CA2072123C - Inhibitors of purine nucleoside phosphorylase - Google Patents

Inhibitors of purine nucleoside phosphorylase

Info

Publication number
CA2072123C
CA2072123C CA002072123A CA2072123A CA2072123C CA 2072123 C CA2072123 C CA 2072123C CA 002072123 A CA002072123 A CA 002072123A CA 2072123 A CA2072123 A CA 2072123A CA 2072123 C CA2072123 C CA 2072123C
Authority
CA
Canada
Prior art keywords
compound
cyclo
amino
pyrrolo
pyrimidin
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Expired - Fee Related
Application number
CA002072123A
Other languages
French (fr)
Other versions
CA2072123A1 (en
Inventor
John A. Secrist, Iii
Mark D. Erion
John A. Montgomery
Steven E. Ealick
Wayne C. Guida
Shri Niwas
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Biocryst Pharmaceuticals Inc
Original Assignee
Biocryst Pharmaceuticals Inc
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Priority claimed from US07/429,098 external-priority patent/US4985434A/en
Priority claimed from US07/429,097 external-priority patent/US5008270A/en
Priority claimed from US07/429,100 external-priority patent/US4985433A/en
Priority claimed from US07/429,099 external-priority patent/US5008265A/en
Application filed by Biocryst Pharmaceuticals Inc filed Critical Biocryst Pharmaceuticals Inc
Publication of CA2072123A1 publication Critical patent/CA2072123A1/en
Application granted granted Critical
Publication of CA2072123C publication Critical patent/CA2072123C/en
Anticipated expiration legal-status Critical
Expired - Fee Related legal-status Critical Current

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D487/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00
    • C07D487/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00 in which the condensed system contains two hetero rings
    • C07D487/04—Ortho-condensed systems
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12—Antivirals
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C255/00—Carboxylic acid nitriles
    • C07C255/01—Carboxylic acid nitriles having cyano groups bound to acyclic carbon atoms
    • C07C255/31—Carboxylic acid nitriles having cyano groups bound to acyclic carbon atoms having cyano groups bound to acyclic carbon atoms of a carbon skeleton containing rings other than six-membered aromatic rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C255/00—Carboxylic acid nitriles
    • C07C255/01—Carboxylic acid nitriles having cyano groups bound to acyclic carbon atoms
    • C07C255/32—Carboxylic acid nitriles having cyano groups bound to acyclic carbon atoms having cyano groups bound to acyclic carbon atoms of a carbon skeleton containing at least one six-membered aromatic ring
    • C07C255/35—Carboxylic acid nitriles having cyano groups bound to acyclic carbon atoms having cyano groups bound to acyclic carbon atoms of a carbon skeleton containing at least one six-membered aromatic ring the carbon skeleton being further substituted by halogen atoms, or by nitro or nitroso groups
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C255/00—Carboxylic acid nitriles
    • C07C255/01—Carboxylic acid nitriles having cyano groups bound to acyclic carbon atoms
    • C07C255/32—Carboxylic acid nitriles having cyano groups bound to acyclic carbon atoms having cyano groups bound to acyclic carbon atoms of a carbon skeleton containing at least one six-membered aromatic ring
    • C07C255/40—Carboxylic acid nitriles having cyano groups bound to acyclic carbon atoms having cyano groups bound to acyclic carbon atoms of a carbon skeleton containing at least one six-membered aromatic ring the carbon skeleton being further substituted by doubly-bound oxygen atoms
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C255/00—Carboxylic acid nitriles
    • C07C255/01—Carboxylic acid nitriles having cyano groups bound to acyclic carbon atoms
    • C07C255/32—Carboxylic acid nitriles having cyano groups bound to acyclic carbon atoms having cyano groups bound to acyclic carbon atoms of a carbon skeleton containing at least one six-membered aromatic ring
    • C07C255/42—Carboxylic acid nitriles having cyano groups bound to acyclic carbon atoms having cyano groups bound to acyclic carbon atoms of a carbon skeleton containing at least one six-membered aromatic ring the carbon skeleton being further substituted by singly-bound nitrogen atoms, not being further bound to other hetero atoms
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D207/00—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom
    • C07D207/02—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom with only hydrogen or carbon atoms directly attached to the ring nitrogen atom
    • C07D207/30—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having two double bonds between ring members or between ring members and non-ring members
    • C07D207/34—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having two double bonds between ring members or between ring members and non-ring members with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms

Landscapes

  • Organic Chemistry (AREA)
  • Chemical & Material Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • General Chemical & Material Sciences (AREA)
  • Public Health (AREA)
  • Medicinal Chemistry (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Veterinary Medicine (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Virology (AREA)
  • Communicable Diseases (AREA)
  • Oncology (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Nitrogen Condensed Heterocyclic Rings (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)
  • Pyrrole Compounds (AREA)
  • Organic Low-Molecular-Weight Compounds And Preparation Thereof (AREA)

Abstract

Disclosed is a compound containing a 2-amino-7-(R)-3H,5H-pyrrolo[3,2-a]pyrimidin-4-one wherein K is cyclohexeny cyclohexyl, or -CH2-R1, and wherein R1 is an optionally substituted heteroalicyclic, pyridinyl or alicyclic group. Also dis-closed is a compound of formula (I), wherein R1 is H, NH2, or OCH3, R2 is an optionally substituted cyclic group of 5-carbon atoms optionally containing one or more heteroatoms, R3 and R4 are independently H or C1-4 alkyl, m is 0-4, n is 0-6, p is 0-1, X is CN, CSNH2, PO(OH)2, COOH, SO2NH2, NH2, OH CNHNH2, tetrazole, or triazole, COR5 where R5, C1-4 alkyl, CF3, NH2, or OC1-4 alkyl, and Y is O or NH.

Description

,.-...
0. F ~ W J. l=N
INHIBITORS OF PURINE NL1CI~EOSIDE PHOSPHORYLASE
The present invention relates to derivatives of 2-amino-3H,5H_-pyrrolo[3,2-d_]pyrimidin-4-one and to derivatives of 4-oxo-3H_,5H_-pyrrolo[3,2-d]pyrimidine. It also relates to 4-oxo-3H,5H-pyrrolo[3,2-d]pyrimidine derivatives substituted at the 7-position.
Purine nucleoside phosphorylase (PNP) catalyzes the phosphorolysis of purine nucleosides in a reversible reaction. Individuals who are deficient in PNP exhibit impaired T-cell development, resulting in lowered cell-mediated immunity, but normal B-cell development, resulting in normal humoral immunity. Accordingly, specific inhibitors of PNP that selectively inhibit T-cell development without damaging humoral immunity could be potentially effective against disorders in which activated T-cells are pathogenic. ' As a PNP inhibitor, the present invention provides a 2-amino-7-(R)-3H_,5H_-pyrrolo[3,2-d]gyrimidin-4-one wherein R
is optionally substituted cyclohexenyl, cyclohexyl, or -CH2-R1, wherein R1 is an optionally substituted heteroalicyclic, pyridinyl or alicyclic group.
The present invention is also directed to a compound of the formula ' y)P-(CH2)m X
wherein R1 is H, NH2, or OCH3, R2 is an optionally substituted cyclic group of 5-7 carbon atoms optionally t , containing one or more heteroatoms, R3 and R4 are ' independently H or C1-4 alkyl, m is 0-4, n is 0-6, p is 0-l, X is CN, CSNH2, PO(OH)2, COON, S02NH2, NH2, OH, CNHNH2, ' tetrazole, triazole or CORS where R5 is C1-4 alkyl, CF3, NH2, or OCl-4 alkyl, and Y is 0 or NH. The compound of the present invention is useful as a PNP inhibitor. Also 'contemplated according to the present invention is a pharmaceutical composition for the selective suppression of mammalian T-cell immunity comprising an pharmaceutically effective amount of the compound of the present invention and a pharmaceutically acceptable carrier or diluent and a method for the selective suppression of mammalian T-cell immunity without diminished effect on humoral immunity comprising administering to a subject a pharmaceutically , effective amount of the compound of the present invention.
In one aspect of the invention there is provided a compound 2-amino-7-(R)-3~,5F~-pyrrolo[3,2-d]-pyrimidin-4-one (I) wherein R is and R1 is an optionally substituted heteroalicyclic group.
Preferably, the heteroalicyclic group is a 5 or 6 membered saturated ring having oxygen, nitrogen, or sulfur as the heterocyclic atom. More preferably the heteroalicyclic group is 2- or 3-tetrahydrothienyl, 2-, 3-, or 4-piperidinyl, 2- or 3-tetrahydrofuranyl, 2-, or 3-pyrrolidinyl, or 2-, 3-, or 4-tetrahydropyranyl. In a preferred aspect, R1 is unsubstituted, e.g., the compound (I) is 2-amino-7-(2-piperidinylmethyl)-3H,5H_-pyrrolo[-3,2-d]pyrimidin-4-one (IB), 2-amino-7-(3-piperidinyl-methyl)-3H,5H-pyrrolo[3,2-d]pyrimidin-4-one (IC), 2-amino-7-(4-piperidinylmethyl)-3_H,5H_-pyrrolo[3,2-d]pyrimidin-4-one (ID), 2-amino-7-(2-tetrahydrofuranylmethyl)-3N,5H_-pyr-rolo[3,2-d]pyrimidin-4-one (IE), 2-amino-7-(3-tetrahy-drofuranylmethyl)-3H,5H_-pyrrolo[3,2-d]pyrimidin-4-one (IF), . . ~~'w~~.~~
~1Y ~ ~Tl J. ~~

2-amino-7-(2-tetrahydrothienylmethyl)-31i,5H-pyrrolo[-3,2-d]pyrimidin-4-one (IG), 2-amino-7-(3-tetrahydrothieny-lmethyl)-3H,5H-pyrrolo[3,2-d]pyrimidin-4-one (IH), 2-amino-7-(2-pyrrolidinylmethyl)-3H,5H_-pyrrolo[3,2-d]pyrimidin-4-one (II), 2-amino-7-(3-pyrrolidinylmethyl)-3H,5H-pyr-rolo[3,2-d]pyrimidin-4-one (IJ), 2-amino-7-(2-tetrahydro-pyranylmethyl)-3#~,SIB-pyrrolo[3,2-~,]pyrimidin-4-one (IL), 2-amino-7-(3-tetrahydropyranylmethyl)-3~,5~-pyrrolo[3,2-~]py-rimidin-4-one (IM), or 2-amino-7-(4-tetrahydropyranyl-methyl)-3I~,5H_-pyrrolo[3,2-d_]pyrimidin-4-one (IN). In an alternative preferred embodiment the R1 has one or two substituents selected from the group consisting of halogen, hydroxy, alkoxy, alkyl, or trifluoromethyl. As halogen is preferably mentioned chloro or fluoro. As alkoxy is preferably mentioned lower alkoxy, including methoxy, ethoxy, propoxy and butoxy. As alkyl is preferably mentioned lower alkyl, including methyl, ethyl, propyl and butyl.
In another aspect of the invention there is provided 2o a compound 2-amino-7-(R)-3H_,5H_-pyrrolo[3,2-d]-pyrimidin-4-one (II) wherein R is and R1 is optionally substituted pyridinyl. In a preferred aspect, R1 is unsubstituted, i.e., the compound (II) is 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]-pyrimidin-4-one (IIC). In an alternative preferred embodiment the R1 group has one or two substituents selected from the group consisting of halogen, hydroxy, alkoxy, alkyl, or trifluoromethyl. As halogen is preferably mentioned chloro or fluoro. As alkoxy is preferably mentioned lower alkoxy, including methoxy, ethoxy, propoxy and butoxy. As alkyl is (:~;~
.. '' preferably mentioned lower alkyl, including methyl, ethyl, propyl and butyl.
In another aspect of the invention there is provided a compound (III) 2-amino-7-(R)-3H,5H-pyrrolo[3,2-d]
pyrimidin-4-one wherein the R group is unsubstituted or substituted 1-, 2-, or 3-cyclohexenyl or cyclohexyl. In a preferred aspect, R is unsubstituted, i.e., the compound (III) is 2-amino-7-(1-cyclohexenyl)-3H_,5H_-pyrrolo[3,2-d]
pyrimidin-4-one (IIIA), 2-amino-7-(2-cyclohexenyl)-3I~,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). In an alternative preferred embodiment the R
has at least one substituent selected from the group ~cnsisting of halogen, hydroxy, alkoxy, alkyl, or .=ifluoromethyl. As halogen is preferably mentioned chloro or fluoro. As alkoxy is preferably lower alkoxy, including mathoxy, ethoxy, propoxy and butoxy. As alkyl is preferably mentioned lower alkyl, including methyl, ethyl, propyl and butyl.
In another aspect of the invention there is provided a compound 2-amino-7-(R)-3~,5F~-pyrrolo[3,2-d]-pyrimidin-4-one (IV) wherein R is and R1 is an optionally substituted alicyclic group.
Preferable alicyclic groups include, e.g., single-ring cycloparafins such as cyclopentyl, cyclohexyl, and cycloheptyl, multi-ring cycloparafins such as 1- and 2-adamantyl, 1-norbornanyl, 2-exo-norbornanyl, 2-endo-norbornanyl, 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 1- and 2-norbornenyl. Examples of the preferred compound (IV) are 2-amino-7-(2-adamantylmethyl)-3H,5H-pyrrolo[3,2-d]-., y.
~~ A'~.r9'~
~w.~.d~ r.,~~
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-one (IVC), 2-amino-7-(cyclohexylmethyl)-3H,5H-pyrrolo[3,2-d]-5 pyrimidin-4-one (IVD), 2-amino-7-(cycloheptylmethyl)-3_H,SH-pyrrolo[3,2-d]pyrimidin-4-one (IVE), 2-amino-7-(1-norbornanylmethyl)-3I~,,5~,-pyrrolo[3,2-d]pyrimidin-4-one (IVF), 2-amino-7-(2-exo-norbornanylmethyl)-3H,5~i-pyrrolo[3,2-d]pyrimidin-4-one (IVG), 2-amino-7-(2-endo-norbornanylmethyl)-3H_,5I~,-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]octanyl- methyl)-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-noradamantyl-methyl)-3H_,5H-pyrrolo[3,2-d]-pyrimidin-4-one (IVN). In an alternative preferred embodiment the R1 group has one or two substituents selected from the group consisting of halogen, hydroxy, alkoxy, alkyl, or trifluoromethyl. As halogen is preferably mentioned chloro or fluoro. As alkoxy is preferably mentioned lower alkoxy, including methoxy, ethoxy, propoxy and butoxy. As alkyl is preferably mentioned lower alkyl, including methyl, ethyl, propyl and butyl.
The present invention is also directed to a compound of the formula Ri Y)P (CHZ)m X

t'_:':) ~~?'~ ~~ n~o wherein R1 is H, NH2, or OCH3, R2 is an optionally ' substituted cyclic group optionally containing one or more heteroatoms, R3 and R4 are independently H or C1-4 alkyl, m is 0-4, n is 0-6, p is 0-1, X is CN, CSNH2, PO(OH)2, COOH, S02NH2, NH2, OH, CNHNH2, tetrazole, triazole or CORS where R5 is C1-4 alkyl , CF3 , NH2 , or OCl_4 alkyl , and Y is O or NH.
The optionally substituted cyclic group (hereinafter referred to as cyclo) recited for the above formula l0 includes aromatic, heteroaromatic, alicyclic, and heteroalicyclic groups preferably containing five to nine atoms. Preferred optional substituents include halogen, hydroxy, alkoxy, alkyl, and trifluoromethyl. Exemplary substituents include chloro, fluoro, methoxy, ethoxy,, propoxy, butoxy, methyl, ethyl, propyl, and butyl.
Preferred heteroatoms include oxygen, nitrogen, and sulfur, which can be present in combination in the same group. The preferred aromatic and heteroaromatic groups are phenyl, 2-or 3-thienyl, 2- or 3-furanyl, 2-, 3-, or 4-pyridinyl, 2 or 3-pyrrolyl, 2-, 4-, or 5-thiazolyl, 2-pyrazinyl, 3- or 4-pyridazinyl, and 3-, 4-, or 5-pyrazolyl. The preferred alicyclic and heteroalicyclic groups are 1- or 2-adamantyl, cyclohexyl, cycloheptyl, 2- or 3-tetrahydrofuranyl, 2- or 3-tetrahydrothienyl, 2- or 3-tetrahydropyranyl, 2-, 3-, or 4-piperidinyl, 3- or 4-pyrazolidinyl, 2-, 4-, or 5-thiazolidinyl, 2- or 3-piperazinyl, 2- or 3-morpholinyl, or 3- or 4-hexahydropyridazinyl.
In another aspect of the invention there is provided a method for the selective suppression of mammalian T-cell function without diminished effect on humoral immunity which comprises administering to a mammal the compound (I), whereby said compound inhibits purine nucleoside phosphorylase and thereby T-cell formation.

