by any suitable approach preparation, and preferred approach depends on the disease that will treat, and preferred with unitary dose form or can be with patient to the form of oneself executing individually dosed medicine.Useful is, composition of the present invention be suitable for mouth, rectum, part, injection, intravenously or muscle administration or pass through respiratory tract administration.The preparation of design can make active ingredient discharge at leisure.

The form of the present composition can be: tablet, capsule, sachet, small bottle packing, pulvis, particle, lozenge, suppository, can reconstituted powder or liquid preparation such as oral liquid or aseptic injection solution or suspension.The prescription of topical also can act on suitable position.In order to satisfy quantitative taking medicine, the present composition is preferably with the form of unitary dose.

As oral medicine, the appearance forrns of unitary dose can be tablet and capsule, and can contain conventional vehicle, for example tackiness agent such as syrup, gum arabic, gelatin; Sorbyl alcohol, tragacanth gum or polyvinylpyrrolidone; Weighting agent such as lactose, sucrose, W-Gum, calcium phosphate, sorbyl alcohol or glycine, sheet lubricant such as Magnesium Stearate; Decomposition agent such as starch, polyvinylpyrrolidone, Explotab or Microcrystalline Cellulose; Or acceptable wetting agent of medicine such as sodium lauryl sulphate.

The ordinary method of preparation Peroral solid dosage form drug composition is: mixing, filling, film-making etc.Repeating married operation can make active agents be distributed in the whole composition that contains a large amount of weighting agents.

Above-mentioned operation is generally prior art certainly, and in general medicinal practice, tablet medicine can particularly be wrapped enteric coating by the currently known methods dressing.

The form of oral liquid for example can be: milk sap, syrup or elixir, or can be used as dry preparation and carry out recombinant with water or other suitable carriers before use.These liquid preparations can contain general additive, for example suspension agent such as Sorbitol Powder, syrup, methylcellulose gum, gelatin, Natvosol, carboxymethyl cellulose, aluminium stearate gel, hydrogenation edible fat; Emulsifying agent such as Yelkin TTS, polyoxyethylene-sorbitan mono-oleate or gum arabic; Water-free carrier (can comprise edible oil) is as Prunus amygdalus oil, fractionated coconut oil, oily ester such as glyceryl ester, propylene glycol or ethanol; Sanitas such as methyl or propyl group be right-hydroxy benzoate or sorbyl alcohol; Add conventional essence or pigment on demand.

For making medicine be given to respiratory tract, suitable composition forms also can be with the atomizer effect smell agent or smoke substance or solution, or the fine-powder that blows into can use separately or mixes with inert support such as lactose.In this case, the suitable diameter of active ingredient composition granule is less than 50 microns, and for example 0.1～50 micron, preferably less than 10 microns, as 1-10 micron, 1～5 micron or 2-5 micron.In appropriate circumstances, can comprise a spot of antasthmatic and bronchiectasis warp, for example sympathomimetic amine and isoprene suprarenin, neoisuprel, salbutamol, phyenlephrinium and ephedrine; Corticosteroid is as prednisolone and suprarenal gland energizer such as ACTH.

As administered parenterally, utilize this compound and a kind of sterile carrier to prepare the liquid unit dosage, and be decided by used concentration, compound can suspend or be dissolved in the carrier.In preparation solution, this compound dissolves in the water as injection, and sterilising filtration, sealing then before pack into suitable medicine bottle or ampoule.

Advantageously, the component that is dissolved in the carrier has auxiliary such as local anesthetic, sanitas and buffer reagent.For enhanced stability, composition can carry out freezing before the medicine bottle of packing into and go down to dewater in vacuum.The preparation of the suspension of administered parenterally except compound is to be suspended in the carrier to be dissolved in wherein with replacement, and can not be finished germ-resistant purpose with filter method basically by above-mentioned similar methods.Compound can be before being suspended in sterile carrier carries out sterilization by the method that is exposed among the oxyethane.Useful is also to comprise a kind of tensio-active agent or wetting agent in said composition, in order that impel compound to distribute equably.

Content of active substance is 0.1%～99%wt in the said composition, is preferably 10～60%wt, and this depends on the method for administration.

The local prescription that the compound of formula I or the acceptable salt of its suitable medicine and/or the acceptable solvate of its medicine also can be used as with common partial mixed with excipients is introduced in the body.

