RU2097384C1 - Derivatives of amino-7-(chr2r3)-3h,5h-pyrrolo-[3,2-d]-pyrimidine-4-one, methods of their synthesis and a method of selective inhibition of mammalian t-lymphocytes proliferation and no effecting on chr2r3-lymphocytes - Google Patents

Abstract

FIELD: organic chemistry. SUBSTANCE: invention proposes derivatives of 2-amino-7-(CHR₂R₃)-3H, 5H-pyrrolo-[3,2-d] -pyrimidine-4-one of the formula:

where: R₁ - H, NH₂; R₂ - saturated 5-membered heterocycle containing S or O as heteroatom, or 6-membered N-containing heterocycle that can be substituted; R₃ - H, CH₂, CH₂CO₂H, CH₂CONH₂, CO₂CH₃, CO₂H or CH₂CH₂OH,, or R₂ and R₃ together with CH form cyclohexyl- or cyclohexenyl-group. The synthesized compounds are inhibitors of purine-nucleoside phosphorylase. Invention proposes methods of their synthesis and a method of selective inhibition of mammalian T-lymphocytes proliferation but no effecting on B-lymphocytes. EFFECT: improved method of synthesis. 10 cl, 1 tbl

Description

The invention relates to new derivatives of 2-amino-7- (CH ₂ R ₂ R ₃ ) -3H, 5H-pyrrolo [3,2-d] pyrimidines having the properties of a selective T-lymphocyte inhibitor, methods for their preparation and a method for selective inhibition of proliferation Mammalian T lymphocytes and no effect on B lymphocytes.

 Purin nucleoside phosphorylase (PNP) is known to catalyze the phosphorolysis of purin nucleoside in a reversible reaction. In individuals lacking PNP, there is a weakened development of T-lymphocytes, leading to reduced intercellular immunity, and normal development of basophilic insulocyte, leading to normal humoral immunity. Accordingly, specific PNP inhibitors that selectively inhibit the development of T-lymphocytes without compromising humoral immunity can be potentially effective against diseases in which activated T-lymphocytes are pathogenic.

 The journal (Science, 1981, v. 214) describes 8-amino guanosines that have a selective effect on T cells without affecting B cells.

 New derivatives of pyrrolopyrimidinones, which are based on a bicyclic heterocyclic system, unexpectedly showed the property of selectively weakening the functions of mammalian T-lymphocytes without weakening the effect on humoral immunity, and due to these properties, which have been found and studied, they can be used to treat serious diseases such as autoimmune disorders, transplant rejection organs, as well as in the treatment of cancer.

где R₁ означает H или NH₂;
R₂ означает насыщенный пятичленный гетероцикл, содержащий в качестве гетероатома серу или кислород, или шестичленный гетероцикл, содержащий в качестве гетероатома N, которые могут быть необязательно замещены (C₁-C₄)алкокси, трифторметилом или галогеном; (C₆-C₁₀)циклоалкил, незамещенный или галоидзамещенный фенил;
R₃ означает H, (CH₂)_nCN, (CH₂)_nCOOH, CO₂CH₃, (CH₂)_nCONH₂, (CH₂)_nCO₂CH₃, (CH₂)_nOH, n 1-4, R₂ и R₃ вместе с CH-группой, к которой они присоединены, образуют циклогексил или циклогексенильную группу. Предпочтительны соединения, в которых R₂ и R₃ вместе с CH-группой, к которой они присоединены, образуют циклогексил или циклогексенил.The invention provides derivatives of 2-amino-7- (CHR ₂ R ₃ ) -3H, 5H-pyrrolo- [3,2-d] pyrimidin-4-one of formula (I)

where R ₁ means H or NH ₂ ;
R ₂ means a saturated five-membered heterocycle containing sulfur or oxygen as a heteroatom, or a six-membered heterocycle containing N as a heteroatom, which may optionally be substituted with (C ₁ -C ₄ ) alkoxy, trifluoromethyl or halogen; (C ₆ -C ₁₀ ) cycloalkyl, unsubstituted or halogenated phenyl;
R ₃ means H, (CH ₂ ) _n CN, (CH ₂ ) _n COOH, CO ₂ CH ₃ , (CH ₂ ) _n CONH ₂ , (CH ₂ ) _n CO ₂ CH ₃ , (CH ₂ ) _n OH, n 1-4, R ₂ and R ₃ together with the CH group to which they are attached form a cyclohexyl or cyclohexenyl group. Compounds in which R ₂ and R ₃ together with the CH group to which they are attached form cyclohexyl or cyclohexenyl are preferred.

Preferred are compounds where R _{1 is} NH ₂ group and compounds where R ₂ and R ₃ together with the adjacent CH group form a cyclohexyl or cyclohexenyl ring, or R ₂ phenyl, compounds where R ₃ -CH ₂ CN, CH ₂ COOH , CH ₂ CONH ₂ , or compounds where R _{2 is} optionally substituted with C ₁ -C ₄ alkoxy, halogen, trifluoromethyl, 3-piperidinyl, 2-tetrahydrofuranyl, 2-tetrahydrothienyl, 3-, 4-pyridinyl, cyclopentyl, cyclohexyl, cycloheptyl 2-adamantyl; or phenyl, 3-chlorophenyl, 2,3,5-trichlorophenyl.

Among the preferred compounds, mention may be made of 2-amino-7 (CH ₂ R) -3H, 5H-pyrololo [3,2-d] pyrimidin-4-ones (I ′), where R 2- or 3-tetrahydrothienyl, 2-, 3- or 4-piperidinyl, 2- or 3-tetrahydrofuranyl, 2-, 3- or 4-tetrahydrofuranyl. In particular, compounds of the invention selected from the group: 2-amino-7- (2-piperidinylmethyl) -3H, 5H-pyrrolo [3,2-d] pyrimidin-4-one (IB), 2-amino -7- (3-piperidinylmethyl) -3H, 5H-pyrrolo [3,2-d] pyrimidin-4-one (I'C), 2-amino-7- (4-piperidinylmethyl) -3H, 5H-pyrpolo [ 3,2-d] pyrimidin-4-one (I'D), 2-amino-7- (2-tetrahydrofuranylmethyl) -3H, 5H-pyrrolo [3,2-d] pyrimidin-4-one (I ' E), 2-amino-7- (3-tetrahydrofuranylmethyl) -3H, 5H-pyrrolo [3,2-d] pyrimidin-4-one (I'F), 2-amino-7- (2-tetrahydrothienylmethyl) - 3H, 5H-pyrrolo [3,2-d] pyrimidin-4-one (I'G), 2-amino-7- (3-tetrahydrothienylmethyl) -3H, 5H-pyrrolo [3,2-d] pyrimidin-4 -he ( I'H).

In these compounds, the heterocyclic radical R at position 7 may be substituted with one or two substituents selected from the group: halogen, (C ₁ -C ₄ ) alkoxy, trifluoromethyl.

 As halogen, chlorine or fluorine is preferred. As alkoxy, lower alkoxy, methoxy, ethoxy, propoxy and butoxy are preferred.

Among the compounds covered by the invention, mention may be made of 2-amino-7- (CH ₂ R) -3H, 5H-pyrrolo [3,2-d] pyrimidin-4-ones (II), in which R-pyridinyl, in particular compounds II, selected from the group, 2-amino-7- (3-pyridinylmethyl) -3H, 5H-pyrrolo [3,2-d] pyrimidin-4-one (IIA), 2-amino-7- (2-pyridinylmethyl ) -3H, 5H-pyrrolo [3,2-d] pyrimidin-4-one (IIB), or 2-amino-7- (4-pyridinylmethyl) -3H, 5H-pyrrolo [3,2-d] pyrimidine- 4-one (IIC) in which pyridinyl may have one or two substituents selected from the group consisting of halogen, (C ₁ -C ₄ ) alkoxy or trifluoromethyl. Chlorine or fluorine are preferred as halogen. As alkoxy, methoxy, ethoxy, propoxy and butoxy are preferred.

 Another group of preferred compounds are (III) 2-amino-7- (R) -3H, 5H-pyrrolo [3,2-d] pyrimidin-4-ones, in which R 1-, 2- or 3-cyclohexenyl or cyclohexyl . Among them are compounds III selected from the group: 2-amino-7- (1-cyclohexenyl) -3H, 5H-pyrrolo [3,2-d] pyrimidin-4-one (IIIA), 2-amino-7- (2-cyclohexenyl) -3H, 5H-pyrrolo [3,2-d] pyrimidin-4-one (IIIB), 2-amino-7- (3-cyclohexenyl) -3H, 5H-pyrrolo [3,2-d ] pyrimidin-4-one (IIIC) or 2-amino-7- (cyclohexyl) -3H, 5H-pyrrolo [3,2-d] pyrimidin-4-one (IIID).

The invention also provides 2-amino-7- (CH ₂ R) -3H, 5H-pyrrolo [3,2-d] pyrimidin-4-ones (IV), in which R are single ring cycloparaffins, such as, for example, cyclopentyl, cyclohexyl and cycloheptyl; multi-ring cycloparaffins, such as 1- and 2-adamantyl, 1-norbornanil, 2-exo-norbornanil, 2-endo-norbornanil, 1- and 2-bicyclo [2.2.2] octanyl, 1-, 2-, 3 -, 6- and 8-bicyclo [3.2.1] octanyl, and 1-, 2- and 3-bicyclo [3.3.1] nonanyl, and cycloolefins, such as, for example, 1- and 2-norbornenyl. Examples of this group of substances (IV) are 2-amino-7- (2-adamantylmethyl) -3H, 5H-pyrrolo [3,2-d] pyrimidin-4-one (IVA), 2-amino-7- (1- adamantylmethyl) -3H, 5H-pyrrolo [3,2-d] pyrimidin-4-one (IVB), 2-amino-7- (cyclopentylmethyl) -3H, 5H-pyrrolo [3,2-d] pyrimidin-4- he (IVC), 2-amino-7- (cyclohexylmethyl) -3H, 5H-pyrrolo [3,2-d] pyrimidin-4-one (IVD), 2-amino-7- (cycloheptylmethyl) -3H, 5H- pyrrolo [3,2-d] pyrimidin-4-one (IVE), 2-amino-7- (1-norbornanylmethyl) -3H, 5H-pyrrolo [3,2-d] pyrimidin-4-one (IVF), 2-amino-7- (2-exo-norbornanylmethyl) -3H, 5H-pyrrolo [3,2-d] pyrimidin-4-one (IVG), 2-amino-7- (2-endo-norbornanylmethyl) -3H , 5H-pyrrolo [3,2-d] pyrimidin-4-one (IVH), 2-amino -7- (1-norbornenylmethyl) -3H, 5H-pyrrolo [3,2-d] pyrimidin-4-one (IVI), 2-amino-7- (2-norbornenylmethyl) -3H, 5H-pyrrolo [3, 2-d] pyrimidin-4-one (IVJ), 2-amino-7- (1-bicyclo [2.2.2] octanylmethyl) -3H, 5H-pyrrolo [3,2-d] pyrimidin-4-one (IVK ), 2-amino-7- (1-bicyclo [3.2.1] octanylmethyl) -3H, 5H-pyrrolo [3,2-d] pyrimidin-4-one (IVL) and 2-amino-7- (1- bicyclo [3.3.1] nonanylmethyl) -3H, 5H-pyrrolo [3,2-d] pyrimidin-4-one (IVM), and 2-amino-7- (1-noradamantylmethyl) -3H, 5H-pyrrolo [3 , 2-d] pyrimidin-4-one (IVN).

Among the compounds obtained according to the invention, mention may be made of a group of derivatives of 2-amino-7- (CHR ₂ R ₃ ) -3H, 5H-pyrrolo [3,2-d] pyrimidin-4-one of the formula (I), where R ₁ , R ₂ are as defined above, and R ₃ is a group selected from H, CH ₂ CN, CH ₂ CO ₂ H, CH ₂ CONH ₂ , CO ₂ CH ₃ , CH ₂ CH ₂ OH, CH ₂ CO ₂ CH ₃ .

 Another object of the invention is a method for selectively inhibiting the proliferation of mammalian T-lymphocytes, which does not have an effect on B-lymphocytes, comprising administering to the mammal the above compounds of formula (I) in an effective amount, said compound inhibiting purine nucleoside phosphorylase and thereby the formation of T-lymphocytes .

 It is also possible for mammals to administer an effective amount of a compound of formula (I), which does not have a weakening effect on humoral immunity and has a selective weakening effect on T-lymphocytes, with a pharmaceutically acceptable diluent.

 A feature of the proposed method for the selective weakening of the function of mammalian T-lymphocytes without attenuating the effect on humoral immunity is that the administration to a mammal of a substance of formula (I) inhibits purine nucleoside phosphorylase and thereby the formation of T-lymphocytes.

 For enteral, such as oral or rectal, transdermal or parenteral administration to mammals, including humans, which are suitable for inhibiting the activity of purine nucleoside phosphorylase and for treating diseases susceptible to it, it is possible to administer an effective amount of a pharmacologically active substance of the invention alone or in combination with one or more pharmaceutically acceptable carriers.

 A preferred form of compositions used are tablets and gelatin capsules comprising the active ingredient together with: a) diluents, for example, lactose, glucose, sucrose, mannitol, sorbitol, cellulose and / or glycine; b) lubricants, for example, silicon dioxide, talc, stearic acid, its magnesium or calcium salt and / or polyethylene glycol; for tablets also with: c) binders, for example, magnesium aluminum silicate, starch paste, gelatin, tragacanth, methyl cellulose, sodium carboxymethyl cellulose and / or polyvinylpyrrolidone; if desired: d) disintegrants, for example, starches, agar-agar, alginic acid or its sodium salt, or effervescent mixtures; and / or e) absorbents, dyes, flavors and sweeteners. The injectable compositions are preferably aqueous isotonic solutions or suspensions, and suppositories are advantageously prepared from fat emulsions or suspensions. These compositions may be sterilized and / or contain adjuvants such as, for example, preservatives, stabilizing, wetting or emulsifying agents, dissolved promoters, salts for regulating the osmotic pressure and / or buffer solutions. In addition, they may also contain other therapeutically valuable substances. These compositions are prepared in accordance with standard methods of mixing, granulating or coating, respectively, and they usually contain about 0.1-75.0%, preferably about 1-50% of the active ingredient.

 For transdermal use, an effective amount of a substance of the invention with a carrier is used. Useful carriers include absorbable pharmacologically acceptable solvents in order to help pass into the body through the skin. It is essential that the transdermal devices can be in the form of a bandage comprising a supporting part, a reservoir containing a preferred substance with carriers, a barrier for delivering the substance to the skin of the body at a controlled and predetermined speed for an extended period of time, and means for attaching the device to the skin .

 The proposed method for the selective inhibition of the activity of purine nucleoside phosphorylase can be used in the treatment of diseases and media susceptible to them, for example, in autoimmune disorders, rejection during transplantation, psoriasis.

 The method is also suitable for inhibiting phosphorolysis and metabolic cleavage of antiviral or antitumor purinucleosides in mammals by administering a compound of formula (I) alone or with one or more pharmaceutically acceptable carriers. In particular, this relates to a method for inhibiting phosphorolysis and metabolic cleavage of known purinucleosides, for example, 2'-deoxyguanosine, 2 ', 3'-dideoxyinosine, 2', 3'-dideoxyguanosine or 2 ', 3'-dideoxyoxyadenosine.

 Thus, the invention also relates to a method for inhibiting phosphorolysis or metabolic cleavage of the antiviral or antitumor effect of 2'- or 3'-monodeoxypurin nucleosides or 2 ', 3'-dideoxypurin nucleosides in mammals, which comprises administering to mammals, if necessary, either alone or in combination with the indicated nucleoside of an effective amount of a substance of the invention inhibiting purine nucleoside phosphorylase, preferably in combination with one or more pharmaceutically acceptable carriers yami. More specifically, this relates to a method of activating or enhancing the effect of 2 ′, 3′-dideoxypurin nucleosides known in the art, for example 2 ′, 3′-dideoxyinosine, 2 ′, 3′-dideoxyguanosine or 2 ′, 3′-dideoxyadenosine for treatment retroviral infections, such as HIV retroviral infections (acquired immunodeficiency syndrome, AIDS). 2 ', 3'-dideoxypurine nucleosides are known as inhibitors of HIV retroviral infection and are metabolically cleaved by PNP, for example, as described in Biochemical Pharmacology, 22, 3797 (1987). They are administered in a pharmaceutically tolerated dose that is effective in inhibiting HIV retroviral infections. Preferably, the lowest possible effective dose is used.

 The pharmaceutically tolerated effective dosage of the active substance of the invention, which is administered, depends on the type of warm-blooded animals (mammals), body weight, age, individual condition and form of administration.

 The pharmaceutical composition may be oral, parenteral, suppository, or another form that delivers the substance into the blood stream of a mammal that needs to be treated. The oral form has from about 1 to about 150 mg of active substance for an adult (50-70 kg), which is mixed with pharmaceutically acceptable diluents, for example, lactose. In a regular capsule, 25 mg of the substance is mixed together with 192 mg of lactose, 80 mg of modified starch and 3 mg of magnesium stearate. It is also possible to use forms suitable for injection.

 The use of the compounds of the present invention is also possible with other therapeutic agents. The daily dosage for humans (weighing 50-70 kg is 1-50 mg / kg) inhibits the metabolic destruction of certain anti-cancer agents, for example, beta-2'-deoxy-6-thioguanazine and antiviral agents, for example, 2 ', 3'-dideoxyinosine , anti-AIDS remedies. These types of medicines are known to be unstable and destroyed, losing their effectiveness. The substances of the invention are able to reduce such instability, which increases the effectiveness of other chemotherapeutic agents.

где R₁ означает H, NH₂;
R₂ и R₃ означают незамещенный или галоидзамещенный фенил или циклогексил;
R₂ означает CH₂CN.The invention also relates to methods for preparing 2-amino-7- (CHR ₂ , R ₃ ) -3H, 5H-pyrrolo [3,2-d] pyrimidin-4-ones derivatives of the general formula

where R ₁ means H, NH ₂ ;
R ₂ and R _{3 are} unsubstituted or halogenated phenyl or cyclohexyl;
R ₂ means CH ₂ CN.

One method is intended for the preparation of compounds of the indicated formula, where R ₁ is NH ₂ , and the other is for compounds of the specified formula, where R ₁ is H.

где R₁ NH₂;
R₂ фенил, незамещенный или замещенный галоидом, или циклогексил;
R₃ CH₂CN, заключается в том, что осуществляют следующие последовательные стадии:
а) реакцию соответствующего R₂-замещенного альдегида с циануксусной кислотой в присутствии ацетата аммония с получением 3(R₂)-замещенного пентандинитрила;
б) реакцию полученного 3(R₂)-пентандинитрила с алкиловым эфиром муравьиной кислоты в присутствии основания, с получением 3(R₂)-2-формилпентандинитрила,
в) который взаимодействует с гидрохлоридом метилового эфира глицина в присутствии ацетата натрия или аммония с получением метил-N-[(3(R₂)-2,4-дициано)-2-бутенил]глицина;
г) полученный при этом N-[(3(R₂)-2,4-дициано)-2-бутенил]глицин взаимодействует с алкиловым эфиром хлоруксусной кислоты в присутствии 1,5-диазабицикло[4.3.0]-нон-5-еном или 1,8-диазабицикло[5.4.0]ундек-7-еном с получением метил-3-амино-4-(2-циано-1(R₂)-этил)-1-этил-1H-пирроло-1,2- дикарбоксилата;
д) получаемый метил-3-амино-4-(2-циано-1(R₂)-этил)-1-этил-1H-пирроло-1,2- дикарбоксилат обрабатывают основанием с получением метил-3-амино-4-(2-циано-1(R₂)-этил)-1H-пирроло-2-карбоксилата;
е) последний подвергают взаимодействию с бензоилизотиоцианатом с получением N-бензоил-N'-[4-(2-циано-1(R₂)-этил)-2-метоксикарбонил-1Н-пиррол-3-ил]тиомочевины;
ж) полученную N-бензоил-N'-[4-(2-циано-1(R₂)-этил)-2-метоксикарбонил-1H-пиррол-3-ил] тиомочевину алкилируют галоидным алкилом с получением N-бензоил-N'-[4-(2-циано-1(R₂)-этил)-2-метоксикарбонил-1H-пиррол-3-ил] -S-метилтиомочевины;
з) затем N-бензоил-N'-[4-(2-циано-1(R₂)-этил)-2-метоксикарбонил-1H-пиррол-3-ил] -S-метилтиомочевины обрабатывают метанольным или этанольным раствором аммиака с получением смеси 3(R₂)-3-[2-амино-4-оксо-3H,5H-пирроло[3,2-d] пиримидин-7-ил] пропаннитрила и 3(R₂)-3-[2-меркапто-4-оксо-3H,5H-пирроло[3,2-d]пиримидин-7-ил]пропаннитрила.The method of obtaining derivatives of 2-amino-7- (CHR ₂ R ₃ ) -3H, 5H-pyrrolo [3,2-d] pyrimidin-4-one (method A) of the formula (I ')

where R ₁ NH ₂ ;
R ₂ phenyl unsubstituted or substituted by halogen, or cyclohexyl;
R ₃ CH ₂ CN, lies in the fact that carry out the following sequential stages:
a) the reaction of the corresponding R ₂ -substituted aldehyde with cyanoacetic acid in the presence of ammonium acetate to obtain 3 (R ₂ ) -substituted pentanedinitrile;
b) reacting the obtained 3 (R ₂ ) -pentanedinitrile with formic acid alkyl ester in the presence of a base to obtain 3 (R ₂ ) -2-formylpentanedinitrile,
c) which interacts with glycine methyl ester hydrochloride in the presence of sodium or ammonium acetate to give methyl N - [(3 (R ₂ ) -2,4-dicyano) -2-butenyl] glycine;
d) the resulting N - [(3 (R ₂ ) -2,4-dicyano) -2-butenyl] glycine interacts with the chloroacetic acid alkyl ester in the presence of 1,5-diazabicyclo [4.3.0] non-5- ene or 1,8-diazabicyclo [5.4.0] undec-7-ene to give methyl 3-amino-4- (2-cyano-1 (R ₂ ) ethyl) -1-ethyl-1H-pyrrolo-1 2-dicarboxylate;
d) the resulting methyl-3-amino-4- (2-cyano-1 (R ₂ ) -ethyl) -1-ethyl-1H-pyrrolo-1,2-dicarboxylate is treated with a base to obtain methyl-3-amino-4- (2-cyano-1 (R ₂ ) -ethyl) -1H-pyrrolo-2-carboxylate;
e) the latter is reacted with benzoylisothiocyanate to give N-benzoyl-N ′ - [4- (2-cyano-1 (R ₂ ) ethyl) -2-methoxycarbonyl-1H-pyrrol-3-yl] thiourea;
g) the resulting N-benzoyl-N '- [4- (2-cyano-1 (R ₂ ) -ethyl) -2-methoxycarbonyl-1H-pyrrol-3-yl] thiourea is alkylated with alkyl halide to give N-benzoyl-N '- [4- (2-cyano-1 (R ₂ ) -ethyl) -2-methoxycarbonyl-1H-pyrrol-3-yl] -S-methylthiourea;
h) then N-benzoyl-N '- [4- (2-cyano-1 (R ₂ ) ethyl) -2-methoxycarbonyl-1H-pyrrol-3-yl] -S-methylthiourea is treated with a methanol or ethanol solution of ammonia with obtaining a mixture of 3 (R ₂ ) -3- [2-amino-4-oxo-3H, 5H-pyrrolo [3,2-d] pyrimidin-7-yl] propannitrile and 3 (R ₂ ) -3- [2- mercapto-4-oxo-3H, 5H-pyrrolo [3,2-d] pyrimidin-7-yl] propannitrile.

Preferably this method is used for compounds I ', where R ₂ phenyl.

