JPH05506842A - How to treat neurodegenerative conditions - Google Patents

Abstract

(57) [Summary] This bulletin contains application data before electronic filing, so abstract data is not recorded.

Description

[Detailed description of the invention] How to treat neurodegenerative conditions Application cross-reference This application is filed under CI No. 407.913, filed September 15, 1989. It is P.

Background of the invention The present invention provides a method for treating neurodegenerative conditions based on increasing extracellular concentrations of adenosine. The purpose is to explain how to place the item.

The etiology of major neurodegenerative diseases remains unknown. Such diseases, such as Burkis Huntington's disease, Amyotrophic lateral sclerosis (ALS) (Lou Gehrig's Diseases), and Alzheimer's disease treatment is recognized as difficult: slowing or inhibiting this degenerative process. Few, if any, treatments have been found to be effective.

Parkinson's disease is a major neurodegenerative disease that commonly affects the elderly. above As such, the specific pathogenesis is not fully understood.

However, Parkinson-like syndromes are induced by exposure to certain chemical messengers. There are cases. Such substances include methamphetamine and l-methyl-4-phthalate. Two substances, phenyl-1,2,3,6,-tetrahydropyridine (MPTP), , has been used as a model to study Parkinson's disease.

Parkinson's disease is characterized by disorders within the brain, specifically those affecting the striatum; and dopamine depletion, especially in the striatum (nuclei of the basal ganglia, especially the substantia nigra, putamen and caudate nucleus) It is characterized by depletion in . Dopamine in individuals affected by Parkinson's disease In an attempt to alleviate depletion, it has been used as a therapeutic agent to alleviate the symptoms of Parkinson's disease. It is a precursor of dopamine that can pass through the blood-brain barrier relatively well. started to be used. The goal is to improve the overall effect of L-dopa. is a peripheral decarboxylase inhibitor that reduces L-dopa metabolism in peripheral tissues. Carbidopa is often used.

When either methamphetamine or MPTP is administered systemically to experimental animals, the punctate striae degenerative changes that occur in the body's dopaminergic neurons or their axon terminals was found. Both methamphetamine and MPTP are associated with striatal dopamine (D A) and also decrease tyrosine hydrokindase (TH) activity, and Histochemical findings of nerve terminal degeneration in the neostriatum are observed. these nerves Some of the effects of degeneration are associated with overactivity of excitatory amino acid (EAA) neurotransmission. It was hypothesized that this may be the case. N-methyl, one of the EAA receptors Treatment with a non-competitive blocker of D-aspartate (NMDA) receptors , DA amount and TH activity induced by methamphetamine or MPTP administration. It has been shown to partially antagonize the NMDA-mediated decrease in sex. Based on these findings It is hypothesized that EAA is related to neurodegenerative symptoms such as Parkinson's disease. Ta. (Sonsa, 11a, P, K, et al. 5science 243:398 -400 (1989), methamphetamine-induced nigrostriatal dopamine production (See ``The Role of Excitatory Amino Acids in Neurotoxicity'').

Animals were treated with methamphetamine or MPTP to simulate the effects of Parkinson's disease. treatment and created a model of Parkinson's disease. These experimental models are It has been used to evaluate the efficacy of various treatments for cancer disease.

Administration of MTPTP to animals provides a useful Parkinson's model. M The end result of PTP administration is the neocorticolimbic region, a subcortical region in the center of the brain. destruction of the striatum in the brain, an area that is damaged in Parkinson's disease. . The neurotransmitter dopamine is concentrated within the striatum. Parkinson's disease is It is characterized by damage to this area of the brain and depletion of dopamine levels. some kind In (primates), degeneration of the striatum is accompanied by behavioral symptoms similar to human Parkinson's disease. It was reported that

Methamphetamine also compromises the striatum, but is slightly less selective than MPTP. , seems to induce striatal degeneration by a mechanism different from that of MPTP.

adenosine, 9-β-D-ribofuranosyl adenine (nucleo of purine adenine) ) belong to a biochemical class called purine nucleosides, and include Fox and Ke Annual Reviews of Biochemistry, An important biochemical cell regulatory molecule described in Volume 47, Page 635, 197g. be.

Adenosine interacts with a wide range of cell types and is involved in a myriad of biological effects. There is. Adenoone is an inhibitory neuromodulator odulator) in the brain [5nyder , S.H., ,Ann, Rev. Neural Sci., 8:103-12419. 85, Neurosci and Biobehav of Marangos et al. Rev, 9:421-430 (1985), Dunviddie, Int, R. ev.

Neurobiol, 27: 63-L30 (1985)]. This effect is mediated by extracellular receptors [Regula of Londos et al. tory Functions of Aden.

