MXPA99002255A - Inhibition of 26s and 20s proteasome by indanones - Google Patents

Abstract

This invention is a method for inhibiting cell proliferation using indanones.

Description

INHIBITION OF PROTEASOMA 26S AND 20S BY INDANONAS BACKGROUND OF THE INVENTION 1. Field of the Invention This invention is a method for inhibiting cell proliferation using a class of indanone compositions never before considered for that purpose. As inhibitors of cell proliferation, the compositions are useful in the treatment of cancer, cardiovascular diseases, for example, restenosis, rejection of host grafts, gout and other proliferative diseases as well as being potential therapeutics for autoimmune diseases, such as rheumatic arthritis, lupus, type I diabetes, multiple sclerosis and similar disorders and diseases. 2. Description of the Technique Proteinase or multicatalitic proteasome is a highly conserved cell structure that is responsible for the ATP-dependent proteolysis of most cellular proteins. The 20S proteasome (700-kDa) contains at least five proteolytic activities that have a new type of mechanism that involves a threonine residue in the active site (Coux O., Tanaka K. and Goldberg, A. 1996 Ann. Rev. Biochem 65: 801 -47).

The 20S proteasome has been crystallized from the archaeobacterium Thermoplasma acidophilum (Lowe, J., Stock, D., Jap, B., Swickl, P., Bauminster, W., and Huber, R., 1995 Science 268: 533 539). The arcaebacterial 20S proteasome contains fourteen copies of two distinct types of subunits «and ß, which form a cylindrical structure of four stacked chains. The upper and lower chains contain seven subunits 8 each, while the inner chains contain seven β subunits. A pore extends through the middle of the structure containing the active poteolytic sites and the proteins destined for degradation passing through this channel. The eukaryotic 20S proteasome is more complex than that of the archaebacterium because the number of different subunits has increased during evolution, however, the subunits can still be classified according to the QC nomenclature and ß of arcaebacteria according to their homology. Thus the quaternary structure of the eukaryotic complex is similar to that of the archaebacteria being composed of two chains oc and two chains ß. However, unlike the arcaebacterial proteasome that mainly exhibits proteolytic activity similar to chymotrypsin (Dahlmann, B., Kopp, F., Kuehn, L., Niedel, B., Pfeifer, G, 1989 FEBS 251: 125-131, Seemuller, E., Lupas, A., Zuhl, F., Swickl, P. and Baumeister, W. 1995 FEBS Lett 359: 173, and Lowe, J., Stock, D., Jap, B., Swickl, P., Bauminster, W. and Huber, R., 1995 Science 268: 533-539). The eukaryotic proteasome contains at least five identifiable protease activities. These are called peptidylglutamyl-peptide hydrolysates similar to chymotrypsin and trypsin-like. Two other activities have been described, one exhibiting a preference for the division of peptide bonds on the carboxyl side of amino acids with branched chain and the other towards the bonds between small neutral amino acids. (Orlowski, M. 1990 Biochemistry 29: 10289-10297).

Although the 20 S proteasome contains the proteolytic core, it can not degrade proteins in vivo unless it is complexed with a 19S cap at either end of its structure, which itself contains multiple ATPase activities. This larger structure is known as the 26S proteasome and rapidly degrades the proteins that have been chosen for degradation by the addition of multiple molecules of the 8.5-kDa polypeptide, ubiquitin.

The first step towards the ubiquitination of a protein proceeds through the activation of a ubiquitin molecule in its glycine residue at the carboxyl terminus by the addition of ATP which creates a high energy thioester intermediate. This step is catalyzed by the ubiquitin activation enzyme, El. The ubiquitin is then transferred to the active cysteine residue of an enzyme that conjugates ubiquitin, E2. The E2 enzymes couple ubiquitin to the E-amino groups of lysine residues in the substrate protein that is destined to be degraded. This process, in some cases also requires a ubiquitin ligase, E3. Repeated conjugation of ubiquitin to lysine residues of previously bound ubiquitin halves leads to the formation of multi-ubiquitin chains and creates a ubiquitin region around the substrate protein. The multibiquitinated proteins are recognized by the 26 S proteasome and are degraded and the multiubiquitin chains are released from the complex and the ubiquitin is recycled.

What causes a protein to become uniquitinated and thus degrade is still under investigation. Clearly this must be a highly regulated series of events since the critical coordination of specific protein degradation is crucial for many cell cycle functions. Several signals have been proposed that largely focus on the internal structural sequences within the substrate itself. One such proposal is the "extreme rule N" in which the amino terminal residue of a protein determines its half-life. Other proteins such as cyclins contain a short sequence of highly conserved amino acids called the "destruction box" that are apparently necessary for degradation. In addition, the "PEST" sequences, which consist of regions rich in proline, aspartate, glutamate, serine and threonine, appear to act as degradation signals. It is thought that said internal sequences act as recognition elements between the protein substrate and its specific ubiquitination machinery.

Two types of inhibitors that inhibit the proteolytic activity of the proteasome have been described. It has been reported that certain peptide aldehydes inhibit chymotrypsin-like activity related to the proteasome (Vinitsky, A., Michaud, C, Powers, J., and Orlowski, M. 1992 Biochemistry 31: 9421-9428; Tsubuki, S., Hiroshi, K., Saito, Y., Miyashita, N., Inomata, M., and Kawashima, S., 1993 Biochem, Biophys, Res. Commun. 196: -1195-1201; Rock, KL, Gramm. C, Rothstein, L., Clak, K., Stein, R., Dick, L., Hwang, D., and Goldberg, AL 1994 Cell 78: 761-771). These are N-acetyl-L-leucinyl-L-leucinal-L-norleucinal (ALLN) and a closely related compound, N-acetyl - L - leucinil - leucinil - metional (LLM) with a K¡'s of 0.14 μM. The most potent inhibitor of this type is a structurally related compound, N-carbobenzoxyl-L-leucinil - L - norvalinal (MG 115) exhibiting a KI of 0.021 μM. Although these peptide aldehydes are more effective against the proteolytic activity similar to chymotrypsin of proteasomes, careful studies have shown that they are not specific protease inhibitors. The most recent reports have described series of potent dipeptide inhibitors that have IC 50 values in the range of 10-100 nM in vitro (Iqbal, M., Chaterjee, S., Kauer, JC, Das, M., Messina, P., Freed, B., Biazzo, W., and Siman, R., 1995 J. Med. Chem. 38: 2276-2277) and a series of potent in vivo inhibitors in a similar manner from di-peptides derived from oc-ketacarbonyl and boronic ester (Iqbal, M., Chaterjee, S., Kauer, JC, Mallamo, JP, Messina, PA, Reibolt, A., and Siman, R., 1996 Bioorg, Med Chem Lett 6: 287-290 ).

Another report describes a class of compounds exhibiting specificity in the inhibition of proteasome activity (Fenteany, G, Standaert, R.F., Lane, W.S., Choi, S., Corey, E.J., and Schreiber, S.L. 1995 Science 268: 726-731). Lactacystin is a metabolite Strepmyces that specifically inhibits the proteolytic activity of the proteasome complex. This molecule was originally discovered for its ability to induce neurite overgrowth in a neuroblastoma cell line (Ormura et al., 1991 J. Antibiot 44: 113) it was later shown to inhibit the proliferation of several cell types (Fenteany, et al. al., 1994 Proc. Nat'l. Acad. Sci. USA 91: 3358). Through the use of radiolabeled lactacistin, link studies by (Fenteany, et al., 1995 Science 268: 726-731) have identified the binding site and the mechanism of action. These studies have shown that lactacystin binds irreversibly to a threonine residue located at the amino terminus of the β subunit of proteasomes. A series of analogues based on the lactacystin structure were also investigated (Fenteany, et al., 1995 Science 268: 726-731). These studies indicated that the β-lactone structure was essential for its inhibitory activity.

