MXPA98009014A - Inhibitors of protein cinasa c, halo-sustitui - Google Patents

Abstract

The present invention relates to novel halo-substituted halo-indolmaleimide compounds of the formula. The invention further provides a method of preparing the exposed compounds and preparing the pharmaceutical formulation for use of the inhibition of Protein Kinase C in mammals.

Description

Inhibitors of Protein Kinase C, Halo-Substituted Protein kinase C (PCC) consists of a family of closely related enzymes that function as serine / threonine kinases. Protein kinase C plays an important role in signaling, gene expression and in cell differentiation and growth. At present, there are currently at least ten known isozymes of the PCC that differ in their tissue distribution, enzymatic specificity, and regulation. Nishizuka Y. Annu. Rev. Biochem. 5_a: 31-44 (1989); Nishizuka Y. Science 258: 607-614 (1992).

Isozymes of protein kinase C are chains of simple polypeptides that are in the range of 592 to 737 amino acids in length. The isozymes contain a regulatory domain and a catalytic domain connected by a linker peptide. The regulatory and catalytic domains can further be subdivided into constant and variable regions. The catalytic domain of protein kinase C is very similar to that observed in other protein kinases while the regulatory domain is unique to isozymes of PCC. The PCC isozymes demonstrate between 40-80% homology at the amino acid level between the group. However, the homology of a simple isozyme between different species is in REF .: 028760 overall greater than 97%.

Protein kinase C is a membrane-associated enzyme that is allosterically regulated by a number of factors, including membrane phospholipids, calcium and certain membrane lipids such as diacylglycerols that are released in response to the activities of phospholipases. Bell, R.M. and Burns, D.J., J. Biol. Chem. 266: 4661-4664 (1991); Nishizuka Y. Science 258: 607-614 (1992). Isozymes of protein kinase C, alpha, beta-1, beta-2 and gamma require the membrane phospholipid, calcium and diacylglycerol / phorbol esters for total activation. The delta, epsilon, eta and theta forms of the PCC are independent of calcium and its activation form. The zeta and lambda forms of PCC are independent of calcium and diacylglycerol and are thought to require only the membrane phospholipid for activation.

Only one or two isozymes of protein kinase C could be involved in a given disease state. For example, high blood glucose levels found in diabetes lead to a specific elevation of the beta-2 isozyme in vascular tissues. Inoguchi et al., Proc. Nati Acad. Sci. USA 89: 11059-11065 (1992). An elevation linked to diabetes of the beta isozyme in human platelets has been correlated with its altered response to agonists. Bastyr III, E.J. and Lu, J. Diabetes 42: (Suppl 1) 97A (1993). The human vitamin D receptor has been shown to be selectively phosphorylated by protein kinase C beta. This phosphorylation has been linked to alterations in the functioning of the receptor. Hsieh et al., Proc. Nati Acad. Sci-USA 88: 9315-9319 (1991); Hsieh et al., J. Biol. Chem. 268: 15118-15126 (1993). In addition, recent work has shown that the beta-2 isozyme is responsible for the cellular proliferation of erythroleukemia while the alpha isozyme is involved in megakaryocyte differentiation in these same cells. Murray et al. , J. Biol-Chem. 268: 15847-15853 (1993).

The ubiquitous nature of cmase C protein isozymes and their important roles in physiology provide inventives for producing highly selective PCC inhibitors. Given the evidence demonstrating the binding of certain isozymes to disease states, it is reasonable to assume that inhibitory compounds that are selective to other isozymes of PCC and other protein kinases are superior therapeutic agents. Such compounds should demonstrate greater efficiency and lower toxicity by virtue of their specificity.

Staurosporine, a microbial indolecarbazole, is a potent inhibitor of protein kinase C that interacts with the catalytic domain of the enzyme. Tamaoki et al., Biochem.

Biophys. Res. Commun. 135: 397-402 (1986); Gross et al., Biochem. Pharmacol. 40: 343-350 (1990). However, the therapeutic utility of this molecule and closely related compounds is limited by the lack of specificity for protein kinase C over other protein kinases. Ruegg, U.T. and Burgees, GM., Trends Pharmacol. Sci. 10 .: 218-220 (1989). This lack of selectivity results in unacceptable toxicity in this class of molecules.

An additional class of compounds related to staurosporine, bisindolmaleimides, has been the focus of recent work. Davis et al., FEBS Lett. 259: 61-63 (1989); Twoeny et al., Biochem. Biophys. Res. Commun. 171: 1087-1092 (1990); Toullec et al., J. Biol. Chem. 266: 15771-15781 (1991); Davis et al., J. Med. Chem. 35: 994-1001 (1992); Bitetal., J.Med. Chem. 36: 21-29 (1993). Some of these compounds have shown selectivity for protein kinase C over other protein kinases.

Although compounds demonstrating specificity to protein kinase C have been discovered, very little is known with respect to isozyme selectivity. For example, the analysis of the isozyme selectivity of staurosporine shows small isozyme selectivity with the exception of poor inhibition of the zeta isozyme relative to other enzymes. 50 McGlynn et al., J. Cell. Biochem. 49: 239-250 (1992); Ward, N.E., and O'Brian, C.A., Malee. Pharmacol. 41: 387-392 (1992). Studies of the PCC selective compound, 3- [1- (3-dimethylaminopropyl) -indol-3-yl] -4- (1H-indol-3-yl) -1H-pyrrole-2, 5-dione, suggest a slight selectivity for calcium-dependent isozymes. Toullec et al., J. Biol. Chem. 266: 15771-15781 (1991). Subsequent studies of this compound showed no difference, or possibly slight selectivity, for alpha isozymes on beta-1 and beta-2. Martiny-Baron et al., J. Biol. Chem. 268: 9194-9197 (1993); Wilkinson, et al., Biochem J. 294: 335-337 (1993).

Therefore, despite years of research and identification of classes of compounds that inhibit protein kinase C against other protein kinases, a need remains for therapeutically effective selective isozyme inhibitors. Selective inhibitors of the isozyme have utility in the treatment of conditions associated with diabetes mellitus and its complications, in addition to the treatment of ischemia, inflammation, disorders of the central nervous system, cardiovascular disease, dermatological disease and cancer.

One embodiment of the present invention are the PCC inhibitors of the formula I: where; R1 is independently hydrogen, halo, hydroxy, CX-C4 alkyl, Cj_-C4 alkoxy, NR3R4 or -NHCO (Ci-C alkyl; T is alkylene optionally substituted with halo or C 1 -C 4 alkyl; W is C 1 -C alkylene optionally substituted with halo or C 4 -C 4 alkyl; J is -X- C- or when T and W are both methylene, J is selected from the group consisting of wherein n and m are independently 1 or 2; X is oxygen, sulfur or a bond between the carbon atoms joined by the X bridge; Y is halo, alkyl Cj.-C.-i or hydrogen; RL is hydrogen or C ^ Cj alkyl, - S is -CHO or the group wherein M is hydrogen, -CH2OR5, -CH2NR3R4 or -NR3R4; R2 is hydrogen or halo; Z is hydrogen or -0R6; wherein R 3 and R 4 are independently hydrogen, C 1 -C 4 alkyl, halo (C 1 -C 4 alkyl), C 4 alkanoyl, halo (C 4 -C 4 alkanoyl) or taken together with the N atom to which they are attached form a ring of 5 or 6 members; Y R 5 and Rs are independently hydrogen, C 1 -C 4 alkyl, halo (C 3 -C alkyl, C 1 -C 8 alkanoyl halo (C 3 -C 8 alkanoyl or together form a divalent group selected from the group consisting of -CR 7 R 8- wherein R7 and R3 are independently hydrogen, alkyl dd or halo (CX-C4 alkyl) or R7 and Rβ taken together with the C atom to which they are attached form a 5 or 6 membered ring with the proviso that at least one of Y, S, T or W is halo or a substituted halo group, or T and W are both methylene.

According to another embodiment of the present invention the novel intermediates of Formula II are also provided: where R 'is independently hydrogen, halo, hydroxy, alkyl; V is oxygen or CH3 I -N-; T is C 1 -C 1 alkylene, optionally substituted with halo or C 1 -C alkyl; is Cj.-C4 alkylene optionally substituted with halo or Ci-C alkyl; J is Y I -x-c- I s or when T and W are both methylene, J is selected from the group consisting of wherein n and m are independently 1 or 2; X is oxygen, sulfur or a bond between the carbon atom bound by the X-bridge Y is halo, CX-C4 alkyl or hydrogen; S is -CHO or the group wherein M is hydrogen, -CH2OR5, -CH2NR3R4 or -NR3R4; R2 is hydrogen or halo; Z is hydrogen or -0R6; wherein R3 and R4 are independently hydrogen, C4-C4 alkyl, halo (Ct-C4 alkyl), alkanoyl-d, halo (C-J alkanoyl or taken together with the N atom to which they are attached form a ring of 5 or 6 members, and R5 and R6 are independently hydrogen, Cj.-C4 alkyl, halo (alkyl 03.-04), C ..-C4 alkanoyl, halo (Ci-C alkanoyl or together form a divalent group selected from the group consists of -CR7Ra- wherein R7 and R1¡ are independently hydrogen, Cj.-C4 alkyl or halo (C1.-C4 alkyl) or R7 and Rs taken together with the C atom to which they are attached form a ring of 5 or 6 members with the proviso that at least one of Y, S, T or W is halo or a substituted halo group, or T and W are both methylene.

• One aspect of the present invention is a method or preparation of compounds of Formula I comprising the steps of combining at a concentration of about 0.001 molar to about 1.5 molar of a compound of the formula: • where V is oxygen or NCH3, R 'is independently hydrogen, halo, hydroxy, CX-C4 alkyl, d-C * alkoxy, NR3R4 or -NHCO (Ci-C alkyl; an alkylating agent at a concentration of about 0.001 to about 1.5 molar of the formula: L L \ / T W \ ^ where L is a leaving group; T is d-C3 alkylene optionally substituted with halo or CL-C4 alkyl; W is Cj.-C4 alkylene optionally substituted with halo or Cj.-C4 alkyl; J is -X-C- I s or when T and W are methylene arabines, J is selected from the group consisting of wherein n and m are independently 1 or 2; X is oxygen, sulfur or a bond between the carbon atom bound by the X-bridge Y is halo, CX-C4 alkyl or hydrogen; S is -CHO or the group wherein M is hydrogen, -CH2OR5, -CH2NR3R4 or -NR3R4; R2 is hydrogen or halo; Y Z is hydrogen or -0RS; wherein R3 and R4 are independently hydrogen, CX-C4 alkyl, halo (Cj.-C4 alkyl), CX-C4 alkanoyl, halo (Cx-C4 alkanoyl) or taken together with the N atom to which they are attached form a ring of 5 or 6 members; Y Rs and R6 are independently hydrogen, CX-C4 alkyl, halo (CX-C4 alkyl), CX-C4 alkanoyl, halo (C1.-C4 alkanoyl) or together form a divalent group selected from the group consisting of -CR7R8- wherein R7 is hydrogen or methyl and R8 is CX-C4 alkyl or halo (CX-C4 alkyl) or taken together with the C atom to which they are attached they form a 5- or 6-membered ring, and about 0.5 to about 10 equivalents of Cs2CO3 at a rate of about 0.1 mL / hour to about 2.0 mL / hour in a polar aprotic solvent.