5jne, pp. 17-32 CBerne et al. (1983). against nervous tissue Among the established effects of adenosine on neuroinflammation are inhibition of neuroinflammation [pHllis et al. Europ, J. Pharmacol, 30:125-129 (1 975)] and inhibition of calcium-dependent neurotransmitter release [Dunwidd ie, 1985]. Behaviorally, adenosine and its metabolically stable amino acid Analogs are effective through specific adenosine receptor antagonists. deep anticonvulsant action that is radically reversed vulsant) and has sedative effects [Dunwiddie et al. J, P harmacol, and Exptl, Therapeut, 22 0 Near 0-78 (1982); Radulovacki et al. J, Pharm acol, Exptl, Thera, 228:268-274 (19 8])]B In fact, adenosine serves as a natural anticonvulsant and its extracellular levels Modifying substances have been proposed as modulators of seizure activity. [Dragunow et al. Epilepsia 2B+480-487 (1985 ); Lee et al. Brain Res, 21: 1650-164 ( 1984)]. Additionally, adenosine is a powerful vasodilator and inhibitor of immune cell function. bitters, inhibitors of granulocyte oxygen free radical production, anti-arrhythmic agents, and It is a toxic neuromodulator. Considering its broad spectrum of biological activity, adenosin Significant efforts have been made to establish ways to use the drug and its analogues in actual treatment. being paid.

Adenosine binds to receptors anchored to the plasma membrane, thereby binding to its membrane receptors. Previous research has shown that azoncin acts on the bloodstream. Attempts have also been made to increase the extracellular concentration of adenosine by administering it. However, a Effective extracellular denosine must be administered to the patient to maintain therapeutic concentrations. Adenosine adenosine is toxic, so administration of adenosine alone is not effective for its treatment. There are limited routes. Additionally, adenosine receptors expose adenosine This can lead to negative feedback mechanisms such as receptor downregulation. receive your blessings.

Other methods exist to achieve high local extracellular concentration effects of adenosine, and research is ongoing. It has also been investigated. (a) reagents that specifically block adenosine transport; [Annals of Paterson et al. of the New York AcadeIIly or 5science s, vol. 255, p. 402 (1975); and Life by Deckert et al. 5science, Volume 42, Pages 1331-1345], (b) Decomposition of adenosine Prevention (-Carton and Seega+1ller's The Journal f　C11nical Investigation, Volume 57, Page 274 (19 76) co, and (c) azo/syn azo which selectively binds to adenosine receptors. There is the use of analogues.

Compounds that selectively increase extracellular adenosine stimulate cells in the hippocampus associated with memory. It may also be used as a plaster for preventive protection. The hippocampus has more neurons than any other brain region. It has denosine and glutamate receptors. Therefore, I will explain below This is the most susceptible to cerebral hypoplasia or stroke. It is. Several recent studies have shown that Alzheimer's disease is a chronic, asymptomatic brain deficiency. It has been shown that blood may be the cause. Therefore, it selectively reduces extracellular adenosine levels. Compounds that increase brain damage may be used to treat and/or treat both clear stroke and Alzheimer's disease. or can be used to prevent.

What is currently recognized is that brain damage occurs over a relatively short period of time (on the order of 2-8 minutes). The blood triggers a series of events that result in the death of a selected population of neonates in the brain. Rub. This process is delayed excitotoxic. excitatory amino acids such as glutamate and aspartate It is caused by an ischemia-induced increase in transmitter release. After stroke, Within hours to days, certain neurons in the brain undergo metabolic exhaustion and death. is overstimulated by EAA. Excess released glutamate can cause stroke microscopic Glutamate receptors in the brain are thought to be the main factor associated with cell damage. Inhibition would be beneficial in stroke treatment. In animals, glutamate receptors blockers have been shown to be effective in alleviating and preventing stroke-related neurological damage. is shown. However, inhibition of these receptors lacks specificity and leaves much to be desired. It has been shown to cause unwanted side effects. Excita of Church et al. tory Am1no Ac1d Transmission'', H, 5-1 Page 18 (Alan R, Li5s, Inc, 1987).

Adenosine has been shown to be a potent inhibitor of glutamate release in the brain. Ru. The CA-I region of the brain is selectively susceptible to post-stroke destruction. 1 and 3 days after stroke The study observed on day 6 and 6 showed that the CA, -I region of the hippocampus was gradually destroyed over time. However, cyclohexyladenosine (CHA) When azo/syn agonists are given for a short period of time after stroke, the CA-I region is Significantly protected [Daval et al., Brain Res, 491:212 -226 (1989) and Mar'angos.

Med, Hypothesig 32:45-49(1,990)]. animal raw Beneficial results were also observed in survival rates. However, due to its global impact, CHA and have non-specific side effects. For example, it can undesirably lower blood pressure and Decreases heart rate and significantly increases blood glucose.

Hyperglycemia has been reported to be associated with poor prognosis in stroke [Helgason , 5troke 19(8):1049-1053(1!J88)]. moreover, Moderate hypoglycemia induced by insulin treatment is associated with experimentally induced infarction. have been shown to improve survival and morbidity [5 strokes of leMay et al. 19(11):1411-1419(1,988)]. AICA riboside and The professional driver of the present invention.

Because blood glucose can be lowered, it can reduce ischemic damage to the central nervous system (CNS). ) could be protected.

Another aspect of medical importance is that induced by elevated levels of homocysteine. treatment of symptoms or neurological disorders (eg, vitamin B12 deficiency).

AICA soboside prodrugs can be used for these purposes as well. Ru.

During a seizure, certain nerve cells become abnormally inflamed. ATP catabolism is an abnormal flame It greatly promotes adenosine production in firing cells. life is increasing. Adenosine has a pronounced anticonvulsant effect, so it has a significant effect on the brain. It is called a natural anticonvulsant.