где R₁ H;
R₂ фенил, незамещенный или замещенный галоидом, или циклогексил;
R₃ CH₂CN, заключается в том, что осуществляют последовательные стадии:
а) реакцию соответствующего циклического альдегида с циануксусной кислотой в присутствии ацетата аммония с получением 3(R₂)-замещенного пентандинитрила;
б) реакцию полученного 3(R₂)-пентандинитрила с алкиловым эфиром муравьиной кислоты в присутствии основания с получением 3(R₂)-2-формилпентандинитрила;
в) последний подвергают взаимодействию с гидрохлоридом метилового эфира глицина в присутствии ацетата натрия или аммония с получением метил-N-[(3(R₂)-2,4-дициано)-2-бутенил]глицина;
г) полученный метил-N-[(3(R₂)-2,4-дициано)-2-бутенил]глицин подвергают взаимодействию с алкиловым эфиром хлоруксусной кислоты в присутствии 1,5-диазабицикло[4.3.0] нон-5-ена или 1,8-диазабицикло[5.4.0]ундек-7-ена с получением метил-3-амино-4-(2-циано-1(R₂)-этил)-1-этил-1H-пиррол-1,2- дикарбоксилата;
д) метил-3-амино-4-(2-циано-1(R₂)-этил)-1-этил-1H-пиррол-1,2-дикарбоксилат обрабатывают основанием с получением метил-3-амино-4-(2-циано-1(R₂)-этил)-1H-пирроло-2-карбоксилата;
е) затем метил-3-амино-4-(2-циано-1(R₂)-этил)-1H-пирроло-2-карбоксилат подвергают взаимодействию с диметилацеталем диметилформамида с получением метил-4-(2-циано-1(R₂)-этил)-3-[N-(диметиламино-метилен)амино] -1H-пирроло-2-карбоксилата;
ж) метил-4-(2-циано-1(R₂)-этил)-3-[N-(диметиламинометилен)амино]-1Н-пирроло-2-карбоксилат обрабатывают метанольным раствором аммиака с получением 3(R₂)-3-[4-оксо-3H, 5H-пирроло[3,2-d] пиримидин-7-ил] пропаннитрила. Более подробно способ для получения соединений формулы

где R₁ аминогруппа (способ A), проводится следующим образом.The method of obtaining derivatives of 7 (CHR ₂ R ₃ ) -3H, 5H-pyrrolo- [3,2-d] pyrimidin-4-one of the formula (I ") (method B)

where R ₁ H;
R ₂ phenyl unsubstituted or substituted by halogen, or cyclohexyl;
R ₃ CH ₂ CN, consists in the fact that carry out sequential stages:
a) the reaction of the corresponding cyclic aldehyde with cyanoacetic acid in the presence of ammonium acetate to obtain 3 (R ₂ ) -substituted pentanedinitrile;
b) reaction of the obtained 3 (R ₂ ) -pentanedinitrile with formic acid alkyl ester in the presence of a base to obtain 3 (R ₂ ) -2-formylpentanedinitrile;
c) the latter is reacted with glycine methyl ester hydrochloride in the presence of sodium or ammonium acetate to obtain methyl N - [(3 (R ₂ ) -2,4-dicyano) -2-butenyl] glycine;
d) the obtained methyl-N - [(3 (R ₂ ) -2,4-dicyano) -2-butenyl] glycine is reacted with chloroacetic acid alkyl ester in the presence of 1,5-diazabicyclo [4.3.0] non-5- ene or 1,8-diazabicyclo [5.4.0] undec-7-ene to give methyl-3-amino-4- (2-cyano-1 (R ₂ ) -ethyl) -1-ethyl-1H-pyrrole-1 2-dicarboxylate;
d) methyl 3-amino-4- (2-cyano-1 (R ₂ ) -ethyl) -1-ethyl-1H-pyrrole-1,2-dicarboxylate is treated with a base to obtain methyl-3-amino-4- ( 2-cyano-1 (R ₂ ) ethyl) -1H-pyrrolo-2-carboxylate;
e) then methyl-3-amino-4- (2-cyano-1 (R ₂ ) -ethyl) -1H-pyrrolo-2-carboxylate is reacted with dimethylformamide dimethylacetal to give methyl 4- (2-cyano-1 ( R ₂ ) -ethyl) -3- [N- (dimethylamino-methylene) amino] -1H-pyrrolo-2-carboxylate;
g) methyl 4- (2-cyano-1 (R ₂ ) -ethyl) -3- [N- (dimethylaminomethylene) amino] -1H-pyrrolo-2-carboxylate is treated with a methanol solution of ammonia to obtain 3 (R ₂ ) - 3- [4-oxo-3H, 5H-pyrrolo [3,2-d] pyrimidin-7-yl] propannitrile. In more detail, a method for producing compounds of the formula

where R ₁ amino group (method A) is carried out as follows.

The first step of the method involves reacting the corresponding R ₂ -substituted cyclic aldehyde with cyanacetic acid at a molar ratio of about 1: 1 to 1: 5 in the presence of ammonium acetate at a temperature near the boiling point under reflux for about 10 hours to 8 days to obtain 3R ₂ -substituted pentanedinitrile as an intermediate. In a second step, the 3 (R ₂ ) -substituted pentanedinitrile is reacted with formic acid alkyl ester, for example formic acid ethyl ester, in the presence of a strong base, for example, a metal containing base, sodium hydride or sodium alkoxide, in particular sodium methoxide, at a molar ratio of about 1-2: 3-6: 1-3 and at a temperature of about 20-65 ^o C for about 10 hours to 8 days to obtain 3 (R ₂ ) -2-formylpentanedinitrile. The next step involves the interaction of 3 (R ₂ ) -2-formylpentanedinitrile with glycine methyl ester hydrochloride and in the presence of sodium or ammonium acetate at a molar ratio of about 1-2: 1.5-4: 1.5-4 and at a temperature of about 20- 60 ^° C. for about 10-48 hours to give methyl N - [(3 (R ₂ ) -2,4-dicyano) -2-butenyl] glycine as an intermediate. In the next step, methyl-N - [(3 (R ₂ ) -2,4-dicyano) -2-butenyl] glycine is reacted with chloroformic acid alkyl ester, for example, chloroformic acid ethyl ester, in the presence of 1,5-diazabicyclo [4.3 .0] non-5-eno (DBN) or 1,8-diazabicyclo [5.4.0] undec-7-eno (DBU) with a molar ratio of about 1-2: 1.5-5: 1.5-4 and at a temperature of about 0-50 ^o C for about 10 hours to 10 days to obtain methyl-3-amino-4- (2-cyano-1 (R ₂ ) ethyl) -1-ethyl-1H-pyrrole-1,2 -dicarboxylate as an intermediate. The next step involves the interaction of methyl 3-amino-4- (2-cyano-1 (R ₂ ) ethyl) -1-ethyl-1H-pyrrole-1,2-dicarboxylate with a base, for example, sodium carbonate, in a molar ratio of about 2: 1 to 1: 5 and at a temperature near room temperature for about 10-48 hours to obtain methyl 3-amino-4- (2-cyano-1 (R ₂ ) ethyl) -1H-pyrrole-2-carboxylate in as an intermediate compound. In the next step, methyl 3-amino-4- (2-cyano-1 (R ₂ ) -ethyl) -1H-pyrrole-2-carboxylate reacts with benzoylisothiocyanate in a molar ratio of about 2: 1 to 1: 2 and at a temperature of about room temperature for about 30 minutes to 3 hours to obtain N-benzoyl-N- [4- (2-cyano-1 (R ₂ ) ethyl) -2-methoxycarbonyl-1H-pyrrol-3-yl] thiourea as an intermediate connections. The next step in the interaction of N-benzoyl-N- [4- (2-cyano-1 (R ₂ ) ethyl) -2-methoxycarbonyl-1H-pyrrol-3-yl] thiourea with alkyl halide, for example methyl iodide, is carried out with molar a ratio of about 1: 1 to 1: 6 and at a temperature of about 0-30 ^o C for about 10 minutes to 10 hours to obtain N-benzoyl-N- [4- (2-cyano-1 (R ₂ ) -ethyl ) -2-methoxycarbonyl-1H-pyrrol-3-yl] -S-methylthiourea as an intermediate. In the next step, N-benzoyl-N- [4- (2-cyano-1 (R ₂ ) ethyl) -2-methoxycarbonyl-1H-pyrrol-3-yl] -S-methylthiourea (about 1-2 mol) is treated a methanol or ethanol solution of ammonia in a molar ratio of about 1: 1 to 1:20 and at a temperature of about 20-130 ^o C for about 16-60 hours to obtain a mixture of 2-amino compounds of the invention 3 (R ₂ ) -3- (2- amino-4-oxo-3H, 5H-pyrrolo [3,2-d] pyrimidin-7-yl) propannitrile and 3 (R ₂ ) -3- (2-methylmercapto-4-oxo-3H, 5H-pyrrolo [3 , 2-d] pyrimidin-7-yl) propannitrile, which can be used as an intermediate to produce another substance of the invention.

In another method for producing compounds, wherein R ^{1 is} hydrogen (method B) methyl-3-amino-4- (2-cyano-1 (R ₂ ) ethyl) -1H-pyrrole-2-carboxylate, obtained above in method A, treated with dimethylformamide dimethylacetal at a molar ratio of about 1: 1 to 1: 4 and at a temperature of about 25-100 ^o C for about 1-10 days to obtain methyl 4- (2-cyano-1 (R ₂ ) ethyl) -3 - [N- (dimethylaminomethylene) amino] -1H-pyrrolo-2-carboxylate as an intermediate. The next step involves the treatment of methyl 4- (2-cyano-1 (R ₂ ) ethyl) -3- [N- (dimethylaminomethylene) amino] -1H-pyrrolo-2-carboxylate with a methanol or ethanol ammonia solution in a molar ratio of about 1: 1 to 1:20 and at a temperature of about 20-130 ^o C for about 10-68 hours to obtain the compounds of the invention 3 (R ₂ ) -3- (4-oxo-3H, 5H-pyrrolo [3,2-d] pyrimidin-7-yl) propanenitrile.

 So, to obtain the compounds of formula (I) use 3-substituted propionitriles as starting materials. Such starting materials can be obtained using a variety of methods that are well known in the literature. The compound of formula (I) is then obtained from the specified starting material using a number of known reactions (MILim, RSKlein and JJ Fox, J. Org. Chem. 44.3826 (1979); MILim, RSKlein and JJFox, Tetrahedron Lett . 21,1013 (1980); MILim and RSKlein, Tetrahedron Lett. 22.25 (1981); MI Lim, WIRen, BAOtter and RSKlein, J. Org. Chem. 48,780 (1983)).

To obtain a group of compounds of formula (II), where R is pyridinyl, 3- (pyridinyl) propionitrile is used as the starting material. The corresponding 3- (pyridinyl) propionitrile can be obtained by converting the corresponding 3- (pyridinyl) propionyl chloride to the corresponding amide by amminolysis, for example, by treating ammonium, which is then dehydrated to the desired nitrile by distillation with a dehydrating agent, for example, POCl ₃ or SOCl ₂ . Either the starting material is obtained by condensation of 3-aldehyde with cyanoacetic acid followed by decarboxylation to obtain the corresponding substituted acrylonitrile, which is hydrogenated to obtain the corresponding 3- (pyridinyl) propionitrile either catalytically or by dissolving magnesium metal in methanol at 0 ^° C (Profitt, J. et al. J. Org. Chem. 40, 127 (1975)). Subsequently, substances (II) are obtained from the indicated substance according to the procedure (MILim, RSKlein and JJFox, J. Org. Chem. 44.3826 (1979); MI Lim, RSKlein and JJFox, Tetrahedron Lett. 21,1013 (1980); MILim and RSKlein, Tetrahedron Lett. 22.25 (1981); MILim, WIRen, BAOtter and RSKlein, J. Org. Chem. 48, 780 (1983)).

 It is possible to use cyclohexenylacetonitrile as the starting material for the preparation of compounds of group (III) as a result of the reaction of this starting material using the methodology (MILim, RSKlein and JJFox, J. Org. Chem. 44.3826 (1979); MILim, RS Klein and JJFox, Tetrahedron Lett. 21,1013 (1980); MILim and RSKlein, Tetrahedron Lett. 22,25 (1981); MILim, WIRen, BAOtter and RSKlein, J. Org. Chem. 48 780 (1983)). The catalytic hydrogenation of one of (IIIA), (IIIB) or (IIIC) gives the substance (IIID).

 A method of obtaining a group of compounds (IV) of the invention uses the corresponding 3-substituted propionitriles as starting materials. Such starting materials can be obtained by a number of methods that are well known in the literature. Substance (IV) is then prepared from this starting material using techniques (MILim, RSKlein and JJFox, J. Org. Chem. 44.3826 (1979); MILim, RSKlein and JJFox, Tetrahedron Lett. 21, 1013 (1980); MILim and RSKlein, Tetrahedron Lett. 22.25 (1981); MI Lim, WIRen, BAOtter and RSKlein, J. Org. Chem. 48, 780 (1983)).

 As is obvious to specialists in this field of technology, variants of the above methods are useful in obtaining a number of substances of the invention without deviating from the invention. Known methods are used for reactions involving certain intermediates requiring the protection of nitrogen or oxygen atoms in the intermediates.

 To more fully describe the invention, the following examples are given without limiting the scope of its application. In the examples, all parts and percentages are given in mass. unless otherwise specified. The proportions of solvent mixtures used as chromatographic solvents for elution are given by volume.

 Tests (as follows from the application materials) were conducted on mammals. In particular, in the examples given in the description, the experiments were carried out on rats of the Lewis strain, which received intraperitoneal administration of the substances obtained according to the invention in the form of injection solutions, indicating dosages, frequency of injections and comparative experiments with control groups of animals. The results are specified there (examples 22, 53, 65, 70, 81, 89, etc.), as well as in the table. one.

Example 1. A method of obtaining 3- (3-pyridinyl) propionitrile. Argon is introduced into the three-necked flask equipped with a magnetic stirrer, a thermometer, a dropping funnel and a reflux condenser. Freshly ground potassium hydroxide (6.6 g, 0.1 mol) and anhydrous acetonitrile (150 ml) are charged into a flask and heated to boiling under reflux, 3-pyridinecarboxaldehyde (10.7 g, 0.1 mol) in anhydrous acetonitrile is added (50 ml) dropwise over about 5 minutes and refluxing continued for the next about 3 minutes. The resulting hot reaction mixture was poured into ice / water (100 g) and the resulting solution was extracted with CH ₂ Cl ₂ (3 x 100 ml), dried with Na ₂ SO ₄ and evaporated to give crude 3- (3-pyridinyl) acrylonitrile, which is purified by column chromatography over silica gel using CHCl ₃ as an elution solvent; yield 3.3 g (25.6%).

In an argon atmosphere, a stirred solution of acrylonitrile (2.662 g, 0.02 mol) in 99% ethanol (100 ml) was treated with a drop of 4% aqueous sodium hydroxide, then sodium borohydride (0.378 g, 0.01 mol). Additionally, sodium borohydride (0.378 g) was added twice more at 4-hour intervals. The mixture was stirred at room temperature overnight, diluted with water, extracted with EtOAc and dried with Na ₂ SO ₄ . The solvent was removed under reduced pressure, the resulting crude product was chromatographed over a silica gel column using chloroform: methanol (40: 1) as an elution solvent to obtain 2.2 g (84.6%) of the product as a colorless oil.

 The 3- (3-pyridinyl) propionitrile obtained in Example 1 is then used to synthesize the compounds of the invention.

Example 2. In an atmosphere of dry N _{2, a} mixture of 3- (3-pyridinyl) propionitrile (0.661 g, 5.0 mmol), sodium hydride (0.240 g, 10.0 mmol) and formic acid ethyl ester (1.11 g, 15 , 0 mmol) in anhydrous tetrahydrofuran (20 ml) was stirred for 48 hours with protection from air and moisture. The volatile material was evaporated and a solution of the solid residue in 15 ml of cold water was adjusted at 0 ^° C. to pH 6 with cold 6N HCl. The resulting oily mixture was extracted with CHCl ₃ , the extract was washed with water, dried using Na ₂ SO ₄ and evaporated to give a dark oil, which is a mixture of 2-formyl-3- (3-pyridinyl) propionitrile and nitrile starting material. This crude product is used in the next reaction without further purification.

Example 3. Glycine methyl ester hydrochloride (0.942 g, 7.5 mmol) and anhydrous sodium acetate (0.615 g, 7.5 mmol) are added to a solution of crude formyl derivative (0.89 g) in MeOH / H ₂ O (4: 1.50 ml). After 24 hours, MeOH was evaporated in vacuo, the mixture of water and oil was extracted with CHCl ₃ . The CHCl ₃ layer was dried (Na ₂ SO ₄ ) and evaporated to give an amber oil, which was sent to a silica gel column. Elution with CHCl ₃ gives two main layers: 3- (3-pyridinyl) propionitrile (used as starting material in the previous step); desired enamine.

Example 4. In a nitrogen atmosphere, ethyl chloroacetic acid (0.521 g, 4.8 mmol) was added dropwise to a solution of enamine (example 3) (0.513 g, 3.2 mmol) and 1,5-diazabicyclo [4.3.0] non -5-ena (DBN, 1.37 g, 11.1 mmol) in dry CH ₂ Cl ₂ (15 ml) with external cooling in an ice bath. After stirring at 0 ^° C. for 1 h, the solution was left at room temperature overnight. After checking the progress of the reaction using TLC, an additional amount of ClCO ₂ Et (0.1 ml) and DBN (1.0 g) were added to complete the reaction, the solution was kept for 24 hours. The volatile material was evaporated in vacuo and the viscous residue was purified in a short column silica gel (the main purpose of which is to remove a less mobile DBN), obtaining N-protected pyrrole, which is used for the next step without further purification.

Example 5. To a solution of the N-protected pyrrole of Example 4 (0.635 g, 2.0 mmol) in MeOH (50 ml) was added solid Na ₂ CO ₃ (0.212 g, 2.0 mmol) and the reaction mixture was stirred at room temperature for 48 h with the release of the resulting unlocked pyrrole. The mixture was evaporated to dryness and the residue was triturated with H ₂ O (25 ml) to dissolve the inorganic substances and extracted with CHCl ₃ (3 x 100 ml). The extract was dried (Na ₂ SO ₄ ) and evaporated to give a viscous resin, which crystallized upon drying in vacuo to be used as an intermediate without further purification. Higher purification can be carried out either by column chromatography using silica gel / CHCl ₃ , or by recrystallization from toluene / cyclohexane (1: 3).

Example 6. Benzoylisothiocyanate (0.232 g, 1.42 mmol) was added dropwise to a solution of the unblocked pyrrole of Example 5 (0.290 g, 1.18 mmol) in dry CH ₂ Cl ₂ (100 ml). After 1 h at room temperature, the solution was evaporated and the gummy residue was stirred in Et ₂ O / cyclohexane (1: 1.20 ml). The resulting yellow solid suspension was filtered off under nitrogen pressure, and the thioureide compound was dried in vacuo over P ₂ O ₅ .

Example 7. Methyl iodide (0.228 g, 1.61 mmol) was added to a solution of the thiouride of Example 6 (0.383 g, 0.94 mmol) and DBN (0.140 g, 1.12 mmol) in dry CH ₂ Cl ₂ (10 ml) at 0 ^° C. The solution was stirred at 0 ^° C. for 15 minutes at ambient temperature for 1 hour and then evaporated in vacuo. A solution of the residue in CHCl _{3 was} chromatographed on a silica gel column with a mixture of CHCl ₃ / methanol as an elution solvent to obtain homogeneous fractions of a methylthio substituted intermediate.

 Example 8

Раствор метилтиосоединения примера 7 (0,358 г, 0,85 ммоль) в 100 мл MeOH, который был насыщен NH₃ при 0^oC, нагревают при 90-95^oC в течение 24 ч в футерованной стеклянными плитками бомбе из нержавеющей стали. Содержимое охлажденной бомбы выпаривают в вакууме, получая смесь желаемого 2-аминопроизводного, бензамида и побочного продукта, представляющего собой 2-метилтиопроизводное. Смесь растворяют в метаноле и раствор выпаривают с силикагелем (около 5 г). Смесь затем осторожно наносят на верхнюю часть хроматографической колонки с силикагелем, которую затем элюируют смесью CHCl₃/MeOH (9: 1), получая побочное метилтиосоединение и желаемый 2-амино-7-(3-пиридинилметил)-3H, 5H-пирроло[3,2-d]пиримидин-4-он, как промежуточное соединение. Дальнейшую очистку проводят перекристаллизацией из кипящего изопропилацетата в аппарате Сокслета.

A solution of the methylthio compound of Example 7 (0.358 g, 0.85 mmol) in 100 ml of MeOH that was saturated with NH ₃ at 0 ^° C was heated at 90-95 ^° C for 24 hours in a glass-lined stainless steel bomb. The contents of the chilled bomb were evaporated in vacuo to give a mixture of the desired 2-amino derivative, benzamide and a 2-methylthio derivative by-product. The mixture was dissolved in methanol and the solution was evaporated with silica gel (about 5 g). The mixture was then carefully applied to the top of a silica gel chromatography column, which was then eluted with a CHCl ₃ / MeOH (9: 1) mixture to obtain the methyl thio side compound and the desired 2-amino-7- (3-pyridinylmethyl) -3H, 5H-pyrrolo [3 , 2-d] pyrimidin-4-one, as an intermediate. Further purification is carried out by recrystallization from boiling isopropyl acetate in a Soxhlet apparatus.

Analysis: calculated for C ₁₂ H ₁₁ N ₅ O: C 59.74; H 4.60; N, 29.03; Found: C, 59.58; H 4.86; N, 28.89.

MS (FAB): 242 (M + H) ⁺ ; UV: 0.1 N HCl 233 (18.2), 269 (17.4); pH 7,233 (18.2); 269 (17.4); pH 7,233 (22.4); 269 (12.3); 0.1 N NaOH 227 (22.2), 263 (10.5), 290 (sh). IC ₅₀ (PNP) 25 nM.

Раствор пиридинилметильного промежуточного соединения в 0,1N HCl гидрируют на платиновом катализаторе при давлении H₂ 60 фунтов/дюйм² (4,2186 кг/см²). Катализатор генерируют посредством кратковременного гидрирования PtO₂ в 0,1N HCl. Когда реакция заканчивается, катализатор удаляют фильтрованием под давлением N₂ и фильтрат выпаривают. Остаток в минимальном количестве этанола разбавляют медленно большим количеством EtO₂ и получают гидрохлоридную соль в виде белого гигроскопичного твердого вещества (IC). Т.пл. (в ^oС) при 200^o размягчается.

A solution of intermediate piridinilmetilnogo in 0,1N HCl was hydrogenated over platinum catalyst under pressure of H ₂ 60 pounds / inch ² (4.2186 kg / cm ^2). The catalyst is generated by briefly hydrogenating PtO ₂ in 0.1 N HCl. When the reaction is complete, the catalyst is removed by filtration under N ₂ pressure and the filtrate is evaporated. The residue in a minimum amount of ethanol is diluted slowly with a large amount of EtO _{2 to} give the hydrochloride salt as a white hygroscopic solid (IC). Mp (in ^o C) at 200 ^o softens.

MS (FAB) m / e 248 (M + H) ⁺ ; UV 0.1 N HCl 234 (15.7), 273 (13.8) pH 7 buffer 232 (19.1), 271 (9.3), 0.1 N NaOH 229 (19.0), 266 ( 7.1), 288 (sh); ¹ H-NMR (DNSO-d ₆ ) δ 1.19, 1.61, 1.77 (m, 4H, H-4's and 5's piperidine), 1.96 (m, 1H, H-3 piperidine), 2 53 (d, 2H, -CH ₂ -), 2.69 (m, 2H, H-2's piperidine), 3.14 (d, 2H, H-6's piperidine), 7.25 (d, J 2, 8, 1H, H-6), 7.76 (s, 2H, NH ₂ ), 8.81, 9.18 (br 1 each, NH ^{+ is} exchanged with D ₂ O), 12.24 (s, 1H S-NH).

Analysis: calculated for C ₁₂ H ₁₇ N ₆ O • 2.8HCl • 0.2 C ₂ H ₆ O • H ₂ O: C, 39.55; H, 6.16; N, 18.60; Cl, 26.36; Found: C, 39.75; H 6.24; N, 18.65; Cl 26.08.

