CN1221609A - Medicinal use of adenosine phosphates and their pharmaceutical preparations - Google Patents

Abstract

The application of 2-chlorodeoxyadenosine phosphate, 2-chloro-2 '-arabinose-fluoro-2' -deoxyadenosine phosphate or their medicinal salt in preparing medicine for treating mammal skin cell hyperproliferation disease and the prepared medicinal preparation.

Description

Medicinal and the pharmaceutical preparation of adenylic acid esters

The application is dividing an application of applicant's PCT application of submitting, entering on January 26th, 1998 the China national stage on June 6th, 1996, original applying number 96 1 95944.4, denomination of invention are " nucleotide analog that is used for topical treatment of proliferative disease of skin ".

The invention provides and be used for the preparation that the topical therapeutic psoriasis reaches other disease that is excessively caused by the Skin Cell hypertrophy.Particularly, the present invention relates to utilize preparation for treating psoriasis and other dermatosis of the phosphate ester that contains nucleoside analog, anti-hypertrophy nucleoside and the end capped di-deoxynucleoside of DNA chain.

Psoriasis is the application on human skin benign disease, and its feature is that generally the squama that speckle is thickened covers.This disease is that the epidermal cell proliferation increase by unknown cause causes.In normal skin, cell moves to the required time of top granular layer from bottom and was approximately for five weeks; And in psoriasis, part is because the increasing and the increase of somatoblast ratio of proliferative cell quantity, and this time only is 6 to 9 days (G.Grove, Int J.Demato1.18:111,1979).Nearly 2% people suffers from psoriasis in the U.S. population, wherein nearly 3% Caucasia American about 1% African American, and seldom the native country American suffers from this disease.

With the epidermal hyperplasia of inflammation-related be the feature of psoriasis skin.The Skin biopsy of infected zone demonstrates the hyperkeratosis that keeps with nuclear fragmentation, along with the hypertrophy increase and the chronic inflammatory disease of defectiveness keratinization are soaked into.(referring to EA Bauer, M Tabas and J.B.Goslen, internal medicine textbook (TexbookofInternalMedicine), W.N.Kelly (editor), 1989, pp1042-1045).Along with hyperplasia increases, DNA is synthetic in the infected tissue increases, and described infected tissue has been the basis that is used to assess the efficacy determinations of antipsoriatic.

Although reported the rare primates (N.J.Lowe, DrugDev.Res.13:147-155,1988) with the clinical and histopathology feature of psoriasis, that does not generally acknowledge is used for psoriasic animal model.The research of therefore treating the psoriasis medicine depends on the animal of the spontaneous mutation (fsn) that stands easily to peel off skin or the experiment in the mice bacterial strain and induces hypertrophy people such as (, J.InvestDermatol92:414,1989) J.P.Sundberg.Be used for another mouse model with the epidermal hyperplasia diseases associated and be must FAP (EFAD) hairless mouse.

Test inductive animal model and also comprise athymism nude mice, this mice has immunologic defect and transplants the application on human skin that disease is arranged.

Current therapy is made up of the trial that reduces rapid hypertrophy of cell and minimizing inflammation.These therapys comprise partly, systematically use activating agent, perhaps both, this therapy can be at random in conjunction with irradiation.Topical therapeutic comprises use steroid cream and (UV B 290-320nm) uses coal-tar ointment afterwards at ultraviolet radiation.People have obtained some successes on 5-fluorouracil in the part, but this treatment can cause and forms serious erythema, edema, bulla and at the area for treatment skin ulcer, therefore can not be accepted (C.J.McDonald, Pharmac.Ther.14:1-24,1981) by the patient well.Once Triazure is crossed in local test on psoriatic's skin ^TM(6-aza uridine triacetate), but effect (William Drell, person-to-person communication (personal commuication), in May, 1993) do not had.

The anti-proliferative agent that comprises methotrexate, 6-aza uridine and 6-azauridine triacetate was also once systematically used.Large-area psoriasis is by oral; 8-methoxypsoralen-a kind of photosensitizer then carries out ultraviolet light,long wave (320nm) and shines or take retinoid such as etretinate, then carries out ultraviolet light,long wave irradiation treatment (C.J.McDonald, Pharmac.Ther.14:1-24,1981; E.A.Bauer, M.Tabas and J.B.Goslen, in Textbook of Internal Medicine, W.N.Kelly (ed.), 1989, pp1042-1045).Other disease that is caused by the Skin Cell hypertrophy comprises atopic dermatitis, lichen planus, actinic keratosis, basal cell carcinoma and squamous cell carcinoma.Other disease that the topical use of the phosphate ester of anti-hypertrophy nucleoside and di-deoxynucleoside and analog thereof also is not in the news and is used for the treatment of psoriasis or is caused by the Skin Cell hypertrophy.

Phosphate ester such as acyclovir that the open WO 94/26273 of relevant PCT discloses the herpes nucleoside are effective in the animal by the mutated viruses strain infection, wherein owing to sudden change infective virus enzyme, thymidine kinase, acyclovir can not be transformed into the acyclovir phosphate ester.

The method of other disease that the invention provides the treatment psoriasis and caused by the Skin Cell hypertrophy, this method comprise that topical application contains nucleoside analog phosphate ester with following formula or its officinal salt as composition of active components: Wherein N is the nucleoside analog with anti-proliferative activity; Z is O or S or NH; And n is 1,2 or 3.This method can contain nucleoside analog phosphate ester or its officinal salt with following formula by use and carry out as composition of active components:

Wherein N is nucleoside or the nucleoside analog with anti-proliferative activity; X is O or CH ₂Or S; Z is O or S or NH; And n is 2.

In preferred embodiments, nucleoside or nucleoside analog be selected from cytosine arabinoside, arabinose guanosine (guanosine arabinoside), floxuridine, 5-floxuridine, 6-aza uridine, 2-chlorodeoxyadenosine, 2-chloro-2 '-fluorine arabinose deoxyadenosine, 2-fluorine arabinose deoxyadenosine, 6-methyl mercapto purine nucleosides, zalcitabine, Didansine, dideoxy guanosine, didanosine, DIDEOXYADENOSINE, 2 '-deoxidation tubercidin, 2 '-deoxidation homotype mycin, 2 '-deoxidation-(3,4-d) pyrimidine, acyclovir and ganciclovir.

The method according to this invention, the concentration of active component is 0.001gm% to 100gm% in the compositions; In preferred embodiments, the concentration of active component is 0.01gm% to 10gm% in the compositions; In particularly preferred embodiments, the concentration of active component is 0.1gm% to 5gm% in the compositions.

According to an aspect of the present invention, in skin area every day set of applications compound 1 to 10 time, each application dose is 0.001gm% to 100gm%.Another kind method be in skin area every day set of applications compound 1 to 10 time, each application dose is 0.01gm% to 10gm%.In preferred embodiments, in skin area every day set of applications compound 1 to 10 time, each application dose is 0.1gm% to 5gm%.

Another aspect of the present invention provides a kind of pharmaceutical composition, and said composition comprises nucleoside analog phosphate ester in the described method as active component.In preferred embodiments, described pharmaceutical composition also contains propylene glycol, PEG400 and Polyethylene Glycol 3350.In particularly preferred embodiments, described compositions also contains the composition that can strengthen dermal osmosis in topical application.

The present invention also provides the phosphate ester of the anti-hypertrophy nucleoside analog that is used for the inventive method.In an embodiment preferred, nucleoside analog is phosphoramidate or thiophosphate and/or methylene phosphate ester or Thiophosphonate.

The invention provides the phosphate ester of anti-hypertrophy nucleoside, as 2-chlorodeoxyadenosine, 2-chloro-2 '-fluorine arabinose deoxyadenosine, cytidine or guanosine arabinose glycosides, 6-aza uridine, 6-MPR, 5-floxuridine, floxuridine, acyclovir, ganciclovir, zalcitabine and other di-deoxynucleoside and the end capped nucleoside analog of chain; When they during by topical application, its reduce hyperplasia speed and eliminate on lymphocyte in the psoriasis skin wondrous and unusual ground effective.

Monophosphate, bisphosphate and triguaiacyl phosphate have following general formula: Wherein N is anti-hypertrophy nucleoside analog; Z is O, S or NH; And n is 1 or 2; Perhaps they have following formula:

Wherein N is anti-hypertrophy nucleoside analog; Z is O, S or NH; X is O, CH ₂Or S; And n is 1,2 or 3.

Therefore, described phosphate ester can be phosphate ester, thiophosphate (phosphothiorate) or phosphoramidate, and diester and three esters can have the bridge atom except that oxygen atom, as 2, and 3-μ-thio triphosphates ester or 2,3-μ-Medronate ester.

Opposite with expected result, these nucleoside analog phosphate esters can reduce cell division speed astoundingly by the cell membrane of the excessive Skin Cell of hypertrophy and by the various enzyme steps in the biosynthesis that suppresses purine and pyrimidine, nucleotide, RNA and DNA.In addition, chemical compound 2-chlorodeoxyadenosine phosphate ester and 2-chloro-2 '-fluorine arabinose deoxyadenosine phosphate ester is specially adapted to treat psoriasis inflammation part, and this is because its inhibition is present in the lymphocyte in psoriatic's scytitis infiltrate and the ability of monokaryon leukocyte growth.The present invention also provide nucleoside analog one-, two-and the pharmaceutical preparation of triguaiacyl phosphate, the concentration of wherein said phosphate ester should reduce the hypertrophy of psoriasis Skin Cell effectively when topical application.When being used for skin with suitable topical formulations, the end capped di-deoxynucleoside phosphate ester of DNA chain can reduce the psoriasis proliferation of cells similarly.They comprise acyclovir, ganciclovir, zalcitabine, Didansine, dideoxy guanosine, DIDEOXYADENOSINE, didanosine, 3 '-nitrine Didansine (AZT) and other di-deoxynucleoside analog, as unsettled U.S. Patent application SN 07/373, described in 088 those, the document is hereby incorporated by.

The other nucleotide phosphate that can be used as antipsoriatic when use the part comprises nucleotide metabolism (Nucleotide Metabolism) (J.F.Henderson; A.R.P.Paterson, AcademicPress, 1973, those that enumerated in pp266-271).

