JP2009513644A - Therapeutic compositions and methods - Google Patents

Abstract

The present invention relates to novel compounds, methods for their discovery, and therapeutic uses thereof. In particular, the present invention provides benzodiazepine derivatives and structural and functional related compounds, and methods of using benzodiazepine derivatives and related compounds as therapeutic agents for treating a number of conditions.

Description

  This application claims priority to US Provisional Application No. 60 / 730,711, filed Oct. 26, 2005, which is hereby incorporated by reference in its entirety.

  This invention was made with government support under grant AI47450 awarded by the National Institutes of Health. The government has certain rights in the invention.

The present invention relates to novel compounds, methods for their discovery, and therapeutic uses thereof. In particular, the present invention provides benzodiazepine derivatives and structural and functional related compounds and methods of using benzodiazepine derivatives and related compounds as therapeutic agents for treating a number of conditions.

BACKGROUND OF THE INVENTION Multicellular organisms accurately control cell numbers. The balance between cell growth and cell death achieves this homeostasis. Cell death occurs in almost all types of vertebrate cells via necrosis or by a suicide form of cell death known as apoptosis. Apoptosis is triggered by a variety of extracellular and intracellular signals involved in a common genetically programmed death mechanism.

  Multicellular organisms use apoptosis to instruct damaged or unnecessary cells to destroy themselves for the organism. Therefore, the control of the apoptotic process is very important for normal development, for example, the development of the embryonic period of the finger, and the toes require controlled removal of excess interconnected tissue due to apoptosis, and the nerves in the brain The formation of synapses is so. Similarly, controlled apoptosis is involved in peeling off the inner uterine wall (endometrium) at the beginning of menstruation. While apoptosis plays an important role in tissue sculpting and normal cell maintenance, apoptosis also provides a primary defense against cells and invaders (eg, viruses) that threaten the health of the organism.

  As expected, many diseases are associated with abnormal regulation of the cell death process. Experimental models have established a causal relationship between abnormal apoptotic regulation and the pathogenesis of various neoplastic, autoimmune and viral diseases. For example, in a cell-mediated immune response, effector cells (eg, cytotoxic T lymphocytes “CTL”) destroy infected cells by inducing virus-infected cells to undergo apoptosis. The organism then relies on the apoptotic process to destroy the effector cells when they are no longer needed. Autoimmunity is usually blocked by CTLs that induce apoptosis to each other and to themselves. Defects in this process are associated with various autoimmune diseases such as lupus erythematosus and rheumatoid arthritis.

  Multicellular organisms also use apoptosis to direct cells with damaged nucleic acids (eg, DNA) to destroy themselves before becoming cancerous. Some oncogenic viruses overcome this protective measure by reprogramming infected (transformed) cells and disrupting the normal apoptotic process. For example, some human papillomaviruses (HPV) are involved in inducing cervical cancer by inhibiting the removal of apoptotic cells from transformed cells by producing a protein (E6) that inactivates the p53 apoptosis promoter is doing. Similarly, the causative agent of Epstein-Barr virus (EBV), mononucleosis and Burkitt lymphoma reprograms the infected cells to produce proteins that block the normal removal of apoptotic cells from the abnormal cells, and thus cancer cells Can grow and spread throughout the organism.

Yet another virus destructively manipulates cell apoptosis mechanisms without directly leading to cancer development. For example, the destruction of the immune system in individuals infected with human immunodeficiency virus (HIV) may proceed via infected CD4 ⁺ T cells (approximately 1 in 100,000) that direct uninfected sister cells to undergo apoptosis. It is done.

  Some cancers that arise by non-viral means have also developed mechanisms to escape destruction by apoptosis. For example, melanoma cells avoid apoptosis by inhibiting the expression of the gene encoding Apaf-1. Other cancer cells, particularly lung and colon cancer cells, secrete high levels of soluble decoy molecules that prevent the initiation of CTL-mediated clearance of abnormal cells. Dysregulation of apoptotic mechanisms is also implicated in various degenerative conditions and vascular diseases.

  It is clear that controlled regulation of the apoptotic process and its cellular mechanisms is essential for the survival of multicellular organisms. Typically, biochemical changes that occur within cells that are directed to undergo apoptosis occur in an ordered progression. However, as noted above, defective regulation of apoptosis can cause severe adverse effects in the organism.

  There are various attempts to control and restore the regulation of apoptotic mechanisms in abnormal cells (eg, cancer cells). For example, much research has been done to develop cytotoxic drugs that destroy abnormal cells before they proliferate. As such, cytotoxic drugs have expanded utility in both human and animal health, treating almost all types of cancer and hyperproliferative autoimmune diseases such as lupus erythematosus and rheumatoid arthritis. The first line is shown.

  Many cytotoxic drugs in clinical use are efficacious by damaging DNA (eg, cis-diaminodicloplatanim (II) cross-links to DNA, while bleomycin induces strand breaks). The result of this nuclear damage, when recognized by cellular factors such as the p53 system, initiates the apoptotic cascade leading to the death of damaged cells.

  However, existing cytotoxic chemotherapeutic drugs have significant drawbacks. For example, many known cytotoxic drugs are almost indistinguishable between healthy and abnormal cells. This lack of specificity often results in serious side effects that limit efficacy and / or can lead to premature death. In addition, long-term administration of many existing cytotoxic drugs results in the expression of resistance genes (e.g., bcl-2 family or multidrug resistance (MDR) proteins), and the administration is less effective or useless It becomes. Some cytotoxic drugs induce mutations in p53 and related proteins. In view of these, an ideal cytotoxic drug should kill only abnormal cells and be less susceptible to chemical resistance.

  One strategy to selectively kill abnormal cells or block their growth is the development of drugs that selectively recognize molecules expressed in abnormal cells. Thus, effective cytotoxic chemotherapeutic drugs recognize disease indicating molecules and induce abnormal cell death (eg, directly or indirectly). Although several types of cancer cell markers have been identified and targeted by therapeutic antibodies and small molecules, unique traits for diagnosis and therapeutic development are not known for most cancers. Furthermore, in diseases such as lupus, specific molecular targets for drug development have not been identified.

  What is needed is to regulate the apoptotic process in subjects suffering from diseases and conditions characterized by dysregulation of these processes (e.g., viral infections, hyperproliferative autoimmune diseases, chronic inflammatory conditions, and cancer). Improved compositions and methods.

Overview The present invention provides novel compounds used in the treatment of many diseases and conditions in humans and animals, and used in research, compound screening, research applications, and diagnostic applications. The present invention also provides the use of these novel compounds that elicit special biological responses, as well as the use of known compounds (e.g., compounds that bind to specific target molecules and / or cause special cellular events). To do. Such compounds and uses are described throughout this application and represent a diverse collection of compositions and uses.

  Certain preferred compositions and uses are described below. The present invention is not limited to these special compositions and uses. The present invention provides a number of useful compositions as described throughout this application.

  Certain embodiments of the invention include compositions comprising a benzodiazepine compound having a chemical moiety that causes the benzodiazepine to lack an asymmetric center associated with the third carbon position of the benzodiazepine ring.

RおよびS鏡像体の両方ならびにラセミ混合物が含まれる。 Certain embodiments of the invention include compositions comprising a benzodiazepine compound having a chemical moiety that causes the benzodiazepine to lack an asymmetric center associated with the third carbon position of the benzodiazepine ring. In some embodiments, the composition has the formula:

Both R and S enantiomers as well as racemic mixtures are included.

In some embodiments, A ---- B is selected from the group consisting of N-CH ₂ and C = N.

  In some embodiments, R5 is a linker group and is present or absent.

  In some embodiments, R1 is an isostere of OH. In some embodiments, R1 is hydrogen; halogen; OH; a chemical moiety comprising an aryl subgroup; a chemical moiety comprising a substituted aryl subgroup; a chemical moiety comprising a cycloaliphatic subgroup; a substituted cycloaliphatic subgroup Chemical moiety comprising a heterocyclic subgroup; chemical moiety comprising a substituted heterocyclic subgroup; chemical moiety comprising at least one ester subgroup; chemical moiety comprising at least one ether subgroup; linear or branched Selected from the group consisting of: a saturated or unsaturated, substituted or unsubstituted aliphatic chain having at least two carbons; a sulfur containing chemical moiety; a nitrogen containing chemical moiety; and —OR—, wherein R is Chemical moiety comprising an aryl subgroup; Chemical moiety comprising a substituted aryl subgroup; Chemical moiety comprising a cycloaliphatic subgroup; Chemical moiety comprising a substituted cycloaliphatic subgroup; Chemical moiety comprising a heterocyclic subgroup A chemical moiety comprising a substituted heterocyclic subgroup; an aliphatic chain having at least two carbons, linear or branched, saturated or unsaturated, substituted or unsubstituted; a chemical moiety comprising at least one ester subgroup; Selected from the group consisting of a chemical moiety comprising one ether subgroup; a chemical moiety comprising sulfur; and a chemical moiety comprising nitrogen.

式中、R1’は、ハロゲン；アルキル；置換アルキル；アリール；置換アリール；アミノ；カルボニル；スルホン；スルホンアミド；エーテル；OH；硫黄を含む化学部分；窒素を含む化学部分；CH₃；直鎖または分枝、飽和または不飽和の少なくとも1つの炭素を有する脂肪族鎖；直鎖または分枝、飽和または不飽和の少なくとも2つの炭素を有し、少なくとも1つのヒドロキシサブ基を有する脂肪族鎖；直鎖または分枝、飽和または不飽和の少なくとも2つの炭素を有し、少なくとも1つのチオールサブ基を有する脂肪族鎖；直鎖または分枝、飽和または不飽和の少なくとも2つの炭素を有し、アルデヒドサブ基で終わる脂肪族鎖；直鎖または分枝、飽和または不飽和の少なくとも2つの炭素を有し、少なくとも1つのケトンサブ基を有する脂肪族鎖；直鎖または分枝、飽和または不飽和の少なくとも2つの炭素を有し、カルボン酸サブ基で終わる脂肪族鎖；直鎖または分枝、飽和または不飽和の少なくとも2つの炭素を有し、少なくとも1つのアミドサブ基を有する脂肪族鎖；直鎖または分枝、飽和または不飽和の少なくとも2つの炭素を有し、少なくとも1つのアシル基を有する脂肪族鎖；直鎖または分枝、飽和または不飽和の少なくとも2つの炭素を有し、少なくとも1つの窒素含有部分を有する脂肪族鎖；直鎖または分枝、飽和または不飽和の少なくとも2つの炭素を有し、少なくとも1つのアミンサブ基を有する脂肪族鎖；直鎖または分枝、飽和または不飽和の少なくとも2つの炭素を有し、少なくとも1つのエーテルサブ基を有する脂肪族鎖；直鎖または分枝、飽和または不飽和の少なくとも2つの炭素を有し、少なくとも1つのハロゲンサブ基を有する脂肪族鎖；直鎖または分枝、飽和または不飽和の少なくとも2つの炭素を有し、少なくとも1つのニトロニウムサブ基を有する脂肪族鎖からなる群より選択される。 In some embodiments, R1 is selected from the group consisting of:

Wherein, R1 'is halogen, alkyl, substituted alkyl, aryl, substituted aryl; amino; carbonyl; sulfone; sulfonamide; ethers; OH; chemical moieties which contain a nitrogen; chemical moiety containing a sulfur CH _3; straight or An aliphatic chain having at least one carbon that is branched, saturated or unsaturated; an aliphatic chain having at least two carbons that is linear or branched, saturated or unsaturated, and having at least one hydroxy subgroup; An aliphatic chain having at least two carbons that are chain or branched, saturated or unsaturated, and having at least one thiol subgroup; having at least two carbons that are linear or branched, saturated or unsaturated, and having an aldehyde sub Aliphatic chain ending in a group; straight chain or branched, saturated or unsaturated, aliphatic chain having at least one ketone subgroup; straight chain Or an aliphatic chain having at least two branched, saturated or unsaturated carbons and ending with a carboxylic acid subgroup; having at least two carbons that are straight or branched, saturated or unsaturated, and at least one An aliphatic chain having an amide subgroup; an aliphatic chain having at least two carbons that are linear or branched, saturated or unsaturated, and having at least one acyl group; at least one of linear or branched, saturated or unsaturated An aliphatic chain having two carbons and having at least one nitrogen-containing moiety; an aliphatic chain having at least two carbons that are linear or branched, saturated or unsaturated, and having at least one amine subgroup; An aliphatic chain having at least two carbons that are chain or branched, saturated or unsaturated, and having at least one ether subgroup; at least two carbons that are linear or branched, saturated or unsaturated An aliphatic chain having at least one halogen subgroup; a group consisting of an aliphatic chain having at least two carbons, straight or branched, saturated or unsaturated, and having at least one nitronium subgroup More selected.