~''4Y YufJ'~ ' In a further aspect of the present invention there is provided a pharmaceutical composition for the selective suppression of mammalian T-cell function without diminished effect on humoral immunity which comprises an effective amount of the compound and a pharmaceutically acceptable diluent therefor.
The invention further relates to pharmaceutical compositions suitable for enteral, such as oral or rectal, transdermal and parenteral administration to mammals including man, which are useful to inhibit purine nucleoside phosphorylase activity and for the treatment of -disorders responsive thereto, comprising an effective amount of a pharmacologically active compound of the invention, alone or in combination, with one or more pharmaceutically acceptable carriers.
Preferred pharmaceutical compositions are tablets and gelatin capsules comprising the active ingredient together with a) diluents, e.g., lactose, dextrose, sucrose, mannitol, sorbitol, cellulose and/or glycine; b) lubricants, e.g., silica, talcum, stearic acid, its magnesium or calcium salt and/or polyethyleneglycol; for tablets also c) binders, e.g., magnesium aluminum silicate, starch paste, gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose and/or polyvinylpyrrolidone; if desired d) disintegrants, e.g., starches, agar, alginic acid or its sodium salt, or effervescent mixtures; and/or e) absorbents, colorants, flavors and sweeteners.
Injectable compositions are preferably aqueous isotonic solutions or suspensions, and suppositories are advantageously prepared from fatty emulsions or suspensions. Said compositions may be sterilized and/or contain adjuvants, such as preserving, stabilizing, wetting or emulsifying agents, solution promoters, salts for regulating the osmotic pressure and/or buffers. In 8 PC1'/U590/Oa756 (~y;i ... ,.:~:,,.~ .
~~;'~! :,.s~..:.d addition, they may also contain other therapeutically valuable substances. Said compositions are prepared according to conventional mixing, granulating or coating methods, respectively, and contain about 0.1 to 75%, preferably about 1 to 50%, of the active ingredient.
Suitable formulations for transdermal application include an effective amount of a compound of the invention with a carrier. Advantageous carriers include absorbable pharmacologically acceptable solvents to assist passage ZO through the skin of the host. Characteristically, transdermal devices are in the fona of a bandage comprising a backing member, a reservoir containing the compound optionally with carriers, optionally a rate controlling barrier to deliver the compound to the skin of the host at a controlled and predetermined rate over a prolonged period.
of time, and means to secure the device to the skin.
The present invention provides a method of inhibiting purine nucleoside phosphorylase activity in mammals and treating diseases and conditions responsive thereto, e.g., autoimmune disorders, rejection of transplantation, or psoriasis, which comprises administering to a mammal in need thereof an effective amount of a compound of the invention or of a pharmaceutical composition comprising a said compound in combination with one or more pharmaceutically acceptable carriers.
A further aspect of the invention relates to a method of inhibiting the phosphorolysis and metabolic breakdown of antiviral or antitumor purine nucleosides in mammals which comprises administering in conjunction therewith to a mammal in need thereof, either separately or in combination therewith, an effective purine nucleoside phosphorylase inhibiting amount of a compound of the invention or of a said compound in combination with one or more pharmaceutically acceptable carriers. More particularly, _ ~~ w~~~~
j~ ~ L.b i~Y
.Yi. ;
. ' 9' nn. '~, ~ v such relates to a method of inhibiting the phosphorolysis and metabolic breakdown of purine nucleosides known in the art, e.g., of 2'-deoxyguanosine, 2',3'-dideoxyinosine, 2',3'-dideoxyguanosine or 2',3'-dideoxyadenosine.
Furthermore, the invention thus relates to a method of potentiating the antiviral or antitumor effect of 2' or 3'-monodeoxypurine nucleosides or of 2',3'-dideoxypurine nucleosides in mammals which comprises administering in conjunction therewith to a mammal in need thereof, either separately or in combination with a said nucleoside, an effective purine nucleoside phosphorylase inhibiting amount of a compound of the invention preferably in combination with one or more pharmaceutically acceptable carriers.
More particularly, such relates to a method of enhancing or potentiating the effect of 2',3'-dideoxypurine nucleosides known in the art, e.g., of 2',3'-dideoxyinosine, 2',3'-dideoxyguanosine or 2'-3'-dideoxyadenosine for the treatment of retrovirus infections, e.g., HIV-retrovirus infections (acquired immunodeficiency syndrome, AIDS).
2o 2',3'-Dideoxypurine nucleosides are known in the art as inhibitors of HIV retrovirus infectivity and to be metabolically degraded by PNP, e.g., as described in Biochemical Pharmacolocrv 22, 3797 (1987). Such are administered at a pharmaceutically acceptable dose which is effective in inhibiting HIV-retrovirus infections.
Preferably the lowest possible effective dose is used.
The pharmaceutically acceptable effective dosage of active compound of the invention to be administered is dependent on the species of warm-blooded animal (mammal), the body weight, age and individual condition, and on the form of administration.
The pharmaceutical composition may be oral, parenteral, suppository or other form which delivers the compound into the bloodstream of a mammal to be treated. An oral form ~:..
~: "~..:1 has from about 1 to about 150 mg of the compound for an adult (5o to 70 kg) which is mixed together with pharmaceutically acceptable diluents such as lactose. In a typical capsule, 25 mg of the compound are mixed together 5 with 192 mg lactose, 80 mg modified starch and 3 mg magnesium stearate. Injectable forms of the compound are also contemplated for administration.
The present invention is also useful with other therapeutic agents. A daily dosage for a human weighing 50 10 to 70 kg of 1-50 mg/kg inhibits metabolic destruction of certain anticancer agents such as beta-2'-deoxy-6-thioguanosine and antiviral agents such as 2',3'-dideoxyinosine, an anti-AIDS drug. These types of agents are known to be susceptible to. cleavage. Upon cleavage,, the agents lose effectiveness. The compounds of the present invention are capable of reducing such cleavage.
This protection, therefore, enhances the efficacy of other chemotherapeutic agents.
One method of making the compound (I) of the present invention uses 3-substituted propionitriles as starting materials. Such starting materials can be obtained by a variety of methods that are well documented in the literature. The compound (I) is then prepared from the starting material by an adaptation of the synthetic methodology disclosed in M. I. Lim, R. S. Klein, and J. J.
Fox, J. Ora. Chem., 44, 3826 (1979); M.I. Lim, R.S. Klein, and J.J. Fox, Tetrahedron Lett.,. 21, 1013 (1980); M.I. Lim and R.S. Klein, Tetrahedron Lett., 22, 25 (1981): M. I.
Lim, W. Y. Ren, B.A. Otter, and R.S. Klein, J. Orcr. Chem., 48, 780 (1983).
A method of making the compound (II) of the present invention uses a 3-(pyridinyl)propionitrile as the starting material to make the compound (II). The appropriate 3-(pyridinyl)propionitrile can be produced by converting the t..::-: ~;
~.a,'~'a.
' ~Oh.Y f.b.~_. _ .
lI
corresponding 3-(pyridinyl)propionyl chloride to the corresponding amide by ammonolysis. with, e.g., ammonium hydroxide, which is then dehydrated to the desired nitrile by distillation with a dehydrating agent, such as POC13 or , SOC12. Alternatively, the starting material is produced by condensation of the 3-aldehyde with cyanoacetic acid followed by decarboxylation to give the corresponding substituted acrylonitrile, which is hydrogenated to give the corresponding 3-(pyridinyl)propionitrile by either catalytic hydrogenation or magnesium metal dissolving in methanol at 0 'C, such as disclosed in Profitt, J., et al., J. Org. Chem., 40, 127 (1975). The compound (II) is then prepared from the starting material by an adaptation of the synthetic methodology disclosed in M. I. Lim, R. S. Klein, and J. J. Fox, J. Org. Chem., 44, 3826 (1979); M.I. Lim,, R.S. Klein, and J.J. Fox, Tetrahedron Lett.; 21, 1013 (1980); M.I. Lim and R.S. Klein, Tetrahedron Lett., 22, 25 (1981) ; M. I. Lim, W. Y. Ren, B.A. Otter, and R.S. Klein, J. Orq. Chem., 48, 780 (1983).
Another method of making the compound of the present invention uses a known compound, i.e., cyclohexenyl-acetonitrile, as the starting material. The compound (III) of the present invention is made by reacting the starting material in an adaptation of the synthetic methodology disclosed in M. I. Lim, R. S. Klein, and J. J. Fox, J. Orcx.
Chem., 44, 3826 (1979); M.I. Lim, R.S. Klein, and J.J. Fox, Tetrahedron Lett., 21, 1013 (1980); M.I. Lim and R.S.
Klein, Tetrahedron Lett., 22, 25 (1981); M. I. Lim, W. Y.
Ren, B.A. Otter, and R.S. Klein, J. Orcr. Chem., 48, 780 (1983). Catalytic hydrogenation of either (IIIA), (IIIB), or (IIIC) yields the compound (IIID).
A method of making the compound (IV) of the present invention uses 3-substituted propionitriles as starting materials. Such starting materials can be obtained by a t ;5e-;, ((/~~ ~,..a.:~;~ .;~ ..
~~ ~'lY ~ ~~1~~~~Y'n.~

variety of methods that are well documented in the literature. The compound (IV) is then prepared from the starting material by an adaptation of the synthetic methodology disclosed in M. I. Lim, R. S. Klein, and J. J.
Fox, J. Orcr. Chem., 44, 3826 (19?9); M.I. Lim, R.S. Klein, and J.J. Fox, Tetrahedron Lett., 21, 1013 (1980); M.I. Lim and R.S. Klein, Tetrahedron Lett., ~, 25 (1981)t M. I.
Lim, W. Y. Ren, B.A. Otter, and R.S. Klein, J. org. Chem., 48, 780 (1983).
Another aspect of the present invention concerning a compound of the formula tY)p-(CH2)m-X
R'~
n provides a method of making a 2-amino compound (R1 = NH2) and intermediates thereof. The first step of the method involves reacting an optionally substituted cyclic aldehyde with cyanoacetic acid at a molar ratio of about 1/1 to 1/5 in the presence of ammonium acetate at about reflux temperature for about 10 hours to 8 days to make a 3-cyclo-substituted pentanedinitrile as an intermediate. In the second step, the 3-cyclo-pentanedinitrile is reacted with an alkyl formate such as ethyl formate and a strong base such as the metal-containing bases sodium hydride or sodium alkoxide, e.g., sodium methoxide, at a molar ratio of about 1-2/3-6/1-3 and at a temperature of about 20-f5°C for about 10 hours to 8 days to make a 3-cyclo-2-formylpentanedini-fry °.', trile as a further intermediate. The next step involves reacting the 3-cyclo-2-formylpentanedinitrile with a glycine alkyl ester hydrochloride and 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 make methyl N-((3-cyclo-2,4-dicyano)-2-butenyl]glycine as an intermediate. In the subsequent step, the methyl N-[(3-cyclo-2,4-dicyano)-2-butenyl]glycine is reacted with an alkyl chloroformate such as ethyl chloroformate and 1,5-diazabicyclo(4.3.0]non-5-ene (DBN) or 1,8-diazabicyclo [5.4.0]undec-7-ene (DSU) at a molar ratio of about 1-2/1.5-5/1.5-4 and at a temperature of about 0-50°C for about 10 hours to 10 days to make methyl 3-amino-4-(2-cyano-1-cyclo-ethyl)-1-ethyl-1H-pyrrole-1,2-dicarboxylate as an, intermediate. The next step involves reacting the methyl 3-amino-4-(2-cyano-1-cyclo-ethyl)-1-ethyl-1H-pyrrole-1,2-dicarboxylate with a base such as sodium carbonate at a molar ratio of about 2/1 to 1/5 and at about room temperature for about 10-48 hours to make methyl 3-amino-4-2~ (2-cyano-1-cyclo-ethyl)-1H-pyrrole-2-carboxylate as an intermediate. In the next step, the methyl 3-amino-4-(2-cyano-1-cyclo-ethyl)-1H-pyrrole-2-carboxylate is reacted with benzoylisothiocyanate at a molar ratio of about 2/1 to 1/2 and at about room temperature for about 30 minutes to 3 2S hours to make N-benzoyl-N--[4-(2-cyano-1-cyclo-ethyl)-2-methoxycarbonyl-1H-pyrrol-3-yl]thiourea as an intermediate.
The next step reacts the N-benzoyl-N--[4-(2-cyano-1-cyclo-ethyl)-2-methoxycarbonyl-1H-pyrrol-4-3-yl]thiourea with an alkyl halide such as methyl iodide at a molar ratio of 30 about 1/1 to 1/6 and at a temperature of about 0-30°C for about 10 minutes to 10 hours to make N-benzoyl-N--[4-(2-cyano-1-cyclo-ethyl)-2-methoxycarbonyl-1H-pyrrol-3-yl]S-methylthiourea as an intermediate. In the following step, the N-benzoyl-N'-[4-(2-cyano-1-cyclo-ethyl)-2-WO 91/06548 PCTlUS90l05756 ~.,~y IJJ.v:~'.Y~~:- ., methoxycarbonyl-1H-pyrrol-3-yl]-S=methylthiourea (about 1-2 mol) is reacted with methanolic or ethanolic ammonia at a ratio of about 1/1 to 1/20 and at a temperature of about 20-130'C for about 16-60 hours to make a mixture of a 2-amino compound of the present invention 3-cyclo-3-[2-amino-4-oxo-3H-5H-pyrrolo[3,2-d]pyrimidin-7-yl]propanenitrile and a 3-cyclo-3-[2-methylmercapto-4-oxo-3H,5H-pyrrolo[3,2-d]pyrimidin-7-yl]propanenitrile as an intermediate in making another compound of the present invention.
In a further aspect concerning the compound of the formula )p (CH2)m_!C
there is provided a method of making a 2-methoxy compound (R1 = OCH3) and intermediates thereof. The intermediate 3-cyclo-3-j2-methylmercapto-4-oxo-3H,5H-pyrrolo[3,2-d]pyrimidin-7-yl]propanenitrile is reacted with an oxidizing agent such as permanganate or hydrogen peroxide at a molar ratio of about 1/1 to 1/10 and at a temperature of about 25-120'C for about 3-48 hours to make 3-cyclo-3-[2-methylsulfonyl-4-oxo-3H,5H-pyrrolo[3,2-d]pyrimidin-7-yl]propanenitrile as an intermediate. In the next step, the 3-cyclo-3-[2-methylsulfonyl-4-oxo-3H_,5H-pyrrolo[3,2-d_]pyrimidin-7-yl]propanenitrile is reacted with a sodium alkoxide such as sodium methoxide at a molar ratio of about 1/1 to 1/10 and at_a temperature of about 25-100°C for WO 91/06548 ~ PCT/US90/05756 . ~~"~ M'~_ ~,3 ; ' about 1-48 hours to make a 2-methoxy compound of the present invention 3-cyclo-3-[2-methoxy-4-oxo-3H,5H-pyrrolo[3,2-d]pyrimidin-7-yl]propanenitrile.
In a further aspect, there is provided a method of making 5 a compound wherein R1 is hydrogen. The methyl 3-amino-4 (2-cyano-1-cyclo-ethyl)-1H-pyrrole-2-carboxylate intermediate described supra is reacted with dimethylformamide dimethyl acetal at a molar ratio of about 1/1 to 1/4 and at a temperature of about 25-100'C for about 10 1-10 days to make methyl 4-(2-cyano-1-cyclo-ethyl)-3-[N-(dimethylaminomethylene)amino]-1H-pyrrole-2-carboxylate as an intermediate. The next step involves reacting the methyl 4-(2-cyano-1-cyclo-ethyl)-3-[N-(dimethylamino-methylene)amino]-1H-pyrrole-2-carboxylate with methanolic 15 or ethanolic ammonia at a molar ratio of about 1/1 to 1/20.
and at a temperature of about 20-130'C for about 10-68 hours to make the compound of the present invention 3-cyclo-3-[4-oxo-3H,5H-pyrrolo[3,2-d]pyrimidin-7-yl]propanenitrile.
As will be apparent to the skilled artisan, variations of the aforesaid procedures are useful in making the variety of compounds of the present invention without departing from the spirit thereof. For example, compounds having different values for "n" and "m" in accordance with the formula of the present invention are obtained by either stepping up or stepping down the series by the necessary number of carbon atoms in accordance with known procedures.
Also, reactions involving some intermediates require protection of nitrogen or oxygen atoms on the intermediates 30, using known procedures.
In order to more fully describe the present invention the following non-limiting examples are provided. In the examples all parts and percentages are by weight unless WO 91/06548 PC'T/US90/05756 ': ) indicated otherwise. Proportions of solvent mixtures used as chromatographic eluents are by volume.

3-(3-Pyridinyl)propionitrile is prepared in this example.
A three-neck flask carrying a magnetic stir bar is fitted with, a thermometer, pressure equalizing addition funnel, and a reflux condenser carrying an argon inlet. Freshly powdered potassium hydroxide (6.6 g, 0.1 mol) and anhydrous acetonitrile (150 ml) are charged into the flask and heated at reflux while 3-pyridinecarboxaldehyde (10.7 g, 0.1 mol) in anhydrous acetonitrile (50 ml) is added dropwise over a period of about five minutes and refluxing continued for about another three minutes. The resulting hot reaction mixture is poured into an ice/water mixture (100 g), and , the resulting solution is extracted with CH2C12 (3 x 100 ml), dried with Na2S04, and evaporated to give crude 3-(3-pyridinyl)acrylonitrile, which is purified by column chromatography over silica gel using CHC13 as the eluent;
yield 3.3 g (25.6%).
Under an argon atmosphere, a stirred solution of the acrylonitrile (2.662 g, 0.02 mol) in 99% ethanol (100 ml) is treated with a drop of 4% aqueous sodium hydroxide followed by sodium borohydride (0.378 g, 0.01 mol).
Additional sodium borohydride (0.378 g) is added twice more at four-hour intervals. The mixture is stirred at room temperature overnight, diluted with water, extracted with EtOAc and dried with Na2S04. The solvent is removed under reduced pressure and the crude product obtained is chromatographed over a column of silica using chloroform/methanol (40/1) as the eluent to give 2.2 g (84.6%) of the product as a colorless oil.

3-(3-Pyridinyl)propionitrile of. Example 1 is further treated in the synthesis of the present invention. Under s r~
a 17 ..
an atmosphere of dry N2, a mixture of 3-(3-pyridinyl)propionitrile (0.661 g, 5.0 mmole), sodium hydride (0.240 g, 10.0 mmole), and ethyl formats (1.11 g, 15.0 mmole) in anhydrous tetrahydrofuran (20 ml) is stirred for 48 hours with protection from air and moisture.
Volatile matter is evaporated, and a solution of the solid residue in 15 ml of sold water is adjusted at 0 ° C to a pH
of 6 with cold 6N HCl. The resulting oily mixture is extracted with CHC13, and the extract is washed with water, dried using Na2S04, and evaporated to give a dark oil, which is a mixture of 2-formyl-3-(3-pyridinyl)propionitrile and the nitrile starting material. This crude product is ;sed in the next reaction without further purification.

i5 Glycine methyl ester hydrochloride (0.942 g, 7.5 mmole) and anhydrous sodium acetate (0.615-g, 7.5 mmole) are added to a solution of the crude formyl compound (0.89 g) in MeOH/H20 (4:1, 50 ml). After 24 hours, the MeOH is evaporated in vacuo, and the mixture of water and oil is extracted with CHC13. The CHC13 layer is dried (Na2S04) and evaporated to give an amber oil which is applied to a silica gel column. Elution with CHC13 gave two major bands: (1) 3-(3-pyridinyl)propionitrile (used as starting material in the previous step), and (2) the desired enamine.