The salt of these chemical compounds can easily make, and this class salt must demonstrate very strong dissolubility in aqueous medium (as cream, colloid or other aqueous dispersion).The typical useful salt of these chemical compounds comprises sodium salt, potassium salt, lithium salts, ammonium salt or hydrogen salt.Also can use any acceptable cation of physiology that well known to a person skilled in the art.And this class salt is useful and effective in that the good mucosa or the Polyethylene Glycol unguentum and the washing liquid of skin decentralized photo are provided.

Topical compositions

Another aspect of the present invention provides and has been used for the psoriasic method of topical therapeutic, and this method comprises that the compositions that will contain nucleoside analog phosphatidic acid of the present invention is applied to psoriasis infringement place of infected patient skin.

The nucleoside analog phosphate ester of the present invention of foregoing topical application, di-deoxynucleoside and the end capped nucleotide phosphate of chain can make by introducing the various compositionss that well known to a person skilled in the art, described known compositions is meant useful and compositions easily in the dermatological purposes.Nucleotide phosphate is more water-soluble than corresponding alkali usually, and therefore, aqueous solution, water-in-oil emulsion or water cream are very preferred preparations.The water solublity of nucleoside analog phosphate ester of the present invention can be improved by preparing its salt such as sodium salt, potassium salt, lithium salts, ammonium salt or hydrogen salt.In particularly preferred compositions, active component makes in Polyethylene Glycol (PEG) vehicle.In addition, by utilizing insoluble powder such as starch or Pulvis Talci as solvent or diluent or carrier, can be with dry powder formulations form topical application of active composition.

Because vehicle may be selected to and can improve permeability, prolongs active duration or satisfy the requirement of medication position, so vehicle is the important component in some topical compositions.For example, the preparation that is applied to health callus position such as palm or sole can comprise and improves penetrating agent such as dimethyl sulfoxide, propylene glycol or Azone ^TMIn addition, powder composition may be selected to and is applicable to that the intertrigo zone is as between human body two lower limb crotches, interior elbow or finger or the toe.Compositions also can be made into and contains various organic polymers or well known to a person skilled in the art other compositions, thereby obtains the lasting release that activity is controlled the psoriasis derivant.

A large amount of topical compositions that are fit to can find in the known formulary of Pharmaceutical Chemist: Blaug, S., Ch.87 in Remington ' s Pharmaceutical Sciences (15th Ed., 1975, Mack Publishing Company, Easton, Pennsylvania 18042).These compositionss comprise as powder, paste, ointment, gel, wax, oil, lipoid, no water absorbent, oil-in-water or water in oil emulsion, carbowax Emulsion (different molecular weight polyethylene glycol), semi-solid gel and the semi-solid mixtures that contains carbowax.

The concentration of active component can be about 0.001gm% to 100gm% in the topical compositions of the present invention, preferably about 0.01gm% to 10gm%, and most preferably from about 0.1gm% is to about 5gm%.Said composition can further comprise other reagent that helps lend some impetus to dermal osmosis and healing of valid density, as above-mentioned with reference to describe in the formulary and the high those of ordinary skill in this area known those.

The effectiveness that contains the topical compositions of the active nucleoside analog phosphate ester of the present invention can be used and well known to a person skilled in the art that the routine test method assesses.These comprise using predicts mensuration (N.J.Lowe, Drug Dev.Res.13:147-155,1988) in simulated animal model (as epidermal hyperplasia) and other body.

But the compositions repeated application is in psoriasis infringement place of skin infection, as once a day, twice or several times; Treatment can continue a couple of days, until reaching healing.Toxicity takes place in it and danger hypersensitive is minimum.

In particularly preferred embodiment of the present invention, nucleoside by 2-chlorodeoxyadenosine, 2-chloro-2 '-fluorine vidarabine, 6-aza uridine, 5-floxuridine or floxuridine, guanosine arabinose glycosides or di-deoxynucleoside and local unguentum form, described local unguentum is made up of the mixture that contains Polyethylene Glycol 3350, PEG400 and propylene glycol or ethylene glycol.

Nucleoside analog is made up of following material: cytidine or guanosine arabinose glycosides, floxuridine, 5-floxuridine, 6-aza uridine and DNA chain end-blocking di-deoxynucleoside, as zalcitabine, Didansine, dideoxy guanosine, didanosine, and the di-deoxynucleoside analog, as the chemical compound described among the open WO 90/00555 of PCT and related compound 2-chlorodeoxyadenosine and 2-chloro-2 '-arabinose fluorine deoxyadenosine and 2-fluorine arabinose deoxyadenosine.When with one-, two-or triguaiacyl phosphate when transmitting, other nucleoside analog that can suppress epidermal dna polymer enzyme comprises 2 '-deoxidation tubercidin, 2 '-deoxyformycin and 2 '-deoxidation-(3,4-d) pyrimidine.

Can be used as local give additional compounds with antipsoriatic comprise one of following ribonucleotide and dezyribonucleoside derivant-, two-and triguaiacyl phosphate: guanozola, Ismipur, 6-thio-purine, 6-methyl mercapto purine, 2,6-diaminopurine, 8-azepine xanthine, formycin, psicofuranine, decoyinine A, xyloyl adenosine, 6-chloropurine, 6-azauracil, 5-fluorine cytidine or 5-flurocytosine, 5-fluorouracil, 5-iodouracil and 5-bromouracil; And comprise 3 '-(chemical constitution can be referring to nucleotide metabolism, J.F.Henderson for deoxyadenosine, deoxidation xylopyranose base thymidine, deoxy-glucose pyrans glycosyl thymidine and cytosine arabinoside; A.R.P.Paterson, Academic Press, 1973, pp.266-271) one-, two-and triguaiacyl phosphate.

The mensuration of topical drug's assessment

1. synthetic inhibition of epidermis DNA measured.Because the DNA that increases is synthetic is to be feature with the epidermal hyperplasia in the psoriasis skin, has been used to assess the synthetic inhibition degree of the DNA that is caused by medicament so mix radioactivity DNA precursor in the skin histology that utilizes pharmaceutical treatment.In brief, the antipsoriatic in the topical vehicle is applied in the athymic mouse zone, wherein utilizes technology known in the art will suffer from psoriasic human dermis and epidermic grafting to described zone.Similarly, EFAD hairless mouse model can be used in the mensuration, thereby record the effect of anti-proliferative drug.Greatly after treatment 6 hours, the excision tissue for the treatment of is placed on and contains ³H-thymidine deoxyribose or ³Carry out 0.1-5 hour pulse labeling in the tissue culture medium (TCM) of H-bromodeoxyribouridine.Utilize the conventional method DNA isolation then, be incorporated among the DNA by radioactivity (the cpm/ μ gDNA) measurement of measuring in institute's DNA isolation ³H-thymidine deoxyribose or ³H-bromodeoxyribouridine amount.Be incorporated into the DNA radioactivity of radioactivity and the compositions of utilizing different antipsoriatics (% of active component increases) among the DNA by treated tissue more never, can measure antipsoriatic and suppress outgrowth effectiveness.

2. the polyamines biosynthesis suppresses to measure.This mensuration is based on the amount of the ODC Ornithine decarboxylase that exists in the epidermis cell (ODC).ODC is the enzyme of limiting speed in polyamines forms, and described polyamines increases in hypertrophy usually, and described hypertrophy comprises the epidermal hyperplasia relevant with psoriasis.(people such as Lesiewicz, Models in Dermatology, vol.2, the pp112-116 (H.I.Maibach﹠amp of stripping off by the horny layer band; N.J.Lowe, eds.), 1985), or, also can cause the active increase of ODC by using phorbol ester (12-O-four capryl phorbol-13-acetas or TPA).

3. Skin biopsy.The athymic mouse that in 1-15 days, utilizes antipsoriatic thing treatment EFAD hairless mouse, fsn/fsn easily to peel off the skin mice or transplant with the psoriasic application on human skin of trouble, the skin that inspection is treated, thereby obtain psoriasic thick indication (redness of the peeling off of skin, inflammation etc.), utilize biopsy to analyze then, utilize the quantity of normal structure method count enable cuticular cellulose at microscopically.With the quantity of cuticular cellulose with similarly normally do not treat psoriasis skin and compare, thereby determine to utilize the effect of antipsoriatic thing treatment.Moreover, by measure in the epidermal hair tubule leukocyte (being neutrophilic leukocyte and/or monocyte) quantity and with respect to the telangiectasis of not treating psoriasis skin and normal control skin, the inflammation relative extent in can quantitative tissue.

Similarly, utilize people's clinical trial of Skin biopsy and microscopy to be undertaken by the both sides paired comparison that use has an antipsoriatic agent that is applied to about 3cm diameter skin area (on forearm or lower limb) on a small quantity.Preferred especially both sides paired comparison is to avoid by the danger of area for treatment to the cross action of non-treatment control zone.

4. use and easily peel off the skin mice.The inbred mice that to stand easily to peel off the spontaneous mutation (fsn/fsn) of skin develops into white flaky skin, and after wean, this flaky skin thickens with advancing age and constantly, forms hypertrophy dermatosis, particularly psoriasis thus.During to age of 42 days, mice demonstrates the epidermal hyperplasia that has multilamellar counterdie and inflammation, and this measures by leukocyte and crooked dermovascular increase, and all these features all are the psoriasic features of people.These mices can be used as people's psoriasis model that the treatment of determining to utilize the topical application nucleotide phosphate is renderd a service.To comprise that the compositions of nucleotide phosphate is applied to the 1-5mm part of dorsal part or the veutro skin of fsn/fsn mice, and to afterwards 15 days, observed skin from 12 hours roughly and microscopically, thereby utilize conventional dermatological and Histological method to determine the degree of epidermal hyperplasia and other symptom of the above-mentioned psoriasis skin that occurs.Carry out relevant comparison with the similar portions of untreated mice, compare with other untreated fsn/fsn mice, only contain carrier components and not have the fsn/fsn mice of the combination treatment of nucleotide phosphate to compare with utilizing, and to compare with the not mutated mice of the similar treatment in fsn site.

