HRP980477A2 - Cytokine related treatments of disease - Google Patents

Description

Field of invention

The invention is from the field of pharmacology and biochemistry, more precisely from the field of nucleosides.

State of the art

The cells of the mammalian immune system contain two main classes of lymphocytes: B lymphocytes (B cells), which are produced in the bone marrow, and T lymphocytes (T cells), which are produced in the thymus. B cells are mainly responsible for humoral immunity (ie antibody production), while T cells are responsible for cell-directed immunity.

T cells are generally considered to fall into two subclasses, helper T cells and cytotoxic T cells. Helper T cells activate other lymphocytes, including B cells and cytotoxic T cells and macrophages, by releasing soluble protein mediators called cytokines that are involved in cell-directed immunity. As used herein, lymphokines are a subset of cytokines.

T cells are generally considered to fall into two subtypes, Th1 and Th2. Th1 cells (also known as type 1 cells) produce interleukin 2 (IL-2), tumor necrosis factor (TNFa) and gamma interferon (IFNy), and are responsible for primary, cell-directed immunity, such as delayed-type hypersensitivity and against -viral immunity. In contrast, Th2 cells (also known as type 2 cells) produce interleukins, IL-4, IL-5, IL-6, IL-9, IL-10, and IL-13 and are primarily involved in assisting humoral immune responses as what are those to allergens, eg, IgE and IgG4 antibody isotype overlap (Mosmann, 1989, Annu Rev Immunol, 7:145-173).

As used herein, Thl and Th2 "responses" mean that the full range of effects elicited by Th1 and Th2 lymphocytes, respectively, is involved. Among other things, such responses include changes in the production of appropriate cytokines through transcription, translation, secretion, and possibly other mechanisms, increased proliferation of appropriate lymphocytes, and other effects associated with increased cytokine production, including motility effects.

The prior applications, which are incorporated herein by reference, relate to aspects of our more recent discoveries including the action of various nucleosides (as defined herein, including derivatives and analogs of natural nucleosides) to selectively modulate lymphocyte responses relative to one another. Among other things, we have shown that either Th1 or Th2 responses can be selectively suppressed, while the other can either be induced or remain relatively unchanged. We also surprisingly discovered that nucleosides that are effective in selectively modulating Th1 and Th2 responses relative to each other tend to exhibit a dual effect. Among other things, nucleosides that tend to generally suppress or induce both Th1 and Th2 activity at relatively higher doses tend to selectively modulate Th1 and Th2 relative to each other at relatively lower doses.

The mechanisms by which nucleosides and other compounds selectively modulate Th1 and Th2 responses relative to each other are still unclear. One postulated by the inventors of this invention is that effective nucleosides alter the total amount of guanisine triphosphate (GTP), which in turn affects the rate at which cytokines are produced. According to this theory, relatively large variations in available GTP are sufficient to affect the concentration of Th1 and Th2 cytokines, while relatively smaller variations in available GTP tend to alter the concentration of Th1 and Th2 cytokines with different reach.

The effects of 2-ß-D-ribofuranosylthiazole-4-carboxamide (Tiazofurin), a moderate C-nucleoside analog, at the GTP level support this. Tumor cells are characterized by a high level of inosine monophosphate dehydrogenase (IMP DH) activity, and it is known that IMP DH is the enzyme that determines the overall rate of GTP biosynthesis. Weber, G., IMP Dehydrogenase and GTP as Targets in Human Leukemia Treatment, Adv. Exp. Honey. Biol. 309B:287-292 (1991). It has been shown that thiazofuran selectively stops the activity of IMP DH and reduces the total amount of guanine nucleotide, which in turn promotes remission of various tumors. Weber, G., Critical Issues in Chemotherapy with Tiazofurin, Adv. Enzyme Regul. 29:75-95 (1989). Usual initial doses of Thiazofuran are about 4,400 mg/m2, with consolidating doses of about 1,100 to 3,300 mg/m2. At these levels of synthesis, Th1 and Th2 responses are greatly reduced, thereby significantly lowering the level of the immune system. In one aspect of the present invention, much lower doses of Thiazofurin, ranging from 1/10 to 1/2 above, are believed to be sufficient to specifically suppress either a Th1 or Th2 response, without significantly reducing the other response.

The action of 1-ß-D-ribofuranosyl-1,2,4-triazole-3-carboxamide (Ribavirin) also supports this theory. Ribavirin is a potent broad-spectrum antiviral agent that has been shown to inhibit IMP DH. Yamada, Y. et al., Action of the Active Metabolites of Tiazofurin and Ribavirin on Purified IMP Dehydrogenase, Biochem. 27:2193-2196 (1988). When inhibiting IMP DH. Ribavirin works by a different mechanism than Thiazofuran, however, it works at a different place on the enzyme molecule. Ribavirin is converted to its active metabolite, ribavirin monophosphate (RMP), which inhibits the enzyme at the IMP-XMP site of the IMP DH. As with Tiazofurin, the affinity of the active form of Ribavirin for the enzyme is higher than for the natural metabolite. At relatively high doses, approximately 2200 mg/m2 or about 1200-1500 mg/day for an adult, Ribavirin reduces the activity of IMP DH so much that Th1 and Th2 responses are inhibited. At lower doses, approximately 600 to 1000 mg/day, Ribavirin promotes a Th1 response and suppresses a Th2 response.

Regardless of the existence of mechanisms that have not yet been defined, we found that a potentially huge positive effect can be achieved by modulating Th1 and Th2 responses with each other. We have concluded, for example, that specific modulation of Th1 relative to Th2 may be beneficial in the treatment of various conditions and diseases, ranging from infections, infestations, tumors and hypersensitivity to autoimmune diseases.

These findings are particularly significant because current treatment strategies for many of the above-mentioned diseases have either limited efficacy, significant side effects, or both. Treatment of autoimmune diseases, for example, is often limited to palliative procedures, removal of toxic antibodies (as in myasthenia gravis), use of aggressive drugs including corticosteroids, chloroquine derivatives, antimetabolic or antitumor drugs, and drugs such as ciclosporin that affect cells of the immune system.

