MXPA99001961A - Use of pkc inhibitors for the manufacture of a medicament for the treatment of htlv-1 infections - Google Patents

Abstract

A method for treating human T cell lymphotrophic virus type 1 infection using an isozyme selective PKC inhibitor, particularly using the isozyme selective PKC inhibitor, (S)-3,4-[N,-N'-1, 1'-((2''-ethoxy)-3'''(O)-4'''-(N,N-dimethylamino)-butane)-bis-(3,3'-indolyl)]-1(H)-pyrrole-2,5-dione hydrochloride salt.

Description

USE OF PKC INHIBITORS FOR MANUFACTURING A MEDICINE FOR THE TREATMENT OF HTLV-1 INFECTIONS BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention is broadly directed to a method of treating infection with type 1 human T cell lymphotropic virus (HTLV-1). In particular, the invention is directed to a method for inhibiting the induced effects of the HTLV-1 viral protein activity such as Tax 1 activity and to inhibit the replication of HTLV-1. The present invention is particularly directed to the use of a particular class of Protein Kinase C (PKC) inhibitors selective for isozyme to treat HTLV-1 infection and diseases associated therewith such as T-cell leukemia and central nervous system disorder induced by HTLV-1. 2. Description of the Related Art People infected with the human T-cell lymphotropic virus type 1 (HTLV-1) are prone to develop adult T cell leukemia. In addition, individuals infected with HTLV-1 may manifest chronic neurodegenerative diseases such as tropical spastic paraparesis and myelopathy. No particular treatment for HTLV-1 infection is available in the art. Treatment for diseases associated with HTLV-1 infection is symptomatic and does not provide satisfactory results. Therefore, there is a need in the subject of developing therapeutic agents for the treatment of HTLV-1 infection and the diseases associated therewith. Leukemia is a disease characterized by neoplastic proliferation of one of the blood-forming cells. The different types of leukemia are classified according to the type of cell involved and as acute or chronic, depending on the duration of the disease. If left untreated, all forms of leukemia are fatal. Death is usually due to complications resulting from infiltration into the bone marrow by leukemic cells and the replacement of normal hematopoietic cells. Adult T cell leukemia can be associated with HTLV-1 infection. During the last quarter of a century, a worldwide effort has been made to improve the treatment of leukemia. Using the best current treatment regimens, more than 90 percent of children with acute lymphoblastic leukemia (ALL) now achieve complete remission. However, adults with ALL, especially T cell leukemia, generally respond less favorably to treatment than children, and most trials have only resulted in complete remission regimens of 50 percent or less and only with an average duration one year or less. Therefore, there is a continuing need in the art to develop new therapeutic agents for the treatment of adult T cell leukemia associated with HTLV-1 infection. HTLV-1 infection can also cause chronic neurodegenerative disorders. Chronic neurodegenerative diseases such as tropical spastic paraparesis and myelopathy can cause loss of function, suppression of reflex activity and other complications. In general, the treatment of neurodegenerative disorders is conservative and symptomatic without complete recovery. Therefore, there is also a need in the art to develop new therapeutic agents for the treatment of chronic neurodegenerative disorders associated with HTLV-1 infection. SUMMARY OF THE INVENTION An objective of the invention, therefore, provides a method for inhibiting the replication of human T cell lymphotrophic virus type 1 in an infected cell. It is another object of the invention to provide a method for inhibiting an effect of the viral protein activity of Tax 1 in a cell. It is still another object of the invention to provide a method for treating a mammal infected with a virus lymphotrophic human T-cell type 1 It is still another object of the invention to provide a method for treating adult T cell leukemia associated with infection of human T-cell lymphotropic virus type 1. It is still another object of the invention to provide a method for treating a chronic neurodegenerative disorder associated with the infection of type 1 human T-cell lymphotropic virus. These and other objects of the invention are provided by one or more of the modalities described below. One embodiment of the invention provides a method for inhibiting the replication of human T cell lymphotrophic virus type 1 in a cell comprising contacting the cell with an inhibitory amount of viral replication of a protein kinase C inhibitor. Another embodiment of the invention provides a method for inhibiting the effect of the viral protein activity Tax 1 in a cell comprising in contact with the cell with a viral protein activity that inhibits the amount of a protein kinase C inhibitor. Yet another embodiment of the invention provides a method for treating an animal infected with human T cell lymphotropic virus type 1 comprising administering to the mammal a therapeutically effective amount of the protein kinase C inhibitor. Yet another embodiment of the invention provides a method of treating adult T cell leukemia associated with infection with