It is now well established that the proteasome is a major extralisosomal proteolytic system that is involved in degrading pathways that result in numerous and diverse cellular functions such as cell division, antigen processing and the degradation of short-lived regulatory proteins such as of transcription, oncogene products and cyclins (reviewed in Ciechanover, A., 1994 Cell 79:13 - 21). The primary function of the proteasome is to catalyze protein proteolysis in small peptides. However, it has also been shown that the ubiquitin-proteasome pathway can catalyze the regulated proteolytic processing of a large inactive precursor to an active protein. The best documented case of the above involves the activation of the NF-KB transcription factor (Palombella, V.J., Rando, O.J., Goldberg, A.L., and Maniatis, T., 1994 Cell 78: 773-785). The active form of NF-KB is a heterodimer consisting of a p65 subunit and a p50 subunit. The latter is present in the sitosol of the cell in an inactive precursor of, primarily pl05, the 105-kDa polypeptide precursor of p50. The proteolytic processing of pl05 to generate p50 occurs via the ubiquitin-proteasome pathway. In addition, p50 and p65 processed remain in the cytosol as an inactive complex with the inhibitor protein I? B. Inflammatory signals activate NF-KB by initiating the signaling pathway for complete degradation of I? B, and also stimulate p 105 to p50 processing. Thus, two proteolytic events, both governed by the ubiquitin-proteasome pathway, are required for the activation induced by NF-KB signals. What causes the termination of pl05 proteolysis after generation of p50 is not known but it has been proposed that the conformation of p50 can make it resistant to other processing and cause it to be disassociated from the 26S complex.

The fact that the proteasome plays a critical event in the activation of NF-KB could be exploited clinically by the use of inhibitors directed towards the proteolysis of proteasomes. In certain diseases the normal function of active NF-KB can be harmful to human health as seen in the inflammatory responses that follow bacterial, fungal or viral infection. Thus, inhibitors of F-KB activation, due to their ability to prevent cytokine secretion, may have potential utility in the treatment of ARDS (acute respiratory distress syndrome) and SE) A. Since the activation of NF - KB is also essential for angiogenesis, proteasome inhibitors may have utility in the treatment of those diseases related to abnormal neovascularization. p53 was first described as an oncoprotein but has since been shown to be involved in many cellular processes (reviewed by Ko, L.J.

Proves, C. 1996 Genes Dev. 10, 1054-1072). It has been shown that P53 induces apoptosis in several haematopoietic cell lines (Oren, M., 1994 Semin. Cancer Biol. 5, 221-227) through the action of many different stimuli including DNA damage, viral infection and elimination of growth factors. However, it is important to note that apoptosis can be induced in a p53-dependent manner for example by the action of glucocoricoids. The induction of p53 leads to the arrest of cell growth in the Gl phase of the cell cycle as well as cell death by apoptosis. These two functions allow p53 to control DNA damage thereby reducing the spread of DNA mutations when cells divide. p53 arrests the cells in Gl by inducing the cyclin-dependent kinase inhibitor, p21, which in turn causes an accumulation of the hypophosphorylated form of the retinoblastoma gene product. It is thought that p53 acts as a checkpoint in the cell after DNA damage, first causing an arrest in cell division and apoptosis. It is known that the degradation of p53 is via the ubiquitin-proteasome pathway and interrupting the degradation of p53 is a possible way to induce apoptosis. Another potential utility of proteasome inhibitors may be in the treatment of diseases resulting from the abnormal proliferation of cells.

It is well documented that the ubiquitin-proteasome pathway is critical for the regulated destruction of cyclins that govern the exit of mitosis and allows cells to progress to the next phase of the cell cycle. Thus the inhibition of the degradation of cyclins by the use of proteasome inhibitors causes an arrest of growth. Therefore, another potential utility of proteasome inhibitors is their use in the treatment of diseases resulting from accelerated cell division. These include cancer, cardiovascular diseases such as myocarditis, restenosis after angioplasty, kidney diseases such as lupus, polycystic kidney disease, infections with fungi, dermatological diseases such as psoriasis, abnormal wound healing, keloids, immunological diseases such as autoimmunity, asthma and allergy, acute and delayed hypersensitivity, graft-versus-host disease, transplant rejection and neuroimmunological diseases such as multiple sclerosis and disseminated encephalomyelitis.

SUMMARY OF THE INVENTION It is an objective of this invention to provide a method for inhibiting mammalian cell proliferation that utilizes a therapeutically effective amount of a composition hitherto unknown for its proliferative inhibition properties.

It is an object of this invention to provide a method for the effective treatment of diseases resulting from accelerated cell division.

Another object of this invention is to provide a method for the treatment of proliferative diseases that operates by inhibiting the degradation of proteasome inhibitors.

Another objective of this invention is to use a therapeutically effective amount of the composition to inhibit cell proliferative disorders in humans.

In one example, this invention is a method for inhibiting mammalian cell proliferation comprising administering to the mammal a therapeutically effective amount of a compound having the formula: In the compound, Ri - »are each individually selected from the group including hydrogen, halogen, hydroxyl, thiol, lower alkyl, substituted lower alkyl, alkenyl, alkynyl, alkyl alkenyl, alkyl alkynyl, alkoxy, alkylthio, acyl, aryloxy, amino, amido, carboxyl, aryl, substituted aryl, heterocycle, heteroaryl, substituted heterocycle, heteroalkyl, cycloalkyl, substituted cycloalkyl, alkylcycloalkyl, alkylcycloheteroalkyl, nitro or cyano.

R5-R9 are each independently selected from the group of compounds including hydrogen, halogen, hydroxyl, thiol, oxo, lower alkyl, substituted lower alkyl, alkenyl, alkynyl, alkyl alkenyl, alkyl alkynyl, alkoxy, alkylthio, acyl, aryloxy, amino, amido, carboxyl, aryl, substituted aryl, heterocycle, heteroaryl, substituted heterocycle, heteroalkyl, cycloalkyl, substituted cycloalkyl, alkylcycloalkyl, alkylcycloheteroalkyl, nitro or cyano; X is selected from the group of compounds including hydrogen, -Di, -D2, -E, -Di-E, -D2-E, -Di-D2 or a compound having the formula: / 1 wherein Di and D2 are each independently chosen from the group of compounds that include a compound having the formula or hydrogen, halogen, thiol, lower alkyl, substituted minor alkyl, alkenyl, alkynyl, alkyl alkenyl, alkyl alkynyl, alkoxy, alkylthio, acyl, aryloxy, amino, amido, carboxyl, aryl, substituted aryl, heterocycle, heteroaryl, substituted heterocycle, heteroalkyl , cycloalkyl, substituted cycloalkyl, alkylcycloalkyl, alkylcycloheteroalkyl; wherein E is chosen from the group of compounds that includes: or hydrogen, halogen, thiol, lower alkyl, substituted minor alkyl, alkenyl, alkynyl, alkyl alkenyl, alkyl alkynyl, alkoxy, alkylthio, acyl, aryloxy, amino, amido, carboxyl, aryl, substituted aryl, heterocycle, heteroaryl, substituted heterocycle, heteroalkyl , cycloalkyl, substituted cycloalkyl, alkylcycloalkyl, alkylcycloheteroalkyl; When Di, D2 and / or E are chosen from the compounds that include Rio-R substitutes? , Ji and J2, then Rio-RH are each independently selected from the group of compounds including hydrogen, halogen, hydroxyl, thiol, oxo, lower alkyl, substituted lower alkyl, alkenyl, alkynyl, alkyl alkenyl, alkyl alkynyl, alkoxy, alkylthio , acyl, aryloxy, amino, amido, carboxyl, aryl, substituted aryl, heterocycle, heteroaryl, substituted heterocycle, heteroalkyl, cycloalkyl, substituted cycloalkyl, alkylcycloalkyl, alkylcycloheteroalkyl, nitro or cyano and Ji and J2 are each independently selected from the group of compounds including N-R15, CR? 6 R17, O, S - (O) or -2, P - (O) 0-3, where R15 - R7 can each be chosen independently from the group of compounds including hydrogen, halogen, hydroxyl, thiol, oxo, lower alkyl, substituted lower alkyl, alkenyl, alkynyl, alkyl alkenyl, alkyl alkynyl, alkoxy, alkylthio, acyl, aryloxy, amino, amido, carboxyl, aryl, substituted aryl, heterocycle , heteroaryl, heterocycle substituted, heteroalkyl, cycloalkyl, substituted cycloalkyl, alkylcycloalkyl, alkylcycloheteroalkyl or cyano.