SUMMARY OF THE INVENTION

This application refers to the use of nucleosides in a relatively small dose range with the aim of selectively modulating Th1 and Th2 responses relative to each other in the treatment of disease. In one aspect of the invention, administration of the nucleoside or other compound reduces the dose at which the primary drug is administered. In another aspect of the present invention, an abnormality expressed by an increased response in one group of cytokines is treated with the administration of a nucleoside or other compound that increases the response in another group of cytokines. In a further aspect of this invention, the patient is treated prophylactically with the use of a nucleoside or other compound that selectively reduces Th1 activity without significantly reducing Th2 activity. In a further aspect of the present invention, the nucleoside or other compound is administered to the patient at a dose that reduces the total amount of GTP to a level that selectively reduces one of the Th1 or Th2 responses without significantly reducing the other response. Forms with controlled release are especially intended to achieve such an effect.

Examples of nucleosides believed to act in this way are the D- and L-forms: (a) bicyclic nucleosides corresponding to any of Formula 1, 1-A to 1-F; and (b) monocyclic nucleosides corresponding to formulas 2 to 5.

Examples of primary drugs that are thought to work in this way are antiviral agents such as Ribavirin, acyclovir, and AZTTM; antifungal agents such as Tolnaftate, FungizoneTM, LotriminTM, MycelexTM, Nystatin and Amphoteracin; anti-parasitic agents such as MintezolTM, NiclocideTM, VermoxTM and FlagylTM; intestinal agents such as ImmodiumTM, LomotilTM and PhazymeTM; antitumor agents such as AdriamycinTM, CytoxanTM, ImuranTM, Methotrexate, MithracinTM, ThiazofurinTM, TaxolTM; dermatological agents such as AclovateTM, CyclocortTM, DenorexTM, FloroneTM, OxsoralenTM, activated charcoal and salicylic acid; anti-migraine preparations such as ergotamine compounds; steroids and immunosuppressants not listed above, including cyclosporine, DiprosoneTM, hydrocortisone; FloronTM, LidexTM, Topicort and Valisone; and metabolic agents such as insulin.

Exhausted description

Definitions

In this specification, the following terms are used, with the meanings given below.

The term "α" and "ί" indicate the specific stereochemical distribution of the substituents on the asymmetric carbon atom in the drawn chemical structure.

The term "abnormality" refers to a condition associated with a disease. Therefore, Th1 and/or Th2 responses resulting from autoimmune disease are considered here as an abnormality of the appropriate cytokine(s), although such cytokine responses may be a normal result of the disease.

The term "aryl" refers to monovalent unsaturated aromatic radicals having one ring (eg, phenyl) or two fused rings (eg, naphthyl), which may be optionally substituted with hydroxyl, lower alkyl, chlorine, and/or cyano.

The term "effective amount" refers to that amount of a compound of formula (I) which will bring the immune function to a normal level, or will increase the immune function above the normal level to clear the infection.

The term "enantiomers" refers to a pair of stereoisomers that are not overlapping mirror images. A mixture of enantiomeric pairs in a 1:1 ratio is a "racemic" mixture.

The term "heterocyclic compound" refers to monovalent saturated or unsaturated radicals containing at least one heteroatom, such as N, O or S, within the ring, while any available position can be arbitrarily substituted, independently, with hydroxy, oxo, amino groups , imino, lower alkyl, bromine, chlorine and/or cyano group. Within this class of substituents are purines and pyrimidines.

The term "immunological modulators" refers to natural and artificial products that can alter the normal or abnormal immune system by stimulating or suppressing it.

The term "isomers" refers to mutually different compounds that have the same formula.

"Stereoisomers" are isomers that differ from each other in the way the atoms are spatially distributed. The term "L-configuration" is used in this invention to describe the chemical configuration of ribofuranosyl for compounds which are bound to nucleic bases. The L-configuration of the sugar for the compounds of this invention is different from the D-configuration of the ribose sugar of natural nucleosides such as cytidine, adenosine, guanosine and uridine.

The term "lower alkyl" refers to methyl, n-propyl, isopropyl, n-butyl, t-butyl, i-butyl or n-hexyl. This term further refers to cyclic, branched or chain forms of one to six carbon atoms.

The term "monocyclic" refers to monovalent saturated carbon radicals having at least one heteroatom, such as O, N, S or P, within the ring, where each available site may be arbitrarily and independently substituted with a sugar or any other group such as are a bromine, chlorine and/or cyano group, so that the monocyclic system is optionally aromatized [e.g., thymidine; 1-(2'-deoxy-?-D-erythro-pentofuranosyl)thymine].

The term "nucleoside" refers to a compound composed of a pentose or a modified pentose that is attached at a specific site of a heterocyclic compound or at the natural site (position 9) of ourine or pyrimidine (position 1), or to an equivalent position of an analog, including in particular D- and L-forms of nitrogen bicyclic and monocyclic heterocyclic compounds shown in Figures 1, 1-A through 1-F, and Figures 2 through 5.

The term “C-nucleosides” is used throughout this specification to describe a bond formed between a ribose sugar and a heterocyclic base. In C-nucleosides, the bond starts from the C-1 position of the ribose and joins the carbon atom of the heterocyclic base. The bond formed in C-nucleosides is carbon-carbon type.

The term "D-nucleosides" refers to nucleoside compounds that have the sugar D-ribose (eg, adenosine).

The term "L-nucleosides" refers to nucleoside compounds having L-ribose.

The term “N-nucleosides” used in this specification describes the type of bond formed between the ribose sugar and the heterocyclic base. In N-nucleosides, the bond originates from the C-1 position of the ribose and joins the nitrogen atom of the heterocyclic base. The bond formed in C-nucleosides is of the carbon-nitrogen type.

The term "nucleotide" refers to the phosphate ester that is substituted in the 5'-position of the nucleoside.

The term "pharmaceutically acceptable salt" refers to any salt derived from inorganic and organic acids or bases.

The term "protecting group" refers to a chemical group that is attached to a carbon or nitrogen atom to prevent further reaction during the introduction of other species into the molecule containing the nitrogen or oxygen atom. Those familiar with the field of organic synthesis are familiar with the full range of oxygen and nitrogen protecting groups.

The term "purine" refers to the nitrogen bicyclic heterocyclic compounds shown in Figures 1, 1-A through 1-F.

The term "pyrimidine" refers to the nitrogen monocyclic heterocyclic compounds shown in Figures 2 through 5.