IC ₅₀ 1 μM.

Example 9. The substance (IC) of example 8 is tested for enzymatic inhibitory activity. An enzymatic analysis of purine nucleoside phosphorylase (PNP) is carried out, in which PNP activity (IC ₅₀ ) is studied for a substance that is determined radiochemically by measuring the formation of [ ¹⁴ C] -poxanthine from [ ¹⁴ C] -inosine (Biomedicina, 33.39 (1980)), using the spleen of the back of the leg as a source of enzyme.

Examples 10-14. The following substances of the invention, which are 2-amino-7- (CH ₂ R) -3H, 5H-pyrrolo [3,2-d] pyrimidin-4-ones, in which the R group means the following:
Example 10: R 2-methyl-3-piperidinyl;
Example 11: R 2-chloro-3-piperidinyl;
Example 12: R 2-trifluoromethyl-3-piperidinyl;
Example 13: R 2-methoxy-3-piperidinyl;
Example 14: R 2-fluoro-3-piperidinyl
receive, following the methods described in examples 1-8, using the corresponding 3- (substituted 3-pyridinyl) propionitriles as starting materials.

Example 15. 3- (2-Pyridinyl) propionitrile was prepared in this example using the method (V. Boekelheide, et al. J. Am. Chem. Soc. 75, 3243 (1953)). A solution of potassium cyanide (83.74 g) in water (160 ml) was added to a solution of freshly distilled 2-vinylpyridine (67.59 g) in acetic anhydride (131.30 g) at a speed adjusted to maintain a gentle boil under reflux. When the addition is complete, the resulting dark red mixture is heated for about 17 hours at 105 ^° C. in an oil bath with vigorous stirring. The cooled reaction mixture was adjusted to pH 8 with a saturated solution of Na ₂ CO ₃ . The mixture was extracted with CHCl ₃ (4 • 150 ml), washed with water, dried with Na ₂ SO ₄ and evaporated to a dark viscous oil. The oil is fractionally distilled in vacuo through a Vigreaux column. After a small pregon containing 2-vinylpyridine (0.35 g), the desired 3- (2-pyridinyl) propionitrile was collected as a clear, fluorescent yellow-green viscous oil: yield 59.8 g; 70.4% t. 86 ^o C at 1.0 mmHg.

Examples 16-20. The following compounds of the invention are obtained: 2-amino-7- (CH ₂ R) -3H, 5H-pyrrolo [3,2-d] pyrimidin-4-ones, in which the R group has the following meanings:
Example 16: R 2-piperidinyl;
Example 17: R 4-piperidinyl;
Example 18: R 3-trifluoromethyl-4-piperidinyl;
Example 19: R 3-methoxy-2-piperidinyl;
Example 20: R 3-fluoro-4-piperidinyl
following the methods described in examples 1-8 and 15, using the corresponding 3- (2- or 4-pyridinyl) propionitriles as starting materials.

 Example 21. A pharmaceutical composition for intraperitoneal injection is prepared for testing a substance (IC). For this, an intraperitoneal injection solution containing the substance of Example 8 is dissolved in an aqueous vehicle that contains 10% DMSO.

Example 22. Intraperitoneally injected into Lewis Rats the composition of example 21 containing a substance (IC) with a dose of 30 mg of a substance (IC) with an injection twice a day. Use control groups that receive only filler. After a certain time after administration, animals are euthanized and plasma samples are obtained. The plasma is extracted with cold 0.5N HClO ₄ and neutralized with solid NH ₄ HCO ₃ . After removal of the perchlorate salts, the extract was subjected to HPLC (high pressure liquid chromatography) on a reversed phase column (Spherisorb ODSI). A significant increase in plasma inosine is observed in plasma taken from animals receiving the substance (IC).

 Examples 23-33. The substances obtained in examples 10-20 were used to prepare drugs in accordance with the method of preparation of example 21. The resulting spray solutions were tested in accordance with the method of example 22. A significant increase in plasma inosine was observed in plasma taken from animals receiving the substances of the invention.

Example 34. Receive 3- (2-furanyl) propionitrile. In a three-necked flask equipped with a condenser and a drain tube, magnesium chips (20 g) are added to a solution of 3- (2-furanyl) acrylonitrile (67.2 g) in dry methanol (2 L). An additional amount of magnesium is added in portions depending on the reaction rate until the entire amount (145 g) has been added. After about 5 hours, the solvent was evaporated until the contents formed a solid paste, which was adjusted to a pH of about 6.5 with 6N HCl under cooling. The mixture was extracted with several portions of CHCl ₃ , dried with Na ₂ SO ₄ and concentrated to a dark oil. Distillation in vacuo through a short Vigreaux column gives propionitrile as a colorless oil; yield 49.5 g (72%); t. 46.0-46.5 ^o C at 0.5 mmHg.

 The 3- (2-Furanyl) propionitrile from Example 34 is then used in syntheses of the invention.

Example 35. In an atmosphere of dry N _2, propionitrile (48.7 g), sodium hydride (10.28 g), formic acid ethyl ester (32.74 g) and anhydrous tetrahydrofuran (200 ml) were stirred at room temperature with moisture protection for 18 hours. The volatile substance is then evaporated, the resulting yellow solid is dissolved in about 20 ml of cold water with cooling in an ice bath, the solution is adjusted to pH 6.0 with cold 6N HCl. The resulting heavy oily precipitate was extracted into CHCl ₃ and the extract was washed with water, dried with Na ₂ SO ₄ and evaporated to give a liquid oil that contains a crude formyl derivative; yield 53.06 g.

Example 36. Glycine methyl ester hydrochloride (67.05 g) and anhydrous sodium acetate (43.79 g) were added to a solution of the crude formyl derivative of the previous example (53.05 g) in MeOH / H ₂ O (4: 1.1500 ml ) After 24 hours, MeOH was evaporated in vacuo and the mixture of water and oil was extracted with CHCl ₃ . The CHCl ₃ layer was dried (Na ₂ SO ₄ ) and evaporated to give an amber oil (59.1 g).

Example 37. In a nitrogen atmosphere, chloroformate ethyl ester (43.68 g) was added dropwise to a solution of the amber oil of Example 36 (59.1 g) and DBN (133 g) in dry CH ₂ Cl ₂ (400 ml) with external cooling in an ice bath. After stirring at 0 ^° C. for 1 h, the solution was allowed to stand at room temperature overnight. After checking the course of the reaction using TLC, an additional amount of ClCO ₂ Et and DBN are introduced to complete the reaction and the solution is left to stand for 24 hours. The volatile material is evaporated in vacuo, the viscous residue is purified on a silica gel column (the main purpose of which is to remove less mobile DBN), obtaining N-protected pyrrole, the corresponding pyrrole without an ethoxycarbonyl protecting group on pyrrole nitrogen and the starting propionitrile.

Example 38. To a solution of the N-protected pyrrole of Example 37 (11.66 g) in MeOH (200 ml), solid Na ₂ CO ₃ (4.23 g) was added and the reaction mixture was stirred at room temperature for 24 hours to separate the resulting unlocked pyrrole. The mixture was evaporated to dryness and the residue was thoroughly triturated with H ₂ O (200 ml) to dissolve the inorganic substances, then extracted with CHCl ₃ (3 x 200 ml). The extract was dried (Na ₂ SO ₄ ) and evaporated to give a viscous resin which crystallized upon drying in vacuo to be used as an intermediate without further purification. Deeper purification can be carried out by column chromatography using silica gel / CHCl ₃ , or by recrystallization from toluene / cyclohexane (1: 3).

Example 39. Benzoylisothiocyanate (4.58 g) was added dropwise to a solution of the unblocked pyrrole of Example 38 (5.15 g) in dry CH ₂ Cl ₂ (75 ml). After 1 h at room temperature, the solution was evaporated, the gummy residue was dissolved in Et ₂ O (100 ml) with an almost immediate release of crystalline solid, which was collected by filtration. The filtrate Et ₂ O is heated to boiling and diluted with an equal volume of hot cyclohexane. With slow cooling, the solution gives an additional amount of a thioreid derivative.

Example 40. Methyl iodide (8.70 g) was added to a solution of the thioureide derivative of Example 39 (10.68 g) and DBN (4.15 g) in dry CH ₂ Cl ₂ (250 ml) at 0 ^° C. The solution was stirred at 0 ^o C for 15 min, at ambient temperature for 1 h and then evaporated in vacuo. A solution of the residue in CHCl _{3 was} chromatographed on a silica gel column using CHCl ₃ as an elution solvent to obtain homogeneous fractions of a methylthiomethane intermediate.

 Example 41

Раствор метилтиосоединения из примера 40 (0,80 г, 2,0 ммоль) в 25 мл MeOH, который был насыщен NH₃ при 0^oC, нагревают при 90-95^oC в течение 24 ч в футерованной стеклянными плитками бомбе из нержавеющей стали. Содержимое охлажденной бомбы выпаривают в вакууме, получая смесь 2-аминосоединения, бензамида и побочного продукта, который является 2-метилтиопроизводным. Смесь перемешивают энергично в течение нескольких минут с 30 мл смеси 2:1 Et₂O/циклогексан, и нерастворимое белое твердое вещество отфильтровывают и промывают Et₂O. Фильтрат содержал большую часть бензамида и 2-метилтиопроизводного. Раствор нерастворимого в смеси Et₂O/циклогексан твердого вещества (0,425 г) в MeOH выпаривают с приблизительно 10 г силикагеля. Измельченный в порошок остаток наносится равномерно на верх колонны с силикагелем, которая затем элюируется с помощью смеси CHCl₃/MeOH/HOAc (95:5:1), давая 2-метилтиоипроизводное как побочный продукт и желаемое фуранильное промежуточное соединение. Промежуточное соединение перекристаллизовывают экстракцией в кипящем изопропилацетате в аппарате Сокслета. Белые кристаллы собирают в три порции и сушат в вакууме над P₂O₅ при 110^oC в течение 7 ч.

A solution of the methylthio compound of Example 40 (0.80 g, 2.0 mmol) in 25 ml of MeOH that was saturated with NH ₃ at 0 ^° C was heated at 90-95 ^° C for 24 hours in a glass-lined stainless steel bomb . The contents of the cooled bomb were evaporated in vacuo to give a mixture of the 2-amino compound, benzamide and a by-product that is 2-methylthio derivative. The mixture was stirred vigorously for several minutes with 30 ml of a 2: 1 Et ₂ O / cyclohexane mixture, and the insoluble white solid was filtered off and washed with Et ₂ O. The filtrate contained most of the benzamide and 2-methylthio derivative. A solution of a solid insoluble in a mixture of Et ₂ O / cyclohexane (0.425 g) in MeOH was evaporated with approximately 10 g of silica gel. The powder residue is uniformly applied to the top of the silica gel column, which is then eluted with a mixture of CHCl ₃ / MeOH / HOAc (95: 5: 1) to give the 2-methylthio derivative as a by-product and the desired furanyl intermediate. The intermediate is recrystallized by extraction in boiling isopropyl acetate in a Soxhlet apparatus. White crystals are collected in three portions and dried in vacuo over P ₂ O ₅ at 110 ^o C for 7 hours

A solution of the intermediate (116 g, 0.5 mmol) in methanol (50 ml) was hydrogenated with 30% palladium-charcoal (40 mg) at a pressure of H ₂ 62 psi (4.3592 kg / cm) in for about 36 hours. The catalyst was removed by filtration under a pressure of N ₂ and the filtrate was evaporated, then re-evaporated with toluene. The solid residue is recrystallized by extraction in boiling isopropyl acetate in a Soxhlet apparatus. 2-Tetrahydrofuranyl compound (IE) is obtained as a white crystalline solid, which is dried in vacuo over P ₂ O ₅ at 110 ^° C. for about 6 hours; yield 73 mg (61.8%); so pl. 284-286 ^o C (decomp.).

MS (FAB) m / e 235 (M + H) ⁺ , UV 0.1 N HCl 235 (17.1), 273 (14.7), pH 7 buffer 232 (20.0), 271 (10.5 ), 0.1 N NaOH 228 (20.2), 264 (7.0), 286 (6.2); ¹ H-NMR (DMSO-d ₆ ) δ 1.50, 1.78 (m, 4H, H-3's and 4's tetrahydrofuranyl), 2.52, 2.69 (2d, 2H, -CH ₂ R), 3 57, 3.76 (2m, 1H each, H-5's tetrahydrofuranyl), 5.82 (s, 2H, NH ₂ ), 6.97 (d, J = 2.8, 1H, H-6), 10 34 (s, 1H, 3-NH); 11.18 (s, 1H, 5-NH).

Analysis: calculated for C ₁₁ H ₁₄ N ₄ O ₂ : C 56.40; H 6.02; N, 23.92; Found: C, 56.33; H 6.49; N, 23.95.

Example 42. The product of example 41 was tested for enzymatic inhibitory activity in accordance with the method of example 9, and PNP activity (IC ₅₀ 11 μM) was observed for the substance.

 Example 43. Prepare a pharmaceutical composition for intraperitoneal injection for testing substances (IE). For this, an intraperitoneal injectable solution containing the substance of Example 41 is dissolved in an aqueous vehicle that contains 10% DMSO.

 Example 44. Using the method of Example 16, the composition of Example 43 containing the substance (IE) with a dose of 30 mg of the substance (IE) was administered intraperitoneally in Lewis Rats, and the results were analyzed in comparison with the control. A significant increase in plasma inosine is observed in plasma taken from animals receiving the substance (IE).

Examples 45-49. The following compounds of the invention are obtained: 2-amino-7- (CH ₂ ) -3H, 5H-pyrrolo [3,2-d] pyrimidin-4-ones, in which the group R means:
Example 45: R 3-tetrahydrofuranyl;
Example 46: R 3-chloro-2-tetrahydrofuranyl;
Example 47: R 3-trifluoromethyl-2-tetrahydrofuranyl;
Example 48: R 3-methoxy-3-tetrahydrofuranyl;
Example 49: R 3-fluoro-2-tetrahydrofuranyl.

 Substances obtained by following the methods described in examples 34-41, using the corresponding 3- (furanyl) -acrylonitriles as starting materials.

 Example 50

В атмосфере азота раствор тетрагидрофуранильного производного (IE) (500 мг), полученного в примере 41, и кристалл фенола в 2N HBr (20 мл) перемешивают в течение 18 ч при 40^oC. Растворитель выпаривают в вакууме при низкой температуре, остаток промывают с помощью нескольких мл Et₂O, декантацией удаляя любой трибромфенол. Т.пл. 311-313^oC.

Under nitrogen atmosphere, a solution of the tetrahydrofuranyl derivative (IE) (500 mg) obtained in Example 41 and a phenol crystal in 2N HBr (20 ml) were stirred for 18 h at 40 ^° C. The solvent was evaporated in vacuo at low temperature, the residue was washed with using a few ml Et ₂ O, decanting removing any tribromphenol. Mp 311-313 ^o C.

MS (FAB) m / e 251 (M + H) ⁺ ; UV 0.1 N HCl 234 (17.4), 274 (15.2), pH 7 buffer 232 (20.9), 272 (10.6), 0.1 N NaOH 228 (21.3), 265 (7.3), 287 (6.5); IR (KBr) 3211, 3203, 3127, 3108, 2936, 1661, 1609, 1567, 1524, 1369 cm ^-1 ; ¹ H-NMR (DMSO-d ₆ ) δ 1.62, 1.87, 2.01 (complex m, 4H, H-3'S and 4'S tetrahydrothienyl), 2.72 (m, 4H, -CH ₂ -overlapping H -5'S tetrahydrothienyl), 3.64 (p, 1H, 6-2 tetrahydrothienyl), 5.82 (s, 2H, NH ₂ ), 6.97 (d, J = 3.0, 1H, H-6), 10.34 (s, 1, 3-NH); 11.17 (s, 1.5-NH);
Analysis: calculated for C ₁₁ H ₁₄ N ₄ OS: C 52.78; H 5.64; N, 22.38; Found: C, 52.56; H 5.64; N, 22.50.

From the filtrate, 2-amino-1,5-dihydro-7- (1,2-dithian-3-ylmethyl) -4H-pyrrolo [3,2-d] pyrimidin-4-one; so pl. 311-313 ^o C.

MS (FAB) m / e 283 (M + H) ⁺ ; IR (KBr) 3139, 3135, 3131, 2926, 1673, 1658, 1609, 1569, 1523, 1370 cm ^-1 ; ¹ H-NMR (DMSO-d ₆ ) d 1.53, 1.67, 2.04 (complex m, 4H, H-4's and 5's of dithienyl), 2.67 (m, 2H, -CH ₂ R); 2.75 (m, 2H, H-6's of dithienyl), 3.17 (m, 1H, H'3 of dithienyl), 5.84 (s, 2H, NH ₂ , 7.00 (d, J = 3.0 , 1H, H-6), 10.34 (s, 1H, 3-NH), ll, 24 (s, 1H, 5-NH).

Analysis: calculated for C ₁₁ H ₁₄ N ₄ OS ₂ : C 46.80; H 5.00; N, 19.85; Found: C, 46.78; H 4.97; N, 19.81.

 The suspension of the residue in water was adjusted to pH 7 1N NaOH to give bromoalcohol as the free base. The solid is collected, washed with cold water and dried at room temperature.

A solution of bromoalcohol in anhydrous dimethylacetamide is cooled in an ice bath and treated with an equimolar portion of PBr ₃ . The solution was stirred at room temperature for 1 h, then dimethylacetamide was evaporated in vacuo at low temperature using an air pump and a dry ice trap. A suspension of the residue in ice / water was adjusted to pH 7 using 1N NaOH to give a crude reaction product containing 1,4-dibromo compound. After filtration, washing with cold water and drying at room temperature, the resulting product is refluxed with sodium sulfide in a mixture of 50% ethanol / water, or heated with Na ₂ S in a solution of N, N-dimethylacetamide; the solvent was removed in vacuo, the residue was washed with water to remove NaBr, and the pH was adjusted to about 7 to precipitate 2-tetrahydrothienyl substance (IG).

Example 51. The substance of Example 50 was tested for enzymatic inhibitory activity as in Example 9, and PNP activity (IC ₅₀ 11 nM) was observed for the substance. IC ₅₀ 18 nM for the dithian-3-yl compound.

 Example 52. Prepare a pharmaceutical composition for intraperitoneal injection for testing substances (IG). An intraperitoneal injection solution containing the substance of Example 50 is dissolved in an aqueous vehicle that contains 10% DMSO.

 Example 53. Using the method of Example 16, the experimental composition of Example 52 containing a substance (IG) with a dose of 30 mg of a substance (IG) was administered intraperitoneally in Lewis Rats, and the results were analyzed in comparison with the control. A significant increase in plasma inosine is observed in plasma taken from animals receiving the substance (IG).

Examples 54-58. The following compounds of the invention are obtained, 2-amino-7- (CH ₂ R) -3H, 5H-pyrrolo [3,2-d] pyrimidin-4-ones, in which R means:
Example 54: R 3-tetrahydrothienyl;
Example 55: R 3-chloro-2-tetrahydrothienyl;
Example 56: R 3-trifluoromethyl-2-tetrahydrothienyl;
Example 57: R 3-methoxy-2-tetrahydrothienyl;
Example 58: R 3-fluoro-2-tetrahydrothienyl.

 Substances obtained by following the methods described in examples 34-41 and 50, using the corresponding 3- (furanyl) -acrylonitriles as starting materials.

 Example 59. Using the method of Example 1, 3- (2-pyrrolyl) propionitrile was prepared using pyrrole-2-carboxaldehyde as the starting material. Following examples 2-8, the intermediate 2-amino-7- (2-pyrrolylmethyl) -3H, 5H-pyrrolo [3,2-d] pyrimidin-4-one is obtained from propionitrile, from which 2-pyrrolidinyl substance (II) obtained by reducing the intermediate.

 Example 60. Using the method of Example 1, 3- (3-pyrrolyl) propionitrile was prepared using pyrrole-3-carboxaldehyde as the starting material. Following examples 2-8, the intermediate 2-amino-7- (3-pyrrolylmethyl) -3H, 5H-pyrrolo [3,2-d] pyrimidin-4-one is obtained from propionitrile, from which 3-pyrrolidinyl substance (IJ) obtained by reducing the intermediate.

 Example 61. Using the method of Example 1, 3- (2-pyranyl) propionitrile was prepared using 2-pyrancarboxaldehyde as the starting material. Following examples 2-8, the intermediate 2-amino-7- (2-pyranylmethyl) -3H, 5H-pyrrolo [3,2-d] pyrimidin-4-one is obtained from propionitrile, from which 2-tetrahydropyranyl substance (IL) obtained by reducing the intermediate.

 Example 62. Using the method of Example 1, 3- (3-pyranyl) -propionitrile was prepared using 3-pyrancarboxaldehyde as the starting material. Following examples 2-8, the intermediate 2-amino-7- (3-pyranylmethyl) -3H, 5H-pyrrolo [3,2-d] pyrimidin-4-one is obtained from propionitrile, from which 3-tetrahydropyranyl substance (IM) obtained by reducing the intermediate compound.

 Example 63. Using the method of Example 1, 3- (4-pyranyl) propionitrile was prepared using 4-pyrancarboxaldehyde as the starting material. Following examples 2-8, the intermediate 2-amino-7- (4-pyranylmethyl) -3H, 5H-pyrrolo [3,2-d] pyrimidin-4-one is obtained from propionitrile, from which 4-tetrahydropyranyl substance (IN) obtained by reducing the intermediate compound.

Example 64. A solution of the methylthio derivative of Example 7 (0.358 g, 0.85 mmol) in 100 ml of MeOH that was saturated with NH ₃ at 0 ^° was heated at 90-95 ^° C for 24 hours in a glass-lined stainless steel bomb. The contents of the cooled bomb are evaporated in vacuo to give a mixture of the desired 2-amino intermediate, benzamide, and a by-product that is 2-methylthio derivative. The mixture was dissolved in methanol and the solution was evaporated with silica gel (about 5 g). The mixture was then carefully applied to the top of a silica gel chromatography column, which was then eluted with a CHCl ₃ / MeOH (9: 1) mixture to give a methylthioproducible product and the desired 2-amino-7- (3-pyridinylmethyl) -3H, 5H-pyrrolo [ 3,2-d] pyrimidin-4-one, substance (IIA). Further purification is carried out by recrystallization from boiling isopropyl acetate in a Soxhlet apparatus.

Example 65. The substance (IIA) of example 64 was tested for enzymatic inhibitory activity. An enzymatic assay of purine nucleoside phosphorylase (PNP) is carried out, in which PNP activity (IC ₅₀ ) is observed for a substance that is determined radiochemically by measuring the formation of [ ¹⁴ C] -poxanthine from [ ¹⁴ C] -inosine (Biomedicina, 33, 39, 1980) using a spleen the back of the leg as a source of enzyme.

Examples 66-67. The following compounds of the invention are obtained, 2-amino-7- (CH ₂ R) -3H, 5H-pyrrolo [3,2-d] pyrimidin-4-ones, in which R means:
Example 66: R 2-pyridinyl;
Example 67: R 3-chloro-2-pyridinyl.

 Substances are prepared following the methods given in Examples 1-7 and 64 using the corresponding 3- (pyridinyl) propionitriles as starting materials.