Adenosine plays an important role as an inhibitory neuromodulator in the brain I think that the. This action of adenosine is apparently mediated by specific extracellular receptors. mediated. Adenosine has both postsynaptic and/presynaptic effects. Ru. Some of the well-established effects of adenosine on nerve tissue include inhibition of nerve inflammation. There is harm and inhibition of calcium-dependent neurotransmitter release. Adenosine and its Metabolically stable analogues have profound anticonvulsant effects behaviorally. It has anticonvulsant and sedative effects.

As mentioned above, adenosine has been proposed to serve as a natural anticonvulsant. and substances that modify its extracellular concentration may function as modulators of seizure activity. Ru. Additionally, adenosine is a neuromodulator. It acts as a powerful vasodilator and an inhibitor of granulocyte oxygen free radical production. It is an anti-arrhythmic agent.

Because adenosine has many actions, in practice it can be used in specific ways against specific pathogenic attacks. "Retaliation molecules" released to protect cells.

ry molecule) J.

However, to maintain effective extracellular therapeutic concentrations of adenosine in target organs, At concentrations that must be administered systemically to patients, adenosine is toxic and However, administration of adenosine alone has limited therapeutic applications. Furthermore, the body Most cells in the body have receptors for adenosine, so adenosine Techniques that increase syn concentration throughout the body are based on normal cellular physiology. This can lead to drastic changes that are not desirable.

Summary of the invention The present invention is characterized by increasing extracellular adenosine concentration in neuropathic tissues. To prevent or reduce neural tissue damage associated with neurodegenerative diseases in the above individuals. The purpose of this method is to

In one aspect of the invention, a therapeutically effective amount of an agent that increases extracellular azo/syn is administered to a patient to increase extracellular adenon-7 concentration. extracellular adhesion Suitable substances that increase nosine concentration include adenosine modulators (adenosine regulating agents) (e.g., regulating agents of adenosine catabolism) inhibitor and enhancer of adenosine production), inhibitor of adenosine transport -, and adenosine agonists.

Particularly preferred both groups of substances that function as adenosine modulators include AICA ribbons. or less per equivalent weight of AICA ribosyl moiety and AIcA ribosyl moiety. and one hydrocarbyloxycarbonyl or hydrocarbylcarbonyl moiety AICA ribonadobrodola, which consists of a modified AICA riboside having .

In another aspect, the present invention provides for increased release of excitatory amino acids (EAA). Increasing the extracellular concentration of adenosine prevents nerve tissue damage caused by The purpose is to

Adenosides such as AICA riboside and its prodrugs disclosed herein. In addition to exhibiting beneficial adenosine modulation/EAA inhibition abilities, the Site- and event-specific avoidance of undesirable global effects of adenosine agonists There is.

In a further aspect, the invention provides treatment for Parkinson's disease and related neurodegenerative diseases in patients. treatment by increasing the extracellular concentration of adenosine in the brain of the patient. The purpose is to do something. Such diseases include Alzheimer's disease, amyotrophic lateral sclerosis (ALS) and Huntington's disease.

Brief description of the drawing Figure 1 shows the effects of ketamine on the development of methamphetamine-induced parkinsonism. FIG.

Figure 2 shows the effects of ketamine on the development of methamphetamine-induced parkinsonism. 1 is a graph showing the effects of caffeine and CHA (in the presence and absence of caffeine).

Figure 3 shows that CH on MPTP-induced decrease in tyrosine hydroxylase activity. It is a graph showing the effect of A.

Figure 4 shows that CH on MPTP-induced decrease in tyrosine hydroxylase activity. 1 is a graph showing a comparison of the effects of A and MK-801.

Detailed description of the invention The method of the present invention provides an extracellular concentration of adenosine or an adenosine agonist in neural tissue. The purpose is to prevent or reduce nerve tissue damage by increasing the degree of injury. child One proposed mechanism for the protective effects of excitatory amino acids (EAA) Associated with inhibition of induced neurotoxicity. Excitotoxicity is methane in Parkinson's disease Related to the phetamine animal model [5science of 5onsalla et al. 243:398-400 (1989)]. According to the methamphetamine model, God The mechanism of degeneration involves the release of newly synthesized neurotoxic dopamine oxidation products. Output is involved. In other neurodegenerative disease models, methylphenyltetrahydropi Lysine (MPTP) is involved, and MPTP is 1-methyl-4-phenylpyrylysine. Causes damage by Dahlia conversion to Zinium ion (M　P　P　　　) That's what I think. The latter is then selectively taken up by dopaminergic neurons. A redox cycle that uses neuromelanin as an electrical source superoxide is produced in Mydophundria by [Markey et al. Medical Research Reviews 8(4):389 -429 (1988)].

The methods of the present invention demonstrate neurodegeneration in both methamphetamine and MPTP models. It has been shown to be effective in preventing the effects of

Preferred adenodinergic compounds One aspect of the invention provides that extracellular adenosine production is increased to the extent that it can protect against neurodegenerative disorders. Concerning the use of substances that increase As mentioned above, these materials contain ArCA Includes adenosine modulators such as fosides, and their analogs and prodrugs. It will be done.