Under a nitrogen atmosphere, ethyl chloroformate (0.521 g, 4.8 mmole) is added dropwise to a solution of the enamine of Example 3 (0.513 g, 3.2 mmole) and 1,5-diazabicyclo[4.3.0)non-5-ene (nBN, 1.37 g, 11.1 mmole) in dry CH2C12 (15 ml) with external cooling in an ice bath.
After stirring at 0 °C for one hour, the solution is allowed to a stand at room temperature overnight. After checking progress by TLC, additional ClCO2Et (0.1 ml) and ('~":''.;

DBN (1.o ml) are added to complete the conversion, and the solution is allowed to stand for 24 hours. Volatile matter is evaporated in vacuo and the viscous residue is purified on a short silica gel column (whose main purpose is to remove the less-mobile DBN) to give an N-blocked pyrrole;
which is used for the next step without further purification.

To a solution of the N-blocked pyrrole of Example 4 (0.635. g, 2.0 mmole) in MeOH (50 ml) is added solid Na2C03 (0.212 g, 2.0 mmole), and the reaction mixture is stirred at room temperature for 48 hr with separation of the resultant deblocked pyrrole. The mixture is evaporated to dryness, and the residue is triturated thoroughly with H20 .
(25 ml) to dissolve inorganics and extracted with CHC13 (3 x 100 ml). The extract is dried (Na2S04) and evaporated to give a viscous gum that crystallized upon drying in vacuo for use as an intermediate without further purification.
More extensive purification can, however, be effected by using either column chromatography employing silica gel/CHC13 or recrystallization from toluene/cyclohexane (1:3).

Benzoyl isothiocyanate (0.232 g, 1.42 mmole) is added dropwise to a solution of the unblocked pyrrole of Example 5 (0.290 g, 1.18 mmole) in dry CH2C12 (100 ml). After 1 h at room temperature, solution is evaporated, and the gummy residue is stirred in Et20/cyclohexane (1:1, 20 ml). The resulting suspension of yellow solid is filtered under N2 pressure, and the thioureido product is dried in vacuo over P205.

Methyl iodide (0.228 g, 1.61 mmole) is added to a solution of the thioureido product of Example 6 (0.383 g, r., ~~a~~~~i~C ..

0.94 mmole) and DBN (0.140 g, 1.12 mmole) in dry CH2C12 (10 ml) at 0 'C. The solution is stirred at 0 °C for 15 min., at ambient temperature for 1 h, and then evaporated in vacuo. A solution of the residue in CHC13 is chromatographed on a silica gel column with CHC13/methanol (97:3) as eluent to give homogeneous fractions of the methylthio intermediate compound.

A solution of the methylthio compound of Example ?
(0.358 g, 0.85 mmole) in 100 ml of MeOH that has been saturated with NH3 at 0 'C is 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 the desired 2-amino intermediate compound, benzamide, and a by-product that is a 2-methylthio .
derivative, as opposed to the 2-amino compound. The mixture is dissolved in methanol and the solution is evaporated with silica gel (about 5 g). The mixture is then carefully layered onto the top of a silica-gel chromatography column, which is then eluted with CHC13/MeOH
(9:1) to give the methylthio by-product and the desired 2 amino-7-(3-pyridinylmethyl)-3#~,5i~-pyrrolo(3,2-d_]pyrimidin 4-one intermediate. Further purification is obtained by recrystallization from boiling isopropyl acetate in a Soxhlet apparatus.
A solution of the pyridinylmethyl intermediate in O.1N
HCl is hydrogenated with a platinum catalyst at 60 lb/in2 H2 pressure. The catalyst is generated by brief .
hydrogenation of Pt02 in O.1N HC1. When the reaction is complete, the catalyst is removed by filtration under N2 pressure, and the filtrate is evaporated. A solution of the residue in the minimum amount of ethanol is diluted slowly with a large amount of Eto2, and the hydrochloride ~~:~'~ ~~.~3 salt is obtained as a white hygroscopic solid of the compound (IC).

The compound (IC) of Example 8 is tested foz enzyme 5 inhibition activity. A purine nucleoside phosphorylase (PNP) enzyme assay is performed in which PNP activity (IC50) for the compound is observed, which is determined radiochemically by measuring the formation of [14C]
hypoxanthine from (14CJ-inosine (see Biomedicine, 33, 39 10 (1980)) using calf spleen as the enzyme source.

The following compounds of the present invention are prepared . that are 2-amino-7-(R)-3H_,5H-pyrrolo[3,2-d_]-pyrimidin-4-ones wherein R is -CH2R1 in which the R1 group , 15 is as follows:
Example 10 R1 = 2-methyl-3-piperidinyl Example 11 R1 = 2-chloro-3-piperidinyl Example 12 R1 = 2-trifluoromethyl-3-piperidinyl Example 13 R1 = 2-methoxy-3-piperidinyl 20 Example 14 R1 = 2-fluoro-3-piperidinyl The compounds are prepared following the procedures set forth in Examples 1-8 using the appropriate 3-(substituted 3-pyridinyl)-propionitriles as starting materials.

3-(2-Pyridinyl)propionitrile is prepared in this example using the procedure of V. Boekelheide, et al., J. Am. Chem.
Soc., 75, 3243 (1953). A solution of potassium cyanide (83.74 g) in water (160 ml) is added to a solution of freshly distilled 2-vinylpyridine (67.59 g) in acetic anhydride (131.30 g) with the rate of addition adjusted to maintain gentle refluxing. 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 is adjusted to pH 8 with a saturated 21 ~o~~ ~ ~~ r-'~'N-~
Na2C03 solution. The mixture is extracted with CHC13 (4 x 150 ml), washed with water, dried with Na2S04, and evaporated to a dark viscous oil. The oil is fractionally distilled in vacuo through a vigreaux column. After a small forerun containing 2-vinylpyridine (0.35 g), the desired 3-(2-pyridinyl)propionitrile is collected as a clear, fluorescent yellow-green, viscous oil: yield 59.8 g;
70.4%; by 86°C at 1.0 mm Hg.

The following compounds of the present invention are prepared that are 2-amino-7-(R)-3H,5H-pyrrolo[3,2-d]
pyrimidin-4-ones wherein R is -CH2R1 in which the R1 group is as follows:
Example 16 R1 = 2-piperidinyl Example 17 R1 = 4-piperidinyl Example 18 R1 = 3-trifluoromethyl-4-piperidinyl Example 19 R1 = 3-methoxy-2-piperidinyl Example 20 R1 = 3-fluoro-4-piperidinyl The compounds are prepared following the procedures set 2o forth in Examples 1-8 and 15 using the appropriate 3-(2- or 4-pyridinyl)-propionitriles as starting materials.

A pharmaceutical composition for intraperitoneal injection is prepared for testing the compound (IC). An intraperitoneal injection solution containing the compound of Example 8 is dissolved in an aqueous carrier that contains ten percent DMSO.

The compound (IC) is intraperitoneally injected into Lewis Rats via the test composition of Example 21 to provide 30 mg of the compound (IC), with an injection given twice per day. Controls are used, which receive only the vehicle. At specific times after administration, the animals are sacrificed and plasma samples are prepared.

l~fe~'r~ :e~ y4J ~~ i The plasma is extracted with cold 0.5 N HC104 and neutralized with solid NH4HC03. After removal of perchlorate salts, the extract is subjected to HPLC on a reversed phase column (Spherisorb ODSI). A significant increase in plasma inosine,is observed in the plasma taken from animals receiving the compound (IC).

Compounds prepared as in Examples 10-20 are each made into a pharmaceutical formulation in accordance with the preparation of Example 21 and the resultant injectable solutions are tested in accordance with the procedure of Example 22. A significant increase in plasma inosine is observed in the plasma taken from animals receiving the compounds of the present invention.

3-(2-Furanyl)propionitrile is prepared in this example.
In a three-neck flask fitted with a condenser and drying tube, magnesium turnings (20 g) are added to a solution of 3-(2-furanyl)acrylonitrile (67.2 g) in dry methanol (2 1).
Additional magnesium is added in parts as the reaction rate would allow until a total of 145 g has been added. After about five hours, the solvent is evaporated until the contents set to a solid paste, which is adjusted to a pH of about 6.5 With 6N HC1 with cooling. The mixture is extracted with several portions of CHC13, dried with Na2S04, and concentrated to a dark oil. Distillation in vacuo through a short Vigreaux column gives the propionitrile as a colorless oil; yield 49.5 g (72%); by 46.0-46.5°C at 0.5 mm.

3-(2-Furanyl)propionitrile of Example 34 is further treated in the synthesis of the present invention. Under an atmosphere of dry N2, the propionitrile (48.7 g), sodium hydride (10.28 g), ethyl formate (32.74 g), and anhydrous tetrahydrofuran (200 ml) are stirred at room temperature with protection from moisture for about 18 hours. The ~_' volatile matter i~'Hthen evaporated, the resulting yellow solid is dissolved in about 20 ml of cold water with ice s bath cooling, and the solution is adjusted to a pH of 6.0 with cold 6N HCl. The resulting heavy oil precipitate is extracted into CHC13, and the extract is washed with water, dried with Na2S04, and evaporated to give a thin oil which contains the crude formyl compound; yield 53.06 g.

Glycine methyl ester hydrochloride (67.05 g) and anhydrous sodium acetate (43.79 g) are added to a solution of the crude formyl compound of the previous example (53.05 g) in MeOH/H20 (4:1, 1500 ml). After 24 hours, the.
MeOH is evaporated in vacuo, and the mixture of water and oil is extracted with CHC13. The CHC13 layer is dried (Na2S04) and evaporated to give an amber oil (59.1 g).

Under a nitrogen atmosphere, ethyl chloroformate (43.68 g) is added dropwise to a solution of the amber oil of Example 36 (59.1 g) and DBN (133 g) in dry CH2C12 (400 ml) with external cooling in an ice bath. After stirring at 0 ° C for one hour, the solution is allowed to stand at room temperature overnight. After checking progress by TLC, additional C1C02Et and DBN are added to complete the conversion, and the solution is allowed to stand for 24 hours. Volatile matter is evaporated in vacuo, the viscous residue purified on a silica gel column (whose main purpose is to remove the less-mobile DBN) to give an N-blocked pyrrole, a corresponding pyrrole without the ethoxycarbonyl blocking group on the pyrrole nitrogen, and the starting propionitrile.
x ~.~,,,~.:1 ~~? r m~~~ 2~~

To a solution of the N-blocked pyrrole of Example 37 (11.66 g) in MeOH (200 ml) is added solid Na2CO3 (4.23 g), and the reaction mixture is stirred at room temperature for 24 hr with separation of the resultant deblocked pyrrole.
The mixture is evaporated to dryness, and the residue is triturated thoroughly With H20 (200 ml) to dissolve inorganics and extracted with CHC13 (3 x 200 ml). The extract is dried (Na2S04) and evaporated to give a viscous gum that crystallized upon drying in vacuo for use as an intermediate without further purification. More extensive purification can, however, be effected by using either column chromatography employing silica gel/CHC13 or recrystallization from toluene/cyclohexane (1:3).

Benzoyl isothiocyanate (4.58 g) is added dropwise to a solution of the unblocked pyrrole of Example 38 (5.15 g) in dry CH2C12 (75 ml). After 1 h at room temperature, solution is evaporated, and the gummy residue is dissolved in Et20 (100 ml) with almost immediate separation of the crystalline solid, which is collected by filtration. The Et20 filtrate is heated to boiling and diluted with an equal volume of warm cyclohexane. On cooling slowly the solution gives additional thioureido product.

Methyl iodide (8.70 g) is added to a solution of the thioureido product of Example 39 (10.68 g) and DBN (4.15 g) in dry CH2C12 (250 ml) at 0 °C. The solution is stirred at 0 °C for 15 min., at ambient temperature .for 1 h, and then evaporated in vacuo. A solution of the residue in CHC13 is chromatographed on a silica gel column with CHC13 as eluent to give homogeneous fractions of the methylthio intermediate compound.

~, rah ,: ~~: ti.~:~.r~'~.~
. ~.a A solution of the methylthio compound of Example 40 (0.80 g, 2.0 mmole) in 25 ml of MeOH that has been saturated with NH3 at 0 °C is heated at 90-95 °C for 24 5 hours in a glass-lined stainless steel bomb. The contents of the chilled bomb axe evaporated in vacuo to give a mixture of the 2-amino compound, benzamide and a by-product that is a 2-methylthio derivative, as opposed to the 2-amino compound. The mixture is stirred vigorously for 10 several minutes with 30 ml of 2:1 Et20/cyclohexane, and the insoluble white solid is filtered off and washed with Et20.
The filtrate contained most of the benzamide and 2-methylthio components. A solution of the Et20/cyclohexane-insoluble solid (0.425 g) in MeOH is evaporated with appr.
15 10 g of silica gel. The powdered residue is layered evenly.
onto the top of a silica gel column, which is then eluted with CHC13/MeOH/HOAc (95:5:1) to give the 2-methylthio by-product and the desired furanyl intermediate. The intermediate is recrystallized by extraction into boiling 20 isopropyl acetate in a Soxhlet apparatus. The white crystals are collected in three crops and dried in vacuo over P205 at 110°C for 7h.
A solution of the intermediate (116 g, 0.5 mmole) in methanol (50 ml) is hydrogenated with 30% palladium-on 25 charcoal (40 mg) at 62 lb/in2 H2 pressure for about 36 hours. The catalyst is removed by filtration under N2 pressure, and the filtrate is evaporated and reevaporated with toluene. The solid residue is recrystallized by extraction into boiling isopropyl acetate in a soxhlet apparatus. The 2-tetrahydrofuranyl compound (IE) is obtained as a white crystalline solid, which is dried in vacuo over P205 at 110°C for about six hours; yield 73 mg (61.8%): m.p. 284-286°C dec.

t,'T~:.' .....

The product of Example 41 is tested for enzyme inhibition activity according to the procedure of Example 9, and PNP
activity (IC50) for the compound is observed.

A pharmaceutical composition for intraperitoneal injection is prepared for testing the compound (IE). An intraperitoneal injection solution containing the compound of Example 41 is dissolved in an aqueous carrier that l0 contains ten percent DMSO.

Using the procedure of Example 16, the compound (IE) is intraperitoneally injected into Lewis Rats via the test composition of Example 43 to provide 30 mg of the compound, (IE) and the results are analyzed compared to controls. A
significant increase in plasma inosine is observed in the plasma taken from animals receiving the compound (IE).

The following compounds of the present invention are prepared that are 2-amino-7-(R)-3F~,5H_-pyrrolo[3,2-d_]
pyrimidin-4-ones wherein R is -CH2-R1 in which the R1 group is as follows:
Example 45 R1 = 3-tetrahydrofuranyl Example 46 R1 = 3-chloro-2-tetrahydrofuranyl Example 47 R1 =~ 3-trifluoromethyl-2-tetrahydrofuranyl Example 48 R1 = 3-methoxy-3-tetrahydrofuranyl Example 49 R1 = 3-fluoro-2-tetrahydrofuranyl The compounds are prepared following the procedures set forth in Examples 34-41 using the appropriate 3-(furanyl) acrylonitriles as starting materials.

Under a nitrogen atmosphere, a solution of the tetrahydrofuranyl compound (IE) (50o mg) obtained from Example 41 and a crystal of phenol in 2N HBr (20 ml) is WO 91/06548 PCT/US90l05756 ~, r.~" j .~y , , .
~..' .f :.., ~.: .,~

stirred for 18 hours at 40'C. The solvent is evaporated vacuo at low temperature, and the residue is washed with a few ml of Et20 by decantation to remove any tribromophenol.
A suspension of the residue in H2o is adjusted to pH 7 with 1N NaOH to give the bromoalcohol as its free base. The solid is collected, washed with cold water, and dried at room temperature.
A solution of the bromoalcohol in anhydrous dimethyl-acetamide is chilled in an ice bath and treated with an equimolar portion of PBr3. The solution is allowed to stir at room temperature for one hour, then the dimethylacetamide is evaporated in vacuo at low temperature using an air pump and dry-ice trap. A suspension of the residue in ice/water is adjusted to pH 7 using 1N NaOH to give the crude reaction product containing the 1,4-dibromo compound. After filtration, washing with cold water, and drying at room temperature, the resulting product is refluxed with sodium sulfide in 50% ethanol/water, or alternatively warmed with Na2S in N,N-dimethylacetamide solution; the solvent removed under vacuum, the residue washed with water to remove NaBr, and the pH adjusted to about 7 to precipitate the 2-tetrahydrothienyl compound (IG) .

The compound of Example 50 is tested for enzyme inhibition activity as in Example 9, and PNP activity (IC50) for the compound is observed.

A pharmaceutical composition for intraperitoneal injection is prepared for testing the compound (IG). An intraperitoneal injection solution is made containing the compound of Example 50 is dissolved in an aqueous carrier that contains ten percent DMSO.

Using the procedure of Example 16, the compound (IG) is intraperitoneally injected into Lewis Rats via the test composition of Example 52 to provide 30 mg of the compound (IG) and the results are analyzed compared to controls. A
significant increase in plasma inosine is observed in the plasma taken from animals receiving the compound (IG).