The compositions are useful, when administered in therapeutic amounts, to treat mammals, and preferably to treat humans suffering from cell proliferative disorders, infectious diseases and immunological diseases.

BRIEF DESCRIPTION OF THE DUCTS In the figures, Figure 1 is a Western blot immunoreactivity assay using an anti-I? B antibody of RAW cell extract that had been treated with compounds 173 and 187 which are described in Tables 1 and 2; Figure 2 is a Western blot immunoreactivity assay towards an anti P50 antibody of RAW cell extracts that had been treated with the compound 187 which is described in Tables 1 and 2 before being exposed to its LPS; Y Figure 3 is a gel mobility change assay using nuclear extract prepared from RAW cells that had been pretreated with compound 187 which is described in Tables 1 and 2 before exposure to LPS.

DESCRIPTION OF THE CURRENT EXAMPLE This invention is a method for inhibiting disorders of cell proliferation, infectious diseases and immunological diseases in mammals and especially in humans using compositions having the following general formula: In the composition, Ri-R 'are each individually selected from the group including hydrogen, halogen, hydroxyl, thiol, lower alkyl, substituted lower alkyl, alkenyl, alkynyl, alkyl alkenyl, alkyl alkynyl, alkoxy, alkylthio, acyl, aryloxy, amino , amido, carboxyl, aryl, substituted aryl, heterocycle, heteroaryl, substituted heterocycle, heteroalkyl, cycloalkyl, substituted cycloalkyl, alkylcycloalkyl, alkylcycloheteroalkyl, nitro or cyano.

In the composition, R5-R are each independently chosen from the group of compounds including hydrogen, halogen, hydroxyl, thiol, oxo, lower alkyl, substituted lower alkyl, alkenyl, alkynyl, alkyl alkenyl, alkyl alkynyl, alkoxy, alkylthio, acyl, aryloxy, amino, amido, carboxyl, aryl, substituted aryl, heterocycle, heteroaryl, substituted heterocycle, heteroalkyl, cycloalkyl, substituted cycloalkyl, alkylcycloalkyl, alkylcycloheteroalkyl, nitro or cyano; X is selected from the group of compounds including hydrogen, - Di, -D2, -E, Di-D2, -Di-E, -D2-E, - or a compound having the formula: wherein Di and D2 are each independently chosen from the group of compounds that include a compound having the formula or hydrogen, halogen, thiol, lower alkyl, substituted minor alkyl, alkenyl, alkynyl, alkyl alkenyl, alkyl alkynyl, alkoxy, alkylthio, acyl, aryloxy, amino, amido, carboxyl, aryl, substituted aryl, heterocycle, heteroaryl, substituted heterocycle, heteroalkyl , cycloalkyl, substituted cycloalkyl, alkylcycloalkyl, alkylcycloheteroalkyl; wherein E is chosen from the group of compounds that includes: or hydrogen, halogen, thiol, lower alkyl, substituted minor alkyl, alkenyl, alkynyl, alkyl alkenyl, alkyl alkynyl, alkoxy, alkylthio, acyl, aryloxy, amino, amido, carboxyl, aryl, substituted aryl, heterocycle, heteroaryl, substituted heterocycle, heteroalkyl , cycloalkyl, substituted cycloalkyl, alkylcycloalkyl, alkylcycloheteroalkyl; In the compounds identified above, R5-R9 are each independently selected from the group of compounds including hydrogen, halogen, hydroxyl, thiol, oxo, lower alkyl, substituted lower alkyl, alkenyl, alkynyl, alkyl alkenyl, alkyl alkynyl, alkoxy, alkylthio, acyl, aryloxy, amino, amido, carboxyl, aryl, substituted aryl, heterocycle, heteroaryl, substituted heterocycle, heteroalkyl, cycloalkyl, substituted cycloalkyl, alkylcycloalkyl, alkylcycloheteroalkyl, nitro or cyano; When Di, D2 and / or E are chosen from the compounds including Rio-R? 4 substitutes, Ji and J2, then Rio - R? are each independently selected from the group of compounds including hydrogen, halogen, hydroxyl, thiol, oxo, lower alkyl, substituted lower alkyl, alkenyl, alkynyl, alkyl alkenyl, alkyl alkynyl, alkoxy, alkylthio, acyl, aryloxy, amino, amido , carboxyl, aryl, substituted aryl, heterocycle, heteroaryl, substituted heterocycle, heteroalkyl, cycloalkyl, substituted cycloalkyl, alkylcycloalkyl, alkylcycloheteroalkyl, nitro or cyano and Ji and 2 are each independently selected from the group of compounds including N-R15, CR? 6 Rp, O, S - (O) or -2, P - (O) or .3, wherein R15 - Rp can each be chosen independently from the group of compounds including hydrogen, halogen, hydroxyl, thiol, oxo, lower alkyl, substituted minor alkyl, alkenyl, alkynyl, alkylalkenyl, alkyl alkynyl, alkoxy, alkylthio, acyl, aryloxy, amino, amido, carboxyl, aryl, substituted aryl, heterocycle, heteroaryl, substituted heterocycle, heteroalkyl, cycloalkenyl, chloralkyl, substituted cycloalkyl, alkylcycloalkyl, alkylcycloheteroalkyl or cyano.

The following terms are used to describe various constituents of the chemical composition useful in the method of this invention. The terms are defined as follows: The term "halogen" refers to fluorine, bromine, chlorine and iodine atoms.

The term "hydroxyl" refers to the group-OH.

The term "oxo" refers to the group = O.

The term "thiol" or "mercapto" refers to the group -SH, and -S (O) o-2.

The term "lower alkyl" refers to a cyclic, branched or straight chain, alkyl group of one to ten carbon atoms. This term is further exemplified by such groups as methyl, ethyl, n-propyl, I-propyl, n-butyl, t-butyl, I-butyl (or 2-methylpropyl), cyclopropylmethyl, I-amyl, n-amyl, hexyl and similar.

