UA72736C2 - N-6-substituted 7-deazapurines (variants), method for synthesis, pharmaceutical composition for treating adenosine-mediated disorders (variants) - Google Patents

Abstract

Novel deazapurines of the formula: are disclosed which are useful for the treatment of adenosine receptor stimulated diseases.

Description

Description of the invention

Adenosine is a ubiquitous modulator of many physiological processes, especially in the 2 cardiovascular and nervous systems. The action of adenosine is apparently mediated by specific receptor proteins on the cell surface. Adenosine modulators differ in their physiological functions, including such functions as inducing a sedative effect, vasodilation, reducing heart rate, inhibiting platelet aggregation, stimulating gluconeogenesis, and inhibiting lipolysis. In addition, it was shown that, in addition to its effect on adenylate cyclase, adenosine opens potassium channels, reduces the flow of 70 through calcium channels and inhibits or stimulates the metabolism of phosphoinositidine under the action of receptor-mediated mechanisms I (see, for example, "S.E. MyPeg 5 V.biweip "Adepozipe Keseriog Apiadopiviv: Zygisijgez ap Roiiepiia!

Tpegaretsiis Arriisayopve", Sysptepi RPpagtasetsiisai UOevidp, 2:501 (1998), and S.E. Miyeg "AgAdepozipe Keseriog

Apiadopiv: Ehr. Orip. Tpeg. Rayepiv 7(5):419 (1997)).

Adenosine receptors belong to the superfamily of purine receptors, which are currently divided into 12 P receptors (adenosine) and P receptors. (ATP, AOR and other nucleotides). To date, four subtypes of nucleoside adenosine receptors have been cloned from different species, including humans. Receptors of two subtypes (A; and Aga) have affinities for adenosine in the nanomolar range, and receptors of the other two known subtypes A with Az are low-affinity receptors with an affinity for adenosine in the low micromolar range. Activation of adenosine receptors A 5 and Az can lead to inhibition of adenylate cyclase activity, while activation of Aza and Az leads to stimulation of adenylate cyclase.

Several A-receptor antagonists have been developed for the treatment of cognitive impairment, renal failure, and cardiac arrhythmia. It has been suggested that A 24 receptor antagonists may be used to treat patients suffering from Parkinson's disease (Parkinson's disease).

In particular, it is likely that with local delivery, antagonists of adenosine receptors may be effective for the treatment of allergic inflammatory reactions and asthma. Data (eg, "Muse 85 Meigde OMA (39 apiizepze Tpegaru og Avijta ip ap Apita! Mode!" Maitsge (1997) 385:721-5) indicate that under these pathophysiological conditions, A- antagonists can block the contraction of smooth m' lower respiratory tract epithelium, while antagonists of A2B5 or Az receptors can block mast cell degranulation, reducing the release of histamine and other inflammatory mediators.A 25 ee receptors have been found to be present throughout the gastrointestinal tract, especially in the epithelium of the large and small intestine. It has been suggested that A55 receptors mediate the cCAMP response of Zygopterygii.

U. Vioi. Spet., (1995) 270:2387-94). -

Adenosine receptors have been shown to be present in the retina of many mammalian species, including cows, pigs, monkeys, rats, guinea pigs, mice, rabbits, and humans.

Rivogei Oivigirtsiop oi Adepovipe Keseriogz ip Mattaiap Keiippa, doigpa! oi Meyhospetivigu, moishte 54, radevz v 648-655 (1990); MUsodz ei ai.,, SPagasiegizayop oi Adepozipe Ag Keseriog Vipdipd 5Zye ip Vomipe Kepayi

Metargapez, Echretitopia!I Eue Kezeagsp, moYishTe 53, radez 325-331 (1991); and Vgaaz ei aI., Epdodepoiz adepovipe apa adepozipe geserioge Iosarylei (o dapadip seiv oi (She gaypa, Rgoseedipudz oi (She Mayopa! Asadetu oi "

Zsiepse, myoshte 84, radez 3906-3910 (1987)). Recently, the work of U/Shatve reported the observation of Z sites of adenosine transport in a cultured cell line of the human retina (MMPShiatve ei ai., Mysieovide s Tgapzrogi Zyez ip a Siyigei Nytap Keiyipa! SeiYi Iipe Eviarizyei ru 5M-40 F Apidep Sepe, Sitepi Eue

From" Kezeagsi, myoshte 13, radez 109-118 (1994)|.

It has previously been suggested that compounds that regulate adenosine uptake may be used as potential therapeutic agents for the treatment of retinal and optic disc lesions. U.S. Patent No. 49, No. 5,780,450, to Zpade, discusses the use of adenosine uptake inhibitors for the treatment of eye diseases. The Zpade patent does not describe the use of specific Az receptor inhibitors. The full description of US Patent No. 5,780,450 is incorporated herein by reference.

The need to obtain other antagonists of adenosine receptors as pharmacological agents 7 remains relevant, and drugs for the treatment of the above-mentioned are of particular interest

Fa 020 diseases and/or pathological conditions.

The present invention is based, at least in part, on the discovery that the M-6-substituted 7-deazapurines described below can be used to treat conditions responsive to M-6-substituted 7-deazapurine.

Examples of such conditions are conditions associated with increased activity of adenosine receptors, such as bronchitis, gastrointestinal diseases or asthma. These conditions differ in that activation of the 29 adenosine receptor can lead to inhibition or stimulation of adenosine cyclase activity. Compositions

GF) and the methods of this invention include enantiomerically or diastereomerically pure M-6b-substituted 7-deazapurines. Preferred M-6b-substituted 7-deazapurines are those M-b-substituted 7-deazapurines that have an acetamide, carboxamide, substituted cyclohexyl, for example, cyclohexanol or urea group linked to

M-6-nitrogen with the help of an alkylene chain. The present invention relates to methods of modulating adenosine receptor(s) in a mammal by administering to the mammal a therapeutically effective amount of M-6b-substituted 7-deazapurine such that this administration results in modulation of adenosine receptor activity. Suitable adenosine receptors are receptors of the Az, Az or Az families. In a preferred embodiment of the present invention, M-6-substituted 7-deazapurine is an adenosine receptor antagonist.

In addition, the present invention relates to methods of treating diseases that respond to M-6-substituted 7-deazapurine, for example, asthma, bronchitis, allergic rhinitis, chronic obstructive pulmonary disease,

renal disease, gastrointestinal disease and eye disease in a mammal by administering to said mammal a therapeutically effective amount of M-6-substituted 7-deazapurine such that said administration results in treatment of said mammal. Corresponding M-b6-substituted 7-deazapurines are compounds having the general formula I:

In zhk with od and their pharmaceutically acceptable salts. Each of C and C independently represents a hydrogen atom or a substituted or unsubstituted alkyl, aryl or alkylaryl group, or taken together they form a substituted or unsubstituted heterocyclic ring. Kz represents a hydrogen atom or a substituted or unsubstituted alkyl, aryl or alkylaryl group. K,. represents a hydrogen atom or a substituted or unsubstituted alkyl, aryl or alkylaryl group. Each of K5 and Co independently represents a halogen atom, for example, a chlorine, fluorine or bromine atom, a hydrogen atom or a substituted or unsubstituted alkyl, aryl or alkylaryl group; or, K and K5 or Kv and Kv taken together will form a substituted or unsubstituted heterocyclic or carbocyclic ring.

In some embodiments of the present invention, each of K. and K" can independently represent substituted or unsubstituted cycloalkyl or heteroarylalkyl groups.

In another embodiment of the invention, K c is a hydrogen atom or a substituted or unsubstituted c heteroaryl group. In yet another embodiment of the present invention, each of K/, K5 and Ke can independently represent heteroaryl groups. In a preferred embodiment of the invention, C is a hydrogen atom, C is cyclohexanol, for example, trans-cyclohexanol, C is phenyl, C is a hydrogen atom, C is a metal group, and C is a methyl group. In another variant of the implementation of this invention, K. represents a hydrogen atom, Ko represents

Ez represents phenyl, K represents a hydrogen atom, and Kb and K5 represent a methyl group.

The present invention also relates to pharmaceutical compositions for the treatment of diseases in mammals that respond to M-6b-substituted 7-deazapurine, for example, asthma, bronchitis, allergic rhinitis, chronic obstructive pulmonary disease, renal diseases, gastrointestinal diseases and eye diseases This pharmaceutical composition includes a therapeutically effective amount of M-b-substituted 7-deazapurine and a pharmaceutically acceptable carrier. "

The present invention also relates to pharmaceutical compositions for the treatment of diseases corresponding to

M-6-substituted 7-deazapurine, in mammals. This packaged pharmaceutical composition includes a container containing a therapeutically effective amount of at least one M-6-substituted 7-deazapurine and instructions for the administration of said M-6-substituted 7-deazapurine used to treat a condition corresponding to "» on M-6-substituted 7-deazapurine, in a mammal.

The present invention, in addition, relates to compounds of formula I, where:

Cu is hydrogen; - and Co is a substituted or unsubstituted cycloalkyl, substituted or unsubstituted alkyl, or K 4 and K», taken together, form a substituted or unsubstituted heterocyclic ring;

Ez is substituted or unsubstituted aryl; -| KE. is hydrogen; and c 50 each of K" and Co independently represents hydrogen or alkyl, and their pharmaceutically acceptable salts. Deazapurins of this variant of the invention can be mainly selective antagonists of the Az receptor. These compounds 4) can be used for various therapeutic purposes, such as, for example, the treatment of asthma, renal insufficiency associated with heart failure, and glaucoma. In a particularly preferred version of the present invention, deazapurine is a water-soluble prodrug, which, due to the metabolism of ip mimo, for example, due to hydrolysis catalyzed by esterase, is able to be transformed into an active medicinal substance.

In another variant of its implementation, the present invention relates to a method of inhibiting the activity of an adenosine receptor (for example, Az) in a cell, which involves contacting this cell with M-6-substituted ime) 7-deazapurine (for example, preferably, with an adenosine receptor antagonist) .

In another aspect, the present invention relates to a method of treating eye damage in an animal (eg, 60 human) by administering to said animal an effective amount of M-6b-substituted 7-deazapurine of the formula |.

The preferred M-6b-substituted 7-deazapurine is an antagonist of adenosine receptors Az in the cells of this animal. The indicated eye lesion is a retinal or optic disc lesion, and this lesion may be acute or chronic. This lesion can occur as a result of glaucoma, edema, ischemia, hypoxia, or trauma. 65 The present invention also relates to a pharmaceutical composition containing M-6-substituted 7-deazapurine of formula I. This pharmaceutical preparation is preferably an ophthalmic ready-to-use preparation form

(for example, a ready-made preparation form for injection into the periocular, retrobulbar or intraocular area, a ready-made preparation form for systemic administration or a surgical solution for washing).

In another embodiment, the present invention relates to a deazapurine having the formula II:

You will find many of them

IS

(Fo where: X is M or SC; each of Cu and Co is independently hydrogen or substituted or unsubstituted alkoxy, aminoalkyl, alkyl, /5 aryl or alkylaryl, or taken together to form a substituted or unsubstituted heterocyclic ring, provided that K. and Co are both not hydrogen;

Ez is substituted or unsubstituted alkyl, arylalkyl or aryl;

KE. is hydrogen or substituted or unsubstituted C-Ceoalkyl;

Y represents hydrogen, substituted or unsubstituted alkyl, or K,; and I, taken together, form a substituted or 2o-unsubstituted heterocyclic or carbocyclic ring;

Ke is hydrogen, substituted or unsubstituted alkyl or halogen;

O is CH", O, Z or MK), where K; represents hydrogen or substituted or unsubstituted S.-Svalkil and

MU is unsubstituted or substituted alkyl, cycloalkyl, aryl, arylalkyl, biaryl, heteroaryl, substituted carbonyl, substituted thiocarbonyl or substituted sulfonyl; with the condition that if Kz is pyrrolidino, then K 4 is not methyl. The present invention also relates to pharmaceutically acceptable salts and prodrugs of the compounds of the present invention. and)

In a preferred embodiment of the invention, X represents SK 5, and O represents CH", 0, 5 or MH in formula II, where Ko is defined above.

In another variant of the formula II, X is M. Hezo In addition, the present invention relates to a method of inhibiting the activity of an adenosine receptor (for example, adenosine receptor Azs) in a cell by contacting this cell with a compound of the present invention. at

A preferred compound is an antagonist of this receptor. uh-

The present invention also relates to a method of treating gastrointestinal disease (e.g., diarrhea) or respiratory disease (e.g., allergic rhinitis, chronic obstructive pulmonary disease) in an animal with

335 By administering to said animal an effective amount of a compound of formula II (for example, an antagonist of the A zu receptor). uh-

The dominant mammal is man.

Distinctive features and other characteristic features of this invention will be detailed in the following description and in the claims. At the same time, it should be noted that specific variants of this invention are presented only for illustrative purposes and do not limit this invention. The fundamental differences of this invention can be " 70 Used in various variants of its implementation, which do not go beyond the scope of the invention. The present invention relates to methods of treating a condition responsive to M-6b-substituted 7-deazapurine in a mammal.

These methods involve administering to that mammal a therapeutically effective amount of M-6-substituted;" 7-deazapurine, described below, such that this administration results in the treatment of said condition in a mammal responsive to M-6-substituted 7-deazapurine.

The expression "condition responsive to the action of M-6b-substituted 7-deazapurine" means a disease or disease state characterized by being amenable to treatment with an M-6-substituted 7-deazapurine of the present invention described below, for example, treatment , which leads to a significant weakening of at least one symptom or the painful effect of this condition, which is achieved using the M-6b-substituted 7-deazapurine of this -I invention. Typically, such conditions are associated with an increase in adenosine levels in the host, which often results in the host developing physiological symptoms, including, but not limited to, toxin release, inflammation, coma, water retention, weight gain or loss, pancreatitis, emphysema

Rheumatoid arthritis, osteoarthritis, multiple organ failure, respiratory distress syndrome in children and adults, allergic rhinitis, chronic obstructive pulmonary disease, eye damage, gastrointestinal diseases, stimulation of skin tumors, immunodeficiency and asthma. |See, for example, "S.E.MyPeg 85 V.5ieip dv Adepovipe Keseriog Apiadopiviv: Zigosiigev apa Roiyepia! TPpegaretsshis Arriisanyope", Sipepi Rvagtaseciisa!

Oezidp, 2:501 (1996) and "S.E. MyPeg AhMAdepovzipe Keseriog Apiadopiv: Ehr. Orip. Tpeg. Raiepiv 7(5): 419 (1997) and

F) Y. ReoKiiziom, K.. RoIosa, "5.T. NoYidaie 5 I. Viaddiopi Adepozipe Aov geserioge: a pome! (pegaretsiis (agdei ip ka aziyta?" TIRBZ 19; 148 (1998)). Pathological effects that are often associated with the indicated symptoms, including, but not limited to, fever, rapid breathing, nausea, diarrhea, weakness, headache, and even death. -deazapurine, there are such painful conditions that are mediated by the stimulation of adenosine receptors, for example, A 4, Aha, Ag, Az, etc., as a result of which modulation of calcium concentration in cells and/or activation of RIS (phospholipase C) occurs. In an embodiment of the invention, the M-6b-substituted 7-deazapurine-responsive state is associated with adenosine receptor(s), for example, when M-6-substituted 7-deazapurine acts as an antagonist. corresponding positively reacting states, which can be cured by the compounds of this invention, are Art Anas, for example, are associated with adenosine receptor subtypes that mediate biological effects, such as central nervous system (CNS) effects, cardiovascular effects, renal effects, respiratory effects, immunological effects, gastrointestinal effects, and metabolic effects. The relative amount of adenosine in an individual may be associated with the effects listed below, such that elevated levels of adenosine may stimulate a particular effect, such as an unwanted physiological response, such as an asthma attack.

TsNeO effects are a decrease in the level of transmitter release (A 4), a sedative effect (A5), reduced motor activity (Aza), anticonvulsant activity, stimulation of chemoreceptors (A 5) and hyperalgesia. Therapeutic applications of the compounds of this invention are the treatment of dementia and Alzheimer's disease and the improvement of memory. 70 Cardiovascular effects are vasodilation (Aza), (Azu) and (Az), vasoconstriction (A 4), bradycardia (A), inhibition of platelets (A 24), negative inotropy and dromotropic heart (A 4), arrhythmia , tachycardia and angiogenesis.

Therapeutic applications of the compounds of this invention are, for example, prevention of ischemia-induced damage to the heart and reduction of its tone, protection of myocardial tissue and restoration of cardiac function.

Renal-like effects are a decrease in GFR (glomerular filtration rate) (A 4), shrinkage of /5 mesangial cells (A;), antidiuresis (A) and inhibition of renin release (A 4). Suitable therapeutic uses of the compounds of this invention are their use as diuretics, natriuretics, potassium-sparing agents, agents for protecting/preventing kidneys from acute renal failure, hypotensive, anti-edematous and anti-nephritic agents.

Respiratory effects are broncholytic effects (A"), bronchostenosis (A)), chronic obstructive pulmonary disease, allergic rhinitis, mucus secretion and respiratory depression (A 5). Suitable therapeutic applications for the compounds of this invention are their use as anti-asthmatic agents, treatment of post-transplant lung diseases and respiratory diseases.

The immunological effects are immunosuppression (A5), chemotaxis of neutrophils (A5), generation of superoxide by neutrophils (Aza) and degranulation of mast cells (A»2j and Az). The therapeutic use of antagonists is their common use as mediators of allergic and non-allergic inflammatory reactions, for example, mediators of the release of histamine and other mediators of inflammation.

Gastrointestinal effects are inhibition of acid secretion (A). The therapeutic application of the compounds of this invention is their use in reflux conditions and ulcers. Gastrointestinal effects also include diseases of the large intestine, small intestine and diarrhea, for example, diarrhea associated with inflammation of the small intestine Ge (Az).

Eye diseases include damage to the retina and optic disc, and diseases associated with injuries and, (Az). In a preferred embodiment of the invention, such eye disease is glaucoma. M

Other therapeutic applications of the compounds of this invention are the treatment of obesity (due to the lipolytic properties of these compounds), hypertension and depression; use as a sedative, tranquilizer, p

As an antileptic agent and as a laxative, for example, to stimulate peristalsis, which does not lead to diarrhea. uh-

The term "disease state" means conditions caused by or associated with undesirable levels of adenosine, adenylyl cyclase activity, increased physiological activity associated with impaired stimulation of adenosine receptors and/or with the growth of CAMP. In one embodiment of the invention, the pathological condition is, for example, asthma, chronic obstructive pulmonary disease, allergic rhinitis, bronchitis, kidney disease, gastrointestinal disease or eye disease. Other examples are chronic bronchitis and bladder fibrosis. Relevant examples of inflammatory diseases are non-lymphatic leukemia, myocardial ischemia, angina pectoris, heart attack, cerebrovascular ischemia, intermittent claudication, threatening ischemia. limbs, venous hypertension, varicose veins, venous occlusion and arteriosclerosis. Injuries associated with reperfusion include, for example, any injury after surgery, such as reconstructive

Surgical operation, thrombolysis or angioplasty. -I The term "effect on a condition responsive to M-6b-substituted 7-deazapurine" or "treatment of a condition responsive to M-b-substituted 7-deazapurine" means such changes in the pathological condition or disease described above, which may lead to reduction or minimization of physiological symptoms in the mammal. This -I expression also means regulation, prevention or inhibition of physiological symptoms or effects associated with an abnormal amount of adenosine. In a preferred embodiment of this invention, such regulation of a pathological condition or disease is its complete treatment. In another preferred embodiment of this invention,

Ф this regulation is chosen in such a way that control over the deviation of adenosine receptor activity levels is carried out, but, at the same time, other physiological systems and parameters remain unchanged.

The term "M-6-substituted 7-deazapurine" is understood by specialists and means compounds having the formula I: 29 Creams. o M-6 In yav "7 deata vtse" and br in Iya 60

In,

FO

"M-substituted 7-deazapurine" includes pharmaceutically acceptable salts thereof, and, in one embodiment, also refers to some of the M-6-substituted purines described herein. 65 In some embodiments of the invention, M-6-substituted 7-deazapurine is not substituted with M-b-benzyl or

M-6E-phenylethyl. In other variants of the invention, K. is not substituted by benzyl or phenylethyl. In preferred embodiments of the invention, K. and K" are both non-hydrogen atoms. In other preferred embodiments, Kz is not a hydrogen atom.

The term "therapeutically effective amount" of an M-6-substituted 7-deazapurine, as described below, means an amount of the therapeutic compound necessary or sufficient to achieve its intended function in a mammal, e.g., to treat a condition responsive to M-6-substituted 7 -deazapurine, or a pathological condition in a mammal. The effective amount of a therapeutic compound may vary depending on factors such as the amount of etiologic agent already present in the mammal, the age, sex, and weight of the mammal, and the ability of the therapeutic compounds of the present invention to induce a condition responsive to M-6-substituted deazapurine in a mammal Each person skilled in the art can independently investigate the above factors and determine the effective amount of 7/0 therapeutic compound without any undue experimentation. IP migo- and IP mimo-assays can be used to determine the "effective amount" of the therapeutic compounds described below. Each person skilled in the art can select an appropriate amount of a therapeutic compound for use in the above assay or for therapeutic treatment.

A therapeutically effective amount preferably attenuates at least one symptom or effect associated with the u/5 btan or disease responsive to the action of M-6b-substituted 7-deazapurine, and which is amenable to treatment by at least about 2095 (more preferably, at least , by about 4095, even more preferably by at least about 6055, and most preferably by at least about 8095) compared to the symptom or effect in an individual not amenable to such treatment. Assays to determine the degree of attenuation of such symptoms and/or effects can be developed by any expert. Any known assay capable of determining such 20 parameters is part of the present invention. Thus, for example, if the disease to be treated is asthma, the volume of air exhaled from the individual's lungs can be measured before and after treatment to determine the increase in this volume by a method known to those skilled in the art. Similarly, if the disease to be treated is an inflammatory disease, then to determine the reduction in the area of inflammation, this area of inflammation can be measured before and after treatment by a method known to those skilled in the art. high school

The term "cell" refers to both a prokaryotic cell and a eukaryotic cell.

The term "animal" means any organism with adenosine receptors or any organism i) susceptible to a condition that responds to M-6-substituted 7-deazapurine. Examples of such animals are yeast, mammals, reptiles and birds. This concept also includes transgenic animals.

The term "mammal" is understood by a person skilled in the art and means an animal, preferably a warm-blooded animal, and most preferably cattle, sheep, pigs, horses, dogs, cats, rats, mice and humans. Also included within the scope of this invention are, for example, mammals susceptible to administration-responsive states and,

M-6-substituted 7/-deazapurine, inflammatory diseases, emphysema, asthma, diseases of the central nervous system M or acute respiratory distress syndrome.

In another aspect, the present invention relates to methods of modulating adenosine receptor(s) in mammals

335 By administering to said mammal a therapeutically effective amount of M-b-substituted 7-deazapurine, so that this administration leads to modulation of the adenosine receptor in the mammal. Suitable adenosine receptors are receptors of families A, A" or Az. In a preferred embodiment, M-b-substituted 7-deazapurine is an adenosine receptor antagonist.

The term "modulation of an adenosine receptor" refers to events in which a compound interacts with an adenosine receptor(s), causing increased, decreased, or abnormal physiological activity associated with the adenosine receptor, or a subsequent cascade of reactions induced as a result modulation. adenosine receptor. Physiological activity associated with the adenosine receptor is the induction of a sedative effect, dilation of blood vessels, reduction of heart rate and contractility of the heart, inhibition of platelet aggregation, stimulation of gluconeogenesis, inhibition of lipolysis, opening of potassium channels, reduction of the flow of calcium ions in calcium channels, etc. p. -I The terms "modulate", "modulation" or "modulation" mean the prevention, elimination or inhibition of a cumulative increase in unwanted physiological activity associated with abnormal stimulation of the adenosine receptor, for example, using the therapeutic methods of this invention. In another -I variant of this invention, the term "modulation" means antagonistic effects, for example, reducing the activity or production of mediators of allergy or allergic inflammation, caused by overstimulation of adenosine receptor(s). Thus, for example, therapeutic deazapurines of this invention can

Ф interact with the adenosine receptor with inhibition, for example, of adenylate cyclase activity.

The term "condition characterized by abnormal activity of the adenosine receptor" means a disease, disorder or condition associated with impaired stimulation of the adenosine receptor, such that stimulation of the receptor causes a cascade of biochemical or physiological reactions that are directly or indirectly associated with the disease. disorder or condition. This stimulation of the adenosine (F) receptor is not the sole etiological factor of a given disease, disorder or condition, but it is only responsible for the occurrence of some of the symptoms usually associated with a given disease, disorder or treatable condition. Such disruption of receptor stimulation may be the only factor, or at least one other agent, that may cause the condition requiring treatment. Examples of such conditions are the conditions listed above, including inflammation, gastrointestinal disease, and symptoms manifested in the presence of increased adenosine receptor activity. Preferred examples are symptoms associated with asthma, allergic rhinitis, chronic obstructive pulmonary disease, emphysema, bronchitis, gastrointestinal disease, and glaucoma. 65 The term "therapy or treatment of a condition characterized by impaired adenosine receptor activity" means the alleviation or attenuation of at least one symptom associated with in this state.

Such treatment is also the alleviation or attenuation of more than one symptom. Preferably, if such treatment leads to a cure, for example mainly to the elimination of symptoms associated with the given condition.

The present invention relates to compounds, M-6-substituted 7-deazapurines, having the formula I:

When in

M x b re | In, IN

K,

F where: each of K. and Co independently represents a hydrogen atom or a substituted or unsubstituted alkyl, aryl or 72 alkylaryl group, or, taken together, they form a substituted or unsubstituted heterocyclic ring;

Ez represents a hydrogen atom or a substituted or unsubstituted alkyl, aryl or alkylaryl group;

KE. represents a hydrogen atom or a substituted or unsubstituted alkyl, aryl or alkylaryl group. Each of K5 and Co independently represents a halogen atom, for example, chlorine, fluorine or bromine, a hydrogen atom or a substituted or unsubstituted alkyl, aryl or alkylaryl group; or, K and K5 or K»b and Co, taken together, will form a substituted or unsubstituted heterocyclic or carbocyclic ring. The present invention also relates to pharmaceutically acceptable salts of M-6-substituted 7-deazapurines.

In some embodiments of the invention, each of K 4 and Co can independently represent substituted or unsubstituted cycloalkyl or heteroarylalkyl groups. In other embodiments of the invention, K c means a hydrogen atom or a substituted or unsubstituted heteroaryl group. In other embodiments of the invention, each SM

Wu, WB and Be can independently represent a heteroaryl group. d)

In one embodiment of the invention, K is a hydrogen atom, K is a substituted or unsubstituted cyclohexane, cyclopentyl, cyclobutyl, or cyclopropane group, Kz is a substituted or unsubstituted phenyl group, K is a hydrogen atom, and Kb and K5 are both are methyl groups. X,

In another embodiment of the invention, K is cyclohexanol, cyclohexanediol, cyclohexylsulfonamide, cyclohexanamide, cyclohexyl ester, cyclohexene, cyclopentanol or cyclopentanediol, and Kz is a phenyl group. uh-

In another embodiment of the invention, K is a hydrogen atom, Co is cyclohexanol, K 3 is a substituted or unsubstituted phenyl, pyridine, furan, cyclopentane or thiophene group, B; represents a hydrogen atom, a substituted alkyl, aryl or arylalkyl group, and each of B in Kv i- independently represents a hydrogen atom or a substituted or unsubstituted alkyl, aryl or alkylaryl group.

In another embodiment of the invention, K 4 represents a hydrogen atom, K o represents a substituted or unsubstituted alkylamine, arylamine or alkylarylamine, substituted or unsubstituted alkylamide, arylamide or alkylarylamide, substituted or unsubstituted alkylsulfonamide, arylsulfonamide or alkylarylsulfonamide, substituted or unsubstituted alkylurea, arylurea or alkylarylurea, substituted or unsubstituted C with alkylcarbamate, arylcarbamate or alkylarylcarbamate, or substituted or unsubstituted alkylcarboxylic acid, "» arylcarboxylic acid or alkylarylcarboxylic acid, Kz represents a substituted or unsubstituted phenyl " group, K. represents a hydrogen atom, and Kb and K5 are throwing groups.

In another embodiment of the invention, Ke is guanidine, modified guanidine, cyanoguanidine, thiourea, thioamide, or amidine. - In one of the variants of implementation of the invention, Ko can be: o V. - t M a; with Ki Koi 8) where each of Koa-Kos independently represents a hydrogen atom or a saturated or unsaturated alkyl, aryl or alkylaryl group, and Code represents a hydrogen atom or a saturated or unsaturated aalkyl, aryl or alkylaryl group, MKoeKoi or OK", where each of Coe-K29 independently represents a hydrogen or saturated atom

F) or an unsaturated alkyl, aryl or alkylaryl group. Alternatively, Coa and Coa taken together can form a carbocyclic or heterocyclic ring, one having about 3-8 members, eg, cyclopropyl, cyclopentyl, cyclohexyl groups. In one aspect of the present invention, Kb and Ke are both non-methyl groups, and preferably one of Kb and Ke is an alkyl group, such as a methyl group, and the other is a hydrogen atom.

In another aspect of the present invention, if the CC. is 1-phenylethyl and K 4 is hydrogen, then K 2 is not phenyl, 2-chlorophenyl, 3-chlorophenyl, 4-chlorophenyl, 3,4-dichlorophenyl, 3-methoxyphenyl or 4-methoxyphenyl, or if K and K. represent 1-phenylethyl, then Kz is not a hydrogen atom, or if K; 65 represents a hydrogen atom, and Kz represents phenyl, then K. is not phenylethyl.

In another aspect of this invention, if K»5 and Kv taken together form a carbocyclic ring, for example:

Kg 7 x

And | M

IN

KE, then pyrimido|4,5-b|indole, then Kz is not phenyl, and if K is 1-(4-methylphenyl)ethyl, phenylisopropyl, phenyl or 1-phenylethyl or if Kz is not a hydrogen atom, then K. is 1-phenylethyl. The carbocyclic ring formed by K5 and Kv can be either aromatic or aliphatic and can have from 4 to 12 carbon atoms, for example, naphthyl, phenylcyclohexyl, etc., and preferably from 5 to 7 carbon atoms, for example, cyclopentyl or cyclohexyl. Alternatively, K; and Co taken together can form a heterocyclic ring such as the t ring described below. Common heterocyclic rings are rings having from 4 to 12 carbon atoms, preferably from 5 to 7 carbon atoms, and these rings can be either aromatic or aliphatic.

This heterocyclic ring can be, in addition, substituted, including the replacement of one or more carbon atoms of the ring structure by one or more heteroatoms.

In yet another aspect of this invention, K.; and Co will form a heterocyclic ring. Typical examples include, but are not limited to, the heterocyclic rings listed below, such as morpholino, piperazine, and the like, such as 4-hydroxypiperidines, 4-aminopiperidines. If K. and Co, taken together, form a piperazine group, then KU can be a hydrogen atom or a substituted or unsubstituted alkyl, aryl or alkylaryl

M. o sia

About the group.

In yet another aspect of the present invention, CC; and Co taken together can form a heterocyclic ring, for example: Ge)

Co

KE, shcha ih and re Lo cm z6 M 0 i-

The heterocyclic ring may be either aromatic or aliphatic and may form a ring having from 4 to 12 carbon atoms, e.g., naphthyl, phenylcyclohexyl, etc., and may be either aromatic or aliphatic, e.g., cyclohexyl, cyclopentyl . The heterocyclic ring can be, in addition, substituted, including the replacement of the carbon atoms of the ring structure by one or more heteroatoms. Alternatively, c PE and K5 taken together can form a heterocyclic ring, such as the ring described below. In some embodiments of the invention, the M-b-substituted 7-deazapurine is not M-b-benzyl- or "» M-6-phenylethyl-substituted. In other embodiments of the present invention, K. is not benzyl- or phenylethyl-substituted. In preferred embodiments embodiment of the invention, K. and Co are both non-hydrogen atoms. In another preferred 5 embodiment of the invention, Kz is not a hydrogen atom. -I Compounds of the present invention may contain water-soluble prodrugs which, as a result of metabolism, are converted into an active medicinal substance, e.g. , as a result of esterase-catalyzed hydrolysis. d Examples of potential prodrugs are deazapurines, where, for example, Co is cycloalkyl substituted -OC(O) -I (2MH", where 7 is the side chain of a natural or unnatural amino acid or its analog . carbon acid, p-alanine, y-aminobutyric acid, alanine-alanine or glycine-alanine.

