JP2003511454A - Pyrrolo [2,3-d] pyrimidine nucleoside analogs - Google Patents

Abstract

(57) SUMMARY Provided are compositions and methods for pyrrolo [2,3-d] pyrimidine nucleoside analogs having substituents at the C4 'and C5' positions of the ribofuranose moiety. The envisaged compositions exhibit, inter alia, anti-cancer and immunomodulatory effects with low cytotoxicity.

Description

Detailed Description of the Invention

[0001]     (Field of the invention)   The field of the invention is nucleoside analogues.

BACKGROUND OF THE INVENTION Nucleoside analogs have been used for many years as antimetabolites for the treatment of cancer and viral infections. After entering the cell, nucleoside analogs are often phosphorylated by the nucleoside salvage pathway, in which analogs are typically phosphorylated to the corresponding mono-, di- and triphosphates. Among other intracellular applications, triphosphorylated nucleoside analogs are often used as substrates for DNA or RNA polymerases, and consequently incorporated into DNA or RNA. If the triphosphorylated nucleoside analogues are strong polymerase inhibitors, they can induce early termination of nascent nucleic acid molecules. Disruption of gene expression or function can occur when triphosphorylated nucleoside analogs are incorporated into nucleic acid replication or transcripts.

At the cellular level, nucleoside analogues can also interfere with the cell cycle, and a particularly desirable effect of nucleoside analogues involves the induction of apoptosis of cancer cells. In addition, nucleoside analogues are also known to alter certain immune responses.

Various nucleoside analogues are known in the art with relatively potent anti-cancer effects. Known agents include thymidylate synthase inhibitors such as 5-fluorouridine, adenosine deaminase inhibitors including 2-chloroadenosine, and neplanocin A, an inhibitor of S-adenosyl homocysteine hydrolase. However, all or almost all of the known nucleoside analogues also carry the threat to normal mammalian cells, mainly because these nucleoside analogues lack the proper selectivity between normal and tumor cells. Unfortunately, the lack of adequate selectivity is often associated with serious side effects and therefore often limits the potential for treatment with such analog compounds.

There are a variety of nucleoside analogs known in the art, but all or almost all of them have one or more disadvantages. Therefore, there remains a need to provide improved methods and compositions for nucleoside analogs.

DISCLOSURE OF THE INVENTION The present invention is directed to nucleoside analogs having modifications on the sugar moieties of the pyrrolo [2,3-d] pyrimidine nucleoside analogs, wherein such modifications are toxic to the mammalian cells. Can be significantly reduced, while at the same time giving significant cytotoxicity to cancer cells. These modifications are due to C4 in the ribofuranose moiety.
Includes, but is not limited to, substitutions at the'and C5 _' positions. The present invention also provides that some pyrrolo [2,3-d] pyrimidine nucleoside analogs, including potentiation of type 1 cytokines such as IL-2 and suppression of type 2 cytokines such as IL-4, are desirable. It is clarified that it has an immunoregulatory effect. These immunomodulatory properties
It is considered to be useful in the treatment of anti-cancer drugs, anti-viral drugs and autoimmune diseases, inflammation and prevention of graft rejection.

[0007]   In one aspect of the invention, the nucleoside analogue compound has the formula (I):

[0008]

[Chemical 6] [Wherein A is O, S or CH_TwoAnd X is H, NH_TwoOr OH and Y is H
, Halogen or NH_TwoAnd Z is H, halogen, R, OH, OR, SH, S
R, NH_Two, NHR, NR_Two, CN, C (O) NH_Two, COOH, COOR, C
H_TwoNH_Two, C (= NOH) NH_Two, And C (= NH) NH_TwoSelected from the group consisting of
R is alkyl, alkenyl, alkynyl or aralkyl, R is_Two And R_ThreeAre independently selected from the group consisting of H, F and OH, R_FourIs hydrogen,
R is selected from the group consisting of alkyl, alkenyl, alkynyl and aralkyl, R _Four Optionally has at least one of a heteroatom and a functional group, R₅Is H,
OH, OP (O) (OH)_Two, P (O) (OH)_Two, OP (O) (OR ')_Twoor
Is P (O) (OR ')_Two, R'is a masking group, and R_{5 '}Is
Selected from the group consisting of alkyl, alkenyl, alkynyl and aralkyl,
R_{5 '}Has at least 2 carbon atoms, and optionally heteroatoms and functional groups
Have at least one of A pyrrolo [2,3-d] pyrimidine nucleoside analogue having a structure according to
is there.

[0009]   In another aspect of the invention, the nucleoside analogue compound has the formula (II):

[0010]

[Chemical 7] [In the formula, Z is CN, C (O) NH ₂ , C (= NH) NH ₂ or C (= NOH) N
H ₂ and R ₄ and R _{5 ′} are independently selected from the group consisting of hydrogen, alkyl, alkenyl, alkynyl and aralkyl, R ₄ and R _{5 ′} are independently and optionally a heteroatom and A pyrrolo [2,3-d] pyrimidine nucleoside having a structure according to [1], which contains at least one of the functional groups, provided that R ₄ and R _{5 ′} are not both hydrogen.

In a further aspect of the invention, the envisaged compounds are used to inhibit tumor growth or regulate the production of type 1 and type 2 cytokines and chemokines.

Various objects, features, aspects and advantages of the present invention will become more apparent from the following detailed description of the preferred embodiments of the present invention, taken in conjunction with the accompanying drawings.

BRIEF DESCRIPTION OF THE FIGURES FIG. 1 is a first exemplary synthetic scheme of the reactions involved in the preparation of compounds according to the present invention. FIG. 2 is a second exemplary synthetic scheme of the reactions involved in the preparation of compounds according to the present invention. FIG. 3 is a third exemplary synthetic scheme of the reactions involved in the preparation of compounds according to the present invention. FIG. 4 is a fourth exemplary synthetic scheme of the reactions involved in the preparation of compounds according to the present invention. FIG. 5 is a fifth exemplary synthetic scheme of the reactions involved in the preparation of compounds according to the present invention. FIG. 6 is a sixth exemplary synthetic scheme of the reactions involved in the preparation of compounds according to the present invention. FIG. 7 shows exemplary compounds according to the present invention. 8A and 8B are graphs showing the effect of the compounds according to the present invention on the expression of type 1 and type 2 cytokines, respectively. FIG. 9 is a table showing the cytotoxicity of various compounds according to the present invention. FIG. 10 is a table showing rates of DNA synthesis in cells treated with various compounds according to the present invention. FIG. 11 shows V from human prostate cancer cells when treated with compounds according to the invention.
3 is a graph showing inhibition of EGF release. Figure 12: I from human prostate cancer cells when treated with compounds according to the invention
Figure 7 is a graph showing inhibition of L-8 release.

DETAILED DESCRIPTION OF THE INVENTION Pyrrolo [2,3-d] pyrimidine nucleoside analogues according to formulas (I) and (II) exert various biological effects on normal and hyperproliferative cells. Was recognized.

[0015]

[Chemical 8] [Wherein A is O, S or CH_TwoAnd X is H, NH_TwoOr OH and Y is H
, Halogen or NH_TwoAnd Z is H, halogen, R, OH, OR, SH, S
R, NH_Two, NHR, NR_Two, CN, C (O) NH_Two, COOH, COOR, C
H_TwoNH_Two, C (= NOH) NH_Two, And C (= NH) NH_TwoSelected from the group consisting of
R is alkyl, alkenyl, alkynyl or aralkyl, R is_Two And R_ThreeAre independently selected from the group consisting of H, F and OH, R_FourIs hydrogen,
R is selected from the group consisting of alkyl, alkenyl, alkynyl and aralkyl, R _Four Optionally has at least one of a heteroatom and a functional group, R₅Is H,
OH, OP (O) (OH)_Two, P (O) (OH)_Two, OP (O) (OR ')_Twoor
Is P (O) (OR ')_Two, R'is a masking group, and R_{5 '}Is
Selected from the group consisting of alkyl, alkenyl, alkynyl and aralkyl,
R_{5 '}Has at least 2 carbon atoms, and optionally heteroatoms and functional groups
Have at least one of. ]   As used herein, "alkyl", "alkenyl", "alkynyl" and "a
It should be especially appreciated that the term "rualkyl" refers to both straight and branched types.
Substituent R_TwoAnd R_ThreeRegarding R_TwoAnd R_ThreeBoth independently pair α or β planes.
You should be aware that you can be an elephant. Furthermore, C_{5 '}The above substitutions are non-identical
C₅The above substitutions may result in R or S chiral centers. When used here
The term "heteroatom" refers to atoms other than carbon in organic molecules and is
Terror atoms include halogen, nitrogen, oxygen and sulfur. When used here, "sensual
The term “group” refers to a reactive bond (eg, a double or triple bond) or a reactive group (eg, —OH).
, -SH, -NH_Two, -N_Three, -CN, -COOH, -CHO, -CONH_Two,
And so on).

Particularly contemplated pyrrolo [2,3-d] pyrimidine nucleoside analogues are:
Wherein Z is CN, C (O) NH ₂ or C (═NH) NH ₂ , R _{5 ′} has at least 2 carbon atoms and is selected from the group consisting of alkyl, alkenyl, alkynyl and aralkyl. According to formula (I).

A pyrrolo [2,3-d] pyrimidine nucleoside analogue according to formula (II) has the following structure:

[0018]

[Chemical 9] [In the formula, Z is CN, C (O) NH ₂ , C (= NH) NH ₂ or C (= NOH) N
H ₂ and R ₄ and R _{5 ′} are independently selected from the group consisting of hydrogen, alkyl, alkenyl, alkynyl and aralkyl, R ₄ and R _{5 ′} are independently and optionally a heteroatom and Containing at least one of the functional groups, provided that R ₄ and R _{5 ′} are not both hydrogen, and the remaining substituents are as defined in formula (I).

However, the compounds according to the invention may also take forms and dosage forms other than those mentioned above, in particular envisaged forms in which the prodrug form or the envisaged molecule is
Chemically and / or enzymatically modified to improve pharmacological and / or pharmacodynamic properties, including higher specificity for target organ, cellular or non-cellular fractions and longer half-life in vivo Including modified forms.

For example, cholesterol adducts can be formed to increase target specificity for the liver, or apolipoprotein adducts formed to enhance the penetration of modifying agents across the blood-brain barrier into the brain. can do. In another example, the receptor-ligand complex can be synthesized to target the modifying agent to specific cells that express the ligand-specific receptor. Alternatively, the antibody or antibody fragment complex can be formed to enhance selective delivery of the modifying agent to non-cellular sites. There are many prodrugs and modified forms known in the art, and particularly contemplated prodrug forms are US Provisional Application 60/216418 filed April 17, 2000, which is incorporated herein by reference. Including the prodrugs mentioned in the issue. In a further example, charged or uncharged groups, lipophilic or polar groups can be added to the putative molecule to increase its half-life in serum or in other target organs and / or cells. It should be appreciated that in further examples it is also possible to di- or tri-phosphorylate putative compounds which are phosphorylated at the C ₅ atom, or to incorporate thiophosphates.

While it is generally preferred that the envisaged compound have a sugar moiety in the D configuration, the compound is L
It is also envisioned to have a sugar moiety in the configuration. Further stereochemical aspects include, among others:
The substituents in the envisaged compounds may be in the α or β phase, optionally including the R and S configurations at the C ₅ atom.