The term "substituted lower alkyl" refers to lower alkyl as just described including one or more groups such as hydroxyl, thiol, alkylthiol, halogen, alkoxy, amino, amido, carboxyl, cycloalkyl, substituted cycloalkyl, heterocycle, cycloheteroalkyl, substituted cycloheteroalkyl. , acyl, carboxyl, aryl, substituted aryl, aryloxy, hetaryl, substituted hetaryl, aralkyl, heteroaralkyl, alkenyl alkyl, alkynyl, alkyl, cycloalkyl, cycloheteroalkyl, cyano. These groups can be attached to any carbon atom of the minor alkyl moiety.

The term "alkenyl" refers to a group -CR '= CR "R" wherein R', R ", R" 'are each selected from hydrogen, halogen, lower alkyl, substituted lower alkyl, acyl, aryl, aryl substituted, heteroaryl, substituted heteroaryl or the like as defined.

The term "alkynyl" refers to a group-C = C-R '; wherein R 'is selected from hydrogen, halogen, lower alkyl, substituted lower alkyl, acyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl or the like as defined.

The term "alkyl alkenyl" refers to a group -R-CR '= CR' '' R '"', wherein R is minor alkyl or substituted minor alkyl, R ', R'", R "" are selected each one independently of hydrogen, halogen, lower alkyl, substituted lower alkyl, acyl, aryl, substituted aryl, hetaryl or substituted hetaryl as defined below.

The term "alkynyl alkyl" refers to a group-RC = CR 'wherein R is minor alkyl or substituted minor alkyl, R' is hydrogen, minor alkyl, substituted minor alkyl, acyl, aryl, substituted aryl, hetaryl or substituted hetaryl as defined later.

The term "alkoxy" refers to the group -OR, wherein R is minor alkyl, substituted minor alkyl, acyl, aryl, substituted aryl, aralkyl, substituted aralkyl, heteroalkyl, heteroarylalkyl, cycloalkyl, substituted cycloalkyl, cycloheteroalkyl or substituted cycloheteroalkyl as define ahead.

The term "alkylotium" denotes the group - SR, - S (O) "=? .2-R, wherein R is minor alkyl, substituted minor alkyl, aryl, substituted aryl, aralkyl or substituted aralkyl as defined below.

The term "acyl" refers to the groups -C (O) R, wherein R is hydrogen, lower alkyl, substituted minor alkyl, aryl, aryl substituted and the like as defined below.

The term "aryloxy" refers to the groups -OAr, where Ar is an aryl, substituted aryl, heteroaryl or substituted heteroaryl group as defined below.

The term "amino" refers to the group NRR ', wherein R and R' may independently be hydrogen, lower alkyl, substituted lower alkyl, aryl, substituted aryl, hetaryl, cycloalkyl, or substituted hetaryl as defined below or acyl .

The term "amido" refers to the group-C (O) NRR ', wherein R and R' may independently be hydrogen, lower alkyl, substituted minor alkyl, aryl, substituted aryl, hetaryl, substituted hetaryl as defined below .

The term "carboxyl" refers to the group-C (O) OR, wherein R can independently be hydrogen, lower alkyl, substituted minor alkyl, aryl, substituted aryl, hetaryl, substituted hetaryl and the like as defined.

The term "aryl" or "Ar" refers to an aromatic carbocyclic group having at least one aromatic chain (e.g., phenyl or biphenyl) or multiple condensed chains in which at least one chain is aromatic (e.g., 1, 2,3"4-tetrahydronaphthyl, naphthyl, anthryl or phenanthryl).

The term "substituted aryl" refers to aryl optionally substituted with one or more functional groups, for example, halogen, lower alkyl, lower alkoxy, lower alkylthio, trifluoromethyl, amino, amido, carboxyl, hydroxyl, aryl, aryloxy, heterocycle, heteroaryl. , substituted hetaryl, nitro, cyano, alkylthio, thiol, sulfamido and the like.

The term "heterocycle" refers to a saturated, unsaturated or aromatic carbocyclic group having a single chain (eg, morpholino, pyridyl or furyl) or multiple condensed chains (e.g., napthyridyl, quinoxalyl, quinolinyl, indolizinyl or benzo [ b] thienyl) and having at least one hetero atom, such as N, O or S, within the chain, which may optionally be unsubstituted or substituted with, for example, halogen, lower alkyl, lower alkoxy, lower alkylthio, trifluoromethyl, amino, amido, carboxyl, hydroxyl, aryl, aryloxy, heterocycle, heteroaryl, substituted hetaryl, nitro, cyano, alkylthio, thiol, sulfamido and the like.

The terms "heteroaryl" or "hetar" refer to a heterocycle in which at least one heterocyclic chain is aromatic.

The term "substituted heteroaryl" refers to an optionally mono or polysubstituted heterocycle with one or more functional groups, eg, halogen, lower alkyl, lower alkoxy, lower alkylthio, trifluoromethyl, amino, amido, carboxyl, hydroxyl, aryl, aryloxy, heterocycle, hetaryl, substituted hetaryl, nitro, cyano, alkylthio, thiol, sulfamido and the like.

The term "aralkyl" refers to the group -R-Ar wherein Ar is an aryl group and R is lower alkyl group or substituted minor alkyl. The aryl groups may optionally be substituted or unsubstituted with, for example, halogen, lower alkyl, alkoxy, alkyl thio, trifluoromethyl, amino, amido, carboxyl, hydroxyl, aryl, aryloxy, heterocycle, heteroaryl, substituted hetaryl, nitro, cyano, alkylthio, thiol, sulfamido and the like.

The term "heteroalkyl" refers to the group -R-Het wherein Het is a heterocycle group and R is a lower alkyl group. Heteroalkyl groups can be unsubstituted or substituted with, for example, halogen, lower alkyl, lower alkoxy, lower alkylthio, trifluoromethyl, amino, amido, carboxyl, hydroxyl, aryl, aryloxy, heterocycle, heteroaryl, substituted hetaryl, nitro, cyano , alkylthio, thiol, sulfamido and the like.

The term "heteroarylalkyl" refers to the group -R-HetAr wherein HetAr is a heteroaryl group and R lower alkyl or substituted lower alkyl. The heteroaralkyl groups may be optionally unsubstituted or substituted with, for example, halogen, lower alkyl, substituted minor alkyl, alkoxy, alkylthio, aryl, aryloxy, heterocycle, heteroaryl, substituted hetaryl, nitro, cyano, alkylthio, thiol, sulfamido and the like .

The term "cycloalkyl" refers to a divalent polycyclic or cyclic alkyl group containing from 3 to 15 carbons. For polycyclic groups, these may be multiple condensed chains in which at least one of the distant chains may be aromatic (e.g., indanyl, tetrahydronaphthalene, etc.).

The term "substituted cycloalkyl" refers to a cycloalkyl group comprising one or more substitutes with, for example, halogen, lower alkyl, substituted minor alkyl, alkoxy, alkylthio, aryl, aryloxy, heterocycle, heteroaryl, substituted hetaryl, nitro, cyano , alkylthio, thiol, sulfamido and the like.

The term "cycloheteroalkyl" refers to a cycloalkyl group in which one or more of the chain carbon atoms is replaced with a heteroatom (eg, N, O, S or P).

The term "substituted cycloheteroalkyl" refers to a cycloheteroalkyl group as defined herein that contains one or more substitutes, such as halogen, lower alkyl, lower alkoxy, lower alkylthio, trifluoromethyl, amino, amido, carboxyl, hydroxyl, aryl, aryloxy, heterocycle, hetaryl, substituted hetaryl, nitro, cyano, alkylthio, thiol, sulfamido and the like.