In another preferred embodiment, the present invention relates to diazapurines of formula (I), where:

Cu is hydrogen;

Co is substituted or unsubstituted cycloalkyl, substituted or unsubstituted alkyl, or K 4 and K»

GF! taken together, form a substituted or unsubstituted heterocyclic ring;

Ez is unsubstituted or substituted aryl; ko KE. represents hydrogen and each of Kb and Co independently represents hydrogen or alkyl; as well as their pharmaceutically acceptable salts. In this version of the present invention, these deazapurines can be selective antagonists of the Az receptor.

In one of the variants of this invention, Co is a substituted (for example, hydroxy-substituted) or unsubstituted cycloalkyl. In a preferred embodiment, K and K are hydrogen, K is unsubstituted or substituted phenyl, and each of Kb and Ke is alkyl. Preferably, Co is mono-hydroxycyclopentyl or mono-hydroxycyclohexyl. Co may also be substituted with -MH-C(-O)E, where E is substituted or unsubstituted C-C-alkyl (eg, alkylamine, eg, ethylamine).

Ku and Co taken together can form a substituted or unsubstituted heterocyclic ring, which can be substituted with an amine or an acetamido group.

In another aspect of the invention, Co can be -A-MHNOS(-O)B, where A is unsubstituted C 4 -C 4 alkyl (eg, ethyl, propyl, butyl) and B is substituted or unsubstituted C 4 -C 4 alkyl ( for example, methyl, aminoalkyl, for example, aminomethyl or aminoethyl, alkylamino, for example, methylamino, ethylamino), and preferably, if K. and K. represent hydrogen, then K»z represents a substituted or unsubstituted phenyl, and each of K5 and Ke is alkyl. "B" can be substituted or unsubstituted cycloalkyl (eg, cyclopropyl or 1-aminocyclopropyl). 70 In another variant of this invention, Kz can be a substituted or unsubstituted phenyl, and preferably, each of K» and Ko is an alkyl. Preferably, K c can have one or more substituents (for example, o-, m- or p-chlorophenyl, o-, m- or p-fluorophenyl).

Ez can preferably be a substituted or unsubstituted heteroaryl, and preferably each of Kb and Kb is alkyl. Examples of heteroaryl groups are pyridyl, pyrimidyl, pyridazinyl, pyrazinyl, pyrrolyl, triazolyl, thiazolyl, oxazolyl, oxadiazolyl, furanyl, methylenedioxyphenyl and thiophenyl. KZ is preferably 2-pyridyl, 3-pyridyl, 4-pyridyl, 2-pyrimidyl or 3-pyrimidyl.

In one of the preferred embodiments of the invention, each of K 5 and Co is hydrogen. In another embodiment, each of K" and K5 is methyl.

In a particularly preferred variant, the deazapurines of this invention are water-soluble prodrugs that, as a result of metabolism, are converted into an active medicinal substance, for example, as a result of esterase-catalyzed hydrolysis. Preferred prodrugs contain the group K 5", which is a cycloalkyl substituted -OC(O) (2МН", where 7 is the side chain of a natural or unnatural amino acid or its analogue, o-, p-, y- or 5o-amino acid or The preferred aminooxylate side chains are those of glycine, alanine, valine, leucine, isoleucine, lysine, o-methylalanine, aminocyclopropanecarboxylic acid, su azetidine-2-carboxylic acid, p-alanine, γ-aminobutyric acid, alanine-alanine, or glycine -alanine. o

In a particularly preferred embodiment, 7 is a side chain of glycine, K» is cyclohexyl, K c is phenyl, and K5 and Ko are methyl.

In a preferred embodiment of the present invention, the deazapurine is 4-(cis-3-hydroxycyclopentyl)-amino-5,6-dimethyl-2-phenyl-7H-pyrrolo|2,Za|pyrimidine. (Se)

In another embodiment of the present invention, deazapurine is a salt of trifluoroacetic acid and c 4-(cis-3-(2-aminoacetoxy)cyclopentyl)amino-5,6-dimethyl-2-phenyl-7H-pyrrolo|2,3a|pyrimidine.

In another embodiment of the present invention, deazapurine is 1-4-(3-acetamido)piperidinyl,-5,6-dimethyl-2-phenyl-7H-pyrrolo-(2,34|pyrimidine.

In another embodiment of the present invention, deazapurine is d-(2-M'-methylcarbomidopropyl)amino-5,6-dimethyl-2-phenyl-7H-pyrrolo(|2,Za|pyrimidine. -

In another embodiment of the present invention, deazapurine is 4-(2-acetamidobutyl)amino-5,6-dimethyl-2-phenyl-7H-pyrrolo|2,3a|-pyrimidine.

In another embodiment of the present invention, the deazapurine is d- (2-M'-methylcarbamidobutyl)amino-5,6-dimethyl-2-phenyl-7H-pyrrolo|2,Za|pyrimidine.

In another embodiment of the present invention, the deazapurine is 4-(2-aminocyclopropylacetamidoethyl)amino-2-phenyl-7H-pyrrolo-(2,Za|pyrimidine). In another embodiment of the present invention, the deazapurine is 4-( trans-4-hydroxycyclohexyl)amino-2-(3-chlorophenyl)-7H-pyrrolo(2534d|pyrimidine.

In another embodiment of the present invention, deazapurine is 4-(trans-4-hydroxycyclohexyl)amino-2-(3-fluorophenyl)-7H-pyrrolo|2,Z|pyrimidine. it. In another embodiment of the present invention, deazapurine is 4-(trans-4-hydroxycyclohexyl)amino-2-(4-pyridyl)-7H-pyrrolo (2,34|pyrimidine.

In another variant of its implementation, the present invention relates to a method of inhibiting the activity of an adenosine receptor (for example, Az, Aod, Aov or preferably, Az) in a cell by contacting this cell with M-6-substituted 7-deazapurine (for example, preferably , with an adenosine receptor antagonist).

In another aspect of its implementation, the present invention relates to a method of treating eye damage in an animal (eg, a human) by administering to the animal an effective amount of M-6b-substituted 7-deazapurine.

The preferred M-b-substituted 7-deazapurine is an antagonist of Az adenosine receptors in animal cells.

The specified eye lesion is a retinal or optic disc lesion, and this lesion may be acute or chronic. This lesion can occur as a result of glaucoma, edema, ischemia, hypoxia, or trauma.

GF) In a preferred embodiment, the present invention relates to a deazapurine of formula II, see above, where: o X represents M or SC; each of C and C is independently hydrogen or substituted or unsubstituted alkoxy, aminoalkyl, alkyl, aryl or alkylaryl, or taken together they form a substituted or unsubstituted heterocyclic ring, provided that both C and C are not hydrogen;

Ez is substituted or unsubstituted alkyl, arylalkyl or aryl;

KE. represents hydrogen or substituted or unsubstituted S.-Svalkil;

Y represents hydrogen, substituted or unsubstituted alkyl, or K; and I, taken together, form a substituted or unsubstituted heterocyclic or carbocyclic ring;

Ke is hydrogen, substituted or unsubstituted alkyl or halogen;

O is CH", O, Z or MK), where K; represents hydrogen or substituted or unsubstituted S.-Svalkil and

M/ represents unsubstituted or substituted alkyl, cycloalkyl, alkynyl, aryl, arylalkyl, biaryl, heteroaryl, substituted carbonyl, substituted thiocarbonyl or substituted sulfonyl; provided that if Kz is pyrrolidino, then K. is not methyl.

In one of the variants of this invention, in the compounds of formula II, X represents SK and O represents CH", 0, Z or MH. In another variant of the invention, X is M.

In another version of the compounds of formula II, MU is a substituted or unsubstituted aryl, 5- or β-yl 7/0 heteroaryl or biaryl. M/ can be replaced by one or several deputies. Examples of substituents are: halogen, hydroxy, alkoxy, amino, aminoalkyl, aminocarboxyamide, SM, SEz, СО»Ка, СОМНЕК»в, СОМ», ZOKv,

Ov, and ZO2MKaCo»o, where each of Ka and KO independently represents hydrogen or substituted or unsubstituted alkyl, cycloalkyl, aryl or arylalkyl. Preferably, M/ can be substituted or unsubstituted phenyl, for example, methylenedioxyphenyl. M/ can also represent a substituted or unsubstituted 5-membered heteroaryl ring, for example, pyrrole, pyrazole, oxazole, imidazole, triazole, tetrazole, furan, thiophene, thiazole, and oxadiazole.

Preferably, MU can be a 6-membered heteroaryl ring, for example, pyridyl, pyrimidyl, pyridazinyl, pyrazinyl and thiophenyl. In a preferred embodiment of the invention, MU is 2-pyridyl, 3-pyridyl, 4-pyridyl, 2-pyrimidyl, 4-pyrimidyl or 5-pyrimidyl.

In one preferred embodiment of the compounds of formula II, CO) represents MH, and UU represents a 3-pyrazole ring, which is unsubstituted or M-substituted by substituted or unsubstituted alkyl, cycloalkyl, aryl or arylalkyl.

In another preferred embodiment of the compounds of formula II, O is oxygen and MU is a 2-thiazole ring that is unsubstituted or substituted with substituted or unsubstituted alkyl, cycloalkyl, aryl or arylalkyl. high school

In another variant of the compounds of formula II, M/) represents substituted or unsubstituted alkyl, cycloalkyl, for example, cyclopentyl or arylalkyl. Examples of substituents are halogen, hydroxy, substituted or unsubstituted i) alkyl, cycloalkyl, aryl arylalkyl or MNK.0, where Co is hydrogen, or substituted or unsubstituted alkyl, cycloalkyl, aryl or arylalkyl.

In another variant of its implementation, the present invention relates to a deazapurine of the formula ІІ, where MU is Hezo 7 "СН Э(О) У or - (СНо)а-С(-5) М, where a is equal to a whole number from 0 to C, M represents aryl, alkyl, arylalkyl, cycloalkyl, heteroaryl, alkynyl, МНА..4К.", or if O represents MH, then U represents o

OK.z, and where each of K.44, K." and C is independently hydrogen or unsubstituted or substituted alkyl, aryl, n-arylalkyl or cycloalkyl. U is preferably a 5- or b-membered heteroaryl ring.

In addition, MU is -(CH 2)5-C(-О0)/U where ) is equal to 1 or 2, B is equal to 0, 1, 2 or 3, and U is aryl, alkyl, arylalkyl, cycloalkyl, alkynyl, heteroaryl, MNK.4K.5 or in the case that B is equal to 1, 0) is CH", and where each of Ka, K.5 and K416b independently represent hydrogen or unsubstituted or substituted alkyl, aryl, arylalkyl or cycloalkyl.

In another embodiment of the invention, Kz is selected from the group consisting of substituted and unsubstituted phenyl, pyridyl, pyrimidyl, pyridazinyl, pyrazinyl, pyrrolyl, triazolyl, thiazolyl, oxazolyl, oxadiazolyl, pyrazolyl, furanyl, methylenedioxyphenyl and thiophenyl. z c If Kz is phenyl, it may be substituted, for example, with hydroxyl, alkoxy (eg, methoxy), alkyl (eg, tolyl), and halogen (eg, o-, m-, or p-fluorophenyl or o-, m- or y?" p-chlorophenyl). Kz preferably can be 2-, 3- or 4-pyridyl or 2- or 3-pyrimidyl.

The present invention also relates to deazapurine, where Co is hydrogen or C.4-Szalkyl. Ke is mostly hydrogen. -I The present invention also relates to deazapurines, where K is hydrogen and Co is substituted or unsubstituted alkyl or alkoxy, substituted or unsubstituted alkylamine, arylamine or alkylarylamine, substituted or unsubstituted aminoalkyl, aminoaryl or aminoalkylaryl, substituted or unsubstituted alkylamide, arylamide -I or alkylarylamide, substituted or unsubstituted alkylsulfonamide, arylsulfonamide or 5-alkylarylsulfonamide, substituted or unsubstituted alkylurea, arylurea or alkylarylurea, o substituted or unsubstituted alkylcarbamate, arylcarbamate or alkylarylcarbamate, or substituted or

F unsubstituted alkylcarboxylic acid, arylcarboxylic acid or alkylarylcarboxylic acid. Co is preferably a substituted or unsubstituted cycloalkyl, for example, mono- or dihydroxy-substituted cyclohexyl or cyclopentyl (preferably, monohydroxy-substituted cyclohexyl or mono-hydroxy-substituted cyclopentyl).

Preferably, Ko can have the formula: о Хи ю АСК ZVO -А b ОВ

Kk aso 60 where A is C 4-Salkyl, C3--SUcycloalkyl, a chain of 1-7 atoms or a ring of 3-7 atoms, optionally substituted with C-Salkyl, halogen, hydroxyl, carboxyl, thiol or amino groups;

B is methyl, M(Me) », MEO», MNMe, MNE, (CHO) MNac, MH(CHO) CH», (CH) MH», (CH) СНеН»МН», (сноХМНМе, (СНОХОН, СНСМ, (СНо)тсОоНн, СНКв8Кло or СНМЕон, where r is an integer from 0 to 2, t 65 is equal to 1 or 2, Kw is alkyl, Kw is MH or СОН, or Kw and Kw taken together are teka (SNI) where p equals 2 or Z i

Ki; is C 41-Salkyl, C3-Ccycloalkyl, a chain of 1-7 atoms or a ring of 3-7 atoms, optionally substituted with C-Salkyl, halogen, hydroxyl, carboxyl, thiol or amino groups. "A" is mostly unsubstituted or substituted S.-Svalkil. "B" can be unsubstituted or substituted S.-Svalkil. 70 In a preferred embodiment of the invention, K » represents the formula -А-МНОС(-О)В. In a particularly preferred variant of the invention, A is -CHNoCH»-, and B is methyl.

The compounds of the present invention may contain water-soluble prodrugs, which, as a result of metabolism, are converted into an active medicinal substance, for example, as a result of esterase-catalyzed hydrolysis.

Examples of potential prodrugs are deazapurines, where, for example, Co is a cycloalkyl substituted with 75 7ОС(ОХ2)МНо, where and is the side chain of a natural or unnatural amino acid or its analogue, o-, D-, y- or 5-amino acid or dipeptide. Preferred aminooxylate side chains are glycine, alanine, valine, leucine, isoleucine, lysine, o-methylalanine, aminocyclopropanecarboxylic acid, azetidine-2-carboxylic acid, p-alanine, y-aminobutyric acid, alanine-alanine, or glycine-alanine.

In another embodiment of the invention, K. and Co., taken together, represent: h where n is equal to 1 or 2 and where the specified ring is possibly, but not necessarily, substituted by one or more hydroxyl, amino, thiol, carboxyl, halogen, СНоОНН, СНоМНОо(-О)-alkyl or "С

СНоМНОТ(О)МН with alkyl groups. (5)

In one of the preferred embodiments of the invention, Ki is hydrogen, Ko is a substituted or unsubstituted S.-Svalkyl, Kz is a substituted or unsubstituted phenyl, K is hydrogen, !. represents hydrogen or substituted or unsubstituted C.-Svalkyl, O) represents O, 5 or MK 7, where K represents hydrogen or substituted or unsubstituted C.-Svalkyl, and M/ represents substituted or unsubstituted aryl. (Se)

Ро is preferably -А-МНО(-О)В, where each of A and В is independently unsubstituted or substituted with

C.-C,-alkyl. So, for example, A can be SNOSN. B can be, for example, alkyl (eg, methyl) or aminoalkyl (eg, aminomethyl). Kz is preferably unsubstituted phenyl, and Y is hydrogen. Co can be methyl or, preferably, hydrogen. O is preferably O, 5 or MK 7, where K7 is hydrogen or substituted or unsubstituted C 4-alkyl, for example, methyl. MU is unsubstituted or substituted phenyl (for example, alkoxy-, halogen-substituted). Preferably, MU is - p-fluorophenyl, p-chlorophenyl or p-methoxyphenyl. MU can also be heteroaryl, for example, 2-pyridyl.

In a particularly preferred embodiment of the invention, the indicated deazapurine is 4-(2-acetylaminoethyl)-amino-β6-phenoxymethyl-2-phenyl-7H-pyrrolo-[2,3]pyrimidine. "

In a particularly preferred embodiment of the invention, the indicated deazapurine is 4-(-2-acetylaminoethyl)-amino-6--4-fluorophenoxy)methyl-2-phenyl-7H-pyrrolo(|2,Za|pyrimidine. - with especially in a preferred embodiment of the invention, the specified deazapurine is also 4-(2-acetylaminoethyl)-amino-6-(4-chlorophenoxy)methyl-2-phenyl-7H-pyrrolo|2,3a|pyrimidine." In a particularly preferred embodiment of the invention , the indicated deazapurine is 4-(2-acetylaminoethyl)-amino-6-(4-methoxyphenoxy)methyl-2-phenyl-7H-pyrrolo-(2,Za|pyrimidine).

In a particularly preferred embodiment of the invention, the indicated deazapurine is 4-(2-acetylaminoethyl)-amino-6-(2-pyridyloxy)methyl-2-phenyl-7H-pyrrolo|2,Z|pyrimidine. with In a particularly preferred embodiment of the invention, the specified deazapurine is 4-(2-acetylaminoethyl)-amino-6-(M-phenylamino)methyl-2-phenyl-7H-pyrrolo-(2,34|pyrimidine. -and in particular in a preferred embodiment of the invention, the specified deazapurine is 4--2-acetylaminoethyl)-amino-6-((M-methyl-M-phenylamino)methyl-2-phenyl-7H-pyrrolo|2,Za|pyrimidine.

In a particularly preferred embodiment of the invention, the indicated deazapurine is 4) 4-(2-M'-methylcarbamidoethyl)amino-6-phenoxymethyl-2-phenyl-7H-pyrrolo|2,Za|pyrimidine.

The present invention also relates to a method of inhibiting the activity of an adenosine receptor (for example, an adenosine receptor Az) in cells by contacting said cell with a compound of the present invention.

A preferred compound is a receptor antagonist. o The present invention also relates to a method of treating a gastrointestinal disease (e.g., diarrhea) in an animal by administering to the animal an effective amount of a compound of the present invention (e.g., an Az antagonist). The dominant animal is man.

In another variant of its implementation, the present invention relates to a composition containing the M-6-substituted 7-deazapurine of the present invention and a pharmaceutically acceptable carrier.

The present invention relates to a method of treating an M-6-substituted 7-deazapurine responsive condition in an animal by administering to a mammal a therapeutically effective amount of a deazapurine of the present invention necessary to treat an M-6-substituted 7-deazapurine responsive condition in an animal . The specified pathological condition can preferably be a disease mediated by adenosine. Predominant examples of pathological b5 conditions are: diseases of the central nervous system, cardiovascular diseases, renal diseases, inflammatory diseases, allergic diseases, gastrointestinal diseases, eye diseases and respiratory diseases.

The term "alkyl" means a radical of saturated aliphatic groups, including straight-chain alkyl groups, branched-chain alkyl groups, cycloalkyl (alicyclic) groups, alkyl-substituted cycloalkyl groups and cycloalkyl-substituted alkyl groups. The term "alkyl" further means alkyl groups which may additionally include oxygen, nitrogen, sulfur or phosphorus atoms replacing one or more carbon atoms of the hydrocarbon backbone, for example, oxygen, nitrogen, sulfur or phosphorus atoms. In preferred embodiments of the invention, the straight or branched chain alkyl has 30 or fewer carbon atoms on its backbone (eg, C4-C20 for a straight chain, C3-C20 for a branched chain) and more preferably, 20 carbon atoms or less . Similarly, preferred cycloalkyls have from 4 to 10 carbon atoms in their ring structure, and more preferably, 5, 6 or 7 carbon atoms in their ring structure.

Moreover, the term "alkyl" used in the description and in the claims includes both "unsubstituted alkyls" and "substituted alkyls", where the indicated substituted alkyls mean alkyl groups having substituents replacing hydrogen on one or more atoms carbon of the hydrocarbon backbone. Such substituents /5 can be, for example, halogen, hydroxyl, alkylcarbonyloxy-, arylcarbonyloxy-, alkoxycarbonyloxy-, aryloxycarbonyloxy-, carboxylate, alkylcarbonyl, alkoxycarbonyl, aminocarbonyl, alkylthiocarbonyl, alkyloxy, phosphate, phosphonato-, phosphinato-, cyano-, amino- (include alkylamino-, dialkylamino-, arylamino-, diarylamino- and alkylarylamino-) acylamino- (including alkylcarbonylamino-, arylcarbonylamino-, carbamoyl and ureido-), amidino-, imino-, sulfhydryl, alkylthio-, arylthio-, thiocarboxylate, sulfate , sulfonato-, sulfamoyl, sulfonamido-, nitro-, trifluoromethyl, cyano-, azido-, heterocyclyl, alkylaryl, or aromatic or heteroaromatic groups. It should be noted that these substituted groups on the hydrocarbon chain can themselves be substituted if necessary. Cycloalkyls can also be substituted, for example, by the substituents described above. The term "alkylaryl" group means alkyl substituted with aryl (eg, phenylmethyl (benzyl)). The term "alkyl" also means unsaturated aliphatic groups having a length and possible substituents similar to the alkyls described above, but which contain at least one double or triple bond, respectively. (8)

The term "aryl" as used herein means the radical of aryl groups, including 5- and b--substituted aromatic groups with one ring, which may have from 0 to 4 heteroatoms, for example, benzene, pyrrole, furan, thiophene, imidazole, benzoxazole, benzothiazole, triazole, tetrazole, pyrazole, pyridine, pyrazine, pyridazine and pyrimidine and Ge

Zr T.P. Aryl groups are also polycyclic condensed aromatic groups, such as naphthyl, quinolyl, indolyl, etc. These aryl groups having heteroatoms on their ring structure may also be referred to as "aryl heterocycles", "heteroaryls" or "heteroaromatic groups". This aromatic ring may M be substituted in one or more positions of the ring by the substituents described above, for example, such as halogen, hydroxyl, alkoxy, alkylcarbonyloxy-, arylcarbonyloxy-, alkyloxycarbonyloxy-, aryloxycarbonyloxy-, carboxylate, alkylcarbonyl, alkoxycarbonyl, aminocarbonyl, and - alkylthiocarbonyl, phosphate, phosphonato-, phosphinato-, cyano-, amino- (including alkylamino-, dialkylamino-, arylamino-, diarylamino- and alkylarylamino-), acylamino-(including alkylcarbonylamino-, arylcarbonylamino-, carbamoyl and ureido-), amidino-, imino-, sulfhydryl, alkylthio-, arylthio-, thiocarboxylate, sulfate, sulfonato-, sulfamoyl, sulfonamido-, nitro-, trifluoromethyl, cyano-, azido-, heterocyclyl, alkylaryl, or aromatic or heteroaromatic groups. Aryl groups can also be condensed or 7) bridged to alicyclic or heterocyclic rings that are not aromatic, so that . a polycycle was formed (for example, tetralin). and?" The terms "alkenyl" and "alkynyl" refer to unsaturated aliphatic groups having lengths and possible substituents analogous to the alkyls described above, but which contain at least one double or triple bond, respectively. Thus, for example, the present invention contemplates cyano and propargyl groups.-I With regard to the number of carbon atoms, the term "lower alkyl" as used herein, unless otherwise specified, means an alkyl group as defined above, but having from one to ten carbon atoms, more preferably from one to six carbon atoms on its backbone, and even more preferably from one to three carbon atoms on its backbone. Similarly, "lower alkenyl" and "lower alkynyl" have similar chain lengths. and "Alkoxyalkyl", "polyaminoalkyl" and "thioalkyloxyalkyl" refer to the alkyl groups described above which,

F also includes oxygen, nitrogen, or sulfur atoms replacing one or more carbon atoms of the hydrocarbon backbone, for example, oxygen, nitrogen, or sulfur atoms.

The terms "polycyclyl" or "polycyclic radical" mean a radical consisting of two or more cyclic rings (eg, cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, or heterocyclyl) where the two linked rings share two or more carbon atoms, for example, these rings are called "condensed

F) rings". Rings connected by non-adjacent atoms are called "bridging" rings. Each ring of this ka polycycle can be substituted with the substituents described above, such as, for example, halogen, hydroxyl, alkylcarbonyloxy-, arylcarbonyloxy-, alkoxycarbonyloxy-, aryloxycarbonyloxy- , carboxylate, in alkylcarbonyl, alkoxycarbonyl, aminocarbonyl, alkylthiocarbonyl, alkoxy, phosphate, phosphonato-, phosphinato-, cyano-, amino- (including alkylamino-, dialkylamino-, arylamino-, diarylamino- and alkylarylamino), acylamino- (incl. alkylcarbonylamino-, arylcarbonylamino-, carbamoyl and ureido-), amidino-, imino-. , 65 alkylaryl, or aromatic or heteroaromatic groups.)

The term "heteroatom" as used means an atom of any element other than carbon or hydrogen.

The preferred heteroatoms are nitrogen, oxygen, sulfur and phosphorus atoms.

The term "amino acids" means natural and unnatural amino acids found in proteins, for example, glycine, alanine, valine, cysteine, leucine, isoleucine, serine, threonine, methionine, glutamic acid, aspartic acid

Acid, glutamine, asparagine, lysine, arginine, proline, histidine, phenylalanine, tyrosine and tryptophan.

Amino acid analogues are amino acids with elongated or shortened side chains or with modified side chains with appropriate functional groups. Amino acids are also UO- and | - stereoisomers of an amino acid, if the structure of this amino acid allows the formation of such stereoisomeric forms. The term "dipeptide" means two or more 70 amino acids attached to each other. Preferred dipeptides are two amino acids connected by a peptide bond. Particularly preferred dipeptides are, for example, alanine-alanine and glycine-alanine.

It should be noted that the structure of some compounds of the present invention includes asymmetric carbon atoms.

It should also be noted that the scope of this invention, unless otherwise specified, includes isomers formed as a result of such asymmetry (for example, all enantiomers and diastereomers). Such isomers can be obtained mainly in pure form by classical methods of separation and stereochemically regulated synthesis.

The present invention relates to pharmaceutical compositions for the treatment of conditions corresponding to

M-6-substituted 7-deazapurine in mammals, for example, respiratory diseases (eg, asthma, bronchitis, chronic obstructive pulmonary disease and allergic rhinitis), renal diseases, gastrointestinal diseases and eye diseases. This pharmaceutical composition includes a therapeutically effective amount

of M-6-substituted 7-deazapurine described above and a pharmaceutically acceptable carrier. It should be noted that all deazapurines described above are intended for therapeutic treatment. In addition, it should be noted that the deazapurines of the present invention can be used either alone or in combination with other deazapurines of the present invention, or in combination with auxiliary therapeutic compounds such as, for example, antibiotics, anti-inflammatory agents or anti-cancer agents.

The term "antibiotic" is understood by a person skilled in the art and means substances produced by the cultivation of i) microorganisms and their synthetic derivatives, which prevent or inhibit the growth of pathogens and have selective toxicity in relation to pathogens, but which, at the same time, minimally affect or do not adversely affect the body of the infected host. Suitable examples of antibiotics include, but are not limited to, basic He

Zo Classes of aminoglycosides, cephalosporins, chloramphenicols, fusidic acids, macrolides, penicillins, polymyxins, tetracyclines and streptomycins. at

The term "anti-inflammatory agent" is understood by specialists and means agents acting on certain mechanisms in the body of the host without any direct antagonistic effect on the etiological factor of inflammation, such as glucocorticoids, aspirin, ibuprofen, NSAIDs, etc. with

The term "anticancer agent" is understood by a person skilled in the art and means agents that contribute to the reduction or destruction of a tumor or the prevention of the growth of cancer cells, preferably without adversely affecting other physiological functions. Typical examples are cisplatin and cyclophosphamide.

If the compounds of this invention are administered in the form of pharmaceutical preparations to a human or mammal, they may be administered as such or in the form. a pharmaceutical composition containing, for example, 0.1-99.5965 " (more preferably, 0.5-9095) of the active ingredient in combination with a pharmaceutically acceptable carrier. The term "pharmaceutically acceptable carrier" as used herein means pharmaceutically acceptable

And a material, composition, or carrier, such as a liquid or solid filler, diluent, carrier, solvent, or encapsulating material, that provides delivery or transport of the compound(s) of this invention to a given site or to a given region of the body so that it can perform the desired function. Basically, such compounds are transferred or transported from one organ or part of the body to another organ or part of the body. -I Each carrier must be "acceptable" in the sense that it must be compatible with the other ingredients of the given composition and must not adversely affect the patient. Some examples of materials that can serve as pharmaceutically acceptable carriers are sugars such as lactose, glucose, and sucrose; starches such as corn starch and potato starch; cellulose and its derivatives, such as sodium-containing carboxymethyl cellulose, ethyl cellulose and cellulose acetate; powdered tragacanth; malt; gelatin; talc; and fillers such as cocoa butter and wax for suppositories; oils such as peanut oil, seed oil

Cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; glycols such as propylene glycol; polyols such as glycerol, sorbitol, mannitol and polyethylene glycol; an ester such as ethyl oleate and ethyl laurate; agar; buffering agents such as magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogenic water; isotonic physiological solution; Ringer's solution; ethanol; phosphate-buffered solutions and other non-toxic compatible substances used in pharmaceutical compositions. (F, As described above, some embodiments of the compounds of this invention may include a basic functional group such as amino or alkylamino, and are therefore capable of forming pharmaceutically acceptable salts with pharmaceutically acceptable acids. The term "pharmaceutically acceptable salts" in this context means relatively non-toxic because the addition salts of the compounds of this invention are formed by inorganic or organic acids. These salts can be obtained either in the final isolation and purification of the compounds of this invention, or by separate reaction of the purified compound of this invention in its free basic form with a suitable organic or inorganic acid and isolation of the salt thus obtained Typical salts are hydrobromide, hydrochloride, sulfate, bisulfate, phosphate, nitrate, acetate, valerate, oleate, palmitate, stearate, laurate, benzoate, 65 lactate, phosphate, tosylate, citrate, maleate, fumarate, succinate , tartrate, naphthylate, mesylate, glucoheptonate, lactobionate and lauryl sulfonate, etc. | see, for example, B where are you? (1977) "Rpaptaseciisa! Zaiv", 9U.RNnagt.

si. 66:1-19).

In other cases, the compounds of this invention may contain one or more acidic functional groups and thus have the ability to form pharmaceutically acceptable salts with pharmaceutically acceptable bases. In these cases, the term "pharmaceutically acceptable salts" means relatively non-toxic addition salts of the compounds of this invention, formed with inorganic or organic bases. These salts can be obtained directly in the final isolation and purification of the compounds of this invention, or by separate reaction of the purified compound of this invention in its free acid form with a suitable base, such as the hydroxide, carbonate or bicarbonate of a pharmaceutically acceptable metal cation, with ammonia or with a 7/0 pharmaceutically acceptable organic primary, secondary or tertiary amine. Typical salts of alkaline or alkaline earth metals are salts of lithium, sodium, potassium, calcium, magnesium, aluminum, etc. Typical organic amines that can be used to form basic addition salts are ethylamine, diethylamine, ethylenediamine, ethanolamine, diethanolamine, piperazine, etc.

The term "pharmaceutically acceptable ester" means the relatively non-toxic esterified products of the 7/5 bpoluk of this invention. This complex ester can be obtained either in the process of the final isolation and purification of the compounds of this invention, or by means of a separate reaction of the purified compound of this invention in its free acid form or in the hydroxyl form with a suitable esterifying agent. Carboxylic acids can be converted into esters by treatment with alcohol in the presence of a catalyst. Hydroxyl trimer derivatives can be converted into an ester by treatment with an esterifying agent such as 20 alkanoyl halides. The term also refers to lower hydrocarbon groups capable of solvation under physiological conditions, such as alkyl, methyl, ethyl, and propyl esters (see, e.g., Wegode et al., supra).

In addition, the present invention contemplates the use of prodrugs that are converted into therapeutic compounds of the present invention | see, for example, K.V. 5iimeg-tap, 1992, "Tpe Ogdapis Spetivigu oi YuOgad sch ov Bevidp apa Ogyd Asiop", Asadetis Rgez5, Spr.8|. Such prodrugs can be used to modify the biodistribution (for example, of compounds that should not normally enter the protease reaction site) or i) the pharmacokinetics of a therapeutic compound. So, for example, a carboxylic acid group can be esterified, for example, with a methyl group or an ethyl group to form an ester. When this ester is administered to an individual, said ester undergoes enzymatic or non-enzymatic, heso-reductive or hydrolytic cleavage with the formation of an anionic group. The anionic group can be esterified with groups (for example, acyloxymethyl ester) desorbed to form an intermediate compound, which then decomposes to form an active compound. In another embodiment of M of the invention, these prodrugs are a reduced form of a sulfate or sulfonate, for example, a thiol, which is oxidized immediately with the formation of a therapeutic compound. In addition, the anionic group can be esterified with c with the formation of a group that is actively transported by the liver, or that is selectively absorbed by target organs. This complex ester can be chosen so that it provides specific delivery of therapeutic molecules to specific reaction sites described below for carrier molecules.