The compounds envisioned include a variety of agents, including liquid, syrup or gel forms (eg for injection, oral ingestion or topical administration) and solid forms (eg oral intake, injection or deposition in body cavities). It can be formulated into molds. For example, if a compound according to the present invention is assumed to be unstable in the gastric environment, preferably infusion of an isotonic solution is particularly contemplated. Alternatively, intranasal application or inhalation in liquid form may be appropriate to avoid acid degradation. On the other hand, if the envisaged compound is known to be resistant to digestive degradation, the envisaged form may be administered in the form of a syrup or tablet. The envisaged compound may also be formulated for topical or transdermal application, depending on the particular application. There are many more dosage forms known in the art, all of which are envisioned as suitable in the context of the invention described herein, and particularly contemplated dosage forms are:
Donald C. Monkhouse and Christopher T. Rho
Des. (edit); ISBN: 082479852X, "Drug Products for Clinical Trials: Drug Products, Manufacturing, and Quality Control Intellectual Guidance (Drug Products f
or Clinical Trials: An Intl. Guide to
Formulation, Production, Quality Control
ol) ”.

It should be further appreciated that the envisioned compounds and dosage forms may include functional and non-functional additives. For example, a skin permeation enhancer can be added if transdermal drug delivery is desired. Alternatively, agents including bacteriostatic, antiviral or immunomodulatory agents may be added to synergistically or additively enhance the function of the putative compound. Examples of non-functional additives include fillers, antioxidants, flavoring agents or colorants to enhance the specific quality of the envisaged compound.

Regarding the expected concentration of the compound, the concentration is about 1 μM when measured at the site of action.
Is preferably in the range of about 100 μM. However, suitable concentrations may also be in the range of 999 nM to 10 nM and below, especially if the affinity of the envisaged compound is below 1 μM. On the other hand, if the putative compound exhibits a relatively short half-life or has a high turnover, then the putative concentration may be 0.1 mM to 100 mM and higher. Thus, while the dosage of the envisioned compound can vary significantly, the appropriate dosage can be readily determined in vitro or in animal studies.

Among the various biological actions of 5 ′ and 4 ′ modified pyrrolopyrimidine nucleoside analogues, particularly important biological actions are regulation of type 1 and type 2 cytokine production, regulation of neoplastic conditions. (Eg inhibition of DNA synthesis or inhibition of cell proliferation), and inhibition of chemokine and growth factor release as described below.

Accordingly, a postulated method of altering the secretion of cytokines from a cell comprises providing a compound according to formula (I), at a concentration of formula (I) effective to alter the secretion of cytokines. The method may further include contacting the cell with the compound.
Although all possible combinations of substituents in formula (I) are generally considered, especially envisaged compounds are those in which R ₄ and R _{5 ′} consist of hydrogen, alkyl, alkenyl, alkynyl and aralkyl. Independently selected from the group, R ₄ and R _{5 ′} independently and optionally containing at least one of a heteroatom and a functional group, the remaining substituents being defined above in formula (I) Is a compound according to formula (I) In an alternative aspect, the compound used to alter the secretion of cytokines from cells also has the structure:

[0027]

[Chemical 10] [In the formula, Z is CN, C (O) NH ₂ , C (= NH) NH ₂ , C (= NNH ₂ ) N
H ₂ or —C (= NOH) NH ₂ , and R ₅ is H, OH, OP (O) (OH
) ₂ , P (O) (OH) ₂ , OP (O) (OR ') ₂ or P (O) (OR') ₂ and R'is a masking group].

[0028] Envisioned cytokines are especially type 1 (eg IFNγ) and type 2 (eg IL).
-4) Contains cytokines. With respect to cells, all cells known to produce and / or secrete cytokines are suitable, but particularly contemplated cells include lymphocytes and cancer cells (eg prostate cancer cells, see below).

In a further aspect of the invention, a method of inhibiting the proliferation of hyperproliferative cells is of formula (I)
And a second step of contacting the hyperproliferative cells with a concentration of the compound effective to inhibit proliferation of the hyperproliferative cells. Particularly preferred compounds are those wherein R ₄ is selected from the group consisting of hydrogen, alkyl, alkenyl, alkynyl and aralkyl, R ₄ optionally containing at least one of a heteroatom and a functional group, R _{5 ′} Is selected from the group consisting of hydrogen, alkyl, alkenyl, alkynyl and aralkyl, R _{5 ′} has at least 2 carbon atoms and optionally contains at least one of a heteroatom and a functional group,
Provided that R ₄ and R _{5 ′} are not hydrogen atoms, and the remaining substituents are of the formula (I
) And a compound according to formula (I) as defined above.

Particularly contemplated hyperproliferative cells include cancer cells, and particularly contemplated cancer cells are prostate cancer cells. While not wishing to be bound by any particular theory, suppression of proliferation includes suppression of DNA synthesis.

In a further aspect of the invention, a method of inhibiting the release of growth factors from cells comprises providing a compound according to formula (I), and a concentration of the effective concentration for inhibiting the release of growth factors. It is contemplated to include another step of contacting the cell with the compound. Although the release of various growth factors can be suppressed by the methods presented here,
Inhibition of VEGF release is of particular concern. Similarly, although all cells known to secrete growth factors are envisioned in connection with the methods presented herein, specifically contemplated cells include cancer cells, particularly prostate cancer cells.

With respect to the synthesis of envisioned compounds, the pyrrolo [2,3-d] pyrimidine nucleoside analogs according to the present invention can be synthesized through a variety of synthetic routes,
The following procedure is provided as an example only.

Synthesis of C _5′- Modified Pyrrolo [2,3-d] pyrimidine Nucleoside Analogues 5′-substituted nucleoside analogues are pyrrolo [2,3-d] pyrimidine bases and appropriately protected 5′-substituted ribofuranose. Prepared from the condensation of As shown in FIG. 1, published procedures (Jones et al., “Methods in Carbohydrate Chemistry (Me
"thods in carbohydrate Chemistry""(Wh
istler and Moffat), Volume VI, p. 315-322, Acad
Compound 1 prepared according to emic Press, New York, (1972) is treated with various nucleophiles such as Grignard reagent to produce Compound 2, which is benzoylated and then treated with trifluoroacetic acid. Compound 4 was obtained. Benzoylation followed by treatment with acetic anhydride / acetic acid in the presence of sulfuric acid yielded compound 6, which was used for condensation with pyrrolo [2,3-d] pyrimidine base.

Compound 7 prepared according to published procedures (Jones et al., “Methods in carbohydrate Chem.
(Istry) "(edited by Whistler and Moffat), Volume VI, p. 31
5-322, Academic Press, New York, (1972).
Was converted to a tosylate derivative, which was reduced with lithium aluminum hydride to obtain compound 8. Compound 8 was converted to compound 9 by a procedure similar to that shown in FIG.
Converted to. 9 and pyrrolo [2,3-d] pyrimidine 19 as shown in Scheme 2
2 and subsequent transformations provided compounds 10-15 as illustrated in FIG.

As shown in FIG. 3, the compound 2 was converted to the sulfonate 16 and subjected to nucleophilic substitution to obtain the compound 17 with the reversed configuration. Deprotection of isopropylidene followed by acetylation gave tetraacetate 18. The condensation of a 5C-substituted, protected ribofuranose with a nucleoside base is shown in FIG. Published procedure (T
olman et al. Org. Chem. (1969, 91, 1022-2108)
5-Cyanopyrrolo [2,3-d] pyrimidine 19 prepared according to
The compound 6 was condensed with compound 6 in the presence of trimethylsilyl triflate by a similar procedure as described for sides Nucleotides 1993, 12, 643-648). The resulting coupling product was subjected to debromination via hydrogenation to give compound 20. Treatment of 20 with ammonia in anhydrous methanol gave compounds 21 and 23. Compound 21 was oxidized to give compound 22. Compound 23
Was converted to the carboxamide derivative 24. Compounds 23 and 24 were oxidized to give compound 25. Compound 25 was treated with hydroxyamine to yield compound 26, which was hydrogenated over Raney nickel to give 27. Alternatively, compound 27 was also prepared by heating compound 25 with ammonia in a pressure bomb.

Synthesis of C4 ′ Modified Pyrrolo [2,3-d] pyrimidine Nucleoside Analogs In FIG. 5, compound 1 is treated with formaldehyde in aqueous sodium hydroxide to produce the 4′-hydroxymethyl derivative 28, which Was selectively protected to give compound 29. Subsequent protection with DMT and removal of TBS produced compound 31. Compound 31 can be converted into various substituents. The 4-C-substituted derivative, which underwent a similar conversion to the 5-C-substituted ribofuranose (Scheme 1), can be converted to compound 35, which is used for condensation with the nucleoside base.

Similar to the C _{5 ′} substituted pyrrolopyrimidine nucleoside analogues, condensation of compound 35 with compound 19 as shown in FIG. 6 provides 4 ′ substituted analogue 36. Subsequent conversion can give the 4'-C substituted pyrrolopyrimidine nucleosides 37-42.

Synthesis of 2'Modifications and Other Pyrrolo [2,3-d] pyrimidine Nucleoside Analogs The following pyrrolopyrimidine nucleoside analogs were prepared for biological testing. Some have been published (shown as known compounds) but they are shown in FIG. Known compounds 43, 44, 52-55 and 57 are published procedures (Hinshaw et al., J. Org. Chem. 1970, 92, 236-241.
). Compound 56 was prepared by hydrogenation of compound 52. Known Compound 49 (Krawczyk et al., Nucleosides Nucleoti)
Des 1989, 8.97-115) was treated with sodium nitrite to give compound 5
I got 0. Known compounds 45 and 48 were prepared according to published procedures (Ramasamy et al., J. He.
terocyclic Chem. 1988, 25, 1043-1046). Compound 45 was treated with ammonia-methanol to yield compound 46 and hydrogenated to yield compound 47. Compound 58-63 is compound 52-5
Prepared from compound 45 by a procedure similar to that used for 7. Known Compound 64 (Krawczyk et al., Nucleosides Nucleotide
s 1989, 8.97-115) was converted to compounds 65-67. Known compound 68 (Ramasamy et al., Tetrahedron 1986, 42, 586).
9-5878) was converted to compounds 69 and 70.

Preparation and activation of human T cells in vitro Peripheral blood mononuclear cells were isolated from healthy donors by density gradient centrifugation and then L
ymphokwick (One Lambda, Canonga Park CA)
Were used to enrich T cells. Purified T cells are> 99% CD2 +, <1% HLA-D
R + and <5% CD25 +, RPMI-AP5 (5% autologous plasma, 1% L-
Glutamine, 1% penicillin / streptomycin and 0.05% 2-mercaptoethanol in RPMI1640 medium). For the determination of cytokine protein levels, 2 ng phorbol myristate acetate and ionomycin 0.
1 mg (PMA-ION, both Calbiochem, San Diego
, CA) and incubated for 48 hours at 37 ° C. in 96 well plates in the presence of various guanosine nucleosides 0 or 10 μM to activate T cells (0.2 × 10 6 cells in 0.2 ml volume). . After activation, supernatants were analyzed for cell-derived cytokine production.

Extracellular Cytokine Assays ELISA kit specific for IFNγ and IL-4 (Biosource, C
amarillo, CA) was used to measure human cytokine levels in cell supernatants after appropriate dilution. All ELISA results were expressed in pg / ml.

Pyrrolo [2,3-d on extracellular cytokine levels in activated human T cells
] Action of Pyrimidine Nucleoside Analog Compound Pyrrolo [2,3-d] pyrimidine Nucleoside Analog Compound 0 or 10 μM is 1
8A and 8 for individual human donors, the effect of type IV cytokines, IFNγ and type 2 cytokines, IL-4, on PMA / ionomycin stimulated T cell expression.
It is shown in B. Cytokine levels were measured in cell-free supernatant by ELISA. The most potent action is 7-b-D-ribofuranyl-4-oxopyrrolo-[[
It was observed for 2,3-d] pyrimidine-5-carboxamidine. This compound potentiated activated IL-4 production by 498% ± 83 and suppressed IFNγ by 43% ± 4 compared to activated control levels of each cytokine. Data are presented as the percentage of activated control, calculated as the ratio of activated T cell cytokine levels in the presence of test nucleoside to untreated activated T cell cytokine levels x 100%. Zero effects on cytokine levels by the tested nucleosides represent a 100% activation control value percentage. Absolute levels of PMA-ION-induced cytokine secretion (pg / ml ± standard deviation) were 22954 ± 3391 for IFNγ and 162 ± 40 for IL-4. Resting levels were <30 pg / ml for all cytokines tested.