According to one embodiment of the present invention, a compound of formula III is provided: where; R x is C 1 -C 4 alkyl or hydrogen; T is C2-C4 alkylene optionally substituted with halo or CX-C4 alkyl; W is ethylene optionally substituted with halo or C-C alkyl • X is oxygen, sulfur or a bond between the carbon atom bound by the bridge X; Y is halo, CX-C4 alkyl or hydrogen; S is -CHO or the group wherein M is hydrogen, -CH2OR5, -CH2NR3R4 or -NR3R4; R2 is hydrogen or halo; Y Z is hydrogen or -0R6; wherein R3 and R4 are independently hydrogen, CX-C4 alkyl, halo (CX-C4 alkyl), CX-C4 alkanoyl, halo (CX-C4 alkanoyl) or taken together with the N atom to which they are attached form a ring of 5 or 6 members; Y R5 and Rs are independently hydrogen, CX-C4 alkyl, halo (CX-C4 alkyl), CX-C4 alkanoyl, halo (CX-C4 alkanoyl) or together form a divalent group selected from the group consisting of -CR7R8- wherein R7 is hydrogen or methyl and R8 is CX-C4 alkyl or halo (CX-C4 alkyl) or taken together with the C atom to which they are attached they form a 5 or 6 membered ring, with the proviso that at least one of Y, S, T or W is halo or a substituted halo group.

In another embodiment of the present invention there is provided a compound of Formula IV: where J is selected from the group consisting of wherein n and m are independently 1 or 2; Y R3 and R4 are independently hydrogen, CX-C4 alkyl, halo (CX-C4 alkyl), CX-C4 alkanoyl, halo (Cx-C4 alkanoyl) or taken together with the N atom to which they are attached form a ring of 5 or 6 members.

According to one embodiment of the present invention, a method of inhibiting the activity of PCC is provided. The method comprises administering to a mammal in need of such treatment, a pharmaceutically effective amount of a compound of Formula I. The present invention also relates to a method of selectively inhibiting the isozymes of protein kinase C beta-1 beta-2, the method comprising administering to a mammal in need of such treatment a pharmaceutically effective amount of a compound of the Formula I.

The invention also provides methods for the treatment of conditions that protein kinase C has been demonstrated to have a role in the pathology, such as ischemia, inflammation, central nervous system disorders, cardiovascular diseases, dermatological diseases and cancer. The method comprises administering to a mammal in need of treatment a pharmaceutically effective amount of a compound of Formula I.

This invention is particularly useful in the treatment of diabetic complications. Therefore, the present invention also provides a method for the treatment of diabetes mellitus, which comprises administering to a mammal in need of such treatment a pharmaceutically effective amount of a compound of Formula I.

In another aspect of this invention is a pharmaceutical formulation comprising a compound of Formula I together with one or more pharmaceutically acceptable excipients, carriers or diluents.

Detailed description of the invention As noted above, the invention provides compounds of Formula I that selectively inhibit protein kinase C. The preferred compounds of this invention are those of Formula I wherein the -TJW radicals contain from 4 to 8 atoms, which could be substituted or unsubstituted (with halo or alkyl groups). More preferably, the radicals -T-J-W contain 6 atoms. Other preferred compounds of this invention are those of Formula I wherein Rx is hydrogen, and J is And I • o-c- I s A preferred group of the compounds are the compounds of Formula V: where; R x is C 1 -C 4 alkyl or hydrogen; T is C2-C4 alkylene optionally substituted with halo or CX-C4 alkyl; W is ethylene optionally substituted with halo or alkyl Cx-C4 X is oxygen, sulfur or a bond between the carbon atom bound by the bridge X; And it is halo, CX-C4 alkyl or hydrogenated M is hydrogen -CH2OR5, -CH2NR3R4 or -NR3R4 R2 is hydrogen or halo; Z is hydrogen or -0RS; wherein R3 and R4 are independently hydrogen, CX-C4 alkyl, halo (CX-C4 alkyl), CX-C4 alkanoyl, halo (CX-C4 alkanoyl) or taken together with the N atom to which they are attached form a ring of 5 or 6 members; Y R5 and R6 are independently hydrogen, C3-C4 alkyl, halo (CX-C4 alkyl), CX-C4 alkanoyl, halo (CX-C4 alkanoyl) or together form a divalent group selected from the group consisting of -CR7R¡¡- wherein R7 is hydrogen or methyl and Rβ is CX-C4 alkyl or halo (CX-C4 alkyl) or taken together with the C atom to which they are attached form a 5 or 6 membered ring, and wherein at least one of Y, R2, Z, M, T or W is halo or a substituted halo group, more preferably, fluorine or fluorine substituted group.

Preferred compounds of Formula V include compounds wherein X is oxygen, Rx is hydrogen, T is C2-C3 alkylene, optionally substituted, and W is ethylene, optionally substituted. Preferably T and / or W are substituted with one or more fluorine groups.

In another embodiment of the present invention there is provided a compound of Formula VI: where J is selected from the group consisting of wherein Rx is alkyl or hydrogen; n and m are independently 1 or 2, - and R3 and R4 are independently hydrogen, CX-C4 alkyl, halo (CX-C4 alkyl), CX-C4 alkanoyl, halo (Cx-C4 alkanoyl) or taken together with the N atom to which they are attached form a ring of 5 or 6 members .

The term "halo" represents fluorine, chlorine, bromine or iodine.

The term "C 1 -C 4 alkyl" refers to a ring, straight or branched chain alkyl group, having one to four carbon atoms such as methyl, ethyl, n-propyl, isopropyl, cyclobutyl, n-butyl, isobutyl , sec-butyl, t-butyl and the like. Similarly, a "C2-C4 alkyl" represents a cyclo, straight or branched chain alkyl group, having one to four carbon atoms. A haloalkyl group is an alkyl group substituted with one or more halo atoms, preferably one to three halo atoms. An example of the haloalkyl group is trifluoromethyl. A Cx-C4 alkoxy is a CX-C4 alkyl group covalently linked via the -0- linkage.

The term "Cx-C4 alkylene" is a straight-chain alkylene radical of the formula - (CH2) -, wherein r is one to four. Examples of Cx-C4 alkylene include methylene, ethylene, trimethylene, tetramethylene, and the like. Similarly, "a C2-C4 alkylene" represents a straight chain alkylene of two to four carbons.

The term "Cx-C4 alkylene" represents a straight chain hydrocarbon of one to four carbons containing one or more double bonds, typically one or two double bonds. Examples of C2-C4 alkenylene groups include ethenylene, propenylene, and 1,3-butadieneyl.

The term "Cx-C4 alkanoyl" is the acyl residue of a Cx-C4 carboxylic acid. Examples of C 1 -C 4 alkanoyl groups are acetyl, propanoyl, butanoyl and the like. A haloalkanoyl group is an alkanoyl group substituted with one or more halo atoms, typically one or three halo atoms. An example of a haloalkanoyl group is trifluoroacetyl.

The term "leaving group" as used in the specification is understood by those skilled in the art. In general, a leaving group is any group or atom that improves the electrophilicity of the atom to which it joins for displacement. Preferred leaving groups are triflate, mesylate, tosylate, imidate, chloride, bromide and iodide.

The term "carboxy protective group" (P) as used in the specification refers to one of the ester derivatives of the carboxylic acid group commonly employed to block or protect the carboxylic acid group while the reactions are carried out on the other groups functional in the compound. The carboxy-protective group species employed is not critical as long as the derived carboxylic acid is stable for the condition of the subsequent reactions and can be removed at the appropriate point without interrupting the rest of the molecule. T.W. Greene and P. Wuts, Protective Groups in Organic Synthesis, John Wiley and Sons, New York, N.Y., 1991, Chapter 5, provides a list of commonly used protective groups. See also E. Haslam, Protective Groups in Organic Chemistry, J.G.W. McOmie, Ed., Plenum Press, New York, N.Y., 1973. A related term is "protected carboxy" which refers to a carboxy group protected with a carboxy protecting group.

The term "hydroxy protecting group" (P) as used in the specification refers to one of the ether or ester derivatives of the hydroxy group commonly employed to block or protect the hydroxy group while the reactions are carried out on the functional groups in the compound. The species of the hydroxy protecting group employed is not critical as long as the derivatized hydroxy group is stable to the condition of the subsequent reactions and can be removed at the appropriate point without interrupting the rest of the molecule. T.W. Greene and P. Wuts, Protective Groups in 5 Organic Synthesis. John Wiley and Sons, New York, N.Y., 1991, Chapter 5, provides a list of commonly used protective groups. Preferred hydroxy protecting groups are tert-butyldiphenylsiloxy (TEDPS), tert-butyldimethylsilyloxy (TBDMS), triphenylmethyl (trityl), K-methoxytrityl, or an alkyl or aryl ester. A related term is "protected hydroxy" which refers to a hydroxy group protected with a hydroxy protecting group.

The term "amino protecting group" (P) as used in the specification refers to substituents of the amino group commonly used to block or protect the amino functionality while reacting other functional groups in the compound. The species of the protecting group used is not critical as long as the derivative amino group is stable to the condition of the subsequent reactions and can stir at the appropriate point without interrupting the rest of the molecule. T.W. Greene and P. Wuts, Protective Groups in Organic Synthesis. John Wiley and Sons, New York, N.Y., 1991, Chapter 7, provides a list of commonly used protective groups. See also J. W. Barton, Protective Groups in Organic Chemistry, Chapter 2. Preferred amino protecting groups are t-butoxycarbonyl, phthalide, a cyclic alkyl and benzyloxycarbonyl. The related term "protected amino" defines an amino group substituted with an amino protecting group as defined.

The term "protecting groups -NH" as used in the specification refers to the sub-class of the amino protecting groups that are commonly employed to block or protect the -NH functionality while reacting other functional groups in the compound. The species of the protecting group employed is not critical as long as the derivative amino group is stable to the condition of the subsequent reactions and can be removed at the appropriate point without interrupting the rest of the molecule. T.W. Greene and P. Wuts, Protective Groups in Organic Synthesis, John Wiley and Sons, New York, N.Y., 1991, Chapter 7, page 362-385, provides a list of commonly used protective groups. The preferred -NH protecting groups are carbamate, amide, alkyl or aryl sulfonamide. The related term "protected NH" defines a group substituted with a protecting group -NH as defined.

The term "pharmaceutically effective amount", as defined herein, represents an amount of a compound of the invention that is capable of inhibiting PCC activity in mammals. The particular dose of the compound administered according to this invention will, of course, be determined by the particular circumstances surrounding the case, including the compound administered, the route of administration, the particular condition to be treated and similar considerations. The compounds can be administered by a variety of routes including oral, rectal, transdermal, subcutaneous, topical, intravenous, intramuscular or intranasal routes. For all indications, a typical daily dose will contain from about 0.01 mg / kg to about 20 mg / kg of the active compound of this invention. The daily preferred doses will be from about 0.05 to about 10 mg / kg, ideally about 0.1 to about 5 rog / kg. however, for topical administration a typical dosage is about 500 μg of compound per cm 2 of an affected tissue. Preferably, the applied amount of compound will be in the range of about 30 to about 300 μg / cm2, more preferably, about 50 to about 200 μg / cm2, and, more preferably, about 60 to about 100 μg / cm2.