These compositions may also contain wetting agents, emulsifiers and lubricants, such as sodium lauryl sulfate and magnesium stearate, as well as dyes, releasing agents, coating agents, sweeteners, fragrances and flavors, preservatives and antioxidants. Examples of pharmaceutically acceptable antioxidants are: water-soluble antioxidants such as ascorbic acid. acid, cysteine hydrochloride, sodium bisulfate, sodium metabisulfate, sodium sulfite, etc.; soluble in and? antioxidant oils, such as ascorbyl palmitate, butylated hydroxyanisole (BEB), butylated hydroxytoluene (BHT), lecithin, propyl gallates, alphytocopherol, etc.; and agents that create chelating complexes with metals, such as citric acid, ethylenediaminetetraacetic acid (EOTA), sorbitol, tartaric acid, phosphoric acid, and the like.

Compositions of this invention are compositions suitable for oral, nasal, local, transdermal, transbuccal, sublingual, rectal, vaginal and/or parenteral administration. These -I compositions can, in general, be present in the form of a single dosage form and can be obtained by any methods well known to specialist pharmacists. The amount of the active ingredient that can be combined with the carrier material to obtain a single dose is usually the amount of the compound that is necessary for

F achieving a therapeutic effect. Generally, based on the weight of the entire composition of 10,095, this amount is from about 195 to about 9,995 active ingredients, preferably from about 595 to about 70,905, and most preferably from about 1,095 to about 3,095 active ingredients.

Methods of obtaining these pharmaceutical preparations or compositions include the stages of combining the compound of this invention with the specified carrier and, optionally, with one or more additional ingredients. IN

F) basically, these compositions are obtained by homogeneous and uniform mixing of the compounds of this invention with liquid carriers or with finely dispersed solid carriers, or with both carriers, and, if necessary, with subsequent giving of the desired shape to this product. The compositions of the present invention suitable for oral administration may be prepared in the form of capsules, wafers, dragees, tablets, lozenges (using a flavoring base, of course, sucrose, gum arabic or tragacanth), powders, granules, or in the form of a solution or suspensions in aqueous and anhydrous liquids, or in the form of an oil-in-water or water-in-oil liquid emulsion, or in the form of an elixir or syrup, or in the form of pastilles (using an inert base such as gelatin and glycerin or sucrose and 65 gum arabic) and/or in the form of a mouthwash, etc., each of which contains a predetermined amount of the compound of this invention as an active ingredient. The compound of this invention can also be administered in the form of a bolus, medicated slurry or paste.

In solid dosage forms of the present invention for oral administration (capsules, tablets, pills, dragees, powders, granules, etc.), the active ingredient is mixed with one or more pharmaceutically acceptable carriers, such as sodium citrate or dicalcium phosphate, and/or with any of the following carriers or additives, such as starch, lactose, sucrose, glucose, mannitol and/or silicic acid; binders such as, for example, carboxymethylcellulose, alginates, gelatin, polyvinyl, pyrrolidone, sucrose and/or gum arabic; humectants such as glycerin; disintegrating agents such as agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, some silicates and sodium carbonate/dissolution inhibitors, such as paraffin; absorption accelerators such as quaternary ammonium compounds; wetting agents such as, for example, cetyl alcohol and glycerol monostearate; absorbents such as kaolin and bentonite clay; lubricants such as talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate and their mixtures; and dyes. In the case of capsules, tablets, etc., pharmaceutical compositions may also contain buffering agents. Solid compositions of a similar type can also be introduced as fillers in soft and /5 Hard gelatin capsules using carriers such as lactose or milk sugar, as well as high molecular weight polyethylene glycols, etc.

Tablets can be made by compression or molding, optionally with one or more auxiliary ingredients. Compressed tablets may be prepared using a binder (eg, gelatin or hydroxypropyl methylcellulose), a lubricant, an inert solvent, a preservative, a disintegrating agent (eg, sodium-retaining starch glycolate or structured sodium-retaining carboxymethylcellulose), a surfactant, or a dispersing agent. agent

Molded tablets can be made by molding a mixture of a powdered compound moistened with an inert liquid solvent in a suitable device.

Tablets and other solid dosage forms of the pharmaceutical compositions of the present invention, such as dragees, capsules, tablets, and granules, may be, but are not required to be scored, or may be coated and coated, such as solubilized coatings and other coatings well known to pharmaceutical specialists. For i) slow or controlled release of the active ingredient of this invention, these compositions can also be made, for example, using hydroxypropyl methylcellulose in different ratios to obtain the desired release profile, other polymer matrices, liposomes and/or microspheres. They can be sterilized, for example, by filtering through a bacteria-retaining filter, or by introducing sterilizing agents in the form of sterile solid compositions, which, immediately before their use, can be dissolved in sterile water or in some other sterile medium for other actions These M compositions may also, but not necessarily, contain opacifying agents, and they may be formulated as compositions that release the active ingredient(s) only, or preferentially, in a specific area of the gastrointestinal tract. , not necessarily in extended mode. Examples of encapsulated cha compositions that can be used are polymeric substances and wax. The active ingredient may also be present in microencapsulated form, if necessary, in combination with one or more of the above-mentioned excipients.

Liquid dosage forms for oral administration of the compounds of this invention are pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups and elixirs. In addition to the active ingredient, liquid She)s dosage forms may contain inert solvents that are commonly used by specialists, such as for example, water and other solvents that solubilize agents and emulsifiers, such as ethyl alcohol, and? isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, oils (including cottonseed oil, peanut oil, corn oil, safflower oil, olive oil, castor oil, and sesame oil), glycerol, tetrahydrofuryl alcohol, polyethylene glycol-I and ester of fatty acids and sorbitan, and their mixtures.

In addition to inert solvents, oral compositions may also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening agents, flavoring agents, coloring agents, fragrances, and preservatives.

Suspensions, in addition to active compounds, may contain suspending agents, such as, for example, ethoxylated o isostearyl alcohols, polyoxyethylene sorbitol and sorbitan ester, microcrystalline cellulose,

F aluminum metahydroxide, bentonite, agar-agar and tragacanth and their mixtures.

Preparations of pharmaceutical compositions of this invention for rectal or vaginal administration can be made in the form of suppositories, which can be obtained by mixing one or more compounds of this invention with one or more suitable non-irritating fillers or carriers containing, for example, cocoa butter, polyethylene glycol, wax for suppositories or salicylate, and which are solid at room temperature

F) temperature, and liquid at body temperature, and therefore they will melt in the rectum and in the vagina with the release of the active compound.

Preparations of this invention, suitable for vaginal administration, are also pessaries, tampons, creams, gels, pastes, foams or sprays containing the indicated carriers, and such preparations are well known to those skilled in the art.

Dosage forms for local or transdermal administration of the compound of this invention are powders, sprays, ointments, pastes, creams, lotions, gels, solutions, plasters and inhalers. The active compound may be admixed under sterile conditions with a pharmaceutically acceptable carrier and with any necessary preservatives, buffers or propellants. 65 Ointments, pastes, creams and gels, in addition to the active compound of this invention, may contain fillers, such as animal and vegetable fats, oils, wax, paraffins, starch, tragacanth gum, cellulose derivatives,

polyethylene glycols, silicones, bentonites, silicic acid, talc and zinc oxide or their mixtures.

Powders and sprays, in addition to the compounds of this invention, may contain fillers such as lactose, talc, silicic acid, aluminum hydroxide, calcium silicates and polyamide powder or mixtures of these substances. Sprays

May also contain standard propellants such as chlorofluorocarbons and volatile unsubstituted hydrocarbons such as butane and propane.

Patches for transdermal administration have the additional advantage of providing controlled delivery of the compound of this invention into the body. Such dosage forms can be prepared by dissolving or dispersing this compound in a suitable medium. Absorption enhancers can also be used to increase the 7/0 rate of passage of this compound through the skin.

The rate of such passage can be regulated either by using a rate-regulating membrane or by dispersing the active compound in a polymer matrix or gel.

Ophthalmic preparations, eye ointments, powders, solutions, etc. are also included in the scope of this invention. Such a pharmaceutical preparation is preferably an ophthalmic preparative form (for example, a preparative form for injections into the peri-ocular, retrobulbar or intraocular region, a preparative form for systemic administration or a surgical solution for washing).

Ophthalmic compositions of this invention may include one or more deazapurines and a pharmaceutically acceptable carrier. At the same time, different types of media can be used. These media, by their nature, are usually aqueous. Basically, aqueous solutions obtained on the basis of the composition are preferred, as well as with the calculation that these compositions can be easily instilled into the patient's affected eyes one or two drops at a time.

However, deazapurins of this invention can also be easily introduced into compositions of other types, such as suspensions, viscous or semi-viscous gels, or solid or semi-solid compositions of other types. The ophthalmic compositions of the present invention may also include other various ingredients such as buffers, preservatives, co-solvents and viscosity-enhancing agents. high school

Appropriate buffer systems (for example, sodium phosphate, sodium acetate, or sodium borate) can be added to prevent pH changes during storage. and)

Ophthalmic products are usually packaged in reusable packaging. Therefore, to prevent microbial contamination during use, it is necessary to add preservatives. Suitable preservatives are benzalkonium chloride, thimerosal, chlorobutanol, methylparaben, propylparaben, phenylethyl (o-zo) alcohol, disodium edetate, sorbic acid, polyquaternium-1 or other agents known to those skilled in the art. Such preservatives are usually used in an amount of about 0.001 to 1.095 wt./vol .(95 "mass./vol."). me)

If the deazapurines of this invention are administered during intraocular surgery, such as retrobulbar or periocular injection and intraocular perfusion or injection, the most preferred carriers are balanced saline solutions for irrigation. Examples of balanced physiological solutions for washing are sterile washing solutions B55FO and sterile washing solutions B55 Ри for intraocular administration (AiYisop I Aroghaigiev, Ips., Gogi MMopii, Tech. O5A).

The last of these types of solutions is described in the US patent Mo4550022 (Sagabreadiap, ei ai.), the full content of which is included in this description as a reference. Injections into the retrobulbar and periocular region are known to every expert and are described in many publications, including, for example. Orpiiaitis of Zygdegu: Rgisiriez oi Rgasiise, Ea., O.I. « 0 Zraeip. M.V. Zapaege So., RpPadei!rnia, Ra., OZA, radev 85-87 (1990). with c As indicated above, the use of deazapurines to prevent or reduce retinal and optic disc tissue damage at the cellular level is the most important aspect of one of the variants of this invention. Ocular diseases that may be treated include, but are not limited to, retinopathy, macular degeneration, ocular ischemia, glaucoma, and lesions associated with ocular tissue damage such as ischemia reperfusion damage, photochemical damage, and damage associated with with -And surgery of the eye, and in particular, damage to the retina or optic nerve disc under the influence of light radiation or surgical instruments. These compounds can also be used as an adjuvant for eye surgery, such as injection into the vitreous body or into the subconjunctiva of the eye after eye surgery. These compounds can be -And used for the express treatment of fleeting conditions, or they can be administered continuously in degenerative diseases. These compounds can also be used prophylactically, and in particular, before eye surgery or non-invasive ophthalmic procedures or surgery of other

F types.

Pharmaceutical compositions of this invention suitable for parenteral administration contain one or more compounds of this invention in combination with one or more pharmaceutically acceptable sterile 5v isotonic aqueous or anhydrous solutions, dispersions, suspensions or emulsions, or with sterile powders which, immediately before their use , can be converted into sterile solutions or

F) dispersions for injections, and which may contain antioxidants, buffers, bacteriostats, dissolved substances that make them isotonic with respect to the blood of a given recipient, or suspending or thickening agents.

Examples of suitable aqueous or anhydrous carriers that can be used in the pharmaceutical compositions of this invention are water, ethanol, polyols (such as glycerin, propylene glycol, polyethylene glycol, etc.) and their respective mixtures, vegetable oils such as olive oil, and an injectable organic ester such as ethyl oleate. At the same time, appropriate fluidity can be maintained, for example, by using coating materials such as lecithin, by maintaining the desired particle size in the case of dispersions, and by using surfactants.

These compositions may also contain adjuvants such as preservatives, wetting agents, emulsifying agents and dispersing agents. To prevent the action of microorganisms, various antibacterial and antifungal agents can be included in the composition, for example, paraben, chlorobutanol, phenolsorbic acid, etc. It may also be desirable to include in these compositions isotonic agents, such as sugar, sodium chloride, etc.

In addition, prolonged absorption of the injected pharmaceutical form can be achieved due to the inclusion of agents that provide delayed absorption, such as aluminum monostearate and gelatin.

In some cases, to achieve a prolonged effect of the drug, it is desirable to slow down the absorption of the drug after subcutaneous or intramuscular injection. This can be accomplished by using a liquid suspension of a crystalline or amorphous material that has poor solubility in water. Therefore, 7/0 the rate of absorption of the drug depends on the rate of its dissolution, which, in turn, may depend on the size of the crystals and the crystalline form. Alternatively, delayed absorption of the parenteral dosage form is achieved by dissolving or suspending the drug in an oil carrier.

Depot-forms for injections are made by forming matrices that are introduced into microcapsules, in which these compounds are included in biodegradable polymers, such as polylactide-polyglycolide. The rate of release of the drug can be adjusted depending on the ratio of the amount of the drug to the amount of polymer and the nature of the polymer that is specifically used. Examples of other biodegradable polymers are poly(orthoethers) and poly(anhydrides). Compositions for injections in the form of depot forms are also made by introducing the drug into liposomes or microemulsions compatible with body tissues.

The preparations of this invention can be administered orally, parenterally, topically or rectally.

Of course, they must be made in a form suitable for each method of administration. So, for example, they can be administered in the form of tablets or capsules by injection, inhalation, eye drops or ointment, or in the form of suppositories by rectal administration. At the same time, oral administration is preferred. with

The terms "parenteral administration" and "administered parenterally" as used herein refer to other methods of administration other than intravenous or topical administration, and of course, they mean methods of i) administration by injection including, but not limited to, intravenous, intravenous Ulcerative, intraarterial, intramembranous, intracapsular, intraocular, intracardiac, transcutaneous, intraperitoneal, transtracheal, subcutaneous, subcuticular (through the subcutaneous layer), Gezo Intraarticular, subcapsular, subarachnoid, intraspinal and intrathoracic injection and infusion.

The terms "systemic administration," "administered systemically," "peripherally administered," and "peripherally administered" as used herein refer to such administration of a compound, drug, or other material in which no Kk. there is a direct delivery to the central nervous system, but in which this drug enters the patient's body, and thereby undergoes metabolism and other similar processes, for example, subcutaneous administration. with

These compounds can be administered to humans and other animals subject to treatment by any appropriate route of administration, including oral; intranasal, for example, in the form of a spray; rectal; vaginal; parenteral; intracisternal or local administration, for example in the form of powders, ointments or drops, including transbuccal and sublingual administration.

Regardless of the chosen method of treatment, the compounds of the present invention, which can be used in the appropriate hydrated form, and/or the pharmaceutical compositions of the present invention are prepared in the form of pharmaceutically acceptable dosage forms by standard methods known to those skilled in the art.

Specific doses of the active ingredients in the pharmaceutical compositions of this invention can vary depending on the effective amount of the active ingredient required to achieve the desired therapeutic response in a given patient, on the composition and on the method of administration, and do not have a toxic effect on the patient.

The dose selected will depend on a number of factors, including the activity of the particular compound of the present invention being used or its ester, salt or amide, the route of administration, the time of administration, the rate of release of the particular compound being used, the duration of treatment, other drugs, compounds where and/or materials used in combination with this compound, age, sex, weight, disease, general condition and medical history of the patient being treated, and other factors well known to those skilled in the art.

The average specialist, doctor or veterinarian, can easily determine and prescribe the effective amount of the required pharmaceutical composition. So, for example, a doctor or veterinarian may start with lower doses of a given compound

F of the invention, present in this pharmaceutical composition, as needed to achieve the desired therapeutic effect, and then gradually increase the dose until the desired effect is achieved.

Basically, an appropriate daily dose of a compound of the present invention may be the amount of the compound that is the lowest dose effective for producing a therapeutic effect. Such an effective dose will mainly depend on the factors described above. Basically, doses of the compounds of this invention for (F) intravenous and subcutaneous administration to a patient range from about 0.0001 to about 200 Ωg per kilogram of body weight per day, more preferably from about 0.01 to about 150 mg per kilogram of body weight per day, and even more preferably, from about 0.02 to about 14Omg per kilogram of body weight per day. If necessary, the effective daily dose of the active compound can be administered in the form of two, three, four smaller doses separately at appropriate time intervals during the day, and optionally in the form of single dosage forms.

Although the compound of this invention may be administered alone, it is preferred that it be administered in the form of a pharmaceutical composition. 65 The present invention also relates to packaged pharmaceutical compositions for the treatment of a condition responsive to M-6-substituted 7-deazapurine, for example, an unwanted increase in adenosine receptor activity in a mammal. These packaged pharmaceutical compositions include a container containing a therapeutically effective amount of at least one deazapurine described above and instructions for administering said deazapurine to treat a deazapurine-responsive condition in a mammal.

Deazapurins of this invention can be obtained by standard methods of organic synthesis.

Deazapurines can be purified using reversed-phase HPLC, chromatography, recrystallization, etc., and their structures confirmed by mass spectral analysis, elemental analysis, IR and/or NMR spectroscopy.

Usually, the synthesis of intermediate compounds, as well as deazapurines of this invention, is carried out in solution. The addition and removal of one or more protecting groups is also common practice and well known to those skilled in the art. Typical 7/0 synthesis schemes for obtaining the deazapurine intermediates of this invention are described below in Scheme I.

The present invention is further illustrated by the following examples, which should not be construed as limiting the present invention. The contents of all works considered for patent applications and published patent applications cited in this application, including those works that are indicated in the section "Prerequisites for the creation of the invention, are incorporated into this description by reference. At the same time, it should be noted that the models that /5 Are used in the examples, are generally accepted models, and that the illustration of efficacy using these models is predictive of human efficacy.

Deazapurins of this invention can be obtained by standard methods of organic synthesis.

Deazapurines can be purified by reversed-phase HPLC, chromatography, recrystallization, etc., and their structures can be confirmed by mass spectral analysis, elemental analysis, IR and/or

NMR spectroscopy.

Of course, the synthesis of intermediate compounds, as well as deazapurines of this invention, is carried out in solution. Addition and removal of one or more protecting groups are also common procedures and well known to those skilled in the art. Typical synthesis schemes for obtaining deazapurine intermediates of this invention are described below in Scheme I. and Scheme | oh and and sch ke re

ХХ Шле ХИ and NL V. Pyridin, SNSC in V

AH X-halide or

Me РЬ He) 30 . . about co

Meon, H2504 kr. PRA -

Om X pos s me" ! - o y! ROSIZ Y ke "

NK x -- -ya M U

АХ, И о-Ж Ж Du in 1050 S ak - s de Ku, Kv and Ko are defined above. :c» Basically, the protected 2-amino-3-cyano-pyrrole can be treated with an acyl halide to give carboxyamido-3-cyano-pyrrole, which can be treated with acidified methanol to close the ring with the resulting pyrrolo|2,34|pyrimidine-4 (Z3H)-onu (MiiMeg, S.E. ei aiYi, 9U.Mei4.Spet. 40:4396(1997)). Removal of the -1 15 pyrrole protecting group followed by treatment with a chlorinating reagent, for example, phosphorus oxychloride, leads to the formation of substituted or unsubstituted 4-chloro-7H-pyrrolo-I(2,3a|pyrimidines. As a result of the treatment of co-chloropyrimidine with amines, 7-deazapurines are obtained. - Thus, for example, as shown in Scheme II, /M-(1-41-phenylethyl)-2-amino-3-cyano-pyrrole is treated with an acyl halide in pyridine or dichloromethane. - (95) 20 phenylcarboxy-amido-3-cyano-pyrrole is treated with a mixture (10:1) of methanol/sulfuric acid to close the ring with

F by obtaining a1-7H-7-(1-phenylethyl)-pyrrolo(2,Za|Ipyrimidin-4(ZH)-one. As a result of the removal of the phenylethyl group by treating the pyrimidine with polyphosphoric acid (PPA), and then ROSI with, a key intermediate is obtained compound 4-chloro-7H-pyrrolo|(2,3a|pyrimidine. After treatment of 4-chloro-7H-pyrrolo|2,34|Ipyrimidine with various amines, indicated in Table 1, compounds of formulas (I) and (1) are obtained.

F) name) 60 b5

TABLYTSYAI - uter 343.2 o 35127 9 nm i pt MN 4e C

35035

She sh-I-lt M.

X bu pdv 3377 i 35544 i

M

Moss but t

Sh 36419 sa 40432

Thinner 3301 years 33675 pits M ia KO | still born

E, M t 35547!

YOU are

I: Ge) 35078 9 35341 o po kv (Se)

From 5545

M -- MN o Y co

Shi I- - I -

Zob 359.36 o-./ s chn, OVC and S cho m.

EK 37540 s" nni x " y ry 35035 o no 387 Z

Go so on m i 10333 ; 34128 and SS

HO lo M i

And - - kn 35145 that 33755 o "on No in (95) 4) ko bo b5

330.37 29522 ref

Y n ho 40723 3512

H s 355.45 МН 33753

CO o 70 "on o a 44133 o Mn 3502 o, k . ge xx 43.4 che EK o y SCHO ; 37248 3732

B ro A n 3072

S i s - y y y y U not y

The general method of obtaining 6b-substituted pyrroles is illustrated in the scheme below (Scheme I). at

Scheme II

S d that - a - pr "PU ise) zh M so n mya re be - ne la - vkh " m'ya shchi sch m o i-o i- on a vn » go. - well, you - well

Y Mk co. zh, M mya M na we Y « | - s rov i s t ru p v/h I and nn de R. - P5 defined above. As a result of transesterification and alkylation of ethyl cyanoacetate with o-halogen ketone, ketomethyl ka ether is obtained. After the ketone protection reaction followed by treatment with amidine hydrochloride (e.g., alkyl-, aryl-, or alkylarylamidine), a pyrimidine protected by conversion to a ketal is obtained. After removal

Che protective group with subsequent cyclization and treatment with phosphorus oxychloride gives a chloride intermediate

Ge) 20 compound, which can then be treated with an amine to give an amine-β6-substituted pyrrole. In addition, alkylation

Ф of pyrrole in the nitrogen atom can be achieved under conditions known to a person skilled in the art.

The general method of obtaining 5-substituted pyrroles is illustrated in the scheme below (Scheme PP).

Scheme PI name) 60 b5 o

M SM Me. Sk

Me. cm I АХ -Ж-| ri yatzo and k ve v

V i - - voi vsta / vat; "I am

Kk ii n Kk paye si yuoru -- So mch M m M,

MK, 15 | n вх Mn de Ku - Ko are defined above, and K represents a protecting group that is removed.

Condensation of malononitrile and excess ketone with subsequent bromination of this product 20 gives a mixture of starting material, monobrominated and dibrominated products, which is then treated with alkylamine, arylamine or alkylarylamine. The resulting amine product is acylated with chloride, and this monoacylated pyrrole is cyclized in the presence of acid to give the corresponding pyrimidine. The pyrrole protecting group is removed using polyphosphoric acid and treated with phosphorus oxychloride to give the chlorinated product. This chlorinated pyrrole can then be treated with an amine to give a 25% amino-5-substituted pyrrole. Alkylation of pyrrole at the nitrogen atom can be carried out under conditions known to a person skilled in the art.

Schemes IM and M illustrate methods of obtaining deazapurines 1 and 2 of this invention. o t IF: n MN t Te; ich x M Z 30 | in district (ze)

ZK IN su MN cha 1 z

And I s de K» and Ke are as they were defined above, for example, CH.

З A specific example of obtaining b-methylpyrrolopyrimidines i-

The key reaction for obtaining b-methylpyrrolopyrimidines (1) (|K5 - CH) is the reaction of cyclization of cyanoacetate with benzamidine to form pyrimidine. Obviously, methyl cyanoacetate should cyclize with benzamidine to form a pyrimidine more efficiently than the corresponding ethyl ether. Therefore, as a result of transesterification and alkylation of ethyl cyanoacetate in the presence of MaOMe and an excess of o-haloacetyl group, for example - 40 chloroacetone, the desired methyl ester (3) was obtained with a yield of 7995 (Scheme IM). Complex ketoester c (3) was protected in the form of acetal (4) with a yield of 8195. A new method of cyclization to obtain pyrimidine (5) "" was carried out with amidine hydrochloride, for example, benzamidine hydrochloride and with 2 equivalents of YUOVA, as a result of which "pyrimidine was obtained 5 with a yield of 5495. This method allows you to increase the yield from 2095 and more using the conditions described in the literature, which involve the use of MaOMe in the process of cyclization with 45 guanidine. Cyclization with the formation of pyrrolepyrimidine (6) is achieved by removing the acetal group in aqueous HC1 with a yield of 7895. As a result of the reaction of compound (6) with phosphorus oxychloride under reflux, alkylation (7) was obtained. As a result of the interaction reaction with trans-4-aminocyclohexanol in dimethylsulfoxide at 135930, compound (1) was obtained with a yield of 5795 starting from compound (7). It is obvious to everyone and a specialist that the choice of reagents gives great freedom in choosing the right deputy of Kb.

OO 250 Scheme IM son nin - "Zh - bul -80-still-Osh ko Mo v n z

F ( o» G r na Fo sh--k o o su ky sha; ; 60 and yuyu. t a Iva br. --ADD- So. s. u x M S s po na she s Y y y ,

A specific example of obtaining 5-methylpyrrolopyrimidines

Due to the condensation of Kpoemepde! of malononitrile and an excess of a ketone, for example, acetone in boiling benzene, after distillation, compound 8 was obtained with a yield of 5095. As a result of bromination of compound 8

With M-bromosuccinimide in the presence of benzoyl peroxide in chloroform, after distillation, a mixture of the starting material, mono- (9) and di-brominated products (5/90/5) (7090) was obtained. This mixture was reacted with o-methylalkylamine or o-methylarylamine, for example, o-methylbenzylamine, resulting in aminopyrrole (10). After passing through a short column of silica gel, the partially purified amine (yield 31905) was acylated with an anhydride such as benzoyl chloride to give mono-(11) and diacylated (12) pyrroles 70, which were separated by flash chromatography. After acid hydrolysis of disubstituted pyrrole (12), acylpyrrole (11) was obtained with a total yield of 2995. As a result of cyclization in the presence of concentrated sulfuric acid and DMF, compound (13) (2395) was obtained, which was subjected to a deprotection reaction using polyphosphoric acid to obtain the compound ( 14). After the reaction of compound (14) with phosphorus oxychloride under reflux, alkylation (15) was obtained. As a result of the interaction reaction with 75 trans-4-aminocyclohexanol in dimethylsulfoxide at 135eS, compound (2) (Kv - CH3) was obtained from compound (14) with a yield of 3095 (see scheme M). It is obvious to every specialist that the choice of reagents gives great freedom in choosing the right deputy of Kv.

Scheme M? Me. SM M MS. cm rye nori oe 8 9

Mo, -- 0 Sun - Sun But SI Friday 10 o'clock

Me. - What are you doing? p oz Te) yan on so - AK - DYU - s o Si y s 1 " | - si -- Ж Fa -08-k 2 bu y « 15 shsh s Alternative method of synthesis of Co-substituted pyrroles, for example, 5-methylpyrrolopyrimidines :

This alternative method for the synthesis of Co-substituted pyrroles, for example, 5-methylpyrrolopyrimidines, involves "c" transesterification and alkylation of ethyl cyanoacetate to obtain compound (16) (Scheme MI). As a result of condensation of compound (16) with benzamidine hydrochloride and 2 equivalents of OBO, pyrimidine (17) was obtained. Cyclization with the formation of pyrrole-pyrimidine (14) is achieved by removing the acetal group in aqueous HC1. As a result of -I reaction of compound (14) with phosphorus oxychloride under reflux, the corresponding 4-chloro derivative (15) was obtained. As a result of the interaction reaction with trans-4-aminocyclohexanol in dimethylsulfoxide at about 135, compound 2 was obtained. This technique allows reducing the number of reactions of the synthesis of the target compound (2) from 9 to 4 -I stages. At the same time, the yield increases significantly. It is also obvious to anyone skilled in the art that the choice of reagents gives great freedom in choosing the right Ka substituent.

Mami Scheme MI 42)

F) ime) 60 b5 sho; oh and oh oh oh What

M

Maybe 16

We we, he ich 0o- also sh | with gu (in;

I am "7

N si Ya

M N sa

Z n ----khyazh sya --- n 0 x M i

N N

14 15

MAYA, r th rFah v n s - y y y U ne y

The general method of obtaining des-methylpyrrole is illustrated in the scheme below (Scheme MI). at

MY scheme

HM shk and

Р Ж ----ж о: (Со) 5 к Ф : bu

I. ko Mts Ge)

E cha

Mn

HA ON n an s vu Mn, peki u "a u Ay d- Hi Ue i rota M ono nn mKV, "

NMKIV2 s y-ke so" o v s Her ;" where K.-Kz are defined above.

As a result of alkylation of alkylcyanoacetate with diethylacetal in the presence of a base, Cyanodiethylacetal was obtained, which was treated with an amidine salt to obtain a methylpyrrolopyrimidine precursor. This -I precursor was chlorinated and treated with an amine to form the target des-methylpyrrolopyrimidine shown above.

So, for example, the synthesis of compound (18) is illustrated in Scheme MI. o Scheme of MY

IN

-i ke y Me 5, bo - dno oz Y o-« yi iya me no

F TV 19

MN hi, vno ' ish sa

OK, I'm 3 years old

M s (F) rova Su u na t " p

BUT. that

M zh ge o n b 18 '

Commercially available methyl cyanoacetate was alkylated with bromoacetaldehyde diethyl acetal in the presence of potassium carbonate and Ma! with the formation of compound (19). Cyclization with the formation of pyrimidine (20) was carried out in two stages. First, the pyrimidine acetal was obtained by reacting compound (19) with benzamidine hydrochloride and two equivalents of OBO. The resulting pyrimidine acetal was subjected to a deprotection reaction without purification

Water with InHC, and the resulting aldehyde was cyclized to give pyrrolepyridine (20), which was isolated by filtration. After the reaction of compound (20) with phosphorus oxychloride under reflux, the corresponding 4-chloro-derivative (21) was obtained. As a result of the reaction of the chlorine derivative with trans-4-aminocyclohexanol in DMSO at 13523, compound (18) was obtained from compound (21).

The II-MI diagrams show that 70 functional groups can be introduced into the 5- and b-positions of the pyrrolepyrimidine ring. Various functional groups can be introduced into the 5- and b-position of compounds of formula (1) and (II) by using different starting reagents and carrying out a small modification in the above-mentioned reaction schemes. Some examples are presented in Table 2.

Table 2.

Selected list of 5- and 6-substituted pyrrolepyrimidines zh (veinninntya 5 (6 ev Н N vysh 9 Х Н Substituted Ak b 07 0 SOSNY SNZ poshi shy ; с 0 o С(О)МНСНЗ | СН o пил шыШ si

The present invention is further illustrated by the following examples, which should not be construed as limiting the present invention. Contents of all pending works, patent applications and published patent applications cited in this application, including works that are listed in the section "Prerequisites for the creation of the invention", and. incorporated herein by reference. At the same time, it should be noted that the models used in these examples are well-known models, and that the illustration of effectiveness on these models allows predicting the effectiveness for humans. with

Examples of

Preparation example 1

We used a modification of the Zeeia 85 alkylation method: I irke!". To a solution of ethyl cyanoacetate (6.58 g, 58.1 mmol) cooled on ice (02) in MeonN (20 ml) was slowly added a solution of MaOMe (2595 wt/vol.; 58.1 mmol) "

After 10 minutes, chloroacetone (ml; 62.8 mmol) was slowly added. After 4 hours, the solvent was removed.