human T-cell lymphotropic virus type 1 comprising administering to an adult in need said treatment a therapeutically effective amount of a selective inhibitor of the isozi protein beta kinase beta C. Yet another embodiment of the invention provides a method for treating a chronic neurodegenerative disorder associated with infection with type 1 human T-cell lymphotropic virus which comprises administering to the mammal a therapeutically effective amount of a selective isozyme inhibitor. beta protein kinase C. Therefore, the present invention provides the subject with compounds effective to treat HTLV-1 infection and diseases associated therewith. DETAILED DESCRIPTION OF THE INVENTION It is a discovery of the present invention that the therapeutic use of a particular class of protein kinase C inhibitors, ie inhibitors of the protein isozyme β of protein kinase C and especially selective inhibitors of the β-isozyme of PKC, inhibits the effects induced by HTLV- 1 of viral protein activity and particularly viral replication. Consequently, said compounds can be used therapeutically to treat patients infected with HTLV-1 infection. HTLV-1 replication comprises the multiplication of the HTLV-1 genome during infection and productive reactivation of latent infection. The reactivation of the latent genome of integrated HTLV includes replication of HTLV, e.g. , form multiple transcripts of the HTLV-1 genome, the appropriate process of transcripts of HTLV-1, expression of HTLV-1, e.g. , translation of virus-specific proteins such as Tax 1, similarity of the virus, as well as release of infectious HTLV-1 particles and HTLV-1 proteins. HTLV-1 viral proteins such as Tax 1 exert their effects via interaction with the viral system as well as the host system. The effects induced by HTLV-1 viral protein activity include but are not limited to regulating the transcription of the HTLV-1 virus as well as a variety of other cellular genes (Kwanyee et al., Nature 23: 776-778, 1988; et al., Mol. Cell, Biol. 9: 1733-1745, 1989, and Yoshimura et al., EMBO J 9: 2537-2542, 1990). The viral protein Tax 1 is secreted extracellularly by cells infected with HTLV-1 (Lindholm et al., New Biol. 2: 1034-1043, 1990). Extracellular Tax 1 secreted can be absorbed by surrounding cells and activate transcriptional factors such as NF-? B in cells (Smith et al., J. Clin.Research 87: 761-766, 1991; Ari et al., J. Virology 65: 6892-6899, 1991). It has been shown that cells infected with HTLV-1 produce certain constitutively transcriptional factors such as NF-? B (Lindholm et al., J Virology 70: 2525-2532, 1996). Individuals infected with HTLV-1 are prone to develop adult T cell leukemia (Poiesz et al., PNAS 77: 7415-7419, 1980; Hinuma et al., PNAS 78: 6476-6480, 1981). In addition, individuals infected with HTLV-1 may manifest chronic neurodegenerative disorders such as tropical spastic paraparesis and myelopathy (Gessain et al., Lancet I I: 407-410, 1985, Osame et al., Lancet I: 1031-1032, 1986). Although not wishing to be limited by any technical explanation, the applicants think that PKC affects HTLV-1 viral protein activity and, therefore, viral replication. The viral protein Tax 1 interacts directly with and activates PKC (Lindholm et al., J Virology 70: 2525-2532, 1996). It has been shown that some PKC inhibitors block Tax 1 -induced activation of cellular proteins such as N F-? B in cells infected with HTLV-1 (Lindholm et al., J Virology 70: 2525-2532, 1996). It has also been shown that inhibition of PKC blocks the proper processing of HTLV-1 mRNA that is required for efficient viral application (Adchi et al., Biochem Biophys, Res. Commun. 169: 469-475, 1990). The additional evidence supporting the role of PKC in the efficient replication of HTLV-1 is the ability of PKC inhibitors to decrease the expression of bovine leukemia virus that is structurally and biologically similar to HTLV-1 (Jensen et al., J Virology 66: 4427-4433, 1992). Therefore, the compounds of PKC inhibitors as described in the present invention can be used therapeutically to treat HTLV-1 infection and the diseases associated therewith by suppressing viral protein activity and especially viral replication. The method of this invention preferably utilizes those protein kinase C inhibitors that effectively inhibit the β-isozyme. A suitable group of compounds is generally described in the prior art as bis-indolylmaleimides or macrocyclic bis-indoylmaleimides. The bis-indolylmaleimides well recognized in the prior art include those compounds described in the Patents of E.U.A. Nos. 5,621,098, 5,552,396, 5,545,636, 5,481, 003, 5,491, 242 and 5,057,614, incorporated herein by reference. The macrocyclic bis-indolylmaleimides are in particular represented by the compounds of the formula I. These compounds and methods for their preparation have been described in the U.A. No. 5,552,396, which is incorporated herein by reference. These compounds are administered in a therapeutically effective amount for a human in order to inhibit the induced effects of HTLV-1 viral protein activity such as Tax 1 activity and to inhibit the replication of HTLV-1, or to treat HTLV-1 infection. These compounds can also be administered to patients who are at risk for the disease conditions mentioned above as prophylactics.