5. keratinocyte labelling.It is consistent that the variation of protein expression changes with the hypertrophy relevant with psoriasis skin.Two kinds of molecules that are used for the detection of skin abnormal diseases are keratin and tenuin (filaggrin).The gathering of special mice keratin K6 in the base portion upper epidermis is that general relevant with hypertrophy molecule changes.In not having hair EFAD mice or fsn/fsn mice, can be used for determining the molecule that occurred in the base portion upper epidermis and the amount of this molecule based on the method for detection of antibodies K6, therefore can be used as the labelling that treatment psoriasis medicine is renderd a service.

Tenuin is a kind of protein in the epidermis, and it is relevant with the keratohyalin granule in granular layer and the transition cell usually, but this tenuin disappears in the keratinocyte of top.Yet in the fsn/fsn mice, tenuin still remains resident in the superficial cell layer with the aspect of model of transition cell.As by anti-tenuin antibody test and/or by microscopic observation, having tenuin in the keratinocyte of top is the feature of psoriasis cell, can be used for the effectiveness of evaluate application in the treatment psoriasis medicine of fsn/fsn mouse skin.

Synthetic purine and the purine nucleoside analogs of selecting.This paper has summarized the concrete grammar of more synthetic anti-hypertrophy purine and purine nucleoside analogs, and described purine and purine nucleoside analogs can be used as the psoriasic phosphate ester of treatment.Additional compounds and synthetic method are known in those skilled in the art.Representational method is disclosed in RK.Robins and G.D.Kini, antineoplastic agent chemistry (TheChemistry of Antitumour Agents) (D.EV.Wilman volume), and 1990, among the p299-321.

(1) purine analogue

At high temperature, in 1,2,3,4-tetralin, utilize Phosphoric sulfide to handle hypoxanthine, synthetic Ismipur.

In the backflow pyridine, utilize Phosphoric sulfide to handle guanine, can synthesize 6-thioguanine.

By 3 of the suitable replacement of closed loop in imidazoles [4,5-c] pyridine ring system, 4-di-amino-pyrimidine, synthetic 3-denitrogenation adenosine, 3-denitrogenation hypoxanthine and 3-denitrogenation-Ismipur (Robbins, people such as R.K., J.Org.Chem.28:3041,1963).

When utilizing phosphorus oxychloride to make 5 (4)-carbamoyl Methylimidazole .s-4 (5)-carboxylate methyl ester dehydration, preparation can be synthesized the 3-deazaguanine when 5 (4)-cyano methyl imidazoles-4 (5)-carboxylate methyl ester of Liquid Ammonia Treatment cyclisation.

(2) purine nucleosides

Utilize catalyst mercuric chloride, condensation 6-methyl mercapto purine and 2,3,5-three-O-acetyl group-D-ribose-furyl glycosyl chlorine then carries out deacetylation, can synthesize 6-methyl mercapto purine nucleosides.In addition, 6-methyl mercapto purine nucleosides can directly make (Fox, people such as J.J., J.Am.Chem.Soc.80:1669,1958) by the 6-NSC-40774 that methylates.

9-β-D-arabinofuranosyl adenin glycosyl adenine (ara-A) can be synthetic by the following step: utilize mesyl chloride handle 3 of 9-(β-D-wood furyl glycosyl) adenine '; 5 '-two-O-isopropylidene derivant; that it is transformed into is corresponding 2 '-O-mesyl derivant; by acetic acid this derivant is cracked into 9-(2 '-O-mesyl-β-D-wood furyl glycosyl) adenine then; in methanol, handle with Feldalat NM again and make 2 '; 3 '-epoxide; in moisture dimethyl formamide, handle open loop with sodium; generate ara-A, the recrystallize purification.

Related compound 9-β-D-arabinofuranosyl adenin glycosyl-2-fluoroadenine (2-fluoro-ara-A) can be synthetic by the following step: in the backflow pyridine, utilize acetic anhydride acetylation 2, the 6-diaminopurine, generate 2,6-diacetylamino purine is then with 2,3,5-three-O-benzyl-alpha-D-arabinofuranosyl adenin glycosyl chlorine condensation generates block β-anomer nucleoside; In ethanol, make its deacetylation by utilizing methylamine to handle; generate the diaminourea nucleoside; utilization contains the fluoboric acid and the tetrahydrofuran compound of sodium nitrate aqueous solution and handles; make its generate 2 '; 3 '; 5 '-three-O-benzyl-β-D-arabinofuranosyl adenin glycosyl-2-fluoroadenine utilizes boron chloride to handle, and generates 2-fluoro-ara-A.

By condensing 2,6-dichloropurine and 1,3,5-three-O-acetyl group-2-deoxidation-D-is red-furan pentose and by other method (Robins and Kini, the antineoplastic agent chemistry, 1990, p308), but Synthetic 2-chloro-2 '-deoxyadenosine.

(3) purine ring nucleotide and cyclic nucleotide analog

8-chloro-cAMP can be synthetic by the following step: in the presence of sodium hydrate aqueous solution, the cAMP bromination is become 8-bromo-cAMP, at high temperature utilize thiourea that it is processed into 8-sulfo--cAMP; In the HCl methanol solution, utilize chlorinated with chlorine.

(4) other anti-hypertrophy nucleoside relevant with purine

Tiazofurine (2-β-D-ribose furyl glycosyl)-thiazole-4-carboxamide can be synthetic by the following step: utilize ethyl bromide acetone to handle 2,5-dehydration-3,4,6-three-O-benzoyl-D-allose base thioamides, promote closed loop, generate 2-(2,3,5-three-O-benzoyl-β-D-ribose furyl glycosyl) thiazole-4-carboxylic acid ethyl ester is major product; Separate then and handle with methanol ammonia.

Related compound seleno tiazofurine can be synthetic by the following step: utilize liquid H ₂Se handles 2,5-dehydration-3,4, and 6-three-O-benzoyl-D-allose nitrile generates corresponding selenonyl amine; With the ethyl bromide acetone reaction, promote closed loop, generate 2-(2,3,5-three-O-benzoyl-β-D-ribose furyl glycosyl) selenazoles-4-carboxylic acid, ethyl ester, be major product; Separate then and handle with methanol ammonia.

.beta.-Pyrazofurin (3-(1-β-D-ribose furyl glycosyl)-4-hydroxypyrazoles-5-Methanamide) is naturally occurring nucleoside, can separate (Gutowski, people such as G.E., Biochem.Biophys.Res.Cornmun.51:312,1973) from streptomyces candidus.In addition .beta.-Pyrazofurin can followingly synthesize: utilize hydrazine derivate to handle α-ketone ester, generate two azo intermediate; With acetic anhydride and sodium acetate heating, generate cyclisation hydroxypyrazoles nucleoside; Use the methanol ammonia amination, debenzylation generates .beta.-Pyrazofurin then.

2-deoxycoformycin (8R)-3-(2-deoxidation-β-D-red-furan pentose base)-3,6,7,8-imidazolidine [4,5-d] [1,3]-diaza -8-alcohol can be synthetic by the following step: with aglycone, 6,7-glyoxalidine [4,5-d] [1,3]-and diaza -8 (3H)-ketone and 2-deoxidation-3,5-two-O-is right-toluoyl-D-is red-and furan pentose base chlorine carries out glycosylation (Chan, people such as E, J.Org.Chem.47:3457,1982).

The similar thing of synthetic nucleosides phosplate, bisphosphate and triguaiacyl phosphate and nucleotide phosphate: by nucleoside and triclosan oxidation phosphorus reaction and the method for synthetic nucleosides phosplate is described in (people such as Yoshikawa in open WO90/00555 and WO 94/26273 and the above-described document, Bull.Chem.Soc.Japan 42:3505-3508,1969; Toorchen, D. and Topal, M, Carcinogenesis 4:1591-1597,1983).The nucleoside diphosphate ester can be according to Ott, people's such as D.G. method preparation (Anal.Biochem.21:469-472,1967).

The ribonucleoside triphosphote ester can be prepared by following method: the method for Seela and Roling (Nuc.Acids Res.20:55-61,1992); Or the method (J.Am.Chem.Soc.83:663,1991) by Moffat and Khorana, make by single-nucleotide phosphate; Or the method for Hord and Ott (J.Am.Chem.Soc.87:1785-1788,1963).The following example 1-3 has described some detailed synthetic methods of the phosphate ester that is used to prepare nucleoside and nucleoside analog.

The similar thing of other nucleotide phosphate, comprise nucleoside thiophosphate, nucleoside phosphoramidate, nucleoside phosphate ester and nucleoside fluoro phosphate ester, can be synthetic by well known to a person skilled in the art method, these methods by the D.W.Hutchinson summary (the nucleoside list-, two-, three-and synthetic, the reaction of four phosphate esters and nucleotide and the variation of character and phosphinylidyne residue.Nucleotide and nucleoside chemistry (Chemistry of Nucleotides and Nucleosides), L.Townsend, ed., 1991, pp81-146 and the list of references of being quoted thereof).Common synthetic method is summarized as follows.

(1) the nucleoside thiophosphate is a nucleotide analog, and wherein one or more phosphoryl oxygen atoms are replaced by sulfur.Early stage synthetic method is with protected nucleoside and three (1-imidazole radicals) phosphine (phosphane) reaction of Salmon-Saxl, and nearest synthetic method is to utilize thiophosphoryl chloride (PSCl ₃) replacement back one reagent.By handling with sodium hydroxide and Carbon bisulfide, nucleoside aniline phosphate ester (phosphoranilidate) can be transformed into thiophosphate.Nucleoside 5 '-thiophosphate can by nucleoside 5 '-the direct sulfuration of phosphite ester obtains.Purine nucleosides 2 ' (3 ')-thiophosphate can be synthetic by the following step: with 2 ', 3 '-O-di-n-butyl stannylene derivant is reacted with thiophosphoryl chloride, and then alkaline hydrolysis.

(2) nucleoside phosphoramidate is a kind of analog, and wherein one or more phosphoryl oxygen atoms are replaced by nitrogen, forms the P-N key; Even under the moderate acid condition, this P-N key is more unstable more than the P-S key of nucleoside thiophosphate.Synthetic these chemical compounds comprise the phosphorylation of aminonucleoside, and utilize phosphotriester to handle the nucleoside azide.The oleophylic nucleoside phosphoramidate is useful especially treatment psoriasis chemical compound, and reason is that they are absorbed easilier by cell, is hydrolyzed into bioactive compound then.