The following compounds of the present invention are prepared that are 2-amino-7-(R)-3$,5H-pyrrolo[3,2-d_]
pyrimidin-4-ones wherein R is -CH2-R1 in which the R1 group is as follows:
Example 54 R1 = 3-tetrahydrothienyl Example 55 R1 = 3-chloro-2-tetrahydrothienyl Example 56 R1 = 3-trifluoromethyl-2-tetrahydrothienyl Example 57 R1 = 3-methoxy-2-tetrahydrothienyl Example 58 R1 = 3-fluoro-2-tetrahydrothienyl The compounds are prepared following the procedures set forth in Examples 34-41 and 50 using the appropriate 3 (furanyl)-acrylonitriles as starting materials.

Using the procedure of Example 1, 3-(2-pyrrolyl)-propionitrile is prepared using pyrrole-2-carboxaldehyde as the starting material. Following Examples 2-8, the intermediate 2-amino-7-(2-pyrrolylmethyl)-3H,5Fi-pyrrolo[3,2-d]pyrimidin-4-one is prepared from the propionitrile, from which the 2-pyrrolidinyl compound (II) is obtained by reduction of the intermediate.

Using the procedure of Example 1, 3-(3-pyrrolyl)-propionitrile is 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 prepared from the propionitrile, WO 91/0648 PCTlUS90l05756 .~_~.
29 ~~~ ~s~M.J:.i~:Y~'~
from which the 3-pyrrolidinyl compound (IJ) is obtained by reduction of the intermediate.

Using the procedure of Example 1, 3-(2-pyranyl) propionitrile is prepared using 2-pyrancarboxaldehyde as the starting material. Following Examples 2-8, the intermediate 2-amino-7-(2-pyranylmethyl)-3I~,5~i-pyrrolo-[3,2-_d]pyrimidin-4-one is prepared from the propionitrile, from which the 2-tetrahydro-pyranyl compound (IL) is obtained by reduction of the intermediate.

Using the procedure of Example 1, 3-(3-pyranyl) propionitrile is prepared using 3-pyrancarboxaldehyde as the starting material. Following Examples 2-8, the intermediate 2-amino-7-(3-pyranylmethyl)-3H,5H-pyrrolo[3,2-dJpyrimidin-4-one is prepared' from the propionitrile, from which the 3-tetrahydro-pyranyl compound (IM) is obtained by reduction of the intermediate.

Using the procedure of Example 1, 3-(4-pyranyl) propionitrile is 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 prepared from the propionitrile, from which the 4-tetrahydro-pyranyl compound (IN) is obtained by reduction of the intermediate.

A solution of the methylthio compound of Example 7 (0.358 g, 0.85 mmole) in 100 ml of MeOH that has been saturated with NH3 at 0 °C is 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 the desired 2-amino intermediate compound, benzamide, and a by-product that is a 2-methylthio WO 91/06548 PCTlUS90/05756 ~~,.i ~o~ti.i~ :aas,.~~y , , derivative, as opposed to the 2-amino compound. The mixture is dissolved in methanol and the solution is evaporated with silica gel (about 5 g). The mixture is then carefully layered onto the top of a silica-gel 5 chromatography column, which is then eluted with CHC13/MeOH
(9:1) to give the methylthio by-product and the desired 2 amino-7-(3-pyridinylmethyl)-3I~,5H_-pyrrolo[3,2-d_]pyrimidin 4-one, compound (IIA). Further purification is obtained by recrystallization from boiling isopropyl acetate in a 10 Soxhlet apparatus.

The compound (IIA) of Example 64 is tested for enzyme inhibition activity. A purine nucleoside phosphorylase (PNP) enzyme assay is performed in which PNP activity 15 (IC50) for the compound is observed, which is determined radiochemically by measuring the formation of [14C]-hypoxanthine from [14C]-inosine (see Biomedicine, 33, 39 (1980) using calf spleen as the enzyme source.

2~ The following compounds of the present invention are prepared that are 2-amino-7-(R)-3H_,5H_-pyrrolo[3,2-d]
pyrimidin-4-ones wherein R is -CH2-R1 in which the R1 group is as follows:
Example 66 R1 = 2-pyridinyl 25 Example 67 R1 = 4-pyridinyl Example 68 R1 = 3-chloro-2-pyridinyl The compounds are prepared following the procedures set forth in Examples 1-7 and 64 using the appropriate 3-(pyridinyl)-propionitriles as starting materials.

A pharmaceutical composition for intraperitoneal injection is prepared for testing the compound (IIA). An intraperitoneal injection solution containing the compound r~.
f...
~~',, s~,~~~''3 of Example 64 is dissolved in an aqueous carrier that contains ten percent DMSO.

The compound (IIA) is intraperitoneally injected into Lewis Rats via the test composition of Example 69 to provide 30 mg of the compound (IIA), with an injection given twice per day. Controls are used, which receive only the vehicle. At specific times after administration, the animals are sacrificed and plasma samples are prepared.
The plasma is extracted with cold 0.5 N HC104 and neutralized With solid NH4HCOg. After removal of perchlorate salts, the extract is subjected to HPLC on a reversed phase column (Spherisorb ODSI). A significant increase in plasma inosine is observed in the plasma taken .
from animals receiving the compound (TIA).

Compounds prepared as in Examples 66-68 are each made into a pharmaceutical formulation in accordance with the preparation of Example 69 and the resultant injectable solutions are tested in accordance with the procedure of Example 70. A significant increase in plasma inasine is observed in the plasma taken from animals receiving the compounds of the present invention.

1-Cyclohexenylacetonitrile is treated in the synthesis of the present invention. Under an atmosphere of dry N2, a solution of 1-cyclohexenylacetonitrile (9.2 g: 75.92 mmole) in anhydrous tetrahydrofuran (THF, l0 ml) is added to a stirred mixture of sodium hydride (3.18 g; 132.86 mmole) and ethylformate (30.14 g; 406.93 mmole) in 50 ml THF, and the resulting mixture is stirred at room temperature for about 18 hours. Volatile matter is evaporated in vacuo at room temperature. Water (30 ml) is added to the residue at 0'C, and the solution adjusted to a pH of 6 by dropwise ,r..~,, iv.';
~~; t,.'~""'~ r' ~~

addition of 6N HC1. The resulting precipitate of heavy oil is extracted into CHC13. The extract is washed with water and dried with Na2SO4, and the resulting organic layer evaporated to give a crude formyl compound as a red-brown oil (9.6 g).

Glycine methyl ester hydrochloride (16.68 g, 132.85 mmole) and anhydrous sodium acetate (10.89 g, 132.85 mmole) are added to a solution of the crude formyl compound (9.6 g) of Example 74 without further purification in MeOH/H2o (4:1, 150 ml). After 24 hours, the MeOH is evaporated in vacuo, and the mixture of water and oil is extracted with CHC13. The CHC13 layer is dried (Na2S04) and evaporated to give an amber oil which is applied to a silica gel column.
_.. ~lution with CHC13 gave the desired enamine: yield 4.5 g.

Under a nitrogen atmosphere, ethyl chloroformate (3.04 g;
28.06 mmole) is added dropwise to a solution of the enamine of Example 75 (4.12 g, 18.70 mmole) and 1,5-di-azabicyclo[4.3.0]-non-5-ene (DBN, 6.96 g, 56.11 mmole) in dry CH2C12 (100 ml) with external cooling in an ice bath.
After stirring at 0 'C for one hour, the solution is allowed to stand at room temperature overnight. After checking progress by TLC, additional C1C02Et (0.5 ml) and DBN (3.0 ml) are added to complete the conversion, and the solution is allowed to stand for 24 hours. Volatile matter is evaporated in vacuo, the viscous residue purified on a short silica gel column (whose main purpose is to remove the less-mobile DBN) to give an N-blocked pyrrole, which is used for the next step without further purification.

To a solution of the N-blocked pyrrole of Example 76 (3.0 g, 10.26 mmole) in MeOH (100 ml) is added solid Na2C03 (2.71 g, 25.65 mmole), and the reaction mixture is stirred ...,.
.:, at room temperature for 48 hr with separation of the resultant deblocked pyrrole. The mixture is evaporated to dryness, and the residue is triturated thoroughly with H2o (50 ml) to dissolve inorganics and extracted with CHC13 (3 x 50 ml). The extract is dried (Na2so4) and evaporated to give a viscous gum, which is purified on a silica gel column using CHC13 as the eluent; yield 2.04 g; m.p. 125°C.

Benzoyl isothiocyanate (0.76g, 4.65 mmole) is added dropwise to a solution of the unblocked pyrrole of Example 77 (0.91 g, 4.13 mmole) in dry CH2C12 (30 ml). After 1 h at room temperature, the solution is evaporated, and the gummy residue is triturated with methanol to give a thioureido product: yield 0.70 g~ m.p. 170°C.

Methyl iodide (0.678 g, 4.78 mmole) is added to a solution of the thioureido product of Example 78 (0.630 g, 1.64 mmole) and DBN (0.230 g, 1.85 mmole) in dry CH2C12 (50 ml) at 0 °C. The solution .is stirred at 0 °C for 15 min., at ambient temperature for 1 h, and then evaporated in vacuo. A solution of the residue in CHC13 is chromatographed on a silica gel column with CHC13 as eluent to give homogeneous fractions of the methylthio intermediate; yield 0.7 g.

A solution of the methylthio compound of Example 79 (0.70 g, 1.76 mmole) in 50 ml of MeOH that has been saturated with NH3 at 0 °C is 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 the compound (IIIA), benzamide and a by-product that is a 2-methylthio derivative, as opposed to the 2-amino compound (IIIA). The mixture is stirred vigorously for several minutes with appr. 50 ml of Et20, and the WO 91/06548 PCT/US90f05756 ~'~. ;1 insoluble white solid is filtered off and washed with Et20.
The filtrate contained most of the benzamide and 2-methylthio components. A solution of the Et20-insoluble solid (0.342 g) in MeOH is evaporated with appr. to g of silica gel. The powdered residue is layered evenly onto the top of a silica gel column, which is then eluted with CHC13/MeOH/HOAc (95:5:1) to give the 2-methylthio by-product and the desired 2-amino product (IIIA). (IIIA) is recrystallized by extraction into boiling isopropyl acetate in a Soxhlet apparatus. The white crystals are collected in three crops and dried in vacuo over P2o5 at 110 ' C for 7h; yield 44%: mp 280 'C dec., anal. calcd. for C12H14N40'0.6H20: C, 59.78; H. 6.35; N, 23.23. Found: C, 59.98; H, 6.46; N, 23.15.

The compound of Example 80 is tested for enzyme-inhibition activity. A purine nucleoside phosphorylase (PNP) enzyme assay is performed in which PNP activity for the compound is determined radiachemically by measuring the formation of [14C]-hypoxanthine from [14C]-inosine (see Biomedicine, 33, 39 (1980) using calf spleen as the enzyme source. At 1 mM phosphate the IC50 is 1.9 ~.M, and at 50 mM
phosphate the IC50 is 19 ~M.

Following the procedures set forth in Examples 74-80, compounds (IIIB) and (IIIC) are made using 2- and 3-cyclohexenyl-acetonitrile, respectively, as starting materials. The compounds are tested as in Example 81 and significant enzyme-inhibition activity is observed.

The following compounds of the present invention are prepared that are 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 r'r' ;.
.. w "~" p:' 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 5 The compounds are prepared following the procedures set forth in Examples 74-81 using the appropriate substituted cyclohexenyl acetonitriles as starting materials.

A pharmaceutical composition for intraperitoneal 10 injection is prepared for testing the compound (IIIA). An intraperitoneal injection solution containing the compound (IIIA) is dissolved in an aqueous carrier that contains ten percent DMSO.

15 The compound (IIIA) is intraperitoneally injected into Lewis Rats via the test composition of Example 88 to provide 30 mg of the compound (IIIA), with an injection given twice per day. Controls are used, which receive only _ the vehicle. At specific times after administration, the 20 animals are sacrificed and plasma samples are prepared.
The plasma is extracted with cold 0.5 N HC104 and neutralized with solid NH4HC03. After removal of perchlorate salts, the extract is subjected to HPLC on a reversed, phase column (Spherisorb. ODSI). A significant 25 increase in plasma inosine is observed in the plasma taken from animals receiving the compound (IIIA).

Compounds grepared as in Examples 83-87 are each made into a pharmaceutical formulation in accordance with the 30 preparation of Example 88 and the resultant injectable solutions are tested in accordance with the procedure of Example 89. A significant increase in plasma inosine is observed in the plasma taken from animals receiving the compounds of the present invention.

~~ 1~,~ ~~

The compound (IIID) is prepared using 2-amino-7-(1-cyclohexenyl)-3H,5H-pyrrolo[3,2-d]pyrimidin-4-one as an intermediate. A solution of the intermediate (0.2 g; 0.86 mmole) in ethanol (50 ml) is hydrogenated with 10% Pd-C (50 mg) at 45 lb/in2 for 16 h and filtered hot through Celite.
The filtrate is evaporated to dryness, and the residue is crystallized from hot ethanol to give the compound (IIID);
yield 157 mg (78%), mp >300 'C dec. anal. calcd. for C12H16N40'0. EtOH: C, 61.80; H, 7.10; N, 23.51. Found:
C, 61.95; H, 7.43: N, 23.56.

The compound (IIID) prepared in Example 95 is tested for enzyme-inhibition activity as in Example 81. At 1 mM.
phosphate the IC50 is 1.3 uM, and at 50 mM phosphate the IC50 is 145 ACM.

3-(2-Adamantyl)propionitrile is prepared in this example using a modification of the procedure of M. Ohno, et al. , J. Org. Chem. 53, 1285 (1988). A Solution of 2-bromoadmantane (20 g;,92.96 mmole); Bu3SnH (32.46 g; 111.5 mmole), acrylonitrile (9,86 g; 185.92 mmole), and AIBN (740 mg) in toluene (280 ml) is stirred at reflux temperature for 3 h. The reaction mixture is washed with ammonia water (0.4 M, 500 ml), the organic layer is washed with H20 and dried aver MgS04 and evaporated. The residue is distilled between 110-118'C (and about 0.2 mmHg); fractions are combined to give a crude sample of contaminated 3-(2-adamantyl)propionitrile with tin complexes, which is purified on silica gel column with hexanes; followed by hexanes/ethylacetate 97:3 and hexanes/ethylacetate 95:5, yield 9.4 g (53.4%); mp semi-solid.

37 ~~,~;~~~,,~~3 3-(2-Adamantyl)propionitrile of Example 97 is further treated in the synthesis of the present invention. Under an atmosphere of dry N2, a mixture of 3-(2-adamantyl) propionitrile (7.0 g, 36.99 mmole), sodium hydride (1.7 g, 73.95 mmole), and anhydrous tetrahydrofuran (?5 ml) is heated at 52'C in a water bath for 15 min., and a solution of ethyl formate (13.69 g, 184.89 mmole) in THF (100 ml) is added over a period of 45 min. After two hours at 50 - 55 °C, a second portion of NaH.(0.8 g) and HC02Et (7.5 ml) are added, and the reaction mixture is stirred for about two days. A third portion of HC02Et (7.5 ml) and NaH (0.8 g) are added and left at room temperature for about 24 hours (unreacted nitrile is inert in the next step and can be recovered at the first purification step). The thick paste is stirred overnight and allowed to cool to room temperature. Volatile matter is evaporated under reduced pressure, and the residual pale yellow crust is dissolved in the minimum volume of cold water (appr. 150 ml) at 0 °C.
The solution is adjusted to pH 6.0 by addition of 6N HC1 and extracted with CHC13 (3 x 100 ml). The extract is washed with H20, dried over Na2SO4, and evaporated in vacuo to a thick amber oil. This crude product is used in the next reaction without further purification.

Glycine methyl ester hydrochloride (6.96 g, 55.46 mmole) and anhydrous sodium acetate (4.55 g, 55.46 mmole) are added to a solution of the crude formyl compound (8.0 g) in MeOH/H20 (4:1, 500 ml). After 24 hours, the MeOH is evaporated in vacuo, and the mixture of water and oil is extracted with CHC13. The CHC13 layer is dried (Na2S04) and evaporated to give an amber oil which is applied to a silica gel column. Elution with CHC13 gave two major bands: (1) 3-(2-adamantyl)-propionitrile (used as star'cing ~,~;~~?~ ~~=3 material in the previous step), and (2) the desired enamine: yield 6 g.

Under a nitrogen atmosphere, ethyl chloroformate (2.82 g, 26.0 mmole) is added dropwise to a solution of the enamine of Example 99 (5.0 g, 17.34 mmole) and 1,5-diazabicyclo [4.3.0]non-5-ene ("DBN,~~ 6.46 g, 52.0 mmole) in dry CH2C12 (50 ml) with external cooling in an ice bath. After stirring at 0 'C for one hour, the solution is allowed to a stand at room temperature overnight. After checking progress by TLC, additional C1C02Et (1.81 ml) and DBN
(3.23 g) are added to complete the conversion, and the solution is allowed to stand for 24 hours. Volatile matter is evaporated in vacuo, the viscous residue purified on a short silica gel column (whose main purpose is to remove.
the less-mobile DBN) to give an N-blocked pyrrole, which is used for the next step without further purification.

To a solution of the crude N-blocked pyrrole of Example 100 (6.0 g, 16.59 mmole) in MeOH (100 ml) is added solid Na2C03 (4.39 g, 41.49 mmole), and the reaction mixture is stirred at room temperature for 48 hr with separation of the resultant deblocked pyrrole. The mixture is evaporated to dryness, and the residue is triturated thoroughly with H20 (50 ml) to dissolve inorganics and extracted with CHC13 (3 x 100 ml). The extract is dried (Na2S04) and evaporated to give a viscous gum, which crystallized by triturating with ether: yield 4 g; m.p. 162-163°C.