The term "tumor" refers broadly to any autonomous morbid tissue growth that may or may not be malignant, including neoplasms and carcinomas.

The term "treatment" or "treatment" of a disease refers to the implementation of a protocol, which may include the administration of one or more drugs to the patient, with the aim of avoiding the signs or symptoms of the disease. Thus, "treatment" or "treatment" does not require complete removal of signs or symptoms, does not require treatment, and specifically includes protocols that have only a superficial effect on the patient.

Combinations and methods

The contemplated combinations in one aspect of the present invention generally include a primary or “first” drug and a second or “secondary” drug, while the contemplated methods include selecting and combining the first and second drugs in combination therapy. In preferred embodiments, a disease is identified that is known to cause abnormalities of at least one cytokine in the patient, and the first drug is selected among the compounds that have shown an effect on the disease at a single therapeutic dosage, and the second drug, which can be the bimodal nucleoside modulator described here , which is selected from among those compounds known to aggravate the abnormalities produced by the disease, when administered in a given dose range. Then, the first drug was administered at a lower than monotherapeutic dose and the second drug at a dose outside the range of the dose that worsens the irregularity. Since the second drug shows bimodal activity against at least one cytokine of interest, the combination is nevertheless effective for the treatment of the disease, and the use of the second drug allows for a reduction in the dose of the primary or first drug.

Examples of primary drugs believed to be effective in combination with a modulator selected from those of Figure 1, 1A through 1F, and 2-5 are antiviral agents such as interferon, including but not limited to interferon α and γ, Ribavirin, acyclovir and AZTTM; antifungal agents such as Tolnaftate, FungizoneTM, LotriminTM MycelexTM, Nystatin and Amphoteracin; anti-parasitic agents such as MintezolTM, NiclocideTM, VermoxTM and FlagylTM; intestinal agents such as ImmodiumTM, LomotiTM and PhazymeTM; anti-tumor agents such as interferon α and γ, AdriamycinTM, CytoxanTM, ImuranTM, Methotrexate, MithracinTM, TiazofurinTM, TaxolTM; dermatological agents such as AclovateTM, CyclocortTM, DenorexTM, FloroneTM, OxsoralenTM, activated charcoal and salicylic acid; anti-migraine preparations such as ergotamine compounds; steroids and immunosuppressive agents not listed above, including cyclosporine, DiprosoneTM, hydrocortisone; FloronTM, LidexTM, Topicort and Valisone; and metabolic agents such as insulin, as well as other drugs that do not belong to the above categories, including cytokines such as IL2, IL4, IL6, IL8, IL10 and IL12. Particularly preferred drugs are AZT, 3TC, 8-substituted guanosine analogues, 2',3'-dideoxynucleosides, interleukin II, interferons such as IαB-interferons, tucaresol, levamisoles, isoprinosines and cyclolignan.

Examples of secondary drugs believed to be effective in the present invention are D- and L-forms (a) bicyclic nucleosides corresponding to the generic formula 1 from 1-A to 1-F, and (b) monocyclic nucleosides corresponding to formulas 2 to 5 Other nucleoside and non-nucleoside compounds that are effective in the present invention can be easily identified by screening such compounds in vitro for activity against IL-2, TNF-α, IFN-γ, IL-4 and IL-5, as described in PCT /US97/00600.

The compounds of formula 1 are purine nucleosides of the following structure:

[image]

where R1, R2, R3, R4, R5, R2' and R3' are independently selected from the group consisting of H, OH, NH2, F, Cl, Br, I, N3, -CN, -OR', -NR'2 , -SR', -NHNH2, -NHOH, CHO, COOR', CONR'2, alkyl, alkenyl, alkylnyl, aryl, aralkyl, substituted alkyl, substituted alkenyl, substituted alkynyl, substituted aryl, substituted aralkyl, wherein the substituent is selected from F, Cl, Br, I, N3, -CN, -OR", NO2, -NR"2, SR", -NHNH2, -NHOH, COOR", CONR"2 and where R' and R" are H, alkyl , alkenyl, alkynyl, aryl, aralkyl;

W = O, S, CH2, Se;

Z1 and Z2 are independently selected from N, C, CH;

Z3, Z4 and Z5 are independently selected from the group consisting of -CR-, -NR-, -O-, -S, -Se-, -C=O, -C=S, -S=O, -CR=CR -, -CR=N-, -N=N-, where R is selected from the group consisting of H, F, C1, Br, I, N3, -CN, -OR', -NR'2, -SR', -NHNH2, -NHOH, -NO2, CHO, COOR', CONH2, -C(O)-NH2, -C(S)-NH2, -C(NH)-NH2, -C(NOH)-NH2, =O , =NH, =NOH, =NR, alkyl, alkenyl, alkylnyl, aryl, aralkyl, substituted alkyl, substituted alkenyl, substituted alkynyl, substituted aryl, substituted aralkyl, where the substituent is selected from H, -OH, NH2, F, Cl , Br, I, N3, -CN, -COOR", -CONR"2, -OR", -NR"2, -SR", -NHNH2, -NHOH and -NO2, while R' and R" are H, alkyl, alkenyl, alkynyl, aryl, aralkyl, acetyl, acyl and sulfonyl;

The chemical bond between Z3 and Z4 or Z4 and Z5 is selected from C-C, C=C, C-N, C=N, N-N, N=N, C-S and N-S;

X and Y are independently selected from the group consisting of H, OH, NH2, F, Cl, Br, I, N3, -S-NH2, -S(O)-NH2, -S(O2)-NH2, -CN, -COOR', -CONR'2, -OR', -NR'2, -SR', NHNH2, -NHOH, alkyl, alkenyl, alkylnyl, aryl, aralkyl, substituted alkyl, substituted alkenyl, substituted alkynyl, substituted aryl, substituted aralkyl, where the substituent is selected from the group consisting of F, Cl, Br, I, N3, -CN, -OR", NO2, -NR"2, SR", -NHNH2, -NHOH while R', R" are H , alkyl, alkenyl, alkynyl, aryl, aralkyl.