(3) nucleoside phosphate ester is a kind of chemical compound, and wherein the oxygen atom of phosphoryl is replaced by carbon, forms stable P-C key; When replacing phosphorus atoms by the alkyl group of giving electronics that replaces oxygen, this chemical compound has the acidity of the P-OH group of reduction.For those skilled in the art, nucleotide phosphate utilization easily or Arbusov or Michaelis-Becker reaction are made by nucleoside halogenide.Nucleoside 5 '-phosphonate ester can utilize method well known to those skilled in the art by 2 ', 3 '-protected 5 '-iodo-5 ' deoxynucleoside is synthetic.Deng join nucleoside 5 '-phosphonate ester (wherein 5 '-oxygen replaced by methylene group) can synthesize through the following steps: the nucleoside 5 of coupling due care '-aldehyde and the inferior phosphoranyl methylphosphonic acid of triphenyl diphenyl; obtain α; β-unsaturated phosphonic acid diester; and then the reduction and on the phosphoryl residue deprotection, obtain phosphonate ester.Deng join nucleoside 3 '-phosphonate ester can synthesize through the following steps: begin by phosphono ribose-1 chloride, with the heavy metallic salt coupling of this chemical compound and purine or pyrimidine.Therefore the polarity of phosphonate ester is usually less than its phosphate ester counter pair, can be used as antipsoriatic, and they were absorbed easilier by cell when reason was topical application.

(4) nucleoside fluoro phosphate ester is the similar thing of a kind of mononucleotide.By 2 of nucleotide, 4-dinitrophenyl ester utilizes 2, the 4-dinitrofluorobenzene handle nucleoside 5 '-phosphate ester make nucleoside 5 '-the fluoro phosphate ester.

(5) the similar thing of other nucleotide phosphate comprises that those atoms except that oxygen atom are at nucleoside two-and the α of triguaiacyl phosphate, between β-phosphorus atoms or at the β of ribonucleoside triphosphote ester, substituted phosphate ester (is included in D.W.Hutchinson between γ-phosphorus atoms, nucleotide and nucleoside chemistry, LTownsend, ed., listed chemical compound in the 119th of 1991 the page the table III).Normally, α, β-analog can prepare through the following steps: by means of DCC condensation 2 ', 3 '-protected nucleoside of O-and pyrophosphate analog; Perhaps by nucleophilic displacement reaction, this reaction comprise utilize 5 of methylene biphosphonic acid esters ion substitution tosyl yl nucleosides '-tosyl (tosyl) residue on the saccharide residue of position.The preparation of beta, gamma-analog is shown in embodiment 4.

Local preparation with polyethylene cream: under agitation, in 70 ℃ with 5 gram nucleotide phosphates be dissolved in 5ml propylene glycol and 60 gram PEG400s (two kinds of reagent are all available from Spectrum ChemicalsInc.Los Angeles, CA) in.Under agitation add 40 gram Polyethylene Glycol 3350 (SpectrumChemicals Inc.), become limpid fully until mixture.Then compositions is placed pipe or other container, be cooled to room temperature and sealing.The valid density of active component can change between 0.001gm% to 50gm% in the compositions, is preferably 0.01gm% to 5gm%.Known as those of ordinary skills, said composition also can contain the composition of other composition (as dimethyl sulfoxide or DMSO) that strengthens percutaneous permeability and/or enhancing composition stability between the storage life.

Psoriasis treatment: the topical compositions of the present invention that contains treatment hypertrophy or antiphlogistic nucleotide phosphate that will treat psoriasis valid density and be 0.01gm% to 10gm% is applied to the infected zone, every day 1 to 6 time.The concentration of the active nucleoside phosphate compound of dosage standard and the best can be easily definite by the experienced doctor in treatment psoriasis field.

Embodiment 1: prepare the ribonucleoside triphosphote ester by the monokaryon glycosides

5 of deoxyribonucleotide, dideoxy ribonucleotide and analog '-preparation of triguaiacyl phosphate comprises the series reaction of following statement.

The synthetic unsettled U.S. Patent Application Serial .08/060 jointly that is described in of mononucleotide is in 258 (applications on May 12nd, 1993).At room temperature, nucleotide (I) and excessive 1,1 '-about 1 hour of carbonyl dimidazoles (II) reaction, form imidazate (III) (imidazolidate).Adding excessive inorganic pyrophosphate (IV) before, make with methanol unreacted 1,1 '-carbonylic imidazole decomposes.This has eliminated the formation of inorganic pyrophosphate, and this pyrophosphate will be removed from reaction material subsequently.After adding inorganic pyrophosphate (IV), finish phosphorus acylation reaction in about 24 hours, utilize the cellulosic anion-exchange chromatography purification of DEAE-ribonucleoside triphosphote ester (V) product then, then product is changed salify such as sodium salt.Because can reacting together, nucleotide (I) and imidazate (III) form symmetric pyrophosphate by-product, so the cellulosic anion-exchange chromatography of DEAE-should carry out under the condition of lower pH, wherein required product (V) has than the electric charge that need not less side products, can separate this two kinds of chemical compounds like this.

Being used for synthetic a kind of reagent is the tributyl ammonium pyrophosphate, and this pyrophosphate can be prepared by the following step: to the pyrophosphoric acid pyridine solution (by (446ml is 1mmol) through Dowex 50W-X4 with tetrasodium pyrophosphate decahydrate solution ^TM(pyridine) resin (17ml) post makes) and middle adding tri-n-butylamine (0.24ml, 1mmol).Vacuum concentrated solution then by adding continuously and evaporation anhydrous pyridine and dried residue, then adds again and evaporates two parts of 10mlN, dinethylformamide (DMF).

The synthetic of ribonucleoside triphosphote ester finished by following method.In single oligonucleotide (0.1mmol) solution of the anhydrous three fourth ammonium salts of the conduct in 1ml DMF or suspension, be incorporated in 1,1 among the 1mlDMF '-carbonyl dimidazoles (80mg, 0.5mmol).Blend compositions 30 minutes was inserted it in exsiccator under room temperature 4-12 hour then, then handled also with 33 μ l (0.8mmol) methanol and at room temperature reacted 30 minutes.Be incorporated in the pyrophosphoric acid three fourth ammoniums (0.5mmol) among the 5ml DMF, vigorous stirring was inserted mixture in the exsiccator under the room temperature about 24 hours then, made pyrophosphoric acid imidazoles precipitation.Sediment separate out; By centrifugal and in DMF resuspending, utilize 4 parts of 1ml DMF washing, obtain the purity of about 80-100%.Utilize isopyknic methanol to handle supernatant, vacuum evaporation is to doing.Residue contains chromatogram purification on the DEAE-cellulose column of linear gradient bicarbonate three fourth ammoniums (gradient is a 3L solution about 0 to 0.4M, pH5-7.5) at 2 * 20cm, collect fraction, identifies the fraction that contains the ribonucleoside triphosphote ester by spectrophotometry.The fraction that vacuum evaporation is suitable is dissolved in ribonucleoside triphosphote ester three second ammoniums in the methanol, to the concentration of about 0.05M, adds the sodium perchlorate acetone soln (15 equivalent) of 5 volumes, forms the sodium salt precipitation of ribonucleoside triphosphote ester.Should be appreciated that for those skilled in the art and utilize suitable precipitation, can make other salt of ribonucleoside triphosphote ester.By the salt of centrifugal collecting precipitation,, dry under vacuum with five phosphorous oxide with 4 parts of 1ml washing with acetones.

Other method of synthetic nucleosides triguaiacyl phosphate is available, comprises the method described in the following example.Embodiment 2: utilize 2,2,2-three bromomethyl phosphoro morpholino chloride synthetic nucleosides one-, two-and triguaiacyl phosphate

Basically utilize people's such as van Boom method (Trtrehedron Lett.32:2779-2782,1975), by one of single intermediate preparation ribonucleotide and derivant thereof-, two-and triguaiacyl phosphate.Usually; this reaction comprises simple function group reagent (2; 2; 2-three bromomethyl phosphoro morpholino chloride) with ribonucleotide (or derivatives thereof) reaction; generation have link to each other with ribonucleotide 2; 2, the phosphotriester derivant of 2-three bromomethyl blocking group (promptly prepare ribonucleotide 5 '-phosphoric acid morpholine ester (phosphomorpholidate) or ribonucleotide 5 '-phosphoric acid morpholine ester derivant).Utilize acid deblocking and neutralization, remove blocking group by the Cu/Zn coupling reaction, according to employed acid in the deblocking step and in neutralization procedure employed ammonium salt, generate one-, two-and triguaiacyl phosphate.That is to say,, can use HCl and ammonia in order to obtain the Monophosphate ribonucleotide; In order to obtain the bisphosphate ribonucleotide, can use one (three normal-butyl ammoniums) salt of phosphoric acid; In order to obtain the triguaiacyl phosphate ribonucleotide, can use two (three normal-butyl ammoniums) pyrophosphates.

General reaction is as shown in following:

Simple function group reagent (I) 2,2,2-three bromomethyl phosphoro morpholino chloride can be made by the following step: with 2,2,2-three bromomethyl phosphoro dichloride and morpholine react in absolute ether; Utilize method well-known in the art, separating obtained product is also used cyclohexane extraction/pentane recrystallization.Crystal 2,2, the muriatic fusing point of 2-three bromomethyl phosphoro morpholino is 79 ℃.

Under 20 ℃, in anhydrous pyridine, simple function group reagent (2mmol) was mixed 48 hours with 1mmol nucleoside or derivatives thereof; Then by chromatogram purification fraction reactant mixture (B.J.Hunt﹠amp; W.Rigby, Chem.﹠amp; Ind.1868,1967), obtain colourless nucleoside solid (III).In 20 ℃, in dry DMF, utilize the Cu/Zn galvanic couple to handle nucleotide 10 minutes, then filter to remove excessive Cu/Zn, obtain nucleoside phosphorylase morpholine ester (phosphoromorphlidate).

Utilize different acid and ammonia source to handle nucleoside phosphorylase morpholine ester then, obtain one-, two-or triguaiacyl phosphate.For Monophosphate, can utilize 0.01 N HCl, pH2 to handle down phosphoric acid morpholine esters 2 hours at 20 ℃, use ammonia (pH9) neutralization then and at Sephadex G-25 ^TMPurification on the post.