The brown oil was diluted with EtOAc (100ml) and washed with HO (100ml). The organic fraction was dried, filtered and concentrated to give a brown oil (7.79g; 7990). Oil (3) (Scheme IM) was a mixture of "» products of methyl/ethyl ester (9/1), and was used without additional purification. 91 (dd, 1H, 9U-7.2, 7.0Hz, CH), 3.62 (s, ЗН, OSN"), 3.42 (dd, 1H,

U-15,0,7,1 Hz, 1 x CH"; 3.02 (dd, 1H, 9-15.0, 7.0 Hz, 1 x CH"); 2.44 (s, ZN, SN"), 1.26 (t, 9-7.1 Hz, complex ether-SNU). and 1Teeia, E 5 GorKe ), Speth.Veg. 1977,110,1462-1469. ko Preparative example 2.

We used the technique of Zeeia and Irke!. As a result of the protection reaction of ketone (3) (Scheme IM; 5.0g, 32.2mmol) with ethylene glycol (4ml, 64.4mmol) in the presence of TON (100mg), compound (4) was obtained in the form of an oil (Scheme M, (95) 50 5 .2g, 81.0) after flash chromatography (51O", 3/7 EOAc/hexane, K. 0.35). The residual content was approximately 590

F of ethyl ether: "H-NMR (200 MHz, SOCIv) 5: 4.24 (q, U-7.2 Hz, OSN"), 3.98 (s, 4H, 2 x acetal-CHo), 3, 79 (p.

ЗН, OSН»), 3.62 (dd, 1Н, 9У-7.2, 7.0 Hz, СН), 2.48 (dd, 1Н, 915.0, 71 Hz, 1 x OSН»), 2 ,32 (dd, 1H, 9U-15.0, 7.0 Hz, 1x

SNU"); 1.35 (s, ZN, SNU), 1.26 (t, 9-71 Hz, complex ether-CH3); MS(EN) (En-electrospraying): 200.1 (M 1). 1Teeia, E 5 GorKe Ou, Spet.Veg. 1977, 110, 1462-1469. o Preparative example 3:

A solution of acetal (4) (Scheme IM, 1g, 5.02mmol), benzamidine (78bmg, 5.02mmol) and OWL (1.b5ml, 10.04mmol) in dry DMF (15ml) was heated to 85923 for 15h. The mixture was diluted with HCl (30 ml) and washed with 0.5 N MMA (10 ml) and HO (20 ml). The organic fraction was dried, filtered and concentrated to give a brown oil. 60 An attempt was made to carry out flash chromatography (5IO5; 1/9 EtOAc/CH2Cl, MK-O0.35), but the material crystallized on the column. The silica gel was washed with MeonN. Fractions containing product (5) (Scheme IM) were concentrated and used without further purification (78Zmg, 54.395). "H-NMR (200 MHz, SOSI") 5: 8.24 (m, 2H, AG-H), 7.45 (m, ZH, AG-H), 5.24 (sh.s, 2H, MH ), 3.98 (s, 4H, 2 x acetal-CH»o), 3.60-3.15 (m, 2H, CH»), 1.38 in (s, 3H, CHNas); MS (EN ): 288.1 (M 1).

Preparation of compound (20) (Scheme MI): Solution of acetal (19) (4.43g, 20.bmmol)1, benzamine hydrochloride

(3.22g, 20.bmmol) and OV (6b.15ml, 41.2mmol) in dry DMF (20Oml) were heated to 85923 for fifteen hours. The mixture was diluted with 100 ml of SNI" and washed with HO (2 x 5 Oml). The organic fraction was dried, filtered and concentrated to give a dark brown oil. The dark brown oil was stirred in NaCl (10 mL) for 2 hours at room temperature. The resulting suspension was filtered to obtain the hydrochloride salt of compound (20) as a brownish solid (3.60g, 70.695); "H-NMR (200 MHz,

DMSO-jv) 6: 11.92 (s, 1H), 8.05 (m, 2H, Ag-H), 7.45 (m, ZN, Ag-H), 7.05 (s, 1H, pyrrole -N); MS(EN): 212.1 (M).

Preparation example 4:

A solution of acetal (5) (700 mg, 2.44 mmol) in aqueous HCl (40 ml) was stirred for 2 h. at room temperature 70 The obtained suspension was filtered to obtain /HeSi-salt /2-phenyl-b6-methyl-7H-pyrrolo|2,Za|pyrimidin-4(ZH)-one in the form of a brownish solid (498 mg, 78.095): " H-NMR (200 MGu,

DMSO-α) 6: 11.78 (s, 1H), 8.05 (m, 2H, Ag-H), 7.45 (m, ZH, AgH-H), 6.17 (s, 1H, pyrrole -Н), 2.25 (с, ЗН, СН»); MS(EN): 226.1 (M'1).

Preparative example 5: The modified cyclization method of Spep et al. was used. To the ice-cooled (02) solution of bromide (9) (Scheme M; 20.0g, 1O88mmol; 9095 purity) in isopropyl alcohol (bOml) was slowly added a solution of o- of methylbenzylamine (12.5 ml, 97.3 mmol). The black solution was left to warm to room temperature and stirred for 15 hours. The mixture was diluted with EtOAc (200 ml) and washed with 0.5 N Mahon (50 ml).

The organic fraction was dried, filtered and concentrated to obtain the compound in the form of black tar (19.2 g, 94965). The residue was partially purified by flash chromatography (5iO5; 4/96 Meon/cSnsi", MK 0.35) and the compounds a1-1-(1-phenylethyl)-2-amino-3-cyano-4-methyl-pyrrole were obtained as a black solid (6.38g, 3190). MS(EN): 226.1 (M"-1).

TSpep, M. Y.; Maperasy, V.5.; Muipieg, 5.M.; Vgookv5, E.; Soiipe, U. Sogtap, MH.; YuOypaivKkiv, A.V.; Ragasi, sem 2 M.5.; SZaMazspip, K.; Zsptiai, B.; Zspytsi 7, 1997, 40, 1749-1754. at

Preparative example 6:

To the solution of a1-1-(1-phenylethyl)-2-amino-3-cyano-4,5-dimethylpyrrole! (14.9g, 62.5mmol) and pyridine (10.0ml) in dichloromethane (50.0ml) was added benzoyl chloride (9.37g, 66.7mmol) at 02C. After stirring for a while at 02C, hexane (10.00 ml) was added to facilitate precipitation of the product. The solvent was removed under vacuum and the solid was removed, the substance was recrystallized Kk) from a mixture of EUN/HNO Kk) to obtain 13.9 g (6590) of α1-1-(1-phenylethyl)-2-phenylcarbonylamino-3-cyano-4,5-dimethylpyrrole, t.pl. 218-2212C; "H-NMR (200 MHz, m

SOSISCH) 6: 1.72 (s, ZN), 1.76 (d, 9U-7.3 Gu, ZN), 1.98 (s, ZN), 5.52 (sq, U-7.3 Gu , 1H), 7.14-7.54 (m, 9H), 7.68-7.72 (dc, 9-1.4 Hz, 6.9 Hz, 2H), 10.73 (s, 1H) ; MS(EN): 344.4 (M'1). with "Sheridz App. Spect. 1986, 1485-1505, -

The following compounds were obtained by a method similar to that described in Preparative example 6: a1-1-(1-Phenylethyl)-2-(3-pyridyl)carbonylamino-3-cyano-4,5-dimethylpyrrole. "H-NMR (200 MHz, SOSIv) 5 : 1.83 (d, 9-6, 8 Hz, ZN), 2.02 (s, ZN), 2.12 (s, ZN), 5.50 ( sq., 9U-6.8 Hz, 71), 7.14-7.42 (m, 5H), 8.08 (m, 2H), « 8.75 (m, ЗН); MS(EN): 345 ,2 (M"1). 70 a1-1-(1-Phenylethyl)-2-(2-furyl)carbonylamino-3-cyano-4,5-dimethylpyrrole. "H-NMR (200 MHz, SOSIV) 5: 1.84 8 s (d, 9U-7.4 Hz, ZN), 1.92 (s, ZN), 2.09 (s, ZN), 5, 49 (q, 9-7.4 Hz, 1), 6.54 (dd, 9U-1.8 Hz, 3.6 Hz, MN), :z» 7.12-7.47 (m, 7H); MS(EN): 334.2 (M"-1), 230.1. a1-1-(1-Phenylethyl)-2-(3-furyl)carbonylamino-3-cyano-4,5-dimethylpyrrole. "H-NMR (200 MHz, SOSIV) 5: 1.80 (d, 9-7 Hz, ZN), 1.89 (s, ZN), 2.05 (s, ZN), 548 (sq, U- 7 Hz, 1H), 6.59 (s, 1H), 7.12-7.40 (m, 6H), 7.93 (s, -I 1H); MS(EN): 334.1 (M" -1), 230.0. with a1-1-(1-Phenylethyl)-2-cyclopentylcarbonylamino-3-cyano-4,5-dimethylpyrrole. "H-NMR (200 MHz, SOSIv) 8: 1382 (d, 9U-7.4 Hz, ZN), 1.88 (s, ZN), 2.05 (s, ZN), 1.63-1, 85 (m, 8H), 2.63 (m, 1H), 5.43 (kv, U-7.4 Hz, 1H), 6.52

Sh- (c, 1H), 7.05-7.20 (m, 5BN); MS(EN): 336.3 (M -1). so 20 a1-1-(1-Phenylethyl)-2-(2-thienyl)carbonylamino-3-cyano-4,5-dimethylpyrrole. "H-NMR (200 MHz, SOSIi) 8: 1.82

Ф (d, 5-6, 8 Hz, ЗН), 1.96 (с, ЗН), 2.09 (с, ЗН), 5.49 (q, 9У-6.8 Hz, 1Н), 7, 05-7.55 (m, 88); MS(EN): 350.1 (M), 246.0. a1-1-(1-Phenylethyl)-2-(3-thienyl)/carbonylamino-3-cyano-4,5-dimethylpyrrole. "H-NMR (200 MHz, SOSIv) 5: 1.83 (d,

U-7.0 Hz, ЗН), 1.99 (s, ЗН), 2.12 (s, ЗН), 549 (kv, У-7.0 Hz, 71Н), 6.90 (m, 1) , 7.18-7.36 (m, 6H), 7.79 (m, 1H); MS(EN): 350.2 (M"-1), 246.1. a1-1-(1-Phenylethyl)-2-(4-fluorophenyl)carbonylamino-3-cyano-4,5-dimethylpyrrole. TH- NMR (200 MHz, about SOSI)) 5: 1.83 (d, U-7.4 Hz, ZN), 1.96 (s, ZN), 2.08 (s, ZN), 5.51 (q , U-7.4 Hz, 1H), 7.16-7.55 (m, ZN); MS(EN): 362.2 o (Me), 25841. vo a1-1-(1-Phenylethyl)- 2-(3-Fluorophenyl)carbonylamino-3-cyano-4,5-dimethylpyrrole 1H-NMR (200 MHz, SOCIv) δ: 1.83 (d, 9U-7.4 Hz, 3H), 1.97 (s, ЗН), 2.10 (s, ЗН), 5.50 (kv, U-7.4 Hz, 71Н), 7.05-7.38 (m, 7Н), 7.67-7, 74 (m, 2H); MS(EN): 362.2 (M"-1), 258.1. a1-1-(1-Phenylethyl)-2-(2-fluorophenyl)carbonylamino-3-cyano-4,5-dimethylpyrrole. H-NMR (200 MHz, SOSIv) δ: 1.85 (d, 9U-7.2 Hz, ZN), 1.94 (s, ZN), 2.11 (s, ZN), 5.50 ( sq, U-7.2 Hz, 1H), 7.12-7.35 (m, 6H), 7.53 (m, 1H), 657.77 (m, 1H), 8.13 (m, 18 ); MS(EN): 362.2 (M"-1), 258.0. a1-1-(1-Phenylethyl)-2-isopropylcarbonylamino-3-cyano-4,5-dimethylpyrrole. 1H-NMR (200 MHz , SOSIv) 8: 1,19

(d, 9-7.0 Hz, 6H), 1.82 (d, 9U-7.2 Hz, ZN), 1.88 (s, ZN), 2.06 (s, ZN), 2.46 (m, 1H), 5.39 (m, 9-7.2 Hz, МН), 6.64 (s, 1H), 7.11-7.36 (m, 5H); MO(EN): 310.2 (M), 206.1.

Acylation of a1-1-1-phenylethyl)-2-amino-3-cyano-4-methylpyrrole gave monoacylated 50 41-1-1-phenylethyl)-2-benzoylamino-3-cyano-4-dimethylpyrrole and diacylated a1 -1-(1-phenylethyl)-2-dibenzoylamino-3-cyano-4-methylpyrrole. Monoacylated pyrrole: "H-NMR (200 MHz, SOCIv) δ: 7.69 (d, 2H, 2-78 Hz, Ag-H), 7.58-7.12 (m, 8H, AgH-H), 6.18 (c, 1H, pyrrole-H), 5.52 (q, 1H, 9U-7.2 Hz, CH-SNU), 2.05 (c, ZN, pyrrole-CHa), 1.85 ( d, ZN, U-7.2 Hz, CH-CHnui); MS(EN): 330.2 (M 1); Diacylated pyrrole: "H-NMR (200 MHz,

SOSIv) 6: 7.85 (d, 2H, 97.7 Hu, Ag-H), 7.74 (a, 2H, 0-78 HZ, Ag-H), 7.52-7.20 (m, 9H, Ag-H), 7.04 (m, 2H, 70 AN), 6.21 (s, 1H, pyrrole-H), 5.52 (q, IN, U-7.2 Hz, CH-SNaU ), 1.77 (d, ZN, 9-7.2 Hz, CH-CHNa), 1.74 (s, ZN, pyrrole-CH3); MS(EN): 434.1 (M 1).

Preparative example 7:

Concentrated sulfuric acid (1.00 ml) was added to a solution of 4a1-1-(1-phenylethyl)-2-phenylcarboxyamido-3-cyano-4,5-dimethylpyrrole (1.0 g, 2.92 mmol) in methanol (10.00 ml) at 09C. The resulting mixture was heated under reflux for 15 hours and cooled to room temperature. The precipitate was filtered to obtain 0.48 g (4895) of α-5,6-dimethyl-2-phenyl-7H-7-(1-phenylethyl)pyrrolo|2,Za|pyrimidin-4(ZH)-one. /"H-NMR (200 MHz,

SOSISCH) 8: 2.02 (d, 9-74 Hz, ZN), 2.04 (s, ZN), 2.41 (s, ZN), 6.25 (sq, U-7.4 Hz, 1Н ), 7.22-7.50 (m, 9H), 8.07-8.12 (dd, 9-34 Hz, 6.8 Hz, 2H), 10.51 (s, 1H); MS(EN): 344.2 (M"-1).

The following compounds were obtained by the method described in Preparative Example 7. a1-5, b-dimethyl-2-(3-pyridyl)-7H-7-(1-phenylethyl)pyrrolo|2,3|pyrimidin-4(ZH)-one . "H-NMR (200 MHz, SOSI"v) 5: 2.03 (d, 9U-7.2 Hz, ZN), 2.08 (s, ZN), 2.42 (s, ZN), 6, 24 (q, 9-7, 2 Hz, 1H), 7.09-7.42 (m, 5H), 8.48 (m, 2H), 8.70 (m, ЗН); MS(EN): " NA-NMR (GOO MHC, p 29 SOSIZ) 5:1.98 (d, 9-7, 8 Hz, ZN), 1.99 (s, ZN), 2.37 (s, ZN), 6.12 (q, U-7.8 Hz, 1H), 6.48 (dd, U-1.8 Hz, (U 3.6 Hz, 1H), 7.17-7.55 (m, 7H), 9 .6 (c, 1H); MS(EN): 334.2 (M 1). a1-5, b-dimethyl-2-(3-furyl)-7H-7-(1-phenylethyl)pyrrolo|2, 1H-NMR (200 MHz,

SOSI)) 5: 1.99 (d, 9-7 Gu, ZN), 2.02 (s, ZN), 2.42 (s, ZN), 6.24 (q, U-7 Gu, 1) , 7.09 (s, 1H), 7.18-7.32 "0 (m, BN), 7.48 (s, 1H), 8.51 (s, 1H); MS(EN): 334.2 (M'"-1). c a1-5,6-dimethyl-2-cyclopental-7H-7-(1-phenylethyl)pyrrolo|2, Z|pyrimidine-4 (ZH) -on. "H-NMR (200 MHz,

SOSI)) 6: 1.95 (d, 9-74 Hz, ZN), 2.00 (s, ZN), 2.33 (s, ZN), 1.68-1.88 (m, 8H), 2.97 (m, t1H), 6.10 (kv, - 97.4 Hz, 1), 7.16-7.30 (m, BN), 9.29 (s, 1H); MS(EN): 336.3 (M"-1). c a1-5, b-dimethyl-2-(2-thienyl)-7H-7-(1-phenylethyl)pyrrolo|2,Za|Ipyrimidine- 4 (ZN)-on. "H-NMR (200 MHz, SOS) 5: chn 2.02 (d, 9U-7.2 Hz, ZN), 2.06 (s, ZN), 2.41 (s ) , 7.44 (q, U-4.8 Hz, 1H), 8.01 (d, U-2.8 Hz, 1H), 11.25 (s, 1); MS(EN): 350.2 (M 1). a1-5, b-dimethyl-2-(3-thienyl)-7H-7-(1-phenylethyl)pyrrolo|2,Za|pyrimidin-4 (ZH)-one. "H-NMR (200 MHz, SOSI") 5: " 2.00 (d, 9-74 Hz, ZN), 2.05 (s, ZN), 2.43 (s, ZN), 6.24 ( sq., 9U-7.4 Hz, 1H), 7.24-7.33 (m, 5H) 5 7.33-7.39 (m, 1H), 7.85 (m, 1H), 8.47 (m, 1H), 12.01 (s, 1H); MS(EN): 350.2 (M"1). o) c a1-5, b-dimethyl-2-(4-fluorophenyl)-7H-7-(1-phenylethyl)pyrrolo|2, Za|I-pyrimidin-4 (ZH)-one. "H-NMR (200 MHz, with" SOSIv) 5: 2.01 (d, 9-68 Hz, ZN), 2.05 (s, ZN), 2.42 (s, ZN), 6.26 ( sq, U-6.8 Hz, 1H), 7.12-7.36 (m, 7H), 8.23-8.30 (m, 2H), 11.82 (s, 1H); MS(EN ): 362.3 (M"-1). a1-5, b-dimethyl-2-(3-fluorophenyl)-7H-7-(1-phenylethyl)pyrrolo|2,3a|-pyrimidin-4 (ZH)-one. "H-NMR (200 MGu, -1 SOSIv) 5: 2.02 (d, 9-74 Hu, ZN), 2.06 (s, ZN), 2.44 (s, ZN), 6.29 ( sq, U-7.4 Hz, 1H), 7.13-7.51 (m, 5H), 8.00-8.04 (m, 2H), 11.72 (s, 2H); MS(EN ): 362.2 (M'"-1). o a1-5, b-dimethyl-2-(2-fluorophenyl)-7H-7-(1-phenylethyl)pyrrolo (2,341-pyrimidin-4 (ZH)-one. "H-NMR (200 MHz, -I SOSIv ) 8: 2.00 (d, 9U-7.2 Hz, ZN), 2.05 (s, ZN), 2.38 (s, ZN), 6.24 (sq, 9U-7, 2 Hz, 1H), 7.18-7.45 (m, 8H), at 50 8.21 (m, 1H), 9.54 (s, 1H); MS(EN): 362.2 (M'-1) . a1-5,6-dimethyl-2-isopropyl-7H-7-(1-phenylethyl)pyrrolo|2,Za|pyrimidin-4 (ZH)-one. "H-NMR (200 MHz, SOCIv) 5: s 1.30 (d, 9U-6.8 Hz, ZN), 1.32 (sq, 9U-7.0 Hz, 71H), 2.01 (s, ZN), 2.34 (s, ZN), 2.90 (m, 1H), 6.13 (m, 1H), 7.17-7.34 (m, 5H), 10.16 (s, 13); MS(EN): 310.2 (M "1).

Preparative example 8: 59 Solution of a1-1-(1-phenylethyl)-2-benzoylamino-3-cyano-4-dimethylpyrrole (785mg, 2.3v8mmol) with concentrated

HF) with sulfuric acid (ml) in DMF (1 ml) was stirred at 1302 for 48 h. The black solution was diluted with CHCl3 (100 ml) and washed with 1 N Mahon (30 ml) and saline (30 ml). The organic fraction was dried, filtered, concentrated and purified using flash chromatography (5iO5, 8/2 EtOAc/hex, CI 0.35) to obtain a1-5-methyl-2-phenyl-7H-7-(1-phenylethyl) )pyrrolo|2, Za|Ipyrimidin-4(ZN)-one as a brown solid (184mg, 24965). "H-NMR (2000 MHz, SOSIv) 5: 8.18 (m, 2H, Ag-H), 7.62-7.44 (m, ZH, Ag-H), 7.40-7.18 ( m, 5H,

AgH-H), 6.48 (s, 1H, pyrrole-H), 6.28 (sq, 1H, 9U-7.2 Hz, CH-CH»U), 2.18 (s, ЗН, pyrrole- СН"), 2.07 (d, ЗН, 9-7.2 Hz,

CH-CH»U); MS(EN): 330.2 (M 7-1).

Preparative example 9: 65 A mixture of α1-1-(1-phenylethyl)-2-amino-3-cyano-4,5-dimethylpyrrole (9.60g, 40.00mmol) and formic acid (50.0ml, 9895) was heated with under reflux for 5 hours. After cooling to room temperature and scraping the walls of the flask, a large precipitate formed, which was then filtered. The product was washed with water until the pH of the wash became neutral, as a result of which a1-5,6-dimethyl-7H-7-(1-phenylethyl)pyrrolo|2,ZaIpyrimidin-4(ZH)-one was obtained. NMR (200 MHz, SOSI) 8: 1.96 (d, 9-74

HC, ЗН), 2.00 (s, ЗН), 2.38 (s, ЗН), 6.21 (kv, 9U-7.4 Hz, 1Н), 7.11-7.35 (m, 5Н ), 7.81 (c, 1H), 11.71 (c, 1H),

MS(EN): 268.2 (M'1).

Preparative example 10: a1-5,6-Dimethyl-2-phenyl-7H-7-(1-phenylethyl)pyrrolo|2,Za|pyrimidin-4(ZH)-one (1.0g, 2.9mmol) was suspended in polyphosphoric acid (3O,Oml). The mixture was heated at 1002C for 4 hours. The hot suspension was poured into water 70 with ice, intensively stirred to disperse it and alkalinized to pH 6 by adding solid KOH.

The resulting solid was filtered and collected (Kk) to obtain 0.49 g (6990) of 5,6-dimethyl-2-phenyl-7H-pyrrolo|2,Za|pyrimidin-4(ZH)-one. "H-NMR (200 MHz, DMSO-yv) δ: 2.17 (s, ЗН), 2.22 (s, ЗН), 7.45 (lat.s, З2Н), 8.07 (lat., 2H), 11.49 (s, 1H), 11.82 (s, 1H); MS(EN): 344.2 (M"1). /5 The following compounds were obtained by the method described in Preparative example 10: 5-Methyl-2-phenyl-7H-pyrrolo|2,Za|pyrimidin-4(ZH)-one. MS(EN): 226.0 (M 1). 5-Methyl-2-(3-pyridyl)-7H-pyrrolo(2,Za-pyrimidine-4(ZH)-one. MS(EN): 241.1 (M-1). 5,6-Dimethyl-2-( 2-Furyl)-7H-pyrrolo|2,ZaIpyrimidine-4(3H)-one.i "H NMR (200 MHz, DMSO-iv) δ: 2.13 (s, ЗН), 2.18 (s, ЗН ), 6.39 (dd, uU- 1.8, 3.6 Hz, 1H), 6.65 (dd, U-1.8 Hz, 3.6 Hz, 1H), 7.85 (dd, 9U -1.8, 3.6 Hz, 1H), 11.45 (s, 1H), 11.60 (s, 13); MS(EN): 230.1 (M'1). 5,6-Dimethyl -2-(3-Furyl)-7H-pyrrolo|2,ZaIpyrimidine-4(3H)-one.i "H-NMR (200 MHz, DMSO-iv) 85: 2.14 (s, ЗН), 2, 19 (s, ZN), 6.66 (s, 1H), 7.78 (s, 1H), 8.35 (s, 1H), 11.3 (s, 1H), 11.4 (s, 1H ); MS(EN): 230.1 (M"-1). 5,6-Dimethyl-2-cyclopentyl-7H-pyrrolo|2,Za|pyrimidin-4(ZH)-one.. "H-NMR ( 200 MHz, DMSO-ydv) 8: 1.57-1.91 (m, ov 8Н), 2.12 (s, ЗН), 2.16 (s, ЗН), 2/99 (m, 1Н), 11, 24 (s, 1H), 11.98 (s, 1H); MI(EN): 232.2 (M). s 5,6-Dimethyl-2-(2-thienyl)-7H-pyrrolo|2, ZaIpyrimidine-4(3ZH)-on.y "H NMR (200 MHz, DMSO-yv) δ: 2.14 (s, ЗН), о 2.19 (s, ЗН), 7.14 (dd, 9U- 3.0, 5.2 Hz, 71), 7.70 (d, 9U-5.2 Hz, 1n), 8.10 (d, 9U-3.0 GuC/ 1H), 11.50 (s, 1H);

MS(EN): 246.1 (M"1). 5, 6-Dimethyl-2-(3-thienyl,)-7H-pyrrolo|2,Za|pyrimidin-4(33H)-one. "H-NMR (200 MHz, DMSO-yv) 5: 2.17 (s, ЗН), Ф 2.21 (s, ЗН), 7.66 (m, 1Н), 7.75 (m, 1Н), 8.43 (m, 1H), 11.47 (s, 1H), 11.69 (s, 1H); MS(EN): 246.1 (M"--1). so 5,6-Dimethyl-2-(4-fluorophenyl)-7H-pyrrolo|2,Za|Ipyrimidin-4(3ZH)-one. "H -NMR (200 MHz, DMSO-iv) 5: 2.17 (s, m

ЗН), 2.21 (с, ЗН), 7.31 (m, 2Н), 8.12 (m, 2Н), 11.47 (с, 1Н); MS(EN): 258.2 (M 1). 5,6-Dimethyl-2-(3-fluorophenyl)-7H-pyrrolo|2,Za|pyrimidin-4(ZH)-one. "H-NMR (200 MHz, DMSO-dv) 5: 2.18 (s, s

ZN), 2.21 (s, ZN), 7.33 (m, 1H.), 7.52 (m, 1H), 7.85-7.95 (m, 2H), 11.56 (s, 1H), 11.80 (c, 13); MS(EN): 258.1 (M 1). - 5,6- Dimethyl-2-(2-fluorophenyl)-7H-pyrrolo(|2,Za|pyrimidin-4(ZH)-one. "H-NMR (200 MHz, DMSO-dv) 5: 2.18 (with,

ЗН), 2.22 (s, ЗН), 7.27-7.37 (m, 2Н), 7.53 (m, 1Н), 7.68 (m, 1Н), 11.54 (s, 1Н ), 11.78 (c, 1H); MS(EN): 258.1 (M 1). 5,6-Dimethyl-2-isopropyl-7H-pyrrolo|2, ZaIpyrimidin-4(ZH)-one. "H-NMR (200 MHz, DMSO-ydv) 5: 1.17 (d, U-6.6 « 20 Hz, 6H), 2.11 (s, ZN), 2.15 (s, ZN), 2.81 (m, 1H), 11.20 (s, 1H), 11.39 (s, 1H); MS(EN): 206.1 (M 1). w-in s 5,6- Dimethyl- 7H-pyrrolo|2,Zapyrimidine-4(ZN)-one.. "H-NMR (200 MHz, DMSO-yv) 85: 2.13 (s, ZN), 2.17 (s, ZN), c 7 .65 (c, 1H); MS(EN): 164.0 (M 1). "» Preparative example 11:

A solution of 5,6-dimethyl-2-phenyl-7H-pyrrolo|(2,34|Ipyrimidin-4(ZH)-one (1.0g, 4.2mmol) in phosphorus oxychloride (25.0ml) was heated under reflux for b hours and then concentrated to dryness in vacuo Water was added to the residue to initiate crystallization and the resulting solid was filtered and collected to give 0.90 g (8396) of 4-chloro-5,6-dimethyl-2-phenyl-7H -pyrrolo|2,Za|Ipyrimidine. "H-NMR (200 MHz, DMSO-aiv) 5: 2.33 (s, ZN), 2.33 (s, ZN), 7.46-7.49 (m . 11: (4) 50 4-Chloro-5-methyl-2-phenyl-7H-pyrrolo|2,Za|pyrimidine MS(EN): 244.0 (M'-1).

F 4-Chloro-b-methyl-2-phenyl-7H-pyrrolo(2,Za) Pyrimidine. MS(EN): 244.0 (M 1). 4-Chloro-2-phenyl-7H-pyrrolo|2,Za| Ipyrimidine. "H-NMR (200 MHz, DMSO-ab) 5: 8.35 (2.2H), 7.63 (shr.s, 1H), 7.45 (m, 3H), 6.47 (shr. .c, 1H); MS(EN): 230.0 (Ma-n1). 4-Chloro-5,6-dimethyl-2-(3-pyridyl)-7H-pyrrolo|(2,Zapyrimidine. MS(EN ): 259.0 (M -1). 4-Chloro-5,6-dimethyl-2-(2-furyl)-7H-pyrrolo|2,Za|Ipyrimidine. "H-NMR (200 MHz, DMSO-ab ) 5: 2.35 (s, 2H), o 2.35 (s, ZN), 6.68 (dc, 9U-1.8, 3.6 Hu, 1H), 7.34 (dd, 9U- 1.8 Hz, 3.6 Hz, 71H), 7.89 (dd, 9U-1.8, 3.6 Hz, 1H); o MS(EN): 248.0 (M'1). 4- Chloro-5,6-dimethyl-2-(3-furyl)-7H-pyrrolo|2,Za|Ipyrimidine. "H NMR (200 MHz, DMSO-ab) δ: 2.31 (s, ЗН), bo 2 .31 (s, ZN), 6.62 (s, 1H), 7.78 (s, 1H), 8.18 (s, 1H), 12.02 (s, 1H); MS(EN): 248 ,1 (M"-1). 4-Chloro-5,6-dimethyl-2-cyclopentyl-7H-pyrrolo|2,Za|pyrimidine. "H-NMR (200 MHz, DMSO-dv) 5: 1.61 -1.96 (m, 8H), 2.27 (s, ЗН), 2.27 (s, ЗН), 3.22 (m, 1Н), 11.97 (s, 13); MS(ЕН) : 250.1 (M -1). 4-Chloro-5,6-dimethyl-2-(2-thienyl)-7H-pyrrolo|2,Za)pyrimidine. "H-NMR (200 MHz, AMSO-yv) 5: 2.29 (s, ЗН), 65 2.31 (s, ЗН), 7.14 (dd, 9-31 Hz, 4.0 Hz, 1Н), 7.33 ( d, 9U-4.9 Hz, 1), 7.82 (d, 93.1 Gu, 1H), 12.19 (s, 1H);

MEN): 264.1 (M"1).

4-Chloro-5,6-dimethyl-2-(3-thienyl,)-7H-pyrrolo (|2,34| pyrimidine. "H-NMR (200 MHz, DMSO-iv) δ: 2.32 (s, 2H), 2.32 (s, ЗН), 7.62 (dd, U-3.0, 5.2 Hz, 1H), 7.75 (d, 9U-5.2 Hz, 1H), 8, 20 (d, 9-350 Hz, 1H); MS(EN): 264.0 (M 1). 4-Chloro-5,6-dimethyl-2-(4-fluorophenyl)-7H-pyrrolo|2,Za |-pyrimidine. "H-NMR (200 MHz, DMSO-iv) 58: 2.33 (s, 9 ЗН), 2.33 (s, ЗН), 7.30 (m, 2Н), 8.34 ( m, 2H), 12.11 (s, 1H); MS(EN): 276.1 (M 1). 4-Chloro-5,6-dimethyl-2-(3-fluorophenyl)-7H-pyrrolo|2 ,Za|-pyrimidine. "H-NMR (200 MHz, DMSO-iv) 585: 2.31 (s,

ЗН), 2.35 (с, ЗН), 7.29 (m, 1Н), 7.52 (m, 1Н), 7.96 (m, 1Н), 8.14 (m, 1Н), 11, 57 (p. 18); MS(EN): 276.1 (M 1). 4-Chloro-5,6-dimethyl-2-(2-fluorophenyl)-7H-pyrrolo|2,3a|-pyrimidine. "H-NMR (200 MHz, DMSO-iv) 5: 2.34 (s, 70 ..2Н), 2.34 (s, ЗН), 7.33 (m, 2Н), 7.44 (m, 1H), 7.99 (m, 1H), 12/23 (s, 1); MS(EN): 276.1 (M" 1). 4-Chloro-5,6-dimethyl-2-isopropyl-7H-pyrrolo|2,Za|pyrimidine. "H-NMR (200 MHz, DMSO-ydv) 5: 1.24 (d, U-6.6 Hz,

EN), 2.28 (s, ЗН), 2.28 (s, ЗН), 3.08 (kv, U-6.6 Hz, 1Н), 11.95 (s, 1Н); MS(EN): 224.0 (M"-1). 4-Chloro-5,6-dimethyl-7H-pyrrolo (|2,34| pyrimidine. "H-NMR (200 MHz, DMSO-div) 5: 2.31 (s, ZN), 2.32 (s, ZN), 8.40 (s, 18), MS(EN): 182.0 (M), α1-4-chloro-5,6-dimethyl -2-phenyl-7H-7-(1-phenylethyl)pyrrolo|2, Z|pyrimidine.