Метил 3-[(N-бензоил-S-метилизотиокарбамоил)амино]-4-[(4- пиридил)метил] -1H-пиррол-2-карбоксилат (BCX-180, 9,5 г, 23,25 ммоль) добавляют в 300-мл выложенный стеклянной футеровкой стеклянный сосуд высокого давления к 90 мл метилата аммония (полученного путем пропускания аммиака в безводный метанол при температуре 0-5^oC до насыщения) и реакционный сосуд закупоривают. Этот сосуд нагревают до температуры 85-90^oC в течение 12 ч и затем охлаждают до комнатной температуры. Летучие вещества выпаривают с помощью вращающегося испарителя и к полученному остатку добавляют 150 мл 1N HCl и 100 мл хлороформа. Полученную смесь энергично перемешивают и выделяют экстракт HCl. Экстракт HCl нейтрализуют, используя водный раствор карбоната натрия. Выпавшее в осадок твердое вещество выделяют путем фильтрации, высушивают на воздухе и экстрагируют 2 л этанола в аппарате Сокслета в течение 48 ч. Этанол выпаривают и образовавшийся светло-коричневый твердый остаток (3,1 г) размешивают с 20 мл деионизированной (DI) воды. Концентрированную (37%) соляную кислоту добавляют по каплям до тех пор, пока все твердое вещество не растворяется (2,1 мл, 25,7 ммоль). Прозрачный раствор обрабатывают активированным углем (700 мг) при температуре 50^oC в течение 5 мин и фильтруют через короткую колонку, заполненную цеолитом. Полученный фильтрат концентрируют на вращающемся испарителе при температуре 50- 55^oC с получением белого твердого вещества, высушивают сначало на воздухе, а затем в вакууме при температуре перегонки толуола. Масса полученного белого твердого вещества составляет 3,6 г (48%), т.пл. 340-342^oC.Example 68. 2-amino-4-oxo-3H, 5H-7 - [(4-pyridyl) methyl] pyrrolo [3,2-d] pyrimidine hydrochloride hydrate (2: 4: 1)

Methyl 3 - [(N-benzoyl-S-methylisothiocarbamoyl) amino] -4 - [(4-pyridyl) methyl] -1H-pyrrole-2-carboxylate (BCX-180, 9.5 g, 23.25 mmol) are added in a 300 ml glass lined glass pressure vessel to 90 ml of ammonium methylate (obtained by passing ammonia into anhydrous methanol at a temperature of 0-5 ^o C to saturation) and the reaction vessel is sealed. This vessel is heated to a temperature of 85-90 ^o C for 12 hours and then cooled to room temperature. Volatiles were evaporated using a rotary evaporator and 150 ml of 1N HCl and 100 ml of chloroform were added to the resulting residue. The resulting mixture was vigorously stirred and the HCl extract was isolated. The HCl extract was neutralized using an aqueous solution of sodium carbonate. The precipitated solid was isolated by filtration, dried in air and extracted with 2 L of ethanol in a Soxhlet apparatus for 48 hours. Ethanol was evaporated and the resulting light brown solid residue (3.1 g) was stirred with 20 ml of deionized (DI) water. Concentrated (37%) hydrochloric acid was added dropwise until all of the solid had dissolved (2.1 ml, 25.7 mmol). The clear solution was treated with activated carbon (700 mg) at a temperature of 50 ^° C. for 5 minutes and filtered through a short column filled with zeolite. The obtained filtrate was concentrated on a rotary evaporator at a temperature of 50-55 ^° C. to obtain a white solid, first dried in air and then in vacuum at a toluene distillation temperature. The mass of the obtained white solid is 3.6 g (48%), so pl. 340-342 ^o C.

Analysis: calculated for C ₁₂ H ₁₁ N ₅ O • 2HCl • 0.5H ₂ O: C 44.59; H 4.67; N, 21.67; Found: C, 44.59; H 4.57; N, 21.47.

 Example 69. Prepare a pharmaceutical composition for intraperitoneal injection for testing substances (IIA). For this, an intraperitoneal injectable solution containing the substance of Example 64 is dissolved in an aqueous vehicle that contains 10% DMSO.

Example 70. Intraperitoneally injected into Lewis Rats the experimental composition of Example 69 containing the substance (IIA) at a dose of 30 mg of the substance (IIA) by injection twice a day. Use control groups that receive only filler. After a certain time after administration, animals are euthanized and plasma samples are obtained. Plasma is extracted with cold 0.5N HClO ₄ and neutralized with solid NH ₄ HCO ₃ . After removal of the perchlorate salts, the extract is subjected to HPLC on a reversible phase column (Spherisorb ODSI). A significant increase in plasma inosine was observed in plasma taken from animals receiving substance (IIA).

 Examples 71-73. Substances such as in examples 66-68, were used to obtain pharmaceutical compositions in accordance with the method of example 69 and the resulting injectable solutions were tested in accordance with the method of example 70. A significant increase in plasma inosine was observed in plasma taken from animals receiving the substance of the invention.

Example 74. 1-Cyclohexenylacetonitrile is used in the syntheses to obtain the compounds of the invention. In dry N ₂ atmosphere, a solution of 1-cyclohexenylacetonitrile (9.2 g, 75.92 mmol) in anhydrous tetrahydrofuran (THF, 10 ml) was added to a stirred mixture of sodium hydride (3.18 g, 132.86 mmol) and formic ethyl ester acid (30.14 g, 406.93 mmol) in 50 ml of THF; the resulting mixture was stirred at room temperature for about 18 hours. The volatile material was evaporated in vacuo at room temperature. Water (30 ml) was added to the residue at 0 ^° C., the solution was adjusted to pH 6 by the addition of 6N HCl dropwise. The resulting precipitate as a heavy oil is extracted with CHCl ₃ . The extract was washed with water and dried with Na ₂ SO ₄ , the resulting organic layer was evaporated to give the crude formyl derivative as a red-brown oil (9.6 g).

Example 75. Glycine methyl ester hydrochloride (16.68 g, 132.85 mmol) and anhydrous sodium acetate (10.89 g, 132.85 mmol) were added to the solution of the crude formyl derivative (9.6 g) of Example 74 without further purification in a mixture of MeOH / H ₂ O (4: 1,150 ml). After 24 hours, MeOH was evaporated in vacuo and the mixture of water and oil was extracted with CHCl ₃ . The CHCl ₃ layer was dried (Na ₂ SO ₄ ) and evaporated to give an amber oil, which was fed to a silica gel column. Elution with CHCl ₃ afforded the desired enamine: yield 4.5 g.

Example 76. In a nitrogen atmosphere, ethyl chloroformate (3.04 g, 28.06 mmol) was added dropwise to the enamine solution of example 75 (4.12 g, 18.70 mol) and 1,5-diazabicyclo [4.3.0 ] non-5-ena (DBN, 6.96 g, 56.11 mmol) in dry CH ₂ Cl ₂ (100 ml) with external cooling in an ice bath. After stirring at 0 ^° C. for 1 h, the solution was allowed to stand at room temperature overnight. After checking the progress of the reaction with TCX, an additional amount of ClCO ₂ Et (0.5 ml) and DBN (3.0 ml) are added to complete the reaction, then the solution is left for 24 hours. The volatile material is evaporated in vacuo, the viscous residue is purified on a short column silica gel (the main purpose of which is to remove a less mobile DBN), obtaining N-protected pyrrole, which is used for the next step without further purification.

Example 77. To a solution of the N-protected pyrrole of Example 76 (3.0 g, 10.26 mmol) in MeOH (100 ml) was added solid Na ₂ CO ₃ (2.71 g, 25.65 mmol) and the reaction mixture was stirred at room temperature for 48 h with separation of the resulting unlocked pyrrole. The mixture was evaporated to dryness, the residue was triturated with water (50 ml) to dissolve the inorganic substances and extracted with CHCl ₃ (3 x 50 ml). The extract was dried (Na ₂ SO ₄ ) and evaporated to give a viscous resin, which was purified on a silica gel column using CHCl ₃ as an eluent; yield 2.04 g; so pl. 125 ^o C.

Example 78. Benzoylisothiocyanate (0.76 g, 4.65 mmol) was added dropwise to a solution of the unblocked pyrrole of Example 77 (0.91 g, 4.13 mmol) in dry CH ₂ Cl ₂ (30 ml). After 1 h at room temperature, the solution was evaporated and the gummy residue was triturated with methanol to obtain a thioureide derivative; yield 0.70 g, mp 170 ^o C.

Example 79. Methyl iodide (0.678 g, 4.78 mmol) was added to a solution of the thioureide derivative of Example 78 (0.630 g, 1.64 mmol) and DBN (0.230 g, 1.85 mmol) in dry CH ₂ Cl ₂ (50 ml) at 0 ^° C. The solution was stirred at 0 ^° C. for 15 minutes, at ambient temperature for 1 hour, and then evaporated in vacuo. A solution of the residue in CHCl _{3 was} chromatographed on a silica gel column using CHCl ₃ as an eluent to obtain homogeneous fractions of the methylthio compound as an intermediate; yield 0.7 g.

 Example 80

Раствор метилтиосоединения примера 79 (0,70 г, 1,76 ммоль) в 50 мл MeOH, который был насыщен NH₃ при 0^oC, нагревают при 90-95^oC в течение 24 ч в футерованной стеклянными плитками бомбе из нержавеющей стали. Содержимое охлажденной бомбы выпаривают в вакууме, получая смесь вещества (IIIA), бензамида и побочного продукта, который является 2-метилтиопроизводным, в отличие от 2-аминосоединения (IIIA). Смесь энергично перемешивают в течение нескольких минут с приблизительно 50 мл Et₂O, нерастворимое белое твердое вещество отфильтровывают и промывают с помощью Et₂O. Фильтрат содержал большую часть бензамида и 2-метилтиопроизводного. Раствор нерастворимого в Et₂O твердого вещества (0,342 г) в MeOH выпаривают с приблизительно 10 г силикагеля. Измельченный в порошок остаток наносят равномерным слоем на верхнюю часть колонны с силикагелем, который затем элюируют с помощью смеси CHCl₃/MeOH/HOAc (95:5:1), получая 2-метилтиопроизводное как побочный продукт и желаемое 2-аминосоединение (IIIA). Соединение (IIIA) перекристаллизовывают экстракцией в кипящем изопропилацетате в аппарате Сокслета. Белые кристаллы собирают в три массы и высушивают в вакууме над P₂O₅ при 110^oC в течение 7 ч; выход 44% т.пл. 280^oC (разл.).

A solution of the methylthio compound of Example 79 (0.70 g, 1.76 mmol) in 50 ml of MeOH that was saturated with NH ₃ at 0 ^° C was heated at 90-95 ^° C for 24 hours in a glass-lined stainless steel bomb. The contents of the chilled bomb are evaporated in vacuo to give a mixture of a substance (IIIA), benzamide and a by-product that is 2-methylthio derivative, as opposed to the 2-amino compound (IIIA). The mixture was vigorously stirred for several minutes with approximately 50 ml Et ₂ O, the insoluble white solid was filtered off and washed with Et ₂ O. The filtrate contained most of the benzamide and 2-methylthio derivative. A solution of Et ₂ O insoluble solid (0.342 g) in MeOH was evaporated with approximately 10 g of silica gel. The powder residue was uniformly applied to the top of the silica gel column, which was then eluted with CHCl ₃ / MeOH / HOAc (95: 5: 1) to give the 2-methylthio derivative as a by-product and the desired 2-amino compound (IIIA). Compound (IIIA) was recrystallized by extraction in boiling isopropyl acetate in a Soxhlet apparatus. White crystals are collected in three masses and dried in vacuum over P ₂ O ₅ at 110 ^o C for 7 hours; yield 44% mp 280 ^o C (decomp.).

Analysis for C ₁₂ H ₁₄ N ₄ O • 0.6H ₂ O: Calculated: C 59.78; H 6.35; N, 23.23; Found: C, 59.98; H 6.46; N, 23.15.

Example 81. The substance of example 80 was tested for enzymatic inhibitory activity. An enzymatic analysis of purine nucleoside phosphorylase (PNP) is carried out, in which the PNP activity of a substance is determined by radiochemical measurement of the formation of [ ¹⁴ C] -poxanthine from [ ¹⁴ C] -inosine (Biomedicina, 33, 39, 1980), using the spleen of the lower leg as a source of enzyme . At 1 mM phosphate, the IC ₅₀ is 1.9 μM; at 50 mM phosphate, the IC ₅₀ is 19 μM.

 Example 82. Following the methods described in examples 74-80, substances (IIIB) and (IIIC) were prepared using 2- and 3-cyclohexenylacetonitrile, respectively, as starting materials. The substances were tested as in Example 81, and significant enzymatic inhibitory activity was observed.

Examples 83-87. The following substances of the invention are obtained: 2-amino-7- (R) -3H, 5H-pyrrolo [3,2-d] pyrimidin-4-ones, in which the R group is as follows:
Example 83: R 3-methyl-2-cyclohexenyl;
Example 84: R 2-chloro-3-cyclohexenyl;
Example 85: R 3-trifluoromethyl-1-cyclohexenyl;
Example 86: R 3-methoxy-1-cyclohexenyl;
Example 87: R 2-fluoro-3-cyclohexenyl.

 Substances obtained by following the methods described in examples 74-81, using the corresponding cyclohexenylacetonitriles as starting materials.

 Example 88. Prepare a pharmaceutical composition for intraperitoneal administration for testing the substance (IIIA). The intraperitoneal injection solution containing the substance (IIIA) is dissolved in an aqueous vehicle that contains 10% DMSO.

Example 89. Intraperitoneally injected into Lewis Rats an experimental composition containing the substance (IIIA) of example 88, at a dose of 30 mg of the substance (IIIA) with injection, twice a day. Use control groups that receive only filler. After a certain time after administration, animals are euthanized and plasma samples are obtained. Plasma is extracted with cold 0.5N HClO ₄ and neutralized with solid NH ₄ HCO ₃ . After removal of the perchlorate salts, the extract is subjected to HPLC on a reversible phase column (Spherisorb ODSI). A significant increase in plasma inosine is observed in plasma taken from animals receiving the substance (IIIA).

 Examples 90-94. The substances obtained as in examples 83-87 were used to prepare drugs in accordance with the method of preparation of example 88 and the injectable solutions obtained were tested in accordance with the method of example 89. A significant increase in plasma inosine was observed in plasma taken from animals receiving substances inventions.

 Example 95

Вещество (IIID) получают, используя 2-амино-7-(1-циклогексенил)-3H,5H-пирроло[3,2-d] пиримидин-4-он в качестве промежуточного соединения. Раствор промежуточного соединения (0,2 г, 0,86 ммоль) в этаноле (50 мл) гидрируют с помощью 10%-ного Pd-C (50 мг) при давлении 45 фунтов/дюйм² в течение 16 ч и горячим фильтруют через цеолит. Фильтрат выпаривают досуха, остаток кристаллизуют из горячего этанола, получая вещество (IIID);
выход 157 мг (78%); т.пл. более 300^oC (разл.).

The substance (IIID) was prepared using 2-amino-7- (1-cyclohexenyl) -3H, 5H-pyrrolo [3,2-d] pyrimidin-4-one as an intermediate. A solution of intermediate (0.2 g, 0.86 mmol) in ethanol (50 ml) was hydrogenated using 10% Pd-C (50 mg) at 45 lb / ⁱⁿ² for 16 h and hot filtered through Celite . The filtrate was evaporated to dryness, the residue was crystallized from hot ethanol to give the substance (IIID);
yield 157 mg (78%); so pl. more than 300 ^o C (decomp.).

Analysis for C ₁₂ H ₁₆ N ₄ O • 0 • EtOH: Calculated: C 61.80; H 7.10; N, 23.51; Found: C, 61.95; H 7.43; N, 23.56.

Example 96. The substance (IIID) obtained in Example 95 was tested for enzyme-inhibitory activity as in Example 81. At 1 mM phosphate, IC ₅₀ is 1.3 μM, and at 50 mM phosphate, IC ₅₀ is 145 μM.

Example 97. 3- (2-Adamantyl) propionitrile was prepared in this example using the methodology of M. Ohno et al. J. Org. Chem. 53, 1285 (1988). A solution of 2-bromo-adamantane (20 g, 92.96 mmol), Bu ₃ SnH (32.46 g, 111.5 mol), acrylonitrile (9.86 g, 185.92 mmol) and AIBN (740 mg) in toluene ( 280 ml) was stirred at reflux for 3 hours. The reaction mixture was washed with ammonia water (0.4 M, 500 ml), the organic layer was washed with water, dried over MgSO ₄ and evaporated. The residue is distilled at a temperature between 110-118 ^o C and about 0.2 mmHg; the fractions are combined to give a crude sample of contaminated 3- (2-adamantyl) propionitrile with tin complexes, which is purified on a silica gel column with hexanes, followed by hexane / ethyl acetate 97: 3 and hexane / ethyl acetate 95: 5; yield 9.4 g (53.4%), mp. semi-solid.

Example 98. The 3- (2-Adamantyl) propionitrile of Example 97 is further used to synthesize the compounds of the invention. In a dry N ₂ atmosphere, a mixture of 3- (2-adamantyl) propionitrile (7.0 g, 36.99 mmol), sodium hydride (1.7 g, 73.95 mmol) and anhydrous tetrahydrofuran (75 ml) is heated at 52 ^o C in a water bath for 15 minutes, a solution of formic acid ethyl ester (13.69 g, 184.89 mmol) in THF (100 ml) was added over 45 minutes. After 2 hours at 50-55 ^° C., a second portion of NaH (0.8 g) and HCO ₂ Et (7.5 ml) were added and the reaction mixture was stirred for about 2 days. A third portion of HCO ₂ Et (7.5 ml) and NaH (0.8 g) were added and left at room temperature for about 24 hours (unreacted nitrile is inert in the next step and can be regenerated in the first purification step). Steep paste is stirred overnight and allowed to cool to room temperature. The volatile material was evaporated under reduced pressure, the remaining pale yellow peel was dissolved in a minimum volume of cold water (approximately 150 ml) at 0 ^° C. The solution was adjusted to pH 6.0 by adding 6N HCl and extracted with CHCl ₃ (3 x 100 ml) . The extract was washed with water, dried over Na ₂ SO ₄ and evaporated in vacuo to a thick amber oil. This crude product is used in the next reaction without further purification.

Example 99. Glycine methyl ester hydrochloride (6.96 g, 55.46 mmol) and anhydrous sodium acetate (4.55 g, 55.46 mmol) were added to a solution of crude formyl derivative (8.0 g) in MeOH / H ₂ O (4: 1,500 ml). After 24 hours, MeOH was evaporated in vacuo and the mixture of water and oil was extracted with CHCl ₃ . The CHCl ₃ layer was dried (Na ₂ SO ₄ ) and evaporated to give an amber oil, which was fed to a silica gel column. Elution with CHCl ₃ gives two main layers: 3- (2-adamantyl) propionitrile (used as starting material in the previous step) and the desired enamine; yield 6 g.

Example 100. In a nitrogen atmosphere, ethyl chloroformate (2.82 g, 26.0 mmol) was added dropwise to the enamine solution of Example 99 (5.0 g, 17.34 mmol) and 1,5-diazabicyclo [4.3.0 ] non-5-ena ("DBN", 6.46 g, 52.0 mmol) in dry CH ₂ Cl ₂ (50 ml) with external cooling in an ice bath. After stirring at 0 ^° C. for 1 h, the solution was allowed to stand at room temperature overnight. After monitoring the progress of the reaction by TCX, additional ClCO ₂ Et (1.81 ml) and DBN (3.23 g) were added to complete the reaction and the solution was left to stand for 24 hours. The volatile material was evaporated in vacuo, the viscous residue was purified on a short column silica gel (the main purpose of which is the removal of less mobile DBN), obtaining N-protected pyrrole, which is used for the next step without further purification.

Example 101. To a solution of the crude N-protected pyrrole of Example 100 (6.0 g, 16.59 mmol) in MeOH (100 ml) was added solid Na ₂ CO ₃ (4.39 g, 41.49 mmol), the reaction mixture was stirred at room temperature for 48 h with separation of the resulting unlocked pyrrole. The mixture is evaporated to dryness, the residue is thoroughly triturated with water into a powder (50 ml) to dissolve inorganic substances and extracted with CHCl ₃ (3 • 100 ml). The extract was dried (Na ₂ SO ₄ ) and evaporated to give a viscous resin, which was crystallized with ether; yield 4 g; so pl. 162-163 ^o C.

Example 102. Benzoylisothiocyanate (1.22 g, 7.47 mmol) was added dropwise to a solution of the unblocked pyrrole of Example 101 (1.91 g, 6.62 mmol) in dry CH ₂ Cl ₂ (50 ml). After 1 h at room temperature, the solution was evaporated, the gummy residue was dissolved in Et ₂ O (100 ml) with an almost immediate release of crystalline solid, which was filtered off. The mother liquor Et ₂ O is heated to boiling and diluted with an equal volume of hot cyclohexane. With slow cooling, the solution gives an additional amount of thioureid derivative; yield 2.81 g (95%); so pl. 193-194 ^o C.

Example 103. Methyl iodide (2.46 g, 17.39 mmol) was added to a solution of the thioureide derivative of Example 102 (2.7 g, 5.98 mmol) and DBN (0.82 g, 6.57 mmol) in dry CH ₂ Cl ₂ (50 ml) at 0 ^° C. The solution was stirred at 0 ^° C. for 15 minutes, at ambient temperature for 1 hour, and then evaporated in vacuo to give a crude sample of the methylthio compound as an intermediate; yield 2.78 g (crude).

 Example 104

Раствор метилтиосоединения примера 103 (2,78 г, 5,18 ммоль) в 150 мл MeOH, который был насыщен NH₃ при 0^oС, нагревают при 90-95^oC в течение 24 ч в футерованной стеклянными плитками бомбе из нержавеющей стали. Содержимое охлажденной бомбы выпаривают в вакууме, получая смесь вещества (IVA), бензамида и побочного продукта, который является 2-метилтиопроизводным в отличие от 2-аминосоединения (IVA). Смесь энергично перемешивают в течение нескольких минут с приблизительно 75 мл Et₂O и нерастворимое белое твердое вещество отфильтровывают с помощью Et₂O. Фильтрат содержал большую часть бензамида и 2-метилтиопроизводное. Раствор нерастворимого в Et₂O твердого вещества (1,38 г) в MeOH выпаривают с приблизительно 25 г силикагеля. Измельченный в порошок остаток наносится равномерно на верхнюю часть колонки с силикагелем, которую затем элюируют с помощью CHCl₃/MeOH/HOAc (95:5:1), получая 2-метилтиопроизводное как побочный продукт и желаемое 2-аминосоединение (IVA). (IVA) продукт перекристаллизовывают при экстракции в кипящем изопропилацетате в аппарате Сокслета. Белые кристаллы собирают в три массы и высушивают в вакууме над P₂O₅ при 110^oC в течение 7 ч; выход 51,5% т.пл. более 350^oC (разл.).

A solution of the methylthio compound of Example 103 (2.78 g, 5.18 mmol) in 150 ml of MeOH that was saturated with NH ₃ at 0 ^° C was heated at 90-95 ^° C for 24 hours in a glass-lined stainless steel bomb. The contents of the cooled bomb are evaporated in vacuo to give a mixture of a substance (IVA), benzamide and a by-product that is 2-methylthio derivative in contrast to the 2-amino compound (IVA). The mixture was vigorously stirred for several minutes with approximately 75 ml Et ₂ O and the insoluble white solid was filtered off with Et ₂ O. The filtrate contained most of the benzamide and 2-methylthio derivative. A solution of Et ₂ O insoluble solid (1.38 g) in MeOH was evaporated with approximately 25 g of silica gel. The powdery residue is applied evenly to the top of the silica gel column, which is then eluted with CHCl ₃ / MeOH / HOAc (95: 5: 1) to give the 2-methylthio derivative as a by-product and the desired 2-amino compound (IVA). (IVA) the product is recrystallized by extraction in boiling isopropyl acetate in a Soxhlet apparatus. White crystals are collected in three masses and dried in vacuum over P ₂ O ₅ at 110 ^o C for 7 hours; yield 51.5% mp more than 350 ^o C (decomp.).

Analysis for C ₁₇ H ₂₂ N ₄ O • 0.21MeOH • 0.22H ₂ O: Calculated: C 66.88; H 7.59; N, 18.12; Found: C, 66.86; H 7.59; N, 18.12.

Example 105. The substance of example 104 was tested for enzymatic inhibitory activity. For this, an enzymatic analysis of purine nucleoside phosphorylase (PNP) is carried out, in which the PNP activity (IC ₅₀ ) is determined for the substance by radiochemical measurement of the formation of [ ¹⁴ C] -poxanthine from [ ¹⁴ C] -inosine (Biomedicine, 33, 39, 1980) using a calf spleen as a source of enzyme. At 1 mM phosphate, the IC ₅₀ is 0.090 μM and at 50 mM phosphate, the IC ₅₀ is 2.5 μM.