The partial prescription such as the ointment that can be provided, emulsifiable paste or lotion, the dressing that infiltrated, gel, gel paste (gel sticks), spray and smoke substance, and can contain suitable typical additives such as sanitas, solvent in order that make medicine infiltration and lubricant in ointment and emulsifiable paste.This prescription also can contain compatible body commonly used, for example emulsifiable paste or ointment base and as the ethanol or the oleyl alcohol of lotion.

Compound or the acceptable salt of its suitable medicine for formula I, spendable suitable emulsifiable paste, lotion, gel, gel subsides, ointment, spray or smoke substance prescription are prescriptions commonly used in the prior art, for example they are documented in the common textbook of medicine and makeup, publish by Leonard Hill Books as Harry ' s Cosmeticology(), Remington ' s pharmaceutical Sciences and the British and US Pharmacopoeias.

The compound of the formula I that adapts or the acceptable salt of its suitable medicine contain the about 0.5～20% of formulation weight, and is preferred about 1～10%, and for example 2～5%.

The compound dosage that the present invention is used for the treatment of can change with common situation, promptly changes with the seriousness of illness, patient's body weight and the corresponding significant quantity of compound.But as common guide, suitable unitary dose can be 0.1～100mg, 0.5～200mg for example, and 0.5～100mg or 0.5～10mg are as 0.5,1,2,3,4 or 5mg; More than this unitary dose can be taken once in one day, every day 2,3,4,5 or 6 times for example, but take 1 or 2 preferred every day, in order that make grownup every day of 70Kg body weight total dosage in the scope of about 0.1～1000mg, promptly about 0.001～20mg/Kg/ days, for example 0.007～3,0.007～1.4,0.007～0.14 or 0.01～0.5mg/Kg/ days, as 0.01,0.02,0.04,0.05,0.06,0.08,0.1 or 0.2mg/Kg/ days; Such therapy can continue a few weeks longer or some months.

Terminology used here " medicine is acceptable " comprises the material that is suitable for for the mankind and animal doctor's effect.Do not find that in above-mentioned dosage range the formula I compound has any toxicity.

The present invention is set forth by following pharmacology data and embodiment, and following preparation method has illustrated by intermediate preparation novel cpd formula I.

Embodiment 1

8-[4-acetoxyl-3-(acetoxyl methyl) butyl ammonia]-1,3-two (cyclopropyl methyl)-7-(4-methoxy-benzyl) xanthine

Under 60 ℃, condition of nitrogen gas, uncle-Ding oxygen potassium (0.35g, 3.13mmol) join the 8-amino-1 in the glycol dimethyl ether (DME 10ml), 3-two (cyclopropyl methyl)-7-(4-methoxy-benzyl) (0.99g is in solution 2.5mmol) for xanthine.After 3 hours, within 5 minutes, add DME(3ml at leisure) in 4-acetoxyl-3-(acetoxyl methyl) butyl iodide (1.69g, solution 5.4mmol).Stir after 18 hours, cool off this mixture, pour ethyl acetate into, washing organic solution, dry, evaporation.On silica, obtain 8-[4-acetoxyl-(3-acetoxyl methyl) butyl-amino with chromatography (acetone/hexane 1: 7)]-1,3-two (cyclopropyl methyl)-7-(4-methoxy-benzyl)-xanthine (0.63g, 43%), mp129～129.5 ℃.

δ (CDCl ₃) 0.43-0.50(8H, m), 1.26-1.38(2H, m), 1.58(2H, m), 1.91(1H, m), 2.05(6H, s), 3.47(2H, m), 3.79(3H, s), 3.90(2H, d, J=3.9Hz), 3.93(2H, d, J=3.9Hz), 4.03(4H, m), 4.30(1H, t, J=6.0Hz), 5.29(2H, s), 6.88(2H, d, J=8.5Hz) and 7.22(2H, d, J=8.5Hz);

V _Max(KBr) 3272(m), 1742(s), 1693(s), 1651(s), 1617(s), 1566(s), 1250(s), 1240(s), 1221(s) and 1040(s) cm ^-1;

M/e(FAB) 121(100%), 145(34), 105(27), 582(MH ⁺, 25) and 604(MNa ⁺, 15);

Find C, 61.78; H, 6.89; N, 11.82; C ₃₀H ₃₉N ₅O ₇Satisfy C, 61.94; H, 6.76; N, 12.04%

The initial substance identical with actual sample (0.41g, 42%) arranged.

