HK1260272A1 - Aminobenzimidazole derivatives - Google Patents

Description

Aminobenzimidazole derivatives

Cross Reference to Related Applications

This application claims priority to U.S. provisional patent application No. 62/302,781, filed 2016, 3, 2, the contents of which are incorporated herein by reference in their entirety.

Background

Cancer is the second leading cause of death in the united states, and many cancers remain difficult to treat despite new breakthroughs resulting in reduced mortality. In addition, many cancers often develop resistance to current chemotherapy over time. Typical treatments such as chemotherapy, radiation therapy and surgery also cause a wide range of undesirable side effects. Clearly, there is a great need in the art for new compounds and methods that slow the expansion of cancer cells and that can be used to treat cancer.

Disclosure of Invention

The present invention provides a compound which is effective in treating cancer.

It is an object of the present invention to provide a pharmaceutical composition that slows the expansion of cancer cells.

It is an object of the present invention to treat various types of cancer.

The above and other objects can be achieved using a device involving a compound of formula (I):

wherein:

the group represented by X is selected from: H. halogen, -C ₁ -C ₆ Alkyl, aryl, -C ₃ -C ₇ Cycloalkyl, and 3-to 10-membered heterocycle, any of which may be unsubstituted or substituted with one or more of the following: -halogen, -C ₁ -C ₆ Alkyl, -O- (C) ₁ -C ₆ Alkyl), -OH, -CN, -COOR ', -OC (O) R', NHR ', N (R') ₂ A group of, -NHC (O) R ' or-C (O) NHR ', where R ' may be-H or-C ₁ -C ₆ An alkyl group;

the group shown by A is selected from: a bond, -C ₁ -C ₆ Alkyl, or-C ₃ -C ₇ Cycloalkyl groups, any of which may be unsubstituted or substituted with one or more of the following groups: -halogen, -C ₁ -C ₆ Alkyl, -O- (C) ₁ -C ₆ Alkyl), -OH, -CN, -COOR ', -OC (O) R', NHR ', N (R') ₂ A group of, -NHC (O) R ' or-C (O) NHR ', where R ' may be-H or-C ₁ -C ₆ An alkyl group;

the group represented by Y is selected from: H. -C ₁ -C ₆ Alkyl, -C ₃ -C ₇ Cycloalkyl, aryl or a 3 to 10-membered heterocyclic ring, any of which may be unsubstituted or substituted with one or more of the following groups: -halogen, -C ₁ -C ₆ Alkyl, -O- (C) ₁ -C ₆ Alkyl), -OH, -CN, -COOR ', -OC (O) R', NHR ', N (R') ₂ A group of, -NHC (O) R ' or-C (O) NHR ', where R ' may be-H or-C ₁ -C ₆ An alkyl group; and

the group represented by Q is selected from: -halogen, -C ₁ -C ₆ Alkyl, -O- (C) ₁ -C ₆ Alkyl), -OH, -CN, -COOR ', -OC (O) R', NHR ', N (R') ₂ a-NHC (O) R ' or-C (O) NHR ' group, where R ' may be-H or-C ₁ -C ₆ An alkyl group.

In another embodiment, the invention relates to a compound of formula (II):

wherein:

the group represented by X is selected from: H. halogen, -C ₁ -C ₆ Alkyl, aryl, -C ₃ -C ₇ Cycloalkyl, and 3 to 10-membered heterocycle, any of which may be unsubstituted or substituted with one or more of the following: -halogen, -C ₁ -C ₆ Alkyl, -O- (C) ₁ -C ₆ Alkyl), -OH, -CN, -COOR ', -OC (O) R', NHR ', N (R') ₂ A group of, -NHC (O) R ' or-C (O) NHR ', where R ' may be-H or-C ₁ -C ₆ An alkyl group;

a is selected from: -C ₁ -C ₆ Alkyl, or-C ₃ -C ₇ Cycloalkyl groups, any of which may be unsubstituted or substituted with one or more of the following groups: -halogen, -C ₁ -C ₆ Alkyl, -O- (C) ₁ -C ₆ Alkyl), -C ₃ -C ₁₂ Cycloalkyl, 3-to 10-membered heterocycle, aryl, -OH, -CN, -COOR ', -OC (O) R', NHR ', N (R') ₂ A group of, -NHC (O) R ' or-C (O) NHR ', where R ' may be-H or-C ₁ -C ₆ An alkyl group; and

the group represented by Q is selected from: -halogen, -C ₁ -C ₆ Alkyl, -O- (C) ₁ -C ₆ Alkyl), -OH, -CN, -COOR ', -OC (O) R', NHR ', N (R') ₂ A group of, -NHC (O) R ' or-C (O) NHR ', where R ' may be-H or-C ₁ -C ₆ An alkyl group.

The above and other objects can be accomplished using methods that involve the use of an effective amount of a pharmaceutical composition comprising a disclosed compound and (in some aspects of the invention) one or more pharmaceutically acceptable carriers for the reduction of tumor growth.

The above and other objects can be accomplished by a method which involves treating a mammal with an effective amount of a pharmaceutical composition comprising a disclosed compound and (in some aspects of the invention) one or more pharmaceutically acceptable carriers.

The aspects and applications of the inventions presented herein are described in the drawings and detailed description of the inventions. Unless otherwise indicated, the words and phrases in the specification and claims are intended to provide those of ordinary skill in the applicable arts with a plain, ordinary, and accustomed meaning to those words and phrases. The present inventors are fully aware that they can become their own lexicographers if desired

The present inventors have also recognized the normal rules of english grammar. Thus, if a noun, term, or phrase is to be further characterized, specified, or narrowed in some way, such noun, term, or phrase will expressly include additional adjectives, descriptive terms, or other modifiers consistent with the normal rules of english grammar. To the extent that such adjectives, descriptive terms, or modifiers are not used, they are intended to be given the broadest possible meaning.

Furthermore, the inventors are fully aware of 35U.S.C. § 112,to the particular specified standard and application. Thus, use of the words "function," "means" or "step" in the detailed description, drawings, or claims is not intended to indicate that 35u.s.c. § 112 is intended to be referred to in some way,the invention is defined by the specific provisions of. Conversely, if reference is sought 35u.s.c. § 112,the claims will specifically and explicitly state the exact phrase "means for.. or" step for.. and will also recite the word "function" (i.e., will state "to perform an [ insert function ]")]The means for a function of) without reciting in such phrases any structure, material, or act for which such function is supported. Thus, even when the claims recite "a means for performing a function," use "or" use "When "performing a step of a function of.," if the claims also recite any structure, material, or act to support the means or step, or to perform the function, it is expressly intended by the inventors that no reference to 35u.s.c. § 112 is intended by the inventors,the specification of (1). Furthermore, even if reference is made to 35u.s.c. § 112,the definitions are to define the claimed invention, which is intended to be not limited to the specific structures, materials, or acts described in the preferred embodiments, but include any and all structures, materials, or acts for performing the claimed functions, as described in alternate embodiments or forms of the invention, or well-known currently or later-developed equivalent structures, materials, or acts for performing the claimed functions.

Drawings

Figure 1 depicts the effect of treatment with compound ID #1, compound ID #2, or compound ID #3 on tumor volume in a human myeloma xenograft model.

Figure 2 depicts the effect of treatment with compound ID #1, compound ID #2, or compound ID #3 in combination with dexamethasone and pomalidomide (pomalidomide) on tumor volume in a human myeloma xenograft model.