Compounds of formula 1-A are 8-substituted "- or ß- L- or D- guanosine analogs of the following structure:

[image]

where X is selected from the group consisting of H, R, F, Cl, Br, I, N3, -CN, -OR, -SR, -NR2, -NHNH2, NHOH, -CHO, -CONH2, -COOR and -L-A ; where R is selected from the group consisting of alkyl, alkenyl, alkynyl and aralkyl, acetyl, acyl and sulfonyl; L is a linker selected from the group consisting of alkyl, alkenyl, alkynyl and aralkyl; while A is selected from the group consisting of H, -OR', -SR', -NR'2, -NHNR'2, -CHO, -COOR', -CONR'2, where R' is selected from H, Me, Et, allyl, acetyl and -COCF3;

Y is selected from H, R, F, Cl, Br, I, N3, CN, OR, SR and NR2, where R is selected from the group consisting of H, alkyl, alkenyl, alkynyl and aralkyl, acetyl, acyl, sulfonyl;

Z is N or CH; and

R1, R2 and R3 are independently selected from the group consisting of H, -OH, -OAc, -OBz, -OP(O2)OH.

Compounds of formula 1B are 7-substituted-8-oxo-" -or ß- L-guanosine analogues of the following structure:

[image]

Compounds of formula 1-C are 7-deaza-7,8-mono- or disubstituted " -or ß- L- or D-guanosine analogues of the following structure:

[image]

where X1 and X2 are independently selected from the group consisting of H, R, F, Cl, Br, I, N3, -CN, -OR, -SR, -NR2, -NHNH2, -NHOH, -CHO, -CONH2, - COOR and -L-A; where R is selected from the group consisting of alkyl, alkenyl, alkynyl and aralkyl, acetyl, acyl and sulfonyl; L is a linker selected from the group consisting of alkyl, alkenyl, alkynyl and aralkyl; and A is selected from the group consisting of H, -OR', -SR', -NR'2, -NHNR'2, -CHO, COOR', -CONR'2, where R' is selected from H, Me, Et , allyl, acetyl, -COCF3;

Y is selected from the group consisting of H, R, F, Cl, Br, I, N3, -CN, -OR, -SR, -NR2, where R is selected from the group consisting of H, alkyl, alkenyl, alkynyl and aralkyl , acetyl, acyl and sulfonyl;

Z is N or CH;

R1, R2 and R3 are independently selected from the group consisting of H, -OH, -OAc, -OBz, -OP(O2)OH.

Compounds of formula 1-D are 7-deaza-8-aza-7-substituted "-or ß- L- or D- guanosine analogs of the following structure:

[image]

where X is selected from the group consisting of H, R, F, Cl, Br, I, N3, -CN, -OR, -SR, -NR2, -NHNH2, NHOH, -CHO, -CONH2, -COOR and -L-A ; where R is selected from the group consisting of alkyl, alkenyl, alkynyl and aralkyl, acetyl, acyl and sulfonyl; L is a linker selected from the group consisting of alkyl, alkenyl, alkynyl and aralkyl; while A is selected from the group consisting of H, -OR', -SR', -NR'2, -NHNR'2, -CHO, -COOR', -CONR'2, where R' is selected from H, Me, Et, allyl, acetyl and -COCF3;

Y is selected from H, R, F, Cl, Br, I, N3, CN, OR, SR and NR2, where R is selected from the group consisting of H, alkyl, alkenyl, alkynyl and aralkyl, acetyl, acyl, sulfonyl;

Z is N or CH; and

R1, R2 and R3 are independently selected from the group consisting of H, -OH, -OAc, -OBz, -OP(O2)OH.

The compounds of formula 1-E are thiazolo[4,5-d]pyrimidine "-or ß- L- or D-nucleosides of the following structure:

[image]

X1 = O, S, =NH, =NNH2, =NHOH and =NR where R is selected from alkyl, alkyenyl, alkynyl and aralkyl, acyl;

X 2 is S, O or Se

Y is selected from the group consisting of H, R, F, Cl, Br, I, N3, -CN, -OR, -SR, -NR2, where R is selected from the group consisting of H, alkyl, alkenyl, alkynyl and aralkyl , acetyl, acyl and sulfonyl;

Z is N or CH;

R1, R2 and R3 are independently selected from the group consisting of H, -OH, -OAc, -OBz, -OP(O2)OH.

Compounds of formula 1-F are ß-L- or D- purine nucleosides of the following structure:

[image]

X is selected from H, R, -SNH2, -S(O)NH2, -SO2NH2, F, Cl, Br, I, N3, -CN, -OR, SR, -NR2, where R is selected from the group consisting of H, alkyl, alkenyl, alkynyl and aralkyl, acetyl, acyl and sulfonyl;

Y is selected from H, R, F, Cl, Br, I, N3, -CN, -OR, -SR, -NR2, where R is selected from the group consisting of H, alkyl, alkenyl, alkynyl and aralkyl, acetyl, acyl and sulfonyl;

Z 1 , Z 2 and Z 3 are independently selected from C, N and CH;

R1, R2 and R3 are independently selected from H, -OH, -OAc, -OBz and -OP(O2)OH.

Compounds of formula 2 have the following structures:

[image]

where:

A is independently selected from N or C;

B, C, E, F are independently selected from CH, CO, N, S, Se, O, NR1, CCONH2, CCH3, C-R2 or P; R 1 is independently H, lower alkyl, lower alkylamines, COCH 3 , lower alkyl alkenyl, lower alkyl vinyl or lower alkyl aryl. R2 is independently H, OH, halogens, CN, N3, NH2, C(=O)NH2, C(=S)NH2, C(=NH)NH2⋅HCl, C(=NOH)NH2, C(=NH) OMe, lower alkyl, lower alkylamines, lower alkyl alkenyl, lower alkyl vinyl, lower alkyl aryl or substituted heterocyclic compounds;

D is independently selected from CH, CO, N, S, Se, O, NR2, CCONH2, CCH3, C-R2, P or none, where R2 is independently H, O, lower alkyl, lower alkylamines, COCH3, lower alkyl alkenyl, lower alkyl vinyl or lower alkyl aryl, and R2 is independently H, OH, halogens, CN, N3, NH2, lower alkyl, lower alkylamines, lower alkyl alkenyl, lower alkyl vinyl, lower alkyl aryl or substituted heterocyclic compounds;

X is independently O, S, CH 2 or NR; where R is COCH3;

R1 and R4 are independently selected from the group consisting of H, CN, N3, CH2OH, lower alkyl and lower alkyl amines;