The term "cycloalkyl alkyl" refers to the group -R-cycloalkyl wherein cycloalkyl is a cycloalkyl group and R is a lower alkyl or substituted lower alkyl. Cycloalkyl groups can be optionally substituted or unsubstituted with, for example, halogen, lower alkyl, lower alkoxy, lower alkylthio, trifluoromethyl, amino, amido, carboxyl, hydroxyl, aryl, aryloxy, heterocycle, heteroaryl, substituted hetaryl, nitro, cyano , alkylthio, thiol, sulfamido and the like.

The term "amino acid" refers to the D - or L - isomer of natural occurrence and the synthetic alpha amino acids, preferably the naturally occurring amino acids alanine, arginine, asparagine, aspartic acid, cysteine, glutamine, glutamic acid, glycine, histadine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine or valine.

Typically Di, D2 and E, if present in the composition, can be amino acids. Usually lipophilic amino acids are preferred. In general, amino acid abbreviations follow the IUPAC-IUB United Commission in Biochemical Nomenclature as described in Eur. J. Biochem, 158, 9 (1984).

It is preferred that R3 is methoxy, -Di is leucine and -D2 is leucine and E is NR'R ".

It is more preferred that -Di is 1 -leucine and -D2 is d-leucine and that E is chosen from the group consisting of benzylamine, 1-indanylamine, N, N'-dibenzylamine, 2,6-difluorobenzylamine, 4- methoxybenzylamine, piperonyl amine, NH2 and glycineamide.

In a preferred composition R3 is methoxy, Di is leucine, D2 is leucine and E is benzylamine. In another preferred composition, R3 is methoxy, Di is leucine, D2 is leucine and E is 1-indanylamine. In still another preferred composition R3 is methoxy, Di is leucine, D2 is leucine and E is N, N-debenzylamine. In another preferred composition, R3 is methoxy, Di is leucine, D is leucine and E is 2,6-difluorobenzylamine.

In these preferred compositions, it is further preferred that Di is L-leucine and D2 is D-leucine. The known compounds that may be useful in the therapeutic method of this invention are presented in Table 1 below.

TABLE 1 It is within the knowledge of one skilled in the art that the stereoisomers of the compositions described herein as well as the isomers and stereoisomers of components comprising the compositions identified herein all fall within the scope of the compositions that are useful in the therapeutic method of this invention. invention.

If the compound useful in the method of this invention contains a basic group, an acid addition salt can be prepared. The acid addition salts of the compounds are prepared in a standard manner in an appropriate solvent from the main compound and in excess of acid, such as hydrochloric, hydrobromic, sulfuric, acetic, maleic, succinic or methanesulfonic. If the final compound contains an acidic group, cationic salts can be prepared. Typically the main compound is treated with an excess of an alkaline reagent, such as hydroxide, carbonate or alkoxide, which contains the appropriate cation. Cations such as Na +, K ', Ca + 2 and NH 4 * are examples of cations present in pharmaceutically acceptable salts. Certain of the compounds form inner salts or zwitterions that may also be appropriate.

The compounds described above are useful for the treatment of cell proliferation disorders, infectious diseases and immunological diseases in mammals, and specifically, in human patients requiring such treatment. The cell proliferative disorders that can be treated using the composition described above include cancer, cardiovascular disease such as myocarditis and restenosis after angioplasty, kidney diseases such as lupus and polycystic kidney disease, rejection of host graft, gout and other disorders. proliferative Autoimmune diseases that can be treated with the compositions described above include rheumatic arthritis, lupus, type I diabetes, multiple sclerosis, and similar disorders and diseases. Infectious diseases that can be treated using the compositions described above include D3D, Crohn's disease, AIDS, ARDS and similar disorders. The compositions described above can also be used to treat fungal infections, dermatological diseases such as psoriasis, abnormal wound healing, keloids, immunological diseases such as autoimmunity, asthma, allergies, acute and delayed hypersensitivity, graft-versus-host disease and neuroimmune disease such as multiple sclerosis and acute disseminated encephalomyelitis.

The method of treating these diseases and disorders comprises parenterally or orally administering an effective amount of the chosen compound or combinations thereof, preferably dispersed in a pharmaceutical carrier. The unit doses of the active ingredient are generally chosen in the range of 0.01 to 100 mg / kg, but will be readily determined by one skilled in the art depending on the route of administration, age and condition of the patient. These unit doses can be administered one to ten times daily for acute or chronic diseases. No unacceptable toxicological effect is expected when the compounds of the invention are administered according to the present invention.

The pharmaceutical compositions of the compounds of this invention, or derivatives thereof, can be formulated as lyophilized solutions or powders for parenteral administration. The powders can be reconstituted by the addition of an appropriate diluent to another pharmaceutically acceptable carrier before use. The liquid formulation is usually a regulated, isotonic aqueous solution. Examples of the appropriate diluents are standard isotonic saline solution, dextrose at normal% in water or regulated sodium solution or ammonium acetate. Said formulation is especially suitable for parenteral administration, but can also be used for oral administration. It may be desirable to add excipients such as polyvinylpyrrolidone, gelatin, hydroxycellulose, acacia, polyethylene glycol, mannitol, sodium chloride or sodium citrate. As an alternative, these compounds can be encapsulated, tabletted or prepared in an emulsion or syrup for oral administration. The pharmaceutically acceptable solid or liquid carriers can be added to improve or stabilize the composition, or to facilitate the preparation of the composition. Liquid carriers include syrup, peanut oil, olive oil, saline, alcohols, and water. Solid carriers include starch, lactose, calcium sulfate, dehydrate, terra alba, magnesium stearate or stearic acid, talc, pectin, acacia, agar or gelatin. The carrier can also include a sustained release material such as glyceryl monostearate or glyceryl distearate, alone or with a wax. The amount of the solid carrier varies but, preferably, it will be between about 20 mg to about 1 g per unit dose. The pharmaceutical preparations are made according to conventional pharmacy techniques which include milling, mixing, granulation and compression, when necessary, for the tablet forms; or ground, mixed and stuffed for hard gelatin capsule forms. When a liquid carrier is used, the preparation will take the form of a syrup, elixir, emulsion or an aqueous or non-aqueous suspension. Such a liquid formulation can be directly administered p.o. or fill in a soft gelatin capsule.

EXAMPLE 1 The compounds useful in the therapeutic method of this invention are prepared by conventional methods of organic chemistry. Coupling reagents are well known in the art, such as DCC or other carbodiimides, EDC, BOP and PPA, and can optionally be used with other reagents, such as HOBT, NMM and DMAP, which can facilitate the reaction. The preparation of the compounds of Formula (1) wherein Di, D2 and E are amino acids is well known in the art using any phase techniques in solution or solid phase as described by Bodanszky, "The Practice of Peptide Synthesis" , Springer - Verlag, First Edition, 1984. The appropriate protective groups for the amino group are those presented by Greene, et al., "Protective Group in Organic Synthesis ", Second Edition, John Wiley and Sons, New York, New York, 1991. The benzyloxycarbonyl, t-butoxycarbonyl and fluorenylmethoxycarbonyl groups are especially useful amino protecting groups.