Examples 106-110. The following substances of the invention are obtained: 2-amino-7- (CH ₂ R) -3H, 5H-pyrrolo [3,2-d] pyrimidin-4-ones, in which R is a substituted 2-adamantyl group:
Example 106: R 2- (1-methyl) -adamantyl;
Example 107: R 2- (1-chloro) -adamantyl;
Example 108: R 2- (1-trifluoromethyl) adamantyl;
Example 109: R 2- (1-methoxy) -adamantyl;
Example 110: R 2- (1-fluoro) -adamantyl.

 Substances are prepared following the methods described in Examples 98-104 using the corresponding 3- (2-adamantyl) propionitriles as starting materials.

Examples 111-116. The following substances of the invention are obtained: 2-amino-7- (CH ₂ R) -3H, 5H-pyrrolo [3,2-d] pyrimidin-4-ones, in which R is the following 1-adamantyl group:
Example 111: R 1- (2-methyl) -adamantyl;
Example 112: R 1- (2-chloro) -adamantyl;
Example 113: R 1- (2-trifluoromethyl) adamantyl;
Example 114: R 1- (2-methoxy) -adamantyl;
Example 115: R 1- (2-fluoro) -adamantyl;
Example 116: R 1-Adamantyl.

 Substances are prepared following the methods described in Examples 98-104 using the corresponding 3- (1-adamantyl) propionitriles as starting materials.

 Example 117. Prepare a pharmaceutical composition for intraperitoneal injection for testing substances (IVA). An intraperitoneal injection solution containing a substance (IVA) is dissolved in an aqueous vehicle that contains 10% DMSO.

Example 118. An experimental composition was administered intraperitoneally in Lewis Rats containing the substance (IVA) of Example 117 at a dose of 30 mg of the substance (IVA) by injection twice a day. Use control groups that receive only filler. After a certain time after administration, animals are euthanized and plasma samples are obtained. Plasma is extracted with cold 0.5N HClO ₄ and neutralized with solid NH ₄ HCO ₃ . After removal of the perchlorate salts, the extract is subjected to HPLC on a reversible phase column (Spherisorb ODSI). A significant increase in plasma inosine is observed in plasma taken from animals receiving the substance (IVA).

 Examples 119-129. The substances obtained as in examples 106-116 were used to prepare drugs in accordance with the method of preparation of example 117, the injectable solutions obtained were tested in accordance with the method of example 118. An increase in plasma inosine in plasma taken from animals receiving the compounds of the invention was observed.

Example 130. In this example, 3-cyclopentylpropionitrile is obtained. 3-Cyclopentylpropionitrile chloride (57.7 g, 0.36 mol) was added dropwise to a large excess of concentrated ammonium hydroxide (400 ml), cooled in an ice / salt bath. A heavy suspension of a white solid is stirred overnight, collected by filtration, washed with cold water and recrystallized from about 2 L of boiling water. The shiny white amide plates are dried in vacuo over P ₂ O ₅ ; yield 31.6 g (62.3%), mp. 122 ^o C.

With protection from atmospheric moisture, a solution of amide (23.5 g, 0.166 mol) in POCl ₃ (150 ml) was heated at 120 ^° C for 1 h. The oil bath was cooled to about 70 ^° C, the excess POCl _{3 was} distilled off in vacuo, cooled the residue is poured onto ice (about 300 g). The mixture was neutralized by careful addition of solid Na ₂ CO ₃ and extracted with several portions of Et ₂ O. The dried extract over Na ₂ SO _{4 was} evaporated to give a clear pale yellow oil, which was distilled in vacuo to give the desired nitrile; yield 16.86 g (82%), mp 88.0-88.5 ^o C / 8.7 mm. MS (EI): m / z 122 (MH) ⁺ ; IR (sar. Film), 2245 cm ^-1 (CN); ¹ H NMR, δ 1.67 (q, 2, CH ₂ CH ₂ CN), 2.36 (t, 2, -CH ₂ CN), complex multiplets centered around 1.11, 1.63, 1.86 (cyclopentyl protons).

Example 131. The 3-cyclopentylpropionitrile of the previous example is further processed in syntheses of the invention. In an atmosphere of dry N _{2, a} mixture of 3-cyclopentylpropionitrile (14.8 g, 0.12 mol), sodium hydride (5.8 g, 0.24 mol) and anhydrous tetrahydrofuran (300 ml) is heated at 52 ^o C in a water bath within 15 minutes, a solution of formic acid ethyl ester (13.3 g, 0.18 mol) in THF (100 ml) was added over 45 minutes. After 2 hours at 50-55 ^° C., a second portion of NaH (1.9 g) and HCO ₂ Et (5.0 ml) were added, followed by a third portion of HCO ₂ Et over 30 minutes (unreacted nitrile is inert in the next step and can be regenerated in the first purification step). The steep paste is stirred overnight and allowed to cool to room temperature. The volatile material was evaporated under reduced pressure, the remaining pale yellow peel was dissolved in a minimum volume of cold water (about 150 ml) at 0 ^° C. The solution was adjusted to pH 6.0 with 6N HCl and extracted with CHCl ₃ (3 x 100 ml). The extract was washed with water, dried over Na ₂ SO ₄ and evaporated in vacuo to a thick amber oil. This crude product (15.6 g) was used in the next reaction without further purification.

Example 132. Glycine methyl ester hydrochloride (19.31 g, 0.154 mol) and anhydrous sodium acetate (12.61 g, 0.154 mol) were added to a solution of the crude formyl derivative of the previous example (15.6 g) in MeOH / H ₂ O ( 4: 1, 500 ml). After 24 hours, MeOH was evaporated in vacuo, the mixture of water and oil was extracted with CHCl ₃ . The CHCl ₃ layer was dried (Na ₂ SO ₄ ) and evaporated to give an amber oil, which was fed to a silica gel column. Elution with CHCl ₃ gave two main layers: 3-cyclopentylpropionitrile (8.22 g, 66.7 mmol or 55.6% nitrile, used as starting material in the previous step); desired enamine (3.45 g, 29.1% of theoretical yield, adjusted for the amount of nitrile present in the starting material; MS (FAB): 223 (M + H) ⁺ ).

Example 133. In a nitrogen atmosphere, ethyl chloroformate (2.53 g, 23.3 mmol) was added dropwise to the enamine solution of the previous example (3.45 g, 15.5 mmol) and DBN (5.78 g, 46, 6 mmol) in dry CH ₂ Cl ₂ (50 ml) with external cooling in an ice bath. After stirring at 0 ^° for 1 hour, the solution was kept at room temperature overnight. After checking the progress of the reaction by TCX, an additional amount of ClCO ₂ Et (0.5 ml) and DBN (3.0 ml) were added to complete the reaction, the solution was kept for 24 h. The volatile substance was evaporated in vacuo, the viscous residue was purified on a short column with silica gel (the main purpose of which is the removal of less mobile DBN), obtaining N-protected pyrrole (4.50 g, 98%), which is used for the next step without further purification.

Example 134. To a solution of the N-protected pyrrole of the previous example (4.50 g, 15.3 mmol) in MeOH (100 ml) was added solid Na ₂ CO ₃ (1.62 g, 15.3 mmol) and the reaction mixture was stirred at room temperature for 48 h with separation of the resulting unlocked pyrrole. The mixture was evaporated to dryness, the residue was triturated with H ₂ O (50 ml) to dissolve the inorganic substances and extracted with CHCl ₃ (3 x 100 ml). The extract was dried (Na ₂ SO ₄ ) and evaporated to give a viscous resin, which crystallized upon drying in vacuo; yield 2.97 g (87.4%) of a substance suitable for use as an intermediate without further purification. Deeper purification can be carried out by column chromatography using silica gel / CHCl ₃ or recrystallization from a mixture of toluene / cyclohexane (1: 3).

Example 135. Benzoylisothiocyanate (2.62 g, 16.03 mmol) was added dropwise to a solution of the unblocked pyrrole of Example 134 (2.97 g, 13.36 mmol) in dry CH ₂ Cl ₂ (100 ml). After 1 h at room temperature, the solution was evaporated and the gummy residue was dissolved in Et ₂ O (100 ml) with an almost immediate separation of the crystalline solid; yield 1.75 g. The filtrate Et ₂ O was heated to boiling and diluted with an equal volume of hot cyclohexane. With slow cooling, the solution gives an additional 1.58 g of thioureid derivative; total yield 3.33 g (64.6%). A small amount of the thioureido compound was recrystallized from a hot mixture of Et ₂ O / cyclohexane (15 ml each); so pl. 123-125 ^° C. MS (FAB): 386 (M + H) ⁺ .

Analysis for C ₂₀ H ₂₃ N ₃ O ₃ S • 0.45C ₆ H ₁₂ Calcd: C 64.40; H 6.76; N, 9.93; Found: C, 64.51; H 7.10; N, 9.93.

Example 136. Methyl iodide (2.60 g, 18.32 mmol) was added to a solution of the thioureido compound of Example 135 (3.21 g, 8.33 mmol) and DBN (1.24 g, 9.99 mmol) in dry CH ₂ Cl ₂ (80 ml) at 0 ^° C. The solution was stirred at 0 ^° C. for 15 minutes at ambient temperature for 1 hour, and then evaporated in vacuo. A solution of the residue in CHCl _{3 was} chromatographed on a silica gel column using CHCl ₃ as an elution solvent to obtain homogeneous fractions of the methylthio compound as an intermediate; yield 2.46 g (74%).

 Example 137

Раствор метилтиосоединения примера 136 (2,07 г, 5,18 ммоль) в 150 мл MeOH, который был насыщен NH₃ при 0^oC, нагревают при 90-95^oC в течение 24 ч в футерованной стеклянными плитками бомбе из нержавеющей стали. Содержимое охлажденной бомбы выпаривают в вакууме, получая смесь вещества (IVC), бензамида и побочного продукта, который является 2-метилтиопроизводным в отличие от 2-аминосоединения (IVC). Смесь энергично перемешивают в течение нескольких минут с приблизительно 75 мл Et₂O и нерастворимое белое вещество отфильтровывают и промывают Et₂O. Фильтрат содержал большую часть бензамида и 2-метилтиосоединения. Раствор нерастворимого в Et₂O твердого вещества (1,13 г) в MeOH выпаривают с приблизительно 10 г силикагеля. Измельченный в порошок остаток наносят равномерно на верхнюю часть колонки с силикагелем, которую затем элюируют с помощью смеси CHCl₃/MeOH/HOAc (95:5:1), получая 2-метилтиопроизводное как побочный продукт (252 мг, MS (FAB): 264 (M+H)⁺) и желаемое 2-аминосоединение (IVC) (679 мг, 56,4%). (IVC) перекристаллизовывают при экстракции в кипящем изопропилацетате в аппарате Сокслета. Белые кристаллы собирают в три порции и высушивают в вакууме над P₂O₅ при 110^oC в течение 7 ч; выход 540 мг (44,9%); т.пл. 324-326^oC (разл.); MS (FAB): 233 (M+H)⁺.

A solution of the methylthio compound of Example 136 (2.07 g, 5.18 mmol) in 150 ml of MeOH that was saturated with NH ₃ at 0 ^° C was heated at 90-95 ^° C for 24 hours in a glass-lined stainless steel bomb. The contents of the chilled bomb are evaporated in vacuo to give a mixture of a substance (IVC), benzamide and a by-product that is 2-methylthio derivative in contrast to the 2-amino compound (IVC). The mixture was vigorously stirred for several minutes with approximately 75 ml of Et ₂ O and the insoluble white material was filtered off and washed with Et ₂ O. The filtrate contained most of the benzamide and 2-methylthio compound. A solution of Et ₂ O insoluble solid (1.13 g) in MeOH was evaporated with approximately 10 g of silica gel. The powdered residue was applied evenly to the top of the silica gel column, which was then eluted with a mixture of CHCl ₃ / MeOH / HOAc (95: 5: 1) to give the 2-methylthio derivative as a by-product (252 mg, MS (FAB): 264 (M + H) ⁺ ) and the desired 2-amino compound (IVC) (679 mg, 56.4%). (IVC) is recrystallized by extraction in boiling isopropyl acetate in a Soxhlet apparatus. The white crystals are collected in three portions and dried in vacuo over P ₂ O ₅ at 110 ^° C. for 7 hours; yield 540 mg (44.9%); so pl. 324-326 ^° C (decomp.); MS (FAB): 233 (M + H) ⁺ .

Analysis for C ₁₂ H ₁₆ N ₄ O: Calculated: C 62.05; H 6.94; N, 24.12; Found: C, 62.04; H 7.11; N, 24.48.

Example 138. The substance of example 137 was tested for enzyme-inhibitory activity in accordance with the method of example 9. At 1 mM phosphate, IC _{50 was} 0.029 μM and at 50 mM phosphate, IC _{50 was} 1.8 μM.

Examples 139-142. The following substances of the invention are obtained: 2-amino-7- (CH ₂ R) -3H, 5H-pyrrolo [3,2-d] pyrimidin-4-ones, in which R is substituted cyclopentyl:
Example 139: R 3-methylcyclopentyl;
Example 140: R 2-chlorocyclopentyl;
Example 141: R 3-trifluoromethylcyclopentyl;
Example 142: R 3-methoxycyclopentyl.

 Substances are prepared following the methods described in Examples 130-137 using the corresponding 3- (substituted cyclopentyl) propionitriles as starting materials.

 Example 143. A pharmaceutical composition is prepared for intraperitoneal injection for testing a substance (IVC). An intraperitoneal injection solution containing a substance (IVC) is dissolved in an aqueous vehicle that contains 10% DMSO.

 Example 144. Using the method of Example 112, an experimental composition containing the substance (IVC) of Example 143 is administered intraperitoneally in Lewis Rats, and the results are compared with the controls. A significant increase in plasma inosine is observed in plasma taken from animals receiving the substance (IVC).

Example 145. Get 3-cyclohexylpropionitrile. A solution of cyclohexane propionic acid (50 g, 0.32 mol) and thionyl chloride (152 g, 1.28 mol) in 100 ml of benzene was kept overnight and then evaporated to an oily residue. The residue was added portionwise to a 28% aqueous ammonia solution (270 ml) at 25 ^° C. and the mixture was stirred for about 2 hours. The resulting product was collected by filtration, washed with cold water and recrystallized from about 2 L of boiling water. The shiny white amide plates are dried in vacuo over P ₂ O ₅ ; yield 45.5 g.

With protection from atmospheric moisture, a solution of the amide (45.5 g, 0.293 mol) in SOCl ₂ (200.3 g, 1.68 mol) is refluxed for about 6 hours. The oil bath is cooled to about 70 ^o C and excess SOCl _{2 was} distilled off in vacuo, the cooled residue was poured onto ice (about 300 g). The mixture was neutralized by careful addition of solid Na ₂ CO ₃ and extracted with several portions of Et ₂ O. The dried (Na ₂ SO ₄ ) extract was evaporated to give a clear, pale yellow oil, which was distilled in vacuo to give the desired nitrile; yield 42.0 g.

Example 146. The 3-cyclohexylpropionitrile of Example 145 is then used in the synthesis to prepare the compounds of the invention. In a dry N ₂ atmosphere, a mixture of 3-cyclohexylpropionitrile (22.3 g, 0.16 mol), sodium hydride (5.38 g, 0.224 mol) and anhydrous tetrahydrofuran (120 ml) is heated at 52 ^° C. in a water bath for 15 min., add a solution of formic acid ethyl ester (55.4 g, 0.75 mol) in THF (50 ml) over 45 minutes. After 2 hours at 50-55 ^° C., a second portion of NaH (2.0 g) and HCO ₂ Et (5.0 ml) are added (unreacted nitrile is inert in the next step and can be regenerated in the first purification step), the reaction mixture is stirred in for about 3 days at 55 ^o C and then allowed to cool to room temperature. The volatile material was evaporated under reduced pressure, the remaining pale yellow peel was dissolved in a minimum amount of cold water (about 75 ml) at 0 ^° C. The solution was adjusted to pH 6.0 by the addition of 6N HCl and extracted with CHCl ₃ (3 x 100 ml). The extract was washed with water, dried over Na ₂ SO ₄ and evaporated in vacuo to a thick amber oil. This crude product is used in the next reaction without further purification.

Example 147. Glycine methyl ester hydrochloride (30.60 g, 0.24 mol) and anhydrous sodium acetate (19.99 g, 0.24 mol) were added to a solution of the crude formyl derivative of the previous example (25.22 g) in a MeOH mixture / H ₂ O (4: 1,500 ml). After 24 hours, MeOH was evaporated in vacuo, the mixture of water and oil was extracted with CHCl ₃ . The CHCl ₃ layer was dried (Na ₂ SO ₄ ) and evaporated to give an amber oil, which was fed to a silica gel column. Elution with CHCl ₃ gives two basic layers: 3-cyclohexylpropionitrile (used as starting material in the previous step); desired enamine; yield 16 g.

Example 148. In a nitrogen atmosphere, ethyl chloroformate (1.38 g, 12.7 mmol) was added dropwise to the enamine solution of Example 147 (2.0 g, 8.46 mmol) and DBN (2.1 g, 16, 9 mmol) in dry CH ₂ Cl ₂ (50 ml) with external cooling in an ice bath. After stirring at 0 ^° C. for 1 h, the solution was kept at room temperature overnight. After checking the reaction progress by TCX, an additional amount of ClCO ₂ Et (0.5 ml) and DBN (1.5 ml) were added to complete the reaction, the solution was allowed to stand for 24 hours. The volatile material was evaporated in vacuo, the viscous residue was purified on a short column silica gel (the purpose of which is to remove a less mobile DBN), obtaining N-protected pyrrole, which is used for the next step without further purification.

Example 149. To a solution of the N-protected pyrrole of Example 148 (2.6 g, 8.43 mmol) in MeOH (100 ml), solid Na ₂ CO ₃ (2.23 g, 21.07 mmol) was added and the reaction mixture was stirred at room temperature for 48 h with separation of the resulting unlocked pyrrole. The mixture was evaporated to dryness, the residue was thoroughly triturated with water (50 ml) to dissolve inorganic substances and extracted with CHCl ₃ (3 x 100 ml). The extract was dried (Na ₂ SO ₄ ) and evaporated to give a viscous gum which was purified on a silica gel column using CHCl ₃ as an elution solvent; yield 1.67 g (84%), mp. 73-74 ^o C.

Example 150. Benzoylisothiocyanate (0.74 g, 4.02 mmol) was added dropwise to a solution of the unblocked pyrrole of Example 149 (0.95 g) in dry CH ₂ Cl ₂ (20 ml). After 1 h at room temperature, the solution was evaporated and the gummy residue was dissolved in Et ₂ O (100 ml) with an almost immediate separation of the crystalline solid. The mother liquor Et ₂ O is heated to boiling and diluted with an equal volume of hot cyclohexane. With slow cooling, the solution gives an additional thioureide derivative; yield 1.41 g (88%), mp. 156-157 ^o C.

Example 151. Methyl iodide (1.1 g, 7.6 mmol) was added to a solution of the thioureid derivative of Example 150 (0.96 g, 2.61 mmol) and 1,5-diazabicyclo [4.3.0] non-5-ene ( 0.38 g, 3.0 mmol) in dry CH ₂ Cl ₂ (20 ml) at 0 ^° C. The solution was stirred at 0 ^° C. for 15 minutes, at ambient temperature for 1 hour, and then evaporated in vacuo. A solution of the residue in CHCl _{3 was} chromatographed on a silica gel column with CHCl ₃ as an elution solvent, to obtain homogeneous fractions of the methylthio compound as an intermediate; yield 0.92 g.

 Example 152

Раствор метилтиосоединения примера 151 (0,8 г, 1,93 ммоль) в 50 мл MeOH, который был насыщен NH₃ при 0^oC, нагревают при 90-95^oC в течение 24 ч в футерованной стеклянными плитками бомбе из нержавеющей стали. Содержимое охлажденной бомбы выпаривают в вакууме, получая смесь вещества (IVD), бензамида и побочного продукта, который является 2-метилтиопроизводным, в отличие от 2-аминосоединения (IVD). Смесь энергично перемешивают в течение нескольких минут с приблизительно 75 мл Et₂, нерастворимое твердое белое вещество отфильтровывают и промывают с помощью Et₂O. Фильтрат содержал большую часть бензамида и 2-метилтиосоединения. Раствор нерастворимого в Et₂O твердого вещества (0,390 г) в MeOH выпаривают с приблизительно 10 г силикагеля. Измельченный в порошок остаток наносят равномерно на верхнюю часть колонки с силикагелем, которую затем элюируют смесью CHCl₃/MeOH/HOAc (95:5: 1), получая 2-метилтиопроизводное как побочный продукт и желаемое 2-аминосоединение (IVD). (IVD) перекристаллизовывают при экстракции в кипящем изопропилацетате в аппарате Сокслета. Белые кристаллы собирают в три порции и высушивают в вакууме над P₂O₅ при 110^oC в течение 7 ч; выход 49% т.пл. более 300^oC.

A solution of the methylthio compound of Example 151 (0.8 g, 1.93 mmol) in 50 ml of MeOH that was saturated with NH ₃ at 0 ^° C was heated at 90-95 ^° C for 24 hours in a glass-lined stainless steel bomb. The contents of the cooled bomb are evaporated in vacuo to give a mixture of a substance (IVD), benzamide and a by-product that is 2-methylthio derivative, as opposed to the 2-amino compound (IVD). The mixture was vigorously stirred for several minutes with approximately 75 ml of Et ₂ , the insoluble white solid was filtered off and washed with Et ₂ O. The filtrate contained most of the benzamide and 2-methylthio compound. A solution of Et ₂ O insoluble solid (0.390 g) in MeOH was evaporated with approximately 10 g of silica gel. The powder residue was applied evenly to the top of the silica gel column, which was then eluted with a mixture of CHCl ₃ / MeOH / HOAc (95: 5: 1) to give the 2-methylthio derivative as a by-product and the desired 2-amino compound (IVD). (IVD) is recrystallized by extraction in boiling isopropyl acetate in a Soxhlet apparatus. The white crystals are collected in three portions and dried in vacuo over P ₂ O ₅ at 110 ^° C. for 7 hours; yield 49% mp more than 300 ^o C.

Analysis for C ₁₃ H ₁₈ N ₄ O: Calculated: C 63.39; H 7.36; N, 22.74; Found: C, 63.50; H 7.74; N, 22.67.

Example 153. The substance of Example 152 was tested for enzymatic inhibitory activity as in Example 105. At 1 mM phosphate, IC _{50 was} 0.037 μM, and at 50 mM phosphate, IC _{50 was} 2.2 μM.

Example 154. The substance of Example 152 was tested to determine its effectiveness in inhibiting the toxicity of 2'-deoxyguanosine (d-GuO) (DA Schewach et al. Cancer es. 46, 519 (1986) and JCSirear et al. Agents and tions, 21, 253 (1987)). For this, CCRF-CEM cells are grown in RPMI-1640 medium. D-GuO was added to the suspended cultures of these cells at a stationary concentration (5.62 μM) and the substance at a variable concentration, the number of cells in a Coulter counter was determined 24, 48 and 72 hours after that. From these data, an IC ₅₀ of 2.0 μM was calculated as the concentration of the substance required to reduce cell growth between 0 and 72 hours by 50% of the cells of the control cultures.

 Example 155.

Получают 2-амино-7-(3-метилциклогексилметил)-3H, 5H-пирроло[3,2-d]пиримидин-4-он. Сначала используя способы, приведенные в примерах 146 152, но с 3-(3-метилбензил)-пропионитрилом в качестве исходного вещества, получают арильное производное 2-амино-7-(3-метилбензил)-3H, 5H-пирроло[3,2-d] пиримидин-4-он. Раствор арильного производного (0,2 г, 0,78 ммоль) в трифторуксусной кислоте (TFA) (20 мл) гидрируют с помощью PtO₂ при давлении 60 фунтов/дюйм в течение 24 ч. Катализатор отфильтровывают через цеолитный слой и фильтрат выпаривают. Остаток растирают в порошок с помощью метанола и оставляют в холодильнике на ночь. Полученная кристаллическая трифторацетатная соль осаждается из раствора и собирается фильтрованием. TFA-соль суспендируют в 8 мл воды, доводя до pH 8 конц. NH₄OH, измельчают ультразвуком. Чистый продукт собирают, промывают водой и сушат; выход 165 мг (81%), т.пл. 282^oC.