Preparation example 12:

Di-tert-dicarbonate (4 ,80g, 22,O0mmol) at room temperature. The resulting mixture was stirred for 14 hours. Dioxane was removed in vacuo. The precipitate was filtered off and the filtrate was concentrated to dryness under vacuum. The residue was triturated with EtOAc and then filtered. The filtrate was concentrated to dryness to obtain a mixture of /41-1-amino-2-(1,1-dimethylethoxy)carbonylaminopropane (VK a1-2-amino-1-(1,1-dimethylethoxy)carbonylaminopropane), which could not be separated by conventional This mixture was used for the reaction in Example 8. p

Preparation example 13:

A few drops of M, M-dimethylformamide were added to a solution of Ethos-p-AIa-OH (1.0g, 3.212mmol) and oxalyl chloride (0.428g, 0.29ml, 3.37mmol) in dichloromethane (o) (20.00ml) at 09S. The mixture was stirred at room temperature for 1 hour, and then cyclopropylmethylamine (0.229g, 0.28ml, 3.212mmol) and triethylamine (0.65g, 0.90ml, 6.424mmol) were added. After 10 minutes, the mixture was treated with 1M hydrochloride (10.0ml) and the aqueous mixture was extracted with dichloromethane (3x30.0ml). The organic solution was concentrated to dryness under vacuum. The residue was treated with a solution of 20905 piperidine in She

M, M-dimethylformamide (20, Oml) for 0.5 g. After removal of the solvent in vacuo, the residue was treated with 1 M HCl (20.00 mL) and ethyl acetate (20.00 mL). The mixture was separated and the aqueous layer was alkalized with solid sodium hydroxide to pH-8. The precipitate was removed by filtration and the aqueous solution was subjected to ion exchange with column chromatography, 2090 pyridine was eluted, resulting in 0.262 g (5775) of amide

M-cyclopropylmethyl-p-alanine. "H-NMR (200 MHz, CO500) 5: 0.22 (m, 2H), 0.49 (m, 2H), 0.96 (m, 2H), 2.40 (t, 2H), 2, 92 (t, 2H), 3.05 (d, 2H), MS(EN): 143.1 (M 1).

Preparative example 14: "

M-tert-butoxycarbonyl,-trans-1,4-cyclohexyldiamine.

Trans-1,4-cyclohexyldiamine (6.08 g, 53.2 mmol) was dissolved in dichloromethane (10 mL). Then, a solution of di-tert-butyl dicarbonate (2.32 g, 10.65 mmol in 40 ml of dichloromethane) was added using a cannula. After 20 hours, the "" reaction mixture was partitioned between SNI" with water. The layers were separated and the aqueous layer was extracted with SNI" with (three times). " The combined organic layers were dried with Mo950, filtered and concentrated to give 1.20 g of solid matter (5395). "H-NMR (200 MHz, SOSIv) 6:1.0-1.3 (m, 4H), 1.44 (s, 9H), 1.8-2.1 (m, 4H), 2.62 (screen, 75 1H), 3.40 (width, 1H), 4.37 (width, 1H); MS(EN): 215.2 (M"--1). - 4-(M-acetyl)-M-tert-butoxycarbonyl,-trans-1,4-cyclohexyldiamine. co M-tert-Butoxycarbonyl,-trans-1,4-cyclohexyldiamine (53mg, 2.47mmol) was dissolved in dichloromethane (20ml).

Then acetic anhydride (25 Ω, 2.60 mmol) was added dropwise. After 16 hours, the reaction mixture was diluted - with water and SNCI3. The layers were separated and the aqueous layer was extracted with CHO" (three times). The combined organic layers were dried (4) 50 Ma5O, filtered and concentrated. After recrystallization (EUN/H 20), 190 mg of white crystals were obtained

F (3095). "H-NMR (200 MHz, SOSIv) 5: 0.9-1.30 (m, 4H), 1.43 (s, 9H), 1.96-2.10 (m, 7H), 3.40 (width s, 1H), 3.70 (width s, 1H), 4.40 (width s, 1H), 4.40 (width s, 18); MS(EN): 257.2 ( M"1), 242.1 (M'-15), 201.1 (M'-56). 4-(4-trans-acetamidocyclohexyl)amino-5,6-dimethyl-2-phenyl-7H-(1-phenylethyl)pyrrolo|2, Z|pyrimidine. 4-(M-acetyl)-M-tert-butoxycarbonyl,-trans-1,4-cyclohexyl-diamine (19mg, 0.74mmol) was dissolved in dichloromethane (5ml) and diluted with TEA (bml). After 16 hours, the reaction mixture was concentrated. Raw solid

F) substance, DMSO (2 ml), ManSO 3 (200 mg, 2.2 mmol) and 4-chloro-5,6-dimethyl-2-phenyl-7H-pyrrolo-(2,34|pyrimidine ka) (35 mg, 0, 14 mmol) were combined in a flask and heated to 1302. After 4.5 hours, the reaction mixture was cooled to room temperature and diluted with EtOAc and water. The layers were separated and the aqueous layer was extracted with EtOAc (three times). The combined organic layers dried Mo95O;, filtered and concentrated. After chromatography (preparative silica plates, 20:1, SNCI3:EN), 0.Zmg of a brownish solid was obtained (yield 19605).

MS(EN): 378.2 (M'1). 4-(M-methanesulfonyl)-M-tert-butoxycarbonyl,-trans-1,4-cyclohexyldiamine.

Trans-1,4-cyclohexyldiamine (530 mg, 2.47 mmol) was dissolved in dichloromethane (200 mL) and diluted with pyridine 65 (23 µg, 3 mmol). Then methanesulfonyl chloride (300 mg, 2.60 mmol) was added dropwise. After 16 hours, the reaction mixture was diluted with water and SNCI 3. The layers were separated and the aqueous layer was extracted with SNCI»z (three times).

The combined organic layers were dried over MoSO, filtered and concentrated. After recrystallization (EUN/H 20), 20 mg of white crystals (2995) were obtained. "H-NMR (200 MHz, SOSI) 5: 1.10-1.40 (m, 4H), 1.45 (s, 9H), 2.00-2.20 (m, 4H), 2.98 (s, ZN), 3.20-3.50 (shr.s, 2H), 4.37 (shr.s, 1H); MS(EN): 293.1 (Mk), 278.1 (M' -15). 4-(M-sulfonyl)-M-tert-butoxycarbonyl,-trans-1,4-cyclohexyldiamine (20 mg, 0.7 mmol) was dissolved in dichloromethane (bml) and diluted with TEA (bml). After 16 hours, the reaction mixture was concentrated . The crude reaction mixture, e:c) (2ml), Mmanso»z (10Omg, 1.Tmmol) and 70. 1-chloro-5,6-dimethyl-2-phenyl-7H-pyrrolo-(2,Za|pyrimidine were combined in a flask and heated to 1302C. After 15 hours, the reaction mixture was cooled to room temperature and diluted with E(OAc (three times). The combined organic layers were dried with MazoO), filtered and concentrated. After chromatography (preparative plates with silicon dioxide, 20:1, SNSIZ/EUN) yielded 2.bmg of a brownish solid (yield 595). MS(EN): 414.2 (M"-1)

Example 1: a solution of 4-chloro-5,6-dimethyl-2-phenyl-7H-pyrrolo|2,Za|pyrimidine (0.5Og, 1.94mmol) and 4-trans-hydroxycyclohexylamine (2.23g, 194mmol ) in methyl sulfoxide (10.Oml) was heated at 13023 for 5 hours. After cooling to room temperature, water (10.0 ml) was added and the resulting aqueous solution was extracted with EtOAc (3 x 10.0 ml). The combined solution of EFOAc was dried with M4oaBzO,); and filtered, the filtrate was concentrated to dryness in vacuo, and the residue was chromatographed on silica gel to give 0.49 g (75905) 4-(4-trans-hydroxycyclohexyl)amino-5,b-dimethyl-2-phenyl-7H-pyrrolo (2.341 pyrimidine , mp 197-1999C; "H-NMR (200 MHz, SOSI5) 5: 1.25-1.59 (m, 8H), 2.08 (s, ЗН), 2.29 (s, ЗН ), 3.68-3.79 (m, 1H), 4.32-4.38 (m, 1), 4.88 (d, 9-8 Hz, 1H), 7.26-7.49 ( m, ZN), 8.40-8.44 (dd, U-2.2, 8 Hz, 2H), 10.60 (s, 1H), MS(EN): 337.2 (M"-1) .

The following compounds were obtained by a method similar to that described in example 1: c 4-(4-trans-hydroxycyclohexyl)amino-b6-methyl-2-phenyl-7H-pyrrolo-(2,Za|pyrimidine. TN-NMR (200 MHz, ( 5)

SOSIV) 8: 11.97 (s, 1H, pyrrole-MH), 8.45 (m, 2H, Ag-H), 7.55 (m, 3H, AgH), 6.17 (s, 1H, pyrrole -H), 4.90 (lat, 1H, MH), 4.18 (m, 1H, CH-O), 3.69 (m, 1H, CH-M), 2.40-2, 20 (m, 2H), 2.19-1.98 (m, 2H), 2.25 (s, ZN, SNU), 1.68-1.20 (m, 4H); MS(EN): 323.2 (M'1). zo 4-(4-trans-hydroxycyclohexyl)amino-5-methyl-2-phenyl-7H-pyrrolo-(2,34|pyrimidine. "H-NMR (200 MHz,

SOSI.) 8: 11.37 (s, 1H, pyrrole-MH), 8.40 (m, 2H, Ag-H), 7.45 (m, ЗН, Ag-H), 5.96 (s, 1H, pyrrole-H), 490 "so (lat, 1H, MH), 4.18 (m, 1H, CH-O), 3.69 (m, 1H, CH-M), 2.38 -2.20 (m, 2H), 2.18-1.98 (m, 2H), 2.00 (m, ZN, SNU), and 1.68-1.20 (m, 4H); MS(EN): 323.2 (M"-1). 4-(4-trans-hydroxycyclohexyl)amino-2-phenyl-7H-pyrrolo|2, Za|pyrimidine, m.p. 245.5-245, 6eC; s "H-NMR (200 MHz, SOSIv) 85: 8.33 (m, 2H, Ag-H), 7.42 (m, ZN, Ag-H), 7.02 (d, 1H, 9U -3.6 Hz, pyrrole-H), 6.53 Her (d, 1H, (3.6 Hz, pyrrole-H), 4.26 (m, 1H, CH-O), 3.62 (m, 1H, CH-M), 2.30-2.12 (m, 2H), 2.12-1.96 (m, 2H), 1.64-1.34 (m, 4H); MS, M. -1-309.3; analysis (Si8NooM,O), C, H, M. 4-(4-trans-hydroxycyclohexyl)amino-5, b-dimethyl-2-(3-pyridyl-7H-pyrrolo|2, For) pyrimidine.

TN-NMR (200 MHz, SOSIv) 5: 1.21-1.54 (m, 8H), 2.28 (s, ЗН), 2.33 (s, ЗН), 3.70 (m, 1Н) , 4.31 (m, 1H), 70 4.89 (d, 1H), 7.40 (m, 1H), 8.61 (m, 2H), 9.64 (m, 1H); MS(EN): 338.2 (M 1). n- c 4-(d-trans-hydroxycyclohexyl)amino-5,6-dimethyl-2-(2-furyl)-7H-pyrrolo|2,34| pyrimidine "H-NMR (200 MGu, :z" SOSIv) 6: 1.26-1.64 (m, 8H), 2.22 (s, ZN), 2.30 (s, ZN), 3.72 ( m, 1H), 4.23 (m, 1H), 4.85 (d, 1H), 6.52 (m, 1H), 7.12 (m, 1H), 7.53 (m, 1H), 9.28 (s, 1H); MS(EN): 327.2 (M --1). 4-(4-trans-hydroxycyclohexyl)amino-5,6-dimethyl-2-(3-furyl)-7H -pyrrolo|2.34| pyrimidine. "H-NMR (200 MGu, -I SOSIv) 8: 1.25-1.63 (m, 8H), 2.1.1 (s, ЗН), 2.27 (s , ЗН), 3.71 (m, 71Н), 4.20 (m, 1Н), 4.84 (d, 1Н), 7.03 (m, 7 1Н), 7.45 (m, 1Н), 8.13 (m, 1H), 10.38 (m, 1H); MS(EN): 327.2 (M"1). 4-(4-trans-hydroxycyclohexyl)amino-5,b-dimethyl-2-cyclopentyl-7H-pyrrolo|2,Za|pyrimidine. - "H-NMR (200 MHz, SOSIV) 5: 1.26-2.04 (m, 16H), 2.26 (s, ЗН), 2.27 (s, ЗН), 3.15 (m, 1Н), 3, 70 (m, 1H), 9) 50 4.12 (m, 1H), 4.75 (d, 1H); MS(EN): 329.2 (M 1).

F 4-(d-trans-hydroxycyclohexyl)amino-5,b-dimethyl-2-(2-thienyl)-7H-pyrrolo|2,Za|pyrimidin-4-amine. "H-NMR (200

MHz, SOSI5) 5: 1.28-1.59 (m, 8H), 2.19 (s, ЗН), 2.29 (s, ЗН), 3.74 (m, 1Н), 4.19 ( m, 1H), 4.84 (d, 1), 7.09 (m, 1H), 7.34 (m, 1H), 7.85 (m, 1H), 9.02 (s, 1H); MS(EN): 343.2 (M"-1). 5B 4-(4-trans-hydroxycyclohexyl)amino-5,b-dimethyl-2-(3-thienyl)-7H-pyrrolo|2,Za| Ipyrimidine. T1H-NMR (200

MHz, SOSI5) 5: 1.21-1.60 (m, 8H), 1.98 (s, ЗН), 2.23 (s, ЗН), 3.66 (m, 1Н), 4.22 ( m, 1H), 7.27 (m, 1H), 7.86

IF) (m, 1H), 8.09 (m, 1H), 11.23 (s, 1H); MS(EN): 343.2 (M'-1). ko 4-(4-trans-hydroxycyclohexyl) amino-5, b-dimethyl-2-(4-fluorophenyl)-7H-pyrrolo|2,Za|Ipyrimidine. TN-NMR. (200

MHz, SOSI5) 5: 1.26-1.66 (m, 8H), 1.94 (s, ЗН), 2.28 (s, ЗН), 3.73 (m, 1Н), 4.33 ( m, 1H), 4.92 (d, 1H), 7.13 60 (m, 2H), 8.41 (m, 2.H), 11.14 (s, 1H); MS(EN): 355.2 (M"1). 4-(4-trans-hydroxycyclohexyl)amino-5,b-dimethyl-2-(3-fluorophenyl)-7H-pyrrolo(|2,Za| Ipyrimidine. "H-NMR (200

MHz, SOSIv) 5: 1.26-1.71 (m, 8H), 2.06 (s, ЗН), 2.30 (s, ЗН), 3.72 (m, 1Н), 4.30 ( m, 1H), 4.90 (d, 1), 7.09 (m, 1), 7.39 (m, 1H), 8.05 (m, 1H), 8.20 (m, 1H), 10.04 (p. 18); MS(EN): 355.2 (M 1). b5 4-(4-trans-hydroxycyclohexyl)amino-5, 6b-dimethyl-2-(2-fluorophenyl)-7H-pyrrolo|2,34)pyrimidine. "H-NMR (200

MHz, SOSI5) 5: 1.30-1.64 (m, 8H), 2.17 (s, ЗН), 2.31 (s, ЗН), 3.73 (m, 1Н), 4.24 ( m, 1H), 4.82 (d, 1H), 7.28

(m, 2H), 8.18 (m, 1H), 9.02 (m, 1H), 12.20 (s, 1H); MS(EN): 355.3 (M 1). 4-(d-trans-hydroxycyclohexyl)amino-5,6-dimethyl-2-isopropyl-7H-pyrrolo|2,Za|Ipyrimidine. H-NMR (200 MGu,

SOSIv) 8: 1.931 (d, 9U-7.0 Gu, 6H), 1.30-165 (m, 8H), 2.27 (s, ЗН), 2.28 (s, ЗН), 3.01 (m, 9U-7.0 Gu, 1), 53.71 (m, 1H), 4.14 (m, 1H), 4.78 (d, 1H); MS(EN): 303.2. a1-4-(2-trans-hydroxycyclohexyl)amino-5,6-dimethyl-2-isopropyl-7H-pyrrolo|2,Za|Ipyrimidine. H-NMR (200

MHz, SOSI5) 5: 1.31-1.42 (lat., 4H), 1.75-1.82 (lat., 4H), 2.02 (s, ZN), 2.29 (s, ZN) , 3.53 (m, 1H), 4.02 (m, 1H), 5.08 (d, 1H), 7.41-7 48(m, ЗН), 8.30 (m, 4H), 10 .08 (c, 1H); MS(EN): 337.2 (M1). / malu yt proxicyclo vky lian no podimetil tet tri pyrimidine. MS(EN) 3532 4-(3,4-cis-dihydroxycyclohexyl)amino-5,6-dimethyl-2-phenyl-7H-pyrrolo|2,34| pyrimidine MOS(EN): 35332 (M). 4-(2-acetylaminoethyl)amino-5,6-dimethyl-2-phenyl-7H-pyrrolo|2,3a|-pyrimidine, m.p. 196-19996;

TN-NMR (200 MHz, SOSIv) 5: 1.72 (s, НН), 1.97 (s, НН), 2.31 (s, НН), 3.59 (m, 2Н), 3.96 (m, 2H), 5.63 (width, 1H), 7.44-7.47 (m, ЗН), 8.36-8.43 (dd, 9-1 Hz, 7 Hz, 2H), 10.76 (s, 1H); MS(EN): 324.5 (M"1). a1-4-(2-trans-hydroxycyclopentyl)amino-5, b-dimethyl-2-phenyl-7H-pyrrolo|2,3a4| pyrimidine!". "H-NMR (200 MHz,

SOSIia) 5: 1.62 (m, 2H), 1.79 (width, 4H), 1.92 (s, ЗН), 2.29 (s, ЗН), 4.11 (m, 1Н), 4 .23 (m, 1H), 528 (d, 1H), 7.41-7.49 (m, ЗН), 8.22 (m, 2H), 10.51 (s, 1H); MS(EN): 323.2 (M'1). 1 For the preparation of 2-trans-hydroxycyclopentylamine, see PCT 9417090. a1-4-(3-trans-hydroxycyclopentyl)amino-5,6-dimethyl-2-phenyl-7H-pyrrolo|2,3a| pyrimidine'. "H-NMR (200 MHz,

SOSIv) 5: 1.58-1.90 (width, 6H), 2.05 (s, ZN), 2.29 (s, ZN), 4.48-4.57 (m, 1H), 4, 91-5.01 (m, 2H), 7.35-7.46 (m, ЗН), 8.42-8.47 (m, 2H), 10.11 (c, 1H); MS(EN): 323.2 (M"1). c 1 For the preparation of 2-trans-hydroxycyclopentalamine, see EP-A-322242. o a1-4-(3-cis-hydroxycyclopentyl)amino-5,6-dimethyl -2-phenyl-7H-pyrrolo|2,Za|pyrimidine 7. IN-NMR (200. MHz,

SOSIv) 5: 1.82-2.28 (width, 6H), 2.02 (s, ZN), 2.30 (s, ZN), 4.53-4.60 (m, 1H), 4, 95-5.08 (m, 1H), 5.85-5.93 (d, 1H), 7.35-7.47 (m, ЗН), 8.42-8.46 (m, 2H), 10.05 (c, 1H); MS(EN): 323.2 (M --1). 1 For the preparation of 3-cis-hydroxycyclopentylamine, see ER-A-322242. ish 4-(3,4-trans-dihydroxycyclopentyl)amino-5,6-dimethyl-2-phenyl-7H-pyrrolo|2,3a| pyrimidine'. "H-NMR (200 MHz, "o

SOSIv) 8:1.92-1.99 (width, 2H), 2.14 (s, ZN), 2.20 (width, 2H), 2.30 (s, ZN), 2.41-2, 52 (width, 2H), 4.35 (m, 2H), m 4.98 (m, 2H), 7.38-7.47 (m, ZH), 8.38-8.42 (m, 2H ), 9.53 (m, 1H); MS(EN.): 339.2 (M --1). 1 For the preparation of 3,4-trans-dihydroxycyclopentylamine, see PCT 9417090. c 4-(3-amino-3-oxopropyl)amino-5,6-dimethyl-2-phenyl-7H-pyrrolo-|(2,34| pyrimidine. "H-NMR (200 MHz, SOCIi) 5 : che 2.02 (s, ZN), 2.29 (s, ZN), 2.71 (t, 2H), 4.18 (m, 2H), 5.75-5.95 (m, ZN) , 7.38-7.48 (m, ZN), 8.37-8.41 (m, 2H), 10.42 (s, 1H); MS(EN): 310.1 (M). 4- (3-M-Cyclopropylmethylamino-3-oxopropyl)amino-5, b-dimethyl-2-phenyl-7H-pyrrolo (2.341) pyrimidine". "H-NMR " (200 MHz, CO300) 5: 0.51 (sq , 2H), 0.40 (sq., 2H), 1.79-1.95 (width, 1H), 2.36 (s, ZH), 2.40 (s, ZH), 2.72 (t, 2H), 2.99 (d, 2H), 4.04 (t, 2H), 7.58-7.62 (m, ЗН), 8.22-8.29 (m, 2H); MS(EN): 364.2 (M 1). c 4-(2-amino-2-oxoethyl)amino-5, b-dimethyl-2-phenyl-7H-pyrrolo (2.34) pyrimidine. "H- NMR (200 MHz, SOzOB) 8: ;» 2.31 (s, ЗН), 2.38 (s, ЗН), 4.26 (s, 2Н), 7.36 (m, ЗН), 8.33 (m, 2H); MS(EN): 396.1 (M 1). 4-(2-M-methylamino-2-oxoethyl)amino-5,6-dimethyl-2-phenyl-7H-pyrrolo|2, 34|pyrimidine.'H-NMR (200 MGu,

SOS) 8: 1.99 (s, ZN), 2.17 (s, ZN), 2.82 (d, ZN), 4.39 (d, 2H), 5.76 (t, 1H), 6 .71 (width, 1H), 7.41-7.48 - (m, ЗН), 8.40 (m, 2H), 10.66 (s, 1H); MS(EN): 310.1 (M"1). ka 4-(3-tert-butyloxyl-3-oxopropyl)amino-5,6-dimethyl-2-phenyl-7H-pyrrolo|2,Za|Ipyrimidine. "H-NMR (200 MGu,

SOSIv) 6: 1.45 (s, 9H), 1.96 (s, ЗН), 2.29 (s, ЗН), 2.71 (t, 2Н), 4.01 (q, 2Н), 5 .78 (t, 1H), 7.41-7.48 (m,

In ZN), 8.22-8.29 (m, 2N.); MS(EN): 367.2 (M'1). o 20 4-(2-hydroxyethyl)amino-5,6-dimethyl-2-phenyl-7H-pyrrolo|2, ZaIpyrimidine. "H-NMR (200 MHz, SOSIv) 8: 1.92 (s,

Ф ЗН), 2.29 (s, ЗН), 3.81-3.98 (width, 4Н), 5.59 (t, 1Н), 7.39-7.48 (m, ЗН), 8 .37 (m, 2H), 10.72 (s, 18); MS(EN): 283.1 (M). 4-(3-Hydroxypropyl)amino-5,6-dimethyl-2-phenyl-7H-pyrrolo|2,Za|I-pyrimidine. 8 "H-NMR (200 MHz, SOCIv) 8: 1.84 (m, 2H), 1.99 (s, ЗН), 2.32 (s, ЗН), 3.62 (t, 2Н), 3.96 (m, 2Н), 3.35 (t, 1Н), 7, 39-7.48 (m, ZN), 8.36 (m, 2. o H), 10.27 (s, 1H); MS(EN): 297.2 (M'1). 4-(4 -hydroxybutyl)amino-5,6-dimethyl-2-phenyl-7H-pyrrolo|2,Za|pyrimidine. "H-NMR (200 MHz, SOCIv) 8: 1.71-1.82 (m, 4H) , 1.99 (s, ЗН), 2.31 (s, ЗН), 3.68-3.80 (m, 4Н), 5.20 (t, 1), 7.41-7.49 (m , ZN), 8.41 (m, in 2H), 10.37 (s, 1H); MS(EN): 311.2 (M"1). 4-(4-t-trans-acetylaminocyclohexyl)amino-5,6-dimethyl-2-phenyl-7H-pyrrolo|2, Z|pyrimidine. 4-( 4-trans-methylsulfonylaminocyclohexyl)amino-5,6-dimethyl-2-phenyl-7H-pyrrolo|2,Za|pyrimidine 4-(2-acetylaminoethyl)amino-5,6-dimethyl-2-phenyl-7H -7-(1-phenylethyl)pyrrolo|2,34|pyrimidine 4-(4-trans-hydroxycyclohexyl)amino-5,6-dimethyl-2-phenyl-7H-1-(phenylethyl)pyrrolo(|2, Za |pyrimidine. 4-(3-pyridylmethyl)amino-5,6-dimethyl-2-phenyl-7H-7-(1-phenylethyl)-pyrrolo|2, Za|Ipyrimidine. 65 4-(2-methylpropyl)amino -5,6-dimethyl-2-phenyl-7H-7-(1-phenylethyl)-pyrrolo|2,Za|pyrimidine.

Example 2

To a mixed suspension of triphenylphosphine (0.047g, 0.179mmol) and benzoic acid (0.022g, 0.179mmol) in

4-(4-trans-hydroxycyclohexyl)amino-5,6-dimethyl-2-phenyl-7H-pyrrolo|2,Za|pyrimidine (0.05g, 0.149mmol) was added to THgF (1.0ml) cooled to 0 at 02. Then diethyl azodicarboxylate (0.028 ml, 0.179 mmol) was added dropwise over 10 minutes. The reaction mixture was left to warm to room temperature. After completion of the reaction, which was indicated by TLC, the reaction mixture was quenched with an aqueous solution of sodium bicarbonate (33.0 ml). The aqueous phase was separated and extracted with ether (2 x 5 ml). The organic extracts were combined, dried and concentrated in vacuo to dryness. Ether (2.Oml) and hexane (5.Oml) were added to residue 70, after which the excess triphenylphosphine oxide was filtered off. After concentration of the filtrate, a viscous oil was obtained, which was purified by column chromatography (hexane, cetyl acetate E 41) and 5.0 mg (7.690) of 4-(4-cis-benzoyloxycyclohexyl)amino-5,6-dimethyl-2-phenyl was obtained -7H-pyrrolo (2.34| pyrimidine. MS(EN): 441.3 (M).

As a result of the reaction, 50.0 mg of (8496) 75 4-(3-cyclohexyl)amino-5,6-dimethyl-2-phenyl-7H-pyrrolo|2,Za|pyrimidine was also obtained. MS(EN): 319.2 (M).

Example C

To a solution of 4-(4-cis-benzoyloxycyclohexyl)amino-5,6-dimethyl-2-phenyl-7H-pyrrolo|2,3a|Ipyrimidine (5.Omg, 0.0114 mmol) in ethanol (1.0 ml) was added 10 drops of 2M sodium hydroxide. After 1 hour, the reaction mixture was extracted with ethyl acetate (3 x 5.0ml) and the organic layer was dried, filtered and concentrated in vacuo to dryness. The residue was subjected to column chromatography (hexane-ethyl acetate - 4:1) to obtain 3,bmg (94965) 4-(4-cis-hydroxycyclohexyl)amino-5,6-dimethyl-2-phenyl-7H-pyrrolo|2,3a| -pyrimidine. MS(EN): 337.2 (M-1).

The following compounds were obtained by the method described in Example 3. 4-(3-M,M-Dimethyl-3-oxopropyl)amino-5,6-dimethyl-2-phenyl-7H-pyrrolo (|2,34| pyrimidine. "H -NMR (200 MGu, sc

SOSIv) 5: 2.01 (s, ЗН), 2.31 (s, ЗН), 2.73 (t, 2Н), 2.97 (s, 6Н), 4.08 (m, 2Н), 6 .09 (t, 1H), 7.41-7.48 (m,

ЗН), 8.43 (m, 2Н), 10.46 (s, 1Н); MS(EN): 338.2 (M 1). o 4-(2-formylaminoethyl)amino-5,6-dimethyl-2-phenyl-7H-pyrrolo|2,Za|-pyrimidine. H-NMR (200 MHz, SOSIv) 58: 2.26 (s, HN), 2.37 (s, HN), 3.59-3.78 (m, 2H), 3.88-4.01 (m, 2H), 5.48-5.60 (m, 1H), 7.38-7.57 (m, ЗН), 8.09 (s, 1H), 8.30-8.45 (m , 2H), 8.82 (c, 1H); MS(EN): 310.1 (M"-1). ise) 4-(3-acetylaminopropyl)amino-5,6-dimethyl-2-phenyl-7H-pyrrolo-(2,34| pyrimidine; MS(EN) ): 338.2 (M-1). p

Example 4 4-(3-tert-Butyloxy-3-oxopropyl)amino-5,6-dimethyl-2-phenyl-7H-pyrrolo|2/za|Ipyrimidine (70.Omg, 0.191 mmol) - was dissolved in a mixture of trifluoroacetic acid :dichloromethane (1:11, 5.Oml). The resulting solution was stirred at room temperature for 1 hour and then heated under reflux for 2 hours. After cooling to room temperature, the mixture was concentrated to dryness in vacuo. The residue was subjected to - preparative thin-layer chromatography (E(Fashexane"AcOH -7:2.5:0.5) with the receipt of 40.Omg (68965) 4-(3-hydroxy-3-oxopropyl)amino-5,6-dimethyl- 2-phenyl-7H-pyrrolo|2,Za|pyrimidine. "H-NMR (200 MHz, SOZOB) 5: 2.32 (s, ЗН), 2.38 (s, ЗН), 2.81 (t, 2Н) , 4.01 (t, 2H), 7.55 (m, 3H), 8.24 (m, 2H); MS(EN): 311.1 (M). 4: -o c 4-(3-aminopropyl)amino-5,6-dimethyl-2-phenyl-7H-pyrrolo|2,Za|Ipyrimidine. MS(EN): 296.1 (M7--1), 279 ,1 (M'-MNU").

Example 5; t 4-(3-Hydroxy-3-oxophenyl)amino-5,6-dimethyl-2-phenyl-7H-pyrrolo|2,3a|pyrimidine (50.Omg, 0O.161 mmol) was dissolved in a mixture of M, M-dimethylformamide (0.5Oml), dioxane (0.5O0ml) and water (0.25ml). Methylamine (0.02ml, 4Omas.9o in water, 0.242mmol), triethylamine (0.085ml) and M,M,M,-I M'-tetramethyluronium tetrafluoroborate (61.2mg, 0.20Zmmol) were added to this solution. After stirring at room temperature for 10 minutes, the solution was concentrated and the residue was subjected to preparative thin-layer chromatography (E(Ac) to give di 35.Omg (6796). 4-(3-M-methyl-3-oxopropyl)amino-5,6 -dimethyl-2-phenyl-7H-pyrrolo|2,ZaI-pyrimidine. "H-NMR (200 MHz, -I SOCIv) 86: 1.92 (s, HN), 2.30 (s, HN), 2.65 (t, 2H), 4.08 (t, 2H), 5.90 (t, 1), 6.12 (m, 71H), 7.45 (m, ЗН), syu 50 8.41 (m, 2H), 10.68 (s, 1H); MS(EN): 311.1 (M 1).