  In some embodiments, R2 is a chemical moiety comprising an aryl subgroup; a chemical moiety comprising a substituted aryl subgroup; a chemical moiety comprising a cycloaliphatic subgroup; a chemical moiety comprising a substituted cycloaliphatic subgroup; a heterocyclic subgroup A chemical moiety comprising a substituted heterocyclic subgroup; a chemical moiety comprising sulfur; and a chemical moiety comprising nitrogen. In some embodiments, R2 is a cyclic group larger than benzene, where greater than benzene includes any chemical group containing 7 or more non-hydrogen atoms.

；
キノリン、および全ての芳香族位置異性体からなる群より選択される。 In some embodiments, R2 is naphthalene; phenol; 1-naphthalenol; 2-naphthalenol;

;
Selected from the group consisting of quinoline and all aromatic positional isomers.

  In some embodiments, R3 is an isostere of OH. In some embodiments, R3 is hydrogen; halogen; OH; a chemical moiety comprising at least one ester subgroup; a chemical moiety comprising at least one ether subgroup; linear or branched, saturated or unsaturated, substituted or An aliphatic chain having at least two unsubstituted carbons; a chemical moiety containing sulfur; a chemical moiety containing nitrogen; and -OR-, wherein R is a chemical moiety containing an aryl subgroup; A chemical moiety comprising an aryl subgroup; a chemical moiety comprising a cycloaliphatic subgroup; a chemical moiety comprising a substituted cycloaliphatic subgroup; a chemical moiety comprising a heterocyclic subgroup; a chemical moiety comprising a substituted heterocyclic subgroup; Branched, saturated or unsaturated, substituted or unsubstituted aliphatic chain having at least two carbons; at least one ester sub-group containing a chemical moiety; at least one ether sub-group containing It is selected from the group consisting of chemical moiety comprising nitrogen; chemical moiety comprising sulfur; chemical moiety.

In some embodiments, R3 is alkyl; mono-substituted alkyl; disubstituted alkyl; trisubstituted alkyl; (CH _{2) n,} wherein _{n = 1~6; CN; N 3} ; CNO; NH 2; SH; CF _{3; OCH 3; NCH 2 CH} (CH 2) N (CH 3) 2; NCH 2 CHCH 2 N (CH 3) 2; phenyl; 2-pyridyl; 3-pyridyl; 4-pyridyl; NCH _3; NCONHCH _{3; CH 2 OH; NHCONH 2; NHCOCH} 3; NHSO 2 CH 3; NHCN; NHCHO; SOCH 3; SO 2 CH 3; CHNOH; CHNOCH 3; SCH 3; CH 2 CO; CH 2 SO 2; CONH; CH 2 C ( _{NOH); CH 2 C (nOMe} ); NHSO 2 PH; NHCS; CH 2 NHCO; is selected from the group consisting of and NHCOS; COCH _2; NHCO _2. In some embodiments, R3 is an isotope described, for example, in Patani, G. and LaVoie, EJ, 1996, Chem. Rev. 96: 3147-3176, which is incorporated herein by reference in its entirety. Either is explained.

  In some embodiments, R3 is a cyclic structure attached at the 6,7 carbon position of the benzodiazepine structure, the 7,8 carbon position of the benzodiazepine structure, or the 8,9 carbon position of the benzodiazepine structure. In some embodiments, the cyclic structure comprises a chemical moiety comprising an aryl subgroup; a chemical moiety comprising a substituted aryl subgroup; a chemical moiety comprising a cycloaliphatic subgroup; a chemical moiety comprising a substituted cycloaliphatic subgroup; a heterocyclic subgroup A chemical moiety containing a substituted heterocyclic subgroup; a chemical moiety containing sulfur; a chemical moiety containing oxygen; and a chemical moiety containing nitrogen.

  In some embodiments, at least one of R1 and R3 is a chemical moiety that participates in hydrogen bonding. In some embodiments, the distance between the chemical moiety involved in hydrogen bonding and the R2 group in three-dimensional space differs, for example, by only about 12 angstroms.

であり、ここで、R4’は、直鎖または分枝、飽和または不飽和、置換または非置換の少なくとも2つの炭素を有する脂肪族鎖である。 In some embodiments, R4 is a chemical moiety that causes the benzodiazepine to lack an asymmetric center. In some embodiments, R4 is hydrogen,

Where R 4 ′ is a linear or branched, saturated or unsaturated, substituted or unsubstituted aliphatic chain having at least two carbons.

In some embodiments, the composition comprises the following chemical formula:

In some embodiments, the composition comprises a formula selected from the group consisting of:

であり；式中、R4''は1〜6個の炭素であり、そのいずれか1つは置換されるか、または非置換である。置換炭素としては、OH；ハロゲン；CH₃；直鎖または分枝、環状または非環状、飽和または不飽和の少なくとも2つの炭素を有する脂肪族鎖；ハロゲンを含む化学部分；硫黄を含む化学部分；窒素を含む化学部分；芳香族化学部分；親水性化学部分；および疎水性化学部分で置換されたものが挙げられるが、それらに限定されない。 In some embodiments, the structure is

Wherein R 4 ″ is 1 to 6 carbons, any one of which is substituted or unsubstituted. Substituted carbons include: OH; halogen; CH ₃ ; linear or branched, cyclic or acyclic, saturated or unsaturated aliphatic chain having at least two carbons; halogen containing chemical moiety; sulfur containing chemical moiety; Chemical moieties including nitrogen; aromatic chemical moieties; hydrophilic chemical moieties; and those substituted with hydrophobic chemical moieties include, but are not limited to.

In some embodiments, the compound is selected from the group consisting of:

In some embodiments, the compound is selected from the group consisting of:

  In one aspect, the present invention provides a method of treating cells with a composition of the present invention. Such methods are used in research, drug screening, and therapeutic applications. In some embodiments, the treatment is selected from the group consisting of inducing cell growth arrest in the target cell, inducing cell death in the target cell, and inducing cell apoptosis in the target cell. In some embodiments, the target cell is present in a subject having, for example, an autoimmune disease, a hyperproliferative disorder, an epidermal hyperplasia disorder, a pigmentation disorder, a cardiovascular disorder, and / or a viral disorder.

  In some embodiments, the target cell is selected from the group consisting of an in vitro cell, an in vivo cell, and an ex vivo cell. In another preferred embodiment, the target cell is a cancer cell. In yet another embodiment, the target cell is selected from the group consisting of B cells, T cells and granulocytes.

  In certain embodiments, the present invention provides a pharmaceutical composition comprising an agent selected from the following group: resveratrol, picetanol, estrogen, lansoprazole; and the benzodiazepine derivatives or structural and functional described in the present invention. Related compounds.

  In one aspect, the present invention provides a pharmaceutical composition comprising a composition comprising a benzodiazepine derivative or structural and functional related compound described in the present invention.

  Experiments conducted during the development of the present invention regulate various cellular processes associated with many conditions, including but not limited to viral infections, hyperproliferative autoimmune diseases, chronic inflammatory conditions, and cancer. A series of compounds used in the past have been identified. In some embodiments, the compound was benzodiazepine. In some embodiments, the compound was a benzodiazepine. In some embodiments, compounds having similar three-dimensional structural similarities (eg, similar presentation of functional groups in space) to the benzodiazepines of the present invention have also been found to function in the methods of the present invention. Thus, the present invention provides a broad class of compounds with special structural and functional properties used in the methods of the present invention.

Definitions To facilitate understanding of the invention, a number of terms and phrases are defined below.

As used herein, the term “benzodiazepine” refers to a 7-membered non-aromatic heterocycle fused to a phenyl ring, where the 7-membered ring is a nitrogen atom as part of the heterocycle. Has atoms. In some aspects, the two nitrogen atoms are in the 1 and 4 positions or the 1 and 5 positions, as shown in the general structure below.

  The term “greater than benzene” refers to any chemical group containing 7 or more non-hydrogen atoms.

  As used herein, the term “homologous” refers to similar properties as a result of having the same number of valence electrons in the same arrangement and is composed of different atoms, not necessarily the same number of atoms. Indicates one of two or more substances that are not composed of.

  The term “chemical moiety” refers to any compound containing at least one carbon atom. Examples of chemical moieties include, but are not limited to, aromatic chemical moieties, sulfur-containing chemical moieties, nitrogen-containing chemical moieties, hydrophilic chemical moieties, and hydrophobic chemical moieties.

  As used herein, the term “aliphatic” refers to groups that include, but are not limited to, alkyl, alkenyl, alkynyl, cycloaliphatic.

As used herein, the term “aryl” refers to a single aromatic ring, such as a phenyl ring, or two or more aromatic rings (eg, bisphenyl, naphthalene, anthracene), or aromatic An aromatic ring and one or more non-aromatic rings are shown. The aryl ring may be substituted with a lower aliphatic group (eg, alkyl, alkenyl, alkynyl, or cycloaliphatic). Additionally, the aliphatic and aryl groups, further -NH _2, -NHCOCH _3, -OH, lower alkoxy (C ₁ -C _4), halo (-F, -Cl, -Br or -I,) including It can be substituted with one or more functional groups that are not so limited.

  As used herein, the term “substituted aliphatic” means that at least one of the aliphatic hydrogen atoms is, for example, halogen, amino, hydroxy, nitro, thio, ketone, aldehyde, ester, amide, lower aliphatic Indicates an alkane, alkene, alkyne, or cycloaliphatic moiety that is substituted by a group, substituted lower aliphatic, or ring (aryl, substituted aryl, cycloaliphatic, substituted cycloaliphatic, etc.). Examples of such include, but are not limited to, 1-chloroethyl and the like.

  As used herein, the term “substituted aryl” means that at least one of the hydrogen atoms on the ring carbon is, for example, halogen, amino, hydroxy, nitro, thio, ketone, aldehyde, ester, amide, lower An aromatic ring or a fused aromatic ring composed of at least one aromatic ring substituted by an aliphatic, substituted lower aliphatic, or ring (aryl, substituted aryl, cycloaliphatic, or substituted cycloaliphatic) The structure is shown. Examples of such include, but are not limited to, hydroxyphenyl and the like.