R2, R3, R5, R6, R7 and R8 are independently selected from the group consisting of H, OH, CN, N3, halogens, CH2OH, NH2, OCH3, NHCH3, ONHCH3, SCH3, SPh, alkenyl, lower alkyl, lower alkyl amines and substituted heterocyclic compounds; and

R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 and R 8 are not all simultaneously substituted; so that when R2 = R3 = H, then R7 and R8 are hydrogen atoms or none;

when R 1 , R 4 or R 5 are substituted, then R 7 = R 8 = H and R 2 = R 3 = OH;

when R 2 or R 3 are substituted, then R 7 and R 8 are H or OH;

when R 7 or R 8 are substituted, then R 2 and R 3 are H or OH;

when R7 and R8 are hydroxyl, then R2 and R3 are not OH;

when A = N; B = CO; C = N or NH; D = CO or C-NH2; E is CH or C-substituted; F = CH; X = O, S or CH2, then R2 will not be H, OH, CH3, halogen, N3, CN, SH, SPh, CH2OH, CH2OCH3, CH2SH, CH2F, CH2N3, aryl, aryloxy or heterocyclic compound;

when A = N; B = CO; C = N or NH; D = CO or C-NH2; E is CH, C-CH3 or halogen; F = CH; X = N-COCH3, then R2 will not be H or OH;

when A = N; B = CH; C = CH or CH3; D = CH or C-CH3; E is CH, C-CH3 or CCONH2; F = CH; X = O or CH2, then R2 will not be H or OH;

when A = N; B = N, CO or CH; C = CH, C-Cl or C-OCH3; D = CH or C-Ph; E is CH, C-Cl or C-Ph; F = N or CO; X = O, then R 2 will not be H or OH;

when A = N; B = CO or CS; C = N or NH; D = CO or C-NH2; E is CH or N; F = N or CH; X = O, then R 2 will not be H or OH; you

when A = C; B = CH; C = NH; D = CO, CS or C-NH2; E is N or NH; F = CO or CH; X = O, then R 2 will not be H or OH.

Compounds of formula 3 have the following structure:

[image]

where:

X is independently O, S, CH 2 and NR, where R is COCH 3 ;

R' and R" are independently selected from the group consisting of H, CN, C(=O)NH2, NH2, C(=S)NH2, C(=NH)NH2⋅HCI, C(=NOH)NH2, C( =NH)OMe, heterocyclic compounds, halogens, lower alkyl or lower alkyl aryl;

R1 and R4 are independently selected from the group consisting of H, CN, N3, CH2OH, lower alkyl or lower alkyl amines; and

R2, R3, R5, R6, R7 and R8 are independently selected from the group consisting of H, OH, CN, N3, halogens, CH2OH, NH2, OCH3, NHCH3, ONHCH3, SCH3, SPh, alkenyl, lower alkyl, lower alkyl amines or substituted heterocyclic compounds; so

when R2 = R3 = H, then R7 and R8 are hydrogen or none.

In compounds of formula 3, R' is preferably carboxamide or CN and R" is hydrogen or halogens; R1 = R4 = R5 = R7 = R8 = H and R2 = R3 = OH, and X is preferably oxygen.

Compounds of formula 4 have the following structure:

[image]

where:

A is independently selected from N or C;

B, C, E and F are independently selected from the group consisting of CH, CO, N, S, Se, O, NR 2 , CCONH 2 , CCH 3 , C-R 2 or P; R 2 is independently H, lower alkyl, lower alkylamines, COCH 3 , lower alkyl alkenyl, lower alkyl vinyl or lower alkyl aryl.

R2 is independently H, OH, halogens, CN, N3, NH2, C(=O)NH2, C(=S)NH2, C(=NH)NH2⋅HCl, C(=NOH)NH2, C(=NH) OMe, lower alkyl, lower alkylamines, lower alkyl alkenyl, lower alkyl vinyl, lower alkyl aryl or substituted heterocyclic compounds;

X is independently O, S, CH 2 or NR; where R is COCH3;

R1 and R4 are independently selected from the group consisting of H, CN, N3, CH2OH, lower alkyl or lower alkyl amines; you

R2, R3, R5, R6, R7 and R8 are independently selected from the group consisting of H, OH, CN, N3, halogen, NH2, CH2OH, OCH3, NHCH3, ONHCH3, SCH3, SPh, alkenyl, allyl, lower alkyl, lower alkyl amines or substituted heterocyclic compounds; so

when R2 = R3 = H, then R7 and R8 are hydrogen or none;

when A is carbon; B = E = N; C is N-Ph, then F is not CH;

when A = N; C is CH; B = E = C-CH3, then F is not nitrogen; you

when A is carbon, B = N; C = C-CONH2; E = CH; F = S, then X is not CH2.

In compounds of formula 4, R 1 is preferably H, lower alkyl or allyl; R2 is preferably H, OH, halogens, CN, N3, NH2, C(=O)NH2, C(=S)NH2, C(=NH)NH2⋅HCl, C(=NOH)NH2 or C(=NH) OMe; and when R1=R4=R5=R7=R8=H, then preferably R2 = R3 = OH and X is preferably oxygen.

Compounds of formula 5 have the following structure:

[image]

where:

A is independently selected from N or C;

B, C, E and F are independently selected from the group consisting of CH, CO, N, S, Se, O, NR1, CCONH2, CCH3, C-R2 or P; R 1 is independently H, lower alkyl, lower alkylamines, COCH 3 , lower alkyl alkenyl, lower alkyl vinyl or lower alkyl aryl. R2 is independently H, OH, halogens, CN, N3, NH2, C(=O)NH2, C(=S)NH2, C(=NH)NH2⋅HCl, C(=NOH)NH2, C(=NH) OMe, lower alkyl, lower alkylamines, lower alkyl alkenyl, lower alkyl vinyl, lower alkyl aryl or substituted heterocyclic compounds;

D is independently selected from the group consisting of CH, CO, N, S, Se, O, NR1, CCONH2, CCH3, C-R2, P or none; R 1 is independently H, O, lower alkyl, lower alkylamines, COCH 3 , lower alkyl alkenyl, lower alkyl vinyl or lower alkyl aryl. R2 is independently H, OH, halogens, CN, N3, NH2, lower alkyl, lower alkylamines, lower alkyl alkenyl, lower alkyl vinyl, lower alkyl aryls or substituted heterocyclic compounds;