The synthesis of peptides in solid phase was achieved as follows: Rink amide resin is placed in a syringe coupled with a fragmented filter. The resin is deprotected using 20% piperidine in DMF. After 20 minutes, the resin is washed five times with DMF, five times with methanol, then five times with DMF. A solution of amino acid (E), carbodiimide and HOBT in DMF is removed in the syringe and the reaction mixture is allowed to mix for 3 to 20 hours. The reaction solution is expelled and the mixture is washed five times with DMF, five times with methanol, then five times with DMF. This cycle was repeated until the desired sequence was coupled. The final coupling used, 5-methoxy-1-indanone-3-acetic acid, carbodiimide and HOBT. After the final washings, the peptide fragment was divided from the resin using 95% TFA / 5% water. The concentration of the division mixture produces a white solid.

EXAMPLE 2 The compounds of this invention prepared according to the method of Example 1 were tested as follows. The 20S catalytic subunit of the proteasome (also known as the multicatalytic proteinase complex) was purified to homogeneity from bovine brain according to published methods (Wil, S. and Orlowski, M., 40 842 J. Neurochem., (1983 )). The chymotryptic activity of the complex was measured in fluorescence after cleavage of the substrate peptide succinyl-leucine-leucine-valine-tyrosine-7-amino-4-methyl coumarin. The standard in vitro assay consists of 2 μg of proteasome 20S, 0.1 - 100 μg / ml protease inhibitor in 200 μl of 50 nM HEPES, containing 0. 1% sodium dodecyl sulfate, pH 7.5. The proteolytic reaction is initiated by the addition of 50 mM fluorogenic peptide substrate and allowed to progress for 15 minutes at 37 ° C. The reaction was terminated by the addition of 100 μl of lOOmM acetate buffer, pH 4.0. The proportion of proteolysis is directly proportional to the amount of aminomethylcoumarin released which is measured by fluorescence spectroscopy (EX 370 nm, EM 430 nm). The structures of the tested compounds as well as the results of the tests are reported in Table 2, below.

TABLE 2 EXAMPLE 3 The compounds prepared according to the method of Example 1 were tested against several different cell lines. Cell monolayers were cultured in the presence of the compound tested for 18 hours to assess their ability to inhibit cell proliferation. Cell proliferation was determined colorimetrically using the Celltiter 96 aqueous non-radioactive cell proliferation assay (Promega) wherein the cell proliferation is directly proportional to the absorbance at 490 nm. The results are cited as IC5o in μg / ml for the inhibition of cell proliferation in several cell types.

EXAMPLE 4 The compounds prepared according to the method of Example 1 were tested by the inhibition of TNS synthesis induced by LPS. The RAW cells were pretreated with different concentrations of test compound for 1 hour before the administration of lipopolysaccharide (100 ng / ml). Cell culture supernatants were collected after 1 hour and assayed for TNF concentration by ELISA (Biosource).

EXAMPLE 5 This example examines the ability of compound 173 and particularly compound 187 described in Tables 1 and 2 above to inhibit proteasome activity as indicated, in part, by the presence of IβB and / or pl05 in inhibited cells. . For NF-KB to be transferred to the nucleus in response to a stimulus such as lipopolysaccharide (LPS) and activate transcription, two proteolytic events need to occur, namely the degradation of the inhibitory protein I B and the processing of p 105 to p50. These proteolytic events serve to unmask the localization signal of NF-KB.

Inhibition of I? B Degradation induced by LPS The RAW cells were pretreated with different concentrations of the test compound for 1 hour before the administration of lipopolysaccharide (100 ng / ml). The whole cell lysates were cultured after 1 hour, 10 μg of protein were separated by SDS-PAGE, transferred to nitrocellulose and assayed by immunoreactivity with anti-I? B antibody. Western blots (see Figure 1) were visualized using the Boehringer Manheim chemiluminescent detection kit. The blot shows that I? B is present in the treated cells with as little as 5 μg / ml of compounds 173 and 187.

Inhibition of Processing of pl05 to p50 induced by LPS Compound 187 as described in Tables 1 and 2 above was used to pretreat RAW cells as described above, and whole cell lysates prepared as described above were analyzed by immunoreactivity to an antibody anti p50. The results, set forth in Figure 2 indicate that p50 and pl05 are both present in the treated cells with as little as 5 μg / ml of compound 187, whereas in the untreated cells, most of pl05 has been processed to p50. .

Inhibition of Transfer of NF-KB to the Nuclear Fraction of the Cell Induced by LPS RAW cells were treated for 1 hour with compound 187 (20 μg / ml) and then incubated with lipopolysaccharide (100 ng (ml) for an additional hour Nuclear fractions were prepared according to standard procedures Link reactions for gel mobility change assays contained 5 μg of protein core extract, 50,000 cpm of NF-KB consensus binding oligonucleotide labeled 32 P in the presence and the absence of an excess of fifty unlicensed oligonucleotide moieties, the gel mobility change test, established in the Figure 3 shows that compound 187 is effective to inhibit the accumulation of NF-K in the nucleus of cells.

Claims (58)