Benzoyl isothiocyanate (1.22 g, 7.47 mmole) is added dropwise to a solution of the unblocked pyrrole of Example 101 (1.91 g, 6.62 mmole) in dry CH2C12 (50 m1). After 1 h at room temperature, solution is evaporated, and the gummy residue is dissolved in Et20 (100 ml) with almost immediate ,~'aF~"a'~ '?~
..'~~.sa. r-.r~

separation of the crystalline solid. The Et20 filtrate is heated to boiling and diluted with an equal volume of warm cyclohexane. On cooling slowly the solution gives additional thioureido product: yiled 2.81 g (95%); m.p.
193-194°C.

Methyl iodide (2.46 g, 17.39 mmole) is added to a solution of the thioureido product of Example 102 (2.7 g, 5.98 mmole) and DBN (0.82 g, 6.57 mmole) in dry CH2C12 (50 ml) at 0 °C. The solution is stirred at 0 °C for 15 min., at ambient temperature for 1 h, and then evaporated in vacuo to give a crude sample of the methythio intermediate compound; yield 2.78 g (crude).

A solution of the methylthio compound of Example 103 (2.78 g, 5.18 mmole) in 150 ml of MeOH that has been saturated with NH3 at 0 °C is 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 the compound (IVA), benzamide and a by-product that is a 2-methylthio derivative, as opposed to the 2-amino compound (IVA). The mixture is stirred vigorously for several minutes with appr. 75 ml of Et20, and the insoluble white solid is filtered off and washed with Et20.
The filtrate contained most of the benzamide and 2-methylthio components. A solution of the Et20-insoluble solid (1.38 g) in MeOH is evaporated with appr. 25 g of silica gel. The powdered residue is layered evenly onto the top of a silica gel column, which is then eluted with CHC13/MeOH/HOAc (95:5:1) to give the 2-methylthio by-product and the desired 2-amino product (IVA). (IVA) is recrystallized by extraction into boiling isopropyl acetate in a Soxhlet apparatus. The white crystals are collected in three crops and dried in vacuo over P205 at 110 °C for WO 91/06548 PCT/US90l05756 ~r,,V>
~D'~
.J I IV.f.~i~i~
7h; yield 51.6%; mp >350 'C dec.; anal. calcd. for C17H22N40'0.21MeOH~0.22H20: C, 66.88; H, 7.59; N, 18.12.
Found: C, 66.86; H, 7.59; N, 18.12.

5 The compound of Example 104 is tested for enzyme inhibition activity. A purine nucleoside phosphorylase (PNP) enzyme assay is performed in which the PNP activity (IC50) for the compound is found, which is determined radiochemically by measuring the formation of [14C]-10 hypoxanthine from [14C]-inosine (see Biomedicine, 33, 39 (1980)) using calf spleen as the enzyme source. At 1 mM
phosphate the IC50 is 0.090 uM, and at 50 mM phosphate the IC50 is 2.5 ~M.

15 The following compounds of the present invention are r prepared that are 2-amino-7-(R)-3H,5H-pyrrolo[3,2-d]-pyrimidin-4-ones wherein R is -CH2-R1 in which the R1 group is a 2-adamantyl group as follows:
Example 106 R1 = 2-(1-methyl)-adamantyl 20 Example 107 R1 = 2-(1-chloro)-adamantyl Example 108 R1 = 2-(1-trifluoromethyl)-adamantyl Example 109 R1 = 2-(1-methoxy)-adamantyl Example 110 R1 = 2-(1-fluoro)-adamantyl The compounds are prepared following the procedures set 25 forth in Examples 98-104 using the appropriate 3-(2 adamantyl)-propio-nitriles as starting materials.

The following compounds of the present invention are prepared that are 2-amino-7-(R)-3H,5H-pyrrolo[3,2-d]
30 pyrimidin-4-ones wherein R is -CH2-R1 in which the R1 group is a 1-adamantyl group as follows:
Example 111 R1 = 1-(2-methyl)-adamantyl Example 112 R1 = 1-(2-chloro)-adamantyl Example 113 R1 = 1-(2-trifluoromethyl)-adamantyl WO 91!06548 PCT/US90/05756 . . ~d?! °a'~
:"r.....;.,.

Example 114 R1 = 1-(2-methoxy)-adamantyl Example 115 R1 = 1-(2-fluoro)-adamantyl Example 116 R1 = 1-adamantyl The compounds are prepared following the procedures set forth in Examples 98-104 using the appropriate 3-(1 adamantyl)-propio-nitrites as starting materials.

A pharmaceutical composition for intraperitoneal injection is prepared for testing the compound (IVA). An intraperitoneal injection solution containing the compound (IVA) is dissolved in an aqueous carrier that contains ten percent DMSO.

The compound (IVA) is intraperitoneally injected into Lewis Rats via the test composition of Example 117 to .
provide 30 mg of the compound (IVA), with an injection given twice per day. Controls are used, which receive only the vehicle. At specific times after administration, the animals are sacrificed and plasma samples are prepared.
The plasma is extracted with cold 0.5 N HC104 and neutralized with solid NH4HC03. After removal of perchlorate salts, the extract is subjected to HPLC on a reversed phase column (Spherisorb ODSI). A significant increase in plasma inosine is observed in the plasma taken from animals receiving the compound (IVA).

Compounds prepared as in Examples 106-116 are each made into a pharmaceutical formulation in accordance with the preparation of Example 117 and the resultant injectable solutions are tested in accordance with the procedure of Example 118. A significant increase in plasma inosine is observed in the plasma taken from animals receiving the compounds of the present invention.

WO 91/06548 PCI'/US90/05756 3-Cyclopentylpropionitrile is prepared in this example.
3-Cyclopentylpropionyl chloride (57.7 g, 0.36 mole) is added dropwise to a large excess of concentrated ammonium hydroxide (400 ml) cooled in an ice/salt bath. The heavy suspension of white solid is stirred overnight, collected by filtration, washed with cold water, and recrystallized fram about 2 liters of boiling water. The lustrous white plates of the amide are dried in vacuo over P205; yield 31.6 g (62.3%); mp 122 °C.
With protection from atmospheric moisture, a solution of the amide (23.5 g, 0.166 mole) in POC13 (150 ml) is heated at 120 °C for 1 h. The oil bath is cooled to about 70 °C, and excess POC13 is distilled off under vacuum, and the cooled residue is poured onto ice (about 300 g). The mixture is neutralized by cautious addition of solid Na2C03 and extracted with several portions of Et2O. The dried (Na2S04) extract is evaporated to give a clear, pale yellow oil which is distilled in vacuo to give the desired nitrile; yield, 16.86 g (82%) by 88.0 - 88.5 °C/8.7 mm).
MS (EI): m/z 122 (M-H)+; IR (cap. film), 2245cm-1(CN); 1H
NMR, d 1.67 (q, 2, -CH2CH2CN), 2.36 (t, 2, -CH2CN), complex multiplets centered about 1.11, 1.63, 1.86 (cyclopentyl protons).

3-Cyclopentylpropionitrile of the previous example is further treated in the synthesis of the present invention.
Under an atmosphere of dry N2, a mixture of 3-cyclopentyl-propionitrile (14.8 g, 0.12 mole), sodium hydride (5.8, 0.24 mole), and anhydrous tetrahydrofuran (30o ml) is heated at 52 °C in a water bath for 15 min., and a solution of ethyl formate (13.3 g, 0.18 mole) in THF (100 ml) is added over a period of 45 min. After two houxs at 50 - 55 °C, a second portion of NaH (1.9 g) and HC02Et (5.0 ml) are WO 91/06548 PCf/US90/05756 ~~ ~f.M./:.., iG~_.h-(n~r~~l ~ ;,~
added, followed in 30 min. by a third portion of HC02Et (unreacted nitrile is inert in the next step and can be recovered at the first purification step). The thick paste is stirred overnight and allowed to cool to room temperature. Volatile matter is evaporated under reduced pressure, and the residual pale yellow crust is dissolved in the minimum volume of cold water (about 150 ml) at 0'C.
The solution is adjusted to pH 6.0 by addition of 6N HCl and extracted with CHC13 (3 x 100 ml). The extract is washed with H20, dried over Na2S04; and evaporated in vacuo to a thick amber oil. This crude product (15.6 g) is used in the next reaction without further purification.

Glycine methyl ester hydrochloride (19.31 g, 0.154 mole) and anhydrous sodium acetate (12.61 g, 0.154 mole) are added to a solution of the crude formyl compound of the previous example (15.6 g) in MeOH/H20 (4:1, 500 ml). After 24 hours, the MeOH is evaporated in vacuo, and the mixture of water and oil is extracted with CHC13. The CHC13 layer 2o is dried (Na2S04) and evaporated to give an amber oil which is applied to a silica gel column. Elution with CHCl3 gave two major bands: (1) 3-cyclopentylpropionitrile (8.22 g, 66.: mmole or 55.6% of the nitrile used as starting material in the previous step), and (2) the desired enamine (3.45 g; 29.1% based on theoretical yield corrected for amount of nitrite present in starting material; MS (FAB):
223 (M + H)+) .

Under a nitrogen atmosphere, ethyl chloroformate (2.53 g, 23.3 mmole) is added dropwise to a solution of the enamine of the previous example (3.45 g, 15.5 mmole) and DBN (5.78 g, 46.6 mmole) in dry CH2C12 (50 ml) with external cooling in an ice bath. After stirring at 0 °C for one hour, the solution is allowed to a stand at room temperature ~~~.r-w,r~,..~ .~
tGo.~~~ r:w.,.::.::~

overnight. After checking progress by TLC, additional C1C02Et (0.5 ml) and DBN (3.0 ml) are added to complete the conversion, and the solution is allowed to stand for 24 hours. Volatile matter is evaporated in vacuo, the viscous residue purified on a short silica gel column (whose main purpose is to remove the less-mobile DBN) to give an N-blocked pyrrole (4.50 g; 98%), which is used for the next step without further purification.

l0 To a solution of the N-blocked pyrrole of the previous example (4.50 g, 15.3 mmole) in MeOH (100 ml) is added solid Na2C03 (1.62 g, 15.3 mmole), and the reaction mixture is stirred at room temperature for 48 hr with separation of the resultant deblocked pyrrole. The mixture is evaporated to dryness, and the residue is triturated thoroughly with H2o (50 ml) to dissolve the inorganics and extracted with CHC13 (3 x 100 ml). The extract is dried (Na2S04) and evaporated to give a viscous gum that crystallized upon drying in vacuo; yield 2.97 g (87.4%) of material suitable for use as an intermediate without further purification.
More extensive purification can, however, be effected by using either column chromatography employing silica gel/CHC13 or recrystallization from toluene/cyclohexane (1:3) .

Benzoyl isothiocyanate (2.62 g, 16.03 mmole) is added dropwise to a solution of the unblocked pyrrole of Example 134 (2.97 g, 13.36 mmole) in dry CH2C12 (100 ml). After 1 h at room temperature, solution is evaporated, and the gummy residue is dissolved in Et20 (100 ml) with almost immediate separation of crystalline solid; yield 1.75 g.
The Et20 filtrate is heated to bailing and diluted with an equal volume of warm cyclohexane. On cooling slowly the solution gave an additional 1.58 g of thioureido product;

WO 91/06548 PCf/US90/05756 ~.;~-,~~"~
~~... r;.",.,:_;~~
total yield 3.33 g (64.6%). A small amount of the thioureido product is recrystallized from warm Et20/cyclohexane (15 ml each); mp 123 - 125 °C. MS (FAB):
386 (M + H)+. Anal. Calcd, for C20H23N303S~0.45C6H12: C.
5 64.40; H, 6.76; N, 9.93. Found: C, 64.51. H, 7.10 N, 9.93.
EXAMPLE lab Methyl iodide (2.60 g, 18.32 mmole) is added to a solution of the thioureido product of Example 135 (3.21 g, 10 8.33 mmole) and DBN (1.24 g, 9.99 mmole) in dry CH2C12 (80 ml) at 0 °C. The solution is stirred at 0 °C for 15 min., at ambient temperature for 1 h, and then evaporated in vacuo. A solution of the residue in CHC13 is chromatographed on a silica gel column with CHC13 as eluent 15 to give homogeneous fractions of the methylthio intermediate compound; yield, 2.46 g (74%).

A solution of the methylthio compound of Example 136 (2.07 g, 5.18 mmole) in 150 ml --of MeOH that has been 20 saturated With NH3 at 0 'C is 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 the compound (IVC), benzamide and a by-product that is a 2-methy?thio derivative, as opposed to the 2-25 amino compound (IVC). The mixture is stirred vigorously for several minutes with appr. 75 ml of Et2o, and the insoluble white solid is filtered off and washed with Et20.
The filtrate contained most of the benzamide and 2-methylthio components. A solution of the Et20-insoluble 30 solid (1.13 g) in MeOH is evaporated with appr. 10 g of silica gel. The powdered residue is layered evenly onto the top of a silica gel column, which is then eluted with CHC13/MePH/HOAc (95:5:1) to give the 2-methylthio by-product (252 mg; MS (FAB) : 264 (M + H)+) and the desired ..>

2-amino product (IVC) (679 mg, 56.4%). (IVC) is recrystallized by extraction into boiling isopropyl acetate in a Soxhlet apparatus. The white crystals are collected in three crops and dried in vacuo over P205 at 110 °C for 7h; yield 5.40 mg (44.9%) ; mp 324 - 326 ° C dec. ; MS (FAB) 233 ( M + H)+: anal. calcd. for C12H16N40~ C, 62.05; H, 6.94; N, 24.12. Found: C, 62.04: H, ?.11; N, 24.48.

The compound of Example 137 is tested for enzyme°
inhibition activity in accordance with the procedure of Example 9. At 1 mM phosphate the IC50 is 0.029 ~,M, and at 50 mM phosphate the IC50 is 1.8 ~,M.

The following compounds of the present invention are prepared that are 2-amino-7-(R)-31_i,SH_-pyrrolo[3,2-d]-pyrimidin-4-ones wherein R is -CH2-R1 in which the R1 group is as follows:
Example 139 R1 = 3-methylcyclopentyl Example 140 R1 = 2-chlorocyclopentyl Example 141 R1 = 3-triflouromethylcyclopentyl Example 142 R1 = 3-methoxycyclopentyl The compounds are prepared following the procedures set forth in Examples 130-137 using the appropriate 3 (substituted cyclopentyl)-propionitriles as starting materials.

A pharmaceutical composition for intraperitoneal injection is prepared for testing the compound (IVC). An intraperitoneal injection solution is prepared containing the compound (IVC) is dissolved in an aqueous carrier that contains ten percent DMSO.

f %~M.L..i~RY

Using the procedure of Example 112, the compound (IVC) is intraperitoneally injected into Lewis Rats via the test composition of Example 143 and the results compared with controls. A significant increase in plasma inosine is observed in the plasma taken from animals receiving the compound (IVC).

3-Cyclohexylpropionitrile is prepared in this example. A
solution of cyclohexanepropionic acid (50 g; 0.32 mole) and thionyl chloride (152 g; 1.28 mole) in 100 ml benzene is allowed to stand overnight and is then evaporated to an oily residue. The residue is added in portions to 28%
aqueous ammonia (270 ml) at 25°C and the mixture stirred for about two hours. The resulting product is collected by filtration, washed with cold water, and recrystallized from about 2 liters of boiling water. The lustrous white plates of the amide are dried in vacuo over P205; yield 45.5 g.
With protection from atmospheric moisture, a solution of the amide (45.5 g, 0.293 mole) in SOC12 (200.3 g; 1.68 mole) refluxed for about six hours. The oil bath is cooled to about 70 °C, and excess SOC12 is distilled off under vacuum, and the cooled residue is poured onto ice (about 300 g). The mixture is neutralized by cautious addition of solid Na2C03 and extracted with several portions of Et20.
The dried (Na2S04) extract is evaporated to give a clear, pale yellow oil Which is distilled in vacuo to give the desired nitrile; yield 42.0 g.

3-Cyclohexylpropionitrile of Example 145 is further treated in the synthesis of the present invention. Under an atmosphere of dry N2, a mixture of 3-cyclohexyl-propionitrile (22.3 g, 0.16 mole), sodium hydride (5.38, 0.224 mole), and anhydrous tetrahydrofuran (120 ml) is heated at 52 °C in a water bath for 15 min., and a solution of ethyl formate (55.4 g, 0.75 mole) in THF (50 ml) is added over a period of 45 min. After two hours at 50 - 55 °C, a second portion of NaH (2.0 g) and HC02Et (5.0 ml) are added (unreacted nitrile is inert in the next step and ca be recovered at the first purification step), and the reaction mixture is stirred for about three days at 55'C
and then allowed to cool to roam temperature. Volatile matter is evaporated under reduced pressure, and the residual pale yellow crust is dissolved in the minimum volume of cold water (about 75 ml) at 0 °C. The solution is adjusted to pH 6.0 by addition of 6N HC1 and extracted with CHC13 ( 3 x 100 ml ) . The extract is washed with H20, dried over Na2S04, and evaporated in vacuo to a thick amber oil. This crude product is used in the next reaction without further purification.

Glycine methyl ester hydrochloride (30.60 g, 0.24 mole) and anhydrous sodium acetate (19.99 g, 0.24 mole) are added to a solution of the crude formyl compound of the previous example (25.22 g) in MeOH/H20 (4:1, 500 ml). After 24 hours, the MeOH is evaporated in vacuo, and the mixture of water and oil is extracted with CHC13. The CHC13 layer is dried (Na2S04) and evaporated to give an amber oil which is applied to a silica gel column. Elution with CHC13 gave two major bands: (1) 3-cyclohexylpropionitrile (used as starting material in the previous step), and (2) the desired enamine: yield 16 g.