Embodiment 2

1,3-two (cyclopropyl methyl)-8-[4-hydroxyl-3-(methylol) butyl amino]-the 7-(4-methoxy-benzyl) xanthine

At room temperature, in methyl alcohol (8ml), stir 5 hours 8-[4-acetoxyl-3-(acetoxyl methyl) butyl amino]-1,3-two (cyclopropyl methyl)-7-(4-methoxy-benzyl) xanthine (0.22g, 0.37mmol) and salt of wormwood (0.005g, 0.037mmol).This mixture neutralizes with cHCl, and under reduced pressure removes this solvent.(hexane/acetone gradient) obtains 1 with resistates (250ml) chromatography on silicon face, 3-two (cyclopropyl methyl)-8-[4-hydroxyl-3-(methylol) butyl amino]-the 7-(4-methoxy-benzyl) xanthine (0.11g, 59%), mp.141-2 ℃.

δ (CDCl ₃) 0.39-0.49(8H, m), 1.28-1.38(2H, m), 1.61-1.65(3H, m), 2.29(2H, t, J=4.5Hz), 3.46(2H, m), 3.68(4H, br s), 3.80(3H, s), 3.92(4H, the overlapping d of t(), J=6.5Hz), 4.56(1H, t, J=5.3Hz); 5.28(2H, s), 6.88(2H, d, J=8.5Hz) and 7.23(2H, d, J=8.5Hz);

V _max（KBr）3314（m），3250（s），1695（s），1685（s），1634（s），1611（s），1607（s），1578（m），1030（m）and 756（m）cm ^-1;

M/e(CI) 498(MH ⁺, 100%), 35(40), 448(12), 396(10) and 121(7);

Find C, 62.92; H, 7.10; N, 14.22; C ₂₆H ₃₅N ₅O ₅Satisfy C, 62.76; H, 7.09; N, 14.08%.

Embodiment 3

8-[4-acetoxyl-3-(acetoxyl methyl) butyl amino]-1,3-two (cyclopropyl methyl)-heteroxanthine

With the method identical with embodiment 1 from 8-amino-1, it is 8-[4-acetoxyl-3-(acetoxyl methyl of 32% that 3-two (cyclopropyl methyl)-heteroxanthine prepares output) butyl amino]-1,3-two (cyclopropyl methyl)-heteroxanthine, mp163～4 ℃.

δ (CDCl ₃) 0.41-0.49(8H, m), 1.24-1.37(2H, m), 1.72(2H, q, J=6.9Hz), 2.08(6H, s), 2.13(1H, t, J=6.3Hz), 3.59(2H, dt, J=6.9,6.0Hz), 3.68(3H, s), 3.90(4H, the overlapping d of t(), J=7.0Hz), 4.06-4.20(4H, m), 4.50(1H, t, J=6.0Hz);

Find C, 58.05; H, 6.97; N, 14.53; C ₂₃H ₃₃N ₅O ₆Satisfy C, 58.09; H, 6.99; N, 14.73%.

Embodiment 4

1,3-two (cyclopropyl methyl)-8-[4-hydroxyl-3-(methylol) butyl amino)]-heteroxanthine

With the method identical with embodiment 2 from 8-[4-acetoxyl-3-(acetoxyl methyl) butyl amino]-1,3-two (cyclopropyl methyl)-heteroxanthine prepare output be 59% 1,3-two (cyclopropyl methyl)-8-[4-hydroxyl-3-(methylol) butyl amino]-heteroxanthine, mp173 ℃;

δ（CDCl ₃）0.39-0.48（8H，m），1.25-1.39（2H，m），1.69（2H，q，J＝6.6Hz），1.83（1H，m），3.49（2H，m），3.64（3H，s），3.67（4H，t，J＝5.5Hz），3.87（2H，d，J＝7.2Hz），3.91（2H，d，J＝7.2Hz），4.04（2H，t，J＝5.5Hz），6.17（1H，t，J＝5.5Hz）.