Get 2-amino-7- (3-methylcyclohexylmethyl) -3H, 5H-pyrrolo [3,2-d] pyrimidin-4-one. First, using the methods described in examples 146 to 152, but with 3- (3-methylbenzyl) propionitrile as the starting material, an aryl derivative of 2-amino-7- (3-methylbenzyl) -3H, 5H-pyrrolo is obtained [3,2 -d] pyrimidin-4-one. A solution of the aryl derivative (0.2 g, 0.78 mmol) in trifluoroacetic acid (TFA) (20 ml) was hydrogenated with PtO ₂ at a pressure of 60 psi for 24 hours. The catalyst was filtered through a zeolite layer and the filtrate was evaporated. The residue was triturated with methanol and left in the refrigerator overnight. The resulting crystalline trifluoroacetate salt precipitates from solution and is collected by filtration. The TFA salt is suspended in 8 ml of water, adjusted to pH 8 conc. NH ₄ OH, crushed by ultrasound. The pure product is collected, washed with water and dried; yield 165 mg (81%), mp. 282 ^o C.

Analysis for C ₁₄ H ₂₀ N ₄ O: Calculated: C 64.60; H 7.74; N, 21.52; Found: C, 64.24; H 7.96; N 21.51%
Example 156

Способ, описанный в примере 155, повторяют для получения 2-амино-7-(3-трифторметилциклогексилметил)-3H, 5H-пирроло [3,2-d] пиримидин-4-она, используя 3-(3-трифторметилбензил)-пропионитрил в качестве исходного вещества; выход 69% т.пл. 165^oC.

The method described in example 155 is repeated to obtain 2-amino-7- (3-trifluoromethylcyclohexylmethyl) -3H, 5H-pyrrolo [3,2-d] pyrimidin-4-one using 3- (3-trifluoromethylbenzyl) propionitrile as a starting material; yield 69% mp 165 ^o C.

Analysis for C ₁₄ H ₁₇ N ₄ OF ₃ • 0.6H ₂ O: Calculated: C 51.72; H 5.64; N, 17.23; Found: C, 51.82; H 5.71; N 16.81%
Example 157. The substance obtained in example 155 was tested for enzyme-inhibitory activity, as in example 105. At 1 mM phosphate, IC _{50 was} 0.025 μM, at 50 mM phosphate, IC _{50 was} 0.820 μM.

Example 158. The substance obtained in example 156 was tested for enzyme-inhibitory activity, as in example 105. At 1 mM phosphate, IC ₅₀ is 0.020 μM, at 50 mM phosphate, IC ₅₀ is 0.740 μM.

Example 159. In this example, 3-cycloheptylpropionitrile was prepared according to the method of Example 97 using a solution of 2-bromocycloheptane (25.57 g, 144.38 mmol), Bu ₃ SnH (50.42 g, 173.26 mmol), acrylonitrile (15.32 g, 288.77 mmol) and AIBN (1.13 g) in toluene (300 ml); yield 16 g as an oil.

Example 160. The 3-cycloheptyl propionitrile of Example 159 is then used in the synthesis to prepare the compounds of the invention. In a dry N ₂ atmosphere, a mixture of 3-cycloheptyl propionitrile (8.5 g, 56.19 mmol), sodium hydride (2.6 g, 112.39 mmol) is heated in a water bath for 15 min, a solution of formic acid ethyl ester (20 , 81 g, 280.99 mmol) in THF (100 ml) was added over 45 minutes. After 2 hours at 50-55 ^° C., a second portion of NaH (1.35 g) and HCO ₂ Et (10.4 g) are added, followed by a third portion of HCO ₂ Et after 30 minutes (unreacted nitrile is inert in the next step and can be regenerated in the first purification step). The thick paste is stirred overnight and allowed to cool to room temperature. The volatile material was evaporated under reduced pressure, the remaining pale yellow crust was dissolved in a minimum volume of cold water (about 150 ml) at 0 ^° C. The solution was adjusted to pH 6.0 by adding 6N HCl and extracted with CHCl ₃ (3 x 100 ml) . The extract was washed with water, dried over Na ₂ SO ₄ and evaporated in vacuo to a thick amber oil. This crude product is used in the next reaction without further purification.

Example 161. Glycine methyl ester hydrochloride (9.35 g, 74.47 mmol) and anhydrous sodium acetate (6.10 g, 74.47 mmol) were added to the crude formyl derivative of Example 160 (8.9 g, 49.65 mmol) ) in a mixture of MeOH / H ₂ O (4: 1.250 ml). After 24 hours, MeOH was evaporated in vacuo, the mixture of water and oil was extracted with CHCl ₃ . The CHCl ₃ layer was dried (Na ₂ SO ₄ ) and evaporated to give an amber oil, which was fed to a silica gel column. Elution with CHCl ₃ gives two main layers: 3-cycloheptyl propionitrile (used as starting material in the previous step); the desired enamine, which is recrystallized from a mixture of CHCl ₃ / Et ₂ O; yield 6.18 g, mp 57-58 ^o C.

Example 162. In a nitrogen atmosphere, chloroformic acid ethyl ester (4.01 g, 37.03 mmol) was added dropwise to the enamine solution of Example 161 (6.18 g, 24.69 mmol) and DBN (9.19 g, 74, 04 mmol) in dry CH ₂ Cl ₂ (100 ml) with external cooling in an ice bath. After stirring at 0 ^° C. for 1 h, the solution was allowed to stand at room temperature overnight. After checking the reaction progress by TCX, additional ClCO ₂ Et (0.5 ml) and DBN (3.0 ml) were added to complete the reaction, the solution was allowed to stand for 24 hours. The volatile material was evaporated in vacuo, the viscous residue was purified on a short column silica gel (the main purpose of which is to remove a less mobile DBN), obtaining N-protected pyrrole, which is used for the next step without further purification.

Example 163. To a solution of the N-protected pyrrole of Example 162 (7.8 g, 24.19 mmol) in MeOH (100 ml) was added solid Na ₂ CO ₃ (6.41 g, 60.48 mmol), the reaction mixture was stirred at room temperature for 48 h with separation of the resulting unlocked pyrrole. The mixture was evaporated to dryness, the residue was thoroughly triturated with water (50 ml) to dissolve inorganic substances and extracted with CHCl ₃ (3 x 100 ml). The extract was dried (Na ₂ SO ₄ ) and evaporated to give a viscous resin, which was purified by column chromatography using silica gel / CHCl ₃ ; yield 4 g, mp 88- 89 ^o C.

Example 164. Benzoylisothiocyanate (1.5 g, 8.96 mmol) was added dropwise to a solution of the unblocked pyrrole of Example 163 (1.99 g, 7.95 mmol) in dry CH ₂ Cl ₂ (50 ml). After 1 h at room temperature, the solution was evaporated and the gummy residue was dissolved in Et ₂ O (100 ml) with an almost immediate separation of the crystalline solid. The mother liquor Et ₂ O is heated to boiling and diluted with an equal volume of hot cyclohexane. Upon slow cooling, the solution gives an additional thioureido compound; yield 2.89 g (88%), mp. 158-159 ^o C.

Example 165. Methyl iodide (1.7 g, 11.96 mmol) was added to a solution of the thioureido compound of Example 164 (1.7 g, 4.1 mmol) and DBN (0.56 g, 4.52 mmol) in dry CH ₂ Cl ₂ (80 ml) at 0 ^° C. The solution was stirred at 0 ^° C. for 15 minutes, at ambient temperature for 1 hour, then evaporated in vacuo. A solution of the residue in CHCl _{3 was} chromatographed on a silica gel column with CHCl ₃ as an elution solvent, to obtain homogeneous fractions of the methylthio compound as an intermediate.

 Example 166

Раствор метилтиосоединения примера 165 (1,72 г, 4,02 ммоль) в 50 мл MeOH, который был насыщен NH₃ при 0^oC, нагревают при 90-95^oC в течение 24 ч в футерованной стеклянными плитками бомбе из нержавеющей стали. Содержимое охлажденной бомбы выпаривают в вакууме, получая смесь вещества (IVE), бензамида и побочного продукта, который является 2-метилтиопроизводным, в отличие от 2-аминосоединения (IVE). Смесь энергично перемешивают с приблизительно 75 мл Et₂O, нерастворимое белое твердое вещество отфильтровывают и промывают Et₂O. Фильтрат содержал большую часть бензамида и 2-метилтиосоединения. Раствор нерастворимого в Et₂O твердого вещества (0,850 г) в MeOH выпаривают с приблизительно 10 г силикагеля. Измельченный в порошок остаток наносят равномерно на верхнюю часть колонны с силикагелем, которую затем элюируют с помощью смеси CHCl₃/MeOH/HOAc (95:5:1), получая 2-метилтиопроизводное как побочный продукт и желаемое 2-аминосоединение (IVE). (IVE) продукт перекристаллизовывают при экстракции в кипящем изопропилацетате в аппарате Сокслета. Белые кристаллы собирают в три порции и высушивают в вакууме над P₂O₅ при 110^oC в течение 7 ч; выход 54% т.пл. более 300^oC (разл.).

A solution of the methylthio compound of Example 165 (1.72 g, 4.02 mmol) in 50 ml of MeOH that was saturated with NH ₃ at 0 ^° C was heated at 90-95 ^° C for 24 hours in a glass-lined stainless steel bomb. The contents of the cooled bomb are evaporated in vacuo to give a mixture of a substance (IVE), benzamide and a by-product that is 2-methylthio derivative, as opposed to the 2-amino compound (IVE). The mixture was vigorously stirred with approximately 75 ml Et ₂ O, the insoluble white solid was filtered off and washed with Et ₂ O. The filtrate contained most of the benzamide and 2-methylthio compound. A solution of Et ₂ O insoluble solid (0.850 g) in MeOH was evaporated with approximately 10 g of silica gel. The powder residue was uniformly applied to the top of the silica gel column, which was then eluted with a mixture of CHCl ₃ / MeOH / HOAc (95: 5: 1) to give the 2-methylthio derivative as a by-product and the desired 2-amino compound (IVE). (IVE) the product is recrystallized by extraction in boiling isopropyl acetate in a Soxhlet apparatus. The white crystals are collected in three portions and dried in vacuo over P ₂ O ₅ at 110 ^° C. for 7 hours; yield 54% mp more than 300 ^o C (decomp.).

Analysis for C ₁₄ H ₂₀ N ₄ O: Calculated: C 64.60; H 7.74; N, 21.52; Found: C, 64.78; H 8.01; N, 21.61.

Example 167. The substance of Example 166 was tested for enzymatic inhibitory activity as in Example 105. At 1 mM phosphate, IC _{50 was} 0.030 μM, and at 50 mM phosphate, IC _{50 was} 0.840 μM.

 Using the method of Example 97, 3- (1-norbornanyl) propionitrile is obtained from 1-bromnorbornane, and 3- (2-norbornanyl) propionitrile (mixture of 2-ex and 2-endo) is obtained from 2-bromnorbornane. Following examples 98-104, propionitriles are converted into substances (IVF), (IVG) and (IVH).

 Example 169. Using the method of Example 97, 3- (1-bicyclo [3.2.1] octanyl) propionitrile, 3- (2-bicyclo [3.2.1] octanyl) propionitrile, 3- (3-bicyclo [3.2.1 ] octanyl) propionitrile and 3- (8-bicyclo [3.2.1] octanyl) propionitrile are obtained respectively from 1-bromobicyclo [3.2.1] octane, 2-bromobicyclo [3.2.1] octane, 3-bromobicyclo [3.2. 1] octane and 8-bromobicyclo [3.2.1] octane. Following examples 98-104, propionitriles are converted to substance (IVI) and related 2-bicyclo [3.2.1] octanyl-; 3-bicyclo [3.2.1] octanyl and 8-bicyclo [3.2.1] octanyl derivatives.

 Example 170. Using the methodology of the method described in D. Farcasiu, Synthesis, 615 (1972), 6-bicyclo [3.2.1] octane-carboxaldehyde is prepared by reacting bicyclo [3.2.1] octan-6-one with trimethyl sulfoxonium iodide to form an intermediate epoxide, which is then converted to aldehyde by treatment with boron trifluoride etherate. Following the method of Netherlands Pat. 6, 610, 204, the aldehyde was condensed with cyanacetic acid under reflux in a pyridine / toluene solution with a catalytic amount of ammonium acetate for 48-72 hours to obtain the corresponding acrylonitrile. Acrylonitrile is then hydrogenated using palladium-carbon as a catalyst in methanol, as described by Profitt et al. in J. Org. Chem. 40, 127, (1975), to obtain 3- (6-bicyclo [3.2.1] octanyl) propionitrile. Following examples 98-104, propionitrile is converted into a 6-bicyclo [3.2.1] octanyl derivative, similar to the substance (IVL).

 Example 171. Using the method of Example 97, 3- (1-bicyclo [3.3.1] nonanyl) propionitrile and 3- (3-bicyclo [3.3.1] nonanyl) propionitrile are respectively obtained from 1-bromocycle [3.3.1] nonana and 3-bromobicyclo [3.3.1] nonana. Following examples 98-104, propionitriles are converted into a substance (IVM) and a related 3-bicyclo [3.3.1] nonanyl derivative.

 Example 172. Following the method of example 171, bicyclo [3.3.1] nonan-9-one reacts with the formation of the corresponding aldehyde, from which the corresponding 3-substituted propionitrile is obtained, which is then converted to the 9-bicyclo [3.3.1] nonanyl derivative similar to substance (IVM).

 Example 173. Following the method of Example 170, 2-bicyclo [3.3.1] nonanecarboxaldehyde reacts with the formation of the corresponding 3-substituted propionitrile, which is then converted into a 2-bicyclo [3.3.1] nonanil derivative similar to substance (IVM).

 Example 174. Using the method of Example 97, 3- (1-noradamantyl) propionitrile is obtained from 1-bromoradamantane and 3- (2-noradamantyl) propionitrile is obtained from 2-bromoradamantane. Following examples 98-104, propionitrile is converted to the final substance (IVN).

 Example 175. Following the method of Example 170, 3-noradamantanecarboxaldehyde is reacted to produce the corresponding 3-substituted propionitrile, which is then converted to a 3-noradamantyl derivative similar to substance (IVN).

 Example 176. Following the method of Example 170, noradamantan-7-one reacts with the formation of the corresponding aldehyde, from which the corresponding 3-substituted propionitrile is obtained, which is then converted to the 7-noradamantyl derivative similar to substance (IVN).

 Example 177. Using the method of Example 97, 3- (1-bicyclo [2.2.2] octanyl) propionitrile is prepared from 1-bromobicyclo [2.2.2] octane and 3- (2-bicyclo [2.2.2] octanyl) propionitrile obtained from 2-bromobicyclo [2.2.2] octane. Following examples 98-104, propionitriles are converted into a substance (IVK) and a similar 2-bicyclo [2.2.2] octanyl derivative.

 Example 178. Using the method of Example 97, 3- (1-norbornenyl) propionitrile is obtained from 1-bromnorbornene. Following examples 98-104, propionitrile is converted into substance (IVI).

 Example 179. Following the method of Example 170, 5-norbornene-2-carboxaldehyde (a mixture of 2-endo and 2-exo) is reacted to form the corresponding 3-substituted propionitrile, which is then converted to substance (IVJ).

 Example 180

Приведенное выше промежуточное соединение получают в этом примере по методике способа Schiemenz G.P. Engelhard Н. (Chem. Ber. 1962, 95, 195).

The above intermediate compound is prepared in this example by the method of the Schiemenz GP Engelhard N. method (Chem. Ber. 1962, 95, 195).

A mixture of cyanoacetic acid (25.38 g, 298.38 mmol), 2,3,5-trichlorobenzaldehyde (25.0 g, 119.35 mmol), ammonium acetate (500 mg), toluene (120 ml) and pyridine (65 ml) is heated at the boiling temperature under reflux for 16 hours in a flask equipped with a Dean-Stark trap and a refrigerator. The solvents were evaporated in vacuo, the residue was extracted with CHCl ₃ , washed with water, dried (Na ₂ SO ₄ ), and evaporated to give a crude product, which was purified by silica gel column chromatography using hexane-EtOAc as an elution solvent. Yield 23.69 g (73%), mp. 90-91 ^o C.

 Example 181

В этом примере получают упомянутое промежуточное соединение. К перемешиваемой смеси NaH (1,56 г, 65,05 ммоль) и этилового эфира муравьиной кислоты (14,78 г, 199,51 ммоль) в THF (100 мл) добавляют замещенный пентандинитрил примера 180 (10,17 г, 37,17 ммоль) при комнатной температуре в атмосфере азота, полученную реакционную смесь перемешивают в течение 24 ч. Летучее вещество выпаривают в вакууме при комнатной температуре. Воду (50 мл) добавляют к остатку при 0-5^oC и раствор доводят до pH 5-6 20%-ной концентрации HCl (о/о). Тяжелое масло экстрагируют этилацетатом, промывают водой (1 x 100 мл) и высушивают (MgSO₄). Слой этилацетата выпаривают, получая красно-коричневое масло (11,0 г), которое используют в следующей стадии без дальнейшей очистки.

In this example, the aforementioned intermediate is obtained. To a stirred mixture of NaH (1.56 g, 65.05 mmol) and formic acid ethyl ester (14.78 g, 199.51 mmol) in THF (100 ml) was added the substituted pentanedinitrile of Example 180 (10.17 g, 37, 17 mmol) at room temperature in a nitrogen atmosphere, the resulting reaction mixture was stirred for 24 hours. The volatile material was evaporated in vacuo at room temperature. Water (50 ml) was added to the residue at 0-5 ^° C. and the solution was adjusted to pH 5-6 with a 20% concentration of HCl (v / v). The heavy oil was extracted with ethyl acetate, washed with water (1 x 100 ml) and dried (MgSO ₄ ). The ethyl acetate layer was evaporated, yielding a red-brown oil (11.0 g), which was used in the next step without further purification.

 Example 182

В этом примере получают приведенное промежуточное вещество. Гидрохлорид метилового эфира глицина (8,17 г, 65,06 ммоль) и ацетат натрия (5,33 г, 65,06 ммоль) добавляют к раствору сырого формильного соединения примера 181 (11,0 г) в смеси MeOH (80 мл) и воды (20 мл), полученный раствор перемешивают при комнатной температуре в течение 22 ч. После испарения растворителя при комнатной температуре остаток экстрагируют этилацетатом. Промытый (H₂O) и высушенный (MgSO₄) органический слой выпаривают, получая масло. Испарительная колоночная хроматография (силикагель), использующая CHCl₃ в качестве растворителя для элюирования, дает чистый желаемый енамин в виде смеси цис-транс изомеров, которые перекристаллизовывают из MeOH; выход 10,41 г (75%), т.пл. 142-143^oC.

In this example, the recited intermediate is obtained. Glycine methyl ester hydrochloride (8.17 g, 65.06 mmol) and sodium acetate (5.33 g, 65.06 mmol) are added to a solution of the crude formyl compound of Example 181 (11.0 g) in a MeOH mixture (80 ml) and water (20 ml), the resulting solution was stirred at room temperature for 22 hours. After evaporation of the solvent at room temperature, the residue was extracted with ethyl acetate. The washed (H ₂ O) and dried (MgSO ₄ ) organic layer was evaporated to give an oil. Evaporative column chromatography (silica gel) using CHCl ₃ as an elution solvent afforded the pure desired enamine as a mixture of cis-trans isomers which recrystallized from MeOH; yield 10.41 g (75%), mp. 142-143 ^o C.

 Example 183

В этом примере получают приведенное промежуточное вещество. Раствор енамина примера 182 (10,0 г, 26,84 ммоль) в сухом CH₂Cl₂ (100 мл) охлаждают до 0^oC и обрабатывают 1,5-диазабицикло[4.3.0] нон-5-еном (10,53 г, 84,79 ммоль) в атмосфере азота, за которым следует этиловый эфир хлормуравьиной кислоты (6,90 г, 63,57 ммоль). Раствор перемешивают при 0^oC в течение 1 ч, затем при комнатной температуре в течение 48 ч. Летучие вещества испаряют в вакууме, получая вязкую темную смолу, которую очищают испарительной колоночной хроматографией над силикагелем, используя CHCl₃ в качестве растворителя для элюирования. Все фракции, содержащие желаемый N-защищенный пиррол, объединяют и выпаривают, получая пенистое, легкое, бледно-желтое вещество, которое перемешивают в MeOH (100 мл), получая кристаллическое вещество, которое перекристаллизовывают из CHCl₃-MeOH; выход 8,92 г (74,7%), т.пл. 160-161^oC.

In this example, the recited intermediate is obtained. A solution of the enamine of Example 182 (10.0 g, 26.84 mmol) in dry CH ₂ Cl ₂ (100 ml) was cooled to 0 ^° C and treated with 1,5-diazabicyclo [4.3.0] non-5-ene (10, 53 g, 84.79 mmol) in a nitrogen atmosphere, followed by ethyl chloroformate (6.90 g, 63.57 mmol). The solution was stirred at 0 ^° C. for 1 hour, then at room temperature for 48 hours. Volatiles were evaporated in vacuo to give a viscous dark gum which was purified by flash column chromatography over silica gel using CHCl ₃ as an elution solvent. All fractions containing the desired N-protected pyrrole were combined and evaporated to give a foamy, light, pale yellow substance which was stirred in MeOH (100 ml) to give a crystalline substance which was recrystallized from CHCl ₃ -MeOH; yield 8.92 g (74.7%), mp. 160-161 ^o C.

 Example 184

В этом примере получают приведенное промежуточное вещество. Суспензию N-защищенного пиррола примера 183 (8,6 г, 19,34 ммоль) в MeOH (300 мл) обрабатывают Na₂CO₃ (5,12 г, 48,34 ммоль), реакционную смесь перемешивают при комнатной температуре в течение 17 ч с отделением деблокированного пиррола в течение первого часа. Твердый карбонат натрия удаляют фильтрованием и хорошо промывают MeOH. Фильтрат уменьшают до объема 25 мл, выдерживают в холодильнике в течение 1 ч, получая 5,23 г кристаллического продукта. Дальнейшая концентрация маточного раствора дает дополнительные 0,14 г чистого продукта; общий выход 6,45 г (89,5%), т.пл. 178-181^oC.

In this example, the recited intermediate is obtained. A suspension of the N-protected pyrrole of Example 183 (8.6 g, 19.34 mmol) in MeOH (300 ml) was treated with Na ₂ CO ₃ (5.12 g, 48.34 mmol), the reaction mixture was stirred at room temperature for 17 h with separation of the released pyrrole during the first hour. Solid sodium carbonate was removed by filtration and washed well with MeOH. The filtrate was reduced to a volume of 25 ml, kept in the refrigerator for 1 h, obtaining 5.23 g of a crystalline product. Further concentration of the mother liquor gives an additional 0.14 g of pure product; total yield 6.45 g (89.5%), mp. 178-181 ^o C.

 Example 185

В этом примере получают приведенное промежуточное вещество. К суспензии пиррола примера 184 (5,83 г, 15,64 ммоль) в дихлорметане (100 мл) добавляют бензоилизотиоцианат (2,88 г, 17,64 ммоль) при комнатной температуре под азотом. Реакционную смесь перемешивают в течение 30 мин с отделением желаемого тиоуреидосоединения. Добавляют дополнительное количество бензоилизотиоцианата (0,5 мл) и снова перемешивают в течение 30 мин. Растворитель выпаривают досуха и легкий желтый остаток растирают в порошок с метанолом. Белое кристаллическое вещество выделяют фильтрованием и перекристаллизовывают из смеси хлороформ - эфир, получая требуемое тиоуреидопроизводное в виде аналитически чистого образца; выход 7,71 г (92%), т.пл. 210-211^oC.