The following compounds were obtained by the method described in Example 5: ii; 4-(2-dcyclopropanecarbonylaminoethyl)amino-5,6-dimethyl-2-phenyl-7H-pyrrolo|2, Z|pyrimidine. MS(EN): 350.2 (M). 4-(2-isobutyrylaminoethyl)amino-5,6-dimethyl-2-phenyl-7H-pyrrolo-|(2,34| pyrimidine. MS(EN): 352.2 (M -1). 99 4-(Z -propionylaminopropyl)amino-5,6-dimethyl-2-phenyl-7H-pyrrolo-(2,Za|Ipyrimidine. "H-NMR (200 MHz, SOSIV) 8:

HF) 1.00-1.08 (t, ZN), 1.71-2.03 (m, 4H), 2.08 (s, ZN), 2.37 (s, ZN), 3.26- 3.40 (m, 2H), 3.79-3.96 (m, 2H), g 5.53-5.62 (m, 1H), 6.17-6.33 (m, 1H), 7 .33-7.57 (m, ZN), 8.31-8.39 (m, 2H), 9.69 (s, 18); MS(EN): 352.2 (M 1). 4-(2-methylsulfonylaminoethyl)amino-5,6-dimethyl-2-phenyl-7H-pyrrolo|2,34| pyrimidine "H-NMR (200 MGu, 6o SOCI3) 85: 2.18 (s, ZN), 2.27 (s, ZN), 2.92 (s, ZN), 3.39-3.53 (m, 2H), 3.71-3.88 (m, 2H), 5.31-5.39 (m, 1H), 6.17-6.33 (m, 1H), 7.36-7.43 ( m, ZN), 8.20-8.25 (m, 2H), 9.52 (s, 18); MS(EN): 360.2 (M 1).

Example 6

A mixture of 4-Chloro-5,6-dimethyl-2-phenyl-7H-pyrrolo|2,34|-pyrimidine (0.70 g, 2.72 mmol) and 1,2-diaminoethane (10.0 mL, 1500 mmol) was heated under reflux refrigerator in an inert atmosphere for 6 hours. Excess 65 amine was removed in vacuo, and the residue was successively washed with ether and hexane to obtain 0.75 g (98965) of 4-(2-aminoethyl)amino-5,b-dimethyl-2-phenyl-7H-pyrrolo |2.34| pyrimidine MS(EN): 282.2 (M"-1), 265.1 (M'-MNU").

Example 7

To a solution of 4-(2-aminoethyl)amino-5,6-dimethyl-2-phenyl-7H-pyrrolo|2,34|Ipyrimidine (70O,Omg, 0.249mmol) and triethylamine (50.4mg, 0.498mmol) in dichloromethane (2.0 ml) was added propionyl chloride (25 mg, 0.024 ml, 0.274 mmol) at 09C. After 1 hour, the mixture was concentrated in vacuo and the residue subjected to preparative thin-layer chromatography (Eu:oAcC) Kk) yielding 22.0 mg (2690) of 4-(2-propionylaminoethyl)amino-5,6-dimethyl-2-phenyl-7H-pyrrolo -|(2,34)| pyrimidine MS(EN): 338.2 (M 1).

The following compounds were obtained by the method described in Example 7: 4-(2-M'-Methylcarbamidoethyl)amino-5,b-dimethyl-2-phenyl-7H-pyrrolo-|2,34| pyrimidine "H-NMR (200 MGu, 70 SOS) 5:213 (s, ЗН), 2.32 (s, ЗН), 3.53 (d, ЗН), 3.55 (m, 2Н), 3.88 (m, 2H), 4.29 (m, 1H), 5.68 (t, 1H), 5.84 (m, 1H), 7.42 (m, ЗН), 8.36 (dd, 2H) , 9.52 (s, 1H), MS(EN): 339.3 (M 1). 4-(2-M'-ethylcarbamidoethyl)amino-5,6-dimethyl-2-phenyl-7H-pyrrolo|2 .34| pyrimidine. MS(EN): 353.2 (M 1).

Example 8

To a solution of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (41.1 mg, 0.215 mmol), dimethylaminopyridine (2.4 mg, 0.02000 mmol) and pyruvic acid (18.9 mg, 0.015 ml, 0.215 mmol) in dichloromethane ( 2.00 ml) was added 4-(2-aminoethyl)amino-5,6-dimethyl-2-phenyl-7H-pyrrolo(2,Za|Ipyrimidine (55.0mg, 0.196bmmol). The mixture was stirred at room temperature for 4 hours After standard processing and column chromatography (EU:OAcC), 10.00 mg (1596) of 4-(2'-pyruvilamidoethyl)amino-5,6-dimethyl-2-phenyl-7H-pyrrolo(|(2,Za|Ipyrimidine) was obtained MS(EN): 352.2 (M"1).

Example 9

M-trimethylsilyl isocyanate ( 43.3 mg, O0.051 ml, 0.320 mmol). The mixture was stirred at room temperature for 3 hours, and then an aqueous solution of sodium bicarbonate was added. After filtration through a small amount of silica gel, the filtrate was concentrated in vacuo to dryness to obtain 9.8 mg (1496) of 4-(2-carbamidoethyl)amino-5,6-dimethyl-2-phenyl-7H-pyrrolo|2,Za|pyrimidine . MS(EN): 325.2 (M 1). at

The following compounds were obtained by the method described in Example 9: a1-4-2-acetylaminopropyl)amino-5,6-dimethyl-2-phenyl-7H-pyrrolo-(2,Za|pyrimidine. TN-NMR (200 MHz,

SOSIV) 8: 1.28-1.32 (d, 9-8 Gu, ZN), 1.66 (s, ZN), 1.96 (s, ZN), 2.30 (s, ZN), 3 .76-3.83 (m, 2H), 4.10-4.30 "OO (m, 1Hn), 5.60-5.66 (t, 9-6 Hz, 1H), 7.40-7 .51 (m, ZN), 8.36-8.43 (m, 2H), 10.83 (s, 1H); MS(EN): 338.2 (M"1). so (2)-4-(2-acetylaminopropyl)amino-5,6-dimethyl-2-phenyl-7H-pyrrolo-|2,34| pyrimidine.' H-NMR (200 MGu,

SOSIV) 8: 1.91 (d, ZN), 1.66 (s, ZN), 1.99 (s, ZN), 2.31 (s, ZN), 3.78-3.83 (m, 2H), 4.17-4.22 (m, 1H), 5.67 - (t, 1H), 7.38-7.5 (m, ЗН), 8.39 (m, 2H), 10, 81 (c, 1H); MS(EN): 338.2 (M 1). c (2)-4-(1-methyl-2-acetylaminoethyl)amino-5,6-dimethyl-2-phenyl-7H-pyrrolo|2,Za|pyrimidine. H-NMR (200 MGu, ch

SOSIv) 6: 1.41 (d, ZN), 1.68 (s, ZN), 2.21 (s, ZN), 2.34 (s, ZN), 3.46-3.52 (width m, 2H), 4.73 (m, 1), 5.22 (d, 1H), 7.41-7.46 (m, ЗН), 8.36-8.40 (m, 2H), 8 .93 (p. 13); MS(EN): 338.2 (M 1). (8)-4-(2-acetylaminopropyl)amino-5,6-dimethyl-2-phenyl-7H-pyrrolo-|2,34| pyrimidine H-NMR (200 MGu,

SOSIV) 8: 1.91 (d, ZN), 1.66 (s, ZN), 2.26 (s, ZN), 2.35 (s, ZN), 3.78-3.83 (m, 2H), 4.17-4.22 (m, 1H), 5.67 t0 (t, 1H), 7.38-7.5 (m, ЗН), 8.39 (m, 2H), 8, 67 (c, 1H); MS(EN): 338.2 (M"1). 8 s (8)-4-(1-methyl-2-acetylaminoethyl)amino-5,6-dimethyl-2-phenyl-7H-pyrrolo|2,Za |Ipyrimidine. "H-NMR (200 MGu, :z" SOSIZ) 65: 1.41 (d, ZN), 1.68 (s, ZN), 2.05 (s, ZN), 2.32 (s , ЗН), 3.46-3.52 (m, 2Н), 4.73 (m, 1), 5.22 (d, 1Н), 7.41-7.46 (m, ЗН), 8, 36-8.40 (m, 2H), 10.13 (s, 1H); MS(EN): 338.2 (M -1).

Example 10 - I Reaction of 4-chloro-5,6-dimethyl-2-phenyl-7H-pyrrolo|2,Za|pyrimidine with a mixture of a1-1-amino-2-(1,1-dimethoxyethoxy)carbonylaminopropane and where a1- 2-amino-1-(1,1-dimethoxyethoxy)carbonylaminopropane was carried out by the method described in Example 1. -I As a result of this reaction, a mixture of a1-4-(1-methyl-2-(1,1-dimethylethoxy)carbonylamino)ethylamino was obtained -5,6-dimethyl-2-phenyl-7H-pyrrolo|2,3a|pyrimidine and and a1-4-(-2-methyl-2-(1,1-dimethylethoxy)carbonylamino)ethylamino-5,6-dimethyl -2-phenyl-7H-pyrrolo|2,3a|pyrimidine,

F which was separated using column chromatography (E(OAc/hexane-1:3). The first fraction was a1-4--1-methyl-2-(1,1-dimethylethoxy)-carbonylaminoethyl)amino-5,6- dimethyl-2-phenyl-7H-pyrrolo|2,Za)pyrimidine. "H-NMR (200 MHz, SOSIZ) 8: 1.29-1.38 (mM, 12H), 1.95 (s, ZN), 2.31 (s, ZN), 3.34-3.43 (m, 2H), 4.62-4.70 (m, 1H), 5.36-5.40 (d, 1-8 Hz, 1H), 5.53 o (width, 1H), 7, 37-7.49 (m, ZN), 8.37-8.44 (m, 2H), 10.75 (s, 1H), MS(EN): 396.3 (M"--1); The second fraction was a1-4-(2-(1,1-dimethoxyethoxy)carbonylaminopropyl)amino-5,6-dimethyl-2-phenyl-7H-pyrrolo|2,ZaIpyrimidine: "H-NMR o (200 MHz, SOCI5 ) 6: 1.26-1.40 (m, 12H), 2.00 (s, ЗН), 2.31 (s, ЗН), 3.60-3.90 (m, 2Н)3 3.95 -4.10 (m, 1H), 5.41-5.44 (d, 9U-6.0 Hz, 1H), 5.65 (width, 1H), 7.40-7.46 (m, ЗН), 8.37-8.44 (m, 2Н), 10.89 (s, 1Н); MS(ЕН): 396.2 (М 1). bo The following compounds were obtained by the method described in Example 10: ( 8,5)-4-(2-acetylaminocyclohexyl)amino-5,6-dimethyl-2-phenyl-7H-pyrrolo|2,Za|Ipyrimidine. "H-NMR (200 MHz,

SOSIZ) 6: 1.43 (m, 4H), 1.60 (s, ЗН), 1.83 (m, 2Н), 2.18 (s, ЗН), 2.30 (m2Н), 2.32 (c, ЗН), 3.73 (width, 1Н), 4.25 (width, 1Н), 5.29 (d, 1Н), 7.43-7.48 (m, ЗН), 8, 35-8.40 (m, 2H), 9.05 (s, 1H). в5 4-(2-methyl-2-acetylaminopropyl)amino-5,6-dimethyl-2-phenyl-7H-pyrrolo|2,34| pyrimidine "H-NMR (200 MGu,

SOSIv) 5: 1.51 (s, 6H), 1.56 (s, ЗН), 2.07 (s, ЗН), 2.36 (s, ЗН), 3.76 (d, 2Н), 5 .78 (t, 1H), 7.41-7.48 (m,

ЗН), 7.93 (s, 1Н), 8.39 (m, 2Н), 10.07 (s, 1Н); MS(EN): 352.3 (M"-1).

Example 11 a1-4--1-Methyl-2-(1,1-dimethylethoxy)carbonylaminoethyl)amino-5,6-dimethyl-2-phenyl-7H-pyrrolo|2, Za|Ipyrimidine (60.bmg, 0.153mmol ) was treated with trifluoroacetic acid (0.5 ml) in dichloromethane (2.0 ml) for 14 hours.

The organic solvent was removed in vacuo to dryness. The residue was dissolved in M,M-dimethylformamide (2.0ml) and triethylamine (2.0ml). Acetic anhydride (17.2mg, 0.016, 0.169mmol) was added to the solution at 09. The resulting mixture was stirred at room temperature for 48 h, and then concentrated to dryness under vacuum. The residue was subjected to preparative thin-layer chromatography (ELUOAc) to obtain 27.00 mg (52905) 70. a1-4-(1-methyl-2-acetylaminoethyl)amino-5, b-dimethyl-2-phenyl-7H-pyrrolo|2,Za Ipyrimidine. H-NMR (200 MGu,

SOSIv) 6: 1.38-1.42 (d, 9-8 Hz, ZN), 1.69 (s, ZN), 2.01 (s, ZN), 2.32 (s, ZN), 3 ,38-3.60 (m, 2H), 4.65-4.80 (m, 1H), 5.23-5.26 (d, 9-6 Hz, 1H), 7.40-7.51 (m, ZN), 8.37-8.43 (m, 2H) / 10.44 (c, 13); MS(EN): 338.2 (M 1).

Example 12. -chloro-5,6-dimethyl-2-phenyl-7H-pyrrolo|2,Za|pyrimidine (0.15g, 0.58Zmmol) and (18, 21K)-(-)-1,2-diaminocyclohexane (0, 63g, 5.517mmol), was treated with triethylamine (0.726g, 7.175mmol) and acetic anhydride (0.325g, 3.18mmol) in M,M-dimethylformamide (10.00ml) at room temperature for 2 hours.

After removal of the solvent in vacuo, ethyl acetate (10.0 mL) and water (10.0 mL) were added to the residue. The mixture was separated and the aqueous layer was extracted with ethyl acetate (2 x 10.0ml). The combined ethyl acetate solution was dried (Mo5O) and filtered. The filtrate was concentrated to dryness under vacuum and the residue was subjected to column chromatography (EoAc:hexane-1:1) Kk) obtaining 57.0 mg (2690) (Kk,

Kk)-4--2-acetylaminocyclohexyl)amino-5,6-dimethyl-2-phenyl-7H-pyrrolo|2,Za|pyrimidine. TNA-NMR (200 MG,

SOSI)) 5: 1.43 (m, 4H), 1.60 (s, ЗН), 1.84 (m, 2Н), 2.22 (s, ЗН), 2.30 (m, 2Н), 2.93 (s, ЗН), 3.72 (width, рбо1Н), 4.24 (width, 1Н), 5.29 (d, 1Н), 7.43-7.48 (m, ЗН ), 8.35-8.39 (m, 2H), 8.83 (s, 1H); MS(EN): 378.3 (M).

Example 13 o)

Acetic anhydride (0.108 g, 1.0 mmol) at 020.

The resulting mixture was stirred at room temperature for 4 hours and the solvent was removed in vacuo. The residue was subjected to preparative thin-layer chromatography (EuAc':hexane-1:1) to obtain 32.3 mg (7190) of 4-(2-acetyloxyethyl)amino-5,6-dimethyl-2-phenyl-7H-pyrrolo|2 ,For|Ipyrimidine. "H-NMR (200 MHz, SOSIv) 8: 1.90 o (s, ЗН), 2.08 (s, ЗН), 2.31 (s, ЗН), 4.05 (m, 2Н), 4 -

Example 14

A solution of EGtos-p-AIa-OH (97.4 mg, 0.31 μmmol) and oxalyl chloride (39.7 mg, 27.3 μl, 0.31 μmmol) in dichloromethane (4 Oml) with 1 drop of M,M-dimethylformamide was stirred at 09C for 1 hour, and then added - 4-(2-aminoethyl)amino-5, b-dimethyl-2-phenyl-7H-pyrrolo|2,34|Ipyrimidine (80.00 mg, 0.285 mmol) and triethylamine (57.bmg, 79.4 μl, 0.570 mmol) at 02C. After 3 hours, the mixture was concentrated in vacuo and the residue was treated with a solution of 2095 piperidine in M,M-dimethylformamide (2.0 ml) for 0.5 g. After removing the solvent in vacuum, the residue was washed with a mixture of diethyl ether texane (1:5) to obtain Z,Og (3905) 70 4-(6-amino-3-aza-4-oxoethyl)amino-5,6-dimethyl-2- phenyl-7H-pyrrolo-(2,3a|pyrimidine a. MS(EN): 353.2 (M 1). 8 s Example 15

From" A solution of /4-(2-aminoethyl)amino-5,6-dimethyl-2-phenyl-7H-pyrrolo(2,Za|Ipyrimidine / (70.Omg, 0.249 mmol) and succinic anhydride (27.0 mg, 0.274 mmol) in dichloromethane (4.00 ml) with 1 drop of M, M-dimethylformamide was stirred for 4 hours at room temperature. The reaction mixture was extracted with 2095 sodium hydroxide (3x5.00 ml). The aqueous solution was acidified with 3M hydrochloride to pH-7.0. This entire mixture was extracted with ethyl acetate (3x10ml). The combined organic solution was dried (Mo5O) and filtered. The filtrate was concentrated to dryness under vacuum to yield 15.0 mg (16965)-1 4-(7-hydroxy-3-aza) -4,7-dioxoheptyl)amino-5,6-dimethyl-2-phenyl-7/H-pyrrolo |2.34| opirimidine MS(EN) 382.2 (M). (95) 50 Example 16

F To 10 ml of dimethylformamide (DMF) at room temperature was added 7O0 mg of 4-cis-3-hydroxycyclopentyl)amino-2-phenyl-5,6-dimethyl-7H-pyrrolo|2,34|Ipyrimidine, and then added 455 mg

M-Bos-glycine, 20 mg of M,M-dimethylaminopyridine (OMAR), 29 mg of hydroxybenzotriazole (NOVT) and 622 mg of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (EOCI) hydrochloride. The reaction mixture was left overnight with stirring. Then, DMF was removed under reduced pressure and the reaction mixture was partitioned between 20 ml of ethyl acetate and

GF) BOml of water. The aqueous part was extracted with another 2 x 20 ml of ethyl acetate and the combined organic parts were washed

GI with saline solution, dried with anhydrous sodium sulfate, filtered and concentrated. As a result of purification on silica gel and eluting with a mixture of ethyl acetate/hexane, 4100 mg of the desired product was obtained: 4-(cis-3-(M-tert-butoxycarbonyl,-2-aminoacetoxy)cyclopentyl)amino-2-phenyl-5,6-dimethyl-7H -pyrrolo(2,Za|)pyrimidine, MOS(EN): (M'-1) -480.2. Then, the ester was treated with 5 ml of 2095 trifluoroacetic acid in dichloromethane at room temperature, left overnight, after which it was concentrated. After treatment with ethyl acetate, 300 mg of a whitish solid, trifluoroacetic acid salt and 4-(cis-3-(2-aminoacetoxy)cyclopentyl)amino-5,6-dimethyl-2-phenyl-7H-pyrrolo|(2,Za|pyrimidine , MS(EN): (MT) -380.1.

It should be noted that the following compounds can be obtained by the methods described above: 4-(cis-3-hydroxycyclopentyl)amino-5,6-dimethyl-2-phenyl-7H-pyrrolo (2,34) pyrimidine, MS(ЕН): (М -1)-323.1. salt of trifluoroacetic acid and 4-(cis-3-(2-aminoacetoxy)cyclopentyl)amino-5,6-dimethyl-2-phenyl-7H-pyrrolo|2,Za|pyrimidine, MS(EN): (MT) - 380.1 4-(3-acetamido)piperidinyl,-5,6-dimethyl-2-phenyl-7H-pyrrolo|2,Za|-pyrimidine, MS(EN): (M -1)-364.2. 4-(2-M'-methylcarbamidopropyl)iamino-5,6-dimethyl-2-phenyl-7H-pyrrolo|2,34| pyrimidine, MS(EN): (M --1)-353.4. 4p(2-acetamidobutyl)amino-5,6-dimethyl-2-phenyl-7H-pyrrolo|2,3a|-pyrimidine, MS(EN): (M -1)-352.4. then 4-(2-M'-methylcarbamidobutyl)amino-5,6-dimethyl-2-phenyl-7H-pyrrolo-(2,3a|pyrimidine, MS(EN): (M"-1)-367.5. 4-(2-aminocyclopropylacetamidoethyl)amino-2-phenyl-7H-pyrrolo-(2,34|pyrimidine, MS(EN): (M --1)-309.1. 4-(trans-4-hydroxycyclohexyl)amino -2-(3-chlorophenyl)-7H-pyrrolo-(2,3a|pyrimidine, MS(EN): (M --1)-342.8. 4-(trans-4-hydroxycyclohexyl)amino-2-( 3-fluorophenyl)-7H-pyrrolo-(2,3a|pyrimidine, MS(EN): (M-1)-327.2. 4-(trans-4-hydroxycyclohexyl)amino-2-(4-pyridyl)- 7H-pyrrolo-(2,3a)pyrimidine, MS(EN): (M7--1)-310.2.

Example 17

Scheme of IX with si a

So - go--ADO

K B mit ay » st s t" s I" soi a y v, bro "o sch

Mr. and Mrs

SA, gro o and So"

Nitrogen of pyrrole (7) (Scheme IX) was protected with di-tert-butyldicarbonate under basic conditions to obtain (Ce) the corresponding carbamate (22). Radical bromination of compound (22) proceeded regioselectively with the formation of co bromide (23). Basically, compound (23) is a key electrophilic intermediate for various nucleophilic coupling partners. Replacement of alkyl bromide with sodium phenolate trihydrate leads to the preparation of compound (24). Further replacement of the aryl chloride and removal of the tert-butylcarbamate protecting group in one step of the reaction leads to the desired compound (25).

A detailed description of the synthesis of compounds (22)-(25) in accordance with Scheme IX - p

Co-; gr Lo - s go :z» "7

Di-tert-butyl dicarbonate (5.37 g, 24.6 mmol) and dimethylaminopyridine (1.13 g, 9.2 mmol) were added to the solution,

THAT contains compound (7) (1.50g, 6.15mmol) and pyridine (30ml). After 20 hours, the reaction mixture was concentrated and the -I residue was partitioned between СНоСи» and water. The СНоSiO» layer was separated, dried over NaOH), filtered and concentrated to obtain a black solid. Flash chromatography (SO 5; 1/9 EtOAc/hexane, 0.40) afforded 1.7 g (80965) of a white solid (22). "H-NMR (200 MHz, SOSIv) 85: 8.50 (m, 2H, Ag-H), 7.45 (m, ЗН, Ag-H), -I 6.39 (s, 1H, pyrrole- H), 2.66 (s, ZN, pyrrole-CH"), s 20 s

FO mountain "M ko 23 - 60 1.76 (c, 9H, carbamate-CH"); MS, Man1- 344.1; T. pl. 175-1779С,

M-Bromosuccinimide (508mg, 2.8bmmol) and AIBM (112mg, 0.68mmol) were added to a solution containing compound (22) (935mg, 2.71mmol) and SCI) (50ml). The solution was heated under reflux. After 2 hours, the reaction mixture was cooled to room temperature and concentrated in vacuo to give a white solid.

After flash chromatography (50 5; 1/1 СНоСи/hexane, K-О0.30) 9bOmg (8495) of a white solid was obtained (23). "H-NMR (200 MHz, SOSIv) 85: 8.52 (m, 2H, Ag-H), 7.48 (m, ЗН, Ag-H), 6.76 (s, 1H, pyrrole-H) , 4.93 (s, 2H,

pyrrole-CH.VH), 1.79 (c, ZN, carbamate-CH3z); MS, M1-423.9; T. pl. 155-15726,

SI rum

MM s oto m 7

Sodium phenoxide trihydrate (173 mg, 1.02 mmol) was added in one portion to a solution of bromide (23) (41 mg, 0.97 mmol) in SiOCl (5 mL) and DMF (1 mL). After 2 hours, the reaction solution was partitioned between СНоСи» and water.

The water layer was extracted by SNSI. The combined СНоСИ»-layers were washed with water, dried with Mo5zO, filtered and concentrated to obtain a yellow solid. Flash chromatography (5% 25; 1/9 EtOAc/hexane, -0.30) afforded 210 mg (50965) of a white solid (24). "H-NMR (200 MHz, SOSIv) 85: 8.53 (m, 2H, Ag-H), 7.48 (m, 3H, AgH-H), 7.34 (m, 2H, Ag-H) . -SNuz); MS, M--436,2.

U-o n-M

In the school

C in (o) 25

A solution containing compound (24) (85 mg, 0.20 mmol), M-acetylethylenediamine (201 mg, 1.95 mmol) and DMSO (3 mL) was heated to 1002. After 1 hour, the temperature was raised to 1302. After 3 hours, the reaction the mixture "0 was cooled to room temperature and distributed between E(OAc) and water. The aqueous layer was extracted with E(OAc (2X).

The combined BEFOAs layers were washed with water, dried over NaCl, filtered and concentrated. After flash chromatography (5%, 110 EUN/SNCI3, VI-0.25), 7 mg (9396) of a white foamy solid (25) was obtained. "H-NMR (200 MHz, DMSO-ydv) 5: 11.81 (width s, 1H, M-H), 8.39 (m, 2H, Ag-H), 8.03 (width t, 1Н, М-Н), 7.57 s (width, 1Н, М-Н), 7.20-7.50 (m, 5Н, AG-Н), 6.89-7.09 (m , ZN, Ag-H), 6.59 (s, 1H, pyrrole-H), 5.12 (s, 2H, AgCH»O), 3.61 (m, 2H, MH»), 3.36 ( m, 2H, MON"), 1.79 (s, ZN, SOSN"); MS, M-1-402.6. -

The following compounds were obtained by the method described in Example 17: 4-(2-acetylaminoethyl)amino-6-phenoxymethyl-2-phenyl-7H-pyrrolo(2,3a|pyrimidine, m.p. 196-197; MS(EN): 401.6 (M'--1). "20 4-(2-acetylaminoethyl)amino-6-(4-torphenoxy)methyl-2-phenyl-7H-pyrrolo|2,34), pyrimidine. MO(EN)Y 4201 Z s (M). and 4-(2-acetylaminoethyl)amino-6-"4-chlorophenoxy)methyl-2-phenyl-7H-pyrrolo|2,34| pyrimidine. MOSC(EN) 4361 "" (M"). 4-(2-acetylaminoethyl)amino-6-(4-methoxyphenoxy)methyl-2-phenyl-7H-pyrrolo |2.34| pyrimidine MS(EN): 432.1 45. (M), - 4-(2-acetylaminoethyl) amino-6-(1,1-pyridin-2-one) methyl-2-phenyl-7H-pyrrolo (|2, 34 | pyrimidine. MOS(EN): 403.1 7 (M). 4-(2-acetylaminoethyl)amino-6-(M-phenylamino)methyl-2-phenyl-7H-pyrrolo|2, ZaIpyrimidine. MS(EN ): 400.9 (M-1). - 4-(2-acetylaminoethyl)amino-6-«"M-methyl-M-phenylamino)methyl-2-phenyl-7/H-pyrrolo |2,34| pyrimidine Me(EN): p» 070 d1AB(M).

Ф 4-(2-M'-methylcarbamidoethyl)amino-b-phenoxymethyl-2-phenyl-7H-pyrrolo|2.34| pyrimidine MS(EN): 416.9 (M"--1).

Analysis of human adenosine receptors A and Aba using the yeast p-galactosidase reporter gene

Yeast strains (5. segemisiae) were transformed with human adenosine AT (AIK; SABO strain SU12660) or human Aga (Aga; SABO strain SMU12660) and the Ias7 p-galactosidase reporter gene was added for (F. use as a functional index. A full description of the transformations is given below (see the section "Yeast strains. For all analyses, MESA (5'-M-ethylcarboxamidoadenasine), a strong adenosine receptor agonist with similar affinity for A1 and Aga receptors, was used as a ligand. The tested compounds were evaluated at 8 concentrations (0 ,1-10000 nM) on their ability to inhibit MESA-induced b-galactosidase activity under the influence of SU12660 or SU8362.

Preparation of initial yeast cultures. Each of the corresponding yeast strains, SU12660 and SU8362, was streaked onto an I T-agar plate and incubated at 302C until colonies appeared. Yeast from these colonies was added to I T-liquid (pH 6.8) and cultivated overnight at 302С. Each yeast 65 strain was then diluted to OPeoo-1.0-2.0 (approximately 1-2x107 cells/ml) as determined by spectrophotometry (Moiiesciag Oemisez MMAH). 4 ml of 4095 glycerol (1:1.5 vol.:vol.) was added to each bml of yeast liquid culture.

("yeast/glycerin mother liquor"). Ten 1ml aliquots were taken from this yeast/glycerol mother liquor and stored at -802C until required for analysis. Analysis of yeast receptors A1 (A.K) and Aga (Ag2ak). One vessel containing the royal yeast/glycerin solution for each of SU12660 and

Su8362, melted and used for inoculation of supplemented liquid I T medium, pH 6.8 (92 ml) of liquid

And T, to which was added: 5 ml of 4095 glucose, 0.45 ml of 1 M KOH and 2.5 ml of Rirevz, pH 6.8). Liquid cultures were grown at 302C for 16-18 hours (overnight). Then, aliquots taken from overnight cultures were diluted in the medium

IT containing 4 units/ml of adenosine deaminase (type MI or MII, obtained from the intestinal mucosa of a calf, zZidta) with the production of OP v0o0-0.15 (1.5x106 cells/ml) for SU18362 (Agak) and OP 600 -0.50 (5x106 cells/ml) for 70. СХ12660 (А1К).

Analyzes were performed with a final volume of 100 μl in 9b-well microtiter plates, so that a final concentration of 295 DMSO was reached in all wells. For primary screening, 1-2 concentrations of the tested compounds (10 μM, tμM) were used. 8 concentrations (10000, 1000, 500, 100, 50, 10, 1 and 01 NM) were tested to obtain the profile of the compounds. For each microtiter plate, 1 µM of 2095 DMSO was added to the "control" and "full" wells 75, and 1 µM of the test compound (in 2096 DMSO) was added to the "unknown" well. Then 1 µM of MESA (5 mM for ATK, 1 µM for Agac) was added to the "full" and "unknown" wells, and 1 µM of PB5 was added to the "control" well. At the final addition, 80 μl of yeast strain SU8362 or

SU12660. All tablets were then quickly mixed (on a laboratory orbital shaker I ab ipe for 2-3 minutes) and left to incubate for 4 hours at 302С in a dried oven. 20 p-Galactosidase activity can be estimated using colorimetric (for example, OMRO,

SPO), luminescent (for example, Cyasiop-Ziag or fluorimetric substrates (for example, RBOO,

Kezogype). Currently, fluorescent detection is preferred, which is based on a higher signal-to-noise ratio, which is basically free from interference and is inexpensive. Fluorescein degalactopyranoside was added to all wells at a concentration of 2 µM/well (final concentration - 8 µM). The plates were shaken for 5-6 seconds (on a Gary Ipe laboratory orbital shaker) and then incubated for 9 hours. at 3720 (9595 05 590 CO. incubator). After the end of the 90-minute incubation period, the development of β-galactosidase activity was stopped using 20 μl/well of 1 M Ma 2 CO»z, and all tablets were shaken for 5 seconds. Then the tablets were shaken for bsec. and determined the relative intensity of y-oi 30 fluorescence using a fluorimeter (Tesap Zresigaysnog; excitation - 485 nm, emission - 535 so nm).

Calculation. The relative fluorescence value for the "control" well was interpreted as background and subtracted from the values for the "full" and "unknown" wells. Profiles of compounds were analyzed using a logarithmic He transformation (x-axis: compound concentration), and then to calculate values of ISvo (SsgarpRaai Rgizt).

From yeast strains. The strains Zasspagotusev segemiziae SU12660 Gagi"1442 160Й-1 Ty8I-NIBZ sapi t vie14: ris! 52 vie3"1156 drai (41) -SsiZ Iuv2 igaz Iets2 (gri:piz3; GEO2 ROCr Mio eadeg-PAIK-RNOBegt 2ti-ogid) were obtained

KERZ Atrig| and SU8362 (|draIr--b55BIOK tTagi"1442 1B60И-1 7to5I-NIZZ sapi vie14:grI! 52 v8ie3"1156 Iuv2 gas

Iets2 Igri piz3; I 62 ROCr-pAgak 2ty-ogi KERZ Atrg|). « 20 Medium I T. Medium I T (I eny-Ttr-supplemented) consists of 100 g of the yeast nitrogenous base of OIESO and the following components added to it: 1.0 g of valine, 1.0 g of aspartic acid, 0.75 g of phenylalanine, 0 ,9g of lysine, 0,45g of tyrosine, 0,45g of isoleucine, 0,3g of methionine, O,bg adenine, O0,4g of uracil, O0,Zg serine, 0,3g:z» proline, 0,Zg of cysteine, 0, 3g of arginine, 0.9g of histidine and 1.0g of threonine.