The term "treatment", as used herein, describes the management and care of a patient for the purpose of cting the disease, condition, or disorder and includes the administration of an amount of the present invention to prevent the onset of symptoms or complications, relieve symptoms or complications, or eliminate tissue, condition or disorder.

The term "isozyme selective" means the preferential inhibition of the isozyme of protein kinase C (or subgroup) on the other isozymes. For example, a compound could selectively inhibit the beta-1 or beta-2 isozymes on the alpha, gamma, delta, epsilon, zeta and eta isozymes of protein kinase C. In general, the isozyme-selective compounds demonstrate a minimum of an eight-fold differential (preferably a ten-fold differential) at the dosage required to inhibit a subtype compared to the dosage required for equal inhibition or a different subtype (eg, inhibition). of the beta-1 or beta-2 isozyme of PCC as compared to the alpha isozyme of protein kinase C) as measured in the PCC test. The compounds demonstrate this differential across the range of inhibition and are exemplified in the IC50, p. ex. , 50% inhibition. Thus, for example, a selective compound of isozyme beta-1 and beta-2 inhibits the isozymes beta-1 and beta-2 of protein kinase C at much lower concentrations with lower toxicity by virtue of its minimal inhibition of the other isozymes of the CC.

The compounds of the formula I having a basic radical, e.g. ex. NR3R4, could also be in the form of their pharmaceutically acceptable acid addition salts thereof. Acids commonly used to form such salts include inorganic acids such as hydrochloric, hydrobromic, hydroiodic, sulfuric and phosphoric acids, as well as acids such as para-toluenesulfonic, methanesulfonic, oxalic, para-bromophenylsulfonic, carbonic, citric, benzoic, acetic acid and related inorganic and organic acids. Such pharmaceutically acceptable salts include in this manner sulfate, pyrosulfate, bisulfite, phosphate, monohydrogen phosphate, dihydrogen phosphate, metaphosphate, pyrophosphate, chloride, bromide, iodide, acetate, propionate, decanoate, caprylate, acrylate, formate, isobutyrate, heptanoate, propionate, oxalate, malonate, succinate, suberate, sebacate, fumarate, maleate, 2-bu-1,4-dionate, 3-hexin-2, 5-dionate, benzoate, chlorobenzoate, hydroxybenzoate or, methoxybenzoate, phthalate, xylene sulfonate, phenylacetate, phenylpropionate, phenylbutyrate, citrate, lactate, hippurate, β-hydroxybutyrate, glycolate, maleate, tartrate, methanesulfonate, propansulfonate, naphthalene-1-sulfonate, naphthalene-2-sulfonate, mandelate and the like salts.

In addition to pharmaceutically acceptable salts, other salts according to the present invention may be used. These could serve as intermediates in the purification of the compounds, in the preparation of other salts, or in the identification and characterization of the compounds or intermediates.

The pharmaceutically acceptable salts of the compounds of Formula I may also exist as various solvates, such as with water, methanol, dimethylformamide ethyl acetate and the like. Mixtures of such solvates can also be prepared. The source of such solvates can be of the crystallization solvent, inherent in the preparation or crystallization solvent, or extrinsic to such solvents. Such solvates are within the scope of the present invention.

The various stereoisomeric forms of the compounds of Formula I may exist; for example, T or W could include a chiral carbon atom in the substituted alkylene radical. The compounds of Formula I are typically prepared as racemates and can conveniently be used as such, but the individual enantiomers can be isolated or synthesized by conventional techniques if desired. Such racemates and individual enantiomers and mixtures thereof form part of the present invention.

The present invention also encompasses pharmaceutically acceptable prodrugs of the compounds of Formula I. A prodrug is a drug that has been chemically modified or could be biologically inactive (at the site of action of the enzyme), but can be degraded or modified by one or more enzymatic processes or other processes in vivo to produce the bioactive form of origin. The prodrug preferably has a different pharmacokinetic profile than the origin, improving easier absorption through the epithelial mucosa, better salt formation or solubility, and / or improved systematic stability (for example, an increase in plasma half-life). . Conventional procedures are described for the selection and preparation of specific prodrug derivatives, (eg, in H. Bundgaard, Design of Prodrugs, (1985).) Typically, such chemical modifications include the following: 1) ester or amide derivatives which could be broken down by esterases or lipases; 2) peptides that can be recognized by specific or non-specific proteases; or 3) derivatives that accumulate in the action site through the membrane selection of a prodrug form; or a modified prodrug form, or any combination of 1 to 3, supra.

The synthesis of certain bis-indole-N-maleimide derivatives is described in Davis et al., U.S. Pat. 5,057,614, the disclosure of which is incorporated herein by reference. In general, the compounds of the present invention could be prepared as follows: Scheme 1 I 2 I The halo group according to scheme 1 is preferably fluorine, chlorine, bromine or iodine. Compound 1 is preferably 2,3-dichloro N-methylmaleimide. The reaction between Compound 1 and indole, Compound 2, is commonly known as a Grignard reaction. The reaction is carried out in an inert solvent, such as toluene, at a temperature between room temperature and the reflux temperature of the reaction mixture. More significantly, the reaction depicted in Scheme I is dependent on the solvent conditions. When carried out in a solvent system of toluene: THF: ether, the reaction provides Compound 3. greater than 80 percent yield and greater than 95 percent purity. The product is precipitated from the reaction mixture with ammonium chloride, NH 4 Cl. The resulting intermediate, Compound 2_, could be isolated by standard techniques.

Bis-3,4 (3'-indolyl) -lN-methyl-pyrrole-2, 5-dione, Compound 3_, could then be converted by alkaline hydrolysis to the corresponding anhydride of formula 4 by techniques known in the art and described in Brenner et al., Tetrahedron 44: 2887: 2892 (1988). Preferably, Compound 3_ is reacted with 5N KOH in ethanol at a temperature that is in the range of 25 ° C to the reflux temperature to produce the compound of Formula 4 The compounds of Formula 3 are generally more stable than the compounds of Formula 4. > therefore, it is preferred that Compounds 3. react in accordance with Scheme 2 to produce the compounds of the Formula. However, one skilled in the art would recognize that the compounds of Formula 4 could also be reacted according to scheme 2.

Compounds 3 or 4 \ II / T, J and W are the same as previously described. L is a leaving group such as chlorine, bromine, iodine, mesyl, tosyl and the like. L could also be a hydroxy or other precursor that could easily be converted to a good leaving group by techniques known in the art. For example, hydroxy could easily be converted to a sulfonic ester such as mesyl by reacting the hydroxy with methanesulfonyl chloride to produce the mesylate leaving group.

The reaction represented by Scheme 2 is carried out by any of the known methods of preparing N-substituted indoles. This reaction usually involves approximately equimolar amounts of the two reagents, although other relationships are operative, especially those where the alkylation reagent is in excess. The reaction is best carried out in a polar aprotic solvent employing an alkali metal salt or other such alkylation conditions as appreciated in the art. When the leaving group is bromine or chlorine, a catalytic amount of the iodide salt, such as potassium iodide, could be added to accelerate the reaction. The reaction conditions include the following: Potassium hexamethyldisilazide in dimethylformamide or tetrahydrofuran, sodium hydrate in dimethylformamide.

Preferably, the reaction is carried out under the slow reverse addition with cesium carbonate either in acetonitrile, dimethylformamide (DMF), or tetrahydrofuran (THF). The temperature of the reaction is preferably from about room temperature to about reflux temperature of the reaction mixture.

One skilled in the art would recognize that the reaction described in Scheme 2 could be employed with the compounds L-T 'and L-W', wherein T 'and W are a protected carboxy, protected hydroxy or a protected amine. After the alkylation of Scheme 2, T 'and W' could be converted to the radicals capable of coupling to form J. The coupling of T 'and W1 to form the ether or thioether derivatives is known in the art and is described in, for example Ito et al., Chem. Pharm. Bull. 41 (6): 1066-1073 (1993); Kato, et al. Synthesis 1981: 457; Harpp, et al., J. Am. Chem. Soc. 93: 2437 (1971); and Evans et al., J. Or. Chem. 50: 1830 (1985).

One skilled in the art would also recognize that compound 3 could be converted to the compounds of formula I in a two-step synthesis as described in Scheme 3.

C II T, J, W, V and L are as previously defined. L2 is a protected hydroxy or other group that could easily be converted to a good leaving group by techniques known in the art. The coupling between Compound 3 or 4 and Compound E is an alkylation as discussed previously. The monoalkylated intermediate, 7, is deprotected, and L2 is converted to a leaving group. For example, if hydroxy is protected with t-butyldimethylsilyl (TBDMS), the TBDMS is selectively removed using acid methanol. The resulting free hydroxy is then converted to a leaving group, such as an alkyl halide, preferably an alkyl iodide or bromide (CBr4 in triphenylphosphine) or sulfonate (mesyl chloride in triethylamine). The microlide is then formed by alkylation under slow inverse addition to a solution of the base, such as potassium hexamethyldisilazide, or sodium hydrate, but preferably Ca 2 CO 3 in an aprotic polar solvent such as acetonitrile, DMF, THF at the temperature in the range of the environment to reflux.

The compounds of Formula I could be prepared in substantially higher yield when the alkylation is carried out under slow inverse addition to Cs2CO3 in a polar aprotic solvent. The reverse addition involves the combination of a mixture of the compound and the alkylating agent (Scheme 2) or the compound (Scheme 3) with the base at a rate of about 0.1 mL / hour to about 2.0 mL / hour. The concentration of each reagent in the mixture is about 1.5 molar to about 0.001 molar. When carried out with the monoalkylated compound (Scheme 3) the concentration is from about 3 molar to about 0.001 molar. The slow addition results in a concentration of reactants in the reaction vessel from about 0.01 molar to 1.5 molar. One skilled in the art would recognize that it could be used at a higher rate of addition at a lower concentration of the reactants. Also, at a slower rate of addition, a higher concentration of the reactants could be used in the reaction. Preferably, the compound is added at about 0.14 mL / hour with the compound and alkylating agent at 0.37 molar. It is preferred that the Cs2C03 be in excess - more preferably a 4: 1 ratio of Cs2C03 to the alkylating agent. Preferred polar aprotic solvents are acetonitrile, dimethylformamide (DMF), acetone, dimethyl sulfoxide (DMSO), dioxane, diethylene glycol methyl ether (diglyme), tetrahydrofuran (THF), or other polar aprotic solvents in which the reactants are soluble. The reaction is carried out at temperatures in the range of about 0 ° C to reflux. One skilled in the art would recognize that the ratio of the mixture of the compound and the alkylating agent is not critical. However, it is preferred that the reagents are mixed in a ratio of 0.5 to 3 equivalents from one to the other. More preferably, the reactants are mixed 1: 1.

When V is N-CH 3, Compound II is converted to the corresponding anhydride (V is 0) by alkaline hydrolysis. Alkaline hydrolysis involves the reaction of the compound with the base (such as sodium hydroxide or potassium hydroxide), either in Cx-C4 alcohol (preferably ethanol), DMSO / water, dioxane / water, or acetonitrile / water at a temperature in the range of approximately 25 ° C to preferably approximately reflux. The concentration of the reagents is not critical.