Cytotoxicity of Pyrrolo [2,3-d] pyrimidine Nucleoside Analogs In Vitro The pyrrolo [2,3-d] pyrimidine nucleoside analogs of the present invention exhibit some level of cytotoxicity in vitro, It is bioactive. In these studies, test compounds were tested on normal human fibroblasts, human prostate cancer cells 81, human melanoma cells 140, human lung cancer cells 177, and human ovarian cancer cells R and NR (all ATCs).
(Available from C). In these experiments, cells were plated at a density of 2000 cells / 200 μl medium / well (96-well plate). Immediately after seeding the cells, the test compound was applied once to the wells in the concentration range of 0.78-100 μM. Colorimetric cytotoxicity assay MTS was performed 72 hours after treatment. The EC50 was calculated based on the collected readings and is shown in FIG. Some compounds are 100
Concentrations below μM show lack of cytotoxicity. In such cases, EC50>
It is expressed as 100. In other cases, the EC50 indicates the concentration of test compound required to damage 50% of the cell population.

Pyrrolo [2,3-d] pyrimidine Nucleoside Analogs Dose-Dependently Inhibit DNA Synthesis in Cells Cultured In Vitro Pyrrolo [2,3-d] pyrimidine nucleoside analogs are dependent on the level of DNA. Inhibits the growth of human cells cultured in vitro when measured. The experimental conditions were the same as described above. Compounds were applied once and DNA levels were measured after 72 hours. At that point, half of the medium was removed from the culture well and replaced with pure water. The cells were then transferred to -70 ° C for at least 12 hours. In the next step, the cells were returned to room temperature from -70 ° C and Hoechst 33342 1 µM was added to each well. After incubation for 2 hours, the fluorescence signal (360-530n
m) was measured. According to this method, the formed DNA-Hoechst 33
Due to the presence of the 342 complex, the fluorescence intensity is proportional to the amount of DNA. The result is shown in FIG.
Shown in. The numbers are D compared to the amount of DNA at the start of the experiment (2 hours after cell inoculation).
Represents a multiple of the increase in NA quantity. 7 in untreated prostate cancer cells and normal cells
The DNA levels were increased 5.78 and 4.47 fold, respectively, during the 2 hour culture.

Compounds 23a (5′-R) and 23a (5′-S) inhibit the secretion of VEGF from human prostate cancer cells in vitro Compounds 23a (5′-R) and 23a (5′-S) Is a human prostate cancer cell, H
It strongly inhibits the secretion of vascular endothelial growth factor (VEGF) from TB81. V
EGF is recognized as an angiogenesis marker because it is essential for endothelial cell migration and proliferation and microangiogenesis in vivo. To clarify this, 0.5
× 10 ⁵ cells were plated in 5 ml of medium in Petri dishes with a diameter of 10 cm. The compounds were applied for 72 hours immediately after planting. After that, the medium is collected and VEG
VEGF levels were measured using the F Elisa Assay (R & D Systems) and expressed as pg of VEGF / ml of medium. The results are shown in Fig. 11. According to these results, both compounds dose-dependently inhibit VEGF secretion.

Compounds 23a (5′-R) and 23a (5′-S) inhibit IL-8 release from human prostate cancer cells cultured in vitro Compounds 23a (5′-R) and 23a (5) '-S) is human prostate cancer cell, H
It shows an inhibitory effect on the secretion of interleukin-8 (IL-8) from TB81. IL-8 belongs to the class of chemoattractant cytokines (alpha type) and is involved in neutrophil-induced inflammatory processes. Chemokines are generally known to be produced by various types of cancer. Inhibition of chemokine production by cancer cells has been shown to be beneficial to the host in several studies. To demonstrate the potential of these two compounds to inhibit IL-8 secretion from prostate cancer cells, prostate cancer cells HTB81 were labeled in vitro with the concentrations of 5'-R and 5'-S configurations shown in the graph. Treated with compound 23a. The medium collected from the culture is used as R & D S
IL-8 levels were analyzed using the IL-8 Elisa Assay from systems. According to the results obtained, as shown in FIG.
The secretion of L-8 can be inhibited in a dose-dependent manner.

However, it should be appreciated that the biological effects of the envisioned compounds need not be limited to the particular effects described above. In particular, the compounds according to the invention generally have a cytostatic effect in various hyperproliferative disorders, including localized and / or metastatic cancers (eg lymphomas and carcinomas), benign prostatic hyperplasia and keratosis. It is considered to indicate. Although the inventor has observed substantial biological effects on IL-4 (type 2 cytokine) and IFNγ (type 1 cytokine), in general, compounds according to the invention are
It is considered to be biologically active in the regulation of cytokines other than L-4 and IFNγ. In particular, it is believed that the compounds may increase or decrease the expression / secretion of individual cytokines or sets of cytokines. Therefore, it is believed that the compounds according to the present invention may modulate the immune system of the body to achieve a more pronounced type 1 or type 2 response. As a result, the compounds according to the invention are able to act on biological systems either directly by acting as inhibitors of viral polymerases and / or indirectly by activating the immune system to specific humoral or cellular responses. It is thought to be effective in reducing the titer of the virus in. Moreover, the compounds according to the invention are also believed to be useful in reducing the response of the immune system to allografts or xenografts by reducing the strength of the cellular response to allografts or xenografts. To be

EXAMPLES The following protocols set forth exemplary syntheses of various compounds in accordance with the present invention and are intended to be illustrative only of the inventive concepts presented herein and not intended to be limiting thereof. do not do.

[0048]   Methyl 2,3-O-isopropylidene-5 (R, S) -C-ethynyl-β-ly
Preparation of bofuranoside (2b)   Methyl 4-C, 5-O-didehydro-2,3-O-isopropylidene-β-D
-Ribofuranoside (Jones et al., "Methods in Carbohydrate Chemistry (Method
in Carbohydrate Chemistry) ", Volume I, p.
315-322 (1972), 4.00 g, 19.78 mmol) under argon.
To a stirred solution in anhydrous THF (20 mL) at −42 ° C., ethynyl magnesium bromide
(0.5 M in 80 mL THF, 40 mmol) was added dropwise. After addition, raw
The stirred mixture was gently warmed to 0 ° C. (˜90 minutes). Ice (50g) / water (5
(0 mL) was added to quench the reaction, and the mixture was stirred for 30 minutes. 10% acetic acid aqueous solution
After neutralizing with, the mixture was extracted twice with ethyl acetate. The combined organic phase is dried (Na _Two SO_Four), Concentrated. Chromatography on silica (ethyl acetate-hexa
1: 4) to give 3.48 g of the title compound (R / S ratio 1: 1) as a white solid.
Obtained. The following compounds were similarly prepared: Methyl 4-C, 5-O-didehydro
-2,3-O-isopropylidene-β-D-ribofuranoside and ethyl bromide
From sium, methyl 2,3-O-isopropylidene-5 (R) -C-methyl-β
-D-ribofuranoside (2a) was prepared. Methyl 4-C, 5-O-didehydro
-2,3-O-isopropylidene-β-D-ribofuranoside and vinyl bromide
From sium, methyl 2,3-O-isopropylidene-5 (R) -C-vinyl-β
-D-ribofuranoside (2c) was prepared. Methyl 4-C, 5-O-didehydro
-2,3-O-isopropylidene-β-D-ribofuranoside and arylmagnesium bromide
From sium, methyl 5 (R) -C-allyl-2,3-O-isopropylidene-β
-D-ribofuranoside (2d) was prepared.

Methyl 2,3-O-isopropylidene-5-O-methanesulfonyl-5 (R)
Preparation of -C-methyl-β-D-ribofuranoside (16) Methyl 2,3-O-isopropylidene-5 (R) -C-methyl-β-D-ribofuranoside (2a, 7.24 g, 33.17 mmol). Anhydrous pyridine (
Methanesulfonyl chloride (3.1 mL, 39.92 mmo to a stirred solution in 50 mL).
l) was added. The resulting mixture was stirred at room temperature for 1 hour, cooled to 0 ° C. and washed with water (1.
(0 mL) was added to stop the reaction, and the mixture was stirred at room temperature for 30 minutes. The solvent was evaporated, the residue dissolved in ethyl acetate, washed 3 times with brine, dried (Na ₂ SO ₄ ).
, Concentrated. Chromatography on silica (30% EtOAc in hexane) gave 8.62 g of the title compound 16 as a colorless syrup.

Preparation of methyl 2,3-O-isopropylidene-5-O-acetyl-5 (S) -C-methyl-β-D-ribofuranoside (17) Methyl 2,3-O-isopropylidene-5 O-methanesulfonyl-5 (R)
-C-methyl-β-D-ribofuranoside (16, 8.62 g, 29.1 mmol)
) And NaOAc (anhydrous, 3.5 g, 42.5 mmol) in anhydrous DMF (350
The stirred suspension in (mL) was heated at 125 ° C. under argon for 4 days. The solvent was evaporated and the residue was chromatographed on silica (25% EtOAc in hexane) to give 4.0 g of the title compound 17 as a white solid.

Methyl 2,3-O-isopropylidene-4-C-hydroxymethyl-β-D-
Preparation of Ribofuranoside (28) Methyl 4-C, 5-O-didehydro-2,3-O-isopropylidene-β-D
-Ribofuranoside 1 (20.22 g, 100 mmol) in dioxane (3
To a stirred solution in 80 mL) formaldehyde (37% solution, 76 mL) was added dropwise, followed by 2M NaOH (188 mL). The resulting reaction mixture was stirred at room temperature for 2
Stir 0 h, cool to 0 ° C., neutralize (10% acetic acid), concentrate (˜50%).
, Extracted twice with methylene chloride. The combined organic phases were dried (Na ₂ SO ₄ ) and concentrated to dryness. Chromatography on silica (4% methanol in chloroform) gave 20.2 g of the title compound 28 as a white solid.

[0052]   Methyl 2,3-O-isopropylidene-5-deoxy-β-D-ribofuranosi
Preparation of code (8)   Methyl 2,3-O-isopropylidene-β-D-ribofuranoside (14.2 g
, 70.0 mmol) in a stirred solution of anhydrous pyridine (250 mL) at 10 ° C.
P-toluenesulfonyl chloride (19.1 g, 100 mL) in small portions (in 30 minutes
Added). The resulting reaction mixture was stirred at room temperature for 18 hours, cooled to 0 ° C.
Water (5.0 mL) was added to stop the reaction, and the mixture was stirred at room temperature for 30 minutes. Steam the solvent
I fired. The residue was dissolved in ethyl acetate, washed 3 times with brine, dried (Na _Two SO_Four), Concentrated and dried. Chromatography on silica (ethyl acetate-hex
Sun 1: 3) gave 24.1 g of the title compound as a white solid.