Under a nitrogen atmosphere, ethyl chloroformate (1.38 g, 12.7 mmole) is added dropwise to a solution of the enamine of Example 147 (2.0 g, 8.46 mmole) and DBN (2.1 g, 16.9 morale) in dry CH2C12 (50 ml) with external cooling in an ice bath. After stirring at 0 °C for one hour, the WO 91/Ob~48 PGT/US90/0575b I~ ~"tI .'r ~;s~ f IM.i:v~'t11 solution is allowed to a stand at room temperature overnight. After checking progress by TLC, additional C1C02Et (0.5 ml) and DBN (1.5 ml) are added to complete the conversion, and the solution is allowed to stand for 24 hours. Volatile matter is evaporated in vacuo, the viscous residue purified on a short silica gel column (whose main purpose is to remove the less-mobile DBN) to give an N-blocked pyrrole, which is used for the next step without further purification.

To a solution of the N-blocked pyrrole of Example 148 (2.6 g, 8.43 mmole) in MeOH (100 ml) is added solid Na2C03 (2.23 g, 21.07 mmole), and the reaction mixture is stirred at room temperature for 48 hr with separation of the resultant deblocked pyrrole. The mixture is evaporated to dryness, and the residue is triturated thoroughly with H2o (50 ml) to dissolve inorganics and extracted with CHC13 (3 x 100 ml). The extract is dried (Na2S04) and evaporated to give a viscous gum, which is purified on a silica gel column using CHC13 as the eluent; yield 1.67 g (84%); m.p.
73-74'C.

Benzoyl isothiocyanate (0.74 g, 4.02 mmole) is added dropwise to a solution of the unblocked pyrrole of Example 149 (0.95 g) in dry CH2C12 (20 ml). After one hour at room temperature, the solution is evaporated, and the gummy residue is dissolved in Et20 (100 ml) with almost immediate separation of crystalline solid. The Et2o filtrate is heated to boiling and diluted with an equal volume of warm cyclohexane. On cooling slowly the solution gives additional thioureido product; total yield 1.41 g (88%):
m.p. 156-157°C.

~~,lM.~~ ~ W1 FF~~ldd01 J s'~e.~l..:~n.;J~

Methyl iodide (1.1 g, 7.6 mmole) is added to a solution of the thioureido product of Example 150 (0.96 g, 2.61 mmole) and 1,5-diazabicyclo[4.3.0]non-5-ene (0.38 g, 3.0 5 mmole) in dry CH2C12 (20 ml) at 0°C. The solution is stirred at 0°C for 15 min., at ambient temperature for 1 h, and then evaporated in vacuo. A solution of the residue in CHC13 is chromatographed on a silica gel column with CHC13 as eluent to give homogeneous fractions of the methylthio 10 intermediate compound; yield 0.92 g.

A solution of the methylthio compound of Example 151 (0.8 g, 1.93 mmole) in 50 ml of MeOH that has been -saturated with NH3 at 0 °C is heated at 90-95 °C for 24 15 hours in a glass-lined stainless steel bomb. The contents of the chilled bomb are evaporated in vacuo to give a mixture of the compound (IVD), benzamide and a by-product that is a 2-methylthio derivative, as opposed to the 2-amino compound (IVD). The mixture is stirred vigorously 20 for several minutes with appr. 75 ml of Et20, and the insoluble white solid is filtered off and washed with Et2o.
The filtrate contained most of the benzamide and 2-methylthio components. A solution of the Et2o-insoluble solid (0.390 g) in MeOH is evaporated with appr. 10 g of 25 silica gel. The 'powdered residue is layered evenly onto the top of a silica gel column, which is then eluted with CHC13/MeOH/HOAc (95:5:1) to give the 2-methylthio by-product and the desired 2-amino product (IVD). (IVD) is recrystallized by extractian into boiling isopropyl acetate 30 in a Soxhlet apparatus. The white crystals are collected in three crops and dried in vacuo over P205 at 110 °C for 7h; yield 49%, mp >300 °C; anal. calcd. for C13H18N40: C, 63.39, H, 7.36: N, 22.74. Found: C, 63.50; H, 7.74: N, 22.67.

WO 91/065x8 PCT/US90/05756 t.:N.a..' v The compound of Example 152 is tested for enzyme inhibition activity as in Example 105. At 1 mM phosphate the IC50 is 0.037 uM, and at 50 mM phosphate the IC50 is 2.2 uM.

The compound of Example 152 is tested to determine its effectiveness in potentiation of the toxicity of 2'-deoxyguanosine (d-Guo) (see D. A. Schewach et al., Cancer -ss. , 46, 519 (1986) , and J. C. Sircar et al. , Accents and ~tions, 21, 253 (1987)). CCRF-CEM cells are grown in RPMI-1640 medium. To a suspension cultures of these cells, d-Guo at a fixed concentration (5.62 ~,M) and the compound at varied concentrations are added and the number of cells are determined in a Coulter counter 24, 48, and 72 hours thereafter. From these data, the IC50 is calculated to be 2.0 ~,M as the concentration of the compound required to reduce the increase in cell number between 0 and 72 hours to 50% of that of control cultures.

The compound 2-amino-7-(3-methylcyclohexylmethyl)-3~i,5H_-pyrrolo[3,2-d]pyrimidin-~!-one is prepared. First, using the procedures set forth in Examples 146-152 above, but with 3-(3-methylbenzyl)-propionitrile as the starting material, the aryl derivative 2-amino-7-(3-methylbenzyl)-3H,5H-pyrrolo[3,2-d]-pyrimidin-4-one is made. A solution of the aryl derivative (0.2 g, 0.78 mmole) in trifluoroacetic acid (TFA) (20 ml) is hydrogenated with Pto2 at 60 lb/in2 for 24 h. Catalyst is filtered off through a Celite bed, and the filtrate is evaporated. The residue is triturated with methanol and left in the refrigerator overnight. The resulting crystallized trifluoroacetate salt precipitates from the solution and is collected by filtration. The TFA salt is suspended in 8 ml . f~,~ s ~A ~y~..j1 n.>

of H20, adjusted to pH8 by cone. NH40H and sonicated. The pure product is collected, washed with H20 and dried: yield 165 mg (81%); mp 282 °C. Anal: Calcd. for C14H20N40= . C, 64.60; H, 7.74; N, 21.52. Found: C, 64.24; H, 7.96; N, 21.51%.

The procedure described in Example 155 is repeated to prepare 2-amino-7-(3-trifluoromethylcyclohexyl-methyl)-3~,5~i-pyrrolo-(3,2-d]pyrimidin-4-one using 3-(3-trifluoromethylbenzyl)-propionitrile as the starting compound: yield 69%; mp 165 'C. Anal. caled. for C14H17N40F3'0.6H20: C, 51.72 H, 5.64; N, 17.23. Found:
C, 51.82; H, 5.71: N, 16.81%.

The compound prepared in Example 155 is tested for enzyme-inhibition activity as in Example 105. At 1 mM
phosphate the IC50 is 0.025 ACM, and at-50 mM phosphate the IC50 is 0Ø820 ~M.

The compound prepared in Example 156 is tested for enzyme-inhibition activity as in Example 105. At 1 mM
phosphate the IC50 is 0.020 ~M, and at 50 mM phosphate the IC50 is 0.740 ~M.

3-Cycloheptylpropionitrile is prepared in this example according to the procedure of Example 97 using a solution of 2-bromocycloheptane (25.57 g; 144.38 mmole); Bu3SnH
(50.42 g; 173.26 mmole), acrylonitrile (15.32 g; 288.77 mmole), and AIBN (1.13 g) in toluene (300 ml). Yield is 16 g; mp oil.

3-Cycloheptylpropionitrile of Example 159 is further treated in the synthesis of the present invention. Under an atmosphere of dry N2, a mixture of 3-cycloheptyl-., ..d J'..:WI
n~F,f mp~ s~~,~

propionitrile (8.5 g, 56.19 mmole), sodium hydride (2.6 g, 112.39 mmole), and anhydrous tetrahydrofuran (100 ml) is heated at 52 °C in a water bath for 15 min., and a solution of ethyl formate (20.81 g, 280.99 mmole) in THF (100 ml) is added over a period of 45 min. After two hours at 50-55'C, a second portion of NaH (1.35 g) and HC02Et (10.4 g) are added, followed in 30 min. by a third portion of HC02Et (unreacted nitrile is inert in the next step and can be recovered at the first purification step). The thick paste is stirred overnight and allowed to cool to room temperature. Volatile matter is evaporated under reduced pressure, and the residual pale yellow crust is dissolved in the minimum volume of cold water (about 150 ml) at 0'C.
The solution is adjusted to pH 6.0 by addition of 6N HC1 and extracted with CHC13 (3 x 100 ml). The extract is Washed with H20, dried over Na2S04, and evaporated in vacuo to a thick amber oil. This crude product is used in the next reaction without further purification.

Glycine methyl ester hydrochloride (9.35 g, 74.47 mmole) and anhydrous sodium acetate (6.10 g, 74.47 mmole) are added to a solution of the crude formyl compound of Example 160 (8.9 gt 49.65 mmole) in MeOH/H20 (4:1, 250 ml). After 24 hours, the MeOH is evaporated in vacuo, and the mixture of water and oil is extracted with CHC13. The CHC13 layer is dried. (Na2S04) and evaporated to give an amber oil which is applied to a silica gel column. Elution with CHC13 gave two major bands: (1) 3-cycloheptylpropionitrile (used as starting material in the previous step), and (2) the desired enamine, which is recrystallized from a CHC13/Et20 mixture; yield 6.18 g; m.p. 57-58°C.

Under a nitrogen atmosphere, ethyl chloroformate (4.01 g, 37.03 mmole) is added dropwise to a solution of the enamine ~~ ;t ~, ~~~:.a ..

of Example 161 (6.18 g, 24.69 mmole) and DBN (9.19 g, 74.04 mmole) in dry CH2C12 (100 ml) with external cooling in an ice bath. After stirring at 0 °C for one hour, the solution is allowed to a stand at room temperature overnight. After checking progress by TLC, additional C1C02Et (0.5 ml) and DBN (3.0 ml) are added to complete the conversion, and the solution is allowed to stand for 24 hours. Volatile matter is evaporated in vacuo, the viscous residue purified on a short silica gel column (whose main purpose is to remove the less-mobile DBN) to give an N-blocked pyrrole, which is used for the next step without further purification.

To a solution of the N-blocked pyrrole of Example 162 (7.8 g, 24.19 mmole) in MeOH (100 ml) is added solid Na2C03 (6.41 g, 60.48 mmole), and the reaction mixture is stirred at room temperature for 48 hr with separation of the resultant deblocked pyrrole. The mixture is evaporated to dryness, and the residue is triturated thoroughly with H2o (50 ml) to dissolve inorganics and extracted with CHC13 (3 x 100 ml). The extract is dried (Na2SO4) and evaporated to give a viscous gum, Which was purified by column chromatography employing silica gel/CHC13; yield 4 g; m.p.
88-X39 ° C.

Benzoyl isothiocyanate (1.5 g, 8.96 mmole) is added dropwise to a solution of the unblocked pyrrole of Example 163 (1.99 g, 7.95 mmole) in dry CH2C12 (50 ml). After 1 h at room temperature, solution is evaporated, and the gummy residue is dissolved in Et2o (l00 ml) with almost immediate separation of the crystalline solid. The Et2o filtrate is heated to boiling and diluted with an equal volume of warm cyclohexane. On cooling slowly the solution gives additional thioureido product; yield 2.89 g (88%); m.p.
158-159'C.

Methyl iodide (1.7 g, 11.96 mmole) is added to a salution 5 of the thioureido product of Example 164 (1.7 g, 4.1 mmole) and DBN (0.56 g, 4.52 mmole) in dry CH2C12 (80 ml) at 0 'C.
The solution is stirred at 0'C for 15 min., at ambient temperature for 1 h, and then evaporated in vacuo. A
solution of the residue in CHC13 is chromatographed on a 10 silica gel column with CHC13 as eluent to give homogeneous fractions of the methylthio intermediate compound.

A solution of the methylthio compound of Example 165 (1.72 g, 4.02 mmole) in 50 ml of MeOH that has been '!5 saturated with NH3 at 0 'C is 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 the compound (IVE), benzamide and a by-product that is a 2-methylthio derivative, as opposed to the 2-20 amino compound (IVE). The mixture is stirred vigorously for several minutes, with appr. 75 ml of Et20, and the insoluble white solid is filtered off and Washed With Et2o.
The filtrate contained most of the benzamide and 2-methylthio components. A solution of the Et20-insoluble 25 solid (0.850 g) in MeOH is evaporated with appr. 1o g of silica gel. The powdered residue is layered evenly onto the top of a silica gel column, which is then eluted with CHC13/MeOH/HOAc (95:5:1) to give the 2-methylthio by-product and the desired 2-amino product (IVE). (IVE) is.
30 recrystallized by extraction into boiling isopropyl acetate in a Soxhlet apparatus. The white crystals are collected in three crops and dried in vacuo over P205 at 110 °C far 7h: yield 54%, mp >300 °C dec.: anal. calcd. for C14H2pN40:

,..~-,,, ~ '° ;3 p ,.,, a._:.y ..

C, 64.60; H, 7.74; N, 21.52. Found: C, 64.78: H, 8.01; N, 21.61.

The compound of Example 166 is tested for enzyme inhibition activity as in Example 105. At 1 mM phosphate the IC50 is 0.030 ACM, and at 50 mM phosphate the IC50 is 0.840 uM.

Using the procedure of Example 97, 3-(1-norbornanyl)-propionitrile is made from 1-bromonorbornane, and 3-(2-norbornanyl)-propionitrile (mixture of 2-exo and 2-endo) is made from 2-bromonorbornane. Following Examples 98-104, the propionitriles are converted to the compounds (IVF), IV(G), and (IVH).

Using the procedure 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 made respectively from 1-bromo-bicyclo[3.2.1]octane, 2-bromo-bicyclo[3.2.1]octane, 3-bromo-bicyclo[3.2.1]octane, and 8-bromo-bicyclo[3.2.1]octane. Following Examples 98-104, the propionitriles are converted to the compound (IVL) and the related, 2-bicyclo[3.2.1]octanyl, 3-bicyclo[3.2.1]octanyl, and 8-bicyclo[3.2.1]octanyl derivatives.

Using a modification of the procedure disclosed 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 trimethylsulfoxonium iodide, giving an intermediate epoxide, which is then converted to the aldehyde by treatment with boron trifluoride etherate.
Following the procedure of Netherlands Pat. 6,610,204, the .~ ~-., r .'rA ~i ';Z "~
,C,s ~. ; ! /.., .i...'-~ a.

aldehyde is condensed with cyanoacetic acid by refluxing in a pyridene/toluene solution with a catalytic quantity of ammonium acetate for 48-72 hours to give the corresponding acrylonitrile. The acrylonitrile is then hydrogenated using a palladium-on-carbon catalyst in methanol as taught in Profitt, et al, J. Orq. Chem., 40, 127 (1975) to give 3 (6-bicyclo-[3.2.1)octanyl)-propionitrile. Following Examples 98-104, the propionitrile is converted to the 6 bicyclo[3.2.1]octanyl derivative related to the compound ( IVL) .

Using the procedure of Example 97, 3-(1-bicyclo[3.3:1]
nonanyl)-propionitrile and 3-(3-bicyclo-[3.3.1]nonanyl)-propionitrile are respectively made from 1-bromo-bicyclo[3.3.1]nonane and 3-bromo-bicyclo[3.3.1]nonane.
Following Examples 98-104, the propionitriles are converted to the compound (IVM) and the related 3-bicyclo[3.3.1]-nonanyl derivative.

Following the procedure of Example 171, bicyclo[3.3.1]
nonane-9-one is reacted to form the corresponding aldehyde, from which is made the corresponding 3-substituted propionitrile, which is then converted into the 9 bicyclo[3.3.1]nonanyl derivative related to the compound (IVM).

Following the procedure of Example 170, 2-bicyclo[3.3.1]-nonanecarboxaldehyde is reacted to form the corresponding 3-substituted propionitrile, which is then converted into the 2-bicyclo[3.3.1]nonanyl derivative related to the compound (IVM).

Using the procedure of Example 97, 3-(1-noradamantyl)-propionitrile is made from 1-bromonoradamantane, and 3-(2-.,..;,~ ~-, ~ , ~'~' f : ~ :.
,, ..m.,.~. 58 noradamantyl)-propionitrile is made from 2-bromonoradamantane. Following Examples 98-104, the propionitrile is converted to the final compound (IVN).

Following the procedure of Example 170, 3-noradamantane-carboxaldehyde is reacted to form the corresponding 3-substituted propionitrile, which is then converted into the 3-noradamantyl derivative related to the compound (IVN).

l0 Following the procedure of Example 170, noradamantane-7-one is reacted to form the corresponding aldehyde, from which is made the corresponding 3-substituted propionitrile, which is then converted into the 7-noradamantyl derivative related to the compound (IVN).

Using the procedure of Example 9?, 3-(1-bicyclo[2.2.2]-octanyl)propionitrile is made from 1-bromobicyclo(2.2.2]-octane and 3-(2-bicyclo[2.2.2]octanyl)propionitrile is made from 2-bromobicyclo[2.2.2]octane. Following Examples 98-104, the propionitriles are converted to the compound (IVK) and the related 2-bicyclo[2.2.2]octanyl derivative.

Using the procedure of Example 97, 3-(1-norbornenyl) propionitrile is made from 1-bromonorbornene. Following Examples 98-104, the propionitrile is converted to the compound (IVI).

Following the procedure of Example 170, 5-norborene-2 .carboxaldehyde (a mixture of 2-endo and .2-exo) is reacted to form the corresponding 3-substituted propionitrile, which is then converted into the compound (IVJ).

' '1'~'~~~ r' 59 a~o~~' ~., .3' ~M.i...~'..'~

N
CI
~ C~~ N

The above intermediate compound is prepared in this Example by the modification of the procedure of Schiemenz, G. P.; Engelhard, H. CChem. 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 reflux for l6 h in a flask fitted with Dean-Stark trap and condenser. The solvents are evaporated in vacuo, residue is extracted with CHC13, which is washed With H2O, dried (Na2SO4), and evaporated to give the crude product, which is purified by silica gel column chromatography using hexane-EtOAc mixture as the eluent. Yield 23.69 g (73%)t mp 90-91 °C.