The anhydride (V is 0) is converted to the maleimide of the Formula I by ammonolysis. The ammonolysis involves the reaction of the anhydride with an excess of hexamethyldisilazane or an ammonium salt (ammonium acetate, bromide, or chloride) and Cx-C4 alcohol (preferably methanol) in a polar aprotic solvent such as DMF at room temperature. Preferably, the hexamethyldisilazane or an ammonium salt is reacted at a ratio greater than about 5: 1 equivalents of anhydride.

Scheme 4 represents the preparation of a useful intermediate according to the present invention for the preparation of the compounds of Formula III, wherein X is 0.

Scheme 4 OEt 22 26 • 31- 26 The following examples and preparations are provided simply to further illustrate the invention and not to limit it.

Example 1 Synthesis of Bisindolylmaleimides linked by six Bisfluorinated Atoms A compound of formula VII: it occurs as follows OEt OH • Compound 8a is converted to the compound of Formula VII using the steps depicted in scheme 4.

A compound of formula VIII: It occurs as follows: This compound is converted to the compound of Formula VIII using the steps depicted in scheme 4.

A compound of formula IX: It proceeds as follows: This compound, wherein R 'is H or F, is converted to the compound of Formula IX using the steps depicted in Scheme 4.

Example 2 Synthesis of Bisindolylmaleimides linked by Seven C2-Monofluorinated Atoms A compound of the formula X: It occurs as follows: This compound is converted to the compound of Formula X using the steps depicted in Scheme 4.

Example 3 Synthesis of Bisindolylmaleimides linked by Seven C6 Monofluorinated Atoms A compound of formula XI: It occurs as follows: This compound is converted to the compound of Formula XI using the steps depicted in Scheme 4.

Example 4 Synthesis of Bisindolylmaleimides linked by Seven C6-Difluorinated Atoms A compound of formula XII: It occurs as follows: This compound is converted to the compound of Formula XII using the steps depicted in Scheme 4.

Example 5 Synthesis of Thio-linked Bisindolylmaleimides by Six Fluorinated Atoms A compound of formula XIII: It produces as follows: OEt This compound is converted to the compound of Formula XIII using the steps depicted in Scheme 4.

Example 6 Synthesis of C4 Lateral Chain Derivatives of Bisindolylmaleimides Linked by Six Fluorinated Atoms A compound of the formula XIV: It occurs as follows: This compound becomes the compound of the Formula XIV using the stages represented in the scheme.

Example 7 Synthesis of Bisindolylmaleimides Linked by Six Volumes of Fluoroalkene A compound of the formula XV: ; It occurs as follows: This compound is converted to the compound of Formula XV using the steps depicted in Scheme 4.

Example 8 Synthesis of Bisindolylmaleimides Linked by Seven Fluoroalkene Atoms A compound of formula XVI (fo 2 It occurs as follows: This compound is converted to the compound of Formula XVI using the steps depicted in Scheme 4.

A compound of Formula XVII: (fo It occurs as follows: This compound is converted to the compound of Formula XVII using the steps depicted in scheme 4. A compound of formula XVIII: XVIII (forms E and Z) It occurs as follows: This compound is converted to the compound of Formula XVIII using the steps depicted in Scheme 4.

Example 9 Synthesis of Bisindolylmaleimides Linked by Six C2-Methylated Fluorinated Atoms A Compound of Formula XIX: it occurs as follows: b. aldehyde OH s (3 -indolyl) maleimide, Cs2C03 XIX Example 10 Synthesis of Bisindolylmaleimides Linked by Six Fluorinated Atoms Derived C3 Example 11 Synthesis of Bisindolylmaleimides Linked by Six Fluorinated Atoms Derived C3 Example 12 Synthesis of Bisindolylmaleimides Linked by Six Fluorinated Atoms C3 Derivatives In the following examples and preparations, the melting point, the nuclear magnetic resonance spectrum, the mass spectrum, the high pressure liquid chromatography on silica gel, N, N-dimethylformamide , palladium on charcoal, tetrahydrofuran and ethyl acetate are abbreviated Pt. Fus, NMR, MN, HPLC, DMF, Pd / C, THF and EtOAc, respectively. The terms "NMR" and "MS" indicate that the spectrum was consistent with the desired structure.

Example 13 Preparation of bisindolylmaleimide of the formula (3S.4R) -2-fluoro-3-hydroxy-4,5-O-isopropylidene ethyl ethanoate.

To a stirred round bottom flask containing 50 mL of anhydrous THF and 3.1 mL (0.03 mol) of hexamethyl disilazane (HMDS) at 0 ° C, was added 12 mL (0.03 mol) of 2.5M solution of butyllithium in hexane. . The resulting mixture was allowed to return to ambient temperature and was stirred for 10 min. Then it was decreased to -85 ° C. A mixture of 1.80 g (0.10 mol) of hexamethyl triamide phosphate (HMPA) and 1.00 mL (0.010 mol) of ethyl fluoroacetate was added dropwise as quickly as possible while the temperature was not allowed to rise to -85. ° C. After an additional 5 min, 880 mg (0.0067 mol) of 2,3-0-isopropylidene D-glyceraldehyde (8) was added. The mixture was stirred for 10 min and then quenched at -85 ° C with 5 mL of saturated ammonium chloride. Warming to room temperature the mixture was diluted with 80 L of CH2C12 and washed with water (60 ml x 3). The CH2C12 layer was separated and dried with Na2SO4. After evaporation of the volatiles in vacuo, the residue was loaded on a short silica gel column. It was washed with CH2C12 then CH3CN / CC14 (or ethyl acetate), yielding the crude product.

The crude product was chromatographed on a column of silica gel using CH3CN / CC14 at 15% (v / v) as the eluent, yielding two fractions with partial separation of the two diastereomers: fraction 1, 177 mg, (9a, 34% , 9b, 66% based on XHRMN).

Fraction 2, 377 mg (9a, 62%, 9b, 38%). Overall performance of 34.7%, in which 9a, was 52.7%, and 9b was 47.3% 9b: 2S 10b: 2S (3S, 4R) 3-allyloxy-2-fluoro-4,5,0-isopropylidene pentanoate ethyl.

Alcohol 9, 327 mg (1.39 mmol, which is a mixture of 9a, 62% and 9b, 38%), was evaporated with toluene and 327 mg (1.39 mmol), dissolved in 6 mL of cyclohexane. Under N2 atmosphere and stirring, allyl trichloroacetimidate (423 μL, 561 mg, 2.78 mmol) was added, followed by trifluoromethanesulfonic acid (20 μL) in 5 μL portions for 20 min. A precipitate began to form instantaneously. Oily dark coffee. The reaction mixture was stirred at room temperature for 60 h. A white precipitate formed, and such a precipitate was filtered and washed twice with cyclohexane. The filtrate was evaporated to an oily liquid. Chromatograph on a silica gel column using 10% ethyl acetate-hexane (v / v) as the eluent, affording two fractions. The first fraction contained mainly 10a and the second fraction 10b. The fractions were purified on a silica column using 25% hexane / CH2Cl2 (v / v) as the eluent, yielding 10a, 133 mg (34.8%) and 10b, 83 mg (21.7%). Total yield 56.5%. 10b: 2S llb: 2R (3S.4R) 3-Allyloxy-2-fluoro-4,5-0-isopropylidene pentanoi (11).

Ester 10a, 60 mg (0.217 mmol) was evaporated twice with toluene. This was dissolved in 3.0 mL of dry THF and cooled to -75 ° C. Under N2 atmosphere with stirring, toluene solution DIBAL-H 0.68 mL (1.29 M, 0.877 mmol) was added dropwise during 20 min. The resulting solution was stirred at -75 ° C for another 1.5 h. Then it was allowed to warm to -5 ° C and quenched with 4 mL of ethyl acetate. After stirring for 10 min. Wet Na 2 SO 4 (2 g) was added. The mixture was stirred at -5 ° C for 2 h. The solid was filtered and washed twice with ethyl acetate. The filtrate was evaporated under reduced pressure. The residue was chromatographed on a column of silica gel using 30% ethyl acetate-hexane (v / v) as the eluent, yielding pure lia, 32 mg (63.4%). llb is obtained in the same way as llb.

Diol (12): Compound 11 (a mixture of lia and llb), 23 mg (0.099 mmol) was dissolved in 3 mL of CH30H and 1 mL of CH2C12. At -78 ° C, it was bubbled with 03 until the solution changed to bright blue. The solution was then bubbled with argon for 5 min. A drop of dimethyl sulfide was added, and the mixture was stirred for 10 min. Sodium borohydrate 30 mg (0.794 mmol, 8 eq) was added and stirred for 5 min. The reaction mixture was allowed to return to room temperature and then stirred for another 1 h. After adding 3 drops of saturated aqueous Na 4 Cl solution, the mixture was stirred for another 1 h. The volatiles were removed in vacuo. The residue was dissolved in methanol, and ethyl acetate was added to replace the methanol by co-evaporation. The white precipitate was filtered and the filtrate was evaporated. The residue was chromatographed on a column of silica gel using ethyl acetate. After the removal of the traces of the unknown components, 13 mg (55.3%) of 12a and 7 mg (29.8%) of 12b were obtained.

Mesilation and Coupling Reactions The diol (12a), 12 mg (0.050 mmol) was dissolved in 5 mL of anhydrous ethyl ether and cooled to 0 ° C under N2 atmosphere. 35 μL (0.250 mmol, 5 eq.) Of Et3N was added with stirring, followed by 20 μL (0.25 mmol, 5 eq.) Of mesyl chloride. The resulting mixture was stirred at 0 ° C for 5 h. The precipitate was filtered and washed with ethyl ether. The filtrate was washed with water (2x) and brine (2x) and dried with Na2SO4. After evaporation of the solvents in vacuo, a yellowish oil 13a, 9 mg (43.3%) was obtained.

The precipitate was dissolved in water and extracted with ethyl acetate. The ethyl acetate layer was washed with aqueous NaHCO 3 solution twice and dried with Na 2 SO 4. After evaporation of the solvent in vacuo, 11 mg (55.3%) of 13a was obtained. Totally 20 mg of 13a was obtained, a 100% yield.

Dimesylate 13a, 20 mg (0.0507 mmol) and bisindolyl-maleimide 17.3 mg (0.0507 mmol) were combined and dissolved in 2.5 mL of anhydrous DMF (dried with molecular sieves) and added via a syringe pump in addition to a suspension of cesium carbonate (66 mg, 0.203 mmol in 3 mL of anhydrous DMF at 50 ° C under N2 for 40 h). The reaction mixture was filtered at 50 ° C for another 10 h. Evaporated in vacuo to remove volatiles. The residues were dissolved in CHC13 and washed with saturated aqueous NaHCO3 solution and brine. The chloroform layer was separated and dried with Na 2 SO 4. After evaporation, the red solid residue was chromatographed on a column of silica gel with elution of acetone / CHCl 3 at 10% (v / v). The desired compound 14a, 7 mg (25.4%) was eluted first, followed by one of the initiator materials, bisindol-maleimide 13 mg (75%).