Detailed Description

In the following description, and for purposes of explanation, numerous specific details are set forth in order to provide a thorough understanding of the various aspects of the invention. However, it will be understood by those skilled in the relevant art that the present invention may be practiced without these specific details. In other instances, well-known structures and devices are shown or discussed more generally in order to avoid obscuring the invention. In many cases, the description of the operation is sufficient to enable one to implement various forms of the invention. It should be noted that there are many different and alternative configurations, devices, compositions, and techniques to which the disclosed invention may be applied. The full scope of the invention is not limited to the examples described below.

Herein, the inventors disclose a compound of formula (I):

and a compound of formula (II):

the group represented by X may be H, halogen, -C ₁ -C ₆ Alkyl, aryl, -C ₃ -C ₇ Any of cycloalkyl, or 3-to 10-membered heterocycle, any of which may be unsubstituted or substituted with one or more of the following groups: -halogen, -C ₁ -C ₆ Alkyl, -O- (C) ₁ -C ₆ Alkyl), -OH, -CN, -COOR ', -OC (O) R', NHR ', N (R') ₂ A group of, -NHC (O) R ' or-C (O) NHR ', where R ' may be-H or-C ₁ -C ₆ An alkyl group.

The group shown by A may be a bond, -C ₁ -C ₆ Alkyl, or-C ₃ -C ₇ Any of the cycloalkyl groups, any of which may be unsubstituted or substituted with one or more of the following groups: -halogen, -C ₁ -C ₆ Alkyl, -O- (C) ₁ -C ₆ Alkyl), -OH, -CN, -COOR ', -OC (O) R', NHR ', N (R') ₂ A group of, -NHC (O) R ' or-C (O) NHR ', where R ' may be-H or-C ₁ -C ₆ An alkyl group.

The group represented by Y may be H, -C ₁ -C ₆ Alkyl, -C ₃ -C ₇ Any of cycloalkyl, aryl, or 3 to 10-membered heterocyclic ring, any of which may be unsubstituted or substituted with one or more of the following groups: -halogen, -C ₁ -C ₆ Alkyl, -O- (C) ₁ -C ₆ Alkyl), -OH, -CN, -COOR ', -OC (O) R', NHR ', N (R') ₂ a-NHC (O) R 'or-C (O) NHR' group,wherein R' may be-H or-C ₁ -C ₆ An alkyl group.

The group represented by Q may be H, -halogen, -C ₁ -C ₆ Alkyl, -O- (C) ₁ -C ₆ Alkyl), -OH, -CN, -COOR ', -OC (O) R', NHR ', N (R') ₂ a-NHC (O) R ' or-C (O) NHR ' group, where R ' may be-H or-C ₁ -C ₆ An alkyl group.

-C ₁ -C ₆ Alkyl includes any straight or branched chain, saturated or unsaturated, substituted or unsubstituted hydrocarbon containing from 1 to 6 carbon atoms. -C ₁ -C ₆ Examples of alkyl groups include, but are not limited to, methyl, ethyl, propyl, isopropyl, butyl, sec-butyl, tert-butyl, pentyl, isopentyl, neopentyl, hexyl, isohexyl, neohexyl, ethenyl, propenyl, 1-butenyl, 2-butenyl, 1-pentenyl, 2-pentenyl, 1-hexenyl, 2-hexenyl, 3-hexenyl, ethynyl, pentynyl, 1-butynyl, 2-butynyl, 1-pentynyl, 2-pentynyl, 1-hexynyl, 2-hexynyl, and 3-hexynyl. substituted-C ₁ -C ₆ The alkyl group can include any applicable chemical moiety. May be substituted for any of the above-listed-C ₁ -C ₆ Examples of groups on alkyl include, but are not limited to, the following: halogen, -C ₁ -C ₆ Alkyl, -O- (C) ₁ -C ₆ Alkyl), -OH, -CN, -COOR ', -OC (O) R', -NHR ', N (R') ₂ a-NHC (O) R 'or-C (O) NHR' group. The group represented by R' above may be-H or any-C ₁ -C ₆ An alkyl group.

Aryl includes any unsubstituted or substituted phenyl or naphthyl. Examples of groups that may be substituted onto an aryl group include, but are not limited to: halogen, -C ₁ -C ₆ Alkyl, -O- (C) ₁ -C ₆ Alkyl), -OH, -CN, -COOR ', -OC (O) R', NHR ', N (R') 2, -NHC (O), R ', or-C (O) NEtR'. The group represented by R' may be-H or any-C ₁ -C ₆ An alkyl group.

C ₃ -C ₇ Cycloalkyl includes any 3-, 4-, 5-, 6-, or 7-membered substituted or unsubstituted non-aromatic carbocyclic ring.C ₃ -C ₇ Examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentadienyl, cyclohexyl, cyclohexenyl, cycloheptyl (cyclohexenyl), 1, 3-cyclohexadienyl, -1, 4-cyclohexadienyl, -1, 3-cycloheptadienyl, and-1, 3, 5-cycloheptatrienyl. Can be at C ₃ -C ₇ Examples of substituted groups on cycloalkyl include, but are not limited to: -halogen, -C ₁ -C ₆ Alkyl, -O- (C) ₁ -C ₆ Alkyl), -OH, -CN, -COOR ', -OC (O) R', NHR ', N (R') 2, -NHC (O) R 'or-C (O) NHR' groups. The group represented by R' above includes-H or any unsubstituted-C ₁ -C ₆ Alkyl groups, examples of which are listed above.

Halogen groups include any halogen. Examples include, but are not limited to, -F, -Cl, -Br, or-I.

The heterocyclic ring may be any optionally substituted saturated, unsaturated or aromatic cyclic moiety wherein the cyclic moiety is interrupted by at least one heteroatom selected from oxygen (O), sulfur (S) or nitrogen (N). The heterocyclic ring may be monocyclic or polycyclic. For example, suitable substituents include halogen, halo-C ₁ -C ₆ Alkyl, halo-C ₁ -C ₆ Alkoxy, amino, amidino, acylamino, azido, cyano, guanidino, hydroxy, nitro, nitroso, urea, OS (O) ₂ R、OS(O) ₂ OR、S(O) ₂ OR、S(O) _0-2 R, C (O) OR (where R may be H, C) ₁ -C ₆ Alkyl, aryl OR 3-to 10-membered heterocycle), OP (O) OR ₁ OR ₂ 、P(O)OR ₁ OR ₂ 、SO ₂ NR ₁ R ₂ 、NR ₁ SO ₂ R ₂ 、C(R ₁ )NR ₂ 、C(R ₁ )NOR ₂ (R ₁ And R ₂ May independently be H, C ₁ -C ₆ Alkyl, aryl or 3 to 10 membered heterocyclic ring), NR ₁ C(O)R ₂ 、NR ₁ C(O)OR ₂ 、NR ₃ C(O)NR ₂ R ₁ 、C(O)NR ₁ R ₂ 、OC(O)NR ₁ R ₂ . For these groups, R ₁ 、R ₂ And R ₃ Each independently selected from H, C ₁ -C ₆ Alkyl, aryl or a 3-to 10-membered heterocycle, or R ₁ And R ₂ Together with the atoms to which they are attached form a 3-10 membered heterocyclic ring.

Possible substituents of heterocyclic groups include halogen (Br, Cl, I or F), cyano, nitro, oxo, amino, C _1-4 Alkyl (e.g. CH) ₃ 、C ₂ H ₅ Isopropyl), C _1-4 Alkoxy (e.g., OCH) ₃ 、OC ₂ H ₅ ) Halogen substituted C _1-4 Alkyl (e.g. CF) ₃ 、CHF ₂ ) Of halo C _1-4 Alkoxy (e.g., OCF) ₃ 、OC ₂ F ₅ )、COOH、COO-C _1-4 Alkyl, CO-C _1-4 Alkyl radical, C _1-4 alkyl-S- (e.g., CH) ₃ S、C ₂ H ₅ S), halogeno C _1-4 alkyl-S- (e.g. CF) ₃ S、C ₂ F ₅ S), benzyloxy, and pyrazolyl.