LiAlH ₄ (4.58 g 120.5 mmol) in anhydrous diethyl ether (1
To a stirred suspension in 20 mL), diethyl ether-toluene (2.5: 1, 140
Methyl 2,3-O-isopropylidene-5-Op-toluenesulfonyl-β-D-ribofuranoside (13.1 g, 36.55 mmol) in (mL) was added.
The resulting mixture was refluxed for 22 hours, cooled to room temperature, diluted with ethyl acetate (25 mL) and quenched with water (5.0 mL). The solvent was evaporated. The residue was dissolved in ethyl acetate, washed 3 times with brine, dried (Na ₂ SO ₄ ) and concentrated to dryness. Chromatography on silica (ethyl acetate-hexane 1: 3) gave 3.58 g of the title compound as a colorless liquid.

Preparation of methyl 5 (R) -C-allyl-5-O-benzoyl-2,3-O-isopropylidene-β-D-ribofuranoside (3d) Methyl 5 (R) -C-allyl-2, 3-O-isopropylidene-β-D-ribofuranoside (4.49 g, 18.38 mmol) in anhydrous pyridine (40 m
L) Benzoyl chloride (2.7 mL, 23.0 mmol) was added to the stirred solution in L).
The resulting mixture was stirred at room temperature for 18 hours, cooled with ice, quenched by the addition of water (1 mL) and stirred at room temperature for 30 minutes. The solvent was evaporated, the residue dissolved in ethyl acetate, washed 3 times with brine, dried (Na ₂ SO ₄ ) and concentrated. Chromatography on silica (12% ethyl acetate in hexane) gave 6.26 g of the title compound 3d as a colorless syrup. The following compounds were prepared similarly: Methyl 5 (R, S) -C-ethynyl-2,3-O-isopropylidene-β-D
-Ribofuranoside (2b) to methyl 5-O-benzoyl-5 (R, S) -C-
Ethynyl-2,3-O-isopropylidene-β-D-ribofuranoside (3b, R
/ S ratio: 1: 1) was prepared. Methyl 2,3-O-isopropylidene-4-C-
Methyl 4-C-benzoyloxymethyl-5-O-benzoyl-2,3-O-isopropylidene-β-D-ribofuranoside was prepared from hydroxymethyl-β-D-ribofuranoside.

Preparation of methyl 5 (R) -C-allyl-5-O-benzoyl-β-D-ribofuranoside (4d) Methyl 5 (R) -C-allyl-5-O-benzoyl-2,3-O TF of isopropylidene-β-D-ribofuranoside (3d, 6.2 g, 17.8 mmol)
_{A-H 2} O mixture (9: 1) was treated stirred for 90 min at 0 ° C., concentrated to dryness at 0 ° C.. The residue was dissolved in a methanol-toluene mixture (20 mL, 1: 1) and concentrated to dryness. Chromatography on silica (ethyl acetate-hexane 1: 1) gave 3.70 g of the title compound 4d as a white solid. The following compounds were similarly prepared: methyl 5-O-benzoyl-5 (R, S) -C-ethynyl-2,
3-O-isopropylidene-β-D-ribofuranoside (3b) to methyl 5-O
-Benzoyl-5 (R, S) -C-ethynyl-β-D-ribofuranoside (4b,
An R / S ratio of 1: 1) was prepared. Methyl 5-O-benzoyl-4-C-benzoyloxymethyl-2,3-O-isopropylidene-β-D-ribofuranoside to methyl 5-O-benzoyl-4-C-benzoyloxymethyl-β-D-ribofuranoside Was prepared.

Methyl 5 (R) -C-allyl-2,3,5-tri-O-benzoyl-β-D
-Preparation of ribofuranoside (5d) Methyl 5 (R) -C-allyl-5-O-benzoyl-β-D-ribofuranoside (4d, 3.60 mg, 11.68 mmol) in anhydrous pyridine (80 mL).
) Benzoyl chloride (3.0 mL, 25.84 mmol) was added to the stirred solution.
The resulting mixture was stirred at room temperature for 18 hours, cooled with ice, quenched by the addition of water (1 mL), then stirred at room temperature for 30 minutes. The mixture was concentrated, diluted with ethyl acetate, washed 3 times with brine, dried (Na ₂ SO ₄ ) and concentrated to dryness. Chromatography on silica (15% ethyl acetate in hexane) gave 5.3 g of the title compound 5d as a colorless syrup. The following compounds were similarly prepared:
From methyl 5-O-benzoyl-5 (R, S) -C-ethynyl-β-D-ribofuranoside (4b) to methyl 5 (R, S) -C-ethynyl-2,3,5-tri-O.
-Benzoyl-β-D-ribofuranoside (5b, R / S ratio: 1: 1) was prepared. Methyl 4-C-benzoyloxymethyl-5-O-benzoyl-β-D-ribofuranoside to methyl 4-C-benzoyloxymethyl-2,3,5-tri-
O-benzoyl-β-D-ribofuranoside was prepared.

Preparation of 1-O-methyl-2,3,5-tri-O-benzoyl-5 (R) -C-vinyl-β-D-ribofuranose (5c) Methyl 2,3-O-isopropylidene -5 (R) -C-Vinyl-β-D-ribofuranoside (2c, 1.0 g, 4.3 mmol) in trifluoroacetic acid and water (9
The solution in the mixture of 1: 1, v / v, 11 mL) was stirred at 0 ° C. for 30 minutes and concentrated to dryness. The residue was dissolved in methanol, concentrated to dryness (3 times), then dissolved in pyridine and evaporated, and finally dissolved in anhydrous pyridine (11 mL). Benzoyl chloride (1.9 mL, 16 mmol) was added to this solution. The reaction mixture was stirred at 25 ° C. for 16 hours and poured into ice water (20 mL). Mix the mixture with dichloromethane (20 mL
) And the organic phase was dried over sodium sulfate and concentrated to dryness. The residue was chromatographed on silica (0-5% ethyl acetate in dichloromethane), 1
． 0 g of the title compound 5c was obtained as a syrup.

Preparation of 1-O-acetyl-2,3,5-tri-O-benzoyl-5 (R) -C-allyl-β-D-ribofuranose (6d) Methyl 5 (R) -C-allyl -2,3,5-tri-O-benzoyl-β-D
-Ribofuranoside (5d, 4.0 g, 7.74 mmol) acetic acid (14 m
L) and acetic anhydride (1.75 mL, 18.37 mmol) in a stirred solution in concentrated sulfuric acid (
200 μL, 3.79 mmol in 4.0 mL acetic acid) were added. The resulting mixture was stirred at room temperature for 20 hours, cooled to 0 ° C., diluted with cold ethyl acetate, water, 5% Na.
Wash with aqueous HCO ₃ and brine, dry (Na ₂ SO ₄ ) and concentrate. Chromatography on silica (ethyl acetate-hexane 1: 4) gave 2.82 g of the title compound 6d (α / β ratio 1: 2) as a colorless foam. The following compounds were prepared similarly: Methyl 5 (R, S) -C-ethynyl-2,3,5-t
ri-O-benzoyl-β-D-ribofuranoside (5b) to 1-O-acetyl-5 (R, S) -C-ethynyl-2,3,5-tri-O-benzoyl-β-D
Ribofuranose (6b, R / S ratio: 1: 1 and α / β ratio: 1: 2) was prepared. Methyl 4-C-benzoyloxymethyl-2,3,5-tri-O-benzoyl-β-D-ribofuranoside to 1-O-4-C-benzoyloxymethyl-2
, 3,5-tri-O-benzoyl-D-ribofuranose (α / β ratio: 1: 3)
Was prepared. Methyl 2,3-O-isopropylidene-5 (R) -C-methyl-β
From -D-ribofuranoside to 5 (R) -C-methyl-1,2,3,5-tetra
-O-Acetyl-β-D-ribofuranose was prepared. From methyl 5-O-acetyl-2,3-O-isopropylidene-5 (R) -C-methyl-β-D-ribofuranoside to 1,2,3,5-tetra-O-acetyl-5 (S)- C-methyl-
D-ribofuranose (6a) was prepared. Methyl 5-O-acetyl-2,3-O
From isopropylidene-β-D-ribofuranoside to 5-deoxy-1,2,3-
Tri-O-acetyl-D-ribofuranose (9) was prepared. Methyl 2,3
5-tri-O-benzoyl-5 (R) -C-vinyl-β-D-ribofuranoside to 1-O-acetyl-2,3,5-tri-O-benzoyl-5 (R) -C-
Vinyl-β-D-ribofuranose (6c) was prepared.

4-amino-6-bromo-5-cyano-7- (2,3,5-tri-O-benzoyl-5 (R) -C-allyl-β-D-ribofuranosyl) pyrrolo [2,3 -D
] Preparation of Pyrimidine 4-Amino-6-bromo-5-cyanopyrrolo [2,3-d] pyrimidine (To
lman et al. Org. Chem. 1969, 91, 1022-2108, 1
． 05 g, 4.41 mmol) and ammonium sulfate (50 mg) HMDS (
75 mL) and a suspension in anhydrous m-xylene (25 mL) was refluxed under argon for 18 hours. The solvent was evaporated and the residue dried under vacuum. The residue is anhydrous 1,2-
Dissolve in dichloroethane (80 mL) and 1-O-acetyl-2,3,5-tri
-O-benzoyl-5 (R) -C-allyl-D-ribofuranose (2.00 g,
3.67 mmol). While cooling with ice, TMSOTf (1.3 mL
, Anhydrous 1,2-dichloroethane (7.30 mmol in 5 mL) was added. The mixture was stirred under argon at room temperature for 30 minutes, then refluxed for 90 hours, quenched it by pouring it (cold) into ice / NaHCO ₃ (50 mL) and filtering. The organic phase was separated, dried (Na ₂ SO ₄ ) and concentrated. Chromatography on silica (Et
OAc-hexane 2: 3) gave 1.81 g of the title compound as a colorless solid. The following compounds were similarly prepared: 1-O-acetyl-2,3,5-t
From ri-O-benzoyl-5 (R, S) -C-ethynyl-D-ribofuranose and 4-amino-6-bromo-5-cyanopyrrolo [2,3-d] pyrimidine, 4
-Amino-6-bromo-5-cyano-7- (2,3,5-tri-O-benzoyl-5 (R, S) -C-ethynyl-β-D-ribofuranosyl) pyrrolo [2,3-
d] Pyrimidine (R / S ratio: 1: 1) was prepared. From 1-O-acetyl-4-benzoyloxymethyl-2,3,5-tri-O-benzoyl-D-ribofuranose and 4-amino-6-bromo-5-cyanopyrrolo [2,3-d] pyrimidine, 4-Amino-6-bromo-5-cyano-7- (4-benzoyloxymethyl-2,3,5-tri-O-benzoyl-β-D-ribofuranosyl) pyrrolo [2
, 3-d] pyrimidine was prepared. 1,2,3,5-tetra-O-acetyl-5 (R) -C-methyl-D-ribofuranose and 4-amino-6-bromo-5
-Cyanopyrrolo [2,3-d] pyrimidine to 4-amino-6-bromo-5-
Cyano-7- (1,2,3,5-tetra-O-acetyl-5 (R) -C-methyl-β-D-ribofuranosyl) pyrrolo [2,3-d] pyrimidine was prepared.
From 1,2,3,5-tetra-O-acetyl-5 (S) -C-methyl-D-ribofuranose and 4-amino-6-bromo-5-cyanopyrrolo [2,3-d] pyrimidine, 4 -Amino-6-bromo-5-cyano-7- (1,2,3,5-t
Etra-O-acetyl-5 (S) -C-methyl-β-D-ribofuranosyl) pyrrolo [2,3-d] pyrimidine was prepared. From 1,2,3-tri-O-acetyl-5-deoxy-D-ribofuranose and 4-amino-6-bromo-5-cyanopyrrolo [2,3-d] pyrimidine to 4-amino-6-bromo- 5-cyano-
7- (2,3-di-O-acetyl-5-deoxy-β-D-ribofuranosyl)
Pyrrolo [2,3-d] pyrimidine was prepared. 1-O-acetyl-2,3,5-
From tri-O-benzoyl-5 (R) -C-vinyl-β-D-ribofuranose and 4-amino-6-bromo-5-cyanopyrrolo [2,3-d] pyrimidine, 4
-Amino-6-bromo-5-cyano-7- (2,3,5-tri-O-benzoyl-5 (R) -C-vinyl-β-D-ribofuranosyl) pyrrolo [2,3-d] pyrimidine Was prepared.