Example 14 Preparation of bisindolyl maleimide of the formula: The compounds 21a and 21b are stereoisomers, 21a is in the 2R conformation; 21b is in the 2S conformation. The nomenclature of the chiral center changes when the ester is reduced to alcohol (eg compound 24a becomes 2S). 22 ' To a shaken round-bottomed flask containing 40 mL of anhydrous THF and 5.0 mL (0.0255 mol) of hexamethyldisilazane 9.0 mL (0.0225 mol) of a 2.5 M butyl lithium solution in hexane was added and reacted in a low volume. ice under N2. After stirring at room temperature for 10 min, it was cooled to -78 ° C, and 3.6 g (0.020 mol) of HMPA and 2.0 mL (0.020 mol) of ethyl fluoroacetate were added dropwise in 5 min. After stirring for an additional 5 min, 760 mg (4.46 mmol) of cyclohexylidene glyceraldehyde (21, prepared as Ref. JOC, 1992, 57 page 648, and JOC, 1995, 60, pages 585-587, distilled at 60 ° C. /0.5 mmHg) was added quickly. The mixture was allowed to stir for an additional 10 min, and then quenched at -78 ° C with 5 mL of saturated ammonium chloride. Warming to room temperature the mixture was diluted with 60 mL of hexane. The hexane layer was separated. The remaining layer was extracted with 30 L of hexane. The hexane solutions were combined and washed with saturated ammonium chloride (100 mL x 3) and dried with Na 2 SO 4. After evaporation under reduced pressure, the residue was chromatographed on a column of silica gel using 30% ethyl acetate / hexane, yielding the main product 22 '740 mg (47.6%), which is a mixture of the two isomers with the major isomer 22 'a (74%) and the minor isomer 22' b (26%) based on "? RMN." The aqueous wash was extracted with CHC 13. After evaporation and chromatography, 92 mg (7.5%) ) of 21 was obtained from the chloroform layer, which is also a mixture of the two isomers.The pure 22'a isomer was obtained from the mixture 22 'by chromatography on a silica gel column eluted with CHC13. 22 'The initiator material 22', 740 mg (2.1 mmol) was dissolved in 60 mL of CH3OH. 1.2 g of citric acid was added. The mixture was stirred at room temperature for 4 h. The solvent was removed in a rotary evaporator. The residue was placed with ethyl acetate (100 mL) and washed with saturated NaHCO 3 solution (100 mL x 2) and water. The ethyl acetate layer was dried with Na 2 SO 4 and evaporated, yielding 22,600 mg (100%), which is a mixture of isomer 22a (69%) and 22b (31%) based on XHRMN. Pure 22a was obtained by 22 'hydrolysis under the same condition producing a 100% yield.

The alcohol 22, 655 mg (2.37 mmol, a mixture of isomer 21a (69%) and 21b (31%) was dissolved in 30 mL of cyclohexane and 1.50 mL (9.8 mmol) of allyl trichloroacetimidate was added. CF3SO3H was added dropwise over 30 min.The reaction mixture was stirred at room temperature under N2 for 16 h.TLC showed that there was approximately 20% of the initiator material.Additional 60 μL of CF3S03H was added and the reaction mixture was added. The mixture was stirred for 24 h, the precipitate was filtered and washed with cyclohexane, the filtrate was evaporated and the residue was chromatographed on a column of silica gel using 10% ethyl acetate as eluent, yielding 23a, 511 mg (68.1%). and 23b, 160 mg (21.3%).

Pure 23a was also obtained from pure 21a using the same condition above for comparison.

Ester 23a, 635 mg (2.00 mmol) was evaporated with toluene twice and dissolved in 10 mL of anhydrous THF. A suspension of 200 mg (5.27 mmol, 2.5 eq.) LiAlH4 in 40 mL of anhydrous THF was added dropwise at -78 ° C under N2 with stirring.

After adding the sample, the reaction mixture was stirred for 20 min. It was heated to 0 ° C and stirred at 0 ° C for 20 min. Then 5 mL of ethyl acetate was added. After stirring for 5 min. , 4 g of wet sodium sulfate was added. The mixture was stirred for 30 min. The solid was filtered and washed with ethyl acetate twice. The filtrate was evaporated in a rotary evaporator. The residue was chromatographed on a column of silica gel using 30% ethyl acetate / hexane as the eluent. After the removal of some starting material 23a, 19 mg, the main product was eluted, such as 24a, 413 mg (75.0%), followed by some 24b, 30 mg (5.5%).

Compound 24b (44 mg) was obtained by reduction of 23b (112 mg) using DIBAL-H in THF using a similar procedure as above, in 45% yield. 24a 25a The alcohol 24a, 295 mg (1.08 mmol) was dissolved in 20 mL of the solvent mixture of CH30H / CH2C12 (1: 1). It was cooled to -78 ° C. Ozone was bubbled until a blue color began to appear. Argon was then bubbled to exclude the excess of 03. Several drops of CH3SCH3 were added and the solution was stirred for 5 min. After 245 mg (6.48 mmol), NaBH4 was added at -78 ° C. After stirring for 5 min., The mixture was allowed to return to room temperature and stirred for 1 hour. The volatiles were removed in vacuo. The residue was chromatographed on a column of silica gel using ethyl acetate as the eluent, yielding 25a, 232 mg (77.5%).

Diol 25b was obtained following the. same previous procedure. The reaction was started with 230 mg of 24 b, and 158 mg of 25b was obtained with a yield of 67.7%. 25th 26th The diol 25a, 195 mg (0.70 mmol) was dissolved in 50 mL of diethyl ether. 585 μL triethyl amine (4.2 mmol) was added and followed by 342 μL (4.2 mmol) of methanesulfonyl chloride. The mixture was stirred at room temperature under N2 for 3 h. 50 mLs of water were added to dissolve the precipitate. The ether layer was separated and washed with water (50 mL x 2). After drying with anhydrous Na 2 SO 4, it was evaporated under reduced pressure to yield a yellowish liquid 26a, 308 mg (100%).

To a round bottom flask containing 768 mg (2.36 mmol) of cesium carbonate in 40 mL of anhydrous DMF at 50 ° C under N2, 10 mL of DMF solution containing 26a, 257 mg (0.59 mmol) and bisindolmaleimide, 202 mg (0.59 mmol) was added dropwise via a syringe pump over a period of 48 h. After stirring at 50 ° C for 24 h.

Further, the reaction mixture was diluted with 100 mL of CHC13 and washed with brine (50 mL x 2), then water (50 mL x 2). The chloroform layer was dried with anhydrous Na 2 SO 4 and evaporated under reduced pressure. The residue was dissolved in CHC13 and chromatographed on a silica gel column using 5% acetone / CHCl3 as the eluent. The first component eluted is the desired product 27a, 185 mg. It was recrystallized from CHC13 / CH30H, yielding 130 mg (37.7%) and a filtrate.

Compound 27b was obtained in the same manner as above, except that the addition time of mesylate and bisindolyl maleimide via the syringe pump was 80 h. The reaction was started with 97 mg diol 25b, and 64 mg of 27 b was obtained with an overall yield of 26.1%.

The initiator material 27a, 50 mg (0.099 mmol) was dissolved in 50 mL of methanol. 1 mL of water and 200 mg of p-toluenesulfonic acid monohydrate (1.05 mmol) were added. The reaction mixture was stirred at 50 ° C for 4 h. It was evaporated on a rotary evaporator and the residue was chromatographed on a silica gel column using 5% CH30H / CHC13 as the eluent. The main component 28a was recrystallized from acetone / CH3OH, yielding crystalline 28a, 23 mg and a filtrate containing 18 mg of 28a (95.0%). 28 a 29a Diol 28a, 55 mg (0.109 mmol) was dissolved in 20 mL of acetone-HOAc mixture (1: 1). To the solution, 100 mg of NAI04-3H20 (0.373 mmol) in 2.5 mL of water was added. The mixture was stirred at room temperature for 2.5 h. It was evaporated in a rotary evaporator to reduce the volume of the solution in half, and then it was diluted with 30 mL of CH2C12. The mixture was then washed with water twice, aqueous NaHC03 and again water. The CH2C12 layer was separated, dried with Na2SO4 and evaporated to dryness, yielding the crude aldehyde 29a.

The crude aldehyde 29a was dissolved in 14 mL of CH2C12 and cooled to -78 ° C. Sodium borohydrate, 30 mg (0.79 mmol) was dissolved in 6 mL of SP reagent alcohol and added to the solution. The mixture was stirred under N2 for 40 min. It was quenched with 500 μL of CH3CH0 and then allowed to return to room temperature. The mixture was evaporated to reduce the volume of the solution in half and then mixed with 10 mL of C2H5OH, 2 mL of saturated aqueous solution of sodium potassium tartrate. The mixture was stirred for 5 h. It was evaporated and placed with 30 ml of CH2C12. The CH2C12 layer was separated, washed with water three times and dried with Na2SO4. After evaporation, the residue was chromatographed on a column of silica gel using 10% ethyl acetate in CH2C12, yielding compound 30 'a, 7 mg (14.1%). The main component was eluted with ethyl acetate, yielding the desired product 30a, 41 mg (79.3%).

Compound 30b of the corresponding diol 28b was obtained (with a yield of 63.4%) following the same procedure as the previous one except that the solvent for cutting the diol was a mixture of CH3CN-H20 (2: 1).

Alcohol 30a, 16 mg (0.0338 mmol) was dissolved in 10 mL of CH2C12. 70 μL (0.52 mmol) of triethylamine and 28 μL (0.34 mmol) of methanesulfonyl chloride were added thereto. The reaction mixture was stirred at room temperature under N2 for 3 h. It was transferred to a separatory funnel and washed with water three times.

After evaporation, 31a crude, 23 mg, was produced. the TLC shows a single point.

The crude mesylate 31a, 23 mg (approximately 0.0338 mmol) was placed in a 25 mL round bottom flask and 12 mL of C2HsOH was added. The flask was cooled using a dry ice / acetone bath. In this, 1.2 g of HN (CH3) 2 and 3 mL of water via cannula. The flask was then sealed with a Teflon plug and tied with copper wire. This was stirred at 100 ° C for 8 h. After the flask was cooled to room temperature, the volatiles were removed in a rotary evaporator. The residue was dissolved in 20 mL of ethyl acetate and washed with aqueous HHCOa (20 mL x 2), water (20 mL x 2) . After removing the solvent, 24 mg of the crude product 32a was produced, which was recrystallized from methanol to obtain pure 32a.

Imide 32a, 24 mg (0.048 mmol) was dissolved in a mixture of 3 mL of C2H5OH and 3 mL of 5N KOH. It was stirred at 80 ° C for 24 h. The ethanol was stirred in a rotary evaporator and the aqueous suspension was cooled to 0 ° C and acidified with 5N HCl. A hasty violet appeared. After stirring for 10 min, the aqueous mixture was neutralized with dilute KOH and extracted with ethyl acetate. The ethyl acetate layer was washed with aqueous NaHCO 3 twice and water. After drying with K2CO3 and evaporation, 24 mg of crude 33a was produced.