Similarly, nucleoside 5 '-triguaiacyl phosphate can obtain by following method: do not having under the condition of moisture, will react 3 hours down at 20 ℃ in the phosphoric acid morpholine ester (0.1mmol) in the 2ml dry DMF and two (the three positive fourth ammoniums) pyrophosphoric acids of the 0.5mmol in the 2ml dry DMF.Product is concentrated under vacuum, utilize Dowex 50 ^TM(ammonium form) handled; Utilize 3L linear gradient 0.0 to 0.3M Et ₂NH ₂CO ₃Solution, purification on 2 * 25cm DEAE cellulose column.

Nucleoside 5 '-bisphosphate can obtain by following method: do not having under the condition of moisture, in the 4ml anhydrous pyridine, phosphoric acid morpholine ester (0.1mmol) and 0.6mmol one (three positive fourth ammoniums) phosphoric acid were reacted 3 hours down at 20 ℃, and product carries out similarly concentrating and purification.In addition, in the pyridine solution that contains one (three positive fourth ammoniums) phosphoric acid, utilize the Zn powder to handle, phosphotriester derivant (III) directly can be transformed into corresponding nucleoside diphosphate ester.Just, do not having under the condition of moisture, 1mmol reagent III is being joined in the 15ml anhydrous pyridine solution that contains finely divided Zn powder of 0.1g and 12mmol one (three positive fourth ammoniums) phosphoric acid that is stirring, reacting 48 hours down at 20 ℃.Then the centrifugal Zn that makes of reactant mixture is become the ball shape, utilize per step 15ml water to steam supernatant altogether three times, then purification on the DEAE cellulose.Embodiment 3: synthetic 7-deazapurine 2 '-the dideoxyribonucleotide triphosphate ester

(1) synthetic 7-denitrogenation-2 '-deoxyinosine 5 '-the triguaiacyl phosphate triethyl ammonium salt.With 7-denitrogenation-2 '-deoxyinosine (25mg, 0.1mmol) be dissolved in trimethyl phosphate (250 μ l, 1.07mmol) in, add POCl ₃(18.5 μ l, 0.2mmol).Mixture was stirred 1.5 hours down at 0 ℃, be incorporated in two (three positive fourth ammoniums) the pyrophosphoric acid mixture of 0.5M in 1ml dry DMF and the 1ml three-n-butylamine then, vigorous stirring 1 minute; Utilize 1M Et ₂NH ₂CO ₃Aqueous solution is neutralized to pH7, and vacuum evaporation is to doing.Utilize linear gradient Et ₂NH ₂CO ₃, pH7 (11H ₂O/ll 0.7MTBK) solution, purification residue on 2.6 * 30cm DEAE-Sephadex post obtains colorless solid, UV (H ₂O) λ _MaxBe 258nm.

(2) synthetic 7-denitrogenation-2 '-deoxyadenosine 5 '-the triguaiacyl phosphate triethyl ammonium salt.By 7-denitrogenation-2 '-deoxyadenosine, can make this chemical compound similarly, obtain colorless solid, UV (H ₂O) λ _MaxBe 270nm.

(3) synthetic 7-denitrogenation-2 '-deoxyguanosine 5 '-the triguaiacyl phosphate triethyl ammonium salt.By 7-denitrogenation-2 '-deoxyguanosine, can make this chemical compound similarly, obtain colorless solid, UV (H ₂O) λ _MaxBe 259nm.Embodiment 4: Synthetic 2-chlorine deoxidation-3 adenosine phospho methylene biphosphonic acid esters (2CdAPMDP)

The nucleoside polyphosphate was synthesized, β in ribonucleoside triphosphote ester such as the deoxyadenosine triphosphate ester wherein, atom between γ-phosphorus atoms is substituted (people such as T.C.Myers by the atom that is not oxygen atom, Phosphonic analogues of nncleoside phosphate.I.The synthesis of 5 '-adenylyl methylene diphosphoaate, a phosphonic analogue of ATP.J.Am.Chem.Soc.85:3292-3295,1963; R.G.Yount, ATP analogues Adv.Enzymol.43:1-56,1975).This key stronger that has obtained being present in the nucleoside phosphorylase ester molecule that substitute than P-O key.

Synthetic is (J.Am.Chem.Soc.85:3292-3295,1963) of carrying out according to the method 1 or the method 2 of people such as Myers description basically.In method 1,2-chlorodeoxyadenosine 5 '-phosphoramidate and methylene biphosphonic acid reaction, the phosphonate analogue of generation nucleoside polyphosphate.In addition, utilize method 2, use dicyclohexyl carbodiimide (DCC) to be condensing agent, make 2-chlorodeoxyadenosine Monophosphate and methylene biphosphonic acid reaction.

According to method 1, methylenediphosphonate obtains by its tetra-ethyl ester of hydrolysis in dense HCl, and wherein tetra-ethyl ester is to react with excessive NSC 5284 by diiodomethane to make.Utilize 54ml newly to steam o-chlorphenol and handle 1,3-dicyclohexyl Guanidinium adenosine 5 '-phosphoramidate (3.6mmol) and methylenediphosphonate (10.8mmol); Use ice-cooled mixture, add the 36ml anhydrous pyridine.Under vibration frequently, gained solution was at room temperature placed 48 hours, at the ice-cold 300ml water that adds down.Utilize ether extraction solution 6 times (850ml altogether).Regulate aqueous solution to pH2 with 1N HCl, use 30g pickling Linesless charcoal (Norit A) to handle and stir then 30 minutes; Filter then and collect Linesless charcoal, water (5L altogether) washing up hill and dale.Nucleoside derivates by 35 ℃ of evaporations down, is concentrated into 400ml volume with eluent by concentrated ammonium hydroxide (3L altogether) eluting of 50% hydrate alcohol-5%.Spissated eluent is applied to 2.7cm * 31cm Dowex-2 ^TM(chloride; 8% is crosslinked) on the post; Use by mixing the linear gradient eluant solution that 2L 0.003N HCl (in blender) and 2L 0.003N HCl and 0.45N LiCl (in bin) make; Collect the fraction of 10ml; Use method well known in the art, utilize paper chromatography or uv absorption, identify the fraction that contains 2-chlorodeoxyadenosine-phospho methylene biphosphonic acid esters (2CdAPMDP).Utilize 1N LiOH neutralization to contain the fraction of 2CdATMDP and 30 ℃ of following evaporation and concentration; Handle spissated solution with 250ml acetone-10% methanol, be settled out solid; By this solid of centrifugalize, with the washing of acetone-10% carbinol mixture, till in cleaning mixture, can not detecting chloride.The 2CdATMDP lithium salts can be further purified: salt is dissolved in 100ml is adjusted in the water of pH8 with LiOH, utilize above-mentioned method, pass through Dowex-2 ^TMThis solution of column chromatography purification utilizes 1.5L0.003N HCl gradient solution eluting in mixing chamber, and utilizes the eluant solution of being made up of 1.5L 0.003N HCl and 0.45N LiCl in bin; Handle eluent as above-mentioned method, follow resolution of precipitate in 6ml water, it is precipitated with 40ml methanol.Be dissolved in the 15ml water final precipitate and lyophilizing, obtain 2CdATMDP four lithium salts powder.

Utilize method 2, methylenediphosphonate (11.4mmol) and 2-chlorodeoxyadenosine phosphonate ester (2.6mmol) are dissolved in pyridine (30ml) and the water (4ml), generate two-phase mixture; In adding DCC in 3 parts of aliquots (during the reaction beginning is 29mmol, is 48mmol after 4 hours, and is 19mmol after 12 hours) under the room temperature vigorous stirring.Afterreaction was finished in 24 hours, leached sedimentary 1,3-Dicyclohexylurea, washed with water.Water with the volume-adjustment of filtrate and cleaning mixture to cumulative volume 150ml, with ether extraction 5 times (300ml altogether).Solution is adjusted to pH8, at 2.5cm * 17.5cm Dowex-1 ^TMChromatogram purification on (formates, 2% is crosslinked) post; With this post of 1.5L water washing to remove pyridine.By constantly add this post of gradient solution eluting of the following solution acquisition that contains 500ml water in mixing chamber: 4N formic acid (500ml), 4N formic acid add that 0.1M ammonium formate (500ml) and 4N formic acid add 0.2M ammonium formate (1500ml), collect the 15ml fraction.By method well known in the art, utilize uv absorption to identify the fraction that contains 2CdATMDP (greatly in test tube 115-134).Fraction lyophilizing to the volume that contains 2CdATMDP that merges is approximately 200ml, uses 7g pickling Linesless charcoal (Norit A) to handle and stir then 15 minutes; Filter and collect Linesless charcoal, water (800ml altogether) washing.Product, removes by filter the trace Linesless charcoal and is lyophilized into powder by evaporating the volume that eluent is concentrated into 200ml down at 20 ℃ with 50% hydrate alcohol-5% concentrated ammonium hydroxide (600ml altogether) eluting.Powder dissolution in 4ml water, is utilized excessive 1M Barium acetate Treatment Solution; By centrifugal collection gained precipitation, wash with water and under 0 ℃, be dissolved among the 0.1N HBr.Utilize 1N NaOH regulator solution to pH6.5, by centrifugal collection gained precipitation, each sequentially washs with 2 * 2ml water, ethanol and ether.At room temperature use P ₂O ₄Drying sample 12 hours obtains two barium 2CdATMDP hydrates.Other nucleoside analog phospho methylene biphosphonic acid esters of the present invention can make similarly.Embodiment 5: utilize the EFAD mouse model, use the phosphate ester with anti-proliferative effect

Basically the method for describing according to embodiment 2 prepare one of anti-hypertrophy nucleoside analog-, two-and triguaiacyl phosphate.As above describing, these chemical compounds can form the local polyethylene cream, described cream use separately and contain 0.001gm% to 10gm% active component, PEG400 and Polyethylene Glycol 3350 in propylene glycol cream, thus formation unguentum mixture.Also can make to contain and strengthen the percutaneous permeability composition (as dimethyl sulfoxide or Azore ^TM) similar formulations.