4-amino-5-cyano-7- (2,3,5-tri-O-benzoyl-5 (
Preparation of (R) -C-allyl-β-D-ribofuranosyl) pyrrolo [2,3-d] pyrimidine (20e) 4-amino-6-bromo-5-cyano-7- (2,3,5-tri- O-benzoyl-5 (R) -C-allyl-β-D-ribofuranosyl) pyrrolo [2,3-d
] To a solution of pyrimidine (738 mg, 1.0 mmol) in acetic acid (25 mL) was added 2
Zinc dust (1.04 g, 16.0 mmol) in 1 portion (at intervals of 1 hour)
Was added. The reaction mixture was stirred at room temperature for 20 hours and filtered. The filtrate is evaporated to dryness,
The residue was chromatographed on silica (ethyl acetate-hexane 1: 1).
), 450 mg of the title compound 20e was obtained as a colorless foam. The following compound was similarly prepared: 4-amino-6-bromo-5-cyano-7- (2,3,5-t
ri-O-benzoyl-5 (R, S) -C-ethynyl-β-D-ribofuranosyl) pyrrolo [2,3-d] pyrimidine to 4-amino-5-cyano-7- (2,3
, 5-tri-O-benzoyl-5 (R, S) -C-ethynyl-β-D-ribofuranosyl) pyrrolo [2,3-d] pyrimidine (R / S ratio: 1: 1) (20b) was prepared. did. 4-amino-6-bromo-5-cyano-7- (2,3,5-tri-
O-benzoyl-5 (R) -C-vinyl-β-D-ribofuranosyl) pyrrolo [2
, 3-d] pyrimidine to 4-amino-5-cyano-7- (2,3,5-tri
-O-benzoyl-5 (R) -C-vinyl-β-D-ribofuranosyl) pyrrolo [
2,3-d] pyrimidine (20c) was prepared.

4-amino-5-cyano-7- (2,3,5-tri-O-benzoyl-5 (
R) -C-Propyl-β-D-ribofuranosyl) pyrrolo [2,3-d] pyrimidine (20f) 4-amino-6-bromo-5-cyano-7- (2,3,5-tri- O-benzoyl-5 (R) -C-allyl-β-D-ribofuranosyl) pyrrolo [2,3-d
] Pyrimidine (400 mg, 0.54 mmol) and 10% Pd / C (100 m
g, ~ 50% water) dioxane (50 mL) and triethylamine (0.5 mL)
) Was shaken in a hydrogenator (H ₂ , 20 psi) for 4 hours. The catalyst was filtered and washed (dioxane). The combined filtrate was concentrated and the residue was chromatographed on silica (ethyl acetate-hexane 1: 1) to give the title compound 20f.
340 mg were obtained as a colorless foam. The following compound was similarly prepared: 4-amino-6-bromo-5-cyano-7- (2,3,5-tri-O-benzoyl-
5 (R, S) -C-ethynyl-β-D-ribofuranosyl) pyrrolo [2,3-d]
From pyrimidine to 4-amino-5-cyano-7- (2,3,5-tri-O-benzoyl-5 (R, S) -C-ethyl-β-D-ribofuranosyl) pyrrolo [2,3
-D] pyrimidine (R / S ratio: 1: 1) (20d) was prepared. 4-amino-6
-Bromo-5-cyano-7- (4-benzoyloxomethyl-2,3,5-tr
i-O-benzoyl-β-D-ribofuranosyl) pyrrolo [2,3-d] pyrimidine to 4-amino-5-cyano-7- (4-benzoyloxomethyl-2,3,3
5-tri-O-benzoyl-β-D-ribofuranosyl) pyrrolo [2,3-d]
Pyrimidine was prepared. 4-amino-6-bromo-5-cyano-7- (1,2,
3,5-tetra-O-acetyl-5 (R) -C-methyl-β-D-ribofuranosyl) pyrrolo [2,3-d] pyrimidine to 4-amino-5-cyano-7- (
1,2,3,5-tetra-O-acetyl-5 (R) -C-methyl-β-D-
Ribofuranosyl) pyrrolo [2,3-d] pyrimidine (20a) was prepared. 4-
Amino-6-bromo-5-cyano-7- (1,2,3,5-tetra-O-acetyl-5 (S) -C-methyl-β-D-ribofuranosyl) pyrrolo [2,3-d
] From pyrimidine to 4-amino-5-cyano-7- (1,2,3,5-tetra
-O-acetyl-5 (S) -C-methyl-β-D-ribofuranosyl) pyrrolo [2
, 3-d] pyrimidine (20a) was prepared. 4-Amino-6-bromo-5-cyano-7- (1,2,3-tri-O-acetyl-5-deoxy-β-D-ribofuranosyl) pyrrolo [2,3-d] pyrimidine to 4-amino -5-cyano-7
-(1,2,3-tri-O-acetyl-5-deoxy-β-D-ribofuranosyl) pyrrolo [2,3-d] pyrimidine was prepared. 4-amino-5-cyano-7
-(2,3-dideoxy-β-D-pent-2-enofuranosyl) pyrrolo [2,2
3-d] pyrimidine to 4-amino-5-cyano-7- (2,3-dideoxy-
β-D-glycero-pentofuranosyl) pyrrolo [2,3-d] pyrimidine was prepared.

Preparation of 4-amino-5-cyano-7- (5 (R) -C-allyl-β-D-ribofuranosyl) pyrrolo [2,3-d] pyrimidine (23e) 4-amino-5-cyano -7- (2,3,5-tri-O-benzoyl-5 (
A solution of (R) -C-allyl-β-D-ribofuranosyl) pyrrolo [2,3-d] pyrimidine (300 mg, 0.454 mmol) in methanol (40 mL) at 0 ° C was saturated with ammonia. The solution was left at room temperature for 2 days. The solvent was evaporated and the residue was suspended in DMF (20 mL) with NaOAc (anhydrous, 20 mg). The mixture was stirred under argon at 120 ° C. for 5 hours. The solvent was evaporated. The residue was adsorbed on silica gel and eluted from the silica gel column (methanol-ethyl acetate 1:25) to give the title compound (145 mg) as a colorless solid.

The product contains two main compounds 21 and 23, which can be separated by chromatography on silica gel and then heated in DMF. Compound 21 was prepared through this procedure. The following compounds were similarly prepared: 4
-Amino-5-cyano-7- (2,3,5-tri-O-benzoyl-5 (R)
-C-propyl-β-D-ribofuranosyl) pyrrolo [2,3-d] pyrimidine to 4-amino-5-cyano-7- (5 (R) -C-propyl-β-D-ribofuranosyl) pyrrolo [2 , 3-d] pyrimidi (23f) was prepared. 4-amino-5
4-Cyano-7- (2,3,5-tri-O-benzoyl-5 (R, S) -C-ethynyl-β-D-ribofuranosyl) pyrrolo [2,3-d] pyrimidine to 4-amino-5 -Cyano-7- (5 (R, S) -C-ethynyl-β-D-ribofuranosyl) pyrrolo [2,3-d] pyrimidine (RS ratio: 1: 1) (23b) was prepared. 4-Amino-5-cyano-7- (2,3,5-tri-O-benzoyl-5
(R, S) -C-Ethyl-β-D-ribofuranosyl) pyrrolo [2,3-d] pyrimidine to 4-amino-5-cyano-7- (5 (R, S) -C-ethyl-β- D
-Ribofuranosyl) pyrrolo [2,3-d] pyrimidine (RS ratio: 1: 1) (2
3d) was prepared. 4-Amino-5-cyano-7- (4-benzoyloxomethyl-2,3,5-tri-O-benzoyl-β-D-ribofuranosyl) pyrrolo [
4-Amino-5-cyano-7- (4-hydroxymethyl-β-D-ribofuranosyl) pyrrolo [2,3-d] pyrimidine (33d) was prepared from 2,3-d] pyrimidine. 4-amino-5-cyano-7- (1,2,3,5-tetra-O-
Acetyl-5 (R) -C-methyl-β-D-ribofuranosyl) pyrrolo [2,3-
d] pyrimidine to 4-amino-5-cyano-7- (5 (R) -C-methyl-β
-D-ribofuranosyl) pyrrolo [2,3-d] pyrimidine (23a (5'-R)
) Was prepared. 4-amino-5-cyano-7- (1,2,3,5-tetra-
O-acetyl-5 (S) -C-methyl-β-D-ribofuranosyl) pyrrolo [2,2
3-d] pyrimidine to 4-amino-5-cyano-7- (5 (S) -C-methyl-β-D-ribofuranosyl) pyrrolo [2,3-d] pyrimidine (23a (5′-
S)) was prepared. 4-amino-5-cyano-7- (1,2,3-tri-O-
Acetyl-5-deoxy-β-D-ribofuranosyl) pyrrolo [2,3-d] pyrimidine to 4-amino-5-cyano-7- (5-deoxy-β-D-ribofuranosyl) pyrrolo [2,3-d ] Pyrimidine (10) was prepared. 4-amino-5
Cyano-7- (2,3,5-tri-O-benzoyl-5 (R) -C-vinyl-
β-D-ribofuranosyl) pyrrolo [2,3-d] pyrimidine to 4-amino-5
-Cyano-7- (5 (R) -C-vinyl-β-D-ribofuranosyl) pyrrolo [2
, 3-d] pyrimidine (23c) was prepared.

4-amino-5-cyano-7- (2,3-di-O-methanesulfonyl-5-
O-tert-butyldiphenylsilyl-β-D-ribofuranosyl) pyrrolo [2
Preparation of, 3-d] pyrimidine Toyokamycin 43 (5.83 g, 20.0 mmol) was added to a stirred solution of anhydrous pyridine (100 mL) at 0 ° C. to give tert-butylchlorodiphenylsilane (6.2.
mL, 24.0 mmol) was added. The resulting mixture was stirred at room temperature for 18 hours, then cooled to 0 ° C. and methanesulfonyl chloride (3.4 mL, 44.0 mmol).
Was added. The resulting mixture was stirred at room temperature for 2 hours, cooled with ice, quenched by the addition of water (2 mL) and stirred at room temperature for 30 minutes. The solvent was evaporated. The residue was dissolved in ethyl acetate, washed 3 times with brine, dried (Na ₂ SO ₄ ) and concentrated. Chromatography on silica (ethyl acetate-hexane 3: 2) yielded 8.41 g of the title compound as a colorless solid.

4-amino-5-cyano-7- (5-O-tert-butyldiphenylsilyl-2,3-didehydro-2,3-dideoxy-β-D-ribofuranosyl) pyrrolo [2,3-d] Preparation of pyrimidine Tellurium powder (200 mesh, 640 mg, 5.0 mmol) was sealed under argon and lithium triethylborohydride (1.0 M in THF, 11.25 mL,
11.25 mmol). The mixture was stirred at room temperature for 6 hours, then cooled to 5 ° C. and 4-amino-5-cyano-7- (2,3-d in THF (12 mL).
i-O-methanesulfonyl-5-O-tert-butyldiphenylsilyl-β-
D-ribofuranosyl) pyrrolo [2,3-d] pyrimidine (1.40 g, 2.09
mmol) was added. The resulting mixture was stirred at room temperature for 18 hours, cooled with ice, water (0 ° C., 5 mL) was added to stop the reaction, and the mixture was stirred at room temperature for 30 minutes. The solvent was evaporated and the residue was extracted with ethyl acetate. The extracts were concentrated and the residue was chromatographed on silica (15% ethyl acetate in hexane) to give the title compound 640.
mg was obtained as a colorless foam.