. The anhydride 33a, 24 mg was dissolved in 5 mL of anhydrous DMF. To this was added 250 μL (1.19 mmol 1,1,1,3,3,3-hexamethyl disilazane, followed by the addition of 25 μL of CH30H (0.62 mmol). The resulting mixture was stirred at room temperature under N2 for 38 h. The volatiles were removed in vacuo. The residue was dissolved in 6 mL of the mixture of the 1 N CH3CN-HCl solution (2: 1) and stirred at room temperature for 1 h. The organic solvent was removed and the aqueous suspension was neutralized with IN KOH and extracted with ethyl acetate. The ethyl acetate layer was washed with KOH 0. 1 N and water twice and evaporated to dryness. The residue was separated on a column of silica gel eluted with ethyl acetate. The second eluted band is the desired product 14a which was recrystallized from CH2C12-hexane, yielding 5 mg (overall yield of 30a, 20%).

Example 15 Alternative Scheme for the Production of Compounds 34 b) from Compounds (a and b).

DMF, HMDS CH3CN, MeOH, 36 | ? N Hcl 34a + b Alcohol 30a, 50 mg (0.106 mmol) was mixed with 20 mL of 5N C2H5OH-KOH (1: 1). It was stirred at 70 ° C under N2 for 20 h. It was then cooled to 0 ° C and acidified with 5N HCl. The red precipitate appeared immediately. Methylene chloride, 40 mL was added. The organic layer was separated and washed with water (30 mL x 4), and dried with Na 2 SO 4. After evaporation, the residue was chromatographed on a column of silica gel using 5% ethyl acetate in CH2C12 to yield 35a, 33 mg (68%).

The anhydride 35a, 54 mg (0.117 mmol) was dissolved in 5 mL of anhydrous DMF. HMDS (1, 1, 1, 3,3,3, -hexamethyl disilazane). 500 μL (2.36 mmol) and methanol, 48 μl (2.36 mmol) were added. The mixture was stirred under N2 at room temperature for 36 h. The volatiles were removed in vacuo and the residue was stirred with 10 mL of CH3CN and 5 mL of IN HCl for 1 h. It was then concentrated and extracted with CH2C12. The methylene chloride layer was washed with water, brine, and dried with Na 2 SO 4, and evaporated. The residue was separated on a column of silica gel using CH2C12-CH3CN (9: 1), yielding the first component which was the initiator material at 36a, 31 mg (57.4%).

Compound 36b, 3.4 mg (39%) was obtained from the corresponding alcohol 30b, 9 mg (0.019 mmol) following the same procedure as above except that more excess HMDS (250 μL, 1.18 mmol) and methanol (24 μL) were used. , 1.18 mmol) than for 36a.

Amination: Compound 36a, 31 mg (0.068 mmol) was dissolved in 15 mL of anhydrous THF. To this were added 240 μL (1.56 mmol) of triethylamine and 84 μL (1.02 mmol) of methanesulfonyl chloride under a nitrogen atmosphere. The mixture was stirred at room temperature for 3 h. The volatiles were removed in vacuo. The residue was dissolved in 30 mL of CH2C12 and washed with IN HCl, brine twice, dried with Na2SO4 and evaporated. The residue was dissolved in 6 ml of distilled THF and 1 ml of 40% dimethylformamide in water. The flask was sealed with a Teflon cap and stirred at 50 ° for 24 h. The mixture was cooled to 0 ° C and evaporated to remove the volatiles. The residue was purified on a column of silica gel using 0-10% Et3N in ethyl acetate, yielding the desired compound 34a, 13.2 mg (40.2%).

Compound 34b (1.9 mg) was obtained from 36b (3.4 mg) using the same procedure as for 34a with a yield of 52%.

Example 16 Synthesis of a Dithiocarbamate Derivative: mg of Compound 38 (0.04 mmol, 1 eq.) Was transferred to a 25 mL round bottom flask equipped with a stir bar, partition and N2. 10 μL of THF was added via cannula followed by 4.5 mg of triethylamine (0.044 mmol, 1. 1 eq.) And then 3.8 mg of carbon disulfide (0.05 mmol, 1. 2 eq.) Via syringe. This red solution was allowed to stir for about 15 min and then 7.1 mg of methyl iodide (0.05 mmol, 1.2 eq.) Was added via syringe. The reaction was allowed to stir at room temperature overnight.

The clear red solution changed to cloudy after approximately two hours. TLC (10% MeOH in CH2C12) showed total loss of the initiator material. The reaction was transferred to a separatory funnel with EtOAc and washed with 40 mL of H20 followed by 40 mL of brine. The organic layer was collected and passed through MgSO4 in a vitrified glass funnel to dry it. The solvent was removed to give a purple solid (compound 39). A sample was analyzed by IS / MS. The IS / MS detected a MH * peak at 559, the total yield was 23 mg.

Conversion of Dithiocarbamate to the Trifluoromethyl Group 23 mg (0.041 mmol, 1 eq) of dithiocarbamate 39 was dissolved in anhydrous CH2C12 in a dry 25 mL round bottom flask equipped with stir bar, separators and nitrogen. The solution was cooled in an ice bath for approximately 15 min. Then 0.047 g of 1,3-dibromo-5,5-dimethylhydantoin (0.164 mmol, 4 eq.) Was added rapidly as a solid followed by 0.06 g of tetrabutylammonium dihydrogentrifluoride (0.205 mmol, 5 eq.) Via syringe. (The solution was reddish / purple to orange / brown).

The reaction was stirred at 0 ° C for 1.5 hours. It was then poured into a separatory funnel filled with 30 mL of H20. The CH2C12 layer was washed again with 25 ml of H20, collocated, and derivatized with MgSO4. The solvent was removed to give a dark brown / orange oil. The product was purified by the use of silica gel, starting with CH2C12 as the mobile phase and gradually adding methanol. Three separate points were obtained, the third sample was collected and the solvent was removed to provide 16.5 mg (75%) of the product 40.

Example 17 Acylation of the amine of Compound A 7 mg (0.0154 mmol) of Compound A was dissolved in 1 mL of dry CH2C12. It was collected at 0 ° C under N2, and stirred while adding 3.7 μL (0.046 mmol, 3 eq) of pyridine, followed by 2.06 μL (0.018 mmol, 1.2 eq.) Of trifluoroacetic anhydride (Aldrich). The reaction mixture was stirred at 0 ° C of 2:17 p.m. at 4:20 p.m. (TLC 1) and continued until 7:00 p.m. (TLC 2). There was no further progress of the reaction. Another bath of 3.7 μL of pyridine 2.6 μL of trifluoroacetic anhydride were added. After stirring for 4 h, TLC shows no significant progress of the reaction. The reaction mixture was allowed to warm to room temperature and was stirred for 3 h. Again, TLC does not show significant progress of the reaction. A third bath of 3.7 μL of pyridine and 2.0 mL of trifluoroacetic anhydride was added, the mixture was stirred for 2 h, TLC shows that the conversion is improved.

The reaction was stopped and the volatiles were evaporated under reduced pressure. The residue was dissolved in CHC13 and washed with saturated aqueous solution of NaHCO3 and water, twice and the CHC13 layer was evaporated. The residue was chromatographed on a column of silica gel eluted with 10% acetone-CHCl 3 (v / v) to obtain two pure components, 208.1 and 208.2. The remaining starter material in the column was eluted with CH3OH-CHC13 containing 10% Et3N, labeled 208.3. 208. 1 about 1 mg Rf = 0.67 208. 2 approximately 3 mg Rf = 0.45 208. 3 approximately 4 mg Rf = 0 'HR N in CDC13 208. 2 file: GZ .013 (Dept. 3): -N-CH ,, d 3.10 ppm. 208. 3 file: GZ .014 (Dept. 3): -N-CH3, d 2.40 ppm.

In CDC13, the methyl group signal shows a significantly small field that moves from 2.40 ppm to 3.10 ppm. Therefore the desired product is product 2 (compound 41).

The experiment was repeated as described below: A collection of recovered material 208.1, approximately 5 mg (0.011 mmol) was co-evaporated with toluene twice and dissolved in 2 mL of dry CH2C12 (with molecular sieve). 40 μL of pyridine (0.497 mmol, 45 eq) was added at 0 ° C, followed by 10 μL (0.071 mmol, 6.5 eq.) Of (CF3CO) 20). The mixture was stirred under N2 for 2 h, TLC 1 showed that the reaction was complete. The volatiles were evaporated in vacuo and the residue was passed through a small silicon gel column using 10% acetone-CHC13 as the eluent, yielding 5 mg of the product labeled 209-2.

Example 18 Preparation of bisindolyl maleimide of the formula: 0H Compound 24a, 140 mg (0.511 mmol) was evaporated with toluene twice and dissolved in 5.0 ml of fresh distilled anhydrous THF. It was cooled to 0 ° C (ice bath) and stirred under N2. 5.0 mL (5.0 mmol) of 1.0 M BH3-THF solution was added via a syringe. The resulting mixture was allowed to slowly warm to room temperature and was stirred under N2 for 15 h. It was cooled with an ice bath and then 10 ml of 10% NaOH was added, followed by the addition of 10 ml of 50% H202. The resulting cloudy white mixture was stirred at room temperature for 5 h. It was evaporated on a rotary evaporator to remove the THF and the residue was diluted with 50 ml of water. It was extracted with ethyl acetate (40 ml x 3). The ethyl acetate layer was washed with brine (50 mL) and dried with Na 2 SO 4. After evaporation the residue was separated on a column of silica gel (1 cm X 12.5 cm, flash chromatography) using toluene (20 ml), 30% -100% ethyl acetate in hexane (145 ml). The fourth component was identified to be the desired product 42a, 69 mg (46%).

Compound 49 was synthesized using the following steps and the same general procedure as discussed in detail in Example 14. 42a 43a 130mg (0. 5mmol) 197mg (98.8%) 45a 51mg (0.099mmol, 801) (0. 46a 46mg (0.094 iti ol, 95%) • 46mg to. DMF 47a HMDS 36mg (0.076-mmol 81%) CH3OH b. H + 48a 20mg (0.043 iraaol, 56%) 48a 49; Ejepplo 19 In Vitro Protein Kinase C Inhibition Test Reaction mixture: μl Ca + H "(reserve 9 .4 mM) 55 μl lipids (PS 5 μg / well, DG 0.6 μ / well) or HEPES μl Compound or DMSO (Test Compounds initial concentration at 5000 nM) 10 μl Substrate of Myelin Basal Protein (MBP) (MBP 3 mg / ml, lot # 451-026) μL ATP (300 μM ATP, 0.25 μCi / well AT32P, and 10 mM MgCl 2) μL Enzyme (PCC at 1:80 in HEPES, ß? R 1:30, in HEPES) HEPES buffer reserve is 100 mM, pH 7.5.

The Total Reaction Mixture was equal to 100 μl and incubated for 10 minutes at 30 ° C. The reaction was stopped with the addition of 100 μL 25% TCA. 25 μl of a 1 mg / ml BSA solution was added and 200 μl of the reaction mixture was transferred to a 96-well glass fiber filtration plate (Millipore Cat. # MAFCNOB50). The supernatant was filtered and washed three times with 10% TCA. The bottom of the filtration plate and the installed vehicle were removed. 100 μl of a Microscint-20 (Packard Cat. # 6013621) was added and the samples were loaded into a Packard counter.