A preferred class of compounds for use in the method of the invention has the formula (I) wherein: W is -O-, -S-, -SO-, -S02-, -CO-, C-C6 alkylene, substituted alkylene, C2-C6 alkenylene, -aryl, -aryl (CH2) mO- , -heterocycle-, heterocycle- (CH2) m-, bicyclic fused-, -cyclic fused- (CH2) mO, -N R3-, -ÑOR3-, -CON H-, or -N HCO-; X and Y are independently C? -C alkylene, substituted alkylene, or together X, Y, and W combine to form - (CH2) n-AA-; R1 are hydrogen or up to four optional substituents independently selected from halo, C? -C4 alkyl, hydroxy, C? -C4 alkoxy, haloalkyl, nitro, -NR4R5, or -N HCO (C? -C4 alkyl); R2 is hydrogen, CH3CO-, -N H2 or hydroxy; R3 is hydrogen, - (CH2) maryl, C? -C alkyl, -COO (dC4 alkyl), -CONR4R5, - (C = NH) N H2, -SO (C? -C4 alkyl) ), -S02 (NR4R5) or -S02 (C? -C4 alkyl); R4 and Rs are independently hydrogen, C1-C4 alkyl, phenyl. benzyl or combines with the nitrogen which are joined to form a saturated or unsaturated 5 or 6 membered ring; AA is an amino acid residue; m is independently 0, 1, 2, or 3; and n is independently 2, 3, 4, or 5, or a pharmaceutically acceptable salt, prodrug or ester thereof. A more preferred class of compounds for use in this invention is represented by the formula I wherein the -X-W-Y-portions contain from 4 to 8 atoms, which may be substituted or unsubstituted. More preferably, the -X-W-Y- portions contain 6 atoms. Other preferred compounds for use in the method of this invention are those of the formula I wherein R1 and R2 are hydrogen; and W is a substituted alkylene, -O-, S-, -CONH-, -N HCO- or -NR3-. Particularly preferred compounds for use in the invention are compounds of formula 1 a: wherein Z is - (CH2) P- or - (CH2) p-O- (CH2) p-; R4 is hydroxy, -SH, d-C4 alkyl, (CH2) maryl, -N H (aryl), -N (CH3) (CF3) -N H (CF3), or N R5R6; R 5 is hydrogen or C 1 -C 4 alkyl; R6 is hydrogen, C1-C4 alkyl or benzyl; p is 0, 1 or 2; and m is independently 2 or 3, or a pharmaceutically acceptable salt, prodrug or ester thereof. The most preferred compounds of formula 1 a are those wherein Z is CH 2; and R 4 is -NH 2, -NH (CF 3), or -N (CH 3) 2 or a pharmaceutically acceptable salt, prodrug or ester thereof. Other preferred compounds for use in the method of the present invention are compounds wherein W in formula I is -O-, Y is a substituted alkylene and X is an alkylene. These preferred compounds are represented by the formula Ib. wherein Z is - (CH2) P-; R 4 is -N R 5 R 6, -N H (CF) or -N (CH 3) (CF), R 5 and R 6 are independently H or C 1 -C 4 alkyl; p is 0, 1 or 2; and m is independently 2 or 3, or a pharmaceutically acceptable salt, prodrug or ester thereof. The most preferred compounds of formula I b are those wherein p is 1; and Rs and R6 are methyl. Because they contain a basic portion, the compounds of formulas I, la and Ib can exist as pharmaceutically acceptable acid addition salts. The acids commonly used to form the salts include inorganic acids such as hydrochloric, hydrobromic, hydroiodic, sulfuric and phosphoric acids, as well as organic acids such as para-toluenesulfonic, oxalic, para-bromophenylsulphonic, carbonic, succinic, citric, benzoic, acetic acids. and related inorganic and organic acids. Said pharmaceutically acceptable salts, therefore, include sulfate, pyrosulfate, bisulfate, sulfite, bisulfite, phosphate, mono-hydrogen phosphate, dihydrogen phosphate, metaphosphate, pyrophosphate, chloride, bromide, iodide, acetate, propionate, decanoate, caprylate, acrylate, format, isobutyrate, heptanoate, propiolate, oxalate, malonate; succinate, suberate, sebacate, fumarate, maleate, 2-butyne-1,4-dioate, 3-hexin-2,5-dioate, benzoate, chlorobenzoate, hydroxybenzoate, methoxybenzoate, phthalate, xylene sulfonate, phenylacetate, phenylpropionate, phenylbutyrate, citrate, lactate, hippurate, β-hydroxybutyrate, glycolate, maleate, tartrate, methanesulfonate, propanesulfonate, naphthalene-1-sulfonate, naphthalene-2-sulfonate, mandelate and the like. In particular, the hydrochloric and mesylate salts are used. In addition to pharmaceutically acceptable salts, other salts may also exist. They can serve as intermediates in the purification of the compounds, in the preparation of other salts or in the identification and characterization of the compounds and intermediates. The pharmaceutically acceptable salts of the compounds of the formulas I, la and Ib may also exist as various solvates, such as with water, methanol, ethanol, dimethylformamide, ethyl acetate and the like. Mixtures of said solvates can also be prepared. The source of said solvate can come from the crystallization solvent, it can be inherent in the preparation or crystallization solvent or adventitious with said solvent. It is recognized that various stereoisomeric forms of the compounds of formulas I, la and I b may exist; for example, W may contain a chiral carbon atom in the substituted alkylene moiety. The compounds are normally prepared as racemates and conveniently can be used as such. Alternatively both individual enantiomers can be isolated or synthesized by conventional techniques if desired.