Substances that increase extracellular azo/sin a level include inhibitors of adenosine production or are adenosine modulators, e.g. inhibitors of azone metabolism or adenosine production. There are raw enhancers, etc.

Substances that can inhibit cellular transport of adenosine include substances that specifically act in this way. , a substantial competitive inhibitor of adenosine uptake, and others that inhibit non-specifically There are substances. p-Nitrobenzylthioinosine and diviridamol are competitive inosine. Considered to be inhibitors, drugs such as colchicine, phenethyl alcohol, and pababerine A variety of other chemicals inhibit uptake non-specifically.

Furthermore, the extracellular concentration of azo/syn is a chemical that inhibits the enzymatic degradation of adenonone. It can also be increased by using quality. One group has a Azo/syndeaminase, an enzyme involved in converting denosine to inosine There are inhibitors of

As inhibitors of adenosine deaminase activity, copolmain, 2- Deoxycoformycin, and erythro 9-(2-hydroxy-3-nonyl) There is adenine hydrochloride.

Adenosine receptor agonists and antagonists include structural modifications to the purine ring. Adenosine analogs with decorations, adenosine analogs with modified substituents bonded to the purine ring There are analogs and analogs with modifications or alterations in the binding site of the carbohydrate moiety.

Inhibitors of adenosine transport, inhibitors and inhibitors of adenosine deaminase Denosine agonists increase extracellular adenosine more than adenosine alone. Although they offer the advantage of increasing , and the opposite, mainly due to its non-selective action on most cell types. It may present inconveniences such as side effects. Most cells in the body are adenosylated Because it has functional receptors for adenocarcinoma, Techniques that increase the concentration of syn [Purine Metabolism in Man, (D e　Baryni! , 5iffiIlonds and Mutter), Pre No 0 Breath, New 1 Yo 1 Ri (1984)]. In addition, adenosine deaminer Zein inhibitor (±, also prevents the degradation of deoxy7 adenone, which is a powerful immune system) [Gruber et al. Ann, New York Acad, Sci. 451:315-318 (1985)].

Therefore, extracellular Adenosinergic agents that selectively increase adenosine levels are also preferred. It is. Therefore, a preferred embodiment of the method of the present invention is to use extracellular adenosine in neural tissue. Preventing neurodegenerative disorders using adenosine modulators that selectively increase concentrations The purpose is to

by increasing the extracellular adenosine concentration (or one or more adenosine concentrations). Compounds that are thought to alter the biochemical pathways of cancer metabolism (e.g., adenosine AMP deaminase, AMP nucleotidase). such that the net result is an increase in the extracellular concentration of Azo/Syn. This is a thin one or more of the following: increased intravesicular production and/or decreased adenosine catabolism can be caused by the process of Compounds useful in the methods of the invention include, for example: , compounds broadly classified as purine nucleosides, and AICA riboside, AIC A ribotide, 1-β-D-ribofura/silu LB-1,2,4-triazole- Related analogs such as 3-carboxamide (ribavirin) and ribavirin-phosphate There are various pro-forms of these compounds. These compounds are taken up by cells and, optionally, the -phosphate form, and to a lesser extent the diphosphate form and the -phosphate form. It is thought to be converted to the triphosphate form. Additionally, this includes the following compounds: (1) Endogenization of metabolites such as AICA riboside or purine intermediate metabolites Substances that can increase the synthesis or compounds that can form these metabolites, e.g. Nylaminoimidazole carboxamide (SAICA) riboside, (2) AI Substances capable of assembling CA-riboside or its metabolites, e.g. Rexate, and (3) increase the volume of AICA riboside in the bacterial flora. substances such as sulfonamides. These compounds may be used prophylactically in some cases. and/or to another patient in direct response to physical condition. . Extracellular concentrations of purine nucleosides or adenosine and/or adenosine analogs Analogs that increase the concentration range from 0.105 mmolar to 0.5 mmolar. In the case of A, IOA riboside, it can be administered to biological systems over multiple times. Usually administered at concentrations up to 0.5 mmolar.

AICA riboside and other adenosine regulators without the negative effects of adenosine. Novel compounds exhibit positive biological effects and, in some cases, ameliorated properties. The Prince Prodrug/Analog and Analog of Continued patent application rArcAIJ PosiF (+Drag JJ (USSN 301. 222), concurrent patent application rAIcAIJ Bosido filed on September 15, 1989 Compounds and Methods for Lowering Supply and Blood Glucose J (USSN 408,107 ), and USSN 466.979 rAIcA Ri filed on January 18, 1990. ``Compounds and Methods for Lowering Boside Supply and Blood Glucose'' (this (incorporated herein by reference). The compounds described herein are Can be used as a prodrug. These new compounds are typically: Demonstrates improvement over one or more of the AICA ribosides described: 1) Stronger Adenosine regulatory effect, 2) prolongation of half-life, 3) increase in brain penetrating capacity, 4) oral/s j) increased ioavailability, 5) increased myocardial targeting ability, 6) in some cases, Synergistic action with AICA riboside itself. AICA riboside prodrugs of the present invention It can be used in any method.