In this example, the recited intermediate is obtained. To a suspension of the pyrrole of Example 184 (5.83 g, 15.64 mmol) in dichloromethane (100 ml) was added benzoylisothiocyanate (2.88 g, 17.64 mmol) at room temperature under nitrogen. The reaction mixture was stirred for 30 minutes to separate the desired thioureido compound. An additional amount of benzoylisothiocyanate (0.5 ml) was added and stirred again for 30 minutes. The solvent was evaporated to dryness and a light yellow residue was triturated with methanol. A white crystalline substance is isolated by filtration and recrystallized from a mixture of chloroform - ether, obtaining the desired thioureide derivative in the form of an analytically pure sample; yield 7.71 g (92%), mp. 210-211 ^o C.

 Example 186.

В этом примере получают приведенное промежуточное вещество. Раствор тиоуреидопроизводного примера 185 (6,75 г, 12,6 ммоль) и 1,5-диазабицикло[4.3.0] нон-5-ена (1,76 г, 14,20 ммоль) в сухом CH₂Cl₂ (200 мл) охлаждают до 0^oC и обрабатывают йодистым метилом (5,20 г, 36,65 ммоль). Реакционную смесь перемешивают при 0^oC в течение 10 мин и затем в течение 1 ч при комнатной температуре. Растворитель выпаривают при комнатной температуре, остаток экстрагируют CHCl₃, промывают водой (2 x 50 мл), сушат (Na₂SO₄) и выпаривают, получая стекловидную пену (6,95 г), которую используют в следующей стадии без очистки.

In this example, the recited intermediate is obtained. A solution of the thioureide derivative of Example 185 (6.75 g, 12.6 mmol) and 1,5-diazabicyclo [4.3.0] non-5-ene (1.76 g, 14.20 mmol) in dry CH ₂ Cl ₂ (200 ml) is cooled to 0 ^o C and treated with methyl iodide (5.20 g, 36.65 mmol). The reaction mixture was stirred at 0 ^° C. for 10 minutes and then for 1 hour at room temperature. The solvent was evaporated at room temperature, the residue was extracted with CHCl ₃ , washed with water (2 x 50 ml), dried (Na ₂ SO ₄ ) and evaporated to give a glassy foam (6.95 g), which was used in the next step without purification.

 Example 187

В этом примере получают приведенные вещества A и B. Вещество A вещество настоящего изобретения, вещество B промежуточное соединение. Раствор метилтиопроизводного промежуточного вещества примера 186 (6,90 г, 12,54 ммоль) в MeOH (200 мл) насыщают при 0^oC аммиаком и нагревают при 100^oC в течение 20 ч в футерованной стеклянными плитками бомбе из нержавеющей стали. Реакционную смесь доводят до комнатной температуры и растворитель выпаривают при комнатной температуре. Очистка сырой смеси испарительной колоночной хроматографией над силикагелем, используя CHCl₃ в качестве растворителя для элюирования, дает 8B (1,1 г, 21%), т.пл. 290-291^oC, затем CHCl₃-MeOH (95:5) дает чистый 8A (2,76 г, 57,5%), т.пл. 284-285^oC.

In this example, the following substances A and B are prepared. Substance A is a substance of the present invention, substance B is an intermediate. A solution of the methylthio derivative of Example 186 (6.90 g, 12.54 mmol) in MeOH (200 ml) was saturated at 0 ^° C. with ammonia and heated at 100 ^° C. for 20 hours in a glass-lined stainless steel bomb. The reaction mixture was brought to room temperature and the solvent was evaporated at room temperature. Purification of the crude mixture by flash column chromatography over silica gel using CHCl ₃ as an elution solvent afforded 8B (1.1 g, 21%), mp. 290-291 ^o C, then CHCl ₃ -MeOH (95: 5) gives pure 8A (2.76 g, 57.5%), so pl. 284-285 ^o C.

MS (EI) 381 (M ⁺ ), 346 (381-Cl) ⁺ , 341 (381-CH ₂ CN) ⁺ ; IR (KBr) 3323, 3180, 3162 2255, 1685, 1630, 1604, 1558, 1525, 1414, 1393; ¹ H-NMR (DMSO-d ₆ ) δ 3.27-3.50 (m, 2H, CH ₂ CN), 4.86 (t, 1H, CH-CH ₂ CN), 5.90 (s, 2H , NH ₂ ), 7.15 (d, J = 2 Hz, 1H, pyrrole ring CH), 7.57 (d, J = 1.5 Hz, aromatic CH), 7.79 (d, J = 1 7 Hz, 1H, aromatic CH), 10.50 (s, 1H, NH-CO), 11..58 (s, 1H, NH pyrrole ring).

Analysis: calculated for C ₁₅ H ₁₀ Cl ₃ N ₅ O: C, 47.08; H 2.63; N, 18.30; Found: C, 46.92; H 2.94; N, 17.05.

Example 188. The invention of Example 187 was tested for enzymatic inhibitory activity. An enzymatic assay of purinucleoside phosphorylase (PNP) is carried out, in which the PNP activity (IC ₅₀ ) for substance (8A) is determined by radiochemical measurement of the formation of [ ¹⁴ C] -poxanthine from [ ¹⁴ C] -inosine (Biomedicine, 1980, 33, 39) using calf spleen as a source of enzyme. At 1 mM phosphate, the IC ₅₀ is 0.64 μM; at 50 mM phosphate, the IC ₅₀ is 10 μM.

 Example 189

Следуя способу, приведенному в примерах 180-187, 3-(3-хлорфенил)-3-(2-амино-4-оксо-3H, 5H-пирроло[3,2-d] пиримидин-7-ил)пропаннитрил получают, используя 3-(3-хлорфенил)-пентандинитрил в качестве исходного вещества; выход 54,5% т.пл. 157-158^oC.

Following the method described in examples 180-187, 3- (3-chlorophenyl) -3- (2-amino-4-oxo-3H, 5H-pyrrolo [3,2-d] pyrimidin-7-yl) propannitrile was obtained, using 3- (3-chlorophenyl) pentanedinitrile as the starting material; yield 54.5% mp 157-158 ^o C.

MS (FAB), m / z 314 (M + H) ⁺ , 273 (313-CH ₂ CN) ⁺ ; IR (KBr) 3313, 3166, 3148, 2920, 2240, 1682, 1625, 1596, 1573, 1525; ¹ H-NMR (DMSO-d ₆ ) δ 3.24-3.48 (m, 2H, CH ₂ CN), 4.41 (t, 1H, CH-CH ₂ CN), 5.88 (s, 2H , NH ₂ ), 7.13 (d, J = 3.5 Hz, 1H, pyrrole ring CH), 7.20-7.50 (m, 4H, aromatic CH), 10.44 (s, 1H, NHCO), 11.45 (d, 1H, NH pyrrole ring).

Analysis: calculated for C ₁₅ H ₁₂ ClN ₅ O • 0.2MeOH: C 57.02; H 4.03; N, 21.87; Found: C, 57.03; H 3.95; N, 21.88.

3-Aрил-3-(2-амино-4-оксо-3H,5H-пирроло[3,2-d]-пиримидин-7-ил)пропаннитрил
Где Ar имеет одно из следующих значений: (1) фенил, 2,3-дихлорфенил, 3-метилфенил и 3-метоксифенил; (2) тиенил (2- и 3-); (3) фуранил (2- и 3-); (4) пиридинил (2-, 3- и 4-); (5) пирролил (2- и 3-); (6) тиазолил (2-, 4- и 5-); (7) 2-пиразинил; (8) пиридазинил (3- и 4-); (9) пиразолил.Example 190. Following the method described in examples 180-187, the following groups of substances (1-9) of the general formula can be obtained:

3-Aryl-3- (2-amino-4-oxo-3H, 5H-pyrrolo [3,2-d] pyrimidin-7-yl) propannitrile
Where Ar has one of the following meanings: (1) phenyl, 2,3-dichlorophenyl, 3-methylphenyl and 3-methoxyphenyl; (2) thienyl (2- and 3-); (3) furanyl (2- and 3-); (4) pyridinyl (2-, 3- and 4-); (5) pyrrolyl (2- and 3-); (6) thiazolyl (2-, 4- and 5-); (7) 2-pyrazinyl; (8) pyridazinyl (3- and 4-); (9) pyrazolyl.

Example 191. Following the method described in examples 180-187, the following materials 10-14 and 21 can be prepared based on the correspondingly substituted pentanedinitrile. Substances 15-20 and 22 are obtained from the corresponding unsaturated Ar analogues in Example 190. In this method, the nitrile group in the unsaturated analog is converted to the amide group by hydrolysis catalyzed by acid or base, then the unsaturated Ar group is transferred to the saturated R ₂ group by catalytic hydrogenation, followed by re-conversion of the amide to nitrile using known dehydration methods.

3-(Замещенный)-3-(2-амино-4-оксо-3H, 5H-пирроло[3,2-d] -пиримидин-7-ил)пропаннитрил
где R₂ означает: (10) 1-адамантил; (11) 2-адамантил; (12) циклогексил; (13) циклогептил; (14) циклопентил; (15) тетрагидрофуранил; (16) тетрагидротиенил; (17) тетрагидропиранил; (18) пиразолидинил; (19) тиазолидинил; (20) пиперазинил; (21) морфолинил; (22) гексагидропиридазинил.

3- (Substituted) -3- (2-amino-4-oxo-3H, 5H-pyrrolo [3,2-d] pyrimidin-7-yl) propannitrile
where R ₂ means: (10) 1-adamantyl; (11) 2-adamantyl; (12) cyclohexyl; (13) cycloheptyl; (14) cyclopentyl; (15) tetrahydrofuranyl; (16) tetrahydrothienyl; (17) tetrahydropyranyl; (18) pyrazolidinyl; (19) thiazolidinyl; (20) piperazinyl; (21) morpholinyl; (22) hexahydropyridazinyl.

 Example 192.

В этом примере получают приведенное вещество 3-(2-амино-4-оксо-3H,5H-пирроло[3,2-d] пиримидин-7-ил)-3-фенилпропаннитрил. Раствор вещества A, полученного в примере 187 (2,0 г, 5,22 ммоль), в горячем этаноле (250 мл) и диметилформамиде (DMF) (150 мл) гидрируют над 30%-ным Pd/C катализатором (1,0 г) в присутствии триэтиламина (2,64 г, 5,0 эквивалентов) при атмосферном давлении. Спустя 5 ч реакция завершается и катализатор отфильтровывают под давлением N₂. Твердое вещество, полученное после испарения фильтрата, растирают в порошок, разрушают ультразвуком с H₂O и высушивают; выход 1,28 г (88%), т.пл. 168-170^oC.

In this example, the reduced material 3- (2-amino-4-oxo-3H, 5H-pyrrolo [3,2-d] pyrimidin-7-yl) -3-phenylpropanenitrile is obtained. A solution of substance A obtained in Example 187 (2.0 g, 5.22 mmol) in hot ethanol (250 ml) and dimethylformamide (DMF) (150 ml) was hydrogenated over a 30% Pd / C catalyst (1.0 g) in the presence of triethylamine (2.64 g, 5.0 equivalents) at atmospheric pressure. After 5 hours, the reaction is complete and the catalyst is filtered off under N ₂ pressure. The solid obtained after evaporation of the filtrate, triturated, destroyed by ultrasound with H ₂ O and dried; yield 1.28 g (88%), mp. 168-170 ^o C.

MS (FAB), m / z280 (M + H) ⁺ , 239 (280-CH ₂ CN) ⁺ ; ¹ H-NMR (DMSO-d ₆ ) δ 3.20-3.46 (m, 2H, CH ₂ CN), 4.37 (t, 1H, CHCH ₂ CN), 5.86 (s, 2H, NH ₂ ), 7.05 (d, J = 1.5 Hz, 1H, pyrrole ring CH), 7.21 (t, 1H, aromatic CH), 7.30 (t, 2H, aromatic CH), 7 39 (d, J = 7.5 Hz, 2H, aromatic CH), 10.41 (s, 1H, NHCO), 11.40 (s, 1H, pyrrole NH).

Analysis: calculated for C ₁₅ H ₁₃ N ₅ O: C 64.50; H 4.69; N, 25.07; Found: C, 64.68; H 4.57; N, 25.19.

Example 193. The substance obtained in example 192 was tested for enzymatic inhibitory activity as in example 188. At 1 mM phosphate, IC _{50 was} 0.023 μM, and at 50 mM phosphate, IC _{50 was} 4.7 μM.

 Example 194

В этом примере получают приведенное вещество 3-(2-амино-4-оксо-3H,5H-пирроло[3,2-d] пиримидин-7-ил)-3- фенилпропановую кислоту. Раствор вещества, полученного в примере 192 (0,200 г, 0,72 ммоль) в 6N HCl (3,0 мл) нагревают при температуре кипения с обратным холодильником в течение 18 ч. Растворитель выпаривают в вакууме, остаток растирают в порошок с H₂O (6 мл), доводят до pH 10 концентрации гидроксидом аммония. Нерастворимое вещество собирают фильтрованием, фильтрат повторно доводят до pH 6,8. Белое вещество, которое осаждается, собирают, промывают водой и высушивают; выход 0,19 г (89%), т. пл. 290^oC (разл.).

In this example, the reduced material 3- (2-amino-4-oxo-3H, 5H-pyrrolo [3,2-d] pyrimidin-7-yl) -3-phenylpropanoic acid is obtained. A solution of the substance obtained in example 192 (0.200 g, 0.72 mmol) in 6N HCl (3.0 ml) was heated at reflux for 18 hours. The solvent was evaporated in vacuo, the residue was triturated with H ₂ O (6 ml), adjusted to pH 10 with ammonium hydroxide concentration. The insoluble matter was collected by filtration, the filtrate was re-adjusted to a pH of 6.8. The white matter that precipitates is collected, washed with water and dried; yield 0.19 g (89%), mp 290 ^o C (decomp.).

MS (FAB), m / z 299 (M + H) ⁺ ; IR (KBr) 3187, 3181, 1688, 1636, 1557, 1524, 1495, 1398, 1376; UV 0.1 N HCl 234 (17.15), 273 (15.05); pH 7,231 (20.05); 272 (10.72); 0.1 N NaOH 227 (22.3), 264 (7.29), 288 (6.66); ¹ H-NMR (DMSO-d ₆ ) δ 2.86-3.0 (dd, 1H, -CHH-), 3.02-3.14 (dd, 1H, -CHH-), 4.47 (t , 1H, CH-CH ₂ CN), 5.85 (s, 2H, NH ₂ ), 6.95 (d, J = 3 Hz, 1H, pyrrole ring CH), 7.14 (t, 1H, aromatic. CH), 7.24 (t, 2H, aromatic CH), 7.33 (d, J 7.0 Hz, 2H, aromatic CH), 10.38 (shoulder, 1H, NHCO), 11.25 ( shoulder, 1H, NH pyrrole ring), 11.1-12.1 (very broad, visible in the integral, 1H, COOH).

Analysis: calculated for C ₁₅ H ₁₄ N ₄ O ₃ • 0.3H ₂ O: C 59.38; H 4.85; N, 18.45; Found: C, 59.42; H 5.04; N, 18.56.

Example 195. The substance obtained in example 194 was tested for enzymatic inhibitory activity as in example 188. At 1 mM phosphate, IC _{50 was} 0.012 μM, and at 50 mM phosphate, IC _{50 was} 0.19 μM.

 Example 196

В этом примере получают приведенное вещество 3-(2-амино-4-оксо-3H,5H-пирроло[3,2-d] пиримидин-7-ил)-3- фенилпропанамид. Раствор вещества, полученного в примере 192 (0,200 г, 0,72 ммоль) в концентрированной H₂SO₄ (0,5 мл) перемешивают при комнатной температуре в течение 20 ч, затем выливают на дробленый лед (5,0 г) и доводят до pH 6,8 посредством концентрации NH₄OH. Осажденное твердое вещество собирают, промывают водой и высушивают; выход 0,180 г, т.пл. 199-201^oC (разл.).

In this example, the reduced material 3- (2-amino-4-oxo-3H, 5H-pyrrolo [3,2-d] pyrimidin-7-yl) -3-phenylpropanamide is obtained. A solution of the substance obtained in example 192 (0.200 g, 0.72 mmol) in concentrated H ₂ SO ₄ (0.5 ml) was stirred at room temperature for 20 hours, then poured onto crushed ice (5.0 g) and adjusted to a pH of 6.8 by means of a concentration of NH ₄ OH. The precipitated solid is collected, washed with water and dried; yield 0.180 g, mp 199-201 ^o C (decomp.).

MS (FAB), m / z 298 (M + H) ⁺ ; IR (KBr) 3324, 3259, 3185, 3178, 1670, 1626, 1561, 1524, 1411; ¹ H-NMR (DMSO-d ₆ ) δ 2.60 (dd, 1H, -CHH-), 2.86 (dd, 1H, -CHH-), 4.52 (t, 1H, CH-CH ₂ , 5.98 (shoulder, 2H, NH ₂ , 6.67 (s, 2H, CONH ₂ ), 6.99 (d, J 2.5 Hz, 1H, pyrrole ring CH), 7.06-7.31 ( m, 5H, aromatic CH), 10.60 (very broad, 1H, NHCO), 11.37 (shoulder, 1H, NH pyrrole ring).

Analysis: calculated for C ₁₅ H ₁₅ N ₅ O ₂ • 0.8H ₂ O: C 57.80; H 5.36; N, 22.46; Found: C, 57.89; H 5.18; N, 22.36.

Example 197. The substance obtained in example 196 was tested for enzymatic inhibitory activity as in example 188. At 1 mM phosphate, IC _{50 was} 0.20 μM, at 50 mM phosphate, IC _{50 was} 6.6 μM.

 Example 198.

В этом примере получают вышеприведенное вещество 3-(2-амино-4-оксо-3H, 5H-пирроло[3,2-d] пиримидин-7-ил)-3-(3-хлорфенил)пропановую кислоту, метиловый эфир. Хлористый тионил (0,2 г, 0,17 ммоль) добавляют к перемешиваемому метанолу (4,0 мл) при 0^oC. Вещество, полученное в примере 194 (0,2 г, 0,67 ммоль), добавляют, смесь перемешивают при комнатной температуре в течение 18 ч. Растворитель удаляют на водном аспираторе (30^oC) и вакуумном насосе (лиофилизация), получая полутвердую массу, которую очищают на колонне с силикагелем, используя CHCl₃-MeOH в качестве растворителя для элюирования; выход 0,1 г, т.пл. 302-303^oC (разл.)
MS (FAB), m/z 347 (M+H), 273 (346-CH₂CO₂CH₃)⁺; UV 0,1 N HCl 220 (sh), 232 (sh), 273 (15,1), pH 7 буффер 232 (18,1), 270 (7,4), 0,1 N NaOH 262 (6,5), 288 (5,9); ¹H-ЯМР (DMSO-d₆) δ 3,15 (dd, J, 2H, CH₂CO₂CH₃), 3,48 (s, 3H, CH₂CH₃), 4,48 (t, 1H, -CHCH₂-), 5,83 (s, 2, NH₂), 7,06 (d, J, 1H, пиррольное кольцо CH), 7,19, 7,26, 7,39, (комплекс m, 4H, хлорфенил CH), 10,36 (s, 1H, 3-NH), 11,30 (d, J, 1H, 5NH).

In this example, the above material 3- (2-amino-4-oxo-3H, 5H-pyrrolo [3,2-d] pyrimidin-7-yl) -3- (3-chlorophenyl) propanoic acid, methyl ester is obtained. Thionyl chloride (0.2 g, 0.17 mmol) was added to stirred methanol (4.0 ml) at 0 ^° C. The substance obtained in Example 194 (0.2 g, 0.67 mmol) was added, the mixture was stirred. at room temperature for 18 hours. The solvent was removed on an aqueous aspirator (30 ^° C.) and a vacuum pump (lyophilization) to obtain a semi-solid mass, which was purified on a silica gel column using CHCl ₃ -MeOH as an elution solvent; yield 0.1 g, mp 302-303 ^o C (decomp.)
MS (FAB), m / z 347 (M + H), 273 (346-CH ₂ CO ₂ CH ₃ ) ⁺ ; UV 0.1 N HCl 220 (sh), 232 (sh), 273 (15.1), pH 7 buffer 232 (18.1), 270 (7.4), 0.1 N NaOH 262 (6.5 ), 288 (5.9); ¹ H-NMR (DMSO-d ₆ ) δ 3.15 (dd, J, 2H, CH ₂ CO ₂ CH ₃ ), 3.48 (s, 3H, CH ₂ CH ₃ ), 4.48 (t, 1H , -CHCH ₂ -), 5.83 (s, 2, NH ₂ ), 7.06 (d, J, 1H, pyrrole ring CH), 7.19, 7.26, 7.39, (complex m, 4H, chlorophenyl CH), 10.36 (s, 1H, 3-NH), 11.30 (d, J, 1H, 5NH).

Analysis: calculated for C ₁₆ H ₁₅ ClN ₄ O ₃ • 0.3CH ₃ OH: C 54.93; H 4.58; N, 15.72; Found: C, 54.99; H 4.78; N, 15.85.

Example 199. The substance obtained in example 198 was tested for enzymatic inhibitory activity. Significant activity was detected (IC ₅₀ 55 nM).

 Example 200

В этом примере получают 3-(2-амино-4-оксо-3H,5H-пирроло[3,2-d]пиримидин-7-ил)-3-циклогексанпропановую кислоту. Раствор вещества, полученного в примере 194 (83 мг, 0,28 ммоль) в трифторуксусной кислоте (TFA) (15 мл) гидрируют с помощью PtO₂ (83 мг) при давлении 60 фунтов/дюйм² в течение 24 ч. Катализатор отфильтровывают через слой цеолита, фильтрат выпаривают при 25^oC. Остаток суспендируют в воде (8 мл), доводят до pH 8,5 концентрированной NH₄OH и фильтруют через Whatman фильтровальную бумагу для удаления примесей коричневого цвета. Бесцветный фильтрат доводят до pH 6,8, осажденное вещество фильтруют, промывают водой и высушивают; выход 65 мг (77%), т. пл. более 300^oC.

In this example, 3- (2-amino-4-oxo-3H, 5H-pyrrolo [3,2-d] pyrimidin-7-yl) -3-cyclohexane propanoic acid is obtained. A solution of the substance obtained in Example 194 (83 mg, 0.28 mmol) in trifluoroacetic acid (TFA) (15 ml) was hydrogenated with PtO ₂ (83 mg) at 60 lb / ⁱⁿ² for 24 h. The catalyst was filtered off through zeolite layer, the filtrate is evaporated at 25 ^o C. The residue is suspended in water (8 ml), adjusted to pH 8.5 with concentrated NH ₄ OH and filtered through Whatman filter paper to remove brown impurities. The colorless filtrate was adjusted to pH 6.8, the precipitated substance was filtered, washed with water and dried; yield 65 mg (77%), mp more than 300 ^o C.

MS (FAB), m / z 305 (M + H) ⁺ ; IR (KBr) 3330, 3194, 2926, 2852, 1683, 1639, 1557, 1522, 1401, 1377; ¹ H-NMR (DMSO-d ₆ ): 0.85, 1.06, 1.60 (m, 11H, cyclohexyl H), 2.56 (m, 2H, CH ₂ CO ₂ H), 3.05 ( m, 1H, CH-CH ₂ ), 5.79 (s, 2H, NH ₂ ), 6.89 (d, J 2.5 Hz, 1H, pyrrole ring CH), 10.31 (shoulder, 1H, pyrrole NH), 11.16 (s, 1H, NHCO), 11.76 (shoulder, 1H, NH pyrrole).

Analysis: calculated for C ₁₅ H ₂₀ N ₄ O • 1.75H ₂ O: C 53.64; H 7.05; N, 16.68; Found: C, 54.03; H 6.71; N, 16.30.

Example 201. The substance obtained in Example 200 was tested for enzymatic inhibitory activity as in Example 188. At 1 mM phosphate, IC _{50 was} 0.097 μM, and at 50 mM phosphate, IC _{50 was} 1.0 μM.