Said racemates and individual enantiomers and mixtures thereof are part of the compounds used in the methods of the present invention. The compounds used in this invention also encompass pharmaceutically acceptable drugs of the compounds of formulas I, la and Ib. A prodrug is a prodrug that can be chemically modified and may be biologically inactive at its site of action, but which can be degraded or modified by one or more enzymatic processes or other processes in vivo to the mother's bioactive form. This prodrug may probably have a different drug-kinetic profile than the mother form, allowing easier absorption through the mucosal epithelium, better salt formation or solubility and / or improved systemic stability (an increase in plasma half-life, by example). Typically, said chemical modifications include the following: 1) ester or amide derivatives that can be separated by esterases or lipases; 2) peptides that can be recognized by specific and non-specific proteases; or 3) derivatives that accumulate at a site of action through membrane selection of a prodrug form or a modified prodrug form; or any combination of 1 to 3, supra. Conventional procedures for the selection and preparation of suitable prodrug derivatives are described, for example in, H. Bundgaard, Design of Prodrugs. (1985).

The synthesis of various bis-indole-N-maleimide derivatives are described in Davis et al., Patent of E.U.A. No. 5,057,614 and the synthesis of preferred compounds suitable for use in this invention are described in the Patents of E.U.A. previously identified 5,552,396 and Faul et al. EP 0 657 41 1 A1, all of which are incorporated herein by reference. A preferred protein kinase inhibitor for use in the method of this invention is the compound described in Example 5g hydrochloride salt of ((S) -3,4- [N, N -'- 1, 1 '- ( (2"-ethoxy) -3 '" (0) -4' "- (N, N-dimethylamino) -butane) -bis (3,3'-indolyl)] - 1 - (H) -pyrrol-2, 5-dione of US Patent 5,552,396 mentioned above.This compound is a potent inhibitor of protein kinase C. It is selective for protein kinase C on other kinases and is highly isozyme selective, ie it is selective for beta-1 isozymes. and beta 2. Other salts of this compound could also be favored, especially the mesylate salts.A preferred mesylate salt can be prepared by reacting a compound of the formula II. with the methanesulfonic acid in a non-reactive organic solvent, preferably an organic / water mixture, and more preferably water-acetone. Other solvents such as methanol, acetone, ethyl acetate and mixtures thereof are also operable. The ratio of the solvent to water is not critical and is generally determined by the solubility of the reagents. The preferred solvent to water ratios are generally from 0.1: 1 to 100: 1 solvent to water by volume. Preferably, the ratio is 1: 1 to 20: 1 and more preferably 5: 1 to 10: 1. The optimum ratio depends on the selected solvent and preferably is a solvent ratio of acetone to water of 9: 1. The reaction usually involves approximately equimolar amounts of two reactants, although other ratios are operative, especially those in which methanesulfonic acid is in excess. The methanesulfonic acid addition rate is not critical to the reaction and can be added quickly (<5 minutes) or slowly for 6 or more hours. The reaction is carried out at temperatures ranging from 0 ° C to reflux. The reaction mixture is stirred until Complete the formation of the salt, as determined by the powder diffraction of x-rays and can take from 5 minutes to 12 hours. The salts of the present invention are preferably and easily prepared as a crystalline form. The trihydrate form of the salt can easily be converted to the monohydrate when dried or exposed to 20-60% relative humidity. The salt is substantially crystalline demonstrating a defined melting point, birefringence and a pattern of x-ray diffraction. Generally, the crystals have less than 10% amorphous solids and preferably less than 5% and still more preferably less than 1% amorphous solids. The mesylate salt is isolated by filtration or other separation techniques appreciated in the art, directly from the reaction mixture in yields ranging from 50% to 100%. Recrystallization and other purification techniques known in the art can be used to further purify the salt if desired. One skilled in the art will recognize that a therapeutically effective amount of the protein kinase C inhibitor of the present invention is the amount sufficient to inhibit the replication of HTLV-1 or inhibit the activity of viral protein such as Tax 1. It is within the ability of a person skilled in the art to measure the re-delivery of HTLV-1 and viral protein activity such as Tax 1 activity. The amount administered varies among other things, depending on the concentration of the compound in the therapeutic formulation and the patient's body weight. Generally, an amount of protein kinase C inhibitor that will be administered as an agent will be determined. therapeutic couple to treat HTLV-1 infection on a case-by-case basis by the doctor. As a guideline, the degree of infection, the resistance of the immune system, the number of leukemic cells, the viral load, the body weight and the age of the patient will be considered when an appropriate dose is established. Generally, a suitable dose is one which results in a concentration of the protein kinase C inhibitor in the treatment site in the range of 0.5 