CLAIMS: 1. A method and compositions for treating mammals with disorders of cell proliferation, disorders of the immune system and infectious diseases comprising administering to the mammal a therapeutically effective amount of a compound having the formula: wherein Ri-Ri are each individually selected from the group including hydrogen, halogen, hydroxyl, thiol, lower alkyl, substituted lower alkyl, alkenyl, alkynyl, alkyl alkenyl, alkyl alkynyl, alkoxy, alkylthio, acyl, aryloxy, amino, amido, carboxyl, aryl, substituted aryl, heterocycle, heteroaryl, substituted heterocycle, heteroalkyl, cycloalkyl, substituted cycloalkyl, alkylcycloalkyl, alkylcycloheteroalkyl, nitro or cyano.
1. R5-R9 are each independently selected from the group of compounds including hydrogen, halogen, hydroxyl, thiol, oxo, lower alkyl, substituted lower alkyl, alkenyl, alkynyl, alkyl alkenyl, alkyl alkynyl, alkoxy, alkylthio, acyl, aryloxy, amino, amido, carboxyl, aryl, substituted aryl, heterocycle, heteroaryl, substituted heterocycle, heteroalkyl, cycloalkyl, substituted cycloalkyl, alkylcycloalkyl, alkylcycloheteroalkyl, nitro or cyano; "- - X is selected from the group of compounds including hydrogen, - Di, - D2, - E, - Di - E, D2 - E, - Di - D2 or a compound having the formula: ^ - wherein Di and D2 are each independently chosen from the group of compounds that include a compound having the formula hydrogen, halogen, thiol, lower alkyl, substituted minor alkyl, alkenyl, alkynyl, alkylalkenyl, alkyl alkynyl, alkoxy, alkylthio, acyl, aryloxy, amino, amido, carboxyl, aryl, substituted aryl, heterocycle, heteroaryl, substituted heterocycle, heteroalkyl, cycloalkyl, substituted cycloalkyl, alkylcycloalkyl, alkylcycloheteroalkyl; wherein E is a compound selected from the group that includes: hydrogen, halogen, thiol, lower alkyl, substituted minor alkyl, alkenyl, alkynyl, alkylalkenyl, alkyl alkynyl, alkoxy, alkylthio, acyl, aryloxy, amino, amido, carboxyl, aryl, substituted aryl, heterocycle, heteroaryl, substituted heterocycle, heteroalkyl, cycloalkyl, substituted cycloalkyl, alkylcycloalkyl, alkylcycloheteroalkyl; where Rio - R? are each independently selected from the group of compounds including hydrogen, halogen, hydroxyl, thiol, oxo, lower alkyl, substituted lower alkyl, alkenyl, alkynyl, alkyl alkenyl, alkyl alkynyl, alkoxy, alkylthio, acyl, aryloxy, amino, amido , carboxyl, aryl, substituted aryl, heterocycle, heteroaryl, substituted heterocycle, heteroalkyl, cycloalkyl, substituted cycloalkyl, alkylcycloalkyl, alkylcycloheteroalkyl, nitro or cyano; Ji and J2 are N-R15, CR? 6 RJ7, O, S- (O) 0.2, P- (O) 0.3; and R15-Rp are each independently selected from the group of compounds including hydrogen, halogen, hydroxyl, thiol, oxo, lower alkyl, substituted lower alkyl, alkenyl, alkynyl, alkyl alkenyl, alkyl alkynyl, alkoxy, alkylthio, acyl, aryloxy , amino, amido, carboxyl, aryl, substituted aryl, heterocycle, heteroaryl, substituted heterocycle, heteroalkyl, cycloalkyl, substituted cycloalkyl, alkylcycloalkyl, alkylcycloheteroalkyl or cyano.
2. 2. The method and compositions according to Claim 1 wherein Ri-R4 are each individually selected from the group including hydrogen, halogen, lower alkyl, substituted lower alkyl, aryl, aryloxy, substituted aryl, amino, amido, alkoxy, thio, alkyl thiol, hydroxyl, cyano, nitro, acyl, carboxyl and alkynyl and R5-R9 are individually selected from the group consisting of hydrogen, lower alkyl, substituted lower alkyl, aryl, substituted aryl, amino, amido, alkoxy and alkynyl.
3. 3. The method and compositions according to Claim 1 wherein X is selected from the group consisting of - Di - E, or - D2 - E.
4. 4. The method and compositions according to Claim 3 wherein -DI and -D2 are each:
5. 5. The method and compositions according to Claim 4 wherein Ji is N-R15, and Rs-R11 and R15 are each individually selected from the group of compounds consisting of hydrogen, lower alkyl, substituted lower alkyl, aryl, and substituted aryl and E is selected from the group consisting of lower alkyl, substituted lower alkyl, aryl, substituted aryl, alkoxy and amino.
6. 6. The method and compositions according to Claim 5 wherein R5-Rp and R15 are individually selected from the group consisting of hydrogen, lower alkyl, substituted lower alkyl and E is selected from the group consisting of lower alkyl, substituted lower alkyl , aryl, substituted aryl, alkoxy and amino.
7. 7. The method and compositions according to Claim 6 wherein E is selected from the group consisting of alkoxy and amino, and Ri-R- * are selected from the group consisting of hydrogen, halogen, lower alkyl, substituted lower alkyl, alkoxy, amino, nitro, hydroxyl, cyano, alkynyl, thio and alkylthio.
8. 8. The method and compositions according to Claim 7 wherein E is NR'R "wherein R 'and R" are each individually selected from the group consisting of hydrogen, lower alkyl, substituted lower alkyl, aryl, substituted aryl and cycloalkyl and Ri-R4 are each individually selected from the group consisting of hydrogen, halogen, nitro, lower alkyl, substituted minor alkyl, amino, alkylthio, thio, hydroxy and alkoxy.
9. 9. The method and compositions according to Claim 8 wherein Ri-R4 are each individually selected from the group consisting of hydrogen, alkoxy, and lower alkyl, R5-Rn and Ru are each selected individually from the group it consists of hydrogen, lower alkyl, and substituted lower alkyl, and R 'and R "are each individually selected from the group consisting of hydrogen, lower alkyl, substituted minor alkyl, and cycloalkyl.
10. 10. The method and compositions according to Claim 2, wherein X is: Q- //
11. 11. The method and compositions according to Claim 10, wherein i and -D2 are each:
12. 12. The method and compositions according to Claim 11 wherein E is a compound selected from the group consisting of hydrogen, lower alkyl, substituted minor alkyl, aryl, substituted aryl, amino, cycloalkyl, substituted cycloalkyl, alkoxy or alkylcycloalkyl.
13. 13. The method according to Claim 11 wherein E is:
14. 14. The method according to Claim 13 wherein Ji and J2 are each N R15 wherein Rio-R13 and R15 are each individually selected from the group consisting of hydrogen, lower alkyl, substituted lower alkyl, aryl and substituted aryl and R? 4 is selected from the group consisting of lower alkyl, substituted lower alkyl , aryl, substituted aryl, amino and alkoxy.
15. 15. The method according to Claim 14 wherein R 5 -R 3 are each selected individually from the group consisting of hydrogen, lower alkyl, substituted minor alkyl, aryl and substituted aryl, R 4 is selected from the group of compounds that consists of alkoxy and amino, and R15 is selected from the group of compounds consisting of hydrogen, lower alkyl, substituted lower alkyl, aryl and substituted aryl.
16. 16. The method according to Claim 15 wherein Ri-R »are each individually selected from the group of compounds consisting of hydrogen, halogen, lower alkyl, substituted lower alkyl, aryl, aryloxy, aryl substituted, amino, alkylthio, nitro , hydroxy, thio and alkoxy.
17. 17. The method according to Claim 16 wherein R5-R13 and R15 are each selected from the group consisting of hydrogen, lower alkyl, and substituted lower alkyl.
18. 18. The method according to Claim 17 wherein R? is NR'R "wherein R 'and R" are each individually selected from the group of compounds consisting of hydrogen, lower alkyl, substituted lower alkyl, aryl, substituted aryl, and cycloalkyl.
19. 19. The method according to Claim 18 wherein Ri-R-4 are each individually selected from the group consisting of hydrogen, alkoxy, and lower alkyl, R5 -R13 and R15 are each selected individually from the group of compounds which consists of hydrogen, lower alkyl, and substituted lower alkyl, and R 'and R "are each individually selected from the group of compounds consisting of hydrogen, minor alkyl, cycloalkyl, and substituted minor alkyl.
20. 20. The method according to Claim 12 wherein Ji is -N R15 wherein R15 is selected from the group consisting of hydrogen, lower alkyl, substituted lower alkyl, aryl and substituted aryl.