4-amino-5-cyano-7- (2,3-didehydro-2,3-dideoxy-
Preparation of β-D-ribofuranosyl) pyrrolo [2,3-d] pyrimidine (49) 4-amino-5-cyano-7- (5-O-tert-butyldiphenylsilyl-2,3-didehydro-2,3 -Dideoxy-β-D-ribofuranosyl) pyrrolo [2,3-d] pyrimidine (2.55 g, 5.32 mmol) in anhydrous TH at 5 ° C.
To a stirred solution in F (100 mL) tetrabutylammonium fluoride (1.
0M, 6.6 mL) was added. The resulting mixture was stirred at room temperature for 3 hours and concentrated.
By chromatography on silica (6% methanol in ethyl acetate) 1.0
Obtained 9 g of the title compound 49 as a colorless solid.

Preparation of 5-cyano-7- (5 (R) -C-methyl-β-D-ribofuranosyl) pyrrolo [2,3-d] -4-pyrimidone (25a) 4-amino-5-cyano- 55 of 7- (5 (R) -C-methyl-β-D-ribofuranosyl) pyrrolo [2,3-d] pyrimidine (306 mg, 1.0 mmol)
Sodium nitrate (59 mL) was added to a stirred solution of water (30 mL) and acetic acid (2.0 mL) at 0 ° C.
0 mg, 8.55 mmol) was added in small portions. The resulting mixture was stirred at 70 ° C. for 3 hours and additional sodium nitrate (300 mg, 4.30 mmol) was added.
The mixture was stirred at the same temperature for a further 18 hours. The solvent was evaporated and the residue was chromatographed on silica (12% methanol in methylene chloride) to give 210 mg of the title compound 25a (5′-R) as a colorless solid. Similarly, the following compound was prepared: 4-amino-5-cyano-7- (5 (S) -C-methyl-β.
-D-ribofuranosyl) pyrrolo [2,3-d] pyrimidine to 4-amino-5-
Cyano-7- (5 (S) -C-methyl-β-D-ribofuranosyl) pyrrolo [2,2
3-d] -4-pyrimidone (25a (5'-S)) was prepared. 4-amino-5
-Cyano-7- (5-deoxy-β-D-arabinofuranosyl) pyrrolo [2,3
4-Amino-5-cyano-7- (β-D-arabinofuranosyl) pyrrolo [2,3-d] -4-pyrimidone (58) was prepared from -d] pyrimidine. 4-Amino-5-cyano-7- (5-deoxy-β-D-ribofuranosyl) pyrrolo [2,3
-D] pyrimidine to 4-amino-5-cyano-7- (5-deoxy-β-D-
Ribofuranosyl) pyrrolo [2,3-d] -4-pyrimidone (11) was prepared.
4-Amino-5-cyano-7- (2,3-dideoxy-β-D-pent-2-enofuranosyl) pyrrolo [2,3-d] pyrimidine to 4-amino-5-cyano-
7- (2,3-dideoxy-2,3-didehydro-β-D-glycero-pento-
Furanosyl) pyrrolo [2,3-d] -4-pyrimidone (50) was prepared. 4-
Amino-5-cyano-7- (2,3-dideoxy-β-D-glycero-pentofuranosyl) pyrrolo [2,3-d] pyrimidine to 4-amino-5-cyano-7-
(2,3-Dideoxy-β-D-glycero-pentofuranosyl) pyrrolo [2,3
-D] -4-Pyrimidone (65) was prepared. 4-amino-5-cyano-7- (
2-Deoxy-β-D-erythropentofuranosyl) pyrrolo [2,3-d] pyrimidine to 4-amino-5-cyano-7- (2-deoxy-β-D-furanosyl) pyrrolo [2,3 -D] -4-Pyrimidone (69) was prepared.

7- (5 (R) -C-methyl-β-D-ribofuranosyl) pyrrolo [2,3-d
] -4-Pyrimidone-5-carboxamidoxime (24a) Preparation 5-Cyano-7- (5 (R) -C-methyl-β-D-ribofuranosyl) pyrrolo [2,3-d] -4-pyrimidone ( 240 mg, 0.784 mmol), hydroxylamine hydrochloride (163 mg, 2.352 mmol), and potassium carbonate (1
A stirred suspension of 62 mg, 1.176 mmol) in ethanol (50 mL) was refluxed under argon for 18 hours. The precipitate was filtered and washed with warm ethanol. The filtrate was concentrated and the residue was chromatographed on silica (20% methanol in methylene chloride) to give 170 mg of the title compound 26a (5′-R) as a colorless solid. Similarly, the following compound was prepared: 4-amino-5-cyano-7- (5-
Deoxy-β-D-arabinofuranosyl) pyrrolo [2,3-d] -4-pyrimidone to 4-amino-5-cyano-7- (β-D-arabinofuranosyl) pyrrolo [
2,3-d] -4-Pyrimidone-5-carboxamidoxime (60) was prepared.

4-amino-5-cyano-7- (5-deoxy-β-D-ribofuranosyl) pyrrolo [2,3-d] -4-pyrimidone to 4-amino-5-cyano-7- (5-
Deoxy-β-D-ribofuranosyl) pyrrolo [2,3-d] -4-pyrimidone-
5-carboxamidoxime (13) was prepared. 4-amino-5-cyano-7-
(2,3-Didehydro-2,3-dideoxy-β-D-ribofuranosyl) pyrrolo [2,3-d] -4-pyrimidone to 4-amino-5-cyano-7- (2,3-
Didehydro-2,3-dideoxy-β-D-ribofuranosyl) pyrrolo [2,3-
d] -4-Pyrimidone-5-carboxamidoxime (51) was prepared. 4-Amino-5-cyano-7- (2-deoxy-β-D-ribofuranosyl) pyrrolo [2
Preparation of 4-amino-5-cyano-7- (2-deoxy-β-D-ribofuranosyl) pyrrolo [2,3-d] -4-pyrimidone-5-carboxamidoxime from 3,3-d] -4-pyrimidone did.

7- (5 (R) -C-methyl-β-D-ribofuranosyl) pyrrolo [2,3-d
] -4-Pyrimidone-5-carboxamidine hydrochloride (27a) Preparation 7- (5 (R) -C-methyl-β-D-ribofuranosyl) pyrrolo [2,3-d
] -4-Pyrimidone-5-carboxamidoxime (110 mg, 0.324 mm
ol), ammonium chloride (20 mg, 0.374 mmol) and Raney nickel (50% slurry in water, 200 mg) in water (75 mL) was shaken in a hydrogenator (H ₂ , 50 psi) at room temperature for 18 hours. . The catalyst was filtered and washed (warm water). The combined filtrate was concentrated and the product was recrystallized from methanol to give 100 mg of the title compound 27a (5′-R) as a colorless solid. The following compound was similarly prepared: 4-amino-5-cyano-7- (5-deoxy-β-D-
Arabinofuranosyl) pyrrolo [2,3-d] -4-pyrimidone-5-carboxamidoxime to 4-amino-5-cyano-7- (β-D-arabinofuranosyl)
Pyrrolo [2,3-d] -4-pyrimidone-5-carboxamidine hydrochloride (63) was prepared. 4-Amino-5-cyano-7- (5-deoxy-β-D-
Ribofuranosyl) pyrrolo [2,3-d] -4-pyrimidone-5-carboxamidoxime to 4-amino-5-cyano-7- (5-deoxy-β-D-ribofuranosyl) pyrrolo [2,3-d]- 4-Pyrimidone-5-carboxamidine hydrochloride (15) was prepared. 4-amino-5-cyano-7- (2-deoxy-β
-D-ribofuranosyl) pyrrolo [2,3-d] -4-pyrimidone-5-carboxamidoxime to 4-amino-5-cyano-7- (2-deoxy-β-D-ribofuranosyl) pyrrolo [2,3- d] -4-Pyrimidone-5-carboxamidine hydrochloride (70) was prepared.

As mentioned above, specific embodiments and applications of pyrrolo [2,3-d] pyrimidine nucleoside analogues have been disclosed. However, it will be apparent to one of ordinary skill in the art that many additional modifications other than those already disclosed are possible without departing from the inventive concept herein. The invention is, therefore, not limited except by the scope of the appended claims. Moreover, in interpreting the specification and claims, all terms should be interpreted in the broadest possible manner consistent with the context. In particular, "include (comp
The terms "rises" and "comprising" refer to elements, components or steps non-exclusively, and the elements, components or steps referred to are other elements, components not explicitly mentioned. Or it should be construed as indicating that it can be present with, used with, or combined with a step.

[Brief description of drawings]

[Figure 1]   1 is a first exemplary synthetic scheme of reactions involved in the preparation of compounds according to the present invention.

[Fig. 2]   2 is a second exemplary synthetic scheme of reactions involved in the preparation of compounds according to the present invention.

[Figure 3]   3 is a third exemplary synthetic scheme of reactions involved in the preparation of compounds according to the present invention.

[Figure 4]   4 is a fourth exemplary synthetic scheme for the reactions involved in the preparation of compounds according to the present invention.

[Figure 5]   5 is a fifth exemplary synthetic scheme of reactions involved in the preparation of compounds according to the present invention.

[Figure 6]   6 is a sixth exemplary synthetic scheme of reactions involved in the preparation of compounds according to the present invention.

[Figure 7]   1 illustrates exemplary compounds according to the present invention.

FIG. 8A is a graph showing the effect of each compound according to the present invention on the expression of type 1 cytokine.

FIG. 8B is a graph showing the effect of each compound according to the present invention on the expression of type 2 cytokine.

[Figure 9]   3 is a table showing the cytotoxicity of various compounds according to the present invention.

FIG. 10 is a table showing rates of DNA synthesis in cells treated with various compounds according to the present invention.

FIG. 11 is a graph showing inhibition of VEGF release from human prostate cancer cells when treated with compounds according to the present invention.

FIG. 12 is a graph showing inhibition of IL-8 release from human prostate cancer cells when treated with compounds according to the present invention.

[Procedure Amendment] Patent Cooperation Treaty Article 19 Amendment Translation Form

[Submission date] January 19, 2001 (2001.1.19)

[Procedure Amendment 1]

[Document name to be amended] Statement

[Name of item to be amended] Claims

[Correction method] Change

[Contents of correction]

[Claims]

[Chemical 1] [Wherein A is O, S or CH ₂ , X is H, NH ₂ or OH, and Y is H
, Halogen or NH ₂ and Z is H, halogen, R, OH, OR, SH, SR, NH ₂ , NHR, NR ₂ , CN, C (O) NH ₂ , COOH, COOR, CH ₂ NH _2. , C (= NOH)
NH _2, and C (= NH) is selected from the group consisting of NH _2, R is alkyl, alkenyl, alkynyl or aralkyl, _{R 2} and _{R 3} are independently from the group consisting of H, F and OH R ₄ is selected from the group consisting of hydrogen, alkyl, alkenyl, alkynyl and aralkyl, R ₄ optionally has at least one of a heteroatom and a functional group, and R ₅ is OH, OP (O) (OH) ₂ , P (O) (OH) ₂ , OP (O) (O
R ′) ₂ or P (O) (OR ′) ₂ , R ′ is a masking group, and R _{5 ′} is selected from the group consisting of alkyl, alkenyl, alkynyl and aralkyl, and R _{5 ′} is Having at least 2 carbon atoms and optionally having at least one of a heteroatom and a functional group].