Construction of yeast strains expressing human AT adenosine receptor

This example describes the construction of yeast strains expressing human adenosine receptor A1 -1, functionally integrated into a cascade of reactions involving pheromone in the yeast system.

I. Construction of the expression vector ko To construct a yeast vector expressing the human adenosine receptor AI, cDNA of the AT adenosine receptor was obtained by PCR using the reverse transcriptase me NC of the human hippocampus using primers designed on the basis of the published sequence (95) of the AT adenosine receptor human and with the help of standard equipment. The PCR product was subcloned into Meso!-i

F Khwai!-sites expressing yeast plasmin pMP 15.

Plasmid PMP15 was obtained from ri RHI as follows: HraY-site MERBI1 (Vgoasi, U.K. ei ai. (1983) "Mesioge Tog podp-Ieme!i, ipadisirbie oehrzhezviop oi osiopedy depez ip ueazi r.83-117 ip M. Ipotsue (Eid.), Ehregitepia! 5 Mapirshiayop oi Sepe Ehrgezviop. Asadetis Rgezv, Mem/ Mogk) were eliminated by hydrolysis, completed at the ends and ligated again to give Herb51MsoYurHra. Another HraI site was constructed in the VatNi site by (F) Watni restriction , completed at the ends, ligated with a linker (Mem Epodiapa Vioiarv, 21081), cleaved by Hbai

Ge and ligated again to give MERBIMsoHi. This plasmid was cleaved by Ezvr3 and Msoi and ligated with the fragments

Yets2 and ROKr generated by PCR. 2 t.p.o.-en2-PCR-product was generated by amplification with ВоЖЕРЗИМсСОо using primers containing Ezr31- and Во. The PCR product of rOKr, consisting of 600 base pairs, was generated by amplification from rOKase (Kapo, M.-5. ei aiYi. (1990) Moi.SeiI.VioYi. 10:2582-2590) using PCR primers that contain VaiSh and MsoI sites. The resulting plasmid was labeled with ri RHI. ri RHI was modified by inserting the coding region of the α-factor leader pre-pro sequence into the MeOi site.

The prepro leader sequence was inserted so that the McoI cloning site was retained at the C"-end of this leader sequence, but not restored at the 5-end. Thus, the receptors can be cloned by cleavage of the plasmid with the enzymes McoI! and Chrai. The resulting plasmid was designated pMP15.

Plasmid PMP15, in which cDNA of human AT adenosine receptor was integrated, was designated rbooO95. In this vector, the cDNA of the receptor was ligated to the C3-end of the prepro-leader sequence of the yeast α-factor. In the process of protein maturation, peptide prepro-sequences were cleaved with the formation of a mature full-length receptor. This occurs during the processing of the receptor by means of a cascade of yeast secretory reactions.

This plasmid was maintained by I ei-selection (ie, by cultivation on a medium containing no leucine). The sequence of the cloned coding region was determined and found to be equivalent to the sequence published in the literature (SepVapk accession numbers 545235 and 556143).

II. Construction of yeast strain 70 To create a yeast strain expressing human AT adenosine receptor, yeast strain SU7967 was used as a parent strain. SU7967 has the following genotype: MATa drab1163 dra1(41)saiZ

Tag101442 1-1 RHO51-NIZZ sapi viei4: gri: 52 eiezZriI 156 Iuz2 igaz Iets2 gri piz3.

Genetic markers are presented below:

MATA............... Crossbreeding type a.

Sra101163.........Endogenous SORA1 Yeast O-protein deleted.

Сра1(41)Саи3......dra1(41)-СаиЗ is integrated into the yeast genome. This chimeric Са protein consisted of the first 41 amino acids of the endogenous СА subunit; yeast Ca protein attached to the mammalian C-protein Cy3, in which the cognate M-terminal amino acids were deleted.

ЕБатОо1442........the BAKI gene (responsible for the termination of the cell cycle) was delegated (as a result of which the cell cycle is terminated after the activation of the cascade of reactions of the pheromone response).

Tri-1............ strain with high transformation efficiency by electroporation.

EOB1-NIZZ........hybrid of the GROBI promoter with the coding region (resulting in the creation of the pheromone-inducible NIZZ3 gene).

Sap1.............. a rginine/canavinine permease. high school

Zie14:gr1:1u52....disruption of the 5TE14, C-farnesylmethyltransferase gene (due to which the transmission of the basal signal via the pheromone pathway decreases). i) vie3O1156........-endogenous yeast 5TK, factor a pheromone receptor (ZTEZ) was destroyed.

Iuz2............. defective in 2-aminoapidate reductase, yeast, for their growth, lysine is necessary.

MhaZz.............defective for orotidine-5'-phosphate decarboxylase, yeast, for their growth, HezoUuracil is necessary.

Ie2.............defective in p-isopropylmalate dehydrogenase, yeast, for their growth, leucine is necessary. and

Tgr11...........defective for phosphoribosyl-lanthranilate, yeast, for their growth, tryptophan is necessary. M half 3............defective for imidazole glycerol phosphate dehydrogenase, yeast, for their growth, histidine is necessary. with

In strain SU7967, by electroporation, two plasmids were transformed: plasmid p095 (encoding the human α-adenosine receptor At, described above) and plasmid p1584, which carries the GOBI-P-galactosidase reporter gene.

Plasmid p1584 was obtained from plasmid pKk5426 (SpNgiziapsop, T.MU. ei a), (1992) Sepe 110:119-1122). Plasmid pK5426 contains a polylinker site in nucleotides 2004-2016. To obtain plasmid ri 584, promoter hybrid

ROBI1 genes of the b-galactosidase gene were inserted into the Eadi and Khpoi restriction sites. Plasmid rib584 was maintained by Tgr selection (ie, by culturing on leucine-free medium). with c The obtained strain, carrying rbBO95 and ri584 and labeled SU12660, expressed human adenosine

And receptor At. For the cultivation of this strain in a liquid medium or on plates with agar, it was used minimal media that do not contain leucine and tryptophan. To conduct an analysis of the growth of this strain on plates (analysis on PO51-NIZZ), plates were used with a medium at pH 6.8 containing 0.5-2.5 mM of 3-amino-1,2,4-triazole and that do not contain leucine, tryptophan and histidine. As a control for specificity, in all -I experiments, a screening was carried out with a comparison of seven transmembrane receptors originating from one or more other yeast species. o Construction of yeast strains expressing the human adenosine receptor Aga -I This example describes the construction of yeast strains expressing the human adenosine receptor 5p Aga, which was functionally integrated into a cascade of reactions involving the pheromone in the yeast system. o I. Construction of an expressing vector

Ф For the construction of a yeast vector expressing the human Aga adenosine receptor was obtained

cDNA of adenosine receptor Aga from Og. RpiYi Mygrpu (MIN). After obtaining this clone, its sequence was sequenced for the insertion of the Aa receptor and it was found to be identical to the published sequence of dv (ZepVapk accession number y 546950). The cDNA of the receptor was excised from this plasmid by PCR using the MEMT polymerase and cloned into the plasmid ri РВХ, in which the expression of the receptor was regulated

F) constitutive promoter of yeast phosphoglycerate kinase (POK). The sequence of the entire insert was then re-sequenced and found to be identical to the published sequence. However, with the strategy used here, three amino acids (SiuzZegua) were attached to the carboxy terminus of the receptor. in II. Construction of a yeast strain

To create a yeast strain expressing human Aga adenosine receptor, yeast strain SU8342 was used as a parental strain. SU8342 has the following genotype: МАТа яг101442 1-1 Иуз2-игаз

Ie2 igri piIv3 708I-NIZZ sap!i zie3z01156 drab1163 vieid:zhgr1:1ЮU52 aratr-gza5ETOK (or draitr-g4502295 or dratr--b85ET1OKO2295). 65 Genetic markers are described in Example 1 with the exception of C-protein variants. For expression of the human receptor

Also, yeast strains were used in which the endogenous CPA1 of the yeast protein C was deleted and replaced by Co. a mammal Three rat S o. mutants were used. These variants contain one or two point mutations that convert them into proteins that efficiently bind to yeast Du. They were identified as boeETOK (in which the glutamic acid at position 10 is replaced by lysine), b 20.02295 (in which the aspartic acid at position is replaced by sermine) and 50E1OKO2295 (which contains both point mutations).

Strain SU8342 (carrying one of the indicated three mutant rat So.-proteins) was transformed either with the parental vector ri РВХ (receptor") or ri РВХА?2a (receptor "1"). To assess the degree of activation of the pheromone response pathways, a plasmid with promoter RG9O51 attached to the sequences encoding p-galactosidase (described above).

Functional analysis using yeast strains expressing the human A1 adenosine receptor

In this example, the development of functional analysis by the method of screening in yeast for modulators of the human adenosine receptor A1 is described.

I. Ligands used in the analysis

Adenosine, a natural agonist for this receptor, and two other synthetic agonists were used to develop this assay. In a subseries of experiments, adenosine, which is known to have an ES of approximately 75 nM, and (-)-M6-(2-phenylisopropyl)adenosine (RIA), which is known to have an affinity of approximately 50 nM, were used. 5'-M-ethylcarboxamidoadenosine (MESA) was used in all growth assays. To prevent signal transmission due to the presence of adenosine in the culture medium, adenosine deaminase (40 d./ml) was added to all analyses.

I. Biological response in yeast

The ability of the AT adenosine receptor to functionally bind in a heterologous yeast system was assessed by introducing a vector expressing the A1 receptor (p5095, described above) into a series of yeast strains expressing different subunits of protein b. Most of these transformants expressed subunits of c o. with subtypes So or Bo. Additional Co proteins were also tested for the possible identification of non-specific (3) binding of the b o protein to the receptor. ZTE18 or the chimeric construct ZTE18-C 2 were integrated into the yeast genome of various strains. The yeast strains contained a defective gene of НИЗЗ and an integrated copy of РО5З1-НИЗЗ , which allows for selection in a selective medium, which contains 3-amino-1,2,4-triazole tested at 0.2, 0.5 and 1.0 mm) and which does not contain histidine. Transformants were isolated and monolayers were obtained on a medium containing 3-amino-1,2,4-triazole, 4 units/ml of adenosine deaminase and containing no histidine. Then five microliters of different ligand concentrations were applied (for example, MES A at 0, 0.1, 1.0 and 10 mM). Growth was monitored for 2 days. Thus, responses in the form of ligand-dependent growth were tested in different yeast strains. -

The results are systematized below in Table 1. The symbol (-) means that no ligand-dependent activation of receptor C was detected, and (i) means a ligand-dependent response. The term "ICMA" indicates ligand-independent receptor-mediated activation. hey

Also 017 vvm0001yazhv 10000000 - nnya 0011 - PI ns ss: NN po ps ;" ; ная 0001 кв 00000100 ямк чо rom p he I MY PO POI z St ovmtbyantyuyuny UE6O1000000 - o) 5 roms 11 ni t LEARN THE FIELD PO ON yy ni t SUMY NOYA PO ON

PO Ps NNYA PON VA zno U MY PO POI o p U AND MY PO POI ps: WE p Ps NNYA NANNYA and Roma 11 ni t TEACHING FIELD PO ON 60 ni t SUMY NOYA PO ON

PO Ps NNYA PON VA no U MY PO MY p he AND MY FIELDS POI

As shown in Table 3, stable transmission of most signals was found to occur in the yeast strain expressing the SORAT1 (41)-Co;3 chimera.

I. Analysis on Tyv1-I as/

For a more complete characterization of the activation of the pheromone response cascade, the synthesis of p-galactosidase was evaluated using Giz1-I as in response to agonist stimulation. To perform an analysis of p-galactosidase, increasing concentrations of human adenosine receptor AT expressed in a yeast strain co-expressing the chimera 5ie18-5u2 and 70. ЗРА,4-Со;з were added to the one in the middle phase of logarithmic growth of the culture the ligand. Transformants were isolated and cultivated overnight in the presence of histidine and 40 units/ml of adenosine deaminase. After b-hour incubation with 40 units/ml of adenosine deaminase and ligand, induction of b-galactosidase was measured using SRKO as a substrate for b-galactoside. 5x105 cells were used in each analysis.

The results obtained during the stimulation of MESA indicated that at a concentration of MESA of 108 M, an approximately 2-fold increase in the level of stimulation of p-galactosidase activity is achieved.

Moreover, at a concentration of MESA of 105 M, an approximately 10-fold increase in the level of stimulation was observed.

The effectiveness of this assay was increased by confirming the activity of antagonists against this strain. Two known adenosine antagonists, KAS and ORSRH, were tested for their ability to compete with MESA (at 5mM) for activity in the β-gadactosidase assay. In these assays, 7 induction of D-galactosidase was measured using ROS as a substrate and 1.6 x 105 cells per assay.

The results showed that both KAS and UORSRKH are strong antagonists of adenosine receptor A1 expressed in yeast with IC values of 44 nm and 49 nm, respectively.

In order to determine whether this inhibitory action is specific to the At subtype, a series of additional experiments was conducted by analyzing the Aga receptor in yeast (described in Example 4).

The results obtained in an assay using Aga yeast showed that KAS was a relatively effective Aga receptor antagonist, consistent with published data. In contrast, ORSRH, as predicted from published data, was relatively inert to this receptor.

IM. Radioactive ligand labeling with 30 Adenosine receptor analysis, in addition, consists in measuring the binding parameters of radioligands for the receptor. Substitutional binding | ZNISRX with several known compounds that are ligands for the o adenosine receptor, HAC, ORSRX and COo5 were analyzed using membranes obtained from yeast M expressing the human adenosine receptor A1. To assess the specificity of binding, the results obtained using yeast expressing the human adenosine receptor At were compared with the results obtained for yeast expressing the human adenosine receptor Ada or the human adenosine receptor AZ. To perform this assay, 50 milligrams of membranes were incubated with 0.4 mM I ZNISRH and increasing concentrations of adenosine receptor ligand. Incubation was carried out in 50 mM Tris-HCl, pH 7.4, 1 mM EOTA, 10 mM Masi, 0.2596 VZA and 2 units/ml of adenosine deaminase in the presence of protease inhibitors for 60 minutes at room temperature. Binding was assessed by adding ice-cooled 5HOMM Tris-NSII, pH 40.74 and 10MM Mosi", followed by rapid filtration on SOR/B filters pre-impregnated with 0.590 - s polyethyleneimine, using a collection of Raskag 96-well tablets. The data were analyzed by constructing a nonlinear curve using the least squares method using Rgizt 2.01 software. "» The IS values obtained in this experiment are systematized below in Table 4:

F

These data indicate that the known compounds have an affinity similar to that reported in the literature. (iF) These data further demonstrate that the yeast assay has sufficient sensitivity to distinguish receptor subtype specificity.

Functional analysis using yeast strains expressing the human adenosine receptor bo Ag

In this example, the development of functional analysis by the method of screening in yeast for modulators of the human adenosine receptor A1 is described.

I. Ligands used in the analysis

The natural ligand adenosine, as well as other well-characterized and commercially available ligands, were used to study the human Aga adenosine receptor functionally expressed in yeast 65 . Three ligands were used to perform this analysis. These ligands are:

Ligand Known Key, Function

Adenosine BOoOnM agonist 5-M-ethylcarboxamidesadenosine (MESA) 10-15NM agonist (-)-m6-(2-phenylisopropyl)adenosine 100-125nM agonist (RIA)

In order to prevent transmission of the signal due to the presence of adenosine in the culture medium, adenosine deaminase (4 units/ml) was added in 70 all analyses.

II. Biological response in yeast

Agonists of the Aga adenosine receptor were evaluated for their ability to stimulate the pheromone response cascade in yeast transformed with a plasmid expressing the Aga receptor and expressing or

So5E1OK, or So.,02295, or bo5E1OK02295. The ability of the ligand to stimulate the pheromone 7/5 response cascade in a receptor-dependent manner is indicated by an alteration in the yeast phenotype. Activation of the receptor led to the modification of the histidine auxotroph phenotype into a histidine phototroph (Ty8I-NIZZ activation). Three independent transformants were isolated and cultured overnight in the presence of histidine. Cells were washed to remove histidine and diluted to a concentration of 2 x 109 cells/ml. μl of each transformant was spotted onto non-selective media (including histidine) or selective media (1 mM AT) in the absence of 2o or in the presence of 4 units/ml of adenosine deaminase. Tablets were cultured at 302C for 24 hours. In the presence of histidine, both the receptor "strain (BK) and the receptor strain (B) were able to grow. However, in the absence of histidine, only K "7 was able to grow. Since no ligand was added to these tablets, this fact can have two explanations. One possible interpretation of this fact is that yeast carrying the receptor has preferential growth due to ligand-independent activation mediated by the SM receptor ("/IVEMA"). Alternatively, yeast itself can synthesize the adenosine ligand. To detect these two possibilities, an enzyme that decomposes the ligand adenosine deaminase (ADA) was added to the growing yeast and to the tablets. In the presence of adenosine deaminase, K"-cells stopped their growth in the presence of histidine, which indicated that the yeast really synthesizes the ligand.

This interpretation was confirmed by analysis of A2a growth in liquid medium. In this experiment, uh-oh

KE yeast (strain Co8ETOK, expressing the Aga receptor) was inoculated at three densities (1x10 cells/ml; 3x109 cells/ml; or 1x105 cells/ml) in the presence or absence of adenosine deaminase (4 units/ml) . The rigidity of the analysis increased with increasing concentration (0.01, 0.2 or 0.04 mM)

Z-amino-1,2,4-triazole (AT), a competitive antagonist of imidazoleglycerol-P-dehydratase, a protein product of the NIBZZ gene. In the presence of adenosine deaminase and 3-amino 1,2,4-triazole, the yeast grew more slowly. However, in the absence of 3-amino-1,2,4-triazole, adenosine deaminase had little effect. Thus, adenosine deaminase itself does not have a direct effect on the cascade of reactions of the pheromone response.

An alternative way to measure growth, and one that can be scaled down for more thorough screening, is an Aga receptor ligand spot assay. The bo5ETOK strain, expressing the Aga receptor (Agayutk"), or the strain that does not contain the receptor (KU), was cultured overnight in the presence of histidine and - c d4od./ml adenosine deaminase. Then the cells were washed to remove histidine and diluted to concentration "» 5Bx1059 cells/ml. 1x109 cells were plated by spreading on plates with a selective medium containing 4 units/ml adenosine deaminase and 0.5 or 1.0 mM 3-amino-1,2,4-triazole (AT) and left for 1 hour for drying

Then, the following reagents were applied to the monolayers in 1 ml; 1 mM oadenosine, 38.7 mM histidine, dimethylsulfoxide (DMSO), 10 mM RIA or 10 mM MESA. Cells were cultured at 30°C for 24 hours.

The results showed that without the receptor, these cells could grow only in the presence of histidine added to the medium. In contrast to this, K' cells grew only in those areas where RIA and MESA ligands were stained. Since these plates contained adenosine deaminase, the absence of growth where adenosine was spotted confirmed adenosine deaminase activity. at 250 IP. Analysis on -Ti51-I as/

For the quantitative characterization of the activation of yeast crossing pathways, the synthesis of γ-galactosidase was evaluated using the Ty851-(assay. Yeast strains expressing 0-E1OK, bo.02295 or bo8E1OKnO2295 were transformed with a plasmid encoding the human receptor Aga (K"), or a plasmid, which lacked this OV receptor (K). Transformants were isolated and cultured overnight in the presence of histidine and 4 units/ml of adenosine deaminase. 1x10' cells were diluted to a concentration of 1x109 cells/ml and exposed to growing

IF) concentrations of MESA for 4 hours, and then D-galactosidase activity in these cells was determined. where The results showed that K/ - strains mainly did not show p-galactosidase activity, while B' - strains expressing b20-ETOK, bo.02295 or bo5E1OKnO2295, showed increasing amounts of p-galactosidase 60 activity as the concentration increased MESA, which indicated that in response to exposure to increasing concentrations of the ligand, a dose-dependent increase in β-galactosidase units is observed. This dose dependence was observed only in cells expressing the Aga receptor. In addition, the most active О о -construction in relation to the Aga receptor was the construction З ОЕТЛОК. The construction bo.02295 was the second most active in relation to the Aga receptor, and of these three tested constructions, the construction o5ETOKO2295 was the least active in relation to the Aa receptor, however, even the construction boE10K02295 stimulated easily detectable amounts of β-galactosidase activity .

For a more complete description of the identified analyses, see US Patent Application Reg.Mo09/088985, entitled "Ropsiiopa! ehrgeziop oi adepozipe Keseriogz ip Ueazi", and filed on June 2, 1998 (Patent Register MoSRI-093), the full content of which is incorporated into this description by reference.

Pharmacological characterization of human adenosine receptor subtypes

Materials and methods

Materials. ("NI- ORSRKH|Cyclopentyl-1,3-dipropylxanthine, 8-|Idipropyl-2,3-3H (M) | (120.0Ky/mmol); IZNI - so5 21680, Icarboxyethyl-(M) / ZOKy/mmol ) and HOSPYADAV-MESA (00 -4-aminobenzyl-b' 70 -M-methylcarboxamidoadenosine) (2200Ki/mmol) were supplied by Mem Epdiapa Mysiveag (Vosiop, MA).

HAC (an analogue of xanthineamine); MESA (5'-M-ethylcarboxamidoadenosine); and IVA-MESA were supplied by Kezeagsp

Viospetisa!in Ipiegpaiopa! (KVI, Maiysk, MA). Adenosine deaminase and tablets of a complete mixture of protease inhibitors were supplied by Voeyipdeg Mapppeit Sogr. (Ipaiaparoiiv, IM). Membranes from NEK-293 cells stably expressing subtypes of human adenosine receptor 2a (KV-NAZha); adenosine receptor 75.265 (KV-NA2BI|) or adenosine receptor C (KV-NAZI), respectively, were supplied by Keseriog Viiodu (Veyivmshe, Moscow State). Cell culture reagents were supplied by I. Ye Tesppoiodiev (Sgapa Iviapa, Moscow State), except serum, which was supplied by Nusiope (I odap, OT).

Yeast strains. Strains bZasspagotusevs segemivide SU12660 Mag1"71442 16C-1 Ty851-NIBZ sap!i vie14siri:1іM52 vie3"1156 dra1(41)-boi3Z Iuz2 tsgaZ Iei2 igri/yig3; GEO2 ROKr-Me/1I eadeg-pATK-RNOBiegt 2ti-ogid KERZ Atrg)| and Sm8362 Idrair-gazazEi1OK tTag-"1442 6-1 Ty8I-NIZZ sapi vie14:zhgr1i1MU52 vie3"1156

Iuz2 igaz Iets2 igri piz3; GEO2 ROKr-pAgak 2ty-ogi KERZ Atrg| were obtained as described above.

Yeast culture. The transformed yeast was cultivated in a medium with I ey-Tgr | T) (pH 5.4), to which 295 g of glucose was added. To obtain membranes, 250 ml of medium I T was inoculated with an initial titer of 1-2 x109 cells/ml from ZOml of overnight culture and incubated at 302 with continuous oxygen supply and vortex CM 29 mixing. After 16-hour cultivation, the cells were collected by centrifugation and obtained he) membrane as described below.

Mammalian tissue culture. NEK-293 cells stably expressing human adenosine receptor subtype 2a (clone Sadiz 55) were cultured in Dulbecco's minimal maintenance medium (OMEM) supplemented with 1095 fetal bovine serum and their penicillin/streptomycin under selection pressure using ke,

BoOmg/ml antibiotic 5418 at 372C in an atmosphere of 596 CO" with increased humidity. co

Preparations of yeast cell membranes. After overnight incubation, 250 ml of culture were collected by centrifugation at 2000 rpm in a Bogmay centrifuge! KT6000. Cells were washed in ice-cooled water, - centrifuged at 492C, and the sediment was resuspended in 1 Oml of ice-cooled lysis buffer |bmMM s

Tris-HCI, pH 7.55 5MM EOTA; and 5 mM ESTAY, to which tablets with a mixture of protease inhibitors were added (1 tablet per 25 ml of buffer). Glass spheres (17g; 400-600 mesh; BZidta) were added to the suspension, and these cells were disrupted by vigorous vortexing at 42C for 5 minutes. The homogenate was further diluted

3 ml of lysis buffer containing protease inhibitors and centrifuged at 3000 x for 5 min. Then the membranes were deposited at 36,000 ha (Zogmai! KO5B, rotor type 5534) for 45 minutes. The resulting membrane precipitate was resuspended in 5 ml of membrane buffer (50 mM Tris-HCII, pH 7.5; 0.6 mM EOTA; and 5 mM Mosig2), to which tablets from a mixture of protease inhibitors were added (1 tablet per 50 ml of buffer) and stored at -809C for further experiments. "" Preparation of membrane preparations of mammalian cells. Membranes of NEK-293 cells were obtained as previously described (Oilis E. ei ai., 9U.Vioi.Spet., 267, 9844-9851, 1992). For this, the cells were washed with PB5 and collected with a rubber stick. Cells were deposited by centrifugation at 200 rpm at 42C on a Zogmai centrifuge! KT6000. Pellet -I was resuspended in 5 ml/well in lysis buffer at 42 (5 mM Tris-HCl, pH 7.5; 5 mM EOTA; 5 mM ESTA; 0.1 mM phenylmethylsulfonyl fluoride, 1 ug/ml pepstatin A; and 10 ug/ml aprotinin) and homogenized in a homogenizer

Downs. Then the cell lysate was centrifuged at 36,000 x 49 (Zogmai! KO5V, rotor type 5534) for 45 minutes - and the sediment was resuspended in 5 ml of membrane buffer | 50 phenylmethylsulfonyl fluoride, 1 ug/ml pepstatin A; and 1 ug/ml alrotinin) and stored at -809С for further experiments.

I) To determine the total protein concentration in yeast and mammalian membranes, Vio-Kai protein analysis kits were used based on the Bradford dye binding procedure (Vgadioga, M, Apai!. Viospet. 72:248 (1976)). 22 Adenosine receptor subtype 1 saturation and competition for binding to a radioactive ligand

Gee! Binding with saturation and competitive binding on membranes obtained from yeast cells transformed with the human receptor subtype A1 was carried out using the antagonist I-NIORSCRH as a radioactive ligand. Membranes were diluted in binding buffer (HOMM Tris-HCI, pH 7.4; 10 mM Masig, 10 tt EOTA; 0.2595 B5A; 2 units/ml of adenosine deaminase and tablets of a mixture of protease inhibitors 1/50 ml | at 60 concentrations 1 ,0mg/ml When binding to saturation, the membranes (5µg/well) were incubated with increasing concentrations of |ZNIORSRH (0.05-25nM) in a final volume of 100µl of binding buffer at 252 for 1 hour in a 96-well microtiter plate in the absence and presence of 10 μM unlabeled HAC.

In competitive binding, membranes (5 µg/well) were incubated with |(-NIORSRH (1.0 nM) in a final volume of 5 10 µl of binding buffer at 252C for 1 hour in a 9b-well microtiter plate in the absence and presence of 100 µm of unlabeled KAS or with increasing concentrations of competing compounds.

Competitive binding of adenosine receptor subtype 2a with a radioactive ligand

Competitive binding on membranes obtained from HEK-293 cells stably expressing the human receptor of the Aga subtype was carried out using the agonist | ZNISO8-21680 as a radioactive ligand.

Membranes were diluted in a binding buffer (500 mM Tris-HCl, pH 7.4; 10 mM Mosi2, 1.0 mM EOTA; 0.2595 VA; 2 units/ml of adenosine deaminase and tablets of a mixture of protease inhibitors 1/500 ml| at concentrations 0.2 mg/ml.

Membranes (1 µg/well) were incubated with (H|ISO5-21680 (100 nM) in a final volume of 100 µl binding buffer at 259 for 71 h in a 9b-well microtiter plate in the absence and presence of 500 µM unlabeled MESA or with growing concentrations of competing compounds.

Competitive binding of adenosine receptor subtype C with a radioactive ligand

Competitive binding on membranes obtained from NEK-293 cells stably expressing the human receptor of the AZ subtype was carried out using the agonist 221) AB-MESA as a radioactive ligand. Membranes were diluted in binding buffer (bOMM Tris-HCl, pH 7.4; 10 mM Macia, 1.0 mM EOTA; 0.2595 VBA; 2 units/ml of adenosine deaminase and tablets of a mixture of protease inhibitors 1/50 ml | at concentrations 0.2mg/ml. Membranes 12 (10omkg/well) were incubated with I"22I) AB-MESA (0.75NM) in a final volume of 100μl binding buffer at 252 for 1 hour in a 9b-well microtiter plate in the absence and in the presence of 10 μM unlabeled

IV-MESA or with increasing concentrations of competing compounds.

At the end of the incubation, assays for the binding of receptors of subtypes AT, Aga and AZ with the radioactive ligand were completed by adding an ice-cooled buffer of 50 mM Tris-HCl (pH 7.4), to which 10 mM MacI2 was added, followed by rapid filtration through glass fiber filters ( 96-well filters SR/B

Opiginege, Raskaga) that were previously soaked in 0.595 polyethyleneimine in a cell collector Rikegtäie 196 (Raskaga). The filter tablets were dried, sensitized with 1 ml/well of scintillation liquid (Misgossipi--0, Raskaga) and counted in a TorSocpi counter (RasKaga). Analyzes were performed in three duplicates. In the AK, Agac, and AZK binding assays, nonspecific binding was 5.6--0.595, 10.8-4-1.490 s, and 15.1--2.695 of total binding, respectively. Gee)

Competitive binding of adenosine receptor subtype 26 with a radioactive ligand

Competitive binding on membranes obtained from NEK-293 cells, stably expressing the human receptor of the A2Bb subtype, was carried out using the antagonist of the AT receptor |ZNIORSRKH as a radioactive "ozo ligand." Membranes were diluted in binding buffer (1 mM Nerez-KOH, pH 7.4, containing 1.0 mM EOTA; 00.1 mM benzamidine and 2 units/ml of adenosine deaminase | at concentrations of 0.1 mg/ml. Membranes (15 μg /well) were incubated with I-NI ORSRH (15NM) in a final volume of 100μl of binding buffer at 252С for 1 hour in a 96-well microtiter plate in the absence and presence of 10μM of unlabeled HAC or with increasing concentrations of competing compounds At the end of the incubation, the analysis was completed by adding 3.5 ml of ice-cooled 10 mM Nerez-KOH buffer (pH 7.4) followed by rapid filtration through glass fiber filters (96-well filters SOB/SOpiginegv5, RasKaga) previously soaked in 0.590 polyethyleneimine in a Rikeghtaie 196 cell harvester (RasKaga). The filter tablets were dried, sensitized with 5µl/well scintillation fluid (Misgobsipi-20, Raskaga) and counted in a counter "

TorSotschpi (Raskaga). Analyzes were performed in three duplicates. Non-specific binding was 14.3 2.390 of 70 total binding. - with Specific binding | ZNI ORSRH; |ZNI СО5-21680 and |"25I|- MESA was determined as the difference between total h binding and non-specific binding. The percent inhibition of the compounds was calculated as a ratio to -» total binding. Competitive binding data were analyzed by constructing an iterative curve for a single-site model, and K values were calculated based on ISvo values (Spepo 5 Rgizoi, Viospet.RPpagtasoi. 22, 3099-3109, 1973) using the software SgarpRai Rgigt 2.01. -and Results 7 The main function of some receptors the cell surface is the recognition of the corresponding ligands. Accordingly, we determined the ligand binding affinities to confirm the functional integrity of the adenosine receptor subtype 7 expressed in yeast. Crude membranes obtained from 50 Zasspagotusevs sehemizvide transformed with the human adenosine receptor subtype 1 construct , showed specific binding when saturated with | "NIORSRH, where Ko was 4.0-0.19nM. Kr and Vp values were calculated from the saturation isotherm, and Scatchard transformation of the data indicated one class of binding sites. Estimates of the density of adenosine binding sites in yeast membrane preparations were made, which were 716.8-43.4 fmol/mg membrane protein. 29 Pharmacological characteristics of receptor subtypes for recombinant yeast cells transformed

HF) human adenosine receptor of the AT subtype was studied using selective adenosine ligands of several subtypes (HAS, ORSRH, Со5-15943, СО5-046142, СО5-046123, MESA, (K)-RIA, IV-MESA and alloxazin). They competed with -NIORSRH with an affinity of the expected order. The displacement curve plotted for these compounds showed a typical steepness for all ligands, and the data for each of these ligands could be modeled by a single-site fit. The apparent dissociation constants estimated for each compound from these curves (Table 5) are in agreement with the value published for the receptor obtained from other sources. because Ki values for yeast cell membranes transformed with human adenosine receptor subtype A! -dA-

MESA 179.6 (level

IV-MESA 606.5 vedy o soe-046123 vv

Tables 6-12 illustrate the efficacy and activity profiles of the deazapurine structures of the present invention. IN

Tables 13 and 14 illustrate the selectivity that can be achieved for human adenosine receptor sites by modulating the functional groups in the deazapurine structure. Table 14 also 7/5 illustrates the discovery that the compounds discussed here have subnanomolar activity and higher selectivity with respect to to the Ag receptor in comparison with the compounds presented in

Tables 13.