mM to 200 μM and more usually 0.5 nM to 200 nM. It is expected that serum concentrations of 0.5 nM to 20 nM will be sufficient in most circumstances. To obtain these treatment conditions, a patient in need of treatment will probably be administered between approximately 0.001 mg per day per kg. of body weight and 50.0 mg per day per kg. Usually, no more than about 10.0 mg per day per kg will be necessary. of body weight of protein kinase C inhibitor. As noted above, the above amounts may vary on a case-by-case basis. The effectiveness of the compounds of the invention can be evaluated in various experimental characteristics readily available in the art. The compounds of the invention can be tested on the growth and survival of cultured T cell lines derived from patients with adult T cell leukemia associated with HTLV-1 infection. The effects of the compounds of the invention can also be evaluated on a viral replication of HTLV-1 in infected cells. The compounds of the invention can also be examined on the ability of viral protein such as Tax 1 to activate cellular transcriptional factors such as N F-? B in cultured cells. See Lindholm and others, J. Virology 70: 2525-2532, 1996, incorporated herein by reference. The ability of the compounds of the invention to attenuate the effects produced by viral proteins such as Tax 1, to decrease viral replication and / or to inhibit the growth of T cells infected by HTLV-1 can be predicted on the clinical effect beneficial in patients suffering from HTLV-1 infection and the diseases associated with it. The compounds of the formula I and the preferred compounds of the formulas la and Ib are preferably formulated before administration. Suitable pharmaceutical formulations are prepared by known procedures using well-known and readily available ingredients. To form the compositions suitable for use in the method of the present invention, the active ingredient will usually be mixed with a vehicle, or diluted by a vehicle, or enclosed within a vehicle which may be in the form of a capsule, sac, paper or other container. When the vehicle serves as a diluent, it can be a solid, semi-solid or liquid material that acts as a vehicle, excipient or medium of the active ingredient. Thus, the compositions may take the form of tablets, pills, powders, troches, sacks, sealed capsules, elixirs, suspensions, emulsions, solutions, syrups, aerosol (as a solid or in a liquid medium), hard gelatin capsules soft and hard, suppositories, solutions sterile injectables and sterile packaged powders for oral or topical application. Some examples of suitable carriers, excipients and diluents include lactose, dextrose, sucrose, sorbitol, mannitol, starches, acacia gum, calcium phosphates, alginate, tragacanth, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water syrup , methyl cellulose, methyl and propyl benzoates, talc, magnesium stearate and mineral oil. The formulations may additionally include lubricating agents, wetting agents, emulsifying and suspending agents, preservatives, sweetening agents or flavoring agents. The compositions of the invention can be formulated so as to provide rapid, sustained or delayed release of the active ingredient after administration to the patient. The compositions are preferably formulated in a unit dosage form, each dose containing from about 0.05 mg to about 3 mg, more usually about 750 mg of the active ingredient. However, it will be understood that the therapeutic dose administered will be determined by the physician in view of the relevant circumstances which include the severity of the condition to be treated, the choice of compound to be administered and the chosen route of administration. Therefore, the above dose varies and is not intended to limit the scope of the invention in any way. The term "unit dose form" refers to physically described units suitable as unit doses for human subjects and other mammals, each unit containing a predetermined quantity of active material calculated to produce the desired therapeutic effect, in association with a suitable pharmaceutical vehicle. In addition to the above formulations, most of which can be administered orally, the compounds used in the method of the present invention will also be administered topically. Topical formulations include ointments, creams and gels. Ointments are generally prepared using (1) an oil base, i.e., one consisting of fixed oils or hydrocarbons, such as white petrolatum or mineral oil, or (2) an absorbent base, i.e., one consisting of substance or Anhydrous substances that can absorb water, for example anhydrous lanolin. Commonly, after the formation of the base, either oil or absorbent, the active ingredient (compound) is added to an amount that gives the desired concentration. The creams are oily / aqueous emulsions. They consist of an oil phase (internal phase), usually comprising fixed oils, hydrocarbons and the like, such as waxes, petrolatum, mineral oil and the like and an aqueous phase (continuous phase), which comprises water and water-soluble substances, such as salts added. The two phases are stabilized using an emulsifying agent, for example, an active surface agent, such as sodium lauryl sulfate, hydrophilic colloids, such as acacia colloidal clays, "veegum" and the like. When formulating the emulsion, the active ingredient (compound) is commonly added in an amount to achieve the desired concentration.