Lipid preparation The lipids (Avanti Polar Lipids) were added to a borosilicate glass culture tube (25 x 150 mm). The lipids were dried under nitrogen until the chloroform was evaporated. The lipids were resuspended in the HEPES buffer, and sonicated for approximately 30 seconds, then forming a vortex to mix well. The lipids were kept on wet ice until the addition to the test.

Results The IC50 values were determined for each of the following compounds, using the above in vitro assay. Five concentrations of each compound were tested against alpha PCC (PCC or.) And PCC beta II (PCC ß "). The concentrations of the compounds tested were 5000 nM, 500 nM, 50 nM, 5 nM, 1 nM and zero (without compound, only DMSO). The sample without any added compound was used to determine 100% activity of the PCC enzyme in the test. The IC 50 values were estimated from the inhibition curves using these concentrations.

IC50 values in nanoHolar (nM) PKC to PKC ßa 57 5.0 Values IC5fl in nanoMolar (m ^) PKC to PKC ßa 79 5.5 IC5g values in nanoHolar (nM) PKC to PKC ßn 140 3.5 IC5g values in nanoHolar (nM) PKC to PKC ßa • 26 2.4 IC5g values in nanoHolar (nM) PKC to PKC ßa 2.6 2.2 IC5fl values in nanoHolar '(nM) PKC to KC ßn 2700 100 48a IC50 values in nanoHolar (nM) PKC to PKC ßa 130 13 49a IC50 values in nanoMolar (nM) KC to PKC ßp 1300 90 As an inhibitor of protein kinase C, the compounds set forth herein are useful in the treatment of conditions in which protein kinase C has demonstrated a role in the pathology. The conditions recognized in the art include: diabetes mellitus and its complications, ischemia, inflammation, disorders of the central nervous system, cardiovascular disease, Alzheimer's disease, dermatological disease and cancer.

Inhibitors of protein kinase C have been shown to block inflammatory responses such as oxidative neutrophil, low regulation of CD3 in T lymphocytes and phorbol-induced hand edema. Twoeny, B. et al. Biochem. Biophys. Res. Commun. 171: 1087-1092 (1990); Mulqueen, M.J. et al. Agents Actions 37: 85-89 (1992). Therefore, like the PCC inhibitors, the present compounds are useful in the treatment of inflammation.

The activity of protein kinase C plays a central role in the functioning of the central nervous system. Huang, K.P. Trends Neurosci. 12: 425-432 (1989). In addition, protein kinase C inhibitors have been shown to prevent damage seen in focal and central ischemic brain injury and cerebral edema. Hara, H. et al. J. Cereb. Blood Flow Metab. 10: 646-653 (1990); Shibata, S. et al. Brain Res. 594: 290-294 (1992). Recently, protein kinase C has been determined to be involved in Alzheimer's disease. Shimohama, S. et al., Neurology 43: 1407-1413 (1993). Therefore, the compounds of the present invention are useful in the treatment of Alzheimer's disease and ischemic brain injury.

The activity of protein kinase C has been associated with cell growth, tumor promotion and cancer. Rotenberg, S.A. and einstein, I.B. Biochem. Mol. Aspects Sel. Cancer l: 25-73 (1991). Ahmad et al., Molecular Pharmacology: 43 858-862 (1993). It is known that inhibitors of protein kinase C are effective in preventing the growth of tumors in animals. Meyer, T. et al. Int. J. Cancer 43: 851-856 (1989); Akinagaka, S. et al. Cancer Res. 51: 4888-4892 (1991). The compounds of the present invention also act as multidrug reverse agents (MDR) by making them effective compounds when administered in conjunction with other chemotherapeutic agents.

The activity of protein kinase C also plays an important role in cardiovascular disease. The activity of protein kinase C increased in vascularity has been shown to cause vasoconstriction and increased hypertension. A known inhibitor of protein kinase C prevents this increase. Bilder, G.E. et al. J. Pharmacal. Exp. Ther. 252: 526-530 (1990). Because inhibitors of protein kinase C demonstrate inhibition of oxidative neutrophil, inhibitors of protein kinase C are also useful in the treatment of cardiovascular ischemia and the improvement of cardiac function following ischemia. Muid, R.E. et al. FEBS Lett. 293: 169-172 (1990); Sonoki, H. et al. Kokyu-To Junkan 37: 669-674 (1989). The role of protein kinase C in platelet function has also been investigated and has shown high levels of protein kinase C that correlate with increased response to agonists. Bastyr III, E.J. and Lu, J. Diabetes 42: (Suppl 1) 97A (1993). PCC has been implicated in the biochemical pathway in the modulation of platelet activity factor of microvascular permeability. Kobayashi et al., Amer. Phys. Soc. H1214-H1220 (1994). Potent protein kinase C inhibitors have been shown to affect the induced aggregation of agonists in platelets. Toullec, D. et al. J. Biol. Chem. 266: 15771-15781 (1991). Inhibitors of protein kinase C also block cell proliferation of smooth muscle-induced agonist. Matsumoto, H. and Sasaki, Y. Biochem. Biophys. Res. Commun. 158: 105-109 (1989). Therefore, the present compounds are useful in the treatment of cardiovascular disease, atherosclerosis and restenosis.

The abnormal activity of protein kinase C has also been linked to dermatological disorders such as psoriasis. Horn, F. et al. J. Invest. Dermatol. 88: 220-222 (1987); Raynaud, F. and Evain-Brion, D. Br. J. Dermatol. 124: 542-546 (1991). Psoriasis is characterized by abnormal proliferation of keratinocytes. Known protein kinase C inhibitors have been shown to inhibit keratinocyte proliferation in a manner parallel to their potency as inhibitors of PCC. Hegemann, L. et al. Arch. Dermatol. Res. 286: 456-460 (1991); Bollag, W.B. et al. J. Invest. Dermatol. 100: 240-246 (1993). Therefore, the compounds as inhibitors of PCC are useful in the treatment of psoriasis. .

Protein kinase C has been linked to several different aspects of diabetes. Excessive activity of protein kinase C has been linked to insulin signaling defects and thus to the insulin resistance observed in type II diabetes. Karasik, A. et al. J. Biol. Chem. 265: 10226-10231 (1990); Chen, K.S. et al. Trans. Assoc. Am. Physicians 104: 206-212 (1991); Chin, J.E. et al. J. Biol. Chem. 268: 6338-6347 (1993). In addition, studies have shown a marked increase in the activity of protein kinase C in tissues known to be susceptible to diabetic complications when exposed to hyperglycemic conditions. Lee, T. S. et al. J. Clin. Invest. 83: 90-94 (1989); Lee, T.S. et al. Proc. Nati Acad. Sci. USA 86: 5141-5145 (1989); Craven, P.A. and DeRubertis, F.R. J. Clin. Invest. 83: 1667-1675 (1989); Wolf, B.A. et al. J. Clin. Invest. 87: 31-38 (1991); Tesfaramariam, B. et al. J. Clin. Invest. 87: 1643-1648 (1991).

The compounds of Formula I are preferably formulated before administration. Therefore, yet another aspect of the present invention is a pharmaceutical formulation comprising a compound of Formula I and one or more pharmaceutically acceptable carriers, diluents or excipients.

The present pharmaceutical formulations are prepared by known procedures using readily available and well-known ingredients. In making the compounds of the present invention, the active ingredient will usually be mixed with a vehicle, or diluted by a vehicle, or attached within a vehicle which could be in the form of a capsule, tablet, paper or other container. When the vehicle serves as a diluent, it could be a solid, semi-solid or liquid material which acts as a vehicle, excipient or medium for the active ingredient. Thus, the compositions may be in the form of tablets, pills, powders, lozenges, elixirs, suspensions, emulsions, solutions, syrups, aerosol (as a solid or in a liquid medium), soft and hard gelatin capsules, suppositories, solutions sterilized injectables and sterilized packaged powders.

Some examples of suitable vehicles, excipients and diluents include lactose, dextrose, sucrose, sorbitol, mannitol, starches, acacia gum, calcium phosphate, alginates, tragacanth, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water syrup , methyl cellulose, methyl and propylhydroxybenzoates, talc, magnesium stearate and mineral oil. The formulations may additionally include lubricating agents, moisturizing agents, emulsifying and suspending agents, preservatives, sweetening agents or flavoring agents. The compositions of the invention could be formulated to provide rapid, sustained or prolonged release of the active ingredient after administration to the patient. The compositions are preferably formulated in a dosage form, each dosage containing from about 1 to about 500 mg, more typically from about 5 to about 300 mg, of the active ingredient. However, it will be understood that the therapeutic dosage administered will be determined by the physician in the clarity of the relevant circumstances including the condition to be treated., the choice of the compound to be administered and the choice of the administration route. Therefore, the above dosage ranges are not intended to limit the scope of the invention in any way. The term "unit dosage form" refers to physically discrete units suitable as unitary dosages for human subjects and other mammals, each unit containing a predetermined quantity of the active material calculated to produce the desired therapeutic effect, in association with an appropriate pharmaceutical carrier. .

In addition to the above formulations, the compounds of the present invention could be administered topically. Topical formulations are ointments, creams, and gels. Ointments are generally prepared using either (1) an oil base, p. ex. , a base consisting of fixed oils or hydrocarbons, such as white petrolatum or mineral oil, or (2) an absorbent base, e.g. ex. , a base consisting of an anhydrous substance or substances that can absorb water, for example anhydrous lanolin. Usually, following the formation of the base, either oleoginous or absorbent, the active ingredient (compound) is added to an amount that provides the desired concentration.

The creams are oil / water emulsions. These consist of an oily phase (internal phase), typically 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 any soluble substance. in water, such as salts. The two phases are stabilized by the use of an emulsifying agent, for example, a surface active agent, such as sodium lauryl sulfate, hydrophilic colloids, such as acacia colloidal clays, gum V and the like. Due to the formation of the emulsion, the active ingredient (compound) is usually added to an amount to achieve the desired concentration.

The gels comprise a base selected from an oleoginous base, water, an emulsion-suspension base. A gelatinizing agent is added to the base which forms a matrix in the base, increasing its viscosity. Examples of gelling agents are hydroxypropyl cellulose, acrylic acid polymers, and the like. Usually, the active ingredient (compounds) is added to the formulation at the desired concentration at a point preceding the addition of the gelatinizing agent.

The amount of the compound that is incorporated in a topical formulation is not critical; the concentration should only be a sufficient range to allow adequate application of the formulation to the area of affected tissue in an amount that will release the desired amount of the compound.

The amount usually of a topical formulation to be applied to an affected tissue will depend on the size of tissue affected and the concentration of the compound in the formulation. In general, the formulation will be applied to the affected tissue in an amount that provides from about 1 to about 500 μg of the compound per cm 2 of an affected tissue. Preferably, the applied amount of the compound will depend on the range of. about 30 to about 300 μg / cm2, more preferably, 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 gelatin capsules are prepared using the following ingredients: The above ingredients are mixed and filled into hard gelatine capsules in amounts of 460 mg.

Formulation 2 A tablet is prepared using the following ingredients: The components are mixed and compressed to form tablets each weighing 665 mg.