Example 202. Get the substance of the invention, in which R ₂ -CH ₂ OPO (OH) ₂ . The nitrile group of the substance of Example 192 is converted to the corresponding amide by treatment with sulfuric acid. Using the Hoffmann dehydration reaction, the amide is converted to the corresponding amine, which is then converted to the corresponding pyridinium salt using the pyrrilium salt. The conversion of the salt to the corresponding salt of hydrohalic acid is achieved using sodium bromide, which is then converted to phosphinic acid ester using triethyl phosphite. Hydrolysis of the ester using trimethylsilyl bromide affords the corresponding phosphinic acid.

 Example 203. In this example, the substance of the invention is obtained by changing the number of carbon atoms. The nitrile group of the substance of Example 192 is transferred to the corresponding aldehyde, which is then converted to the corresponding alcohol. Using phosphorus tribromide, the alcohol is converted to the corresponding alkyl bromide, which is then converted to acetonitrile using potassium cyanide.

Example 204. In this example, receive the substance of the invention in which R ₂ CH ₂ OP (O) (OH) ₂ . The alcohol obtained as an intermediate in the previous example is converted to the corresponding diethylphosphonomethyl ether using diethylchloromethylphosphonate. Removal of the ethyl ester group is carried out using trimethylsilyl bromide to give phosphinic acid.

Example 205. In this example, a substance of the invention is obtained in which R ₂ CH ₂ NHSO ₂ NH ₂ . The nitrile group of Example 192 is reduced to an amine using standard catalytic hydrogenation using palladium in an acidic medium (usually 0.01 N to 1N HCl), which is then converted to sulfamide using sulfamoyl chloride.

Example 206. In this example, a substance of the invention is prepared in which R ₂ CH ₂ NHCH ₂ COOH by reaction of the methylamino intermediate obtained in the previous example with chloroacetic acid.

Example 207. In this example, a substance of the invention is prepared in which R ₂ CH ₂ NHPO (OH) ₂ by reacting the intermediate methylamine derivative obtained in Example 206 with diethyl chloromethylphosphonate and treating the resulting product with trimethylsilyl bromide.

Example 208. In this example, a compound of the invention is obtained in which R ₃ OSO ₂ NH ₂ , by reacting the intermediate alcohol obtained in Example 203 with sulfamoyl chloride.

Example 209. In this example, a compound of the invention is obtained in which R ¹ H, R ² phenyl, R ³ CH ₂ CN. Use the methodology of the method described in Mu Ill Lim. et al. J. Org. Chem. vol. 44, N 22, 3826 (1979). The substance of Example 184 and dimethylformamide dimethylacetal are reacted at room temperature for 2 days and then evaporated to dryness in vacuo. The residue crystallizes to give a pure derivative of N- (dimethylamino) methylene, which is cyclized in the presence of a saturated methanolic ammonia solution to give the desired end product.

Example 210. In this example, receive the substance of the invention in which R ¹ OCH ₃ , R ² phenyl, R ³ CH ₂ CN. Using the substance B of Example 187, the S-methyl group is oxidized to methyl sulfone, which is then converted to the final methoxy compound by treatment with sodium methoxide in methanol.

Example 211. In this example, receive a compound of the invention in which R ² tetrazole. The substance of Example 192 was treated with lithium azide in the presence of ammonium chloride as a catalyst in dimethylformamide (DMF) at 100 ^° C. to give the desired tetrazole.

Example 212. In this example, a compound of the invention is obtained in which R ^{2 is} triazole. The substance of example 198 is treated with hydrazine hydrate to give the corresponding hydrazide, which is then treated with an imoester to give the desired triazole.

Example 213. The substance obtained in example 189 was tested for enzymatic inhibitory activity as in example 188. At 1 mM phosphate, IC _{50 was} 0.012 μM, and at 50 mM phosphate, IC _{50 was} 2.0 μM.

Example 214. In this example, receive the substance of the invention in which R ₃ in the above General formula CNH (NH ₂ ). Substance A from Example 187 reacts with sodium methoxide in methanol at room temperature for about 2 days to form the methyl imidate intermediate. The intermediate then reacts with ammonia in methanol to form the corresponding imidine.

Размешанную суспензию метилового сложного эфира примера 198 (6,00 г, 17,3 ммоль) и сухого (NH₄)₂SO₄ (100 мг) в гексаметилдисилазане (400 мл) кипятят с обратным холодильником в течение 8 ч, одновременно обеспечивая защиту от атмосферной влаги. Прозрачный раствор уменьшают в объеме до приблизительно половины его первоначального объема путем дистиллирования при атмосферном давлении, а оставшиеся гексаметилдисалазаны выпаривают в вакууме, получая триметилсилилпроизводное в виде вязкой смолы, которую затем высушивают над P₂O₅. Сырой продукт затем используют в последующей реакции без дальнейшей очистки: MS (FAB), m/z 491 (M+H)⁺.Example 215. 2-amino-7- (3-chlorophenyl-3-hydroxypropyl) -3H, 5H-pyrrolo [3,2-d] pyrimidin-4-one

A mixed suspension of the methyl ester of Example 198 (6.00 g, 17.3 mmol) and dry (NH ₄ ) ₂ SO ₄ (100 mg) in hexamethyldisilazane (400 ml) was refluxed for 8 hours while providing protection against atmospheric moisture. The clear solution was reduced to about half its original volume by distillation at atmospheric pressure, and the remaining hexamethyldisalazanes were evaporated in vacuo to give the trimethylsilyl derivative as a viscous resin, which was then dried over P ₂ O ₅ . The crude product is then used in the subsequent reaction without further purification: MS (FAB), m / z 491 (M + H) ⁺ .

A solution of LiAlH ₄ in tetrahydrofuran (26 ml 1M, 26 mmol) was added dropwise to a solution of trimethylsilylated ester (presumably 17.3 mmol) in dry tetrahydrofuran (200 ml). After stirring for 1 h, the excess hydride was destroyed by the addition of EtOAc (100 ml) and all solvents were evaporated in vacuo. Diluted HCl is added to the stirred suspension of the residue in cold H ₂ O (200 ml) until the pH reaches 1 and after 15 minutes the pH is adjusted to 7 with a NaOH solution. The solid is collected by filtration, washed with cold H ₂ O, slightly dried in vacuo, then powder by stirring with a mixture of 1: 1 (100 ml) cyclohexane and Et ₂ O. The solid is filtered off and partially dried in a funnel under a stream of N ₂ . A solution of the solid in hot MeOH was evaporated to dryness with silica gel (holes 230-400 mesh, 50 g), the mixture was carefully applied to the top of the silica gel column, then eluted with a mixture of CHCl ₃ / MeOH in a ratio of 17: 3 and 4: 1, obtaining desired alcohol (4.65 g, 84%). Using two recrystallizations from H ₂ O / EtOH in a 2: 1 ratio, white crystals are obtained, which are then dried in vacuo over P ₂ O ₅ for 30 hours at 110 ^° C; yield 3.36 g (61%).

MS (FAB), m / z319 (M + H) ⁺ ; 321 (Cl isotopic peak 319) ⁺ , 301 (318-OH) ⁺ , 151 (B + 2H) ⁺ ; UV 0.1 NHCl 233 (17.9), 274 (15.7), pH 7 buffer 230 (22.3), 272 (10.6), 0.1 N NaOH 266 (7.3), 288 ( 6.7); ¹ H-NMR (DMSO-d ₆ ) δ 2.07, 2.21 (2 sextets 1H each, -CHCH ₂ CH ₂ OH), 3.29 (m, 1H, -CH ₂ OH), 4.17 ( t, 1H, -CHCH ₂ CH ₂ OH), 4.45 (t, 1H, -CH ₂ OH), 5.79 (s, 2H, NH ₂ ), 7.09 (d, J, 1H, pyrrole ring CH), 7.18, 7.27, 7.34 (complex m, 4H, chlorophenyl CH), 10.36 (s, 1H, 3-NH), 11.29 (d, J, 1H, 5-NH )

Analysis: calculated for C ₁₅ H ₁₅ ClN ₄ O ₂ : C 56.52; H 4.74; N, 17.58; Found: C, 56.51; H 4.84; N, 17.49.

Раствор спирта из примера 215 (100 мг, 0,314 ммоль) в CF₃COOH (20 мл) гидрогенизируют путем размешивания с платиновым катализатором (100 мг PtO₂) в аппарате Парра при первоначальном давлении в 60 фунтов/дюйм. Катализатор удаляют путем фильтрации через тонкий слой цеолита под N₂. Фильтрат выпаривают, остаток дважды растворяют в MeOH (5,0 мл), а раствор выпаривают. Мутному раствору остатка в H₂O/конц. NH₄OH (20 мл, 1:1) дают постоять в течение 4 ч при комнатной температуре, прежде чем подвергнуть его фильтрованию и выпариванию досуха. Раствор остатка в H₂O (30 мл), содержащий 2 капли 1N NaOH нейтрализуют 1N HCl и охлаждают до 5^oC. Осадившееся белое твердое вещество собирают фильтрацией, промывают холодной водой и высушивают в вакууме над P₂O₅ при температуре 110^oC в течение 6 ч; выход 63 мг (67%), т.пл. 181-186^oC.Example 216. 2-amino-7- (1-cyclohexyl-3-hydroxypropyl) -3H, 5H-pyrrolo [3,2-d] pyrimidin-4-one

A solution of the alcohol from Example 215 (100 mg, 0.314 mmol) in CF ₃ COOH (20 ml) was hydrogenated by stirring with a platinum catalyst (100 mg PtO ₂ ) in a Parr apparatus at an initial pressure of 60 psi. The catalyst is removed by filtration through a thin layer of zeolite under N ₂ . The filtrate was evaporated, the residue was dissolved twice in MeOH (5.0 ml), and the solution was evaporated. A turbid solution of the residue in H ₂ O / conc. NH ₄ OH (20 ml, 1: 1) was allowed to stand at room temperature for 4 hours before being filtered and evaporated to dryness. A solution of the residue in H ₂ O (30 ml) containing 2 drops of 1N NaOH was neutralized with 1N HCl and cooled to 5 ^o C. The precipitated white solid was collected by filtration, washed with cold water and dried in vacuum over P ₂ O ₅ at 110 ^o C within 6 hours; yield 63 mg (67%), mp. 181-186 ^o C.

MS (FAB), m / z 291 (M + H) ⁺ ; UV 0.1 N HCl 236 (16.2), 275 (14.9), pH 7 buffer 231 (18.8), 274 (12.1), 0.1 N NaOH 230 (19.6), 268 (7.8), 288 (shoulder); ¹ H-NMR (DMSO-d ₆ ) δ 0.85, 1.08, 1.59 (complex multiplets, 11H, cyclohexyl CH), 1.78 (m, 2H, -CHCH ₂ CH ₂ OH), 2, 62 (m, 1H, -CHCH ₂ CH ₂ OH), 3.14, 3.17, 3.24 (complex multiplets, 2H, -CH ₂ OH), 4.25 (br. 1H, -CH ₂ OH) 5.82 (s, 2H, -NH ₂ , 6.86 (d, J, 1H, H-6), 10.35 (br 1H, 3-NH), 11.17 (d, J, 1H 5-NH).

Analysis: calculated for C ₁₅ H ₂₂ N ₄ O ₂ • 0.5H ₂ O: C 60.18; H 7.74; N, 18.71; Found: C, 60.09; H 7.68; N, 18.73.

2-Амино-7-[2-циано-1-(3-хлорфенил)этил] -3H, 5H-пирроло[3,2-d] пиримидин-4-он (20 г, 63,75 ммоль) гидролизуют до получения соответствующей кислоты с помощью способа, описанного в примере 194. Выход: 18 г (84,9%), т.пл. 295-296^oC.Example 217. 2-amino-7- [2-carboxy-1- (3-chlorophenyl) ethyl] -3H, 5H-pyrrolo [3,2-d] pyrimidin-4-one

2-amino-7- [2-cyano-1- (3-chlorophenyl) ethyl] -3H, 5H-pyrrolo [3,2-d] pyrimidin-4-one (20 g, 63.75 mmol) is hydrolyzed to obtain the corresponding acid using the method described in example 194. Yield: 18 g (84.9%), so pl. 295-296 ^o C.

MS (FAB), m / z 333 (M + H) ⁺ ; IR (KBr) 3182, 3170, 1686, 1639, 1596, 1572, 1525, 1477, 1402, 1377; UV 0.1 N HCl 235 (17.2), 273 (15.42); pH 7,233 (20.2), 272 (1058), 0.1 N NaOH 231 (20.9), 263 (7.2), 288 (6.67); ¹ H-NMR (DMSO-d ₆ ) δ 2.96 (m, 2H, CH ₂ CN), 4.48 (t, 1H, CH-CH ₂ CN), 6.06 (br 2H, NH ₂ ) 7.11 (d, J 2.6 Hz, 1H, pyrrole ring CH), 7.21 (d, J 8 Hz, 1H, aromatic CH), 7.25-7.46 (m, 3H, aromatic CH), 10.3-11.0 (very broad, visible in the integral, 1H, NH or COOH), 10.42 (br. 1H, NH pyrrole ring), 11.7-12.3 (very wide, visible in the integral, 1H, NH or COOH).

Analysis: calculated for C ₁₅ H ₁₄ N ₄ O ₃ • 0.3H ₂ O: C 59.38; H 4.85; Found: C, 59.42; H 5.04; N, 18.56.

К раствору 3-(2-метилтио-4-оксо-3H,5H-пирроло[3,2-d]пиримидин-7-ил)-3-(3-хлорфенил)пропаннитрила (1,0 г) в этаноле (100 мл) добавляют 30% Pd/C и образовавшуюся суспензию кипятят с обратным холодильником в течение нескольких минут. Затем при постоянном помешивании добавляют гидрат гидразина (300 мл), реакционную смесь кипятят с обратным холодильником в течение 2 дней. Затем добавляют дополнительные порции гидрата гидразина (300 мл) и Pd/C (0,5 г) и реакционную смесь кипятят с обратным холодильником еще 4 дня. Катализатор удаляют фильтрацией, фильтрат уменьшают в объеме до 75 мл, фильтруют и выпаривают, получая желаемый продукт в виде белого твердого вещества; выход 0,82 г (94,7%); т.пл. 230^oC.Example 218. 7- [2-cyano-1- (3-chlorophenyl) ethyl] -3H, 5H-pyrrolo [3,2-d] pyrimidin-4-one

To a solution of 3- (2-methylthio-4-oxo-3H, 5H-pyrrolo [3,2-d] pyrimidin-7-yl) -3- (3-chlorophenyl) propannitrile (1.0 g) in ethanol (100 ml) add 30% Pd / C and the resulting suspension is refluxed for several minutes. Then, with constant stirring, hydrazine hydrate (300 ml) was added, the reaction mixture was refluxed for 2 days. Then, additional portions of hydrazine hydrate (300 ml) and Pd / C (0.5 g) were added and the reaction mixture was refluxed for another 4 days. The catalyst was removed by filtration, the filtrate was reduced to 75 ml, filtered and evaporated to give the desired product as a white solid; yield 0.82 g (94.7%); so pl. 230 ^o C.

MS (FAB), 299 (M + 1) ⁺ ; IR (KBr) 3166, 3155, 3150, 3142, 3133, 3093, 2248, 1673, 1595, 1414; UV (0.1 N HCl), 237 (27.8); pH 7; 262 (9.9); 233 (31, l); 0.1 N NaOH, 268 (8.9); ¹ H-NMR (DMSO-d ₆ ) δ 3.3-3.52 (m, CH ₂ CN), 4.6 (t, 1H, CHCH ₂ -CN), 7.24-7.56 (m, aromatic CH and H-6), 7.84 (s, H-2), 11.95 (s, NHCO), 12.11 (s, pyrrole NH).

Analysis: calculated for C ₁₅ H ₁₁ ClN ₄ O: C, 60.31; H 3.71; N, 18.75; Found: C, 60.19; H 3.92; N, 18.41.

IC ₅₀ for PNP 10 nM.

The table shows the data on the inhibition of proliferation of T-lymphocytes ^a .

^a CCRF-CEM leukemic T lymphocytes.

^b The concentration required to inhibit the proliferation of 50% of the cells compared with untreated control cells.

 The above compounds were not able to inhibit the proliferation of B-lymphocytes at a concentration of 100 μM, thereby demonstrating their inherent selectivity for T-lymphocytes.

Claims (10)

1. Derivatives of 2-amino-7- (CHR ₂ R ₃ ) -3H, 5H-pyrrolo [3,2-d] pyrimidin-4-one of the formula

where R ₁ H or NH ₂ ;
R _{2 is a} saturated five-membered heterocycle containing sulfur or oxygen as a heteroatom, or a six-membered heterocycle containing N as a heteroatom, and which may optionally be substituted with C ₁ C ₄ alkoxy, trifluoromethyl or halogen, C ₆ - C _{1 0} cycloalkyl, unsubstituted or halogenated phenyl;
R ₃ H, (CH ₂ ) _n CN, (CH ₂ ) _n CO ₂ H, (CH ₂ ) _n CONH ₂ , CO ₂ CH ₃ , (CH ₂ ) _n CO ₂ CH ₃ or (CH ₂ ) _n OH, where n 1-4, or R ₂ and R ₃ together with the CH group to which they are attached form a cyclohexyl or cyclohexenyl group. 2. The compound of claim 1, wherein the CHR ₂ R ₃ together form a cyclohexyl or cyclohexenyl group. 3. The compound according to claim 1, where R ₁ NH ₂ . 4. A compound according to Claim. 1, where R ₂ is phenyl. 5. The compound according to claim 1, where R ₃ CH ₂ CN, CH ₂ COOH or CH ₂ CONH ₂ . 6. The compound of claim 1, wherein R _{2 is} optionally substituted with C ₁ -C ₄ alkoxy, halogen or trifluoromethyl 3-piperidinyl, 2-tetrahydrofuranyl, 2-tetrahydrothienyl, 3-, 4-pyridinyl, cyclopentyl, cyclohexyl, cycloheptyl, 2 -adamantyl, phenyl, 3-chlorophenyl, 2,3,5-trichlorophenyl.  7. A method for selectively inhibiting the proliferation of mammalian T-lymphocytes and not affecting B-lymphocytes, characterized in that the mammal is administered compounds according to any one of paragraphs. 1 6 in an effective amount. 8. A method of obtaining derivatives of 2-amino-7- (CHR ₂ R ₃ ) -3H, 5H-pyrrolo [3,2-d] pyrimidin-4-one of the formula

where R ₁ NH ₂ ;
R ₂ phenyl unsubstituted or substituted by halogen or cyclohexyl;
R ₃ CH ₂ CN,
characterized in that the following sequential steps are carried out: a) a reaction of the corresponding R ₂ -substituted cyclic aldehyde with cyanacetic acid in the presence of ammonium acetate to obtain 3 (R ₂ ) -substituted pentanedinitrile, b) a reaction of the obtained 3 (R ₂ ) -substituted pentanedinitrile with formic acid alkyl ester in the presence of scavenging to obtain 3 (R ₂ ) -2-formylpentanedinitrile, c) the latter is reacted with glycine methyl ester hydrochloride in the presence of sodium or ammonium acetate to give methyl N - [(3 (R ₂ ) -2, 4 - dicyano) -2-butenyl] -glycine, d) the methyl N - [(3 (R ₂ ) -2,4-dicyano) -2-butenyl] -glycine obtained in this way is reacted with chloroacetic acid alkyl ester in the presence of 1.5 diazabicyclo [4.3.0] non-5-ene or 1,8-diazabicyclo [5.4.0] undec-7-ene to give methyl 3-amino-4- (2-cyano-1 (R ₂ ) ethyl ) - 1-ethyl-1H-pyrrole-1,2-dicarboxylate, d) the obtained methyl-3-amino-4- (2-cyano-1 (R ₂ ) ethyl) -1-ethyl-1H-pyrrole-1, 2-dicarboxylate is treated with a base to obtain methyl 3-amino-4- (2-cyano-1 (R ₂ ) ethyl) -1H-pyrrolo-2-carboxylate, e) the latter is reacted with benzoylisothiocyanate with by irradiating N-benzoyl-N '- [4- (2-cyano-1 (R ₂ ) ethyl) -2-methoxycarbonyl-1H-pyrrol-3-yl] thiourea; g) the resulting N-benzoyl-N' - [ 4- (2-cyano-1 (R ₂ ) -ethyl) -2-methoxycarbonyl -1H-pyrrol-3-yl] thiourea is alkylated with alkyl halide to give N-benzoyl-N '- [4- (2-cyano-1 (R ₂ ) ethyl) -2-methoxycarbonyl-1H-pyrrol-3-yl] -S-methylthiourea, h) then N-benzoyl-N '- [4- (2-cyano-1 (R ₂ ) -ethyl ) -2-methoxycarbonyl-1H-pyrrol-3-yl] -S-methylthiourea is treated with a methanol or ethanol solution of ammonia to give a mixture of 3 (R ₂ ) -3- [2-amino-4-oxo-3H, 5H-pyrrolo [ 3,2-d] pyrimidin-7-yl] propannitrile and 3 (R ₂ ) -3 - [2-methi lmercapto-4-oxo-3H, 5H-pyrololo [3,2-d] pyrimidin-7-yl] propannitrile. 9. The method according to p. 8, where R ₂ phenyl. 10. A method of obtaining derivatives of 7- (CHR ₂ R ₃ ) -3H, 5H-pyrrolo [3,2-d] pyrimidin-4-one of the formula I

where R ₁ H;
R ₂ phenyl unsubstituted or substituted by halogen, or cyclohexyl;
R ₃ CH ₂ CN,
characterized in that the following sequential steps are carried out: a) a reaction of the corresponding cyclic aldehyde with cyanoacetic acid in the presence of ammonium acetate to obtain 3 (R ₂ ) -substituted pentanedinitrile, b) a reaction of the obtained 3 (R ₂ ) pentanedinitrile with formic acid alkyl ester in in the presence of a base to give 3 (R ₂ ) - 2-formylpentanedinitrile, c) the latter is reacted with glycine methyl ester hydrochloride in the presence of sodium or ammonium acetate to give methyl N - [(3 (R ₂ ) -2,4-dicyano) -2-butenes ] Glycine g) thus obtained methyl -N - [(3 (R ₂₎ 2,4-dicyano) -2-butenyl] glycine is reacted with a chloroacetic acid alkyl ester in the presence of 1,5-diazabicyclo [4.3.0] non-5-ena or 1,8-diazabicyclo [5.4.0] undec-7-ene to give methyl 3-amino-4- (2-cyano-1 (R ₂ ) ethyl) -1-ethyl-1H -pyrrole-1,2-dicarboxylate, d) methyl-3-amino-4- (2-cyano-1 (R ₂ ) -ethyl) -1-ethyl-1H-pyrrole -1,2-dicarboxylate is treated with a base to obtain methyl 3-amino-4- (2-cyano-1 (R ₂ ) -ethyl) -1H-pyrrole-2-carboxylate, e) then methyl-3-amino-4- (2-cyclo-1 (R ₂ ) ethyl) -1H-pyrrolo-2-carboxylate is subjected to vzamodeist vii with dimethylformamide dimethylacetal to give methyl 4- (2-cyano-1 (R ₂ ) ethyl) -3- [N- (dimethylaminomethylene) amino] - 1H-pyrrolo-2-carboxylate, and g) methyl-4- (2-Cyano-1 (R ₂ ) ethyl) -3- [N- (dimethylaminomethylene) amino] -1H-pyrrolo-2-carboxylate is treated with a methanol solution of ammonia to give 3 (R ₂ ) 3- [4-oxo-3H , 5H-pyrrolo [3,2-d] pyrimidin-7-yl] propannitrile. Priority by signs:
10/31/89 N 429,097 compounds according to claim 1, where R ₂ optionally substituted heteroalicyclic group, N 429,098 compounds according to claim 1, where R ₂ optionally substituted cyclohexenyl or cyclohexyl, N 429,100 compounds according to claim 1, where R ₂ optionally substituted pyridinyl , N 429,099 compounds according to claim 1, where R ₂ optionally substituted alicyclic group;
11.29.89 the compound of claim 1, wherein R _{2 is an} optionally substituted cyclic group optionally containing a heteroatom, R ₃ hydrogen, CO ₂ H, CH ₂ CONH ₂ , CO ₂ CH ₃ , CH ₂ CN, CH ₂ CO ₂ H.

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