The gels comprise a base selected from an oleaginous base, water or an emulsion-suspension base. To the base is added a gelling agent that forms a matrix in the base, increasing its viscosity. Examples of gelling agents are hydroxypropyl cellulose, acrylic acid polymers and the like. Commonly, the active ingredient (compounds) is added to the formulation at the desired concentration at a point preceding the addition of the gelling agent. The amount of compound incorporated in a topical formulation is not critical; the concentration should be within a sufficient scale to allow easy application of the formulation to the area of affected tissue in an amount that will deliver the desired amount of the compound to the desired treatment site. The common amount of a topical formulation that will be applied to affected tissue will depend on the concentration of the compound in the formulation. Generally, the formulation will be applied to the affected tissue in an amount that gives from about 1 to about 500 μg of compound per cm2 of an affected tissue. Preferably, the applied amount of the compound will vary from about 30 to about 300 μg / cm2, more preferably from about 50 to about 200 μg / cm2 and more preferably from about 60 to about 100 μg / cm2. The following formulation examples are illustrative only and are not intended to limit the scope of the invention in any way.

Formulation 1 Hard gelatine capsules are prepared using the following ingredients: Amount (mg / capsule) Active Agent 5 Starch, dry 200 Magnesium stearate 10 Total 21 5 mg The above ingredients are mixed and filled into hard gelatin capsules in amounts of 460 mg.

Formulation 2 A tablet is prepared using the following ingredients: Amount (mg / capsule) Active Agent 1 5 Cellulose, microcrystalline 10 Silicone dioxide, smoked 1 0 Stearic acid 5 Total 40 mg The components are mixed and compressed to form tablets each weighing 665 mg.

Formulation 3 Tablets are formed each containing 60 mg of active ingredient Quantity (mg / capsule) Active Agent 60 mg Starch 45 mg Microcrystalline cellulose 35 mg Polyvinylpyrrolidone (as a 10% solution in water) 4 mg Sodium carboxymethyl starch 4.5 mg Stearate of magnesium 0.5 mg Talcum 1 mg Total 1 50 mg The active ingredient, starch and cellulose are passed through a U.S. mesh screen. No. 45 and they mix thoroughly. The polyvinylpyrrolidone solution is mixed with the resulting powders which are then passed through a No. 14 mesh U.S sieve. The granules thus produced are dried at 50 ° C and passed through a U-sieve. S No. 18 mesh. Sodium carboxymethyl starch, magnesium stearate and talcum, previously passed through a No. 60 mesh US sieve, is then added to the granules which, after mixing, are compressed to a tableting machine for tablets weighing each 1 50 mg.

The principles, preferred embodiments and modes of operation of the present invention have been described in the above specification. The invention which is intended to be protected in the present, however, should not be construed as limited in the particular forms described, since it should be considered as illustrative rather than restrictive. Variations and changes can be made by those skilled in the art without departing from the spirit of the invention.

Claims (27)

1. REVIVAL DICATIONS 1. A method for inhibiting replication of human T cell lymphotropic virus type 1 in an infected cell comprising contacting the cell with an amount of inhibition of viral replication of a β-isozyme inhibitor of protein kinase C.
2. 2. The The method of claim 1, wherein the inhibitor of the protein isozyme β of protein kinase c is a bis-indolylmaleimide or a macrocyclic bis-indolylmaleimide.
3. 3. The method of claim 1 wherein the inhibitor is isozyme-selective and wherein the isozyme selectivity is selected from the group consisting of beta-1 and beta-2 isozymes.
4. 4. The method of claim 3, wherein the protein kinase C inhibitor has the following formula: wherein: W is -O-, -S-, -SO-, -S02-, -CO-, C2-C3 alkylene, substituted alkylene, C2-C6 alkenylene, -aryl, -aryl (CH2) mO- , -heterocycle-, heterocycle- (CH2) m-, bicyclic fused-, -bicyclic fused- (CH2) mO, -NR3-, -ÑOR3-, -CON H-, or -N HCO-; X and Y are independently C1-C4 alkylene, substituted alkylene, or together X, Y, and W combine to form - (CH2) n-AA-; R 1 are hydrogen or up to four optional substituents independently selected from halo, C 1 -C 4 alkyl, hydroxy, C 1 -C 4 alkoxy, haloalkyl, nitro, -N R 4 R 5, or -N HCO (C 1 -C 4 alkyl); R2 is hydrogen, CH3CO-, -N H2 or hydroxy; R3 is hydrogen, - (CH2) maryl, C1-C4 alkyl, -COO (C1-C4 alkyl), -CON R4R5, - (C = NH) NH2, -SO (C4-C4 alkyl), - SO2 (NR4R5) or -SO2 (C1-C4 alkyl); R4 and R5 are independently hydrogen, d-C4 alkyl, phenyl, benzyl or is combined with the nitrogen which is attached to form a saturated or unsaturated 5 or 6 membered ring; AA is an amino acid residue; m is independently 0, 1, 2, or 3; and n is independently 2, 3, 4, or 5, or a pharmaceutically acceptable salt, prodrug or ester thereof. The method of claim 4, wherein the protein kinase C inhibitor has the following formula. wherein Z is - (CH2) P- or - (CH2) p-0- (CH2) p-; R 4 is hydroxy, -SH, C 1 -C 4 alkyl, (CH 2) maryl, -N H (aryl), -N (CH 3) (CF 3) -N H (CF 3), or N R
5. 5 R 6; R5 is hydrogen or alkyl of ^ -04; R 6 is hydrogen, C 1 -C 4 alkyl or benzyl; p is 0, 1 or 2; and m is independently 2 or 3, or a pharmaceutically acceptable salt, prodrug or ester thereof.