  As used herein, the term “cycloaliphatic” refers to an aliphatic structure that includes a fused ring structure. Examples of such include, but are not limited to, decalin and the like.

  As used herein, the term “substituted cycloaliphatic” means that at least one of the aliphatic hydrogen atoms is halogen, nitro, thio, amino, hydroxy, ketone, aldehyde, ester, amide, lower aliphatic, A cycloaliphatic structure substituted by a substituted lower aliphatic or ring (aryl, substituted aryl, cycloaliphatic, or substituted cycloaliphatic) is shown. Examples of such include, but are not limited to, 1-chlorodecalyl, bicyclo-heptane, octane, and nonane (eg, norbornyl).

As used herein, the term “heterocycle” refers to an aromatic or non-aromatic ring containing, for example, one or more heteroatoms. The heteroatoms can be the same or different from each other. Examples of heteroatoms include, but are not limited to nitrogen, oxygen and sulfur. Aromatic and non-aromatic heterocycles are well known in the art. Some non-limiting examples of aromatic heterocycles include pyridine, pyrimidine, indole, purine, quinoline and isoquinoline. Non-limiting examples of non-aromatic heterocyclic compounds include piperidine, piperazine, morpholine, pyrrolidine and pyrazolidine. Examples of oxygen-containing heterocycles include, but are not limited to, furan, oxirane, 2H-pyran, 4H-pyran, 2H-chromene, and benzofuran. Examples of sulfur-containing heterocycles include, but are not limited to, thiophene, benzothiophene, and parathiazine. Examples of nitrogen-containing heterocycles include pyrrole, pyrrolodiline, pyrazole, pyrazolidine, imidazole, imidazoline, imidazolidine, pyridine, piperidine, prazine, piperazine, pyrimidine, indole, purine, benzimidazole, quinoline, isoquinoline, triazole, and triazine. But not limited to. Examples of heterocycles containing two different heteroatoms include, but are not limited to, phenothiazine, morpholine, parathiazine, oxazine, oxazole, thiazine, and thiazole. The heterocycle may be further substituted with one or more groups selected from aliphatic, nitro, acetyl (ie, —C (═O) —CH ₃ ), or aryl groups.

  As used herein, the term “substituted heterocycle” means that at least one of the ring carbon atoms is replaced by oxygen, nitrogen or sulfur and at least one of the aliphatic hydrogen atoms is halogen, hydroxy, thio, nitro. , Amino, ketone, aldehyde, ester, amide, lower aliphatic, substituted lower aliphatic, or heterocyclic structure substituted by a ring (aryl, substituted aryl, cycloaliphatic, or substituted cycloaliphatic). Examples of such include, but are not limited to, 2-chloropyranyl.

  As used herein, the term “linker” refers to a chain containing up to eight adjacent atoms that connect two different structural moieties, where such atoms are, for example, carbon Nitrogen, oxygen, or sulfur. Ethylene glycol is one non-limiting example.

  As used herein, the term “lower alkyl-substituted amino” refers to any alkyl unit containing up to 8 carbon atoms, wherein one of the aliphatic hydrogen atoms is replaced by an amino group. . Examples of such include, but are not limited to, ethylamino and the like.

  As used herein, the term “lower alkyl substituted halogen” refers to any alkyl unit containing up to 8 carbon atoms, wherein one of the aliphatic hydrogen atoms is replaced by a halogen. Examples of such include, but are not limited to, chloroethyl and the like.

  As used herein, the term “acetylamino” means any primary or secondary amino that is acetylated. Examples of such include, but are not limited to, acetamide and the like.

As used herein, the term “moiety involved in hydrogen bonding” or “chemical moiety involved in hydrogen bonding” refers to a group that accepts or donates a proton, thereby forming a hydrogen bond. . Some specific non-limiting examples of moieties involved in hydrogen bonding include fluoro, oxygen-containing and nitrogen-containing groups that are well known in the art. Some examples of oxygen-containing groups involved in hydrogen bonding include the following: hydroxy, lower alkoxy, lower carbonyl, lower carboxyl, lower ether and phenol groups. As used herein, the modifier “lower” refers to lower aliphatic groups (C ₁ -C ₄ ) to which individual oxygen-containing functional groups are attached. Thus, for example, the term “lower carbonyl” refers to formaldehyde, acetaldehyde, among others. Some non-limiting examples of nitrogen-containing groups involved in hydrogen bond formation include amino and amide groups. In addition, groups containing both oxygen and nitrogen atoms can also participate in hydrogen bond formation. Examples of such groups include nitro, N-hydroxy and nitrite groups. The hydrogen bond acceptor in the present invention may be a π electron of an aromatic ring.

  As used herein, the term “derivative” of a compound refers to a chemically modified compound, where the chemical modification is a functional group of the compound (eg, an aromatic ring) or a benzodiazepine backbone. Happens either. Such derivatives include esters of alcohol-containing compounds, esters of carboxy-containing compounds, amides of amino-containing compounds, amides of carboxy-containing compounds, imines of amino-containing compounds, acetals of aldehyde-containing compounds, ketals of carbonyl-containing compounds, and the like. But not limited to.

  As used herein, the term “subject” refers to an organism to be treated by the methods of the present invention. Such organisms include, but are not limited to, mammals (eg, mice, monkeys, horses, cows, pigs, dogs, cats, etc.) and most preferably include humans. In the context of the present invention, the term “subject” will generally refer to a treatment (eg, administration of a compound of the present invention and optionally one or more agents) for a condition characterized by dysregulation of the apoptotic process. Indicates an individual who has received or has already received.

  As used herein, the term `` diagnosed '' refers to signs and symptoms (e.g., resistance to conventional therapy), or disease due to genetic analysis, pathological analysis, histological analysis, etc. Indicates recognition.

  As used herein, the term “anticancer agent” or “conventional anticancer agent” refers to any chemotherapeutic compound, radiation therapy, or surgical procedure used in the treatment of cancer.

  As used herein, the term “in vitro” refers to an artificial environment and processes or reactions that occur within the artificial environment. In vitro environments include, but are not limited to, test tubes and cell cultures. The term “in vivo” refers to the natural environment (eg, animal or cell) and the processes or reactions that occur within the natural environment.

  As used herein, the term “host cell” refers to any eukaryotic or prokaryotic cell (eg, mammalian cell, avian cell, amphibian cell, plant cell), whether in vitro or in vivo. , Fish cells, and insect cells).

  As used herein, the term “cell culture” refers to any in vitro cell culture. The term is maintained in vitro, including continuous cell lines (e.g., having an immortal phenotype), primary cell cultures, finite cell lines (e.g., non-transformed cells), and oocytes and embryos. Any other group of cells is included.

  In some embodiments, “target cells” of the compositions and methods of the invention include, but are not limited to, lymphocyte cells or cancer cells. Lymphocytes include B cells, T cells, and granulocytes. Granulocytes include eosinophils and macrophages. In some embodiments, the target cell is a continuously cultured or uncultured cell obtained from a patient biopsy.

  Cancer cells include tumor cells, neoplastic cells, malignant cells, metastatic cells, and hyperplastic cells. Neoplastic cells can be benign or malignant. Neoplastic cells are benign if they do not invade and do not metastasize. Malignant cells are those that can invade and / or metastasize. Hyperplasia is a pathological accumulation in a tissue or organ and there is no significant change in structure or function.

  In one specific embodiment, the target cell exhibits pathological growth or proliferation. As used herein, the term “pathologically proliferating or growing cells” refers to a localized set of proliferating cells in an animal that is not governed by the normal limitations of normal growth.

As used herein, the term “non-activated target cell” refers to either a cell in the G ₀ phase or a cell to which no stimulus has been applied.

As used herein, the term “activated target lymphocyte cell” refers to a lymphocyte cell stimulated by an appropriate stimulus that causes a signaling cascade, or a lymphocyte cell that is not in G ₀ phase. Activated lymphocyte cells proliferate, undergo activation-induced cell death, or other characteristic of one or more cytotoxins, cytokines, and cell types (e.g., CD8 ⁺ or CD4 ⁺ ) May produce related membrane bound proteins. They can also recognize and bind to any target cell that displays a particular antigen on the surface and then release the effector molecule.

As used herein, the term “activated cancer cells” refers to cancer cells that have been stimulated with an appropriate stimulus that causes signal transduction. Activated cancer cells may or may not be in G ₀ phase.

An activator is a stimulus that, when interacting with a target cell, causes a signaling cascade. Examples of activating stimuli include, but are not limited to, small molecules, radiant energy, and molecules that bind to cell activated cell surface receptors. The response elicited by activating stimuli can be characterized, inter alia, by intracellular Ca ²⁺ , superoxide, or hydroxyl radical levels; the activity of enzymes such as kinases or phosphatases; or changes in cellular energy states. In cancer cells, the activator also contains a transforming oncogene.

  Examples of T cell ligands include, but are not limited to, peptides that bind to MHC molecules, peptide MHC complexes, or antibodies that recognize T cell receptor components.

  Examples of B cell ligands include, but are not limited to, molecules or antibodies that bind to or recognize a component of the B cell receptor.

  Examples of mediators or conditions that enhance cellular stress include, but are not limited to, heat, radiation, oxidative stress, or growth factor withdrawal. Examples of growth factors include, but are not limited to, serum, IL-2, platelet derived growth factor (“PDGF”), and the like.

  As used herein, the term “effective amount” refers to the amount of a compound (eg, a compound of the invention) sufficient to achieve a beneficial or desired result. An effective amount can be administered in one or more administrations, applications or doses, and is not intended to be limiting when limited to a particular formulation or route of administration.

  As used herein, the term “abnormal regulation of the process of cell death” refers to any abnormality in the ability (eg, predisposition) of a cell to undergo cell death by either necrosis or apoptosis. Abnormal regulation of cell death is associated with or induced by various conditions, such as autoimmune diseases (systemic lupus erythematosus, rheumatoid arthritis, graft-versus-host disease, myasthenia gravis, Sjogren's syndrome, etc.) Chronic inflammation conditions (e.g. psoriasis, asthma and Crohn's disease), hyperproliferative diseases (e.g. tumors, B cell lymphomas, T cell lymphomas, etc.), viral infections (e.g. herpes, papilloma, HIV), and osteoarthritis and Other conditions include atherosclerosis.

  It should be noted that if the dysregulation is induced by or associated with a viral infection, the viral infection may or may not be detected when the dysregulation occurs or is observed. is there. That is, virus-induced dysregulation can occur even after the symptoms of viral infection have disappeared.

  As used herein, “hyperproliferative disease” refers to any condition in which the localized population of proliferating cells in an animal is not limited by the normal limitations of normal growth. Examples of hyperproliferative diseases include tumors, neoplasms, lymphomas and the like. Neoplasms are said to be benign if they cannot invade or metastasize, or malignant if either. A metastatic cell or tissue means that the cell can invade and destroy adjacent body structures. Hyperplasia is a form of cell proliferation in which there is no significant change in structure or function involving an increase in the number of cells in a tissue or organ. Dysplasia is a form of controlled cell growth in which one type of fully differentiated cell replaces another type of differentiated cell. Dysplasia can occur in epithelial or connective tissue cells. Typical dysplasia involves a somewhat disordered dysplastic epithelium.