Adenosine or inosine may cause stroke, heart attack or seizure activity such as angina, arrhythmia. or rapid cellular energy utilization during symptoms resulting from reduced blood flow (ischemia), etc. It is produced from adenosine triphosphate by Normally, when such an event occurs, the Nosine production is greater than adenosine production. Low blood flow area during coronary vessel occlusion For example, the ratio of inosine to adenosine is about 100:1. Next (A Then, a certain percentage of inosine and adenonine is excreted from each cell, They exist directly in the extracellular environment. The above adenosine modulators are described herein. It is useful in the method described above to increase the extracellular concentration of this adenosine and even to It has been shown to reduce the production of syn to certain settings. Variations in azo/syn production occurs only in the area of and concurrently with net ATP use, and Because denosine is rapidly degraded, azo/sine levels are does not change significantly. Therefore, these adenosine modulating substances according to the method of the present invention are Rather than increasing systemic, i.e. global, adenosine, localized levels of extracellular adenosine This will increase the amount locally.

The purine nucleoside analog AICA riboside is metabolized to uric acid; substances such as allopurinol or other drugs that inhibit uric acid synthesis, or brobenecid. Which uric acid excretor should be used with?

Methotrexate and its metabolites, such as ribavirin, are AICA Ribondo!・ The specific substance that inhibits transformylase is the endogenously synthesized AICA ribosome. cid and may have similar effects as administering purine nucleosides. . AICA riboside or AlCA riboside and AXCA riboside transformira co-administration with inhibitors of enzymes should show at least an additive effect. difference In addition, any intermediate of de novo purine nucleotide synthesis (purine synthesis (after the first relevant step for AIC) or their nucleosides or bases It can be expected to be rapidly converted to A riboside. For example, 5AICA boside or its nucleotide or base.

When contacted with cells, adenosine-regulated purine nucleosides and useful in the methods of the invention The analogue enters the cell, where it is cleaved, if necessary, by adenosine kinase. or if a base is administered, it can be phosphoribosyltransformed. It is converted into nucleotides by the enzyme chelase and becomes purine nucleotide-phosphate. It is thought that this may eventually become a nucleoside diphosphate or triphosphate.

These diphosphate or triphosphate forms constitute the pool for decomposition into the -phosphate form. There are cases where

In one aspect of the invention, preferred AIs include compounds of the formula: Using a CA riboside prodrug compound: [where xl, islet and X are individually (a) Hydrogen, Q 0 (b) -CR, or -00RI (wherein R1 is individually a hydrocarbon having preferably 1 to about 24 carbon atoms) 24 carbon atoms, individually mono- or mono- or di-hydrocarbyl amines The force A, which is that of the number), and two of +1 (c) X, , X- and X, are Together, they form a cyclic carbonate group.

Preferred R and R groups include lower alkyl groups. Among the lower alkyl groups One preferred group of is all those containing at least one secondary or tertiary carbon atom. It is something that Another preferred class of lower alkyl groups is up to about 6 carbon atoms. It has the following. Hydrocarbyl groups having more than 24 carbon atoms can also be used.

Some preferred compounds contain one or two ester groups. Special Preferably, the ester is present at the 3' or 5' position or both positions of the ribosyl ring. It is a compound that has a group.

For multiple indications, transropaioavail may offer therapeutic benefits. It is convenient to orally administer these prodrugs that exhibit virility. preferable. Therefore, one or more of X, X, and X3 are short chain hydrogen atoms. Preferred are prodola knobs containing locarbyl carbonyl groups. Xl is small Nobutyryl or prodola, which is pivaloyl and where X and X are both hydrogen. Knob, and X,,X. and X, both of which are acetyl, are liquid or can be administered orally in either solid (e.g., capsule) form with increased bioavailability. It is a particularly preferred compound because it shows availability. Furthermore, XI is n-butyryl, and X and X are both hydrogen, and Also preferred are prodrugs in which both and X are acetyl and X is hydrogen. Special Preferred compounds are certain prodrug compounds that are isolated in useful crystalline forms. Specifically, 2°, 3', 5'-triacetyl AICA riboside 3'.

5°-diacetyl AICA riboside and 3′-neopentoxycarbonyl . Furthermore, in acetyl-substituted prodrug compounds, the leaving group is acetate. This is advantageous because it is relatively pharmacologically inactive.

(Margin below) Preparation of ArCA riboside prodrug compound AICA riboside prodrug compound The product can be produced according to the following reaction formula. In formula E, X3, X2, Xl, R4 and R1 are the same as defined in formula (I) above. be] Reaction (1) involves compound (II), AICA riboside and compound (I[[), appropriate This is carried out by mixing acid chlorides, acid anhydrides or chloroformates in a solvent. Acid Lorides are routinely prepared by conventional operations such as reacting the corresponding acid with thionyl chloride. It can be manufactured by the method. Acid chlorides and acid anhydrides are commercially available. There is also one. Although many chloroformates are commercially available, phosgene can be It can be manufactured in a conventional manner by conventional operations known to those skilled in the art, such as reacting with cole. You can also Reaction (1) is carried out at about -10°C to about 5°C, preferably about -5°C. to about 0° C. and is generally completed in about 2 to about 4 hours. Easy to operate To do this, the reaction is carried out in a solvent. Suitable solvents include dimethylformamide (D MF), pyridine, methylene chloride, etc. For convenience, the reaction was carried out at atmospheric pressure. Do it with The reaction product(s) can be processed by conventional methods such as column chromatography, crystallization, etc. more isolated. This reaction allows the 2'-13'- and/or 5- It is possible to obtain a mixed product of mono-, di- and tri-esters at the ° position. be understood. The resulting ester product was analyzed by thin layer chromatography (TLC). , high pressure liquid chromatography (HPLC), column chromatography, crystallization The separation can be carried out by conventional methods well known to those skilled in the art, such as chemical reaction.