In 6 hours to 5 day time,, the compositions of 10 μ l aliquots is applied to the dorsal part of EFAD hairless mouse with 6 hours intervals.When each interval finishes, kill the tissue that a mice and excision were treated, be placed on and contain ³In the tissue culture medium (TCM) of H-thymidine deoxyribose, carry out 2 hours pulse labelings, thereby measure one-, two-and triguaiacyl phosphate to the outgrowth effect of Skin Cell.Utilize conventional method from tissue, to isolate DNA then, measure by determining the radioactivity (cpm/ μ gDNA) in institute's separated DNA ³H-thymidine deoxyribose mixing in DNA.

Be used for suppressing one of outgrowth treatment psoriasis composition-, two-be to determine with the effectiveness of triguaiacyl phosphate by relatively mixing with radioactive mixing in the DNA of the tissue of crossing through the preparation for treating of different antipsoriatics to the DNA radioactivity of treated tissue not.

The total amount that is used for respectively testing the active component of Skin biopsy is that the administration number of times before measuring by the gm% of applied unguentum active component and mouse skin is determined.For the time point of each experiment, measure corresponding untreated time point by the veutro skin that uses same mouse. ³The difference of mixing of H-thymidine deoxyribose in dorsal part skin DNA and contrast veutro skin is used to calculate the total inhibition (percent) as the hyperplasia of the total flow function of phosphate ester of each time administration.

³The result that H-mixes also can be used for determining the inhibition percent (relatively dorsal part skin and veutro skin mixes) as the function of the time of each single test compositions.

As the function of time with as the function of administration active component total amount, to corresponding to among the DNA of the dorsal part skin of phosphate ester treatment ³H-mixes well below in the corresponding DNA that does not treat veutro skin ³H-mixes.Embodiment 6: use the polyamines biosynthesis in the zalcitabine triguaiacyl phosphate inhibition mouse model

It is to utilize phorbol ester 12-O-myristoyl phorbol-13-acetas (TPA by being determined at that the polyamines biosynthesis suppresses to measure; Be used for increasing the ODC activity) treat and before the amount that is present in the ODC Ornithine decarboxylase (ODC) of epidermis cell behind the mouse skin topical application zalcitabine triguaiacyl phosphate is carried out.

In brief, athymism baldness mice or EFAD mice can utilize TPA solution topical therapeutic, and described TPA solution can increase the ODC level in the epidermal tissue and simulate the level rising that is present in this kind of enzyme in the hypertrophy relevant with psoriasis usually.Treat in back 30 minutes to 1 hour at TPA, will contain as the zalcitabine triguaiacyl phosphate of active component and the unguentum that is used to strengthen the 0.01%DMSO of percutaneous permeability is applied to the skin area that TPA handled.Unguentum contains the zalcitabine triguaiacyl phosphate that makes according to the method among the embodiment 2 basically, and this unguentum is 5gm%, prepares in pharmaceutically acceptable cream in concentration, is applied to skin with the aliquot of 20 μ l; Other aliquot was per hour used once in 6 hours, used altogether 6 times.After 6 hours, utilize ODC level (people such as J.Lesiewicz, Modelsin Dermatology, vol.2 (H.I.Maibach ﹠amp in the technical measurement skin well known in the art; N.J.Lowe, eds.), 1985, pp112-116).In contrast, the not treatment skin of test athymism baldness mice is to determine normal ODC level; Test utilizes the skin of the athymism baldness mice that TPA treated, to determine to use the rising of ODC level behind the TPA.For further contrast, the skin that normal and TPA treated utilizes the unguentum that contains DMSO but do not contain the zalcitabine triguaiacyl phosphate to treat simultaneously.

To utilize TPA and contain ODC level in the skin of zalcitabine triguaiacyl phosphate unguentum treatment and the ODC level of normal skin and compare with ODC level in the skin that utilizes TPA to treat.The effectiveness of zalcitabine triguaiacyl phosphate treatment is by with respect to the reduction of the ODC level of TPA treatment skin and by with respect to the ODC level in the normal skin and definite.After the unguentum that utilization contains the zalcitabine triguaiacyl phosphate was treated, its ODC level had reduced significantly with respect to the ODC level of only not accepting the unguentum treatment through the TPA treatment or accepting not contain the unguentum treatment of zalcitabine triguaiacyl phosphate active component.

The Validity Analysis that it will be understood by those skilled in the art that similar treatment and utilize other di-deoxynucleoside phosphate ester to treat can be determined similarly.These di-deoxynucleoside phosphate esters comprise Didansine, dideoxy guanosine, DIDEOXYADENOSINE, didanosine and comprise nitrine Didansine (AZT) and the di-deoxynucleoside analog of dideoxy two dehydration thymidines one-, two-and triguaiacyl phosphate.Embodiment 7: utilize and transplant the mice that the psoriasis application on human skin is arranged, application has the floxuridine of treatment proliferative effect and the phosphate ester of 5-floxuridine

When the part is applied to be transplanted to people's psoriasis skin on the athymic mouse, Skin biopsy is used to detect the anti-proliferative effect of floxuridine triguaiacyl phosphate and 5-fluorouridine triphosphate ester.Utilize technology well known in the art, diameter is approximately the dorsal part of people's psoriasis skin transplantation of 1-10mm to athymic mouse.After graft fully heals, utilize contain active component for floxuridine triguaiacyl phosphate, 5-fluorouridine triphosphate ester or one-or the antipsoriatic treatment of bisphosphate suffer from psoriasic application on human skin.These chemical compounds are to make according to the method that embodiment 1 describes on substantially.Pharmaceutical composition is by comprising that being present in as the disclosed local concentration with active component in the polyethylene cream and active component of early stage document is that 0.1gm% to 10gm% makes.In 1-15 days, with 6 hours intervals, will contain the parts of skin that the unguentum of active component is applied to transplant with 20 μ l aliquots.Behind 24 hours treatment stages, the skin of checking treatment is to determine psoriasic rough labelling (redness of the agile and inflammation of skin).After 15 days, skin is separated from mice; Utilize conventional histological techniques,, determine the quantity and the expansible relative extent of the capillary tube in the skin of cuticular cellulose by biopsy by microscopic observation.

With the quantity of cuticular cellulose be transplanted to similar detection on the athymism baldness mice normally and do not treat people's psoriasis skin and compare, the effect of determining to utilize the pharmaceutical composition that contains floxuridine triguaiacyl phosphate and 5-fluorouridine triphosphate ester to treat.By the quantity of neutrophilic leukocyte and monocyte in definite epidermal hair tubule under suitable dyeing condition, the inflammation relative extent is quantitative in can organizing.Can discover with respect to the capillary tube relative expansion degree of not treating psoriasis skin and normal control skin.

The quantity of cellular layer, degree of inflammation and capillary tube degrees of expansion are that the amount of the active component determined with the natural law of treatment and by composition concentration and dosage is corresponding in the epidermis.With respect to untreated psoriasis skin, contain floxuridine triguaiacyl phosphate, 5-fluorouridine triphosphate ester or one-or the compositions of bisphosphate all demonstrate the remarkable reduction of psoriasis symptom.Embodiment 8: suffer from the psoriasic application on human skin use 7-deazapurine 2 with treatment proliferative effect '-the dideoxyribonucleotide triphosphate ester

Utilization comprise basically as synthetic 7-denitrogenation-2 as described in the embodiment 3 '-deoxyinosine 5 '-triethyl ammonium salt of triguaiacyl phosphate is as the composite cream of active component, peel off the volunteer of skin speckle and treat on two forearms, having psoriasis.The concentration of the active component in pharmaceutically acceptable unguentum such as early stage document in the disclosed Polyethylene Glycol unguentum is 50gm%.

Peeling off and inflammation on macroscopy two skin of arm, utilization contain 7-denitrogenation-2 '-deoxyinosine 5 '-cream of triguaiacyl phosphate writes down result's (written visual report and photo) before treating.In 14 days time, with 6 hours intervals, about diameter that one or two unguentum is applied on one of them forearm is the psoriasis skin area of 3cm.Another forearm during 14 days in maintenance do not treat.

In 14 days, with the naked eye check peeling off and inflammation of skin on two arms every day, the record result carries out the both sides paired comparison.If volunteer has the psoriasis speckle of q.s, can test simultaneously contain the 7-denitrogenation-2 that concentration is 50gm% '-deoxyadenosine 5 '-triguaiacyl phosphate and 7-denitrogenation-2 '-deoxyguanosine 5 '-analogous composition of the triethyl ammonium salt of triguaiacyl phosphate treats psoriasic activity.

Last at 14 days, as the method for describing among the embodiment 7, the corium and the epidermis that use conventional organization method to take out the 1-2mm diameter partly carry out biopsy and micrography (determining the quantity of cellular layer in the epidermis, the inflammation degree in the corium and capillary tube degrees of expansion).Never treat and take out skin in the psoriasis skin of arm as negative contrast, never treat in the arm and to take out normal skin and contrast as the front, from utilization contain 7-deazapurine-2 '-effectiveness of the definite active component with respect to control sample of skin conduct is taken out in zone that the unguentum of dideoxyribonucleotide triphosphate ester treat.

The naked-eye observation in zone shows in pairs in the treatment psoriasis skin that carries out on the identical volunteer and the both sides of not treating psoriasis skin: in 14 days, utilize 7-deazapurine-2 '-psoriasis symptom in the skin of dideoxyribonucleotide triphosphate ester treatment significantly reduces.The result of naked-eye observation has obtained by treatment and the not micrographic support of the skin of area for treatment taking-up.Similarly, one-and bisphosphate also demonstrated activity.Embodiment 9: use the 2-chlorodeoxyadenosine phosphate ester with treatment proliferative effect on the skin that peels off the skin mice

Utilization contain be basically as embodiment 2 described methods make as one of the 2-chlorodeoxyadenosine of active component-, two-or the medicine composite for curing of triguaiacyl phosphate demonstrate and have a psoriasic age of struvite epidermis and be 35-42 days inbreeding fsn/fsn mice.Said composition contain concentration be one of 5gm%-, two-or triguaiacyl phosphate active component.Compositions is applied to the 1-5mm part of the veutro skin of fsn/fsn mice with 5 μ l aliquots, with 12 hours intervals; Contrast fsn/fsn mice is also similarly with not containing the composition of active components treatment.Begin after 12 hours to observe skin roughly and microscopically every day in the administration first time, after extending to 15 days.Utilize conventional Dermatology and Histological method, observed result is used for epidermal hyperplasia degree and the inflammation feature of determining that psoriasis skin occurs.