  Pathological proliferation of activated lymphocyte cells often causes an autoimmune disease or a chronic inflammatory condition. As used herein, “autoimmune disease” refers to any condition in which an organism produces antibodies or immune cells that recognize the organism's own molecules, cells or tissues. Non-limiting examples of autoimmune diseases include autoimmune hemolytic anemia, autoimmune hepatitis, Berger's disease or IgA nephropathy, Celiac sprue, chronic fatigue syndrome, Crohn's disease, dermatomyositis, fibromyalgia, graft Anti-host disease, Graves' disease, Hashimoto's thyroiditis, idiopathic thrombocythemia, lichen planus, multiple sclerosis, myasthenia gravis, psoriasis, rheumatic fever, rheumatoid arthritis, scleroderma, Sjogren's syndrome, systemic lupus erythematosus , Type I diabetes, ulcerative colitis, vitiligo, tuberculosis and the like.

  As used herein, the term “chronic inflammatory condition” refers to a condition in which an organism's immune cells are activated. Such a condition is characterized by a persistent inflammatory response with pathological sequelae. This condition is characterized by mononuclear cell infiltration, fibroblast and small vessel proliferation, increased connective tissue, and tissue destruction. Examples of chronic inflammatory diseases include, but are not limited to, Crohn's disease, chronic obstructive pulmonary disease, inflammatory bowel disease, multiple sclerosis, and asthma. Autoimmune diseases such as rheumatoid arthritis and systemic lupus erythematosus can also cause chronic inflammatory conditions.

  As used herein, the term “simultaneous administration” refers to the administration of at least two agents (eg, a compound of the invention) or therapy to a subject. In some embodiments, co-administration of two or more drugs / therapies is performed simultaneously. In other embodiments, the first agent / therapy is administered before the second agent / therapy. One skilled in the art will appreciate that the various drug / therapeutic formulations used and / or the route of administration may vary. Appropriate doses for simultaneous administration can be readily determined by one skilled in the art. In some embodiments, when drugs / therapies are co-administered, the individual drugs / therapies are administered at a dose that is lower than the dose suitable for their single administration. Thus, co-administration is particularly desirable in embodiments where co-administration of drugs / therapies reduces the required dose of known potentially harmful (eg, toxic) drugs.

  As used herein, the term “toxicity” refers to any adverse or deleterious effect on the same cell or tissue as compared to the cell or tissue prior to administration of the toxicant.

  As used herein, the term “pharmaceutical composition” refers to an inactive or active carrier and active agent, making them particularly suitable for in vivo, in vivo or ex vivo diagnostic or therapeutic applications. The combination of is shown.

  As used herein, the term “pharmaceutically acceptable carrier” refers to phosphate buffered saline, water, emulsions such as oil / water or water / oil emulsions, and various Indicates any standard pharmaceutical carrier, such as a type of wetting agent. The composition can also include stabilizers and preservatives. Examples of carriers are stabilizers and adjuvants (see, eg, Martin, Remington's Pharmaceutical Sciences, 15th Ed., Mack Publ. Co., Easton, PA [1975]).

  As used herein, the term “pharmaceutically acceptable salt” refers to a compound of the invention, or an active metabolite or residue thereof, that can provide a compound of the invention when administered to a subject. Any pharmaceutically acceptable salt (eg acid or base) of the compound As is known to those skilled in the art, “salts” of the compounds of the present invention may be derived from inorganic or organic acids and bases. Examples of acids include hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, perchloric acid, fumaric acid, maleic acid, phosphoric acid, glycolic acid, lactic acid, salicylic acid, succinic acid, toluene-p-sulfonic acid, tartaric acid, acetic acid , Citric acid, methanesulfonic acid, ethanesulfonic acid, formic acid, benzoic acid, malonic acid, naphthalene-2-sulfonic acid, benzenesulfonic acid, and the like. Other acids such as oxalic acid are not pharmaceutically acceptable per se, but are used in the preparation of salts useful as intermediates in obtaining the compounds of the invention and their pharmaceutically acceptable acid addition salts. May be.

Examples of bases include, but are not limited to, alkali metal (eg, sodium) hydroxide, alkaline earth metal (eg, magnesium) hydroxide, ammonia, and compounds of formula NW ₄ ^+. Where W is C _1-4 alkyl and the like.

Examples of salts include, but are not limited to: acetate, adipate, alginate, aspartate, benzoate, benzenesulfonate, bisulphate, butyrate, citrate, camphorate, camphorsulfonate, cyclopentanepropionate, digluconate , Dodecyl sulfate, ethane sulfonate, fumarate, flucoheptanoate, glycerophosphate, hemisulphate, heptanoate, hexanoate, hydrochloride, hydrobromide, hydroiodide, 2-hydroxyethane sulfonate, lactate, maleate, methane sulfonate, 2-naphthalene sulfonate, Nicotinate, oxalate, palmoate, pectinate, persulfate, phenylpropionate, Picrate, pivalate, propionate, succinate, tartrate, thiocyanate, tosylate, undecanoart, etc. Other examples of salts include anions of compounds of the present invention combined with a suitable cation such as Na ⁺ , NH ₄ ⁺ , and NW ₄ ⁺ , where W is a C _1-4 alkyl group Is mentioned.

  For therapeutic use, the compounds of the invention are intended to be pharmaceutically acceptable. However, pharmaceutically unacceptable acid and base salts may also be used in the preparation or purification of pharmaceutically acceptable compounds.

  As used herein, the term “pathogen” refers to a biological agent that causes a disease state (eg, infection, cancer, etc.) in a host. “Pathogens” include, but are not limited to, viruses, bacteria, archaea, fungi, protozoa, mycoplasma, prions, and parasites.

  “Bacteria” and “bacteria” refer to all prokaryotes, including those within all gates of the prokaryotic kingdom. The term is intended to include all microorganisms considered to be bacteria including mycoplasma, chlamydia, actinomyces, streptomyces and rickettsia. All forms of bacteria are included in this definition, including cocci, bacilli, spirochetes, spheroplasts, protoplasts, and the like. Included within this term are prokaryotes that are Gram negative or Gram positive. “Gram negative” and “Gram positive” indicate staining patterns by the Gram staining process well known in the art (eg, Finegold and Martin, Diagnostic Microbiology, 6th Ed., CV Mosby St. Louis, pp. 13-15 [ 1982]). “Gram-positive bacteria” are bacteria that retain the primary dye used in Gram staining, and the stained cells show a dark blue to purple color under the microscope. “Gram-negative bacteria” do not retain the primary dye used in Gram staining, but are stained by counterstaining. Thus, Gram negative bacteria show a red color.

  As used herein, the term “microorganism” refers to any species or type of microorganism, including but not limited to bacteria, archaea, fungi, protozoa, mycoplasma, and parasites. The present invention considers that many microorganisms included in the present invention are also pathogens for a subject.

  As used herein, the term “fungus” is used in reference to eukaryotes such as molds and yeasts, including dimorphic fungi.

  As used herein, the term “virus” is a fine infection that, with certain exceptions, cannot be observed with a light microscope, lacks independent metabolism, and can only replicate in living host cells. Indicates pathogen. Individual particles (ie, virions) are typically composed of a nucleic acid and a protein shell or coat; some virions also have a lipid-containing membrane. The term “virus” includes all types of viruses, including animals, plants, phages, and other viruses.

  As used herein, the term “sample” is used in its broadest sense. Samples suspected of exhibiting a condition characterized by dysregulation of apoptotic function include cells, tissues, or fluids, chromosomes isolated from cells (e.g., metaphase chromosome spread), genomic DNA (in solution or Southern blot analysis) RNA (coupled to a solid support, such as in solution or for Northern blot analysis), cDNA (coupled to a solid support, such as for a northern support), etc. But you can. Samples suspected of containing proteins may include cells, portions of tissue, extracts containing one or more proteins, and the like.

  As used herein, the term “purified” or “purify” refers to the removal of unwanted components from a sample. As used herein, the term “substantially purified” means that there is at least 60%, preferably 75%, most preferably 90% or more of other components normally associated with it. Indicates a molecule.

  As used herein, the term “antigen binding protein” refers to a protein that binds to a specific antigen. “Antigen binding proteins” include, but are not limited to, polyclonal, monoclonal, chimeric, single chain, and humanized antibodies, Fab fragments, F (ab ′) 2 fragments, and immunoglobulins including Fab expression libraries. . Various procedures known in the art are used to produce polyclonal antibodies. For antibody production, various host animals can be immunized by injection of peptides corresponding to the desired epitope, including but not limited to rabbits, mice, rats, sheep, goats and the like. In preferred embodiments, the peptide is conjugated to an immunogenic carrier such as diphtheria toxoid, bovine serum albumin (BSA), or keyhole limpet hemocyanin [KLH]. Depending on the host species, various adjuvants are used to increase the immune response, including Freund (complete and incomplete), mineral gels such as sodium hydroxide, lysolecithin, pluronic polyols, polyanions, peptides, oil emulsions, Surface active substances such as keyhole limpet hemocyanin, dinitrophenol, and potentially useful human adjuvants such as BCG (Bacille Calmette Guerin) and Cholinebacterium parvum are included, but are not limited thereto.

  For preparation of monoclonal antibodies, any technique that produces antibody molecules in a continuous cell line in culture may be used (e.g. Harlow and Lane, Antibodies: A Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring See Harbor, NY [1975]). These include hybridoma technology originally developed by Kohler and Milstein (Kohler and Milstein, Nature, 256: 495-497), as well as trioma technology, human B-cell hybridoma technology (e.g., Kozbor et al., Immunol. Today, 4:72 [1983]), and EBV-hybridoma technology for producing human monoclonal antibodies (Cole et al., In Monoclonal Antibodies and Cancer Therapy, Alan R. Liss, Inc., pp. 77- 96 [1985]), but is not limited thereto.

  According to the present invention, techniques described for the production of single chain antibodies (U.S. 4,946,778; incorporated herein by reference) can be adapted to produce the desired specific single chain antibody. In another embodiment of the present invention, a technique known in the art for generating Fab expression libraries (Huse et al., Science, 246: 1275-1281 [1989]) is used to have the desired specificity. Allows rapid and easy identification of monoclonal Fab fragments.

  Antibody fragments containing the idiotype (antigen binding region) of the antibody molecule can be generated by known techniques. For example, such fragments include, but are not limited to: F (ab ′) 2 fragments that can be generated by pepsin digestion of antibody molecules; reducing disulfide bridges of F (ab ′) 2 fragments The Fab ′ fragment that can be generated by and the Fab fragment that can be generated by treating antibody molecules with papain and a reducing agent.

  The gene encoding the antigen binding protein can be isolated by a method known in the art. For antibody production, screening for the desired antibody involves techniques known in the art (e.g., radioimmunoassay, ELIZA (enzyme linked immunosorbent assay), "sandwich" immunoassay, immunoradiometric assay, gel diffusion precipitation reaction, immunoassay). Diffusion assays, in situ immunoassays (e.g., use of colloidal gold, enzyme or radioisotope labels), Western blots, precipitation reactions, agglutination assays (e.g., gel aggregation assays, hemagglutination assays, etc.), complement binding assays, immunizations Fluorescent assays, protein A assays, and immunoelectrophoretic assays, etc.) and the like.

As used herein, the term “immunoglobulin” or “antibody” refers to a protein that binds to a specific antigen. Immunoglobulins include, but are not limited to, polyclonal, monoclonal, chimeric, and humanized antibodies, Fab fragments, F (ab ′) ₂ fragments, including the following classes of immunoglobulins: IgG, IgA, IgM , IgD, IbE, and secreted immunoglobulin (sIg). An immunoglobulin generally comprises two identical heavy chains and two light chains. However, the terms “antibody” and “immunoglobulin” also include single chain antibodies and double chain antibodies.