Find C, 58.11; H, 7.56; N, 17.98; C ₁₉H ₂₉N ₅O ₄Satisfy C, 58.29; H, 7.47; N, 17.89%.

Embodiment 5

8-[4-acetoxyl-3-(acetoxyl methyl) butyl amino]-1,3-two (cyclopropyl methyl)-7-(methoxymethyl) xanthine

Prepare title compound (98～99 ℃ of mp) with describing identical method with embodiment 1 example.

Pharmacology data

The inhibition of phosphodiesterase

The separation of phosphodiesterase

Prepare Ca from centre chamber ²⁺The PDE(PDE I that/calmodulin is excited is referring to table 1 and Beavo and Reifsynder(1990) nomenclature), then, chromatogram on Mono Q post is by Ca ²⁺The cut and the calmodulin of the active hormesis of expression PDE are compiled, further purifying on calmodulin affinity post.The PDE(PDE II that cGMP-is excited), the PDE(PDE III of cGMP-inhibition) the PDE(PDE IV special) all separate from guinea-pig ventricular with cAMP-.Beginning is isolated the PDE III with chromatography from the peak that contains PDE II and PDE IV on 20ml Mono Q post, the latter's independent stratographic analysis once more on 1ml Mono Q post.On DEAE-Mierocrystalline cellulose and Mono Q post, obtain cGMP-selectivity PDE(PDE V from the pig lung) with chromatography; Remove residual PDE I activity with calmodulin affinity post.

The characteristic of phosphodiesterase isoenzyme

Except the PDE II showed positive interoperability, all preparations were all represented simple Michaelis-Menton kinetics (referring to table 1).

This isozyme activity of PDE I is by Ca ²⁺-calmodulin title complex intensifies, this isozyme energy hydrolysis cAMP and cGMP, and cGMP is preferred Substrate.

This is that the isozyme activity that acts on is intensified by cGMP with cAMP for the PDE II.This isozyme energy hydrolysis cAMP and cGMP, cGMP are preferred Substrates under primary condition.The activity of this isozyme is not subjected to Ca ²⁺The influence of-calmodulin title complex.

This is that the activity of the isozyme of Substrate is suppressed by cGMP with cAMP for the PDE III.This isozyme energy hydrolysis cAMP and cGMP, cAMP is preferred Substrate.The activity of this isozyme is not subjected to Ca ²⁺The influence of-calmodulin title complex.

This isozyme of PDE IV has the affinity high to cAMP, and its hydrolytic action is not suppressed by cGMP.The activity of this isozyme is not subjected to Ca ²⁺The influence of-calmodulin title complex.

This isozyme of PDE V has the affinity high to cGMP, and the activity of this isozyme is not subjected to Ca ²⁺The influence of-calmodulin title complex.

Phosphodiesterase activity is measured

Method (boronate column) with (by people such as Reeves, 1987 describe) boride post of previous record is measured the PDE activity.Enzyme is placed on 50mM Tris, 5mM MgCl ₂, among the PH7.5,, use in 37 ℃ of insulations 4-30 minute down ³(per minute decomposes 4 * 10 to the ring nucleus thuja acid of H mark ⁵) and ¹⁴The Nucleotide 5 of C mark '-(per minute decomposes 3 * 10 to monophosphate ³) the mensuration enzyme.Stop to measure by ebuillition of heated, on the boride post, from substrate material, isolate ³5 of H mark '-the monophosphate product.Reaction mixture is with 0.5mL, 100mM HEPES[N-(2-hydroxyethyl) piperazine-N '-2 ethane sulfonic aicd], 100mM NaCl, PH8.5 dilution, and the boride post of flowing through.Clean pillar fully with identical damping fluid, use again 6ml, 0.25M acetate wash-out 5 '-Nucleotide.By ¹⁴The C-regenerant judges that the withdrawal amount of product is approximately 80%.In used scope of experiment, all measurement result is all linear with soaking time and enzyme concn.

Obtain IC ₅₀The method of value (inhibitor concentration requires to suppress 50% activity) is: the insulation of isozyme, utilize 1 μ M cGMP (not have Ca as the PDE I ²⁺With regulate albumen) Substrate of PDE II and PDE V and with the Substrate of 1 μ M cAMP as PDE III and PDE IV.