X is independently O, S, CH 2 or NR where R is COCH 3 ;

R1 and R4 are independently selected from the group consisting of H, CN, N3, CH2OH, lower alkyl and lower alkyl amines; you

R2, R3, R5, R6, R7 and R8 are independently selected from the group consisting of H, OH, CN, N3, halogens, CH2OH, NH2, OCH3, NHCH3, ONHCH3, SCH3, SPh, alkenyl, lower alkyl, lower alkyl amines and substituted heterocyclic compounds; so

when R2 = R3 = H, then R7 and R8 are hydrogen or none.

when A = N; B = CO; C = N or NH; D = CO or C-NH2; E is CH or C-substituted; F = CH; X = O, S or CH2, then R2 will not be H, OH, CH3, halogens, N3, CN, SH, SPh, CH2OH, CH2OCH3, CH2SH, CH2F, CH2N3, aryl, aryloxy or heterocyclic.

when A = N; B = CO; C = N or NH; D = CO or C-NH2; E is CH, C-CH3 or halogen; F = CH; X = N-COCH3, then R2 will not be H or OH;

when A = N; B = CH; C = CH or CH3; D = CH or C-CH3; E is CH, C-CH3 or CCONH2; F = CH; X = O or CH2, then R2 will not be H or OH;

when A = N; B = N, CO or CH; C = CH, C-Cl or C-OCH3; D = CH or C-Ph; E is CH, C-Cl or C-Ph; F = N or CO; X = O, then R 2 will not be H or OH;

when A = N; B = CO or CS; C = N or NH; D = CO or C-NH2; E is CH or N; F = N or CH; X = O, then R 2 will not be H or OH; you

when A = C; B = CH; C = NH; D = CO, CS or C-NH2; E is N or NH; F = CO or CH; X = O, then R 2 will not be H or OH.

In a further aspect of the present invention, an abnormality expressed by an increased response in one group of cytokines is treated with the administration of a nucleoside or other compound that increases the response in another group of cytokines. Thus, for example, the usual sudden onset of allergy results in an abnormally increased Th2 response. This abnormality is treated with Ribavirin between 600 mg/day and 1000 mg/day (for a typical adult), inducing a Th1 response. The treatment is effective because Th1 and Th2 have an increase-decrease type of relationship in this case, so the Th2 response is suppressed.

In a further aspect of this invention, the patient is treated prophylactically with the use of a nucleoside or other compound that selectively reduces Th1 activity without significantly reducing Th2 activity. Prophylaxis can, for example, prepare the patient for a tissue or organ transplant, or for anticipated contact with allergens.

In a further aspect of the present invention, the nucleoside or other compound is administered to the patient at a dose that reduces the total amount of GTP in the patient to a degree that selectively reduces one of the Th1 or Th2 responses, without significantly reducing the other response. Controlled release dosage forms in particular are believed to achieve this result, particularly formulations that maintain the serum dose of the compound within a desirable range. In the case of Ribavirin, for example, the serum level should be between about 2 μM and about 5 μM. Regarding delivery rates, a controlled-release formulation may ideally have an in vitro dissolution rate measured by the USP "paddle method" at 100 rmp for 900 ml of aqueous buffer (pH between 1.6 and 7.2) between about 15% and 40% ( wt.) of the compound after 1 hour, between about 30% and about 50% (wt.) of the compound after 2 hours, about 50% and 70% (wt.) of the compound after 4 hours, between about 60% and about 80% (wt .) date after 6 hours.

Use

It is believed that the combinations set forth in the claims will be used to treat a whole range of conditions, in fact any condition that responds favorably to the application of one or more such combinations. Among other things, it is specifically assumed that such combinations will be able to be used to treat infection, attacks, tumors, increased sensitivity or autoimmune disease.

Infections believed to be treatable with the compounds of this invention include respiratory syncytial virus (RSV), hepatitis B virus (HBV), hepatitis C virus (HCV), herpes simplex types 1 and 2, herpes genitalis, herpes keratitis, herpes encephalitis, herpes zoster, human immunodeficiency virus (HIV), influenza A virus, Hantan virus (hemorrhagic fever), human papilloma virus (HPV), measles and leprosy. In particular, the combinations set forth in the claims are believed to be useful in the treatment of chronic bacterial and viral infections, including HIV, tuberculosis, leprosy, and others.

Infestations are considered treatable with the compounds of this invention if they involve intracellular protozoan attack, intestinal worms, or other parasitic infections. Again, it is particularly believed that the combinations set forth herein in the claims will be useful in the treatment of chronic infestations.

It is believed that those tumors that are caused by the virus can be treated, and that the action can include inhibiting the transformation of the cells infected by the virus into a neoplastic state, stopping the spread of the virus from the transformed cells to other normal cells and/or stopping the growth of the cells that have been changed by the virus.

Treatable hypersensitivity includes all types of allergy, including IgE and IgG allergies, hyper IgE syndrome, and skin conditions such as atopic dermatitis. It is also believed that the combinations set forth in the claims can be used to treat transplant rejection (recipient organ-host relationship) and implant reaction.

Autoimmune diseases can be classified as organ-nonspecific and organ-specific. Organ-nonspecific autoimmune diseases include rheumatoid arthritis, gout and gouty arthritis, systemic lupus erythematosus (SLE), Sjögren's syndrome, scleroderma, polymyositis and dermatomyositis, ankyloid spondylitis, and rheumatic fever. Organ-specific autoimmune diseases are known for almost every organ, including insulin-dependent diabetes, thyroid diseases (Graves' disease and Hashimoto's thyroiditis), Addison's disease, and some kidney and lung diseases including allergy and asthma, multiple sclerosis, myasthenia gravis, uveitis, psoriasis, forms of hepatitis and cirrhosis, celiac disease, inflammatory abdominal diseases, and some types of male and female sterility. Autoimmune diseases can also be triggered by viral infections including the HIV virus, can occur as a result of transplant rejection, can accompany some tumors, or occur after exposure to some chemicals.

It is also hypothesized that an abnormality reflected by an increased response of one group of cytokines can be treated by the administration of a nucleoside that increases the response in another group of cytokines. So, for example, since common IgE allergies are associated with a predominant Th2 response, they can be treated with Ribavirin, which increases the Th1 response at a low dose of about 500 mg/day to about 1,000 mg/day.