  As used herein, the term “epitope” refers to the portion of an antigen that makes contact with a particular immunoglobulin. When immunizing a host animal using a protein or a fragment of a protein, many regions of the protein can induce the production of antibodies that specifically bind to certain regions or three-dimensional structures of the protein; these regions Or the structure is called an “antigenic determinant”. An antigenic determinant can compete with an intact antigen (ie, an “immunogen” used to elicit an immune response) for binding to an antibody.

  The terms `` specific binding '' or `` specific binding '' when used in connection with antibody and protein or peptide interactions are those in which the interaction is a special structure of the protein (i.e., antigenic determinant or epitope). Relying on existence; in other words, it means that an antibody recognizes and binds to a specific protein structure rather than a general protein. For example, if the antibody is specific for epitope “A”, in reactions involving label “A” and the antibody, the presence of a protein containing epitope A (or free, unlabeled A) indicates that the label bound to the antibody Reduce the amount of A.

  As used herein, the terms “non-specific binding” and “background binding”, when used in reference to an antibody and protein or peptide interaction, refer to an interaction that is independent of the presence of a particular structure. (Ie, the antibody binds to a general protein rather than a special structure such as an epitope).

  As used herein, the term `` modulate '' affects (e.g., promotes or interferes with) aspects of cell function, including but not limited to cell growth, proliferation, apoptosis, etc. Shows the activity of a compound (eg, a compound of the invention).

As used herein, the term "competing for binding" refers to a second molecule (e.g., a second compound of the invention or other that binds to an oligomycin sensitizing protein in mitochondrial ATP synthase). Is used for the first molecule (eg, the first compound of the present invention) having the activity of binding to the same substance (eg, the oligomycin sensitizing protein in mitochondrial ATP synthase). The efficiency of binding by the first molecule (eg, kinetics or thermodynamics) may be the same, greater than, or less than the efficiency of the substance that binds to the second molecule. For example, the substance equilibrium binding constant (K _D ) may be different for the two molecules. As used herein, the term “instruction for administering a compound to a subject” and its grammatical equivalents are for the treatment of conditions characterized by abnormal regulation of the apoptotic process in a cell or tissue. Instructions for using the composition included in the kit (eg, providing the dose, route of administration, decision tree for the treating physician to correlate patient-specific characteristics with the course of action). The term also includes autoimmune diseases (e.g. systemic lupus erythematosus, rheumatoid arthritis, graft-versus-host disease, myasthenia gravis, Sjogren's syndrome, etc.), chronic inflammatory diseases (e.g., psoriasis, asthma and Crohn's disease), Treat hyperproliferative diseases (e.g., tumors, B-cell lymphoma, T-cell lymphoma, etc.), viral infections (e.g., herpes virus, papilloma virus, HIV), and other conditions such as osteoarthritis and atherosclerosis Specific instructions for the use of the composition contained in the kit are provided.

  The term “test compound” is used to treat or prevent a disease, disorder, disease, or disorder of bodily function, or otherwise, the physiological or cellular state of a sample (eg, an abnormality of apoptosis in a cell or tissue). Indicates any chemical entity, drug, drug, etc. that can be used to change the level of regulation). Test compounds include known and potential therapeutic compounds. A test compound can be determined to be therapeutic using the screening methods of the invention. “Known therapeutic compounds” refers to therapeutic compounds that have been shown to be effective in such treatment or prevention (eg, through animal studies or prior to administration to humans). In some embodiments, a “test compound” is an agent that modulates apoptosis in a cell.

General Description of the Invention As a class of drugs, benzodiazepine compounds have been extensively studied and reported to be effective agents for treating many diseases. For example, US 4,076823, US 4,110,337, US 4,495,101, US 4,751,223, and US 5,776,946, each of which is incorporated herein by reference in its entirety, are benzodiazepine compounds that are effective as analgesics and anti-inflammatory agents. It is reported that there is. Similarly, US 5,324,726 and US 5,597,915, each incorporated herein by reference in their entirety, are benzodiazepine compounds that are antagonists of cholecystokinin and gastrin, and thus treat certain gastrointestinal diseases. It is reported that it is useful.

  Other benzodiazepine compounds have been studied as inhibitors of human neutrophil elastase, particularly in the treatment of human neutrophil elastase-mediated conditions such as myocardial ischemia, septic shock syndrome (see, for example, the present specification in its entirety. (See US 5,861,380 incorporated by reference). U.S. 5,041,438, incorporated herein by reference in its entirety, reports usefulness as an antiretroviral agent.

  Despite the interest of benzodiazepine compounds, it is noted that the present invention provides novel benzodiazepine compounds and related compounds as well as novel compounds and methods of using known compounds for the treatment of various diseases. It will be clear from the explanation.

  Benzodiazepine compounds are known to bind to benzodiazepine receptors within the central nervous system (CNS) and have therefore been used to treat a variety of CNS disorders including anxiety and epilepsy. A peripheral benzodiazepine receptor has also been identified, and this receptor can also be incidentally present in the CNS. The present invention demonstrates that benzodiazepines and related compounds have pro-apoptotic and cytotoxic properties useful in the treatment of transformed cells grown in tissue culture. The pathway of action of these compounds is not mediated by the previously identified benzodiazepine receptors.

  Thus, in some aspects, the present invention provides a number of new compounds and previously known compounds that are induced against new cellular targets to achieve the desired biological result. In another aspect, the present invention provides a method for using such compounds to modulate biological processes. The present invention also provides drug screening methods for identifying and optimizing compounds. The invention further provides diagnostic markers for identifying diseases and conditions, monitoring treatment plans, and / or identifying optimal treatment strategies. These and other research and therapeutic utilities are described below.

Detailed Description of the Invention The present invention provides novel compounds, methods for their discovery, their therapeutic, research and diagnostic applications. In particular, the present invention treats benzodiazepine derivatives and related compounds, and benzodiazepine derivatives and related compounds, for many conditions associated with dysregulation of processes such as programmed cell death, autoimmunity, inflammation, and hyperproliferation. A method for use as a therapeutic agent is provided.

  Exemplary compositions and methods of the present invention are described in greater detail in the following sections: I. Modulators of cell death; II. Exemplary compounds; III. Pharmaceutical compositions, formulations, and exemplary administration Routes and administration considerations; IV. Drug screening; and V. Treatment applications.

  Unless stated otherwise, the practice of the present invention includes organic chemistry, pharmacology, molecular biology (including recombinant technology), cell biology, biochemistry, and immunology within the skill of the art. Use conventional techniques. Such techniques are described, for example, in “Molecular cloning: a laboratory manual” Second Edition (Sambrook et al., 1989); “Oligonucleotide synthesis” (MJ Gait, ed., Each incorporated herein by reference in its entirety. "Animal cell culture" (RI Freshney, ed., 1987); Series "Methods in enzymology" (Academic Press, Inc.); "Handbook of experimental immunology" (DM Weir & CC Blackwell, eds.); “Gene transfer vectors for mammalian cells” (JM Miller & MP Calos, eds., 1987); “Current protocols in molecular biology” (FM Ausubel et al., Eds., 1987, and periodic updates); “PCR: The polymerase chain reaction ”(Mullis et al., eds., 1994); and“ Current protocols in immunology ”(JE Coligan et al., eds., 1991) are fully explained.

I. Modulators of cell death In some embodiments, the present invention modulates apoptosis by exposing cells to a compound. The effect of a compound can be measured by detecting a significant number of cellular changes. Cell death may be assayed as described herein and in the art. In some embodiments, the cell line appropriate cell culture conditions (e.g., gas (CO _2), temperature and media) below, during the appropriate period of density dependence unrestricted exponential growth is achieved, maintained Is done. Cell number and / or viability is measured using standard techniques such as trypan blue exclusion / cytometry, or MTT dye conversion assay. The cells may also be analyzed for expression of genes or gene products associated with abnormalities of apoptosis or necrosis.

In some embodiments, apoptosis is induced when the present invention is exposed to cells. In some embodiments, the present invention induces apoptosis through interaction with mitochondrial F ₁ F ₀ -ATPase.

II. Exemplary Compounds Exemplary compounds of the invention are shown below. Certain embodiments of the present invention include benzodiazepine compounds having a chemical moiety that causes the benzodiazepine to lack an asymmetric center associated with the third carbon position of the benzodiazepine ring.

RおよびS鏡像体の両方ならびにラセミ混合物が含まれる。 Certain embodiments of the present invention include compositions comprising a benzodiazepine compound having a chemical moiety that causes the benzodiazepine to lack an asymmetric center associated with the third carbon position of the benzodiazepine ring. In some embodiments, the composition comprises the following formula:

Both R and S enantiomers as well as racemic mixtures are included.

In some embodiments, A ---- B is selected from the group consisting of N-CH ₂ and C = N.

  In some embodiments, R5 is a linker group and is present or absent.

  In some embodiments, R1 is an isostere of OH. In some embodiments, R1 is hydrogen; halogen; OH; a chemical moiety comprising an aryl subgroup; a chemical moiety comprising a substituted aryl subgroup; a chemical moiety comprising a cycloaliphatic subgroup; a chemical moiety comprising a substituted cycloaliphatic subgroup A chemical moiety comprising a heterocyclic subgroup; a chemical moiety comprising a substituted heterocyclic subgroup; a chemical moiety comprising at least one ester subgroup; a chemical moiety comprising at least one ether subgroup; a straight chain or branched, saturated or An aliphatic chain having at least two carbons, unsaturated, substituted or unsubstituted; a chemical moiety containing sulfur; a chemical moiety containing nitrogen; and -OR-, wherein R is an aryl subgroup Chemical moieties including substituted aryl subgroups; chemical moieties including cycloaliphatic subgroups; chemical moieties including substituted cycloaliphatic subgroups; chemical moieties including heterocyclic subgroups; A chemical moiety comprising a ring subgroup; a linear or branched, saturated or unsaturated, substituted or unsubstituted aliphatic chain having at least two carbons; a chemical moiety comprising at least one ester subgroup; at least one ether sub Selected from the group consisting of a chemical moiety comprising a group; a chemical moiety comprising sulfur; and a chemical moiety comprising nitrogen.