N
C1 ~ CHO
(~-J) Cl CN

The above intermediate compound is prepared in this Example. To a stirred mixture of NaH (1.56 g, 65.05 mmol) and ethyl formate (14.78 g, 199.51 mmol) in THF (100 ml) is added substituted pentanedinitrile of Example 180 (10.17 g, 37.17 mmol) at room temperature under a nitrogen WO 91/0654$ PCT/US90/05756 ~~,~~.aa.~
r.
atmosphere, and the resulting reaction mixture is stirred for 24 h. Volatile matter is evaporated in vacuo at room temperature. Water (50 ml) is added to the residue at 0-5 °C, and the solution is adjusted to pH 5-6 by 20% conc. HC1 5 (v v). The heavy oil is extracted into ethyl acetate, washed with H2o (1 x 100 ml) and dried (MgSO4). The ethyl acetate layer is evaporated to give a red-brown oil (11.0 g) that is used in the next step without further purification.

'~~~OMe C1 ~h.
15 C~ CN
The above intermediate compound is prepared in this Example. Glycine methyl ester hydrochloride (8.17 g, 65.06 mmol) and sodium acetate (5.33 g, 65.06 mmol) are added to 20 a solution of the crude formyl compound of Example 181 (11.0 g) in a mixture of MeOH (80 ml) and H20 (20 ml), and the resulting solution is stirred at room temperature for 22 h. After evaporation of solvent at room temperature, the residue is extracted with ethyl acetate. The washed 25 (H20) and dried (MgS04) organic layer is evaporated to give an oil. Flash column chromatography (silica gel) using CHC13 as eluent gave the pure desired enamine as a mixture of cis-trans isamers which is recrystallized from MeOH, yield 10.41 g (75%), mp 142-143 °C.

WO 91/06548 PCt'/US90/05756 ~-~,r . Ta.,~ '~.~
t t , .,. _: ° .

CI Et~O ( 0 N
~OMe C1 \ / NHS
~cy The above intermediate compound is prepared in this Example. A solution of enamine of Example 182 (10.0 g, 26.84 mmol) in dry CH2C12 (100 ml) is cooled to 0 °C and treated with 1,5-diazabicyclo[4.3.0]non-5-ene (10.53 g, 84.79 mmol) under a nitrogen atmosphere followed by ethyl chloroformate (6.90 g, 63.57 mmol). The solution is stirred at 0 °C for 1 h and then at room temperature for 48 h. Volatiles are evaporated in vacuo to give a viscous dark gum which is purified by flash column chromatography over silica gel using CHC13 as the eluent. All the fractions containing the desired N-protected pyrrole are pooled and evaporated to give a foamy light pale yellow material which is stirred in MeOH (100 ml) to give the crystalline material Which is recrystallized from CHC13-MeOH, yield 8.92 g (74.7%), mp 160-161 °C.

N
1 ~ 'OMe C1 ~~ 'NHZ

The above intermediate compound is prepared in this 3o Example. A suspension of N-protected pyrrole of Example 183 (8.6 g, 19.34 mmol) in MeOH (300 ml) is treated with Na2C03 (5.12 g, 48.34 mmol) and the reaction mixture is stirred at room temperature for 17 h with separation of the deblocked pyrrole during the first hour. Solid sodium WO 91/06548 PC1'/US90/05756 ~~..'',..;..-~,~ ~r f LJ .L..~'~D

carbonate is removed by filtration and washed well with MeOH. The filtrate is reduced to a volume of 25 ml and kept in a refrigerator for 1 h to give 5.23 g of crystalline product. Further concentration of the mother liquor gave an additional 0.14 g of pure product; total yield 6.45 g (89.5 %), mp 7.78-181 °C.

~OMe C1 ~NH-C-NHCOPh The above intermediate compound is prepared in this Example. To a suspension of pyrrole of Example 184 (5.83 g, 15.64 mmol) in dichloromethane (100 ml) is added benzoylisothioeyanate (2.88 g, 17.64 mmol) at room temperature under nitrogen. The reaction mixture is stirred for 30 min with the separation of the desired thioureido compound. Additional benzoyl isothiocyanate (0.5 ml) is added to it and again stirred for 30 min. The solvent is evaporated to dryness, and the light yellow residue is triturated with methanol. The white crystalline material is isolated by filtration and recrystallized from a chloroform-ether mixture to give the required thioureido compound as an analytically pure sample, yield 7.71 g (92%), mp 210-211 °C.

~ Me C1 ~N=j~NHCOPh CI N SMe ~fi ' f ;~'~ .'?'.'.3 '.,r.m..: a The above intermediate compound is prepared in this Example. A solution of thioureido compound 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 CH2C12 (200 ml) is cooled to 0 °C and treated with methyl iodide (5.20 g, 36.65 mmol).
The reaction mixture is stirred at 0 °C for l0 min and then for 1 h at room temperature. The solvent is evaporated at room temperature, and the residue is extracted with CHC13, washed with H20 (2 x 50 ml), dried (Na2S04) and evaporated to give a glassy foam (6.95 g) which is used in the next step without purification.

E
Hz,I hte Cl H

A
The above compounds A and B are prepared in this Example.
The compound A is a compound of the present invention and the compound B is an intermediate. A solution of the methylthio intermediate of Example 186 (6.90 g, 12.54 mmol) in MeOH (200 ml) is saturated at 0 °C with ammonia and heated at 100 °C for 20 h in a glass-lined stainless steel bomb. The reaction mixture is brought to room temperature and the solvent is evaporated at room temperature.
Purification of the crude mixture by flash column chromatography over silica gel using CHC13 as eluent gave 8B (1.1 g, 21°s), mp 290-291 °C then CHC13-MeOH (95:5) gave pure 8A (2.76 g, 57.5%), mp 284-285 °C.

.~z~'r~~
~-; s~;,.,.r.'~''"

The compound of the present invention of Example 187 is tested for enzyme inhibition activity. A purine nucleoside phosphorylase (PNF) enzyme assay is performed in which the PNP activity (IC50) for the compound (8A) is found, which is determined radiochemically by measuring the formation of (14C]-hypoxanthine from (14C]-inosine (see Biomedicine, 1980, 33, 39) using calf spleen as the enzyme source. At 1 mM phosphate the IC50 is 0.64 uM and at 50 mM phosphate the IC50 is 10 ~,M.

H

Following the procedure set forth in Examples 180-187, 3-(3-chlorophenyl)-3-(2-amino-4-oxo-3H,5H-pyrrolo(3,2 ~]pyrimidin-7-yl)propanenitrile is prepared using 3-(3 chlorophenyl)-pentanedinitrile as the starting material, yield 54.5%, mp 157-158 °C.

Following the procedure set forth in Examples 180-187, the following compounds are also prepared (1-9).

H
H
H~ N~N I ~
Ar CN

~;'! ~M ~_.'-. W
3-Aryl-3-(2-amino-4-oxo-3H,5H-pyrrolo[3,2-d]
pyrimidin-7-yl)propanenitrile Where Ar is each of the following: (1) phenyl, 2,3 dichlorophenyl, 3-methylphenyl, and 3-methoxyphenyl, (2) 5 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-), and (9) pyrazolyl.

10 Following the procedure set forth in Examples 180-187, the following compounds 10-14 and 21 are prepared starting from the appropriately substituted pentanedinitrile.
Compounds 15-20, and 22 are prepared from the corresponding unsaturated Ar analogues in Example 190. In this 15 procedure, the nitrile group of the unsaturated analogue is first converted to an amide group by acid- or base-catalyzed hydrolysis, then the unsaturated Ar group is converted to the saturated R2 group by known catalytic hydrogenation, followed by reconverting the amide back to 20 the nitrile by known dehydration procedures.

H
H ( ' H2N ~ /
R N
3-(Substituted)-3-(2-amino-4-oxo-3H,5H-pyrrolo(3,2-d]-pyrimidin-7-yl)propanenitrile Where R2 is each of: 10) 1-adamantyl, 11) 2-adamantyl, 12) cyclohexyl, 13) cycloheptyl, 14) cyclopentyl, 15) tetrahydrofuranyl, 16) tetrahydrothienyl, 17) tetrahydropyranyl, 18) pyrazolidinyl, 19) thiazolidinyl, PCe~r~~A a f i..a.:,..

20) piperazinyl, 21) morpholinyl, and 22) hexahydropyridazinyl.

N
The above compound, 3-(2-amino-4-oxo-3H,5H_-pyrrolo[3,2-d_]pyrimidin-7-yl)-3-phenylpropanenitrile, is prepared in this Example. A solution of the compound A obtained in Example 187 (2.0 g, 5.22 mmol) in warm ethanol (250 ml) and dimethylformamide (DMF) (150 ml) is hydrogenated over 30%
Pd/C catalyst (1.0 g) in the presence of triethylamine (2.64 g, 5.0 equivalent) at atmospheric pressure. After 5 h, the reaction is complete, and the catalyst is filtered off under N2 pressure. The solid obtained by evaporation of the filtrate is triturated and sonicated With H20 and dried, yield 1.28 g (88%), mp 168-170 °C.

The compound prepared in Example 192 is tested for enzyme inhibition activity as in Example 188. At 1 mM phosphate the IC50 is 0.023 ACM and at 50 mM phosphate the IC50 is 4.7 ~tM .

a ~ Y~TT~ ~~
~~' ~:.,,.i...~ m.

The above compound, 3-(2-amino-4-oxo-3_H,5H-pyrrolo[3,2-_d]pyrimidin-7-yl)-3-phenylpropanoic acid, is prepared in this example. A solution of the compound obtained in Example 192 (0.200 g, 0.72 mmol) in 6N HC1 (3.0 ml) is heated at reflux for 18 h. The solvent is evaporated in vacuo and the residue is triturated with H2o (6 ml), adjusted to pH 10 by conc. ammonium hydroxide. Insoluble material is collected by filtration and the filtrate is readjusted to pH 6.8. White material which is precipitated out is collected, washed with water and dried, yield 0.19 g (89%), mp 290 °C dec.

The compound prepared in Example 194 is tested for enzyme inhibition activity as in Example 188. At 1 mM phosphate the IC50 is 0.012 ACM and at 50 mM phosphate the IC50 is 0.19 ~M.

Hz The above compound, 3-(Z-amino-4-oxo-3H,5H-pyrrolo[3,2-d_]pyrimidin-7-yl)-3-phenylpropanamide, is prepared in this example. A solution of the compound obtained in Example 192 (0.200 g, 0.72 mmol) in conc. H2S04 (0.5 m1) is stirred at room temperature for 20 h and then poured onto crushed ice (5.0 g) and adjusted to pH 6.8 by conc. NH40H. The precipitated solid is collected, washed with H2o and dried, yield 0.180 g, mp 199-201 °C dec.

The compound prepared in Example 196 is tested for enzyme inhibition activity as in Example 188. At 1 mM phosphate the IC50 is 0.20 ACM and at 50 mM phosphate the IC50 is 6.6 ~tM.

a The above compound, 3-(2-amino-4-oxo-3H,5H-pyrrolo[3,2-d_]pyrimidin-7-yl)-3-phenylpropanoic acid, methyl ester, is prepared in this example. Thionyl chloride (0..2 g, 0.17 mmol ) is added to stirred methanol ( 4 . 0 ml ) at 0 ° C. The compound obtained in Example 194 (0.2 g, 0.67 mmol) is added and the mixture is stirred at room temperature for 18 h. The solvent is removed on a water aspirator (30°C) and vacuum pump (lyophilize) to give a semisolid mass which is purified on a silica gel column using CHC13-MeOH as the eluent, yield 0.1 g.

The compound prepared in Example 198 is tested for enzyme inhibition activity. Significant activity (IC50) is found.

H., a WO 91/06548 PCT/US90l05756 n~YH'~~ ~.
.~~, ~ f i~M.J...: 9 3-(2-Amino-4-oxo-3H,5H-pyrrolo[3,2-d]pyrimidin-7-yl)-3-cyclohexylpropanoic acid is prepared in this example. A
solution of the compound obtained in Example 194 (83 mg, 0.28 mmol) in trifluoroacetic acid (TFA) (15 ml) is hydrogenated with Pt02 (83 mg) at 60 lb/in2 for 24 h. The catalyst is filtered off through a Celite bed, and the filtrate is evaporated at 25 °C. The residue is suspended in H20 (8 mI), and adjusted to pH 8.5 by cone. NH40H and filtered through a Whatman filter paper to remove brown colored impurities. The colorless filtrate is adjusted to pH 6.8, and the precipitated compound is filtered, washed with H20, and dried, yield 65 mg (77%), mp >300 °C.

The compound prepared in Example 200 is tested for enzyme inhibition activity as in Example 188. At 1 mM phosphate the IC50 is 0.097 ACM and at 50 mM phosphate the IC50 is 1.0 ~1M .

A compound of the present invention is prepared wherein X
is PO(OH)2. The nitrile group of the compound of Example 192 is converted to the corresponding amide by treatment with sulfuric acid. Using a Hoffman degradation reaction, the amide is converted to the corresponding amine, which is then converted to the corresponding pyridinium salt using a pyrillium salt. Conversion of the salt to the corresponding halide is accomplished using sodium bromide, which is then converted to the phosphonic ester using triethyl phosphate. Hydrolysis of the ester using trimethylsilylbromide yields the corresponding phosphonic acid wherein "n" is 1 and "m" is 0.

This Example makes a compound of the present invention by stepping up the number of carbon atoms from "m" is 0 to "m"
is 1. The nitrile group of the compound of Example 192 is .;

,~.~~-.r~ '.;.'.~
red~et~~f'°~'~o the corresponding aldehyde, which is then converted to the corresponding alcohol. Using phosphorous tribromide the alcohol is converted to the corresponding alkyl bromide, which is then converted to the nitrile 5 compound of the present invention Wherein m is 1 using potassium cyanide.

In this example a compound of the present invention is prepared wherein "p'~ is 1 and '~Y" is oxygen. The alcohol 10 prepared as an intermediate in the previous example is converted to the corresponding diethyl phosphonomethyl ether using diethylchloromethyl phosphonate. Removal of the ethyl groups of the ester is accomplished using trimethylsilylbromide to give the phosphonic acid.

In this example a compound of the present invention is made wherein "Y" is NH and "X" is S02NH2. The nitrile group of the compound of Example 192 is reduced to the amine using standard catalytic hydrogenation with palladium 20 in acidic media (usually 0.01 N to 1 N HC1), which is then converted to the sulfamide using sulphamoyl chloride.

In this example a compound of the present invention is prepared wherein "X" is COOH and "Y" is NH by reacting the 25 methyl amine intermediate prepared in the previous example with chloroacetic acid.

In this example a compound of the present invention is prepared wherein "X" is PO(OH)2 and "Y" is NH by reacting 30 the methyl amine intermediate prepared in Example 206 with diethylchloromethyl phosphonate, and reacting the resulting product with trimethylsilylbromide.

In this example a compound of the present inventionis prepared wherein "X" is S02NH2 and "Y" is oxygen by reacting the alcohol intermediate prepared in Example203 with sulphamoyl chloride.

In this example a compound of the present inventionis prepared wherein R1 is H, R2 is phenyl, R3 and R4 are hydrogen, m is 0, n is 1, p is 0, and X is CN. A

10modification of the procedure disclosed in Mu-I11 et Lim, al. , J. Orcr. Chem. , Vol. 44, No. 22, 3826 (1979)ed.
is us A mixture of the compound of Example 184 and dimethylformamide dimethyl acetal is reacted at oom r temperature for two days and then evaporated to in dryness 15vacuo. The residue is crystallized to give the pureN-(dimethylamino)methylene derivative, which is cyclizedith w saturated methanolic ammonia to give the desired end product.

20In this example a compound of the present inventionis prepared wherein R1 is OCH3, R2 is phenyl, R3 and are hydrogen, m is 0, n is 1, p is 0, and X is CN. Usingthe compound B of Example 187, the S-methyl group is zed oxidi to methylsulfone, which then is converted to the final 25methoxy compound by treatment with sodium methoxidein methanol.

In this example a compound of the present invention is prepared wherein X is tetrazole. The compound of Example 30 192 is treated with lithium azide in the presence of ammonium chloride as a catalyst in dimethylformamide (DMF) at 100 degrees C to give the desired tetrazole.

;~~'; "."'?~ '~' r ... :~

In this example a compound of the present invention is prepared wherein X is triazole. The compound of Example 198 is treated with hydrazine hydrate to give the corresponding hydrazide, which is then treated with imino ether to give the desired triazole.

The compound prepared in Example 189 is tested for enzyme inhibition activity as in Example 188. At 1 mM phosphate the IC50 is 0.012 ~,M and at 50 mM phosphate the IC50 is 2.0 f~M .

In this example an amidine compound of the present invention is prepared, i.e., wherein X in the recited generic formula is CNHNH2. The compound A from Example 187 is reacted with sodium methoxide in methanol at room temperature for about 2 days to give a methyl-imidate intermediate. The intermediate is. then reacted with ammonia in methanol to give the amidine product.