The 5'-monoester yields an intermediate protected at the 2' and 3° positions. It can be produced by a conventional method according to the following reaction formula: (Margin below) agent] Reaction (2) involves mixing and condensing compounds (II), (IV), (V) and (VT). do. These reactants may be mixed in any order, but compound (n) It may be convenient to add it to a mixture of (IV), (V) and (Vr). this The reaction is carried out at a temperature of about 10°C to about 25°C, preferably about 15°C to about 25°C. The reaction is generally completed within about 45 minutes. The intermediate (Vr) is prepared in the usual manner. Therefore, it is isolated.

Reaction (3) involves the reaction between intermediate (■) and a suitable acid chloride, acid anhydride or chloroform. The reaction is carried out as described for reaction (1).

Reaction (4) optionally involves removing cyclic protecting groups from the 2′ and 3′ positions. It is a process. This is carried out by reaction of compound (IX) with a suitable deprotecting agent.

Suitable deprotecting agents include, for example, water/acetone, tetraethyl-ammonium fluoride, Ride/TT (F, H resin in acetic acid/water, formic acid/water. Such deprotection The reaction is conventional and well known to those skilled in the art.

The mixed ester compound is prepared by first converting AICA riboside into a suitable acid salt according to reaction (1). React with loride or acid anhydride to add acyl ester group, then the obtained React the acyl ester-substituted compound with the appropriate roloformate according to reaction (1). It can be produced by a conventional method by subjecting the compound to a mixed ester. a Alternatively, the mixed ester compound is first subjected to AICA according to reaction (1) and reaction (2). The riboside is converted into a monoacyl ester, and then according to reaction (1), the obtained by reacting the purified monoafluorated product with the appropriate roloformate. can also be manufactured. Furthermore, certain mixed esters can be used for reaction (1) or According to (2), ArCA riboside is first converted to mono-alfky 7 carbonate form. Then, the purified carbonate ester obtained was treated with an appropriate acid according to reaction (1). Manufactured by reaction with chloride or acid anhydride.

Purpose The method of the present invention uses a substance that increases extracellular adenosine concentration to treat neurodegenerative diseases. related to treating disease. As previously mentioned, AICA riboside and herein Adenone modulators, such as the AICA riboside prodola knob compounds described, are Useful to treat conditions where increasing extracellular concentrations of denosine would be beneficial It is.

The studies described herein induce neurodegenerative and parkinsonian symptoms in animals. Two substances were used: MPTP and methamphetamine. Neurodegeneration is a male 20xg/kg of MTP or 5mg for Swiss Webster mice The animals were induced by systemic injections of methamphetamine/kg every 2 hours.

CHA was injected 3 times at 0.5 mg/- 0.3 and 6 hours after the first toxin injection. CHA was administered by. Ketamine is 100mg/each for each toxin injection. kg were administered simultaneously. Animals were allowed to live for 7 days before being killed, at which point the brain was removed. The striatum was removed. The tissue was homogenized and the method of Re1nhard et al. [According to Law CLffe 5 Sciences 39:2185-2189] were analyzed for hydroxylase activity.

Both methamphetamine and MPTP are associated with striatal tyrosine hydroxylase activity. decreased by 40-45%. CHA (p<,01) or ketamine (glutamine ceptor antagonist) (p<,05), it It was effective in completely reversing amphetamine-induced decline. The inventors Although not tested, Sonsal says ketamine is not effective against MPTP. It has been reported by la et al. On the other hand, CHA also decreases in this model. prevented (p<,01). Both models utilize the protection provided by CHA. Concomitant administration of caffeine (30-50xg/kg), an adenosine antagonist We were able to partially reverse the situation by applying this method.

CHA predicts the development of MPTP and methamphetamine-induced parkinsonism. Prevented. This is because in this methamphetamine model, tyrosine hydroxyl It has only been shown that it is significantly effective by discriminating the enzyme. This is a better protection than reported for EAA blockers, which are not shown.

Figure 4 shows the inefficiency of NMDA-specific EAA receptor blocker MK-801. It is shown in

Furthermore, depending on the CHA dose required, it may not be similar to FAA Pro7car; Does not exhibit severe sedative effects. This protective effect is due to adenosine receptor antagonists. It is adenosine receptor-mediated because it can be reversed by a strike.

This data suggests that adenosine is an endogenous neuroprotective agent in Parkinson's neurodegeneration. The results suggest that adenosinergic It shows the possibility of using the plan.