[Chemical 2] [In the formula, Z is CN, C (O) NH ₂ , C (= NH) NH ₂ or C (= NOH) N
H ₂ and R ₄ and R _{5 ′} are independently selected from the group consisting of hydrogen, alkyl, alkenyl, alkynyl and aralkyl, R ₄ and R _{5 ′} are independently and optionally a heteroatom and A nucleoside analogue according to claim 1, comprising at least one of the functional groups, provided that R ₄ and R _{5 ′} are not both hydrogen.

[Chemical 3] [Wherein A is O, S or CH ₂ , X is H, NH ₂ or OH, and Y is H
, Halogen or NH ₂ and Z is H, halogen, R, OH, OR, SH, SR, NH ₂ , NHR, NR ₂ , CN, C (O) NH ₂ , COOH, COOR, CH ₂ NH _2. , C (= NOH)
NH _2, and C (= NH) is selected from the group consisting of NH _2, R is alkyl, alkenyl, alkynyl or aralkyl, _{R 2} and _{R 3} are independently from the group consisting of H, F and OH R ₄ and R _{5 ′} are selected from the group consisting of hydrogen, alkyl, alkenyl, alkynyl and aralkyl, R ₄ and R _{5 ′} are independently and optionally of a heteroatom and a functional group. At least one, R ₅ is H, OH, OP (O) (OH) ₂ , P (O) (OH) ₂ , OP (O)
(OR ') ₂ or P (O) (OR') ₂ and R'is a masking group], and to a concentration of the compound effective to alter the secretion of cytokines. A method of altering secretion of a cytokine from a cell, which comprises contacting the cell.

[Chemical 4] [In the formula, Z is CN, C (O) NH ₂ , C (= NH) NH ₂ , C (= NNH ₂ ) N
An H ₂ or _{-C (= NOH) NH 2,} R 5 _{is, H, OH, OP (O} ) (OH) 2, P (O) (OH) 2, OP (O)
(OR ') ₂ or P (O) (OR') ₂ and R'is a masking group], and to a concentration of the compound effective to alter the secretion of cytokines. A method of altering secretion of a cytokine from a cell, which comprises contacting the cell.

[Chemical 5] [Wherein A is O, S or CH ₂ , X is H, NH ₂ or OH, and Y is H
, Halogen or NH ₂ and Z is H, halogen, R, OH, OR, SH, SR, NH ₂ , NHR, NR ₂ , CN, C (O) NH ₂ , COOH, COOR, CH ₂ NH _2. , C (= NOH)
NH _2, and C (= NH) is selected from the group consisting of NH _2, R is alkyl, alkenyl, alkynyl or aralkyl, _{R 2} and _{R 3} are independently from the group consisting of H, F and OH R ₄ is selected from the group consisting of hydrogen, alkyl, alkenyl, alkynyl and aralkyl, R ₄ optionally comprises at least one of a heteroatom and a functional group, R _{5 ′} is hydrogen, Selected from the group consisting of alkyl, alkenyl, alkynyl and aralkyl, R _{5 ′} has at least 2 carbon atoms and optionally at least one of a heteroatom and a functional group, R ₅ is H, OH, OP (O) (OH) ₂ , P (O) (OH) ₂ , OP (O)
(OR ′) ₂ or P (O) (OR ′) ₂ and R ′ is a masking group,
Provided that both R ₄ and R _{5 ′} are not hydrogen], and the hyperproliferative cells are treated with a concentration of the compound effective to inhibit proliferation of the hyperproliferative cells. A method of suppressing the proliferation of hyperproliferative cells, comprising contacting.

20. The method of claim 19, wherein the cancer cells are prostate cancer cells.

[Procedure for Amendment] Submission for translation of Article 34 Amendment of Patent Cooperation Treaty

[Submission date] March 26, 2001 (2001.3.26)

[Procedure Amendment 1]

[Document name to be amended] Statement

[Name of item to be amended] Claims

[Correction method] Change

[Contents of correction]

[Claims]

[Chemical 1] [Wherein A is O, S or CH ₂ , X is H, NH ₂ or OH, and Y is H
, Halogen or NH ₂ and Z is H, halogen, R, OH, OR, SH, SR, NH ₂ , NHR, NR ₂ , CN, C (O) NH ₂ , COOH, COOR, CH ₂ NH _2. , C (= NOH)
NH _2, and C (= NH) is selected from the group consisting of NH _2, R is alkyl, alkenyl, alkynyl or aralkyl, _{R 2} and _{R 3} are independently from the group consisting of H, F and OH R ₄ is selected from the group consisting of hydrogen, alkyl, alkenyl, alkynyl and aralkyl, R ₄ optionally has at least one of a heteroatom and a functional group, and R ₅ is OH, OP (O) (OH) ₂ , P (O) (OH) ₂ , OP (O) (O
R ′) ₂ or P (O) (OR ′) ₂ , R ′ is a masking group, and R _{5 ′} is selected from the group consisting of alkyl, alkenyl, alkynyl and aralkyl, and R _{5 ′} is Having at least 2 carbon atoms and optionally having at least one of a heteroatom and a functional group].

[Chemical 2] [In the formula, Z is CN, C (O) NH ₂ , C (= NH) NH ₂ or C (= NOH) N
H ₂ and R ₄ and R _{5 ′} are independently selected from the group consisting of hydrogen, alkyl, alkenyl, alkynyl and aralkyl, R ₄ and R _{5 ′} are independently and optionally a heteroatom and A nucleoside analogue according to claim 1, comprising at least one of the functional groups, provided that R ₄ and R _{5 ′} are not both hydrogen.

[Chemical 3] [Wherein A is O, S or CH ₂ , X is H, NH ₂ or OH, and Y is H
, Halogen or NH ₂ and Z is H, halogen, R, OH, OR, SH, SR, NH ₂ , NHR, NR ₂ , CN, C (O) NH ₂ , COOH, COOR, CH ₂ NH _2. , C (= NOH)
NH _2, and C (= NH) is selected from the group consisting of NH _2, R is alkyl, alkenyl, alkynyl or aralkyl, _{R 2} and _{R 3} are independently from the group consisting of H, F and OH R ₄ and R _{5 ′} are selected from the group consisting of hydrogen, alkyl, alkenyl, alkynyl and aralkyl, R ₄ and R _{5 ′} are independently and optionally of a heteroatom and a functional group. At least one, R ₅ is H, OH, OP (O) (OH) ₂ , P (O) (OH) ₂ , OP (O)
(OR ') ₂ or P (O) (OR') ₂ and R'is a masking group], and to a concentration of the compound effective to alter the secretion of cytokines. A method of altering secretion of a cytokine from a cell, which comprises contacting the cell.

[Chemical 4] [In the formula, Z is CN, C (O) NH ₂ , C (= NH) NH ₂ , C (= NNH ₂ ) N
An H ₂ or _{-C (= NOH) NH 2,} R 5 _{is, H, OH, OP (O} ) (OH) 2, P (O) (OH) 2, OP (O)
(OR ') ₂ or P (O) (OR') ₂ and R'is a masking group], and to a concentration of the compound effective to alter the secretion of cytokines. A method of altering secretion of a cytokine from a cell, which comprises contacting the cell.

[Chemical 5] [Wherein A is O, S or CH ₂ , X is H, NH ₂ or OH, and Y is H
, Halogen or NH ₂ and Z is H, halogen, R, OH, OR, SH, SR, NH ₂ , NHR, NR ₂ , CN, C (O) NH ₂ , COOH, COOR, CH ₂ NH _2. , C (= NOH)
NH _2, and C (= NH) is selected from the group consisting of NH _2, R is alkyl, alkenyl, alkynyl or aralkyl, _{R 2} and _{R 3} are independently from the group consisting of H, F and OH R ₄ is selected from the group consisting of hydrogen, alkyl, alkenyl, alkynyl and aralkyl, R ₄ optionally comprises at least one of a heteroatom and a functional group, R _{5 ′} is hydrogen, Selected from the group consisting of alkyl, alkenyl, alkynyl and aralkyl, R _{5 ′} has at least 2 carbon atoms and optionally at least one of a heteroatom and a functional group, R ₅ is H, OH, OP (O) (OH) ₂ , P (O) (OH) ₂ , OP (O)
(OR ′) ₂ or P (O) (OR ′) ₂ and R ′ is a masking group,
Provided that both R ₄ and R _{5 ′} are not hydrogen], and the hyperproliferative cells are treated with a concentration of the compound effective to inhibit proliferation of the hyperproliferative cells. A method of suppressing the proliferation of hyperproliferative cells, comprising contacting.

[Procedure Amendment 2]

[Document name to be amended] Statement

[Correction target item name] 0008

[Correction method] Change

[Contents of correction]

[0008]

[Chemical 6] [Wherein A is O, S or CH_TwoAnd X is H, NH_TwoOr OH and Y is H
, Halogen or NH_TwoAnd Z is H, halogen, R, OH, OR, SH, S
R, NH_Two, NHR, NR_Two, CN, C (O) NH_Two, COOH, COOR, C
H_TwoNH_Two, C (= NOH) NH_Two, And C (= NH) NH_TwoSelected from the group consisting of
R is alkyl, alkenyl, alkynyl or aralkyl, R is_Two And R_ThreeAre independently selected from the group consisting of H, F and OH, R_FourIs hydrogen,
R is selected from the group consisting of alkyl, alkenyl, alkynyl and aralkyl, R _Four Optionally has at least one of a heteroatom and a functional group, R₅Is H,
OH, OP (O) (OH)_Two, P (O) (OH)_Two, OP (O) (OR ')_Twoor
Is P (O) (OR ')_Two, R'is a masking group, and R_{5 '}Is
Selected from the group consisting of alkyl, alkenyl, alkynyl and aralkyl,
R_{5 '}Has at least 2 carbon atoms, and optionally heteroatoms and functional groups
Have at least one of A pyrrolo [2,3-d] pyrimidine nucleoside analogue having a structure according to
is there.

[Procedure 3]

[Document name to be amended] Statement

[Correction target item name] 0010

[Correction method] Change

[Contents of correction]

[0010]

[Chemical 7] [In the formula, Z is CN, C (O) NH ₂ , C (= NH) NH ₂ or C (= NOH) N
H ₂ and R ₄ and R _{5 ′} are independently selected from the group consisting of hydrogen, alkyl, alkenyl, alkynyl and aralkyl, R ₄ and R _{5 ′} are independently and optionally a heteroatom and A pyrrolo [2,3-d] pyrimidine nucleoside having a structure according to [1], which contains at least one of the functional groups, provided that R ₄ and R _{5 ′} are not both hydrogen.