For the treatment after each observation station, the treatment skin with do not treat same mouse similar portions and with only with carrier components without one of 2-chlorodeoxyadenosine-, two-or the skin that triguaiacyl phosphate is treated compare.Can observe after a few days: compare with untreated skin or through not containing the skin that composition of active components treats, utilize contain one of 2-chlorodeoxyadenosine-, two-or the inflammation of the compositions of the triguaiacyl phosphate parts of skin for the treatment of and peel off remarkable improvement.

By utilization contain 2-chloro-2 '-fluorine arabinose deoxyadenosine one-, two-or the fsn/fsn mice treated as composition of active components of triguaiacyl phosphate, can similarly analyze.In 14 days, use these compositionss, can observe similar psoriasis symptom and reduce.Embodiment 10: use the 6-aza uridine triguaiacyl phosphate with treatment proliferative effect on the psoriasis application on human skin

Utilization comprises basically as the unguentum of synthetic 6-aza uridine triguaiacyl phosphate as described in the embodiment 1 as active component, the volunteer that has psoriasis skin speckle on arm and lower limb is treated.The concentration of the active component in pharmaceutically acceptable unguentum such as polyethylene glycol groups unguentum is 25gm%.

Peeling off and inflammation on macroscopy arm and the lower limb skin write down observed result (written visual report and photo) before the unguentum that utilization contains 6-aza uridine triguaiacyl phosphate is treated.In 14 days time, with 3 hours at interval, be applied to the quantitative aliquot that respectively is 50 μ l on one of them arm and psoriasis skin area that wherein diameter on the one leg is about 5-10cm.As negative contrast, another arm and lower limb during 14 days in maintenance do not treat.

In 14 days, with the naked eye check peeling off and inflammation of skin on two arms and two lower limbs every day, the record result carries out the both sides paired comparison.

Last at 14 days, as the method for describing among the embodiment 7, skin (corium and the epidermis) part of utilizing conventional organization method to take out the 1-2mm diameter is carried out biopsy and micrography, determines cellular layer quantity, the inflammation degree in the corium and capillary tube degrees of expansion in the epidermis.Never take out skin in the psoriasis skin of area for treatment and/or lower limb as negative contrast, never take out normal skin in the area for treatment and contrast, contain zone taking-up skin that the unguentum of 6-aza uridine triguaiacyl phosphate treat effectiveness with definite active component with respect to control sample from utilization as the front.

The naked-eye observation in zone shows in pairs in the treatment psoriasis skin that carries out on the identical volunteer and the both sides of not treating psoriasis skin: in 14 days, utilize the psoriasis symptom in the skin of 6-aza uridine triguaiacyl phosphate treatment significantly to reduce.The result of naked-eye observation has obtained by treatment and the not micrographic support of the skin of area for treatment taking-up.One of 6-aza uridine-also can obtain similar result with bisphosphate.Embodiment 11: utilize 2-chlorodeoxyadenosine phospho methylene biphosphonic acid esters (2CdATMDP) treatment people psoriasis skin

The outgrowth effect of treatment of 2CdATMDP when being transplanted to people's psoriasis skin on the athymism baldness mice and being used to detect topical application.Naked-eye observation and the analysis of microscope Skin biopsy are used to detect the effectiveness for the treatment of psoriasis composition.

Utilize technology well known in the art, diameter is approximately people's psoriasis skin transplantation of 1-10mm to the dorsal part of athymic mouse.After graft fully heals, utilize the treatment psoriasis medicine treatment that contains active component nucleoside analog 2CdATMDP (method of being described by embodiment 4 makes basically) to suffer from psoriasic application on human skin.Pharmaceutical composition comprises that being present in is about 0.5gm% to 5gm% as the disclosed local concentration with active component in the polyethylene cream and active component of early stage document basically.In 1-30 days, with 12 hours intervals, will contain the parts of skin that the unguentum of active component is applied to transplant with 10 μ l aliquots.In 24 hours treatment stages, the skin of checking treatment is to determine psoriasic rough indication (as peeling off and inflammation).When 15 days and 30 days, skin is separated from the transplanting zone of mice; Utilize conventional histologic analysis to determine the quantity of cuticular cellulose and the expansible relative extent of capillary tube in the dermis of skin.

With the quantity of cuticular cellulose be transplanted to similar detection on the athymism baldness mice normally and do not treat people's psoriasis skin and compare, the effect of determining to utilize the pharmaceutical composition that contains 2CdATMDP to treat.Except the expansible relative extent of capillary tube, by determining the quantity with respect to neutrophilic leukocyte and monocyte in the dermal capillaries of not treating psoriasis skin and normal control skin, inflammation is quantitative in can organizing.

The amount of the natural law of the quantity of cellular layer, degree of inflammation and capillary tube degrees of expansion and treatment and (being determined by composition concentration and dosage) active component is relevant in the epidermis.After whole treatment stage, compare the remarkable reduction that in the transplanting psoriasis skin of treatment, can observe inflammation and peel off with the not treatment contrast psoriasis skin in being transplanted to athymism baldness mice.

Utilization comprises the pharmaceutical composition of nucleoside analog 2CdATMDP, a forearm of the volunteer that has psoriasis skin speckle on two forearms is treated similarly twice of every day (50 μ l are used in each treatment).The state of an illness of the treatment psoriasis skin that every day observation and another forearm control zone (do not treat skin or utilization and do not contain the skin that the ointment preparation of active component is treated) are compared, took out the skin living tissue of diameter at the 15th day and the 30th day, analyse according to the above-described normal structure credit that is used for mouse model and detect for about 1-3mm.After 15 days, and do not treat skin or utilize the inert composition of preparation just and skin that the non-activity composition is treated is compared, the inflammation that observes on the skin that utilizes the compositions that contains 2CdATMDP to treat alleviates.After 30 days, compare, utilize the inflammation that observes in the skin that contains the composition of active components treatment and peel off significantly to change with the skin contrast speckle of another forearm.Embodiment 12: utilize 2-chloro-2 '-fluorine arabinose deoxyadenosine phosphate ester treatment people's local dermatitis

Basically as embodiment 2 described method Synthetic 2-chloro-2 '-fluorine arabinose deoxyadenosine one-, two-or triguaiacyl phosphate.Utilization contain each concentration for the 2-chloro-2 of 5gm% '-fluorine arabinose deoxyadenosine one-, two-and the washing liquid combination treatment of the mixture of triguaiacyl phosphate suffer from the adolescence patient of local dermatitis; This part dermatitis unknown cause is a feature with extreme pruritus and eczema damage on fist and neck.This washing liquid is applied to the infected zone of skin, can use about at every turn 1-3cc every day nearly 12 times.After 1-4 week treatment, when beginning to occur, significantly reduce with the skin pruritus and the inflammation of dermatitis with respect to the patient.Further, after treatment, the eczema damage on the treatment skin just will be healed or heal.Thus, this washing liquid only just need be used when pruritus recurs.Embodiment 13: utilize di-deoxynucleoside phosphate ester treatment lichen planus

Basically as one of the synthetic zalcitabine of embodiment 2 described methods-, two-or triguaiacyl phosphate.The phosphate ester of each zalcitabine is mixed with the composite ointment based on Polyethylene Glycol that concentration is 0.5gm%.To suffer to have pimple inflammation and pruritus erythra on its arm and the lower limb is that the volunteer of the chronic lichen planus of feature is divided into three groups, uses a kind of combination treatment for every group: one of zalcitabine-, two-or triguaiacyl phosphate.Every group is used unguentum only to use on an arm or lower limb, stays another arm or lower limb and does not treat, to carry out and the both sides contrast for the treatment of skin.7 days to 3 months during, used at interval the unguentum of about 1cc volume in every about 6 hours.With the naked eye check treatment and untreated regional two weeks every day, check once or check as required (doctor by treatment determines) afterwards at least weekly.As if when inflammation and/or pimple form when reducing, the number of applications that the treatment doctor also can reduce every day as required is to keep control disease.

When as if as if the inflammation of area for treatment reduce and pruritus erythra when reducing, take out skin living tissue (approximately 1-3mm) from the treatment limbs of the infected zone of volunteer with not treating limbs and carry out the histological examination of (as the quantity of epidermal area and the albescent line relevant or the sign of point) of inflammation (as the dermal capillaries expansion) and pimple feature with pimple.

For each group in these three groups, compare with corresponding skin of not treating limbs, demonstrated significant improvement in the inflammation of the lichen planus of some members of this group on the skin of its treatment arm or lower limb and the pruritus erythra.

It will be understood by those skilled in the art that the similar thing of other nucleotide phosphate of the present invention and comprise that the similar phosphate ester of the dideoxy chemical compound of Didansine, dideoxy guanosine, DIDEOXYADENOSINE, didanosine and di-deoxynucleoside analog (comprising ATZ and d4T) can be used for pharmaceutical composition similarly.Embodiment 14: utilize acyclovir phosphate ester treatment actinic keratosis

Basically as synthetic acyclovir triguaiacyl phosphate as described in the embodiment 1.Utilize method well known in the art to prepare composite ointment, said composition contains 0.1-0.5gm% acyclovir triguaiacyl phosphate and can strengthen about 0.01gm%Azone of percutaneous permeability ^TM

One group of middle age volunteer that has the actinic keratosis speckle on the skin (as on nose, hands, arm or the lower limb) that is exposed under the sun is divided into three groups: one group is utilized the liquid nitrogen freezing actinic keratosis and cell killing; One group is utilized acyclovir triguaiacyl phosphate compositions to treat; One group is matched group, utilizes to contain inert fraction and Azone ^TMAnd the compositions that does not contain the acyclovir triguaiacyl phosphate is treated.