式中、R1’は、ハロゲン；アルキル；置換アルキル；アリール；置換アリール；アミノ；カルボニル；スルホン；スルホンアミド；エーテル；OH；硫黄を含む化学部分；窒素を含む化学部分；CH₃；直鎖または分枝、飽和または不飽和の少なくとも1つの炭素を有する脂肪族鎖；直鎖または分枝、飽和または不飽和の少なくとも2つの炭素を有し、少なくとも1つのヒドロキシサブ基を有する脂肪族鎖；直鎖または分枝、飽和または不飽和の少なくとも2つの炭素を有し、少なくとも1つのチオールサブ基を有する脂肪族鎖；直鎖または分枝、飽和または不飽和の少なくとも2つの炭素を有し、アルデヒドサブ基で終わる脂肪族鎖；直鎖または分枝、飽和または不飽和の少なくとも2つの炭素を有し、少なくとも1つのケトンサブ基を有する脂肪族鎖；直鎖または分枝、飽和または不飽和の少なくとも2つの炭素を有し、カルボン酸サブ基で終わる脂肪族鎖；直鎖または分枝、飽和または不飽和の少なくとも2つの炭素を有し、少なくとも1つのアミドサブ基を有する脂肪族鎖；直鎖または分枝、飽和または不飽和の少なくとも2つの炭素を有し、少なくとも1つのアシル基を有する脂肪族鎖；直鎖または分枝、飽和または不飽和の少なくとも2つの炭素を有し、少なくとも1つの窒素含有部分を有する脂肪族鎖；直鎖または分枝、飽和または不飽和の少なくとも2つの炭素を有し、少なくとも1つのアミンサブ基を有する脂肪族鎖；直鎖または分枝、飽和または不飽和の少なくとも2つの炭素を有し、少なくとも1つのエーテルサブ基を有する脂肪族鎖；直鎖または分枝、飽和または不飽和の少なくとも2つの炭素を有し、少なくとも1つのハロゲンサブ基を有する脂肪族鎖；直鎖または分枝、飽和または不飽和の少なくとも2つの炭素を有し、少なくとも1つのニトロニウムサブ基を有する脂肪族鎖からなる群より選択される。 In some embodiments, R1 is selected from the group consisting of:

Wherein, R1 'is halogen, alkyl, substituted alkyl, aryl, substituted aryl; amino; carbonyl; sulfone; sulfonamide; ethers; OH; chemical moieties which contain a nitrogen; chemical moiety containing a sulfur CH _3; straight or An aliphatic chain having at least one carbon that is branched, saturated or unsaturated; an aliphatic chain having at least two carbons that is linear or branched, saturated or unsaturated, and having at least one hydroxy subgroup; An aliphatic chain having at least two carbons that are chain or branched, saturated or unsaturated, and having at least one thiol subgroup; having at least two carbons that are linear or branched, saturated or unsaturated, and having an aldehyde sub Aliphatic chain ending in a group; straight chain or branched, saturated or unsaturated, aliphatic chain having at least one ketone subgroup; straight chain Or an aliphatic chain having at least two branched, saturated or unsaturated carbons and ending with a carboxylic acid subgroup; having at least two carbons that are straight or branched, saturated or unsaturated, and at least one An aliphatic chain having an amide subgroup; an aliphatic chain having at least two carbons that are linear or branched, saturated or unsaturated, and having at least one acyl group; at least one of linear or branched, saturated or unsaturated An aliphatic chain having two carbons and having at least one nitrogen-containing moiety; an aliphatic chain having at least two carbons that are linear or branched, saturated or unsaturated, and having at least one amine subgroup; An aliphatic chain having at least two carbons that are chain or branched, saturated or unsaturated, and having at least one ether subgroup; at least two carbons that are linear or branched, saturated or unsaturated An aliphatic chain having at least one halogen subgroup; a group consisting of an aliphatic chain having at least two carbons, straight or branched, saturated or unsaturated, and having at least one nitronium subgroup More selected.

  In some embodiments, R2 is a chemical moiety comprising an aryl subgroup; a chemical moiety comprising a substituted aryl subgroup; a chemical moiety comprising a cycloaliphatic subgroup; a chemical moiety comprising a substituted cycloaliphatic subgroup; a heterocyclic subgroup A chemical moiety comprising a substituted heterocyclic subgroup; a chemical moiety comprising sulfur; and a chemical moiety comprising nitrogen. In some embodiments, R2 is a cyclic group larger than benzene, where greater than benzene includes any chemical group containing 7 or more non-hydrogen atoms.

；キノリン、および全ての芳香族位置異性体からなる群より選択される。 In some embodiments, R2 is naphthalene; phenol; 1-naphthalenol; 2-naphthalenol;

Selected from the group consisting of quinoline and all aromatic positional isomers.

  In some embodiments, R3 is an isostere of OH. In some embodiments, R3 is hydrogen; halogen; OH; a chemical moiety comprising at least one ester subgroup; a chemical moiety comprising at least one ether subgroup; linear or branched, saturated or unsaturated, substituted or An aliphatic chain having at least two unsubstituted carbons; a chemical moiety containing sulfur; a chemical moiety containing nitrogen; and -OR-, wherein R is a chemical moiety containing an aryl subgroup; A chemical moiety comprising an aryl subgroup; a chemical moiety comprising a cycloaliphatic subgroup; a chemical moiety comprising a substituted cycloaliphatic subgroup; a chemical moiety comprising a heterocyclic subgroup; a chemical moiety comprising a substituted heterocyclic subgroup; Branched, saturated or unsaturated, substituted or unsubstituted aliphatic chain having at least two carbons; at least one ester sub-group containing a chemical moiety; at least one ether sub-group containing It is selected from the group consisting of chemical moiety comprising nitrogen; chemical moiety comprising sulfur; chemical moiety.

In some embodiments, R3 is alkyl; mono-substituted alkyl; disubstituted alkyl; trisubstituted alkyl; (CH _{2) n,} wherein _{n = 1~6; CN; N 3} ; CNO; NH 2; SH; CF _{3; OCH 3; NCH 2 CH} (CH 2) N (CH 3) 2; NCH 2 CHCH 2 N (CH 3) 2; phenyl; 2-pyridyl; 3-pyridyl; 4-pyridyl; NCH _3; NCONHCH _{3; CH 2 OH; NHCONH 2; NHCOCH} 3; NHSO 2 CH 3; NHCN; NHCHO; SOCH 3; SO 2 CH 3; CHNOH; CHNOCH 3; SCH 3; CH 2 CO; CH 2 SO 2; CONH; CH 2 C ( _{NOH); CH 2 C (nOMe} ); NHSO 2 PH; NHCS; CH 2 NHCO; is selected from the group consisting of and NHCOS; COCH _2; NHCO _2. In some embodiments, R3 is an isotope described, for example, in Patani, G. and LaVoie, EJ, 1996, Chem. Rev. 96: 3147-3176, which is incorporated herein by reference in its entirety. Either is explained.

  In some embodiments, R3 is a cyclic structure attached at the 6,7 carbon position of the benzodiazepine structure, the 7,8 carbon position of the benzodiazepine structure, or the 8,9 carbon position of the benzodiazepine structure. In some embodiments, the cyclic structure comprises a chemical moiety comprising an aryl subgroup; a chemical moiety comprising a substituted aryl subgroup; a chemical moiety comprising a cycloaliphatic subgroup; a chemical moiety comprising a substituted cycloaliphatic subgroup; a heterocyclic subgroup A chemical moiety containing a substituted heterocyclic subgroup; a chemical moiety containing sulfur; a chemical moiety containing oxygen; and a chemical moiety containing nitrogen.

  In some embodiments, at least one of R1 and R3 is a chemical moiety that participates in hydrogen bonding. In some embodiments, the distance between the chemical moiety involved in hydrogen bonding and the R2 group in three-dimensional space differs, for example, by only about 12 angstroms.

であり、ここで、R4’は直鎖または分枝、飽和または不飽和、置換または非置換の少なくとも2つの炭素を有する脂肪族鎖である。 In some embodiments, R4 is a chemical moiety that causes benzodiazepine to lack an asymmetric center. In some embodiments, R4 is hydrogen,

Where R 4 ′ is a linear or branched, saturated or unsaturated, substituted or unsubstituted aliphatic chain having at least two carbons.

In some embodiments, the composition comprises the following chemical formula:

In some embodiments, the composition comprises a formula selected from the group consisting of:

であり；式中、R4''は1〜6個の炭素であり、そのいずれか1つは置換されるか、または非置換である。置換炭素としては、OH；ハロゲン；CH₃；直鎖または分枝、環状または非環状、飽和または不飽和の少なくとも2つの炭素を有する脂肪族鎖；ハロゲンを含む化学部分；硫黄を含む化学部分；窒素を含む化学部分；芳香族化学部分；親水性化学部分；および疎水性化学部分で置換されたものが挙げられるが、それらに限定されない。 In some embodiments, the structure is

Wherein R 4 ″ is 1 to 6 carbons, any one of which is substituted or unsubstituted. Substituted carbons include: OH; halogen; CH ₃ ; linear or branched, cyclic or acyclic, saturated or unsaturated aliphatic chain having at least two carbons; halogen containing chemical moiety; sulfur containing chemical moiety; Chemical moieties including nitrogen; aromatic chemical moieties; hydrophilic chemical moieties; and those substituted with hydrophobic chemical moieties include, but are not limited to.

In some embodiments, the compound is selected from the group consisting of:

In some embodiments, the compound is selected from the group consisting of:

  From the above description, it is clear that many specific examples are represented by the above general formula. Various sub-combinations resulting from the selection of a particular group at each substituent position are possible and all such sub-combinations are within the scope of the present invention.

  Further, it is to be understood that the numerical ranges given throughout this disclosure should be considered as flexible ranges intended for any possible subregion within that range. For example, descriptions of groups having a range of 1-10 carbons are also, for example, 1-3, 1-5, 1-8, or 2-3, 2-5, 2-8 , 3-4, 3-5, 3-7, 3-9, 3-10, etc., groups having carbon subregions are contemplated. Thus, the range of 1-10 should be considered as the outer boundary of the range where many possible subregions are clearly intended. Other examples that are intended to be ranges in other contexts are found throughout this disclosure, where such ranges include similar subregions therein.

In summary, a number of compounds are provided herein. Any one or more of these compounds can be used to treat a variety of dysregulated diseases associated with cell death as described elsewhere herein. Furthermore, any one or more of these compounds can be used to block ATP hydrolysis without affecting cell synthesis or cell viability. Further, any one or more of these compounds may be combined with at least one other therapeutic agent (e.g., potassium channel opener, calcium channel blocker, sodium hydrogen exchange inhibitor, antiarrhythmic agent, antiatherosclerosis agent, Coagulant, antithrombotic, prothrombolytic, fibrinogen antagonist, diuretic, antihypertensive, ATPase inhibitor, mineralocorticoid receptor antagonist, phosphodiesterase inhibitor, antidiabetic, antiinflammatory, antioxidant, Angiogenesis modulators, anti-osteoporosis drugs, hormone replacement therapy, hormone receptor modulators, oral contraceptives, anti-obesity drugs, antidepressants, anxiolytics, antipsychotics, anti-proliferative drugs, anti-tumor drugs, anti-ulcers and gastroesophageal Reflux disease drug, growth hormone drug and / or growth hormone secretagogue, thyroid mimetic, anti-infective, antiviral, antibacterial, antifungal , Cholesterol / lipid lowering agents and lipid profile therapy, and drugs that mimic ischemic preconditioning and / or myocardial stunning, anti-atherosclerotic agents, anticoagulants, antithrombotics, antihypertensives, antidiabetics, and ACE inhibition Agents, AT-1 receptor antagonists, ET receptor antagonists, dual ET / AII receptor antagonists, and antihypertensive agents selected from vasopepsidase inhibitors, or GPIIb / IIIa blockers, P2Y ₁ and P2Y ₁₂ antagonists , Thromboxane receptor antagonists, and antiplatelet agents selected from aspirin) and can be used in pharmaceutical compositions with pharmaceutically acceptable carriers or diluents. Furthermore, using any one or more of these compounds, mitochondrial F ₁ F ₀ ATP hydrolase related diseases in patients (e.g. myocardial infarction, ventricular hypertrophy, coronary artery disease, non-Q wave myocardial infarction, congestive heart failure, Arrhythmia, Unstable angina, Chronic stable angina, Prinzmetal angina, Hypertension, Intermittent claudication, Obstructive peripheral arterial disease, Thrombus or thrombus obstruction symptoms of thrombo-occlusive stroke, Venous thrombosis, Arterial thrombosis , Cerebral thrombus, pulmonary thrombosis, cerebral embolism, thrombus formation tendency, disseminated intravascular coagulation, restenosis, atrial fibrillation, ventricular dilatation, atherosclerotic vascular disease, atherosclerotic plaque failure, atherosclerotic plaque formation, transplantation atheroma Atherosclerosis, vascular remodeling atherosclerosis, cancer, surgery, inflammation, systemic infection, artificial surface, interventional cardiology, immobility, drugs, pregnancy and fetal loss, and retinopathy, kidney And diabetic complications including neuropathy) can be treated. The compounds can also be used in drug screening assays and other diagnostic and research methods.