6. 6. The method of claim 4, wherein the protein kinase C inhibitor has the following formula: H wherein Z is - (CH2) P-; R 4 is -NRSR 6, -N H (CF) or -N (CH 3) (CF), R 5 and R 6 are independently H or C 1 -C 4 alkyl; p is 0, 1 or 2; and m is independently 2 or 3, or a pharmaceutically acceptable salt, prodrug or ester thereof.
7. The method of claim 4, wherein the protein kinase C inhibitor comprises (S) -3,4- [N, N'-1, 1 '- ((2"-ethoxy) -3"' (O ) -4"'- (N, N-dimethylamino) -butane) -bis- (3,3'-indolyl)] - 1 (H) -pyrrol-2,5-dione or its pharmaceutically acceptable acid salt.
8. 8. A method of claim 7, wherein the pharmaceutically acceptable acid salt is selected from the hydrochloride salt and the mesylate salt
9. 9. A method for inhibiting the effect of the viral protein Tax 1 on a cell comprising putting in contacting the cell with an amount that inhibits the viral protein activity of the β-isozyme of protein kinase C.
10. 10. The method of claim 9, wherein the inhibitor of the β-isozyme of protein kinase C is a bis-indolylmaleimide or a macrocyclic bis-indolylmaleimide: 1.
11. The method of claim 9, wherein the inhibitor is isozyme-selective and wherein the isozyme selectivity is selected from the group consisting of beta-1 and beta isozymes.
12. The method of claim 1, wherein the protein kinase C inhibitor has the following formula: wherein: W is -O-, -S-, -SO-, -S02-, -CO-, C2-C6 alkylene, substituted alkylene, C2-C6 alkenylene, -aryl, -aryl (CH2) mO- , -heterocycle-, heterocycle- (CH2) m-, fused bicyclic-, -bicyclic fused- (CH2) mO, -N R3-, -ÑOR3-, -CON H-, or -N HCO-; X and Y are independently C1-C4 alkylene, substituted alkylene, or together X, Y, and W combine to form - (CH2) n-AA-; R1 are hydrogen or up to four optional substituents independently selected from halo, C1-C4 alkyl, hydroxy, C1-C4 alkoxy, haloalkion, nitro, -NR R5, or -N HCO (C1-C4 alkyl); R2 is hydrogen, CH3CO-, -NH2 or hydroxy; R3 is hydrogen, - (CH2) maryl, C1-C4 alkyl, -COO (d-C4 alkyl), -CONR4R5, - (C = NH) N H2, -SO (C1-C4 alkyl), -SO2 (NR4R5) or -SO2 (dd alkyl); R 4 and R 5 are independently hydrogen, C 1 -C 4 alkyl, phenyl, benzyl or is combined with nitrogen which are attached to form a saturated or unsaturated 5 or 6 membered ring; AA is an amino acid residue; m is independently 0, 1, 2, or 3; and n is independently 2, 3, 4, or 5, or a pharmaceutically acceptable salt, prodrug or ester thereof.
13. 13. A method of claim 12, wherein the protein kinase C inhibitor has the following formula: wherein Z is - (CH2) P- or - (CH2) p-0- (CH2) p-; R 4 is hydroxy, -SH, C 1 -C 4 alkyl, (CH 2) maryl, -NH (aryl), -N (CH 3) (CF 3) -NH (CF 3), or NR 5 R 6; R5 is hydrogen or C, -C4 alkyl; R6 is hydrogen, d-d alkyl or benzyl; p is 0, 1 or 2; and m is independently 2 or 3, or a pharmaceutically acceptable salt, prodrug or ester thereof.
14. 14. A method of claim 12, wherein the protein kinase C inhibitor has the following formula: wherein Z is - (CH2) P-; R 4 is -N R 5 R 6, -N H (CF) or -N (CH 3) (CF), R 5 and R 6 are independently H or C 1 -C 4 alkyl; p is 0, 1 or 2; and m is independently 2 or 3, or a pharmaceutically acceptable salt, prodrug or ester thereof.