Claims (22)

CLAIMS:
1. A compound 2-amino-7- (R) -3H, 5H-pyrrolo [3, 2-d] -pyrimidin-4-one wherein R is cyclohexenyl, cyclohexyl, or-CH2-R1, and wherein Rl is an optionally substituted 5- or 6-membered saturated heteroalicyclic group having oxygen, nitrogen or sulfur as heteroatom, optionally substituted pyridinyl, or optionally substituted C5-10 alicyclic group.
2. The compound of claim 1 wherein R is -CH2-R1 and R1 is optionally substituted 2-, 3-, or 4-piperidinyl, 2- or 3-tetrahydrofuranyl, 2- or 3-tetrahydrothienyl, 2- or 3-pyrrolidinyl, 2-, 3-, or 4-tetrahydropyranyl, or 2-, 3-, 4-pyridinyl.
3. The compound of claim 1 wherein R is optionally substituted cyclohexyl or 1-, 2-, or 3-cyclohexenyl.
4. The compound of claim 1 wherein R is -CH2-R1 and R1 is optionally substituted cyclopentyl, cyclohexyl, cycloheptyl, 1- or 2-adamantyl, 1- or 2-noradamantyl, 1-norbornanyl, 2-exo-norbornanyl, 2-endo-norbornanyl, 1- or 2-bicyclo[2.2.2]octanyl, 1-, 2-, 3-, 6-, or 8-bicyclo[3.2.1]octanyl, 1-, 2-, 3-, or 9-bicyclo[3.3.1]nonanyl, or 1- or 2-norbornenyl.
5. A compound of the formula wherein R- is H, NH2, or OCH3, R2 is an optionally substituted cyclic group of 5-7 carbon atoms optionally containing one or more heteroatoms, R3 and R4 are independently H or C1-4 alkyl, m is 0-4, n is 0-6, p is 0-1, X is CN, CSNH2, PO (OH)2, COON, SO2NH2, NH2, OH, CNHNH2, tetrazole, or triazole, COR5 where R5 is C1-4 alkyl, CF3, NH2, or OC1-4 alkyl, and Y is O or NH.
6. The compound of claim 5 wherein R1 is NH2, R3 and R4 are H, m is 0 and n is 1.
7. The compound of claim 5 or 6 wherein R2 is phenyl.
8. The compound of claim 5, 6 or 7 wherein X is CN, COOH, or CONH2.
9. The compound of claim 5 or 6 wherein R2 is an optionally substituted 5- or 6-membered aromatic or heteroaromatic group or an optionally substituted alicyclic group or heteroalicyclic group of 5-9 members.
10. A pharmaceutical composition for the selective suppression of T-cell function without diminishing humoral immunity, comprising an effective amount of a compound according to any one of claims 1 to 9, and a pharmaceutically acceptable excipient or carrier.
11. A pharmaceutical composition for treating or preventing a disease responsive to the inhibition of phosphorylis and metabolic breakdown of antiviral or antitumour purine nucleosides, comprising an effective amount of a compound according to any one of claims 1 to 9, and a pharmaceutically acceptable excipient or carrier.
12. A pharmaceutical composition for potentiating the effect of antiviral or antitumour purine nucleosides, comprising an effective amount of a compound according to any one of claims 1 to 9, and a pharmaceutically acceptable excipient or carrier.
13. Use of a compound according to any one of claims 1 to 9, for the suppression of T-cell function without diminishing humoral immunity.
14. Use of a compound according to any one of claims 1 to 9, for the treatment or prevention of a disease responsive to the inhibition of phosphorylis and metabolic breakdown of antiviral or antitumour purine nucleosides.
15. Use of a compound according to any one of claims 1 to 9. for potentiating the effect of antiviral or antitumour purine nucleosides.
16. Use of a compound according to any one of claims 1 to 9, in conjunction with an antiviral or antitumour purine nucleoside for the treatment or prevention of an HIV related illness.
17. A method for making methyl-3-amino-4-(2-cyano-1-cyclo-ethyl)-1H-pyrrole-2-carboxylate comprising the steps of:
a) reacting an optionally substituted cyclic aldehyde with cyanoacetic acid in the presence of ammonium acetate to make a 3-cyclo-substituted pentanedinitrile;
b) reacting the 3-cyclo-pentanedinitrile with an alkyl formats and a base to make a 3-cyclo-2-formylpentanedinitrile;
c) reacting the 3-cyclo-2-formylpentanedinitrile with glycine methyl aster hydrochloride and sodium or ammonium acetate to make methyl N-[(3-cyclo-2,4-dicyano)-2-butenyl]glycine;
d) reacting the methyl N-[(3-cyclo-2,4-dicyano)-2-butenyl]glycine with an alkyl chloroformate and DBN or DBU
to make methyl 3-amino-4-(2-cyano-1-cyclo-ethyl)-1-ethyl-1H-pyrrole-1,2-dicarboxylate; and e) reacting the methyl 3-amino-4-(2-cyano-1-cyclo-ethyl)-1-ethyl-1H-pyrrole-1,2-dicarboxylate with a base.
18. A method for making a mixture of 3-cyclo-3-[2-amino-4-oxo-3H-5H-pyrrolo[3,2-d]pyrimidin-7-yl]propanenitrile and 3-cyclo-3-[2-methylmercapto-4-oxo-3H,5H-pyrrolo[3,2-d]pyrimidin-7-yl]propanenitrile comprising the steps of:
f) reacting methyl 3-amino-4-(2-cyano-1-cyclo-ethyl)-1H-pyrrole-2-carboxylate with benzoylisothiocyanate to make N-benzoyl-N'-[4-(2-cyano-1-cyclo-ethyl)-2-methoxycarbonyl-1H-pyrrol-3-yl]thiourea;
g) reacting N-benzoyl-N'-(4-(2-cyano-1-cyclo-ethyl)-2-methoxycarbonyl-1H-pyrrol-3-yl]thiourea with an alkyl halide to make N-benzoyl-N'-[4-(2-cyano-1-cyclo-ethyl)-2-methoxycarbonyl-1H-pyrrol-3-yl]S-methylthiourea; and h) reacting N-benzoyl-N'-[4-(2-cyano-1-cyclo-ethyl)-2-methoxycarbonyl-1H-pyrrol-3-yl]-S-methylthiourea with methanolic or ethanolic ammonia.
19. A method for making 3-cyclo-3-[2-methoxy-4-oxo-3H,5H-pyrrolo[3,2-d]pyrimidin-7-yl]propanenitrile comprising the steps of:
i) reacting 3-cyclo-3-[2-methylmercapto-4-oxo-3H,5H-pyrrolo[3,2-d]pyrimidin-7-yl]propanenitrile with an oxidizing agent to make 3-cyclo-3-[2-methylsulfonyl-4-oxo-3H,5H-pyrrolo[3,2-d]pyrimidin-7-yl]propanenitrile; and j) reacting the 3-cyclo-3-[2-methylsulfonyl-4-oxo-3H,5H-pyrrolo[3,2-d]pyrimidin-7-yl]propanenitrile with sodium alkoxide.
20. A method for making 3-cyclo-3-[4-oxo-3H,5H-pyrrolo[3,2-d]pyrimidin-7-yl]propanenitrile comprising the steps of:
f) reacting methyl 3-amino-4-(2-cyano-1-cyclo-ethyl)-1H-pyrrole-2-carboxylate with dimethylformamide dimethyl acetal to make methyl 4-(2-cyano-1-cyclo-ethyl)-3-[N-(dimethylaminomethylene)amino]-1H-pyrrole-2-carboxylate; and g) reacting the methyl 4-(2-cyano-1-cyclo-ethyl)-3-[N-(dimethylaminomethylene)amino]-1H-pyrrole-2-carboxylate with methanolic ammonia.
21. The method of claim 17 wherein the cyclic substituent is phenyl, 2- or 3-thienyl, 2- or 3-furanyl, 2-, 3-, or 4-pyridinyl, 2- or 3-pyrrolyl, 2-, 4-, or 5-thiazolyl, 2- or 3-pyrazinyl, 3- or 4-pyridazinyl, 3-, 4-, or 5-pyrazolyl, 1- or 2-adamantyl, cyclopentyl, cyclohexyl, cycloheptyl, or morpholinyl.
22. A compound of the formula wherein R1 and R2 are as defined in claim 5, 6 or 7.
CA002072123A 1989-10-31 1990-10-12 Inhibitors of purine nucleoside phosphorylase Expired - Fee Related CA2072123C (en)

Applications Claiming Priority (11)

Application Number Priority Date Filing Date Title
US429,097 1982-09-30
US429,100 1989-10-31
US07/429,098 US4985434A (en) 1989-10-31 1989-10-31 7-substituted derivatives of 2-amino-3H,5H-pyrrolo(3,2-d)pyrimidin-4-ones and pharamceutical uses and compositions containing the same
US429,099 1989-10-31
US07/429,097 US5008270A (en) 1989-10-31 1989-10-31 2-amino-7-(heterocyclomethyl)-3H,5H-pyrrolo[3,2-d]pyrimidin-4-ones and pharmaceutical uses and compositions containing the same
US07/429,100 US4985433A (en) 1989-10-31 1989-10-31 2-amino-7-(pyridinylmethyl)-3H,5H-pyrrolo[3,2-d]pyrimidin-4-ones and pharmaceutical uses and compositions containing the same
US429,098 1989-10-31
US07/429,099 US5008265A (en) 1989-10-31 1989-10-31 2-amino-7-(alicyclomethyl)-3H,5H,-pyrrolo[3,2-d]pyrimidin-4-ones and pharmaceutical uses and compositions containing the same
US44279889A 1989-11-29 1989-11-29
US442,798 1989-11-29
PCT/US1990/005756 WO1991006548A1 (en) 1989-10-31 1990-10-12 Inhibitors of purine nucleoside phosphorylase

Publications (2)

Publication Number Publication Date
CA2072123A1 CA2072123A1 (en) 1991-05-01
CA2072123C true CA2072123C (en) 2001-06-12

Family

ID=27541578

Family Applications (1)

Application Number Title Priority Date Filing Date
CA002072123A Expired - Fee Related CA2072123C (en) 1989-10-31 1990-10-12 Inhibitors of purine nucleoside phosphorylase

Country Status (9)

Country Link
EP (1) EP0500653A4 (en)
JP (1) JP2866478B2 (en)
AU (1) AU654264B2 (en)
CA (1) CA2072123C (en)
FI (1) FI103972B1 (en)
HU (1) HUT61765A (en)
LV (1) LV10100B (en)
NO (1) NO301423B1 (en)
WO (1) WO1991006548A1 (en)

Families Citing this family (14)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5726311A (en) * 1989-11-29 1998-03-10 Biocryst Pharmaceuticals, Inc. 7-disubstituted-methyl-4-oxo-3H,5H-pyrrolo 3,2-d!pyrimidine and pharmaceutical uses and compositions containing the same
US5236926A (en) * 1992-02-03 1993-08-17 Warner-Lambert Company 9-substituted-8-halo or -8-hydroxy-9-deazaguanines as inhibitors or PNP for pharmaceutical compositions
HUT71559A (en) * 1992-04-21 1995-12-28 Biocryst Pharm Inc 4-oxo-3h,5h-pyrrolo[3,2-d]pyrimidine and pharmaceutical compositions containing them
US5736549A (en) * 1994-10-05 1998-04-07 Chiroscience Limited Hypoxanthine and guanine compounds
GB9520364D0 (en) * 1995-10-05 1995-12-06 Chiroscience Ltd Compouundds
GB9520363D0 (en) * 1995-10-05 1995-12-06 Chiroscience Ltd Compounds
AR012871A1 (en) * 1997-05-29 2000-11-22 Novartis Ag 2-AMINO-7- (2-HYDROXYETHYL-1-SUBSTITUTED) -3,5-DIHYDROPYROL (3,2-D) PIRIMIDIN -4-ONAS, PHARMACEUTICAL COMPOSIONS AND THE USE OF THE SAME FOR THE MANUFACTURING OF A MEDICINAL PRODUCT.
US6174888B1 (en) 1998-05-28 2001-01-16 Novartis Ag 2-amino-7-(1-substituted-2-hydroxyethyl)-3,5-dihydropyrrolo[3,2-D]pyrimidin-4-ones
JP4496586B2 (en) * 2000-01-24 2010-07-07 日産化学工業株式会社 Process for producing quinolylacrylonitrile and its intermediate
CN100379750C (en) * 2002-08-21 2008-04-09 阿尔伯爱因斯坦医科叶希瓦大学 Nucleoside phosphorylase and nucleosidase inhibitors
NZ523970A (en) * 2003-02-04 2005-02-25 Ind Res Ltd Process for preparing inhibitors of nucleoside phoshorylases and nucleosidases
US8138191B2 (en) 2007-01-11 2012-03-20 Critical Outcome Technologies Inc. Inhibitor compounds and cancer treatment methods
EP2225226B1 (en) 2007-12-26 2016-08-17 Critical Outcome Technologies, Inc. Compounds and their use in a method for treatment of cancer
EP2552915B1 (en) 2010-04-01 2017-07-19 Critical Outcome Technologies Inc. Compounds for the treatment of hiv

Family Cites Families (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPS5988484A (en) * 1982-11-09 1984-05-22 Takeda Chem Ind Ltd Novel 7-deazapurine derivative
CA1293727C (en) * 1986-08-26 1991-12-31 Catherine Rose Kostlan 9-deazaguanines
CA1294960C (en) * 1986-10-24 1992-01-28 Thomas C. Malone 7-deazaguanines as immunomodulators
US4921858A (en) * 1986-10-24 1990-05-01 Warner-Lambert Company 7-deazaguanines as immunomodulators
DE3726044A1 (en) * 1987-02-11 1988-08-25 Bayer Ag ACARICIDES ON THE BASIS OF AZOMETHINE DERIVATIVES AND AZOMETHINE OF 2,3-DIAMINOMALEAINSAURENITRILE
US4927830A (en) * 1988-04-08 1990-05-22 The Regents Of The University Of Michigan Acyclic pyrrolo[2,3-D]pyrimidine analogs as antiviral agents
HU217870B (en) * 1989-02-27 2000-04-28 Biocryst Pharmaceuticals Inc. 9-substituted 9-dezaza-guanine derivatives, pharmaceutical compositions containing them and process for the preparation thereof
US4985433A (en) * 1989-10-31 1991-01-15 Biocryst, Inc. 2-amino-7-(pyridinylmethyl)-3H,5H-pyrrolo[3,2-d]pyrimidin-4-ones and pharmaceutical uses and compositions containing the same
US4985434A (en) * 1989-10-31 1991-01-15 Biocryst, Inc. 7-substituted derivatives of 2-amino-3H,5H-pyrrolo(3,2-d)pyrimidin-4-ones and pharamceutical uses and compositions containing the same

Also Published As

Publication number Publication date
JPH05504551A (en) 1993-07-15
NO301423B1 (en) 1997-10-27
JP2866478B2 (en) 1999-03-08
NO921679L (en) 1992-06-17
FI103972B (en) 1999-10-29
LV10100A (en) 1994-05-10
LV10100B (en) 1995-08-20
NO921679D0 (en) 1992-04-29
EP0500653A1 (en) 1992-09-02
CA2072123A1 (en) 1991-05-01
FI921900A0 (en) 1992-04-28
HUT61765A (en) 1993-03-01
HU9201450D0 (en) 1992-09-28
WO1991006548A1 (en) 1991-05-16
FI921900L (en) 1992-04-28
AU654264B2 (en) 1994-11-03
EP0500653A4 (en) 1995-08-23
AU6712590A (en) 1991-05-31
FI103972B1 (en) 1999-10-29

Similar Documents

Publication Publication Date Title
CA2072123C (en) Inhibitors of purine nucleoside phosphorylase
JP4205168B2 (en) Pyrimidine derivatives and process for producing the same
US5712303A (en) Pyrazoles and pyrazolopyrimidines having CRF antagonistic activity
US6441018B2 (en) Pyrazoles and pyrazolopyrimidines having CRF antagonistic activity
LT3448B (en) Inhibitors of purine nucleoside phosphorylase and process for preparing thereof
HU193799B (en) Process for producing 3-acyl-amino-methyl-imidazo-square bracket opened-1,2-a-square bracket closed-pyridine derivatives and pharmaceutical compositions containing them
KR101446680B1 (en) Thienopyrimidinone derivatives as mGluR1 antagonists
KR20000069605A (en) Pyrrolo-[3,4-d]Pyrimidinone Derivatives and Their Use as Medicaments
WO1992014707A1 (en) Poly-4-aminopyrrole-2-carboxyamide derivatives, processes for their preparation and pharmaceutical compositions which contain them
US5565463A (en) 9-subtituted-8-unsubstituted-9-deazaguanines
US5189039A (en) 7-disubstituted-methyl-4-oxo-3H,5H-pyrrolo[3,2d]pyrimidine and pharmaceutical uses and compositions containing the same
KR101081446B1 (en) 2-substituted phenyl-5,7-dialkyl-3,7-dihydropyrrole [2,3-d] pyrimidin-4-one derivatives, their preparation and pharmaceutical uses
EP0638080A1 (en) 7-DISUBSTITUTED-METHYL-4-OXO-3H -,5H --PYRROLO 3,2-d -]PYRIMIDINE AND PHARMACEUTICAL USES AND COMPOSITIONS CONTAINING THE SAME
US4985433A (en) 2-amino-7-(pyridinylmethyl)-3H,5H-pyrrolo[3,2-d]pyrimidin-4-ones and pharmaceutical uses and compositions containing the same
JPH04273878A (en) Heterocyclic compound and preparation thereof
RU2097384C1 (en) 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
US5726311A (en) 7-disubstituted-methyl-4-oxo-3H,5H-pyrrolo 3,2-d!pyrimidine and pharmaceutical uses and compositions containing the same
EP1400522A1 (en) New compounds for treating impotence
Su et al. Pyrimidines. 21. Novel reactions of 5‐cyano‐1, 3‐dimethyluracil with carbon nucleophiles. A facile preparation of certain pyrido [2, 3‐d] pyrimidines
US6103900A (en) Pyrazoles and pyrazolopyrimidines having CRF antagonistic activity
WO2000053581A2 (en) Heterocyclic compounds having antitumor activity
US6946554B2 (en) Process for the preparation of 9-deazaguanine derivatives
EP0816359B1 (en) 4-Amino-2-ureido-pyrimidine-5-carboxamides, processes for their preparation, medicaments containing these compounds, and their use
KR20020010669A (en) Salts of Cis-[4-Cyano-4-(3-Cyclopentyloxy-4-Methoxyphenyl) Cyclohexan-1-Carboxylate]
JP2001521483A (en) Method for producing 9-deazaguanine derivative

Legal Events

Date Code Title Description
EEER Examination request
MKLA Lapsed