Table 6

Activity of series СО5-046142: Effect of M-6 substitute

MNE m. t a

M Y U-Me ek u

In our country

Binding Yeast

Ki code (nmM) 1050 (nm)

Wed -046142 139 972

SYU5-062365 1423 510,000 (ze)

Cr

СО5-0659533 ono 35 x10.006 in and -

SB8-069534 on 196.6 2425

AT-"

СО5056176 food GE 510,000 50000 0 nya 8 s СО5-056175 n 510000 510000 . nOUs?

СО5-062352 2975 510000 -о-. -i ime) -i (95) 4) ime) bo b5 sr 090909 on | 2 and NO c) sB8-090910 ды 1935 Y o

SRB-090913 I 415 yu Zh

Nick)

He

So5-092474 648 and "E

Tiaph (5.5)

Wed 092475 67260 m o

Jnas

Tigape (KK) soe-091175 NO, 321 08; From 7 o'clock (a

Sat 0635 "a 8169 25176 o (4) co

СО5-000914 ono mZ a, m tyas ch

Table 7 h-

Activity of series СО5-046142: Influence of Со-deputy 7 «он

WE Me « we -

Fo Adm and: her . "» n i - | Binding Yeast,

Code Ki (VM) IS50 (nm) yu Sre-060532 6045 510000

Mo" - S y o СО8-090595 157.1 7634 y v o I ko bo b5 -4v-

SB 065564 ha 1985 21875

AND"

SB5-090896 4436 510000

WITH

СО5-090903 b 2970 с 5

SOB-090890 ha Ete 1947 5

СО8-090915 195

IS

СБ5-090912 from 828

E and r

СО5.090177 Э 287 or sch shiya o

Table 8

Activity of the СО5-046142 series: The influence of the pyrrole ring substituent mon (Ce) 30 in vm Ge) in vv. what

M

VE

: с 35 че пн ни а о ' Connection Yeast 1 zapnia 1C50 1 Ki(vM) | (pit)

Code « sB8- Me Me Me 133 510000 z s 078187 "z SV. N Me i 123 1483 " 090905 s sB5- i n 15 75 090921 Sg ' -i

Wed Me Me 1210 510006 de 2-0 ooo ; -And (about)

SOV- x t Me Me |86341 4; sk -U Me Me 145 056091 me so Me Me TVBI in what

HF) i soB- x F Me Me 1353 z oi " in Table 9 b5 -D7-

MNVoome village

IN

M M

OO

Me putu niny sli " I will connect Drizhd to -YAE (nm)

СОВ- 18631 ше сБ5- di Bi? 056091 from

СО5- zones | 8A51 056089

Sat. 353 she

Table 10

Activity of series СО5-046123: Influence of Meo-deputy "MNE

Me with 7

M o

NN app nny and

Binding Yeast so

Ki code (nm) IC50 (nm) p

S5-067146 n 544 p

AND

Zo yy - yy

Wed-046123 n 828 . tr « o 70 C8-062357 МН 267 195.7 no) s va u . "» sBeovo35 535 to t - o -I name) -I (95) 4) name) 60 b5

СО5-062356 147.5 13972 pcs

Go

SB5-067325 I at 157 2918 er

СИ5-062392 at 692.5 510000 а ше

SIP5-062394 in in, | 475.3 2 T0000 et y

SOe-0b6956 2335 3462 t "y

СО8-067038 at 21 30095 hai

СО5-062359 73 3712 mouth

СО5-062360 ono 171 5054 sch roya o

Activity of the СО5B-046123 series: Effect of Mo-substitute 6

MNC Me

M x (Se) | Me so I

M and in se

IYUZYUZ0Z5 i u

Binding Yeast

In Ki (nm) 1C50 (nm) neo Ya

С5-069535 хи 53.9 551.0 - со п СО5-090894 мно днме | 103 101.3 o

SYUB-062301 many studies 74 33 - i

S5-090904 n 65 58.7 ko M. sn, ok - i

СО8-090906 I 1054 hud

M lo.

Me 23 4) her ko bo b5

C8-090908 « n 274 1624

And N

That

S5-090967 SHI from 1265 12970 te 62) 710 C0O85-095450 M.

MNAs 5 so5-095451 MNAS 173 5 i

EC

SELF: 713 wounds and to her

Table 12 "Retro-amide" analogues of СО5-046123

MNK me sch r» o

Me hk su her

IN

(Se) and Fo

Binding Yeast

The code is

СО5 065557 at 15 1854 p

Kv, h-

Со5-050591 at 74 lodge km

SB5- 062373 "А, 953 33450 "

GU - s СО5-090893 o 5 40406 p um "» y

SRV-062371 10600 210000 o - and o SO5-062372. o 127 I -I circles 20 СО5-065566 50.8 4025 4) and

СО8-065565 Sarchan 485 7015 o

HF) Table 13 7 Profile of selective antagonists of adenosine 60 b5 -BO0-

d

NK on

M. H

Me 2 ah M

H Binding of Ki (NM) and pe

СО8-046123. "victim 3139

Me

UA on spe-090908 | 218 507 |v46 14298 and new

СО5-090894 at 22 1756201 |43 ! oh ze une

СО5-090891 114 pz 01206 446 оон

КИ 139- сРе-046142 з09 19337 1380 |215 с "ОН ге) sp9-0908901 cho t09 |305 он kА and со8-0909052 164 7663 1683. |77

Go)

ON

(c) - со8-090909 2941. 1906 |1m30 |z4 s on

NU | what-

SOV-90910 IZ) 180 | from 6 10

SHE

M. Me "

M - (9) . ;from dream, in kne so5-121180 n 255 tv" 125 25 -y in) -I (snah, "me

І with sre-2yB and 531 9В1 t |53 y o - (snak., "nie

In Sat 070 00 sova» N mu |ze65 375 | "62 4) o z sna i sor5-123262 H 3095 6535 515 24 55 . tire o ON M NI

Phi so

СО5-062391 (Snu MNE 87 204 30 002 ko !

H bo b5 -5B1-

and about sleep con so8-121181 and 75,000 |720,000 13.00 1507 b

Kisnak, AH

СО5-121268 and 333 ti0,o0yu | 710,000 | 97 (in Ry

OD: сО5-121272 of your | 710,000 | 720000 1,369 mon

I 6305 | 2307k so5-096370" 305 1564 | 926 630476

ON

СО85-1137605 cho 18 206 80? 2 tons 720 Z; Wed-11666514 531 530 419

ОН соБ-131921 чо 111031 вай с: (o) 12-thienyl,-2-yl; 2С5-Н; Water-soluble; "B»5 and Kye mean hydrogen; "P3 means 3-fluorophenyl; "B43 means

C-chlorophenyl; "Vaz stands for 4-pyridyl; 895 activity at 10 µM.

Table 14

Profile of selective antagonists of adenosine ise) (d) -Y I so

Mn se wa i-

MTM hk n

ХЕ Ко code | Binding data К; 1 2 ID in i -o (nm) - from the soviet sovlyazaev "O-RI(r)E and so5azevi ORNI Me vot (03 50 sh ridinoon Me so. srelazeeee MERA ko -I Introduction as a link

All patents, published patent applications, and other works described herein are incorporated herein by reference. (95) 50 Equivalents

Ф It is obvious to any person skilled in the art, or any person skilled in the art, through only routine experimentation, can ascertain that there are many options equivalent to the specific implementation options of this invention described in this application. Such equivalents are included in the scope of the following claims. at

Claims (1)

The formula of the invention is 1. M-6-substituted 7-deazapurine of formula (I) bo b5 You Tom and ru UM y: VOM yu vo 9 where: each of K. and Ko independently represents a hydrogen atom, substituted (C.4-Cd)alkyl with a straight chain, substituted (C.3-Cd)alkyl with a branched chain, substituted (C., -C.-O)cycloalkyl, substituted cyclopropyl or substituted or unsubstituted aryl group; where only one of K. and Co. can be hydrogen; where, when the substituted straight-chain (C.14-C.sub.20)alkyl is -CH.sub.5-phenyl or -CH.-CH.sub.2-phenyl, then the specified phenyl is substituted; or Ku and B» together form a substituted or unsubstituted heterocyclic ring; Ez is a substituted or unsubstituted aryl group, Ku; represents a hydrogen atom, an unsubstituted alkyl, or a substituted or unsubstituted aryl group; each of Qui Co independently represents a halogen atom, a hydrogen atom, or a substituted or unsubstituted alkyl, aryl, or alkylaryl group, or K,. and K5 or Kb and K; together form a substituted or unsubstituted heterocyclic or carbocyclic ring, or a pharmaceutically acceptable salt thereof. 2. The compound according to claim 1 of the formula (1): s Tov i o Tom a F zjan cz Z so i chi - V sch de: each of K. and Ko independently represents a hydrogen atom, substituted (S.4-Szd) alkyl with a straight chain, substituted - (C3-Cd)alkyl with a branched chain, substituted (C.,-C.-O)cycloalkyl, substituted cyclopropyl or a substituted or unsubstituted aryl group; where only one of K. and Co. can be hydrogen; " du de, when substituted (C.14-C20)alkyl with a straight chain is -CH-C5-phenyl or -CH.-CH2-phenyl, then the specified phenyl is substituted; or with Ku and B" together form a substituted or unsubstituted heterocyclic ring; :z» Ez represents a substituted or unsubstituted aryl group, Ku represents a hydrogen atom, unsubstituted alkyl or a substituted or unsubstituted aryl group; each of Kb and Ke independently represents a halogen atom, a hydrogen atom or a substituted or unsubstituted alkyl, aryl or -1 15 alkylaryl group, or a pharmaceutically acceptable salt thereof. ko 3. The compound according to claim 2, where: -1 Ku represents hydrogen, | And | | And Co is independently substituted (C 4-C 20 )alkyl with a straight chain, substituted (C 3-C 20 )alkyl. with (95) 20 branched chain, substituted (C.-C.O)cycloalkyl, F or Ku and Co together form a substituted or unsubstituted heterocyclic ring, Ez is a substituted or unsubstituted aryl, Ku is a hydrogen atom, and each of Kb and Ke is independently hydrogen or alkyl, or a pharmaceutically acceptable salt thereof. HF) 4. The compound according to claim 3, where Kz is a substituted (C./-C.o)cycloalkyl. 7 5. Compound according to claim 4, where Ku and K are hydrogen; Kz is unsubstituted or substituted phenyl, and each of Kb and Ko is alkyl. 6. The compound according to claim 4 or claim 5, where K5 is monohydroxycyclopentyl. bo 7. The compound according to claim 4 or claim 5, where Co is monohydroxycyclohexyl. 8. The compound according to clause C, where K and Co together form a substituted or unsubstituted heterocyclic ring. 9. The compound according to claim 8, where the specified heterocyclic ring is substituted with amine or acetamide. 10. The compound according to claim 3, where Co represents -А-МНО(-О)В, where A represents an unsubstituted C 4 -C alkyl, and B represents a substituted or unsubstituted C 1 -alkyl. bo 11. The compound according to claim 10, where K. and K. represent hydrogen, Kz represents unsubstituted or substituted phenyl, and each of Kb and Ko represents alkyl. 12. The compound according to claim 3, where Kz represents a substituted or unsubstituted phenyl. 13. The compound according to claim 12, where each of Kb and Co is alkyl. 14. The compound according to claim 13, where Kz is unsubstituted phenyl. 15. The compound according to claim 13, where Kz is a substituted phenyl. 16. The compound according to claim 15, where Kz is phenyl substituted with at least one substituent. 17. The compound according to item C, where Kz represents a substituted or unsubstituted aryl, where aryl represents a heteroaryl group. 70 18. The compound according to claim 17, where each of Kb and Co is alkyl. 19. The compound according to claim 3, where each of K5b and Co represents hydrogen. 20. The compound according to item C, where each of Kb and K5 is methyl. 21. The compound according to claim 3, where the specified compound is selected from the group including: 4-(cis-3-hydroxycyclopentyl)amino-5,6-dimethyl-2-phenyl-7H-pyrrolo|2,3 d|pyrimidine, salt trifluoroacetic acid 4-(cis-3-(2-aminoacetoxy)-cyclopentyl)amino-5,6-dimethyl-2-phenyl-7H-pyrrolo|2,3 a|Ipyrimidine, 4-(3-acetamido)piperidinyl,- 5,6-dimethyl-2-phenyl-7H-pyrrolo-I(2,3 d|pyrimidine, 4-(2-M'-methylureidopropyl)amino-5,6-dimethyl-2-phenyl-7H-pyrrolo|2 ,3 a|Ipyrimidine, 4-(2-acetamidobutyl)amino-5,6-dimethyl-2-phenyl-7H-pyrrolo-(2,3 a|pyrimidine, 4-(2-M'-methylureidobutyl)amino-5 ,6-dimethyl-2-phenyl-7H-pyrrolo|2,3 4a|pyrimidine, 4-(2-aminocyclopropylacetamidoethyl)amino-2-phenyl-7H-pyrrolo(2,3 d|pyrimidine, 4-(trans-4 -hydroxycyclohexyl)amino-2-(3-chlorophenyl)-7H-pyrrolo|2,3 a|pyrimidine, 4-(trans-4-hydroxycyclohexyl)amino-2-(3-fluorophenyl)-7H-pyrroloY(2,3 d|pyrimidine, 4-(trans-4-hydroxycyclohexyl)amino-2-(4-pyridyl)-7H-pyrrolo|2,3 4a|pyrimidine. 22. M-6-substituted 7-deazapurine of the formula ІІІ, where: i) в М М- х оду иш зоря (O- осо С, in М и и и- В where сх X represents M or SK, - each of K. and Ko independently represents hydrogen or substituted or unsubstituted alkoxy, aminoalkyl, alkyl, aryl or alkylaryl, where only one of K. and Ko can be hydrogen, or "Ku and B" together form a substituted or unsubstituted heterocyclic ring; - 70 Ez represents a substituted or unsubstituted arylalkyl, aryl or heteroaryl; c KE. represents hydrogen; Iz» Y represents hydrogen, substituted or unsubstituted alkyl, or K y and Y taken together form a substituted or unsubstituted heterocyclic or carbocyclic ring; represents hydrogen, substituted or unsubstituted alkyl or halogen, O represents CH", O, C or MK), where K; represents hydrogen or substituted or unsubstituted C-Salkyl and and M/ represents unsubstituted or substituted alkyl, cycloalkyl, alkynyl, aryl, arylalkyl, biaryl, heteroaryl, or substituted carbonyl, substituted thiocarbonyl, or substituted sulfonyl. 23. The compound according to claim 22, where OO is CH, О, 5 or МН. She 24. The compound according to claim 23, where K is hydrogen, Y! represents hydrogen or methyl, and K z represents co 20 unsubstituted or substituted aryl. 25. The compound according to claim 24, where MU is substituted or unsubstituted alkyl, cycloalkyl, alkynyl or arylalkyl. that 26. The compound according to claim 25, where MU is alkynyl. 27. The compound according to claim 26, where MU is substituted by one or more substituents selected from the group consisting of halogen, hydroxy, substituted or unsubstituted alkyl, cycloalkyl, aryl or arylalkyl or MNK.o, where Co 25 is hydrogen or substituted, or unsubstituted alkyl, cycloalkyl, aryl or arylalkyl. GF) 28. The compound according to claim 25, where MU is substituted or unsubstituted cyclopentyl. 29. The compound according to claim 24, where M/ represents -(СН 2)4-С(-О)У or - (СНо)а-С(-5)У, where a is equal to 0, 1, 2 or 3 , U represents aryl, alkyl, arylalkyl, cycloalkyl, heteroaryl, MNK 44K.2, or, provided that when C) represents MH, then OK", and where each of K.4, K4" and Kz independently represents is hydrogen or unsubstituted or substituted alkyl, 60 aryl, arylalkyl or cycloalkyl. 30. The compound according to claim 23, where Kz is selected from the group consisting of substituted and unsubstituted phenyl, pyridyl, pyrimidyl, pyridazinyl, pyrazinyl, pyrrolyl, triazolyl, thiazolyl, oxazolyl, oxadiazolyl, pyrazolyl, furanyl, methylenedioxyphenyl and thiophenyl. 31. The compound according to claim 30, where Kz is unsubstituted phenyl. bo 32. The compound according to claim 30, where Kz is phenyl substituted with at least one substituent. -B4- 33. The compound according to claim 2 or claim 23, where K. and Co" together represent 79 and M where n is equal to 1 or 2, and where the ring may be optionally substituted by one or more groups: hydroxyl, amino, aminocarbonyl, thiol , carboxyl, halogen, СНоОН, СНО.МНО(-О)alkyl or SN.MNOT(-O)MNalkylom. 34. The compound according to claim 33, where n is 1 or 2, and the indicated ring is substituted by -MNO(-O)alkyl. 35. The compound according to claim 23, where K/ represents hydrogen, Ko represents substituted or unsubstituted C.4-Svalkyl, Kz represents substituted or unsubstituted phenyl, K/; represents hydrogen, and represents hydrogen either substituted or unsubstituted S.--Svalkil, O represents O, 5 or MK 7, where K; represents hydrogen or substituted or unsubstituted S.-Svalkyl, and M/ represents substituted or unsubstituted aryl. 36. The compound according to claim 35, selected from the group including 4-(2-acetylaminoethyl)amino-b6-phenoxymethyl-2-phenyl-7H-pyrrolo|2,3 a|pyrimidine, 4-(2-acetylaminoethyl)amino- 6-"4-fluorophenoxy)methyl-2-phenyl-7H-pyrrolo|2,3 d|pyrimidine, 4-(2-acetylaminoethyl)amino-6-"4-chlorophenoxy)methyl-2-phenyl-7H-pyrrolo| 2,3 d|pyrimidine, 4-(2-acetylaminoethyl)amino-6-(4-methoxyphenoxy)methyl-2-phenyl-7H-pyrrolo (2,3 a|Ipyrimidine, 4-(2-acetylaminoethyl)amino-6 -(2-pyridyloxy)methyl-2-phenyl-7H-pyrrolo|2,3-4|pyrimidine, 4-(2-acetylaminoethyl)amino-6-(M-phenylamino)methyl-2-phenyl-7H-pyrrolo| 2,3 d|Ipyrimidine, 4-(2-acetylaminoethyl)amino-6-"M-methyl-M-phenylamino)methyl-2-phenyl-7H-pyrrolo|2,3 d|pyrimidine, 4-(2-M '-methylureidoethyl)amino-β6-phenoxymethyl-2-phenyl-7H-pyrrolo|2,3 d|pyrimidine. 37. Use of a compound according to any one of claims 2, 3, 22 or 23 for the preparation of a medicinal product, which is used to treat a disease or condition associated with increased levels of adenosine in a patient, where the disease or condition is a disease of the central nervous system, kidney disease, or inflammatory disease, allergic disease, gastrointestinal disease, eye disease, or respiratory disease. 38. A compound according to any one of claims 2, 3, 5, 11, 22 or 23 for use in a method of inhibiting the activity (Ce) of an adenosine receptor in a cell. co 39. The compound according to claim 38, where the indicated compound is selected from the group consisting of: 4-(2-acetylaminoethyl)amino-b6-phenoxymethyl-2-phenyl-7H-pyrrolo|2,3 a|pyrimidine, - 4-( 2-acetylaminoethyl)amino-6-"4-fluorophenoxy)methyl-2-phenyl-7H-pyrrolo (2,3 Fpyrimidine, sc 4-(2-acetylaminoethyl)amino-6-(4-chlorophenoxy)methyl-2-phenyl -7H-pyrrolo|2,3 d|pyrimidine, 4-(2-acetylaminoethyl)amino-6-(4-methoxyphenoxy)methyl-2-phenyl-7H-pyrrolo|2,3 a|pyrimidine, - 4-(2 -acetylaminoethyl)amino-6-(2-pyridyloxy)methyl-2-phenyl-7H-pyrrolo|2,3 a|pyrimidine, 4-(2-acetylaminoethyl)amino-6-(M-phenylamino)methyl-2-phenyl -7H-pyrrolo|2,3 d|pyrimidine, 4-(2-acetylaminoethyl)amino-6-«"M-methyl-M-phenylamino)methyl-2-phenyl-7H-pyrrolo|2,3 4|pyrimidine, "4-(2-M'-methylureidoethyl)amino-β6-phenoxymethyl-2-phenyl-7H-pyrrolo|2,3 d|pyrimidine. 40. The compound according to claim 38, where the specified compound is selected from the group consisting of: t c 4-(cis-3-hydroxycyclopentyl)amino-5,6-dimethyl-2-phenyl-7H-pyrrolo|2,3 a| pyrimidine, h salt of trifluoroacetic acid 4-(cis-3-(2-aminoacetoxy)- » cyclopentyl)amino-5,6-dimethyl-2-phenyl-7H-pyrrolo|2,3 d|Ipyrimidine, 4-(3- acetamido)piperidinyl,-5,6-dimethyl-2-phenyl-7H-pyrrolo-I(2,3 d4|pyrimidine, 4-(2-M'-methylureidopropyl)amino-5,6-dimethyl-2-phenyl -7H-pyrrolo|2,3 d|pyrimidine, - 4-(2-acetamidobutyl)amino-5,6-dimethyl-2-phenyl-7H-pyrrolo-(2,3 a|Ipyrimidine, with 4- (2- M'-methylureidobutyl)amino-5,6-dimethyl-2-phenyl-7H-pyrrolo|2,3 d|pyrimidine, 4-(2-aminocyclopropylacetamidoethyl)amino-2-phenyl-7H-pyrrolo|2,3 a |pyrimidine, - 4-(trans-4-hydroxycyclohexyl)amino-2-(3-chlorophenyl)-7H-pyrrolo|2,3 a|pyrimidine, about 20 4-(trans-4-hydroxycyclohexyl)amino-2-( 3-fluorophenyl)-7H-pyrrolo|2,3 d|pyrimidine and 4-(trans-4-hydroxycyclohexyl)amino-2-(4-pyridyl)-7H-pyrroloY(2,3 d|pyrimidine. 42) 41. Use of the compound according to any of claims 2, 3, 5, 11, 22, and 23 for the preparation of a medicament for the treatment of central nervous system disease, cardiovascular disease, kidney disease, inflammatory disease, allergic disease, gastrointestinal disease, or respiratory disease in animals. Gee! 42. A pharmaceutical composition comprising a therapeutically effective amount of a compound according to any one of claims 2, 3, 5, 11, 22 and 23 and a pharmaceutically acceptable carrier. where 43. Pharmaceutical composition according to claim 42, where the specified compound is selected from the group consisting of 4-(2-acetylaminoethyl)amino-b6-phenoxymethyl-2-phenyl-7H-pyrrolo|2,3 a|pyrimidine, 60 4- (2-acetylaminoethyl)amino-6-(4-fluorophenoxy)methyl-2-phenyl-7H-pyrrolo|2,3 a|pyrimidine, 4-(2-acetylaminoethyl)amino-6-(4-chlorophenoxy)methyl-2 -phenyl-7H-pyrrolo|2,3 d|pyrimidine, 4-(2-acetylaminoethyl)amino-6-(4-methoxyphenoxy)methyl-2-phenyl-7H-pyrrolo|2,3 4|pyrimidine, 4-( 2-acetylaminoethyl)amino-6-(2-pyridyloxy)methyl-2-phenyl-7H-pyrrolo|2,3 a|pyrimidine, 4-(-2-acetylaminoethyl)amino-6-(M-phenylamino)methyl-2 -phenyl-7H-pyrrolo|2,3 Z|pyrimidine, 65 4-(2-acetylaminoethyl)amino-6-«"M-methyl-M-phenylamino)methyl-2-phenyl-7H-pyrrolo|2,3 4 pyrimidine, 4-(2-M'-methylureidoethyl)amino-6-phenoxymethyl-2-phenyl-7H-pyrrolo(2,3 da|pyrimidine. 44. Pharmaceutical composition according to claim 42, where the specified compound is selected from the group including: 4-(cis-3-hydroxycyclopentyl)amino-5,6-dimethyl-2-phenyl-7H-pyrrolo|2,3 a|pyrimidine, trifluoroacetic acid salt 4-(cis-3-(2-aminoacetoxy)-cyclopentyl)amino-5,6-dimethyl-2-phenyl-7H-pyrrolo|2,3 d|Ipyrimidine, 4-(3-acetamido)piperidinyl, -5,6-dimethyl-2-phenyl-7H-pyrrolo-I(2,3 d|pyrimidine, 4-(2-M'-methylureidopropyl)amino-5,6-dimethyl-2-phenyl-7H-pyrrolo| 2,3 a|Ipyrimidine, 4-(2-acetamidobutyl)amino-5,6-dimethyl-2-phenyl-7H-pyrrolo-(2,3 a|pyrimidine, 4-(2-M'-methylureidobutyl)amino- 5,6-dimethyl-2-phenyl-7H-pyrrolo|2,3 4a|pyrimidine, 70 4-(2-aminocyclopropylacetamidoethyl)amino-2-phenyl-7H-pyrrolo(2,3 d|pyrimidine, 4-(trans -4-hydroxycyclohexyl)amino-2-(3-chlorophenyl)-7H-pyrrolo|2,3 a|pyrimidine, 4-(trans-4-hydroxycyclohexyl)amino-2-(3-fluorophenyl)-7H-pyrrolo|2 ,3 d|pyrimidine and 4-(trans-4-hydroxycyclohexyl)amino-2-(4-pyridyl)-7H-pyrrolo|2,3 4a|pyrimidine. 45. The pharmaceutical composition according to claim 42, wherein said therapeutically effective amount is effective for /5 Treatment of respiratory disease or gastrointestinal disease. 46. Pharmaceutical composition according to claim 45, where the specified gastrointestinal disease is diarrhea. 47. Pharmaceutical composition according to claim 45, where the indicated respiratory disease is asthma, allergic rhinitis or chronic obstructive pulmonary disease. 48. Pharmaceutical composition according to claim 42, where the indicated pharmaceutical composition is an ophthalmic preparation. 49. Pharmaceutical composition according to claim 48, where the specified drug is a drug for injections into the peri-ocular, retrobulbar or intraocular area. 50. Pharmaceutical composition according to claim 48, where the specified drug is a drug for systemic administration. with 51. Pharmaceutical composition according to claim 48, where the specified drug is a surgical solution for washing. and) 57. A packaged pharmaceutical composition for the treatment of an adenosine-mediated disease or disorder, wherein the disease is a central nervous system disease, cardiovascular disease, renal disease, inflammatory disease, allergic disease, gastrointestinal disease, or "o zo respiratory disease" in a mammal which contains: a container containing a therapeutically effective amount of at least one compound according to claims 2, 3, 5, 11, 22 or 23; and i- instructions for the use of said compound for the treatment of a disease or disorder mediated by adenosine in a mammal. with 53. Method of obtaining cha ICC ! and S i : MM t s Z i: . n "» t. which includes the stages: a) interaction of 47 МО V, m ЖЖ M mon - syu 70 РФ and (9) va o with obtaining ko Me , nok in Н R where R is a protective group that is removed; bo 5) treatment of the product of stage (a) under conditions of cyclization to obtain -58c- ON V, ' M xx Av | H in MK H c) treatment of the product of stage (B) under acceptable conditions with the production of 70 С 6 M Kon V MK n and a) treatment of the chlorinated product of stage (c) with an amine with the production ICC in ' 6 Mch rya in MK H s where o each of K. and Co independently represents a hydrogen atom or a substituted (C 4-Czo)alkyl with a straight chain, a substituted (C53-Si)alkyl with a branched chain, substituted (C) -C.00)cycloalkyl, substituted cyclopropyl or substituted or unsubstituted aryl group; where only one of K. and K" can be hydrogen; (se) where, when the substituted (C.14-C.sub.20)alkyl with a straight chain is -CH.sub.5-phenyl or -CH.-S.sub.2-phenyl, then the specified phenyl is substituted; or Ku and KE" together form a substituted or unsubstituted heterocyclic ring; - Ez represents a substituted or unsubstituted aryl group and sch Ko; represents a halogen atom, a hydrogen atom, or a substituted or unsubstituted alkyl, aryl, or C5 alkylaryl group. - 54. The compound according to claim 7, having the structure no, ХХ - с and СН. and? sh- | х сн чв Bi M Z -i N ime) - 55. The compound according to claim 7, which has a structure of 50 NO. I "c 2, nya N yet F. My name) M M n 60 56. The compound according to claim 7, having the structure b5 NMye. 0- he is more than E 57. The compound according to claim 7, which has the structure zhe pe Z re МНС с 29 58. The compound according to claim 7, which has the structure (о) (Се) то И: И со p MM - - Ме и - 59. The compound according to claim 11, which has the structure ik « rshi with NO. z i ni o SN z To sn o 50 vn. M Z 42) 60. The compound according to claim 11, which has the structure Ф) име) 60 b5 NOM WE. U nya NU te xx na | M SNU M n 61. The compound according to paragraph Z, which has the structure и s Mn o Bo I M sh:y o tiy ih - сн сх з M Z N i - 62. The compound according to item C, which has the structure " SN. . - with НМ 4; - - HO ny -I ime) -I M o 50 SN o tscha their re | 7 M M she F) by 63. p. The compound according to item C, having the structure 60 b5 We are MN. 710 u nya SV i p -h SN M M Z Y 64. A compound according to claim 9 having a structure Nn. M. "SN she s (o) o SN. M | Kh re: moz Te M N so cha 65. The compound according to claim 64, which has the structure с н' иту What about « сн З - - с М | their most common SN and M M -I from 66. The compound according to claim 64, having the structure NN. -and SN. ah IF (o) SN. 67. The compound of claim 2, wherein any substituent, if present, is halogen, hydroxyl, alkylcarbonyloxy, arylcarbonyloxy, alkoxycarbonyloxy, aryloxycarbonyloxy, carboxylate, alkylcarbonyl, alkoxycarbonyl, aminocarbonyl, alkylthiocarbonyl, alkyl, phosphate, phosphonato, phosphinato, cyano, amino, alkylamino, dialkylamino, arylamino, diarylamino, alkylarylamino, acylamino, alkylcarbonylamino, arylcarbonylamino, carbamoyl, ureido, amidino, imino, sulfhydryl, alkylthio, arylthio, thiocarboxylate , sulfonato, sulfamoyl, sulfonamido, nitro, trifluoromethyl, azido, heterocyclyl, alkylaryl, or an aromatic or heteroaromatic group, where the substituent in turn may be substituted by any of the above substituents. 68. The compound of claim 67, wherein any substituent, if present, is halogen, hydroxyl, alkylcarbonyloxy, arylcarbonyloxy, alkoxycarbonyloxy, carboxylate, alkylcarbonyl, alkoxycarbonyl, aminocarbonyl, alkylthiocarbonyl, alkyloxy, amino, alkylamino, dialkylamino, acylamino, alkylcarbonylamino. , arylcarbonylamino, carbamoyl, ureido, amidino, imino, nitro, heterocyclyl, alkylaryl, or an aromatic or heteroaromatic group, where the substituent in turn may be substituted with any of the above substituents. 69. The compound according to claim 68, where the substituent is halogen, hydroxyl, alkylcarbonyloxy, alkoxycarbonyloxy, 70 carboxylate, alkylcarbonyl, alkoxycarbonyl, aminocarbonyl, alkylthiocarbonyl, alkoxyl, amino, alkylamino, dialkylamino, acylamino, alkylcarbonylamino, arylcarbonylamino, carbamoyl, amidino, uridine imino, nitro, heterocyclyl or heteroaromatic group, where the substituent in its turn can be replaced by any of the above substituents. 70. The compound according to claim 67, where K and Co are substituted (C 4-C 20 ) alkyl with a straight chain or substituted 75. "C3-Sio)alkyl with a branched chain containing two substituents. Official bulletin "Industrial Property". Book 1 "Inventions, useful models, topographies of integrated microcircuits", 2005, M 4, 15.04.2005. State Department of Intellectual Property of the Ministry of Education and Science of Ukraine. s schi c) (Se) (ze) u s i - - a -i ko -i (95) 4) ko bo b5