Examples of heterocycles include, but are not limited to, azapinyl, aziridinyl, azetidinyl, azetidinylalkyl, diazepinyl, dithiadiazinyl, dioxaazepinyl, dioxolanyl, dithiazolinyl, dioxazinyl, dithiazolinyl, furanyl, isoxazolyl, isothiazolyl, imidazolyl, morpholinyl, morpholino, oxetanyl, oxadiazolyl, oxiranyl, oxazinyl, oxazolyl, piperazinyl, pyrazinyl, pyridazinyl, pyrimidinyl, piperidinyl, piperidonyl, pyridinyl, pyranyl, pyrazolyl, pyrrolyl, pyrrolidinyl, thiatriazolyl, tetrazolyl, triazolyl, thiazolyl, thienyl, tetrazinyl, thiadiazinyl, triazinyl, thiazinyl, thiafuranonyl, thiadiazolyl, thiazoisoxazolyl, furazolylimidazolyl, thiazoimidazolyl, thiadiazolyl, Thienoisothiazolyl (thienoisothiazolyl), thienothiazolyl, imidazopyrazolyl, cyclopentapyrazolyl, pyrrolopyrrolyl, thienothienyl, thiadiazolopyrimidyl, thiazolothiazinyl, thiazolopyrimidyl, oxazolopyrimidyl, oxazolopyridinyl, benzoxazolyl, benzisothiazolyl, benzothiazolyl, imidazopyrazinyl, purinyl, pyrazolopyrimidyl, imidazopyridinyl, benzimidazolyl, indazolyl, benzoxathienyl (benzoxathiyl), benzodioxolanyl (benzodioxolyl), benzodioxolyl (benzodioxolyl), indolizinyl, dihydroindolyl, isoindolyl, furopyrimidinyl, furopyridinyl, benzofuranyl, isobenzofuranyl, thienopyrimidinyl, thienopyridinyl, benzothiophenyl, cyclopentaoxazinyl, cyclopentafuryl, pyrrolofuranyl, pyrrolopyrrolyl, pyrrolopyrrolylpyridinyl, pyrazolopyridyl, thiazolothiazolyl, and the like, Benzoxazinyl, phenylprophiazinyl, quinazolinyl, naphthyridinyl, quinolinyl, isoquinolinyl, benzopyranyl, pyridopyridazinyl, and pyridopyrimidinyl.

The disclosed compounds and intermediates thereof may exist in different tautomeric forms. Tautomers include any structural isomers of different energies with low mutual transition energy barriers. One example is proton tautomers (prototropic tautomers). In this example, interconversion occurs by migration of protons. Examples of proton tautomers include, but are not limited to, keto-enol and imine-enamine isomerizations. In another example illustrated by the following equation, proton transfer between the nitrogen atoms at the 1-and 3-positions of the benzimidazole ring may occur. As a result, formulae Ia and Ib are tautomeric forms of each other:

the invention also includes any other physicochemical or stereochemical form that the disclosed compounds may take. These forms include diastereomers, racemates, separated enantiomers, hydrated forms, solvated forms, or any other known or undisclosed crystalline, polymorphic crystalline, or amorphous form. Amorphous forms lack a distinguishable crystal lattice and therefore an ordered arrangement of structural units. Many pharmaceutical compounds have an amorphous form. Methods for producing such chemical forms are well known to those skilled in the art.

In some aspects of the invention, the disclosed compounds are in the form of pharmaceutically acceptable salts. Pharmaceutically acceptable salts include any salt derived from an organic or inorganic acid. Examples of such salts include, but are not limited to, the following: hydrobromide, hydrochloride, nitrate, phosphate and sulfate. Organic acid addition salts include, for example, acetate, benzenesulfonate, benzoate, camphorsulfonate, citrate, 2- (4-chlorophenoxy) -2-methylpropionate, 1, 2-ethanedisulfonate, ethanesulfonate, ethylenediaminetetraacetic acid (EDTA) salt, fumarate, glucoheptonate, gluconate, glutamate, N-glycolylarsic acid (N-glycolarsanilic acid) salt, 4-hexylisophthalate, hippurate, 2- (4-hydroxybenzoyl) benzoate, 1-hydroxy-2-naphthoate, 3-hydroxy-2-naphthoate, 2-hydroxyethanesulfonate, lactobionate, N-dodecylsulfate, maleate, malate, mandelate, methanesulfonate, methylsulfate, methosulfate, dihydrogenpropionate, dihydrogenphenate, dihydrogensulfate, dihydrogensulfonate, dihydrogensulfate, and a, Mucate, 2-naphthalenesulfonate, pamoic acid (pamoic acid) salt, pantothenate, phosphatidic acid (4-aminophenyl) phosphonate, picrate, salicylate, stearate, succinate, tannate, tartrate, terephthalate, p-toluenesulfonate, 10-undecenate, or any other salt of an acid now known or not disclosed. One skilled in the art will appreciate that such pharmaceutically acceptable salts can be used in the formulation of pharmacological compositions. These salts can be prepared by reacting the disclosed compounds with a suitable acid in a manner known to those skilled in the art.

The invention also includes aspects of adding a protecting group to the compound. One skilled in the art will recognize that during the synthesis of a complex molecule, one group on the disclosed compounds may happen to interfere with the intended reaction of a second group included on the compound. Temporary masking or protection of the first group may facilitate the desired reaction. Protection involves introducing a protecting group into the group to be protected, carrying out the desired reaction, and removing the protecting group. Removal of the protecting group may be referred to as deprotection. Examples of compounds to be protected in some syntheses include hydroxy, amino, carbonyl, carboxy and mercapto.

Many protecting groups and reagents capable of introducing them into the synthetic process have been and continue to be developed. The protecting group may be produced by any chemical synthesis that selectively attaches a group that is resistant to certain reagents to the chemical group to be protected without significantly affecting any other chemical group in the molecule, remains stable throughout the synthesis, and can be removed by conditions that do not adversely react with the protected group and any other chemical group in the molecule. Multiple protecting groups may be added throughout the synthesis, and one skilled in the art will be able to develop strategies for the specific addition and removal of protecting groups to and from the group to be protected.

Protecting groups, reagents for adding such groups, preparation of such reagents, protection and deprotection strategies under a variety of conditions, including complex syntheses with complementary protecting groups, are well known in the art. Non-limiting examples of all of these can be found in Green et al, Protective Groups in Organic Chemistry, second edition, (Wiley 1991) and Harrison et al, Complex of Synthetic Organic Methods, volumes 1-8 (Wiley, 1971-1996), the entire contents of which are incorporated herein by reference.

The racemates, individual enantiomers, or diastereomers of the disclosed compounds can be prepared by any method, now known or not disclosed, by specific synthesis or resolution. For example, the disclosed compounds can be resolved into enantiomers by forming diastereomeric pairs by salt formation using an optically active acid. The enantiomers are crystallized stepwise, yielding the free base. In another example, enantiomers may be separated by chromatography. Such chromatography may be any suitable method now known or as yet undisclosed, suitable for separating enantiomers, such as HPLC on a chiral column.