In time of one month nearly, utilize the group of the composite ointment treatment that contains or do not contain the acyclovir triguaiacyl phosphate to apply unguentum twice (morning and evening) every day, the administration volume is enough to cover infected zone (approximately 0.01-0.5cc).Every other day with the naked eye check the skin and the hourly observation result (written and photo) of every group of treatment by the clinician.After the treatment, volunteer was examined the recurrence situation of area for treatment actinic keratosis every 6 months.

For the group of unguentum treatment that utilization contains the acyclovir triguaiacyl phosphate, a large amount of patients demonstrate the reduction of keratinization growth, and the redness in a month postoperative infection zone of treatment weakens.The clinician can determine whether that needs continue platform and treat according to 6 months inspection.Those skilled in the art are to be understood that, one of acyclovir-and bisphosphate also can be used in the similar pharmaceutical preparation, and ganciclovir or other anti-hypertrophy nucleoside analog of the present invention one-, two-also can be used for treating in excessively relevant with the hypertrophy dermopathic compositions with anti-proliferative activity with triguaiacyl phosphate.

For the group of utilizing the unguentum treatment that does not contain the acyclovir triguaiacyl phosphate, the observation actinic keratosis does not have any variation after 1 month, then the infected zone is utilized clinical acceptable method to remove.

For the group of utilizing the liquid nitrogen freezing actinic keratosis, the cell of all area for treatment is all dead in 24 hours, has then formed the slough of dead cell, and follow pain in agglutination.In some cases, see cicatrix in area for treatment.

Claims (40)

1. Use of 2-chlorodeoxyadenosine monophosphate or a pharmaceutically acceptable salt thereof in the preparation of a medicament for the treatment of mammalian diseases characterized by excessive proliferation of skin cells, said treatment by treating mammals This is done by topical application of an effective amount of the drug to the diseased skin area. 2. Use of 2-chlorodeoxyadenosine monophosphate or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for treating a mammal suffering from psoriasis by topically applying an effective amount of the medicament to the diseased skin area of the mammal conduct. 3. 2. The use of claim 1 or 2, wherein said medicament comprises 2-chlorodeoxyadenosine monophosphate or a pharmaceutically acceptable salt thereof in a carrier suitable for topical administration at a concentration of about 0.001 gm% to about 100 gm%. 4. Use of 2-chloro-2'-arabino-fluoro-2'-deoxyadenosine monophosphate or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating a diseased mammal characterized by Skin cell hyperproliferation by topical application of an effective amount of said medicament to an area of diseased skin in a mammal. 5. The use of 2-chloro-2'-arabino-fluoro-2'-deoxyadenosine monophosphate or its pharmaceutically acceptable salt in the preparation of medicines for the treatment of mammals suffering from psoriasis. Topical application of an effective amount of the drug to a diseased skin area is performed. 6. The use of claim 4 or 5, wherein the medicament comprises 2-chloro-2'-arabino-fluoro-2'-deoxyadenosine at a concentration of about 0.001 gm% to about 100 gm% in a carrier suitable for topical administration Glycoside monophosphate or a pharmaceutically acceptable salt thereof. 7. Use of 2-chlorodeoxyadenosine phosphate or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for the treatment of a diseased mammal characterized by hyperproliferation of skin cells by means of This is done by topical application of an effective amount of the drug to the diseased skin area. 8. Use of 2-chlorodeoxyadenosine phosphate or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for treating a mammal suffering from psoriasis by topical application of an effective amount of said medicament to a diseased skin area of the mammal . 9. The pharmaceutical application of 2-chlorodeoxyadenosine diphosphate or its pharmaceutically acceptable salt according to claim 7 or 8. 10. The pharmaceutical application of 2-chlorodeoxyadenosine triphosphate or its pharmaceutically acceptable salt according to claim 7 or 8. 11. Use of 2-chloro-2'-arabino-fluoro-2'-deoxyadenosine phosphate or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for the treatment of a diseased mammal characterized by skin Cellular hyperproliferation, said treatment being carried out by topical application to the diseased skin area of a composition containing said drug. 12. Use of 2-chloro-2'-arabino-fluoro-2'-deoxyadenosine phosphate or a pharmaceutically acceptable salt thereof in the preparation of a medicament for the treatment of mammals suffering from psoriasis, said treatment by treating mammals This is done by topical application of an effective amount of the drug to the diseased skin area. 13. The pharmaceutical application of 2-chloro-2'-arabino-fluoro-2'-deoxyadenosine diphosphate or its pharmaceutically acceptable salt according to claim 11 or 12. 14. The pharmaceutical application of 2-chloro-2'-arabino-fluoro-2'-deoxyadenosine triphosphate or its pharmaceutically acceptable salt according to claim 11 or 12. 15. 2-chlorodeoxyadenosine phosphate, 2-chloro-2'-arabino-fluoro-2'-deoxyadenosine phosphate according to claim 1, 4, 7 or 10, pharmaceutically acceptable salts of said phosphates, Or the use of a mixture of said phosphate ester and said salt, wherein the hyperproliferative skin disease to be treated is selected from the group consisting of atopic dermatitis, lichen planus, actinic keratoses, basal cell carcinoma and squamous cell carcinoma. 16. 2-chloro-deoxyadenosine phosphate, 2-chloro-2'-arabino-fluoro-2'-deoxyadenosine phosphate of claim 1, 2, 4, 5, 7, 8, 11 or 12, said Pharmaceutical use of a pharmaceutically acceptable salt of a phosphate ester, or a mixture of said phosphate ester and said salt, wherein the affected individual is a human. 17. A pharmaceutical formulation comprising an effective antiproliferative amount of 2-chlorodeoxyadenosine monophosphate, or a pharmaceutically acceptable salt thereof, in a carrier suitable for topical administration. 18. A pharmaceutical formulation comprising an antipsoriatic effective amount of 2-chlorodeoxyadenosine monophosphate, or a pharmaceutically acceptable salt thereof, in a carrier suitable for topical administration. 19. The pharmaceutical formulation of claim 17 or 18, wherein the concentration of 2-chlorodeoxyadenosine monophosphate or a pharmaceutically acceptable salt thereof is about 0.001 gm% to 50 gm%. 20. The pharmaceutical formulation of claim 17 or 18, wherein the concentration of 2-chlorodeoxyadenosine monophosphate or a pharmaceutically acceptable salt thereof is about 0.01 gm% to 10.0 gm%. twenty one. The pharmaceutical formulation of claim 17 or 18, wherein the concentration of 2-chlorodeoxyadenosine monophosphate or a pharmaceutically acceptable salt thereof is about 0.1 gm% to 1.0 gm%. twenty two. The pharmaceutical formulation of claim 17 or 18, wherein the carrier comprises an aqueous cream. twenty three. The pharmaceutical preparation according to claim 17 or 18, further comprising propylene glycol, polyethylene glycol 400 or polyethylene glycol 3350. twenty four. The pharmaceutical formulation according to claim 17 or 18, further comprising an ingredient which enhances skin penetration when applied topically. 25． A pharmaceutical formulation comprising an effective anti-proliferative amount of 2-chloro-2'-arabino-fluoro-2'-deoxyadenosine monophosphate or a pharmaceutically acceptable salt thereof in a carrier suitable for topical administration. 26． A pharmaceutical formulation comprising an antipsoriatic effective amount of 2-chloro-2'-arabino-fluoro-2'-deoxyadenosine monophosphate or a pharmaceutically acceptable salt thereof in a carrier suitable for topical administration. 27． The pharmaceutical formulation of claim 25 or 26, wherein the concentration of 2-chloro-2'-arabino-fluoro-2'-deoxyadenosine monophosphate or a pharmaceutically acceptable salt thereof is about 0.001 gm% to 50 gm%. 28． The pharmaceutical formulation of claim 25 or 26, wherein the concentration of 2-chloro-2'-arabino-fluoro-2'-deoxyadenosine monophosphate or a pharmaceutically acceptable salt thereof is about 0.01 gm% to 10.0 gm%. 29． The pharmaceutical formulation of claim 25 or 26, wherein the concentration of 2-chloro-2'-arabino-fluoro-2'-deoxyadenosine monophosphate or a pharmaceutically acceptable salt thereof is about 0.1 gm% to 1.0 gm%. 30． The pharmaceutical formulation of claim 25 or 26, wherein the carrier comprises an aqueous cream. 31． The pharmaceutical preparation according to claim 25 or 26, further comprising propylene glycol, polyethylene glycol 400 or polyethylene glycol 3350. 32． The pharmaceutical formulation of claim 25 or 26, further comprising an ingredient that enhances skin penetration when applied topically. 33． A pharmaceutical formulation comprising an effective antiproliferative amount of 2-chlorodeoxyadenosine phosphate, or a pharmaceutically acceptable salt thereof, in a carrier suitable for topical administration. 34． 33. The pharmaceutical formulation of claim 33 comprising an effective anti-proliferative amount of 2-chlorodeoxyadenosine diphosphate or a pharmaceutically acceptable salt thereof in a carrier suitable for topical administration. 35． 33. The pharmaceutical formulation of claim 33 comprising an effective antiproliferative amount of 2-chlorodeoxyadenosine triphosphate or a pharmaceutically acceptable salt thereof in a carrier suitable for topical administration. 36． A pharmaceutical formulation comprising an effective anti-proliferative amount of 2-chloro-2'-arabino-fluoro-2'-deoxyadenosine phosphate or a pharmaceutically acceptable salt thereof in a carrier suitable for topical administration. 37． The pharmaceutical formulation of claim 36 comprising an effective anti-proliferative amount of 2-chloro-2'-arabino-fluoro-2'-deoxyadenosine diphosphate or a pharmaceutically acceptable salt thereof in a carrier suitable for topical administration . 38． The pharmaceutical formulation of claim 36 comprising an effective anti-proliferative amount of 2-chloro-2'-arabino-fluoro-2'-deoxyadenosine triphosphate or a pharmaceutically acceptable salt thereof in a carrier suitable for topical administration . 39． The pharmaceutical formulation of claim 33, wherein the concentration of 2-chloro-deoxyadenosine phosphate or a pharmaceutically acceptable salt thereof is about 0.001 gm% to 100 gm%. 40． The pharmaceutical formulation of claim 36, wherein the composition concentration of 2-chloro-2'-arabino-fluoro-2'-deoxyadenosine phosphate or a pharmaceutically acceptable salt thereof is about 0.001 gm% to 100 gm%.