21. 21. The method according to Claim 20 wherein R5 - Rn are each individually selected from the group consisting of hydrogen, lower alkyl, substituted lower alkyl, aryl and substituted aryl, E is selected from the alkoxy and amino, Ri - R_? each is individually selected from the group of compounds consisting of hydrogen, halogen, lower alkyl, substituted lower alkyl, alkyl thio, thio, alkoxy, amino, nitro and hydroxyl, and R15 is a compound selected from the group of compounds consisting of of hydrogen, minor alkyl and aryl.
22. 22. The method according to Claim 21 wherein E is N'R "wherein R 'and R" are each individually selected from the group of compounds consisting of hydrogen, lower alkyl, substituted lower alkyl, aryl, aryl substituted and cycloalkyl.
23. 23. The method according to Claim 21 wherein Ri - Rj are each individually selected from the group consisting of hydrogen, alkoxy, and lower alkyl, R5 - Rp are each individually selected from the group consisting of hydrogen, alkyl minor, substituted lower alkyl, R 'and R "are each individually selected from the group consisting of hydrogen, lower alkyl, substituted lower alkyl and cycloalkyl, and R15 is selected from the group of compounds consisting of hydrogen and lower alkyl.
24. 24. The method according to Claim 10 wherein R3 is methoxy, -Di is leucine and -D2 is leucine and E is NR'R ".
25. 25. The method according to Claim 24 wherein - Di is 1 - leucine and - D2 is d - leucine.
26. 26. The method according to claim 25 wherein E is selected from the group of compounds consisting of benzylamine, 1-indanylamine, N, N'-dibenzylamine, 2,6-difluorobenzylamine, 4-methoxybenzylamine, piperonyl amine and N? 2.
27. 27. The method according to Claim 24 wherein E is glycinamide.
28. 28. The method according to Claim 1 wherein the mammal is a human.
29. 29. The method according to claim 1 wherein the therapeutically effective amount ranges from about 0.001 to about 100 mg / kg of mammalian weight.
30. 30. The method of Claim 1 wherein the composition is administered to a mammal suffering from a cell proliferation disorder selected from the group consisting of rheumatic arthritis, lupus, type I diabetes, multiple sclerosis, cancer, restenosis, graft host disease. and drop.
31. 31. The method of Claim 1 wherein the cell proliferation disorder is restenosis.
32. 32. The method of Claim 30 wherein the cell proliferation disorder is cancer.
33. 33. The method of Claim 30 wherein the therapeutic agent induces apoptosis.
34. 34. The method of Claim 30 wherein the cell proliferation disorder is polycystic kidney disease.
35. 35. The method of Claim 1 wherein the composition is administered to a mammal suffering from an infectious disease.
36. 36. The method according to claim 35 wherein the infectious disease is selected from the group consisting of L3D, Crohn's disease, AIDS, ARDS and fungal diseases.
37. 37. The method according to claim 1 wherein the composition is administered to a mammal suffering from an immunological disease such as rheumatic arthritis, immunological diseases, rejection of transplants and psoriasis.
38. 38. The therapeutically effective amount of the composition useful in the method of Claim 1 wherein the composition is administered in the form of a solution.
39. 39. The therapeutically effective amount of the composition useful in the method of Claim 1 wherein the composition is administered in the form of a tablet.
40. 40. Compositions that have the formula: Where R1-R9 are each individually selected from the group consisting of hydrogen, halogen, lower alkyl, substituted lower alkyl, alkoxy, amino, nitro, hydroxyl, cyano, alkynyl, thio, and alkylthio. X is a compound that has the formula: Where Di and D2 are each individually: Wherein i is N-R15, wherein Rio, Rp, are each individually selected from the group of compounds consisting of hydrogen, lower alkyl, substituted lower alkyl, aryl and substituted aryl and E is selected from the group consisting of lower alkyl , substituted lower alkyl, aryl, substituted aryl, alkoxy, amino, amino acid, and NR'R "wherein h is selected from the group of compounds including N-R15, - CR16 Rp, O, S- (0) or-2, P- (0) 0-3, wherein Rp-Rp may each individually chosen from the group of compounds including hydrogen, halogen, hydroxyl, oxo, thiol, lower alkyl, substituted lower alkyl, alkynyl, alkylalkenyl, alkyl alkynyl, alkoxy, alkylthio, acyl, aryloxy, amino, amido, carboxyl, aryl , substituted aryl, heterocyclic, heteroaryl, substituted heterocycle, heteroalkyl, cycloalkyl, substituted cycloalkyl, alkylcycloalkyl and wherein R 'and R "are each selected from the group consisting of hydrogen, lower alkyl, substituted lower alkyl, aryl, aryl substituted and cycloalkyl.
41. 41. The compositions of claim 40 wherein R1-R9 are each individually selected from the group consisting of hydrogen, halogen, nitro, lower alkyl, substituted lower alkyl, amino, alkylthio, thio, hydroxy and alkoxy.
42. 42. The compositions of claim 40 wherein R? -R2 and R-^ -Rg are each hydrogen.
43. 43. The compositions of claim 40 wherein R3 is -O-CH3.
44. 44. The compositions of claim 40 wherein Rio-Rn and R15 are each individually selected from the group consisting of hydrogen, lower alkyl and substituted lower alkyl and E is selected from the group consisting of lower alkyl, substituted lower alkyl, aryl, aryl substituted, alkoxy and amino acid.
45. 45. The compositions of claim 40 wherein Ji is NR'R "wherein R 'and R" are each individually selected from the group consisting of hydrogen, lower alkyl, substituted lower alkyl and cycloalkyl.
46. 46. The compositions of claim 40 wherein E is a compound selected from the group consisting of hydrogen, lower alkyl, substituted minor alkyl, aryl, substituted aryl, amino, amino acid, cycloalkyl, substituted cycloalkyl, alkoxy or alkylcycloalkyl.
47. 47. The compositions of claim 40 wherein E is amino acid, or NR'R "wherein R 'and R" are each individually selected from the group of compounds consisting of hydrogen, lower alkyl, substituted lower alkyl, aryl, aryl substituted and cycloalkyl.
48. 48. The compositions of claim 42 wherein Ji and J2 are each -N-R15 wherein Rio-R13 and Ru are each individually selected from the group consisting of hydrogen, lower alkyl, substituted lower alkyl, aryl and substituted aryl and R 4 is selected from the group consisting of lower alkyl, substituted lower alkyl, aryl, substituted aryl, amino and alkoxy.
49. 49. The compositions of claim 40 wherein R5-R13 are each individually selected from the group consisting of hydrogen, lower alkyl, substituted lower alkyl, aryl and substituted aryl, R? is selected from the group of compounds consisting of alkoxy and amino and R15 is selected from the group of compounds consisting of hydrogen, lower alkyl, substituted lower alkyl, aryl and substituted aryl.
50. 50. The compositions that have the formula: Where R? -R2 and R-t-R9 are each hydrogen, and R3 is -OCH3; X is D, l_ D 2 Where Di and D2 are each amino acids and E is an amino acid or -N-R15 wherein R15 is selected from the group of compounds selected from hydrogen, lower alkyl, substituted lower alkyl, aryl, substituted aryl and aralkyl.
51. 51. The compounds of claim 50 wherein Di and D2 are each selected from -leu or D-leu.
52. 52. The compounds of claim 50 wherein E is selected from gli-NH2, hydrogen, benzyl, dibenzyl, indane, n-benzylhydroxamine, 2,6-difluorobenzyl and piperonyl.
53. 53. Compositions that have the formula: Where Ri is isopropyl, benzyl, hydroxyethyl, adamantyl, phenyl, phenethyl or cyclohexylmethyl and R2 is hydrogen or methyl.
54. 54. The compounds of claim 53 wherein Ri is benzyl and R2 is hydrogen.
55. 55. The compositions of claim 53 wherein Ri and R2 are each selected from the group including hydrogen, 1-indanyl, piperonyl, 2-6-difluorobenzyl, indane, benzyl, dibenzyl, 4-methoxybenzyl, 4-nitrobenzyl, pentafluorobenzyl, furfuryl , diphenylmentil, 4-phenylbenzyl, 2-phenylbenzyl, 4-benzyloxy and benzoftirfuril.
56. 56. Compositions that have the formula:
57. 57. The compositions of claim 56 wherein R is 1 -indanyl, piperonyl, 2-6-difluorobenzyl, indane, benzyl, dibenzyl, 4-methoxybenzyl, 4-nitrobenzyl, pentafluorobenzyl, furfuryl, diphenylmethyl, 4-phenylbenzyl, 2-phenylbenzyl, 4- benzyloxybenzyl and benzofurfuryl.
58. 58. The compositions of claim 56 wherein R is 2,6-difluorobenzyl. EXTRACT OF THE INVENTION This invention is a method to inhibit cell proliferation using indanones.