The invention also includes pharmaceutical compositions comprising one of the disclosed compounds as an ingredient. These pharmaceutical compositions may take any necessary physical form, depending on a number of factors, including the desired method of administration and the physicochemical and stereochemical forms taken by the disclosed compounds or pharmaceutically acceptable salts of the compounds. These physical forms include solids, liquids, gases, sols, gels, aerosols, or any other physical form now known or not yet disclosed. The concept of a pharmaceutical composition comprising the disclosed compound also includes the disclosed compound or a pharmaceutically acceptable salt thereof without any other additives. The physical form of the invention may affect the route of administration, and one skilled in the art will know to select a route of administration that takes into account the physical form of the compound and the condition to be treated. Pharmaceutical compositions comprising one of the disclosed compounds can be prepared using methods well known in the art of pharmacy. A pharmaceutical composition comprising one of the disclosed compounds can comprise a second effective compound that is different in chemical formula from the disclosed compound. The second effective compound may have the same or similar molecular target as the target, or it may act upstream or downstream of the molecular target of the disclosed compound in relation to one or more biochemical pathways.

Pharmaceutical compositions comprising one of the disclosed compounds include materials that can alter the physical form of the dosage unit. In one non-limiting example, the composition includes a material that forms a coating of the constraining compound. Materials that can be used for such coatings include, for example, sugar, shellac, gelatin, or any other inert coating agent.

Pharmaceutical compositions comprising one of the disclosed compounds can be prepared as a gas or aerosol. Aerosols include a variety of systems, including gels and pressurized packs. Delivery of such forms of the compositions may include propelling a pharmaceutical composition containing the disclosed compounds through the use of liquefied or other compressed gases or through a suitable pump system. The aerosol may be delivered in a single phase, two phase or three phase system.

In some aspects of the invention, a pharmaceutical composition of one of the disclosed compounds is in the form of a solvate. Such solvates are prepared by dissolving the disclosed compounds in a pharmaceutically acceptable solvent. A pharmaceutically acceptable solvent includes any mixture of more than one solvent. These solvents can include pyridine, chloroform, propan-1-ol, ethyl oleate, ethyl lactate, ethylene oxide, water, ethanol, and any other solvent that delivers sufficient amounts of the disclosed compounds to treat pain without the serious complications that arise from the use of solvents in most patients.

Pharmaceutical compositions comprising one of the disclosed compounds may also comprise a pharmaceutically acceptable carrier. Carriers include any substance that can be administered with the disclosed compounds for the purpose of facilitating, assisting or aiding in the administration or other delivery of the compounds. Carriers include any liquid, solid, semi-solid, gel, aerosol, or any other substance that can be combined with the disclosed compounds to aid in their administration. Examples include diluents, adjuvants, excipients, water, oils (including petroleum, animal, vegetable or synthetic oils). Such carriers include particles such as tablets or powders, liquids such as oral syrups or injectable liquids, and inhalable aerosols. Other examples include saline, gum arabic, gelatin, starch paste, talc, keratin, colloidal silica, and urea. These carriers may also include binders such as ethyl cellulose, carboxymethyl cellulose, microcrystalline cellulose, or gelatin; excipients such as starch, lactose or dextrin; disintegrating agents such as alginic acid, sodium alginate, Primogel and corn starch; lubricants such as magnesium stearate or Sterotex; glidants such as colloidal silicon dioxide; sweetening agents such as sucrose or saccharin; a flavoring agent such as peppermint, methyl salicylate, or orange flavoring; or a colorant. Other examples of carriers include polyethylene glycol, cyclodextrin, oil, or any other similar liquid carrier that can be formulated into a capsule. Other examples of carriers include sterile diluents such as water for injection, saline solution, physiological saline, ringer's solution, isotonic sodium chloride, fixed oils such as synthetic mono-or diglycerides, polyethylene glycols, glycerol, cyclodextrins, propylene glycol or other solvents; antibacterial agents such as benzyl alcohol or methyl paraben; antioxidants, such as ascorbic acid or sodium bisulfite; chelating agents, such as ethylenediaminetetraacetic acid; buffers such as acetates, citrates or phosphates, and agents for adjusting the osmotic pressure such as sodium chloride or dextrose, thickeners, lubricants and colorants.

Pharmaceutical compositions comprising one of the disclosed compounds may take any of a number of formulations, depending on the physicochemical form and type of administration of the composition. These forms include solutions, suspensions, emulsions, tablets, pills (pils), pellets (pelles), capsules containing liquids, powders, sustained release formulations, targeted release formulations, lyophilizates, suppositories, emulsions, aerosols, sprays, granules, powders, syrups, elixirs, or any other formulation now known or not disclosed. Other examples of suitable Pharmaceutical carriers are described in "Remington's Pharmaceutical Sciences" by e.w. martin, which is incorporated herein by reference in its entirety.

Methods of administration include, but are not limited to, oral administration and parenteral administration. Parenteral administration includes, but is not limited to, intradermal, intramuscular, intraperitoneal, intravenous, subcutaneous, intranasal, epidural, sublingual, intramucosal, intracerebral, intraventricular, intrathecal, intravaginal, transdermal, rectal, by inhalation, or topical administration to the ear, nose, eye, or skin. Other methods of administration include, but are not limited to, infusion techniques, including infusion or bolus injection, absorption through epithelial or mucocutaneous linings (lines), such as the oral mucosa, rectum and intestinal mucosa. Compositions for parenteral administration may be enclosed in ampoules, disposable syringes or multiple dose vials made of glass, plastic or other material.

Administration may be systemic or local. Topical administration is the administration of the disclosed compounds to the area in need of treatment. Examples include local infusion during surgery, external administration (topical applications), local injection, or administration through a catheter, suppository, or implant. Administration can be by direct injection into the site (or pre-site) of the cancer, tumor or precancerous tissue or the central nervous system by any suitable route, including intraventricular and intrathecal injection. Intraventricular injection may be assisted by an intraventricular catheter, for example connected to a reservoir such as an Ommaya reservoir. Pulmonary administration can be accomplished by any of a number of methods known in the art. Examples include the use of an inhaler or nebulizer, aerosol-containing formulations, or by infusion in fluorocarbons or synthetic lung surfactants. The disclosed compounds may be delivered in the context of vesicles (e.g., liposomes or any other natural or synthetic vesicle).

Pharmaceutical compositions formulated for administration by injection can be prepared by dissolving the disclosed compounds with water to form a solution. In addition, surfactants may be added to promote the formation of a homogeneous solution or suspension. Surfactants include any complex capable of non-covalent interaction with the disclosed compounds to facilitate dissolution or uniform suspension of the compounds.

Pharmaceutical compositions comprising one of the disclosed compounds can be prepared in a form that facilitates topical or transdermal administration. These formulations may be in the form of solutions, emulsions, ointments, gel matrices, transdermal patches or iontophoresis devices. Examples of bases for use in such compositions include petrolatum, lanolin, polyethylene glycols, beeswax, mineral oil, diluents such as water and alcohols, and emulsifiers and stabilizers, thickeners, or any other suitable base now known or not disclosed.

Cancer cells include any cell derived from a tumor, neoplasia (neoplasms), cancer, precancerous lesion, cell line, or any other cell source that is ultimately capable of potentially unlimited expansion and growth. Cancer cells may be derived from naturally occurring sources or may be artificially produced. When placed in an animal host, cancer cells can also invade other tissues and metastasize. Cancer cells also include any malignant cells that invade other tissues and/or metastasize. One or more cancer cells in the context of an organism may also be referred to as a cancer, tumor, neoplasia, growth, malignancy, or any other term used in the art to describe a cell of a cancer state.