Using the scope of inhibitor concentration is 0.1 * IC ₅₀～100X IC ₅₀

Reference:

BEAVO, J.A and D.H.REIFSNYDER, Primary sequence of cyclic nucleotide Phosphodiesterase isozymes and the design of selective inhibitors.Trends.Pharmacol.Sci.11,150-155(1990).

REEVES M.L., B.K.LEIGH and P.J.ENGLAND, The identification of a new cyclic nucleotide Phosphodiesterase activity in human and guinea-Pig cardiac Ventricle.Biochem.J.241,535-541(1987).

Table 1: the dynamic characteristic of phosphodiesterase

Isozyme Km (μ M) Vmax cAMP

cAMP??cGMP??Vmax??cGMP

I. Ca ²⁺/ calmodulin intensify 36 55

II. cGMP-intensify 45 14 1

III. cGMP-suppress 0.5 0.1 5

IV. cAMP-special 2〉n.d

V. cGMP-is special〉1 N.d

A. the enzyme of representing positive interoperability

＞Km＞100μM

N.d can not determine, because a kind of matrix of not hydrolysis of PDE.

The result

Suppress for embodiment number

PDE?VA??PDE?IV

(IC ₅₀μM)

1??0.2??3

2??0.2??9

The formula I compound is in the organism of living, the inhibition effect of the TNF that is produced by the monocyte of human body

Part 1: the determining of sample

The content inspection that utilization is write down below, formula I compound produce the effect of TNF to human body monocyte in the Living Organism.

According to Colotta, people's such as R. method (J.Immunol.132(2); 936＜1984 〉), buffy coat or the platelet removal residue from blood bank separates the also monocyte of purifying human peripheral blood.These monocytes are implanted in the alveolar disk in 24 holes (24-wellmuiti-dishes), its density is that matrix is arranged is 1 * 10 in every hole ⁶Cells/ml.Allow these cell adhesions reach 1 hour, siphon away supernatant liquid afterwards, speed with 10 units per ml adds fresh matrix (the RPMI-1640(Whitaker Biomedical Products that 1ml contains 1% fetus calf serum (fetal colf serum) and penicillin and Sueptomycin, Whitaker, CA).Above-mentioned cell is being with or without under the test compound situation, cultivates 45 minutes at 1nM-10 μ m dosage range (compound solubilization in dimethyl sulfoxide (DMSO)/ethanol so that in the substratum final concentration of ordinary dissolution be 0.5% dimethyl sulfoxide (DMSO)/0.5 ethanol).Add the cytolipin polyose of 100ng/ml in 10ml phosphate buffered saline (PBS) (PBS) (E.Colio 55: B5[LPS] from Sigma chemicals co) then, containing 5%Co ₂Incubator in cultivate down these substratum in 37 ℃ and reach 16～18 hours.Cultivate when finishing, from cell, remove the supernatant liquid of substratum,, use the activity of the supernatant liquid TNF of following radioimmunoassay method 0.05ml then promptly with the centrifugal removal cell debris of speed of per minute 3000 commentaries on classics.

The active radioimmunoassay of II part: TNF

The PH of sample buffer reagent is 7.4, by 0.01M NaPO ₄, 0.15M NaCl, 0.025M EDTA and 0.1% sodiumazide form.By the described improved chloramine-t method of following III part to human recombinant body TNF(rhTNF) carry out iodate, rhTNF be with people's such as chen method [Nature, 330: 581-583＜1987 〉) obtain.

In order to obtain sample (supernatant liquids of 50 μ l substratum) or rhTNF standard substance, the anti-rhTNF(Genzyme of polyclone rabbit, Boston MA) 1/9000 diluent and 8000cpm ¹²⁵I-TNF joins in the buffer reagent of final 400 μ l volumes, and in 4 ℃ of following incubated overnight (18 hours).Common rabbit serum and goat resist-rabbit IgG(Calbiochem) titrated mutually to reach anti--rhTNF precipitation of maximum, then, make the suitable dilution liquid precipitate, wherein diluent is the common rabbit serum of carrier (1/200), goat anti--rabbit IgG(1/4) and the heparin of 25 units, (Calbiochem), each testing tube adds the above-mentioned title complex of 200 μ l, and 4 ℃ of following incubated overnight.Test tube under the 2000rpm condition centrifugal 30 minutes carefully siphons away supernatant liquid, measures the radioactivity of relevant flap with Beckman Gamma 5500 counters.The straight line change curve of logarithmic value Log is as the usefulness of calculating.The concentration of reading TNF the sample from the rhTNF typical curve, promptly straight line is 157～20, the 000Pg/ml scope.