Formulation 3 An aerosol solution is prepared containing the following components: The active compound is mixed with ethanol. The mixture is added to a portion of the propellant 22, cooled to -30 ° C and transferred to a filler device. The required amount is then fed to a stainless steel vessel and diluted with the remaining propellant. The valve units are then calibrated in the container.

Formulation 4 The tablets each containing 60 mg of the active ingredient are made as follows: The active ingredient, starch and cellulose are passed through a U.S. No. 45 mesh and mix thoroughly. The solution of polyvinyl pyrrolidone is mixed with the resulting powders which are then passed through a U.S. No. 14 mesh. The granules thus produced are dried at 50 ° C and passed through a U.S. No. 18. Sodium carboxymethyl starch, magnesium stearate and talc, previously passed through a U.S. No. 60 mesh, after adding to the granules which, after mixing, are compressed in a tabletting machine to produce the tablets each weighing 150 mg.

Formulation 5 The capsules each containing 80 mg of the drug are made as follows: The active ingredient, cellulose, starch and magnesium stearate are mixed, passed through a U.S. No. 45 mesh, and filled into hard gelatin capsules in amounts of 200 mg.

Formulation 6 Suppositories each containing 225 mg of the active ingredient could be made as follows: The active ingredient is passed through a U.S. No. 60 mesh and suspended in glycerides of saturated fatty acids previously melted using a minimum amount of heat. The mixture was then poured into a suppository mold of nominal 2 g capacity and allowed to cool.

Formulation 7 The suspensions each containing 50 mg of the drug per 5 mL of dose are made as follows: The medication is passed through a U.S. No. 45 mesh and mixed with microcrystalline sodium cellulose and syrup to form a smooth paste. The benzoic acid solution, the flavor and the color are diluted with some water and added, with stirring. Then enough water is added to produce the required volume.

Formulation 8 An intravenous formulation could be prepared as follows; The solution of the above ingredients is administered intravenously at a rate of 1 mL per minute to a subject in need of treatment.

It is noted that in relation to this date, the best method known by the applicant to carry out the aforementioned invention, is the conventional one for the manufacture of the objects to which it relates.

Having described the invention as above, the content of the following is claimed as property.

Claims (26)

1. A compound of the formula: characterized because: R 'is independently hydrogen, halo, hydroxy, alkyl C ,,, Cj.-C4 alkoxy, NR3R4 or -NHCO (C ^ -C- alkyl; V is -O-, -NH- or alkyl -NO.-C ,,; T is Ci-Q alkylene optionally substituted with halo or C3-C4 alkyl; W is C 2 alkylene optionally substituted with halo or Ci-G alkyl; J is • x-c- I s or when T and W are both methylene, J is selected from the group consisting of wherein n and m are independently 1 or 2; X is oxygen, sulfur or a bond between the carbon atoms joined by the X bridge; Y is halo, CX-C4 alkyl or hydrogen; Ri. Is hydrogen or Cj.-C4 alkyl; S is -CHO or the group wherein M is hydrogen, -CH20Rs, -CH2NR3R4 or -NR3R4; R2 is hydrogen or halo; Y Z is hydrogen or -0RS; wherein R 3 and R 4 are independently hydrogen, alkyl Q.-G, halo (C 1 -C 4 alkyl), C 1 -C 4 alkanoyl, halo (C 4 -C 4 alkanoyl) or R 3 and R 4 taken together with the N atom to which they are united they form a ring of 5 or 6 members; Y R5 and Rs are independently hydrogen, C-C-alkyl, halo (CL-C4 alkyl), CX-C4 alkanoyl, halo (C-C-alkanoyl or together form a divalent group selected from the group consisting of -CR7R8 - wherein R7 and R8 are independently hydrogen, Ci-C4 alkyl or halo (C ^ -C alkyl) or R7 and R8 taken together with the C atom to which they are attached form a 5 or 6 membered ring with the proviso that at least one of Y, S, T or W is halo or a substituted halo group, or T and W are both methylene.

2. The compound of claim 1, characterized in that at least one of Y, S, T or W is fluorine or a group substituted with fluorine.

3. A compound of the formula: characterized because: R. is Cj.-C4 alkyl or hydrogen; T is C2-C4 alkylene optionally substituted with halo or C ^ -C alkyl; W is ethylene optionally substituted with halo or alkyl X is oxygen, sulfur or a bond between the carbon atoms joined by the X bridge; Y. is halo, CX-C4 alkyl or hydrogen; Rj. is hydrogen or C 1 -C 4 alkyl; S is -CHO or the group wherein M is hydrogen, -CH2OR5, -CH2NR3R4 or -NR3R4; R2 is hydrogen or halo; Y Z is hydrogen or -0RS; wherein R 3 and R 4 are independently hydrogen, C 1 -C 4 alkyl, halo (C 4 -C 4 alkyl), CC 4 alkanoyl, halo (Ci-C- alkanoyl) or R 3 and R 4 taken together with the N atom to which they are united they form a ring of 5 or 6 members; Y Rs and Rs are independently hydrogen, CX-C4 alkyl, halo (C-C4 alkyl), alkanoyl GG, halo (C-alkanoyl or together form a divalent group selected from the group consisting of -CR7RB- wherein R7 and R8 are independently hydrogen, alkyl G.- or halo (G-C4 alkyl) or R7 and R8 taken together with the C atom to which they are attached form a 5- or 6-membered ring • with the proviso that at least one of Y, S, T or W is halo or a substituted halo group, or T and W are both methylene.

4. The compound of claim 3, characterized in that at least one of Y, S, T or W is fluorine or a group substituted with fluorine.

5. The compound of claim 3, characterized in that W is ethylene substituted with fluorine.

6. The compound of claim 3, characterized in that T is ethylene substituted with fluorine.

7. The compound of claim 3, characterized in that T is trimethylene substituted with fluorine.

8. The compound of claim 3, characterized in that ^ and Y are hydrogen and X is oxygen.

9. The compound of claim 5, characterized by RL and Y are hydrogen, X is oxygen and S is M I -CH I R2

10. The compound of claim 9, characterized in that T is ethylene.

11. A compound of the formula: characterized in that J is selected from the group consisting of wherein Rx is Cx-C4 alkyl or hydrogen; n and m are independently 1 or 2; Y R3 and R4 are independently hydrogen, alkyl G-G, halo (CX-C4 alkyl), CX alkanoyl = C4, halo (Cx-C4 alkanoyl) or taken together with the N atom to which they are attached form a 5- or 6-membered ring.

12. The compound of claim 9, characterized in that Rx is hydrogen.

13. The compound of claim 9, characterized in that the halo substituent of J is fluorine.

14. A pharmaceutical composition, characterized in that it comprises a compound of the formula: where; R1 is independently hydrogen, halo, hydroxy, CX-C4 alkyl, Cx-C4 alkoxy, NR3R4 or -NHCO (CX-C4 alkyl); R x is C 1 -C 4 alkyl or hydrogen; T is Cx-C4 alkylene optionally substituted with halo or Cx-C4 alkyl; is Cx-C2 alkylene optionally substituted with halo or CX-C4 alkyl; J. is Y I -x-c- I s or when T and W are both methylene, J is selected from the group consisting of wherein n and m are independently 1 or 2; X is oxygen, sulfur or a bond between the carbon atoms joined by the X bridge; Y is halo, CX-C4 alkyl or hydrogen; S is -CHO or the group L wherein M is hydrogen, -CH20R5, -CH2NR3R4 or -NR3R4; R2 is hydrogen or halo; Y Z is hydrogen or -0R6; wherein R3 and R4 are independently hydrogen, CX-C4 alkyl, halo (CX-C4 alkyl), CX-C4 alkanoyl, halo (Cx-C4 alkanoyl) or R3 and R4 taken together with the N atom to which they are attached form a 5 or 6 member ring; Y R5 and Rs are independently hydrogen, CX-C4 alkyl, halo (CX-C4 alkyl), CX-C4 alkanoyl, halo (CX-C4 alkanoyl) or together form a divalent group selected from the group consisting of -CR7R8- wherein R7 and R8 are independently hydrogen, CX-C4 alkyl or halo (CX-C4 alkyl) or R7 and R8 taken together with the C atom to which they are attached form a 5 or 6 membered ring with the proviso that at least one of Y, S, T or W is halo or a substituted halo group, or T and W are both methylene; and a pharmaceutically acceptable excipient, vehicle or diluent.

15. The pharmaceutical composition of claim 14, characterized in that J is And I -x-c-

16. The pharmaceutical composition of claim 15, characterized in that S is «/ -M 1, X is oxygen; Y Rx is hydrogen.

17. The pharmaceutical composition of claim 16, characterized in that at least one of Y, S, T or W is fluorine or a group substituted with fluorine.

18. The pharmaceutical composition of claim 14, # • characterized in that T and W are methylene and the halo substituent is fluorine.

19. A method of treating a mammal having a disease or condition associated with the abnormal activity of protein kinase C, characterized in that said method comprises administering to the mammal a pharmaceutically effective amount of the compound of the formula: where: R 'is independently hydrogen, halo, hydroxy, CX-C4 alkyl, Cx-C4 alkoxy, NR3R4 or -NHCO (CX-C4 alkyl); R x is C 1 -C 4 alkyl or hydrogen; T is Cx-C4 alkylene optionally substituted with halo or Cx-C4 alkyl; W is Cx-C2 alkylene optionally substituted with halo or Cx-C4 alkyl; J. is Y I • x-c- I s or when T and W are both methylene, J is selected from the group consisting of -t 143 wherein n and independently are 1 or 2; X is oxygen, sulfur or a bond between the carbon atoms joined by the X bridge; Y is halo, CX-C4 alkyl or hydrogen; S is -CHO or the group wherein M is hydrogen, -CH20R5, -CH2NR3R4 or -NR3R4; R2 is hydrogen or halo; Y Z is hydrogen or -0RS; wherein R3 and R4 are independently hydrogen, CX-C4 alkyl, halo (CX-C4 alkyl), CX-C4 alkanoyl, halo (Cx-C4 alkanoyl) or R3 and R4 taken together with the N atom to which they are attached form a 5 or 6 member ring; Y Rs and R6 are independently hydrogen, CX-C4 alkyl, halo (CX-C4 alkyl), CX-C4 alkanoyl, halo (Cx-C4 alkanoyl) or together form a divalent group selected from the group consisting of -CR7Ra- wherein R7 and R8 are independently hydrogen, CX-C4 alkyl or halo (CX-C4 alkyl) or R7 and R8 taken together with the C atom to which they are attached form a 5 or 6 membered ring with the proviso that at least one of Y, S, T or W is halo or a substituted halo group, or T and W are both methylene; and a pharmaceutically acceptable excipient, vehicle or diluent.

20. The method of claim 19, characterized in that J is And I • x-c-

21. The method of claim 20, characterized in that S is / -M fc X is oxygen; Y Rx is hydrogen.

22. The method of claim 20, characterized in that at least one of Y, S, T or W is fluorine or a group substituted with fluorine.

23. The method of claim 19, characterized in that T and W are methylene and the halo substituent is fluorine.

24. The method of claim 19, characterized in that W is ethylene substituted with fluorine.

25. The method of claim 19, characterized in that T is ethylene substituted with fluorine.

26. The method of claim 19, characterized in that T is trimethylene substituted with fluorine.