Expansion of cancer cells includes any process that results in an increase in the number of individual cells derived from the cancer cells. The expansion of cancer cells can be caused by mitosis, proliferation, or any other form of expansion of cancer cells, whether in vitro or in vivo. The expansion of cancer cells further includes invasion and metastasis. Cancer cells may be physically close to cancer cells from the same clone or from a different clone that may or may not be genetically identical thereto. This aggregation may take the form of colonies (colony), tumors or metastases, any of which may occur in vivo or in vitro. Slowing the expansion of cancer cells can be achieved by inhibiting cellular processes that promote expansion or by causing cellular processes that inhibit expansion. Processes that inhibit expansion include processes that slow mitosis and processes that promote cell senescence or cell death. Examples of specific processes that inhibit amplification include caspase-dependent and independent pathways, autophagy, necrosis, apoptosis, and mitochondria-dependent and independent processes, and also include any such processes yet to be disclosed.

The addition of the pharmaceutical composition to the cancer cells includes all activities that effect the pharmaceutical composition on the cancer cells. The type of addition selected will depend on whether the cancer cell is in vivo, ex vivo or in vitro, the physical or chemical properties of the pharmaceutical composition, and the effect of the composition on the cancer cell. Non-limiting examples of additions include adding a solution comprising the pharmaceutical composition to a tissue culture medium in which cancer cells are growing in vitro; the pharmaceutical composition may be administered to the animal by any method, including intravenous, oral, parenteral, or any other method of administration; or activation or inhibition of cells, which in turn are responsible for cancer cells such as immune cells (e.g., macrophages and CD 8) ⁺ T cells) or endothelial cells that can differentiate into vascular structures during angiogenesis or vasculogenesis.

Determining an effective amount of the disclosed compounds is within the ability of those skilled in the art, particularly in light of the detailed disclosure provided herein. The effective amount of a pharmaceutical composition to achieve a particular purpose, as well as its toxicity, excretion, and overall tolerability, can be determined in cell cultures or experimental animals by pharmaceutical and toxicological methods now known to those of skill in the art, or by any similar method not yet disclosed. One example is the in vitro determination of the IC of a pharmaceutical composition in a cell line or target molecule ₅₀ (half maximal inhibitory concentration). Another example is the determination of LD for a pharmaceutical composition in an experimental animal ₅₀ (lethal dose resulting in death of 50% of the test animals). For determining an effective amountThe exact technique will depend on factors such as the type and physical/chemical properties of the pharmaceutical composition, the properties to be tested, and whether the test is to be performed in vitro or in vivo. Determination of an effective amount of a pharmaceutical composition is well known to those skilled in the art and will use data obtained from any of the tests to perform the assay. Determining an effective amount of the disclosed compounds for addition to cancer cells also includes determining a therapeutically effective amount, including a range of effective dosages of the formulation for use in vivo, including in humans.

Treatment is contemplated in biological entities including, but not limited to, mammals (particularly humans) and other mammals of economic or social importance, including mammals in endangered states. Further examples include livestock or other animals, typically raised for human consumption, and domestic companion animals.

Toxicity and therapeutic efficacy of the pharmaceutical compositions can be determined by standard pharmaceutical procedures in cell cultures or animals. Examples include determining the IC of a test compound ₅₀ (half maximal inhibitory concentration) and LD ₅₀ (lethal dose resulting in death of 50% of the test animals). The data obtained from these cell culture assays and animal studies can be used to formulate a range of doses for use in humans. The dosage may vary depending on the dosage form employed and the route of administration utilized.

An effective amount of one of the disclosed compounds that results in slow expansion and contraction of cancer cells preferably results in a concentration at or near the target tissue that is effective to slow cell expansion in tumor cells, but has little effect on non-tumor cells, including non-tumor cells exposed to radiation or recognized chemotherapeutic chemicals. The concentration at which these effects occur can be determined using, for example, apoptosis markers such as the apoptosis index and/or caspase activity in vitro or in vivo.

Treating a condition is the practice of any method, process, or procedure that aims to halt, inhibit, slow, or reverse the progression of a disease, disorder, or condition, to substantially alleviate clinical symptoms of a disease, disorder, or condition, or to substantially prevent the appearance of clinical symptoms of a disease, disorder, or condition, up to and including returning the diseased entity to a state prior to the development of the disease.

Adding a therapeutically effective amount of one of the disclosed compounds includes any method of administering the compound. Administration of the disclosed compounds can include single or multiple administrations of any of a number of pharmaceutical compositions comprising the disclosed compounds as active ingredients. Examples include a single administration of a sustained release composition, a course of treatment involving multiple treatments on a regular or irregular basis, multiple administrations for a period of time until a reduction in the disease state is achieved, a prophylactic treatment applied prior to the onset of symptoms, or any other dosing regimen known to those of skill in the art or yet to be disclosed as a potentially effective regimen. The final dosing regimen, including the regularity of administration and the mode of administration, will depend on any of a number of factors, including but not limited to the subject being treated, the severity of the affliction, the mode of administration, the stage of development of the disease, the presence of one or more other conditions, such as pregnancy, infancy, or the presence of one or more other diseases, or any other factor affecting the choice of mode of administration, the dosage administered, and the time period over which the dosage is administered, now known or not disclosed.

A pharmaceutical composition comprising one of the disclosed compounds can be administered prior to, concurrently with, or subsequent to the administration of a second pharmaceutical composition that may or may not comprise the compound. If the compositions are administered simultaneously, they are administered within one minute of each other. If not administered simultaneously, the second pharmaceutical composition may be administered one or more minutes, hours, days, weeks or months before or after the pharmaceutical composition comprising the compound. Alternatively, combinations of pharmaceutical compositions may be administered cyclically. Cycling therapy involves administering one or more pharmaceutical compositions for a period of time, then administering one or more different pharmaceutical compositions for a period of time and repeating this sequential administration to reduce the development of resistance to the one or more compositions, to avoid or reduce side effects of the one or more compositions, and/or to improve the efficacy of the treatment.

The invention also includes kits that facilitate administration of one of the disclosed compounds to a diseased entity. Examples of such kits include one or more unit doses of the compound. Unit doses are enclosed in preferably sterile containers and contain the disclosed compounds and a pharmaceutically acceptable carrier. In another aspect, the unit dose comprises a lyophilizate of one or more compounds. In this aspect of the invention, the kit may comprise another, preferably sterile, container which encloses a solution capable of dissolving the lyophilizate. However, such a solution need not be included in the kit, and can be obtained separately from the lyophilizate. In another aspect, the kit may include one or more devices for administering a unit dose or pharmaceutical composition used in combination with a compound. Examples of such devices include, but are not limited to, syringes, drip bags, patches, or enemas. In some aspects of the invention, the device comprises a container enclosing the unit dose.

Pharmaceutical compositions comprising one of the disclosed compounds are useful in methods of treating cancer. The methods involve administering to a mammal, preferably a mammal that has been diagnosed with cancer, a therapeutic amount of a pharmaceutical composition comprising a disclosed compound and/or a pharmaceutically acceptable salt thereof.