In a further aspect of this invention, the patient is treated prophylactically with a compound that selectively reduces Th1 activity without significantly reducing Th2 activity. Prophylactic treatment can reduce the expected adverse effects of expected events, such as tissue or organ transplantation, or reduce symptoms for an expected pulmonary insult, such as an increase in pollen levels in the air during spring.

Synthesis

The synthesis of compounds of formula 1, 1-A through 1-F, is described in PCT application PCT/LIS97/18387, which is incorporated herein in its entirety. Synthesis of compounds according to formulas 2 to 5 is described in PCT application PCT/US97/00600.

Application

It is anticipated that the compounds of this invention will be administered in the form of any suitable pharmaceutical formulation, and under the conditions of any suitable protocol. When primary or “first” treatment is used as described above, preferred monotherapeutic doses and protocols for such drugs are described in the PDR, or at least provided by the manufacturer or distributor. Preferred dosages and protocols for a “second” drug, such as the bimodal nucleosides described herein, can be determined by experimentation with the particular patient. This experimentation need not be exhaustive, and it is anticipated that the "second" drug containing the nucleosides described herein will be administered at a dose between about 100 mg/day and about 5000 mg/day.

Of course, those familiar with this field know that the therapeutically effective amount varies depending on the infection or condition to be treated, the severity of the disease, the treatment regimen being applied, the pharmacokinetics of the agent being used, as well as the patient (animal or human) being treated. Thus, an effective dose may vary from 1 mg/kg body weight, or less, to 25 mg/kg body weight or more. In general, a therapeutically effective amount of the "second" drug is believed to be in the range of slightly less than 1 mg/kg to about 25 mg/kg, depending on the compound being used, the condition or infection being treated, and the route of administration. The dosage range can generally provide an effective blood concentration of the active compound that is in the range of about 0.04 to about 100 micrograms/cm 2 of patient blood. It is believed, however, that appropriate patient-specific regimens can be developed using a small amount, which is then increased until undesirable side effects are observed or until the desired effect is achieved.

Administration of the compounds according to this invention can be done orally, parenterally (including subcutaneous injections, intravenous, intramuscular, and intrasternal injections or infusion techniques), in inhalation spray, rectally, topically or otherwise, and in unit dosage formulations containing conventional pharmaceutical acceptable carriers, adjuvants and transfer agents.

It is believed that the compounds of the present invention may be formulated in admixture with a pharmaceutically acceptable carrier. For example, the compounds of this invention can be administered orally in the form of pharmaceutically acceptable salts. Since the compounds of this invention are mostly water soluble, they can be administered intravenously in saline (eg, buffered to a pH of 7.2 to 7.5). Common buffers such as phosphates, bicarbonates or citrates can be used for this purpose. Of course, one skilled in the art can modify the formulations within the specifications to obtain numerous formulations for a particular route of administration, without rendering the composites of this invention unstable or altering their therapeutic activity. In particular, modifications of these compounds that will make them more soluble in water or other media, for example, can be easily achieved by small changes (salt formation, esterification, etc.) as is well known by those skilled in the art. Those skilled in the art also know that the route of administration or dosage regimen of a particular compound may be altered to manipulate the pharmacokinetics of these compounds to achieve the greatest benefit in the patient.

For some pharmaceutical dosage forms, the prodrug form of the administered compound, especially including acylated (acetylated or other) derivatives, pyridine esters and various salt forms of the compounds listed herein are preferred. Those skilled in the art know how the compounds herein can be easily modified into pro-drug forms to facilitate delivery of the active compounds to the target site within the host or patient organism. Those skilled in the art may also take advantage of the preferred pharmacokinetic parameters of the prodrug form, when possible, in delivering a compound herein to a target site within the host or patient to maximize the desired effect of the compound.

Furthermore, the compounds included in the combination according to the present invention can be administered separately or together, and if administered separately, it can be done in any order. The amounts of active additives (active additive) and pharmaceutical active substance(s) and the relative timing of administration will be selected to achieve the desired combined therapeutic effect.

The method of administration of the compounds according to this invention may range from continuous (intravenous drip) to several oral administrations per day (for example Q.I.D.) and may include oral, topical, parenteral, intramuscular, intravenous, subcutaneous, transdermal (which may include a penetration enhancer ), buccal administration or suppository administration, among other ways of administration.

To prepare therapies according to the present invention, a therapeutically effective amount of the compound is preferably intimately mixed with a pharmaceutically acceptable carrier according to conventional pharmaceutical binding techniques to form a dosage. The carrier can take a wide variety of forms depending on the form of preparation desired for administration, eg oral or parenteral. Any of the usual pharmaceutical media can be used to prepare the pharmaceutical compositions in oral form. Accordingly, for liquid oral preparations such as suspensions, elixirs and solutions, suitable carriers and additives including water, glycols, oils, alcohols, flavor enhancers, preservatives, coloring agents and the like can be used. For solid oral preparations such as powders, tablets and capsules, and for solid preparations such as suppositories, suitable carriers and additives can be used including starch, sugar carrier such as dextrose, mannitol, lactose and related carriers, diluents, agents for granulation, lubricants, binders, disintegrants and the like. If desired, tablets and capsules may be enteric-coated or delayed-release, made by standard techniques.

For parenteral formulations, the carrier will usually comprise sterile water or aqueous sodium chloride, although other dispersing agents may be included. Of course, when sterile water is used, which should remain sterile, composites and supports must also be sterilized. Injectable suspensions can also be prepared, when suitable liquid carriers, suspending agents and the like can be employed.

It is also generally contemplated that the most preferred uses of the present invention are those for which the active compounds are relatively less cytotoxic to non-target host cells and relatively more active to target cells. Accordingly, it is advantageous that L-nucleosides can be of increased stability compared to D-nucleosides, which can provide better pharmacokinetics. This effect can be achieved because the enzymes will not recognize L-nucleosides, and accordingly they can have a longer half-life.

Therefore, therapies have been described in which nucleosides and other compounds are used to selectively modulate Th1 and Th2 responses relative to each other in the treatment of disease. Although specific embodiments are described herein, the scope of this invention is not limited except as explained in the appended claims.