[Procedure amendment 4]

[Document name to be amended] Statement

[Name of item to be corrected] 0015

[Correction method] Change

[Contents of correction]

[0015]

[Chemical 8] [Wherein A is O, S or CH_TwoAnd X is H, NH_TwoOr OH and Y is H
, Halogen or NH_TwoAnd Z is H, halogen, R, OH, OR, SH, S
R, NH_Two, NHR, NR_Two, CN, C (O) NH_Two, COOH, COOR, C
H_TwoNH_Two, C (= NOH) NH_Two, And C (= NH) NH_TwoSelected from the group consisting of
R is alkyl, alkenyl, alkynyl or aralkyl, R is_Two And R_ThreeAre independently selected from the group consisting of H, F and OH, R_FourIs hydrogen,
R is selected from the group consisting of alkyl, alkenyl, alkynyl and aralkyl, R _Four Optionally has at least one of a heteroatom and a functional group, R₅Is H,
OH, OP (O) (OH)_Two, P (O) (OH)_Two, OP (O) (OR ')_Twoor
Is P (O) (OR ')_Two, R'is a masking group, and R_{5 '}Is
Selected from the group consisting of alkyl, alkenyl, alkynyl and aralkyl,
R_{5 '}Has at least 2 carbon atoms, and optionally heteroatoms and functional groups
Have at least one of. ]   As used herein, "alkyl", "alkenyl", "alkynyl" and "a
It should be especially appreciated that the term "rualkyl" refers to both straight and branched types.
Substituent R_TwoAnd R_ThreeRegarding R_TwoAnd R_ThreeBoth independently pair α or β planes.
You should be aware that you can be an elephant. Furthermore, C_{5 '}The above substitutions are non-identical
C₅The above substitutions may result in R or S chiral centers. When used here
The term "heteroatom" refers to atoms other than carbon in organic molecules and is
Terror atoms include halogen, nitrogen, oxygen and sulfur. When used here, "sensual
The term “group” refers to a reactive bond (eg, a double or triple bond) or a reactive group (eg, —OH).
, -SH, -NH_Two, -N_Three, -CN, -COOH, -CHO, -CONH_Two,
And so on).

[Procedure Amendment 5]

[Document name to be amended] Statement

[Correction target item name] 0018

[Correction method] Change

[Contents of correction]

[0018]

[Chemical 9] [In the formula, Z is CN, C (O) NH ₂ , C (= NH) NH ₂ or C (= NOH) N
H ₂ and R ₄ and R _{5 ′} are independently selected from the group consisting of hydrogen, alkyl, alkenyl, alkynyl and aralkyl, R ₄ and R _{5 ′} are independently and optionally a heteroatom and Containing at least one of the functional groups, provided that R ₄ and R _{5 ′} are not both hydrogen, and the remaining substituents are as defined in formula (I).

[Procedure correction 6]

[Document name to be amended] Statement

[Name of item to be corrected] 0027

[Correction method] Change

[Contents of correction]

[0027]

[Chemical 10] [In the formula, Z is CN, C (O) NH ₂ , C (= NH) NH ₂ , C (= NNH ₂ ) N
H ₂ or —C (= NOH) NH ₂ , and R ₅ is H, OH, OP (O) (OH
) ₂ , P (O) (OH) ₂ , OP (O) (OR ') ₂ or P (O) (OR') ₂ and R'is a masking group].

[Procedure Amendment 7]

[Document name to be corrected] Drawing

[Name of item to be corrected] Figure 1

[Correction method] Change

[Contents of correction]

[Figure 1]

[Procedure Amendment 8]

[Document name to be corrected] Drawing

[Name of item to be corrected] Figure 2

[Correction method] Change

[Contents of correction]

[Fig. 2]

[Procedure Amendment 9]

[Document name to be corrected] Drawing

[Name of item to be corrected] Figure 3

[Correction method] Change

[Contents of correction]

[Figure 3]

[Procedure Amendment 10]

[Document name to be corrected] Drawing

[Name of item to be corrected] Fig. 4

[Correction method] Change

[Contents of correction]

[Figure 4]

[Procedure Amendment 11]

[Document name to be corrected] Drawing

[Name of item to be corrected] Figure 5

[Correction method] Change

[Contents of correction]

[Figure 5]

[Procedure Amendment 12]

[Document name to be corrected] Drawing

[Name of item to be corrected] Figure 6

[Correction method] Change

[Contents of correction]

[Figure 6]

[Procedure Amendment 13]

[Document name to be corrected] Drawing

[Name of item to be corrected] Figure 7

[Correction method] Change

[Contents of correction]

[Figure 7]

─────────────────────────────────────────────────── ─── Continuation of front page (51) Int.Cl. ⁷ Identification code FI theme code (reference) A61P 35/00 A61P 35/00 37/06 37/06 43/00 105 43/00 105 C07H 19/14 C07H 19/14 (81) Designated countries EP (AT, BE, CH, CY, DE, DK, ES, FI, FR, GB, GR, IE, IT, LU, MC, NL, PT, SE), OA ( BF, BJ, CF, CG, CI, CM, GA, GN, GW, ML, MR, NE, SN, TD, TG), AP (GH, GM, KE, LS, MW, MZ, SD, SL, SZ, TZ, UG, ZW), EA (AM, AZ, BY, KG, KZ, MD, RU, TJ, TM), AE, AG, AL, AM, AT, AU, AZ, BA, BB, B G, BR, BY, BZ, CA, CH, CN, CR, CU, CZ, DE, DK, DM, DZ, EE, ES, FI, GB, GD, GE, GH, GM, HR, HU, ID , IL, IN, IS, JP, KE, KG, KP, KR, KZ, LC, LK, LR, LS, LT, LU, LV, MA, MD, MG, MK, MN, MW, MX, MZ, NO, NZ, PL, PT, RO, RU, SD, SE, SG, SI, SK, SL, TJ, TM, TR, TT, TZ, UA, UG, US, UZ, VN, YU, ZA, ZW (72) Inventor Tam, Robert United States, California 92626, Costa Mesa, Highland Avenyu 3300, ICN Pharmacy Youthicals Incorporated (72) Inventor Pietrskovsky, Zbigniew America United States, California, 92626, Costa Mesa, Highland Avenyu 3300, ICN Farmayu Youthals Incorporated F-term (reference) 4C050 AA01 BB04 CC08 EE03 FF01 GG03 GG04 GG07 HH04 4C057 A0317A02 LL20 LL26 4C086 AA01 AA02 AA03 CB05 EA06 GA02 GA07 MA01 MA04 MA17 MA23 MA34 MA35 MA52 MA56 MA63 MA66 NA14 ZB08 ZB11 ZB21 ZB26 ZB33 [Continued Summary]

Claims (20)

[Claims] 1. Formula (I): [Wherein A is O, S or CH ₂ , X is H, NH ₂ or OH, and Y is H
, Halogen or NH ₂ and Z is H, halogen, R, OH, OR, SH, SR, NH ₂ , NHR, NR ₂ , CN, C (O) NH ₂ , COOH, COOR, CH ₂ NH _2. , C (= NOH)
NH _2, and C (= NH) is selected from the group consisting of NH _2, R is alkyl, alkenyl, alkynyl or aralkyl, _{R 2} and _{R 3} are independently from the group consisting of H, F and OH R ₄ is selected from the group consisting of hydrogen, alkyl, alkenyl, alkynyl and aralkyl, R ₄ optionally has at least one of a heteroatom and a functional group, and R ₅ is H, OH , OP (O) (OH) ₂ , P (O) (OH) ₂ , OP (O)
(OR ′) ₂ or P (O) (OR ′) ₂ , R ′ is a masking group, and R _{5 ′} is selected from the group consisting of alkyl, alkenyl, alkynyl and aralkyl, R _{5 ′} Has at least 2 carbon atoms and optionally has at least one of a heteroatom and a functional group]. 2. Z is CN, C (O) NH ₂ or C (═NH) NH ₂ ,
The nucleoside analog according to claim 1, wherein R _{5 ′} has at least 2 carbon atoms and is selected from the group consisting of alkyl, alkenyl, alkynyl and aralkyl. 3. Structure: embedded image [In the formula, Z is CN, C (O) NH ₂ , C (= NH) NH ₂ or C (= NOH) N
H ₂ and R ₄ and R _{5 ′} are independently selected from the group consisting of hydrogen, alkyl, alkenyl, alkynyl and aralkyl, R ₄ and R _{5 ′} are independently and optionally a heteroatom and A nucleoside analogue according to claim 1, comprising at least one of the functional groups, provided that R ₄ and R _{5 ′} are not both hydrogen. 4. Formula (II): embedded image [Wherein A is O, S or CH ₂ , X is H, NH ₂ or OH, and Y is H
, Halogen or NH ₂ and Z is H, halogen, R, OH, OR, SH, SR, NH ₂ , NHR, NR ₂ , CN, C (O) NH ₂ , COOH, COOR, CH ₂ NH _2. , C (= NOH)
NH _2, and C (= NH) is selected from the group consisting of NH _2, R is alkyl, alkenyl, alkynyl or aralkyl, _{R 2} and _{R 3} are independently from the group consisting of H, F and OH R ₄ and R _{5 ′} are selected from the group consisting of hydrogen, alkyl, alkenyl, alkynyl and aralkyl, R ₄ and R _{5 ′} are independently and optionally of a heteroatom and a functional group. At least one, R ₅ is H, OH, OP (O) (OH) ₂ , P (O) (OH) ₂ , OP (O)
(OR ') ₂ or P (O) (OR') ₂ and R'is a masking group], and to a concentration of the compound effective to alter the secretion of cytokines. A method of altering secretion of a cytokine from a cell, which comprises contacting the cell. 5. The method of claim 4, wherein the cytokine is a type 1 cytokine. 6. The method of claim 5, wherein the type 1 cytokine is IFNγ. 7. The method of claim 4, wherein the cytokine is a type 2 cytokine. 8. The method of claim 7, wherein the type 2 cytokine is IL-4. 9. The method of claim 4, wherein the cells are lymphocytes. 10. The method of claim 4, wherein the cells are cancer cells. 11. The method of claim 10, wherein the cancer cells are prostate cancer cells. 12. Formula (III): embedded image [In the formula, Z is CN, C (O) NH ₂ , C (= NH) NH ₂ , C (= NNH ₂ ) N
An H ₂ or _{-C (= NOH) NH 2,} R 5 _{is, H, OH, OP (O} ) (OH) 2, P (O) (OH) 2, OP (O)
(OR ') ₂ or P (O) (OR') ₂ and R'is a masking group], and to a concentration of the compound effective to alter the secretion of cytokines. A method of altering secretion of a cytokine from a cell, which comprises contacting the cell. 13. Formula (IV): embedded image [Wherein A is O, S or CH ₂ , X is H, NH ₂ or OH, and Y is H
, Halogen or NH ₂ and Z is H, halogen, R, OH, OR, SH, SR, NH ₂ , NHR, NR ₂ , CN, C (O) NH ₂ , COOH, COOR, CH ₂ NH _2. , C (= NOH)
NH _2, and C (= NH) is selected from the group consisting of NH _2, R is alkyl, alkenyl, alkynyl or aralkyl, _{R 2} and _{R 3} are independently from the group consisting of H, F and OH R ₄ is selected from the group consisting of hydrogen, alkyl, alkenyl, alkynyl and aralkyl, R ₄ optionally comprises at least one of a heteroatom and a functional group, R _{5 ′} is hydrogen, Selected from the group consisting of alkyl, alkenyl, alkynyl and aralkyl, R _{5 ′} has at least 2 carbon atoms and optionally at least one of a heteroatom and a functional group, R ₅ is H, OH, OP (O) (OH) ₂ , P (O) (OH) ₂ , OP (O)
(OR ′) ₂ or P (O) (OR ′) ₂ and R ′ is a masking group,
Provided that both R ₄ and R _{5 ′} are not hydrogen], and the hyperproliferative cells are treated with a concentration of the compound effective to inhibit proliferation of the hyperproliferative cells. A method of suppressing the proliferation of hyperproliferative cells, comprising contacting. 14. The method of claim 13, wherein the hyperproliferative cells are cancer cells. 15. The method of claim 14, wherein the cancer cells are prostate cancer cells. 16. The method of claim 13, wherein inhibiting growth comprises inhibiting DNA synthesis. 17. Release of growth factor from a cell comprising providing a compound according to claim 1 and contacting said cell with a concentration of said compound effective to inhibit release of growth factor. How to suppress. 18. The method of claim 17, wherein the growth factor is VEGF. 19. The method of claim 17, wherein the cell is a cancer cell. 20. The method of claim 19, wherein the cancer cells are prostate cancer cells.