Therapeutic amounts also include preventing the progression of the cancer to a neoplastic, malignant, or metastatic state. Such prophylactic use is suggested in conditions known or suspected to have previously developed neoplasia (neoplasma) or cancer, particularly where non-neoplastic cell growth consisting of hyperplasia, metaplasia or, particularly, dysplasia has occurred (for a review of such abnormal growth conditions, see robblins and Angell, 1976, Basic Pathology, second edition, WB Saunders co., philidelphia, pp.68-79). Proliferation is a form of controlled cell proliferation that involves an increase in the number of cells in a tissue or organ without significant changes in structure or activity. For example, endometrial hyperplasia usually precedes endometrial cancer, while precancerous colon polyps usually turn into cancerous lesions. Tissue deformation is a form of controlled cell growth in which one adult or fully differentiated cell replaces another type of adult cell. Tissue deformation may occur in epithelial cells or connective tissue cells. Typical metaplasia includes metaplastic epithelial cells that are somewhat disordered. Dysplasia is often a precursor to cancer, primarily found in epithelial cells; it is the most disorderly form of growth in non-tumor cells, involving loss of single cell uniformity and cell structural orientation. Dysplastic cells typically have abnormally large, deeply stained nuclei and exhibit polymorphism. Dysplasia characteristically occurs where there is chronic irritation or inflammation, and is often found in the cervix, respiratory tract, oral cavity, and gallbladder.

As an alternative or in addition to the presence of abnormal cell growth characterized by hyperplasia, metaplasia or dysplasia, the presence of one or more characteristics of a transformed or malignant phenotype, displayed in vivo or displayed in vitro by a patient-derived cell sample, may indicate the need for prophylactic/therapeutic administration of a pharmaceutical composition comprising the compound. Such characteristics of the transformed phenotype include morphological changes, weaker matrix attachment, loss of contact inhibition, loss of anchorage dependence, protease release, increased sugar transport, decreased serum demand, fetal antigen expression, disappearance of 250,000 dalton cell surface proteins, etc. (see above, pages 84-90, characteristics associated with the transformed or malignant phenotype). Further examples include leukoplakia (benign apparent hyperplasia or dysplastic lesions of the epithelium) or Bowen's disease (carcinoma in situ), which is a pre-neoplastic lesion, indicating the desirability of prophylactic intervention. In another example, fibrocystic disease, including cystic hyperplasia, breast dysplasia, adenopathy, or benign epithelial hyperplasia, indicates the desirability of prophylactic intervention.

In some aspects of the invention, the use of the disclosed compounds may be determined by one or more physical factors, such as tumor size and grade or one or more molecular markers and/or expression signatures indicative of prognosis and possible response to treatment with the compounds. For example, determination of Estrogen (ER) and Progesterone (PR) steroid hormone receptor status has become a routine procedure for the evaluation of breast cancer patients. See, e.g., fitzgibbs et al, arch.pathol.lab.med.124: 966-78, 2000. Hormone receptor positive tumors are more likely to respond to hormone therapy and often also do not grow as actively, resulting in ER ⁺ /PR ⁺ The prognosis of patients with tumors is better. In another example, overexpression of human epidermal growth factor receptor 2(HER-2/neu), a transmembrane tyrosine kinase receptor protein, is associated with poor prognosis of breast cancer (see, e.g., Ross et al, The Oncoloist 8: 307-25, 2003), and levels of HER-2 expression in breast tumors are used to predict The anti-HER-2 monoclonal antibody therapeutic trastuzumab (R-2-T-R)Genentech, southern francisco, CA).

In another aspect of the invention, a diseased entity exhibits one or more predisposing factors for malignancy that may be treated by administering a pharmaceutical composition comprising the compound. These predisposing factors include, but are not limited to, chromosomal translocations associated with malignancies, such as Philadelphia chromosome for chronic myelogenous leukemia and t for follicular lymphoma (14; 18); indicating the incidence of polyposis or gardner syndrome of colon cancer; benign monoclonal gammopathy indicative of multiple myeloma; relativity to persons suffering from or currently suffering from cancer or precancerous conditions; exposure to carcinogens; or any other predisposing factor now known or yet to be disclosed that indicates an increased incidence of cancer.

The invention also includes methods of treating cancer comprising combination therapy, wherein the combination therapy comprises administering a pharmaceutical composition comprising one of the disclosed compounds and another modality of treatment. Such treatment modalities include, but are not limited to, radiation therapy, chemotherapy, surgery, immunotherapy, cancer vaccines, radioimmunotherapy, treatment with pharmaceutical compositions other than those comprising the disclosed compounds, or any other method now known or not disclosed that is effective in treating cancer in combination with the disclosed compounds. The combination therapy may act synergistically. That is, the combination of the two therapies is more effective than either therapy administered alone. This leads to a situation where lower doses of both treatment modalities can be effectively used. This in turn reduces toxicity and side effects (if any) associated with either mode of administration without reducing efficacy.

In another aspect of the invention, a pharmaceutical composition comprising one of the disclosed compounds is administered in combination with a therapeutically effective amount of radiation therapy. Radiation therapy can be administered simultaneously, prior to, or after a pharmaceutical composition comprising the compound. Radiation therapy may be additive or synergistic with pharmaceutical compositions comprising the compounds. This particular aspect of the invention is most effective in cancers known to respond to radiation therapy. Cancers known to be responsive to radiation therapy include, but are not limited to, non-hodgkin's lymphoma, hodgkin's disease, ewing's sarcoma, testicular cancer, prostate cancer, ovarian cancer, bladder cancer, laryngeal cancer, cervical cancer, nasopharyngeal cancer, breast cancer, colon cancer, pancreatic cancer, head and neck cancer, esophageal cancer, rectal cancer, small cell lung cancer, non-small cell lung cancer, brain tumors, other CNS tumors, or any other such tumor now known or not disclosed.

Examples of pharmaceutical components that can be used in combination with one of the disclosed compounds include nucleic acid binding components such as cis-diammineplatinum (II) dichloride (cisplatin), doxorubicin, 5-fluorouracil, paclitaxel, and topoisomerase inhibitors such as etoposide, teniposide, irinotecan, and topotecan. Other pharmaceutical ingredients include antiemetic ingredients such as metoclopramide (metoclopramide), domperidone, prochlorperazine, promethazine, chlorpromazine, trimethobenzamide, ondansetron, granisetron, hydroxyzine, acetylleucine monoethanolamine (acetoleucinemonelamine), azimilide, azasetron, benzquinamine, biacetadine (bietanautine), brompride, buclizine, clopride, cyclizine, dimenhydrinate, dichlorphenadol (diphenidol), dolasetron, meclizine, mesalamine, metopimazine, naloxondol, oxypendyl, pipamazine, scopolamine, sulpiride, tetrahydrocannabinol, thiethylperazine, thioproperazine, and tropisetron.

Other examples of pharmaceutical components that can be used in combination with a pharmaceutical composition comprising one of the disclosed compounds are hematopoietic colony stimulating factors. Examples of hematopoietic colony stimulating factors include, but are not limited to, filgrastim, sargrastim, morastim, and alfa-epoetin. Alternatively, a pharmaceutical composition comprising one of the disclosed compounds may be used in combination with an anxiolytic. Examples of anxiolytics include, but are not limited to, buspirone and benzodiazepines such as diazepam, lorazepam, oxazepam, chlordiazepam (chlorazepate), clonazepam, methotrexate (chlordiazepam), and alprazolam.

Pharmaceutical compositions that can be used in combination with a pharmaceutical composition comprising one of the disclosed compounds can include an analgesic. Such agents may be opioid analgesics or non-opioid analgesics. Non-limiting examples of opioid analgesics include morphine, heroin, hydromorphone, hydrocodone, oxymorphone, oxycodone, metodone, apomorphine, normorphine, etorphine, buprenorphine, meperidine, loperamide, anileridine, isoxepinine (ethoheptazine), piminodine (piminidine), betadine, diphenoxylate, fentanyl, sufentanil, alfentanil, remifentanil, levorphanol, dextromethorphan, finazocine, pentazocine, cyclazocine, methadone, isometholone, and propoxyphene. Suitable non-opioid analgesics include, but are not limited to, aspirin, celecoxib, rofecoxib, diclofenac (diclofinac), diflunisal (diflusinal), etodolac (etodolac), fenoprofen, flurbiprofen, ibuprofen, ketoprofen, indomethacin, ketorolac, meclofenoxate, mefenamic acid (mefanamic acid), nabumetone, naproxen, piroxicam, sulindac, or any other analgesic now known or not disclosed.