III. Pharmaceutical Compositions, Formulations, and Exemplary Routes of Administration and Dosing Considerations Exemplary embodiments of various contemplated agents and pharmaceutical compositions are set forth below.

A. Drug Preparation The compounds of the present invention are useful in the preparation of a medicament for treating various conditions associated with abnormal regulation of cell death, abnormal cell proliferation and hyperproliferation.

  In addition, the compounds are also useful for preparing a medicament for treating other diseases in which the effectiveness of the compound is known or predicted. Such diseases include, but are not limited to, nerve (eg, epilepsy) or neuromuscular diseases. Methods and techniques for preparing pharmaceutical agents of the compounds of the present invention are well known in the art. Exemplary formulations and delivery routes are described below.

  One skilled in the art will recognize that any one or more of the compounds described herein, including many specific embodiments, are prepared by applying standard pharmaceutical manufacturing procedures. . Such agents can be delivered to a subject by using delivery methods well known in the pharmaceutical art.

B. Exemplary Pharmaceutical Compositions and Formulations In some embodiments of the invention, the composition is administered alone, while in some other embodiments, the composition is preferably as defined above. At least one active ingredient / agent with a solid support, with one or more pharmaceutically acceptable carriers, and optionally with another therapeutic agent (e.g., US Provisional Application No. 60 / 607,599 and No. 60 / 641,040, and U.S. Patent Application Nos. 11 / 176,719, 11 / 110,228, 10 / 935,333, 10 / 886,450, 10 / 795,535, 10 / 634,114 10 / 427,211; 10 / 427,212; 10 / 217,878; 09 / 767,283; 09 / 700,101; and related applications; each of which is a whole Presently incorporated herein by reference as a). Each carrier should be “acceptable” in the sense of being compatible with the other ingredients of the formulation and not injurious to the subject.

  Contemplated formulations include oral, rectal, intranasal, topical (including transdermal, buccal and sublingual), vaginal, parenteral (including subcutaneous, intramuscular, intravenous and intradermal) and pulmonary administration. Suitable ones are listed. In some embodiments, the formulation is conveniently present in unit dosage form and is prepared by any method known in the pharmaceutical arts. Such methods include the step of bringing into association the active ingredient with the carrier which constitutes one or more accessory ingredients. In general, the formulations are prepared by uniformly and intimately bringing into association (eg, mixing) the active ingredient with liquid carriers or finely divided solid carriers or both, and then, if necessary, shaping the product.

  Formulations of the present invention suitable for oral administration may be provided as discrete units such as capsules, cachets or tablets, each preferably with a predetermined amount of the active ingredient as a powder or granules; As an aqueous or non-aqueous solution or suspension; or as an oil-in-water liquid emulsion or a water-in-oil liquid emulsion. In another aspect, the active ingredient is provided as a bolus, electuary or paste, and the like.

  In some embodiments, tablets are made by compressing or molding individual agents, including at least one active ingredient and optionally one or more adjuvants / carriers. In some embodiments, compressed tablets can be placed in a suitable machine with freely flowing active ingredients such as powders or granules, optionally binders (e.g., povidone, gelatin, hydroxypropylmethylcellulose), lubricants, inert Prepared by compression with diluents, preservatives, disintegrants (eg sodium starch glycolate, crosslinked povidone, crosslinked sodium carboxymethylcellulose) surface active or dispersing agents. Molded tablets are made by molding in a suitable machine a mixture of the powdered compound (eg, the active ingredient) moistened with an inert liquid diluent. Tablets may optionally be coated or scored, and sustained release of the active ingredient therein, for example, using hydroxypropyl methylcellulose in various proportions to provide the desired release profile Alternatively, it may be formulated to provide controlled release. The tablet may optionally be provided with an enteric coating that is released in the intestinal portion rather than the stomach.

  Formulations suitable for topical administration in the mouth include lozenges containing active ingredients in flavor bases, usually sucrose and acacia or gum tragacanth; active ingredients in inert bases such as gelatin and glycerin, or sucrose and acacia Troches containing; and mouthwashes containing the active ingredients in a suitable liquid carrier.

  A pharmaceutical composition for topical administration according to the invention is optionally formulated as an ointment, cream, suspension, lotion, powder, solution, paste, gel, spray, aerosol or oil. In another aspect, the topical formulation comprises a dressing, such as a patch or bandage or bandage, impregnated with the active ingredient and optionally one or more excipients or diluents. In some embodiments, the topical formulation comprises a compound that enhances absorption or penetration of the active agent through the skin or other affected area. Examples of such skin penetration enhancers include dimethyl sulfoxide (DMSO) and related analogs.

  If desired, the cream-based aqueous phase can be, for example, at least about 30% w / w polyhydric alcohol, i.e., an alcohol having two or more hydroxyl groups, such as propylene glycol, butane-1,3. -Including diol, mannitol, sorbitol, glycerol and propylene glycol and mixtures thereof.

  In some embodiments, the oil phase emulsions of the present invention are composed of known ingredients in a known manner. This phase typically includes only one emulsifier (otherwise known as an emulsifying agent), and in some embodiments, this phase further comprises at least one emulsifier and either fat or oil or both fat and oil. It is also desirable to include a mixture thereof.

  Preferably, a hydrophilic emulsifier is included together with a lipophilic emulsifier that acts as a stabilizer. In some embodiments, it is preferred to include both oil and fat. At the same time, the emulsifier forms a so-called emulsified wax with or without a stabilizer, and the wax together with the oil and / or fat forms a so-called emulsified ointment base that forms the oil dispersed phase of the cream formulation.

  Emulsifiers and emulsion stabilizers suitable for use in the formulations of the present invention include Tween 60, Span 80, cetostearyl alcohol, myristyl alcohol, glyceryl monostearate and sodium lauryl sulfate.

  Since the solubility of the active compound / drug in most oils that may be used in pharmaceutical emulsion formulations is very low, the selection of a suitable oil or fat for the formulation is the desired property (e.g. cosmetic properties) Based on the achievement of. Thus, the cream should preferably be a non-greasy, non-staining aqueous product with a suitable consistency to prevent leakage from tubes or other containers. Linear or branched, mono- or dibasic alkyl esters such as di-isoadipate, isocetyl stearate, propylene glycol diester of coconut fatty acid, isopropyl myristate, decyl oleate, isopropyl palmitate, butyl stearate, 2 A blend of branched chain esters known as -ethylhexyl palmitate or Crodamol CAP may be used, the last three being the preferred esters. These may be used alone or in combination depending on the properties required. High melting point lipids such as white soft paraffin and / or liquid paraffin or other mineral oils can also be used.

  Formulations suitable for topical administration to the eye include eye drops wherein the active ingredient is dissolved or suspended in a suitable carrier, especially an aqueous solvent for the drug.

  Formulations for rectal administration may be presented as a suppository with a suitable base comprising, for example, cocoa butter or a salicylate.

  Formulations suitable for vaginal administration may be provided as pessaries, creams, gels, pastes, foams or spray formulations containing, in addition to the drug, carriers known to be suitable in the art.

  Formulations suitable for intranasal administration wherein the carrier is a solid include coarse powders with a particle size ranging from about 20 to about 500 μm, which are in the manner in which sniffing is ingested, ie, nasally. Administered by rapid inhalation (eg, forced) through the nostrils from a closely held powder container. Other suitable formulations in which the carrier for administration is a liquid include, but are not limited to, nasal sprays, nasal sprays, nebulizer aerosols, including aqueous or oily solutions of the drug.

  Formulations suitable for parenteral administration include aqueous and non-aqueous isotonic sterile injection solutions that may contain antioxidants, buffers, bacteriostats and solutes that render the formulation isotonic with the blood of the intended recipient; and Suspending and thickening agents and aqueous and non-aqueous sterile suspensions that may include liposomes or other particulate systems designed to target the compound to blood components or one or more organs It is done. In some embodiments, the formulation is provided / formulated in single or multiple dose sealed containers, such as ampoules and vials, and a sterile liquid carrier, such as water, is added just prior to use for injection. It may be stored in a freeze-dried (freeze-dried) state that only requires. Extemporaneous injection solutions and suspensions may be prepared from sterile powders, granules and tablets of the kind previously described.

  Preferred single dose formulations are those containing a daily dose or unit, a daily subdose, or an appropriate portion thereof, as cited herein above.

  In addition to the ingredients specifically mentioned above, the formulations of the present invention may include other conventional agents in the art related to the type of formulation in question, such as those suitable for oral administration. It should be understood that other agents such as sweetening, thickening and flavoring agents may be included. It is also contemplated to combine the agents, compositions and methods of the present invention with other suitable compositions and therapies. Still other formulations are optional, such as food additives (suitable sweeteners, flavoring agents, coloring agents, etc.), phytonutrients (e.g. flaxseed oil), minerals (e.g. Ca, Fe, K, etc.), vitamins, And other acceptable compositions (eg, bound linoleic acid), bulking agents, stabilizers, and the like.

C. Exemplary Routes of Administration and Considerations Various delivery systems are known and can be used to administer the therapeutic agents of the invention (eg, the exemplary compounds described above), eg, liposomes Encapsulation, microparticles, microcapsules, receptor-mediated endocytosis, and the like. Delivery methods include, but are not limited to, intraarterial, intramuscular, intravenous, intranasal, and oral routes. In a specific embodiment, it is desirable to administer the pharmaceutical composition of the invention locally to the area in need of treatment; this may be achieved, for example, by local infusion during surgery, injection, or catheter Good, but not limited to these.

  The identified agent can be administered to a subject or individual suspected of, or at risk of developing, the target cell's pathological growth and associated condition. When the agent is administered to a subject such as a mouse, rat or human patient, the agent can be added to a pharmaceutically acceptable carrier and administered systemically or locally to the subject. To identify patients that can be effectively treated, a tissue sample is removed from the patient and the cells are assayed for sensitivity to the drug.

The therapeutic amount is determined by experience and will vary with the condition being treated, the subject being treated and the effectiveness and toxicity of the drug. When delivered to an animal, the method is useful to further confirm the effectiveness of the drug. An example of an animal model is MLR / MpJ-lpr / lpr (“MLR-lpr”) (available from Jackson Laboratories, Bal Harbor, Maine). MLR-lpr mice develop systemic autoimmune disease. Also, as defined herein, other animals may be induced by inducing tumor growth, for example, by inoculating nude mice subcutaneously with about 10 ⁵ to about 10 ⁹ hyperproliferative cancers and target cells. A model can be developed. When a tumor is established, the compounds described herein are administered, for example, by subcutaneous injection around the tumor. Tumor measurements to determine tumor size reduction are made in two dimensions with calipers twice a week. Other animal models may also be used if appropriate. Such animal models for the diseases and conditions are well known in the art.