15. 15. The method of claim 12, wherein the protein kinase C inhibitor comprises (S) -3,4- [N, N'-1, 1 '- ((2"-ethoxy) -3'" ( 0) -4 '"- (N, N-dimethylamino) -butane) -bis- (3,3'-indoIyl)] - 1 (H) -pyrrole-2,5-dione or its pharmaceutically acceptable acid salt.
16. A method of claim 15, wherein the pharmaceutically acceptable acid salt is selected from the hydrochloride salt and the mesylate salt
17. 17. A method for treating a mammal infected with human T cell lymphotropic virus type 1, which comprises administering to the mammal a therapeutically effective amount of an inhibitor of the β-isozyme of protein kinase C.
18. 18. The method of claim 17, wherein the inhibitor of the protein isozyme β of protein kinase C is a bis-indolylmaleimide or a macrocyclic bis-indolylmaleimide.
19. The method of claim 17, wherein the inhibitor is isozyme-selective and wherein the isozyme selectivity is selected from the group consisting of beta-1 and beta-2 isozymes.
20. The method of claim 19, wherein the protein kinase C inhibitor has the following formula: wherein: W is -O-, -S-, -SO-, -S02-, -CO-, C2-C6 alkylene, substituted alkylene, C2-C6 alkenylene, -aryl, -aryl (CH2) mO- , -heterocycle-, heterocycle- (CH2) m-, bicyclic fused-, -cyclic fused- (CH2) mO, -N R3-, -ÑOR3-, -CON H-, or -N HCO-; X and Y are independently C1-C4 alkylene, substituted alkylene, or together X, Y, and W combine to form - (CH2) n-AA-; R 1 are hydrogen or up to four optional substituents independently selected from halo, C 1 -C 4 alkyl, hydroxy, d-C 4 alkoxy, haloalkyl, nitro, -N R 4 R 5, or -N HCO (C 1 -C 4 alkyl); R2 is hydrogen, CH3CO-, -NH2 or hydroxy; R3 is hydrogen, - (CH2) maryl, d-C4 alkyl, -COO (C-> -d alkyl), -CONR4R5, - (C = NH) N H2, -SO (C? -C4 alkyl) ), -SO2 (NR4R5) or -SO2 (d-C4 alkyl); R4 and R5 are independently hydrogen, d-C4 alkyl, phenyl, benzyl or is combined with the nitrogen which is attached to form a saturated or unsaturated 5 or 6 membered ring; AA is an amino acid residue; m is independently 0, 1, 2, or 3; and n is independently 2, 3, 4, or 5, or a pharmaceutically acceptable salt, prodrug or ester thereof. twenty-one .
21. The method of claim 20, wherein the protein kinase C inhibitor has the following formula: H wherein Z is - (CH2) P- or - (CH2) p-0- (CH2) p-; R 4 is hydroxy, -SH, d-C 4 alkyl, (CH 2) mario, -N H (aryl), -N (CH 3) (CF 3) -N H (CF 3), or NR 5 R 6; R5 is hydrogen or d-d alkyl; R6 is hydrogen, C1-C4 alkyl or benzyl; p is 0, 1 or 2; and m is independently 2 or 3, or a pharmaceutically acceptable salt, prodrug or ester thereof.
22. 22. The method of claim 20, wherein the protein kinase C inhibitor has the following formula: wherein Z is - (CH2) P-; R 4 is -N RSR 6, -N H (CF) or -N (CH 3) (CF), R 5 and R 6 are independently H or d-d alkyl; p is 0, 1 or 2; and m is independently 2 or 3, or a pharmaceutically acceptable salt, prodrug or ester thereof.
23. 23. The method of claim 20, wherein the protein kinase C inhibitor comprises (S) -3,4- [N, N, -1, 1, - ((2"-ethoxy) -3 '" (0 ) -4 '"- (N, N-dimethylamino) -butane) -bis- (3,3'-indolyl)] - 1 (H) -pyrrole-2,5-dione or its pharmaceutically acceptable acid salt.
24. A method of claim 23, wherein the pharmaceutically acceptable acid salt is selected from the hydrochloride salt and the mesylate salt
25. 25. A method for treating adult T cell leukemia associated with the infection of human T cell lymphotrophic virus. Type 1, which comprises administering to an adult in need of such treatment a therapeutically effective amount of a β-isozyme inhibitor of protein kinase C.
26. 26. A method for treating a neurodegenerative disorder associated with the infection of T-cell lymphotropic virus human type 1, which comprises administering to a mammal in need of such treatment a therapeutically effective amount of an inhibitor of the β-isozyme of protein kinase C.
27. 27. The method of claim 26, wherein the chronic neurodegenerative disorder is selected from the group consisting of tropical spastic parapsis and myelopathy.