The radioiodination of III part: rhTNF

Carry out iodate with improved chloramine-T method (people such as Frolik, J.Biol chem, 259:10995-11000＜1984 〉) to rhTNF.Briefly, with 15ml 0.5M KPO ₄With the 10ml carrier free ¹²⁵I(100mCi/ml; ICN) dilution is dissolved in the 5mg rhTNF among 5ml, the 20MM Tris PH7.5.Begin reaction, adding concentration is that 100mg/ml(is moisture) the 5ml aliquots containig of chloramine-T solution.After following 2 minutes of the room temperature, insert additional 5ml aliquots containig, add last 5ml chloramine-T after 1.5 minutes again.

This reaction stops to add continuously after 1 minute 50mM Sodium Pyrosulfite, the 120mM potassiumiodide of 100ml and the 1.2mg/ml urea of 200ml of 20ml.Mix these compositions, allow the reaction mixture pre-packed Sephadex G-25 post (PD10 Pharmacia) of flowing through, carry out balance and wash-out with the phosphate buffered saline (PBS) of the PH7.4 that contains 0.25% gelatin.The high radioactivity material that contains cut is washed, and is stored under-20 ℃. ¹²⁵The special active ingredient of I-TNF is the protein of 80-100mCi/mg.With L929 cytotoxin assay method (by Neale, people such as M.L., Eur.J.Can.Chn.Oncol., 25(1): 133～137(1989)) detect the biological activity of iodate TNF, find the unmarked TNF of 80% biologically active.

IV part: measure TNF ELISA

Also can measure the TNF value with improved basic Sandwich ELIST assay method, its method is documented in people such as Winston (current protocols in Molecular, Biology, 11,2,1 pages of people such as Ausubel, Ed.＜1987 〉) John Wiley and Sons.New York.USA).ELISA utilizes the monoclonal anti-human body TNF antibody (stating as follows) of mouse look as the antibody of collecting, and polyclone rabbit Anti-Human body TNF antibody (stating as follows) is as second antibody.In order to detect, adding peroxidase-conjugated goat resists-rabbit antibody (Boehringer Mannheim, Indianopolis, Indiana, USA, Catalog #605222), add a kind of matrix again to be used for peroxidase (the positive phenylenediamine of 1mg/ml) with 0.1% carbamide peroxide.TNF value from the typical curve calculation sample, typical curve is produced by recombinant chou human body TNF, its method for making be documented among the E.coli (derive from Smithkline Beecham Pharmaccuticals, King of Prussia, PA, USA).

V part: preparation Anti-Human body TNF antibody

With recombinant chou human body TNF immunity BALB/C mouse spleen prepare the monoclonal antibody of human body TNF, that used is improve one's methods (the Nature 256:495(1975) of Kohler and Millstein), the full content of this method is hereby incorporated by reference document.By repeating White(NZW to New Zeland) immunity of rabbit prepares polyclone rabbit Anti-Human body TNF antibody.Described rabbit utilizes in Freund ' s auxiliary fully (DIFCO.IL, emulsified recombinant chou human body TNF immunity in USA).

Conclusion

Determine to exist 8-(4-acetoxyl-3-acetoxyl methyl)-1-butyl amino-1,3-two (cyclopropyl methyl)-7-is right-the methoxy-benzyl xanthine, and this shows the IC of the 0.30 μ M that has an appointment in the sample system of TNF product in the organism alive ₅₀

The endotoxin shock of the irritated mouse of D-gal-

This test is documented in people's such as Galanos the method in (Proc, Natl Acad.Sci USA, 76:5939-43(1979)) especially, and its content is hereby incorporated by reference.Briefly, D-gal(D(+) galactosidase) make various mouse allergy, the result causes death by intracellular toxin.Intravenously (i.v) is introduced D-gal(300-500mg/Kg) allow mouse allergy, make lipopolysaccharides (LPS) dosage be reduced to 0.1 μ g.Simply say, give male C ₅₇The intravenous injection of BL/6 mouse has been dissolved in mixing in 0.20-0.25ml pyrogen-free saline at D(+)-gal(Sigma:500mg/Kg) 0.1 μ g LPS, it is from Salmonella typhosa(Difco Laboratories, Detroit, Michigan, USA), described mouse is a 6-12 week size and from Charles Kiver Laboratories(Stone Ridge, New York, USA).The compound of doing test can add at any time at the forward and backward of intravenous injection LPS/D-gal.In this experiment, after the injection LPS, the general 5-6 of animal subject hour death, but discrete was then die between 24-48 hour.