In other aspects of the invention, a pharmaceutical composition comprising one of the disclosed compounds may be used in combination with a method involving ex vivo treatment of cancer. An example of such a treatment is autologous stem cell transplantation. In this method, autologous hematopoietic stem cells of the diseased entity are harvested and all cancer cells are eliminated. A therapeutic amount of a pharmaceutical composition comprising one of the disclosed compounds can then be administered to a patient prior to restoring solid bone marrow by addition of the patient's own or donor stem cells.

Cancers that can be treated by a pharmaceutical composition (including one of the disclosed compounds) alone or in combination with another treatment modality include solid tumors, such as fibrosarcoma, myxosarcoma, liposarcoma, chondrosarcoma, osteogenic sarcoma, chordoma, angiosarcoma, endotheliosarcoma, lymphangiosarcoma, lymphangioendotheliosarcoma (lymphangioendotheliosarcoma), synovioma, mesothelioma, ewing's tumor, leiomyosarcoma, rhabdomyosarcoma, colon cancer, colorectal cancer, kidney cancer, pancreatic cancer, bone cancer, breast cancer, ovarian cancer, prostate cancer, esophageal cancer, stomach cancer, oral cancer, nasal cancer, pharyngeal cancer, squamous cell cancer, basal cell cancer, adenocarcinoma, sweat gland cancer, papillary adenocarcinoma, cystadenocarcinoma, medullary cancer, bronchial cancer, renal cell cancer, liver cancer, bile duct cancer, choriocarcinoma, seminoma, embryonal cancer, embryonic cancer, choriocarcinoma, and the like cancers, Wilms' tumor, cervical cancer, uterine cancer, testicular cancer, small cell lung cancer, bladder cancer, lung cancer, epithelial cancer, glioma, glioblastoma multiforme (glioblastomas multiforme), astrocytoma, medulloblastoma, craniopharyngioma, ependymoma, pinealoma, hemangioblastoma, acoustic neuroma, oligodendroglioma, meningioma, skin cancer, melanoma, neuroblastoma, and retinoblastoma.

Other cancers that may be treated by pharmaceutical compositions comprising the disclosed compounds include blood-borne cancers such as acute lymphoblastic leukemia ("ALL"), acute lymphoblastic B-cell leukemia, acute lymphoblastic T-cell leukemia, acute myeloblastic leukemia ("AML"), acute promyelocytic leukemia ("APL"), acute monocytic leukemia, acute erythroleukemic leukemia, acute megakaryoblastic leukemia, acute myelomonocytic leukemia, acute non-lymphocytic leukemia, acute undifferentiated leukemia, chronic myelocytic leukemia ("CML"), chronic lymphocytic leukemia ("CLL"), hairy cell leukemia, multiple myeloma, lymphoblastic leukemia, myeloid leukemia, lymphocytic leukemia, granulocytic leukemia, hodgkin's disease, non-hodgkin's lymphoma, Waldenstrom's macroglobulinemia, heavy chain disease, and polycythemia.

The following are examples that represent various aspects of the present invention. These examples should not be construed as limiting the scope of the disclosure. Alternative mechanistic pathways and similar structures within the scope of the invention will be apparent to those skilled in the art.

Examples

The elements and acts in the examples are intended to be illustrative of the invention for simplicity and clarity and have not necessarily been presented in any particular order or implementation. The examples are also intended to establish the ownership of the invention by the inventors.

Example 1 exemplary Compounds of formula (I) or formula (II)

4- ((1- (cyclohexylmethyl) -1H-benzo [ d ] imidazol-2-yl) amino) -N-hydroxybenzamide. ID #1

4- ((1-cyclohexyl-1H-benzo [ d ] imidazol-2-yl) amino) -N-hydroxybenzamide. ID #2

4- ((1-cycloheptyl-1H-benzo [ d ] imidazol-2-yl) amino) -N-hydroxybenzamide. ID #3

4- ((1- ((1-fluorocyclohexyl) methyl) -1H-benzo [ d ] imidazol-2-yl) amino) -N-hydroxybenzamide. ID #4

4- ((1- (cyclopentylmethyl) -1H-benzo [ d ] imidazol-2-yl) amino) -N-hydroxybenzamide. ID #5

4- ((1- (2-cyclopentylethyl) -1H-benzo [ d ] imidazol-2-yl) amino) -N-hydroxybenzamide. ID #6

4- ((1- ((4, 4-Difluorocyclohexyl) methyl) -1H-benzo [ d ] imidazol-2-yl) amino) -N-hydroxybenzamide ID #7

4- ((1- ((4, 4-difluorocyclohexyl) methyl) -5-fluoro-1H-benzo [ d ] imidazol-2-yl) amino) -N-hydroxybenzamide. ID #8

N-hydroxy-4- ((1- ((tetrahydro-2H-pyran-4-yl) methyl) -1H-benzo [ d ] imidazol-2-yl) amino) benzamide. ID #9

4- ((5-fluoro-1- ((tetrahydro-2H-pyran-4-yl) methyl) -1H-benzo [ d ] imidazol-2-yl) amino) -N-hydroxybenzamide. ID #10

Example 2 cell viability assay Using MM1.S cells

Cell viability was used to assess cytotoxicity and the effect of compounds on cell proliferation in the presence of different concentrations of the above compounds at different time points. Chemolysis disclosed in MM1.S cell lineIC of compound ₅₀ The (or percent activity) data are summarized in table 1.

Cell viability assay-by from Promega (Madison, Wis.)Cell viability assay measures cell viability.The luminocyte viability assay is a homogeneous method for determining the number of viable cells in culture based on the quantification of the presence of ATP, which is indicative of the presence of metabolically active cells. After treatment, theAdded to the treatment wells and incubated at 37 ℃. The luminescence values were measured using a Molecular Devices Spectramax microplate reader.

Single reagent study-cells were grown to 70% confluence, trypsinized, counted, and plated in 96-well flat-bottom plates to a final concentration of 2.5 × 10 ³ -5x10 ³ Cells/well (day 0). Cells were incubated in growth medium for 24 hours. Treatment with test or standard reagents was started on day 1 and continued for 72 hours. At the 72 hour time point, the treatment containing the medium was removed. As described above, byCell viability assays to quantify viable cell number. The results of these studies were used to calculate the IC of each compound ₅₀ Values (concentration of drug that inhibited cell growth by 50% of control).

Data collection-data from each experiment was collected for single reagent and combination studies and expressed as% cell growth using the following calculation:

growth of cells% _Testing /f _Carrier )x 100

Wherein f is _Testing Is the fluorescence of the sample to be tested, and f _Carrier Is the fluorescence of the carrier used to dissolve the drug. Using Prism 6 softPart (GraphPad), dose response plot and IC were generated using the following equations ₅₀ The value:

Y＝(Top-bottom)

(1+10 ^{((logIC50-X) -slope)} )

Wherein X is the log of the concentration and Y is the response. Y rises from the bottom to the top in an S-shape.