  In some embodiments, in vivo administration is performed in a single dose, continuously or intermittently throughout treatment. Methods of determining the most effective means and dose of administration are well known to those of skill in the art and will vary with the composition used for treatment, the purpose of the treatment, the target cell being treated, and the subject being treated. . Single or multiple doses are performed at a dosage level and pattern selected by the treating physician.

  Suitable dosage formulations and methods of administering the drug are readily determined by those skilled in the art. Preferably, the compound is administered at about 0.01 mg / kg to about 200 mg / kg, more preferably from about 0.1 mg / kg to about 100 mg / kg, more preferably from about 0.5 mg / kg to about 50 mg / kg. When a compound described herein is administered with another agent (eg, as a sensitizer), the effective amount may be less than when the agent is used alone.

  The pharmaceutical composition can be administered orally, intranasally, parenterally or by inhalation therapy and may be in tablet, lozenge, granule, capsule, pill, ampoule, suppository or aerosol form. They may also be in the form of suspensions, solutions and emulsions of the active ingredient in aqueous or non-aqueous diluents, syrups, granules or powders. In addition to the agents of the present invention, the pharmaceutical compositions can also include other pharmaceutically active compounds or compounds of the present invention.

  More specifically, the agents of the present invention are also referred to herein as active ingredients and for treatment include oral, rectal, intranasal, topical (transdermal, aerosol, buccal and sublingual) Administered by any suitable route including, but not limited to, intravaginal, parenteral (including but not limited to subcutaneous, intramuscular, intravenous and intradermal) and lung. Also good. It will also be appreciated that the preferred route will vary with the condition and age of the recipient and the disease being treated.

  Ideally, the drug should be administered so that a peak concentration of the active compound is achieved at the disease site. This may be accomplished, for example, by intravenous injection of the drug optionally in saline, or administered orally, for example, as a tablet, capsule or syrup containing the active ingredient.

  Desirable blood levels of the drug may be maintained by continuous infusion to provide a therapeutic amount of the active ingredient within the diseased tissue. Effective combinations are intended to provide therapeutic combinations that require a total dose of each component antiviral agent that is lower than that required when individual therapeutic compounds or drugs are used alone. Reduces side effects.

D. Exemplary Co-Administration Routes and Dosing Considerations The present invention also includes methods that involve co-administration of a compound described herein with one or more other active agents. Indeed, another aspect of the present invention is to provide methods for enhancing prior art therapies and pharmaceutical compositions by co-administering a compound of the present invention. In a co-administration procedure, the agents may be administered simultaneously or sequentially. In one embodiment, the compound described herein is administered before the other active agent. The formulation and mode of administration may be any of those described above. In addition, two or more co-administered chemical agents, biological agents, or radiation may each be administered using different modes or different formulations.

  The agent or agents that are co-administered depend on the type of condition being treated. For example, if the condition being treated is cancer, the additional agent can be a chemotherapeutic agent or radiation. If the condition to be treated is an autoimmune disease, the additional agent can be an immunosuppressant or anti-inflammatory agent. If the condition being treated is chronic inflammation, the additional agent can be an anti-inflammatory agent. Additional drugs that are co-administered, such as anti-cancer drugs, immunosuppressive drugs, anti-inflammatory drugs, can be any drug known in the art, including but not limited to those currently in clinical use. . The determination of the appropriate type and dose of radiation therapy is also within the skill of the art or can be determined fairly easily.

  Treatment of various conditions associated with abnormal apoptosis is generally limited by two main factors: (1) development of drug resistance and (2) toxicity of known therapeutic agents. In some cancers, for example, resistance to chemicals and radiation therapy has been shown to be associated with the prevention of apoptosis. Some therapeutic agents have adverse side effects including non-specific lymphocyte toxicity, kidney and bone marrow toxicity.

  The method described herein addresses both these issues. Drug resistance, which increases the dose required to achieve a therapeutic effect, is overcome by co-administering the compounds described herein with known agents. The compounds described herein sensitize target cells to known agents (and vice versa), thus reducing the amount of these agents required to achieve a therapeutic effect.

  The claimed compound sensitization function also addresses problems associated with the toxic effects of known therapies. When known drugs are toxic, in all cases it is desirable to limit the dose administered, especially when the required dose increases due to drug resistance. When the claimed compounds are co-administered with known drugs, they reduce the required dose and thus reduce adverse effects. Furthermore, since the claimed compounds are themselves effective at large doses and are non-toxic, proportionally co-administration of more of these compounds than known toxic therapeutic agents minimizes toxic effects. The desired effect is achieved while suppressing.

IV. Drug Screening In some embodiments of the present invention, the compounds of the present invention, and other potentially useful compounds, are screened for the ability to induce apoptosis.

V. Therapeutic uses In a particularly preferred embodiment, the composition of the present invention comprises a number of conditions (e.g. diseases characterized by abnormal regulation of necrosis and / or apoptotic processes in cells or tissues, abnormal cell proliferation and / or hyperproliferation). Is intended to provide a therapeutic benefit to a patient suffering from any one or more of the diseases characterized by.

  All publications and patents mentioned in the above specification are herein incorporated by reference. Although the invention has been described in connection with specific preferred embodiments, it should be understood that the invention as claimed should not be unduly limited to such specific embodiments. Indeed, various modifications of the described modes for carrying out the invention which are obvious to those skilled in related fields are intended to be within the scope of the following claims.

Claims (12)

A composition comprising the following formula comprising both R and S enantiomers and a racemic mixture:

Wherein A ---- B is selected from the group consisting of N—CH ₂ and C═N;
R1 is hydrogen; halogen; OH; a chemical moiety containing an aryl subgroup; a chemical moiety containing a substituted aryl subgroup; a chemical moiety containing a cycloaliphatic subgroup; a chemical moiety containing a substituted cycloaliphatic subgroup; A chemical moiety containing a group; a chemical moiety containing a substituted heterocyclic subgroup; a chemical moiety containing at least one ester subgroup; a chemical moiety containing at least one ether subgroup; a linear or branched, saturated or unsaturated, substituted Or an aliphatic chain having at least two unsubstituted carbons; a chemical moiety comprising sulfur; a chemical moiety comprising nitrogen; and a chemical moiety wherein R comprises an aryl subgroup; Chemical moieties containing aryl subgroups; chemical moieties containing cycloaliphatic subgroups; chemical moieties containing substituted cycloaliphatic subgroups; chemical moieties containing heterocyclic subgroups; including substituted heterocyclic subgroups A chemical moiety; a linear or branched, saturated or unsaturated, substituted or unsubstituted aliphatic chain having at least two carbons; a chemical moiety comprising at least one ester subgroup; a chemical moiety comprising at least one ether subgroup Selected from the group consisting of chemical moieties containing sulfur; chemical moieties containing nitrogen;
Where R2 is a cyclic group larger than benzene, where greater than benzene includes any chemical group containing 7 or more non-hydrogen atoms;
Wherein R3 is a R3 cyclic structure attached at the 6,7, 7,8 or 8,9 carbon position of the benzodiazepine ring; hydrogen; halogen; OH; chemical moiety containing at least one ester subgroup; at least one ether From a straight chain or branched, saturated or unsaturated, substituted or unsubstituted aliphatic chain having at least two carbons; a sulfur containing chemical moiety; a nitrogen containing chemical moiety; and —OR— Wherein R is a chemical moiety comprising an aryl subgroup; a chemical moiety comprising a substituted aryl subgroup; a chemical moiety comprising a cycloaliphatic subgroup; a chemical moiety comprising a substituted cycloaliphatic subgroup; A chemical moiety containing a group; a chemical moiety containing a substituted heterocyclic subgroup; an aliphatic chain having at least two carbons, linear or branched, saturated or unsaturated, substituted or unsubstituted; containing at least one ester subgroup Selected from the group consisting of: a chemical moiety comprising at least one ether subgroup; a chemical moiety comprising sulfur; a chemical moiety comprising nitrogen;
Wherein at least one of R1 and R3 is a chemical moiety involved in hydrogen bonding;
Wherein R4 is a chemical moiety that causes the benzodiazepine to lack an asymmetric center; and where R5 is a linker group and is present or absent. 2. The composition of claim 1, wherein R1 is selected from the group consisting of:

Wherein, R1 'is halogen, alkyl, substituted alkyl, aryl, substituted aryl; amino; carbonyl; sulfone; sulfonamide; ethers; OH; chemical moieties which contain a nitrogen; chemical moiety containing a sulfur CH _3; straight or An aliphatic chain having at least one carbon that is branched, saturated or unsaturated; an aliphatic chain having at least two carbons that is linear or branched, saturated or unsaturated, and having at least one hydroxy subgroup; An aliphatic chain having at least two carbons that are chain or branched, saturated or unsaturated, and having at least one thiol subgroup; having at least two carbons that are linear or branched, saturated or unsaturated, and having an aldehyde sub Aliphatic chain ending in a group; straight chain or branched, saturated or unsaturated, aliphatic chain having at least one ketone subgroup; straight chain Or an aliphatic chain having at least two branched, saturated or unsaturated carbons and ending with a carboxylic acid subgroup; having at least two carbons that are straight or branched, saturated or unsaturated, and at least one An aliphatic chain having an amide subgroup; an aliphatic chain having at least two carbons that are linear or branched, saturated or unsaturated, and having at least one acyl group; at least one of linear or branched, saturated or unsaturated An aliphatic chain having two carbons and having at least one nitrogen-containing moiety; an aliphatic chain having at least two carbons that are linear or branched, saturated or unsaturated, and having at least one amine subgroup; An aliphatic chain having at least two carbons that are chain or branched, saturated or unsaturated, and having at least one ether subgroup; at least two carbons that are linear or branched, saturated or unsaturated An aliphatic chain having at least one halogen subgroup; a group consisting of an aliphatic chain having at least two carbons, straight or branched, saturated or unsaturated, and having at least one nitronium subgroup More selected. R2 is naphthalene; phenol; 1-naphthalenol; 2-naphthalenol;

2. The composition of claim 1, selected from the group consisting of quinoline, and all aromatic positional isomers. R4 is hydrogen,

2. The composition of claim 1, wherein R4 'is a linear chain or branched, saturated or unsaturated, substituted or unsubstituted aliphatic chain having at least two carbons. 5. The composition of claim 4, wherein the composition is described by the formula:

. 2. The composition of claim 1, wherein the formula is selected from the group consisting of:

. The composition of claim 1, described by the formula:

Wherein R 4 ″ is 1-6 carbons, any one of which is substituted or unsubstituted.   A chemical moiety wherein the R3 cyclic structure comprises an aryl subgroup; a chemical moiety comprising a substituted aryl subgroup; a chemical moiety comprising a cycloaliphatic subgroup; a chemical moiety comprising a substituted cycloaliphatic subgroup; a chemical moiety comprising a heterocyclic subgroup; 2. The composition of claim 1, selected from the group consisting of a chemical moiety comprising a substituted heterocyclic subgroup; a chemical moiety comprising sulfur; a chemical moiety comprising oxygen; and a chemical moiety comprising nitrogen. 9. The composition of claim 8, wherein the compound is selected from the group consisting of:

. 2. The composition of claim 1, wherein the compound is selected from the group consisting of:

.   2. The composition of claim 1, wherein the distance between the chemical moiety involved in hydrogen bonding and the R2 group in three-dimensional space is less than 12 angstroms. ai) the target cell; and
ii) providing a composition according to claim 1; and
b) a method of treating a cell comprising exposing the target cell to a composition.