The present inventors demonstrated that adenomatosis can be used to treat experimentally induced Parkinson's disease in animals. One aspect of the invention is that syn receptor agonists can provide significant protection. I found it. The disease progresses over a long period of time (many years) and is generally mild. It is a disease that can only be recognized by tremors. Current treatment methods are symptomatic (L-DOPA), but does not stop the progression of the disease. stomach. The method of the invention acts through the adenosinic mechanism (adenosinergic). Prevention of this progressive deterioration in Parkinson's disease patients is based on the use of substances. I can do it. The term "adenosinergic substance" refers to one of the following substances or Means the combination: 1) adenosine receptor agonist, 2) adenosine receptor agonist; syn transport inhibitor, or 3) an activator that increases extracellular adenosine concentration. enzyme inhibitor. One or a combination of these substances may be It can be administered chronically in the vehicle position. The effects of substances in category 2 or 3 above are relatively sensitive and their side effect profile is relatively manageable, so chronic These substances are probably the most used when performing long-term treatments. available.

Cyclohexyladenosine (CHA), an adenosine receptor agonist, , MPTP and methamphetamine model for the development of parkinsonian syndrome in rats. It has been shown to inhibit In response to MPTP or methamphetamine, the striatum ( Dopamine-rich areas of the limbic system that are selectively degraded in Parkinson's disease ) The decrease in tyrosine hydroxylase was monitored. MPTP and meth In both models of etamine, CHA administration was shown to confer significant protection. [Examples 1 to 4 and FIGS. 1 to 4].

The adenosinergic compound useful in the methods of the invention is about 0.011 g/kg/day. Doses from about 500 to about 500 g/kg/day are effective, preferably from about 15 to about 15 g/kg g/day to about 200 mg/kg/day. This dosage range is Precautions to prevent neurodegenerative disorders associated with undesirable limited or reduced direct current It is suitable when the compounds useful in the present invention are used as drug preventive agents. preventive medicine and When used with a temperature of at least about 0. lxg/kg/day A I-CA Libono or ArCA ribotide, preferably about 1. Ozg/kg/day to about 500zg /lcg 7 days, and more preferably about 20 mg/kg/day to about 1°O+g/k g/day is considered. For some adenosinergic compounds, the blood-brain barrier Doses of 200-500a+g/kg/day or more may be required for . However, this can be done at lower doses using brain-directed prodrugs. Wear.

These azo/sinergic compounds (e.g. prodrugs of AICA ribotide) are This is because it is not easily broken down by the body's extracellular enzymes or by exposure to the low pH in the stomach. Oral administration is the most common way to deliver these compounds to patients. Furthermore, these compounds For intravenous administration, direct intramuscular injection, subcutaneous administration, topical administration to the skin or mucous membranes, rectal administration, or inhalation. It can also be administered by injection. Compositions suitable for pharmaceutical use are well known. It is. prodrugs, i.e., the activity of adenosinergic substances when introduced into the body; Those that are metabolized according to sex type can also be used.

In order to facilitate understanding of the present invention, examples are provided below, including a series of experimental results. Ru. The following examples relating to the present invention are illustrative and should not be construed as limiting the invention. Of course, this should not be the case. Furthermore, it is considered to be within the knowledge of a person skilled in the art, Modifications of the invention, now unknown or later developed, may also be included in the claims set forth below. should be considered to be included within the scope of the requirements.

Example 1 Male Swiss Webster mice were treated with 5 mg/kg (+) methamphetamine for 2 days. Striatal reduction was induced by systemic administration four times at intervals. CIA x g/kg dose 3 times, i.e. 15 minutes before the first methamphetamine injection; The other two doses were administered 3 and 6 hours later. Ketamine is lOOzg/kg Four doses were administered. This injection is prescribed as a 09% saline injection. This was done at the same time as the tamphetamine injection. All of these compounds were tested in a saline vehicle. It was delivered as an intraperitoneal porous at a dose of 1 x Q/100 g animal body weight.

After 7 days, the animals were sacrificed and the brains were quickly frozen on dry ice. Then 50mM The brain was thawed in potassium phosphate buffer (pH 6,1>) and the striatum was excised. Tissues from the two hemispheres of the garment were combined and contained 0°2% Triton X-100. Homogenize in 100 μQ of the same buffer as above and 10 minutes at 17,0 OOXG. Centrifuged. 1. F.

The method of Reinhard et al. [Life 5 Sciences 39:2185 (1 986)], 6,7-dimethyltetrahydropterin was used as a cofactor. Tyrosine hydroxide of the obtained supernatant in 5011M phosphate buffer (pH 6.1) containing Roxylase activity was analyzed.

The results obtained are shown in FIG.

Example 2 Striatal reduction was induced according to Bucotocol as described in Example 1.

Again, ketamine was administered 4 times to methamphetamine and 1001 g/kg each. However, in this experiment, the saline injection was 0.3 and 6. After an hour, this is similar to the administration of other drugs. CHA Ha, 1. wg /kg, 0.5xg/hire, Ii30xg/kg 0,5 with added caffeine It was administered as three injections at either xg/ksx.

After 7 days, the animals were sacrificed and the striatum was excised. The striatum obtained from the two hemispheres of winter clothes was 501IIM phosphorus containing 0.2% Triton X-100. Homogenized in 1 volume of potassium acid buffer. The obtained homogenate was ．． ○ Spin at 0OXG and transfer the supernatant to tyrosine hydroxyl as described in Figure 1. were analyzed for lase activity.

The results obtained are shown in FIG.