Measure the TNF activity

Measure the TNF value of blood plasma with improved basic sandwich (Sandwich) ELISA method, this method is documented in people's such as Winston Current protocols in Molecular Biology(Pg 11,2,1, people Ed(1987 such as Ausubel) John Wiley and Sons, New York, USA) in, with the monoclonal anti-young mouse TNF(Genzyme that utilizes the hamster, Boston, MA, USA) Elisa is as the antibody of capturing, and is anti-to utilize-young mouse TNF(Genzyme, Boston, MA, multi-clone rabbit USA) is as the antibody of test.From with recombinant chou mouse TNF(Genzyme, Boston, MA, USA) TNF value in the typical curve experiment with computing mouse of Chan Shenging, relevant by the TNF value that ELISA measures with the value of surveying with the L929 biological detection method, the L929 biological detection method is documented in people's such as Ruff J Immunol, 125:1671 1677(1980) in, i.e. the TNF of 70 pico-grams (Pg) among the active corresponding ELISA of 1 unit in the biological assay.The TNF value that ELISA records drops to 25Pg/ml.

In above-mentioned test, active compound has positive reaction in Living Organism, shows ED as the reduction for serum TNF ₅₀Be reduced to about 50mg/Kg oral administration.

Claims

1, the method for a kind of suitable acceptable salt of medicine of a kind of a kind of compound for preparing formula I or its, formula I is:

R wherein ¹And R ²Difference expression (a) part:

-(CH ₂) _m-A (a)

Wherein m represents 0 or integer 1,2 or 3, and A represents a cyclic hydrocarbon group that replaces or do not replace;

R ³Expression hydrogen, replacement or the alkyl that does not replace, or the aralkyl that replaces or do not replace at aryl moiety; With

R ⁴Expression hydrogen, alkyl or alkyl-carbonyl.

The preparation method comprises a formula II compound and a formula III compound reaction, and formula II is:

R wherein ⁵Expression hydroxyl protecting group, L ¹Represent a leavings group; In addition, carry out selectivity step below one or several as required.

(ⅰ) remove protecting group:

(ⅱ) arbitrary radicals R ^1aBe converted into R ¹And/or arbitrary radicals R ^2aBe converted into R ²And/or arbitrary radicals R ^3aBe converted into R ³

(ⅲ) compound of formula I is converted into another compound of formula I;

(ⅳ) compound of formula I is changed into the acceptable salt of its medicine.

2, a kind of method according to a kind of compound of claim 1 preparation, wherein A represents a cyclopropyl.

3, a kind of method, wherein R according to claim 1 or a kind of compound of 2 preparations ³Represent an aralkyl.

4, a kind of according to each prepares a kind of method of compound, wherein R among the claim 1-3 ³The benzyl that expression replaces or do not replace.

5, a kind of according among the claim 1-4 each prepares a kind of method of compound, wherein R ³Expression 4-methoxy-benzyl.

6, a kind of according to each prepares a kind of method of compound, wherein R among the claim 1-5 ⁴Expression hydrogen.

7, a kind of according among the claim 1-5 each prepares a kind of method of compound, wherein R ⁴The expression alkyl-carbonyl.

8, a kind of according to a kind of method that is selected from any compound among the embodiment 1-4 in the present invention of claim 1 preparation.

9, a kind of method for preparing a kind of pharmaceutical composition, said composition comprises compound or the acceptable salt of its suitable medicine and/or the acceptable solvate of medicine and the medicine acceptable carrier of a formula I, and its method comprises compound or the acceptable salt of its suitable medicine and/or acceptable solvate of its medicine and the medicine acceptable carrier that mixes formula I.