Claims (51)

1. A method for reducing the administered dose of the first drug in the treatment of a disease known to cause an abnormality of at least one cytokine in a patient, indicated by this, which includes: - identifying the monotherapy dosage of the first drug that is effective in the treatment of the disease; - identifying another drug as one that worsens the irregularity when used as monotherapy within the dosage range; you - application of combined therapy that contains the first drug less than in monotherapy dosing and the second drug that is outside the dosage range. 2. The method according to claim 1, characterized in that the disease includes a chronic disease. 3. The method according to claim 1, characterized in that the disease includes a chronic viral disease. 4. The method according to claim 1, characterized in that the disease includes insulin-dependent diabetes. 5. The method according to claim 1, characterized in that the disease includes an allergy. 6. The method according to claim 1, characterized in that the disease includes atopic dermatitis. 7. The method according to claim 1, characterized in that the disease involves intracellular protozoan infection. 8. The method according to claim 1, characterized in that the disease includes hyper IgE syndrome. 9. The method according to claim 1, characterized in that the disease includes HIV. 10. The method of claim 1, wherein the disease comprises an organ rejection disease. 11. The method according to claim 1, characterized in that the disease includes systemic Lupus Erythematosus. 12. The method according to claim 1, characterized in that the disease includes a tumor. 13. The method according to any one of claims 1-12, characterized in that the abnormality comprises an abnormal increase in Th1 activity. 14. The method according to any one of claims 1-12, characterized in that the abnormality comprises an abnormal reduction of Th1 activity. 15. The method according to any one of claims 1-12, characterized in that the abnormality comprises an abnormal increase in Th2 activity. 16. The method of any one of claims 1-12, wherein the abnormality comprises an abnormal reduction in Th2 activity. 17. The method according to any one of claims 1-12, characterized in that the second drug comprises a pharmaceutically acceptable form of nucleoside. 18. The method of any one of claims 1-12, wherein the second drug comprises a pharmaceutically acceptable form of D-nucleoside. 19. The method according to any one of claims 1-12, characterized in that the second drug comprises a pharmaceutically acceptable form of L-nucleoside. 20. The method according to any one of claims 1-12, characterized in that the second drug includes a pharmaceutically acceptable form of Ribavirin. 21. The method according to any one of claims 1-12, characterized in that the second drug comprises a pharmaceutically acceptable form of interferon. 22. The method according to any one of claims 1-12, characterized in that the second drug includes a nucleoside according to at least one of formulas 1, 1A, 1B, 1C, 1D, 1E or 1F. 23. The method according to any one of claims 1-12, characterized in that the second drug includes a nucleoside according to at least one of the formulas 2, 3, 4 or 5. 24. Method for prophylactic treatment of patients, indicated by this, which includes: - obtaining pharmaceuticals that suppress Th1 activity in patients when applied above the given dosage level; you - administration of pharmaceuticals to the patient below the given dosage level. 25. The method according to claim 24, characterized in that the prophylaxis includes preparing the patient for organ transplantation. 26. The method according to claim 24, characterized in that the prophylaxis includes preparing the patient for tissue transplantation. 27. The method according to claim 24, characterized in that the prophylaxis includes preparing the patient for expected contact with allergens. 28. The method of any one of claims 24-27, further comprising a pharmaceutical at a dose that induces Th2 activity. 29. The method according to any one of claims 24-27, characterized in that the pharmaceutical includes a nucleoside according to at least one of formulas 1, 1A, 1B, 1C, 1D, 1E or 1F. 30. The method according to any one of claims 24-27, characterized in that the pharmaceutical includes a nucleoside according to at least one of the formulas 2, 3, 4 or 5. 31. A method for treating a disease characterized by an increased or decreased Th1 or Th2 response, characterized by: - identifying a compound and a dosage range for a compound that is effective in reducing the total amount of GTP and thereby selectively reduces one of the Th1 or Th2 responses without significantly reducing the other response; you - administration of nucleosides to the patient within the dosage range. 32. The method of claim 31, wherein the disease comprises a chronic disease. 33. The method of claim 31, wherein the disease comprises a chronic viral disease. 34. The method according to claim 31, characterized in that the disease includes insulin-dependent diabetes. 35. The method according to claim 31, characterized in that the disease includes an allergy. 36. The method according to claim 31, characterized in that the disease includes atopic dermatitis. 37. The method according to claim 31, characterized in that the disease involves intracellular protozoan infection. 38. The method according to claim 31, characterized in that the disease includes hyper IgE syndrome. 39. The method of claim 31, wherein the disease includes HIV. 40. The method according to claim 31, characterized in that the disease involves organ rejection. 41. The method according to claim 31, characterized in that the disease includes systemic Lupus Erythematosus. 42. The method according to claim 31, characterized in that the disease includes a tumor. 43. The method according to any one of claims 31-42, characterized in that the compound contains a nucleoside according to formulas 1, 1A, 1B, 1C, 1D, 1E or 1F. 44. The method according to any one of claims 31-42, characterized in that the compound contains a nucleoside according to formulas 2, 3, 4 or 5. 45. A controlled-release formulation for oral administration, comprising a compound effective in selectively modulating Th1 and Th2 responses relative to each other within a dosage range, wherein the controlled-release aspect helps maintain serum levels below those that suppress Th1 and Th2 responses . 46. Compound with controlled release according to claim 45, characterized in that it contains a nucleoside according to at least one of formulas 1, 1A, 1B, 1C, 1D, 1E or 1F. 47. Compound with controlled release according to claim 45, characterized in that it contains a nucleoside according to at least one of the formulas 2, 3, 4 or 5. 48. A compound with controlled release according to claim 45, characterized in that it contains Ribavirin. 49. A compound with controlled release according to claim 45, characterized in that it contains an interferon. 50. The controlled release composition of any one of claims 45-49, wherein the patient maintains a serum level between about 2 μM to about 5 μM of the compound. 51. The controlled release composition according to any one of claims 45-49, characterized in that the composition has an in vitro dissolution rate measured by the USP "paddle method" of 100 rpm for 900 ml of aqueous buffer (pH between 1.6 and 7.2) between about 15% and 40% (wt.) of the compound after 1 hour, between about 30% and about 50% (wt.) of the compound after 2 hours, about 50% and 70% (wt.) of the compound after 4 hours, between about 60 % and about 80% (wt.) of the compound after 6 hours.