Implementation PI3 In inducing Parkinson's syndrome in mice using MPTP, The dose of 20*g/kg was administered systemically 4 times every 2 hours.

CHA at 0.5xg/kg injection or added with cuffin at 50wg/kg injection. What you got is 0. 0.3 and 6 after the first MPTP injection as an injection of 9% saline. The drug was administered three times in total. All injections were performed at a dose of 1 x Q/100 g of animal body weight. It was given as an intraperitoneal porous injection.

After 7 days, the animals were sacrificed and the brains were removed. The striatum was excised and obtained from the two hemispheres. The ingredients were collected, weighed, and added to 50 nM potassium phosphate buffer (pH 6, IN). Homogenized in volume. The resulting homogenate was then treated as described in Example 1. Tyrosine hydroxylase activity was analyzed in this manner.

The results obtained are shown in FIG.

Example 4 Parkinsonism was induced in mice using MPTP as described in Example 3. induced. (: HA (5mg/kg) and MK-801 (3,5zg /kg) in 0.9% saline and O13 and 6 of MPTP injection of rjlW. Three doses were given after hours. All these injections were performed at a dose of 1112/100 g d. It was given as an intraperitoneal porous injection of body weight.

Tyrosine hydroxylase activity was analyzed as described in Example 3.

The results obtained are shown in FIG.

F/G, 4゜ MPTP 9E:’JL international search report

Claims (26)

[Claims] 1. Excitotoxicity caused by or caused by neurodegenerative conditions excitotoxicity, resulting from increased release of excitatory amino acids in affected animals. A method for preventing neural tissue damage caused by A method characterized by increasing the extracellular concentration of adenosine in its surroundings. 2. Treatment with a substance that increases the extracellular concentration of adenosine in or around the neural tissue Claim 1, wherein adenosine concentration is increased by administering to an individual a scientifically effective amount. The method described in. 3. the substance is an adenosine modulator, an adenosine agonist, or an adenosine transporter; 3. The method according to claim 2, wherein the method is an inhibitor. 4. 4. The method according to claim 3, wherein said substance is an adenosine agonist. 5. 4. The method according to claim 3, wherein the substance is an adenosine modulator. 6. 3. The method according to claim 2, wherein the neural tissue is the brain or spinal cord. 7. Claim 6 wherein said excitotoxicity is caused by or caused by brain trauma. The method described in. 8. Neurodegenerative conditions include Birkinson's disease, Alzheimer's disease, amyotrophic lateral sclerosis, or Huntenton's disease, the method according to claim 2. 9. the substance is an adenosine modulator, an adenosine agonist, or an adenosine transporter; 9. The method according to claim 8, wherein the method is an inhibitor. 10. in a manner that reduces neural tissue damage associated with neurodegenerative diseases in affected animals. characterized by increasing the extracellular concentration of adenosine in the neurological disorder. How to do it. 11. administering to the individual a therapeutically effective amount of a substance that increases adenosine concentration; 11. The method of claim 10, wherein the adenosine concentration is increased by: 12. The substance that increases the adenosine level is an adenosine regulator, adenosine 12. The method of claim 11, wherein the method is an agonist or an adenosine transport inhibitor. 13. Claim 12 wherein the substance that increases adenosine concentration is an adenosine regulating substance. The method described in. 14. The adenosine modulator is a purine nucleoside or an analog or protein thereof. 14. The method according to claim 13, wherein the method is a drug. 15. The adenosine modulating substance is AICA riboside or AICA riboside prodrug. 15. The method according to claim 14, wherein the method is a program or an analog. 16. Claim wherein the substance that increases adenosine concentration is an adenosine agonist. 12. The method described in 12. 17. The neurodegenerative diseases include Birkinson's disease, Alzheimer's disease, and amyotrophic lateral sclerosis. 13. The method according to claim 12, wherein the method is Huntington's disease. 18. A method for preventing neurological tissue damage associated with neurodegenerative conditions, the method comprising: Prophylactic administration of a therapeutically effective amount of a substance that increases the extracellular concentration of denosine A method characterized by: 19. 19. The method of claim 18, wherein the neurodegenerative condition is Parkinson's disease. 20. If the substance is an adenosine modulator, an adenosine agonist, or an adenosine transporter, 19. The method of claim 18, wherein the method is a transport inhibitor. 21. 21. The method of claim 20, wherein the substance is an adenosine agonist. 22. 21. The method according to claim 20, wherein the substance is an adenosine modulator. 23. The neurodegenerative diseases include Birkinson's disease, Alzheimer's disease, and amyotrophic lateral sclerosis. 21. The method according to claim 20, wherein the method is Huntington's disease. 24. Progressive brain deterioration associated with neurodegenerative diseases in animals affected by the disease A method for reducing the extracellular concentration of adenosine, including the treatment of substances that increase the extracellular concentration of adenosine. A method comprising administering to said animal a therapeutically effective amount. 25. If the substance is an adenosine modulator, an adenosine agonist, or an adenosine transporter, 25. The method of claim 24, wherein the method is a transport inhibitor. 26. The neurodegenerative diseases include Birkinson's disease, Alzheimer's disease, and amyotrophic lateral sclerosis. 26. The method according to claim 25, wherein the method is Huntington's disease.