TABLE 1 IC of example Compounds in MM1.S cells ₅₀

Example 3 cell viability assay Using various cell lines

Compounds ID #1 (compound of formula (I)) and compound ID #3 (compound of formula (II)) were tested for inhibition of cancer cell proliferation. Cell viability was used to assess cytotoxicity and the effect of compounds on cell proliferation in the presence of different concentrations of compound ID #1 and compound ID #3 at different time points. 50% Inhibitory Concentration (IC) of Compound ₅₀ ) The data are summarized in table 2. The data clearly show the surprising and unexpected increased anticancer activity associated with compound ID #1 compared to compound ID # 3.

Cell viability assay-by from Promega (Madison, Wis.)Cell viability assay measures cell viability.The luminocyte viability assay is a homogeneous method for determining the number of viable cells in culture based on the quantification of the presence of ATP, which is indicative of the presence of metabolically active cells. After treatment, theAdded to the treatment wells and incubated at 37 ℃. The luminescence values were measured using a Molecular Devices Spectramax microplate reader.

Single reagent study-cells were grown to 70% confluence, trypsinized, counted, and plated in 96-well flat-bottom plates to a final concentration of 2.5 × 10 ³ -5x10 ³ Cells/well (day 0). Cells were incubated in growth medium for 24 hours. Treatment with test agent was started on day 1 and continued for 72 hours. At the 72 hour time point, the medium containing the treatment was removed. As described above, byCell viability assays to quantify viable cell number. Experiments were performed using triplicate concentrations to determine growth inhibitory activity. The results of these studies were used to calculate the IC of each compound ₅₀ Values (concentration of drug that inhibited cell growth by 50% of control).

Data collection-for single agent and combination studies, data from each experiment was collected and expressed as% cell growth using the following calculation:

growth of cells% _Testing /f _Carrier )x 100

Wherein f is _Testing Is the fluorescence of the sample to be tested, and f _Carrier Is the fluorescence of the carrier used to dissolve the drug. Generation of dose response plots and IC Using Prism 6 software (GraphPad) ₅₀ The value is obtained.

Example 4 in vivo screening in human myeloma model

1. Single agent screening

Compound ID #1, compound ID #2 and compound ID #3 were tested in the human mm1.s xenograft model for in vivo efficacy studies, as shown in figure 1. Female athymic nude mice were treated with 5.0X 10 ⁶ MM1.S inoculation of human myeloma cells suspended in a mixture of 50% Matrigel (Matrigel) and 50% tissue culture medium in a total volume of 100. mu.L. 18 days after inoculation, the mice were paired into 4 groups of 5 mice each with an average tumor volume of 171mm ³ . Group 1 (G1) was treated with vehicle alone daily for 19 days. Group 2 (G2) was treated daily with Compound ID #1 at 100mg/kg for 19 days. Group 3 (G3) was treated daily with Compound ID #2 at 100mg/kg for 19 days. Group 4 (G4) was treated daily with Compound ID #3 at 100mg/kg for 19 days. Vehicle and compound ID #1, compound ID #2 and compound ID #3 were administered orally by oral gavage. Body weight and tumor measurements were collected twice weekly. The width and length of the tumor were measured in millimeters and converted to tumor volume (cubic millimeters) using the following formula:

compound ID #1 exhibited significantly superior anti-cancer activity when compared to compound ID #2 or compound ID #3 (fig. 1).

2. Combinatorial screening

Compound ID #1, compound ID #2 and compound ID #3 were tested in combination with the FDA approved anticancer drug, Pomalyst (pomalidomide) (fig. 2). Female athymic nude mice were treated with 5.0X 10 ⁶ MM1.S inoculation of human myeloma cells suspended in a mixture of 50% Matrigel (Matrigel) and 50% tissue culture medium in a total volume of 100. mu.L. 18 days after inoculation, the mice were paired into 4 groups of 5 mice each with an average tumor volume of 171mm ³ . Group 1 (G1) was treated with vehicle alone daily for 19 days. Group 2 (G2) was treated daily orally with pomalidomide at 10mg/kg and intraperitoneally with dexamethasone at 0.3mg/kg for 4 days per week until day 19. Group 3 (G3) was treated daily with Compound ID #1 at 100mg/kg for 19 days. Group 4 (G4) was treated daily with Compound ID #2 at 100mg/kg for 19 days. Group 5 (G5) on a daily basisTreated with Compound ID #3 at 100mg/kg for 19 days. Group 6 (G6) was treated daily with Compound ID #1 at 100mg/kg for 19 days, plus pomalidomide/dexamethasone. Group 7 (G7) was treated daily with Compound ID #2 at 100mg/kg for 19 days, plus pomalidomide/dexamethasone. Group 8 (G8) was treated daily with Compound ID #3 at 100mg/kg for 19 days, plus pomalidomide/dexamethasone. Vehicle and compound ID #1, compound ID #2 and compound ID #3 were administered orally by oral gavage. Body weight and tumor measurements were collected twice weekly. The width and length of the tumor were measured in millimeters and converted to tumor volume (cubic millimeters) using the following formula:

compound ID #1 exhibited significantly superior anti-cancer activity when combined with pomalidomide/dexamethasone when compared to compound ID #2 or compound ID #3 in combination with pomalidomide/dexamethasone (figure 2).

Claims (18)

1. A pharmaceutical composition comprising a compound of formula (I):

a pharmaceutically acceptable carrier, and pomalidomide, dexamethasone, or both;

wherein the compound is 4- ((1- (cyclohexylmethyl) -1H-benzo [ d ] imidazol-2-yl) amino) -N-hydroxybenzamide (ID # 1).

2. Use of the pharmaceutical composition of claim 1 in the manufacture of a medicament for treating cancer in a subject.

3. The use of claim 2, wherein the subject is a mammal.

4. The use of claim 2, wherein the subject exhibits a predisposition factor for malignancy selected from the group consisting of: chromosomal translocations associated with malignancy, polyposis, benign monoclonal gammopathy, relativity to people who have or have had cancer or a precancerous condition, and exposure to carcinogens.

5. The use of claim 4, wherein the chromosomal translocation is selected from the group consisting of: philadelphia chromosome and t (14; 18).

6. Use of the pharmaceutical composition of claim 1 in the manufacture of a medicament for slowing the expansion of cancer cells.

7. The use of claim 6, wherein the cell has abnormal growth characterized by hyperplasia, metaplasia, or dysplasia.

8. The use of claim 6, wherein the pharmaceutical composition is administered systemically.

9. The use of claim 6, wherein the pharmaceutical composition is administered topically.

10. The use of claim 6, wherein the use further comprises administering at least one second form of treatment.

11. The use of claim 10, wherein the second therapeutic modality is selected from the group consisting of: radiotherapy, chemotherapy, surgery, immunotherapy, cancer vaccine, radioimmunotherapy, and pharmaceutical compositions comprising an active agent, with the proviso that the active agent is not a compound according to claim 1.

12. The use of claim 10, wherein the second therapeutic modality is selected from the group consisting of: nucleic acid binding components, antiemetic components, hematopoietic colony stimulating factors, anxiolytic agents, and analgesics.

13. The use of claim 10, wherein the second therapeutic form is administered prior to the pharmaceutical composition.

14. The use according to claim 13, wherein the second therapeutic form is administered one or more minutes, hours, days, weeks or months prior to administration of the pharmaceutical composition.

15. The use according to claim 10, wherein the second therapeutic form is administered after administration of the pharmaceutical composition.

16. The use according to claim 15, wherein the second therapeutic form is administered one or more minutes, hours, days, weeks or months after administration of the pharmaceutical composition.

17. The use according to claim 10, wherein the second therapeutic form is administered concurrently with the pharmaceutical composition.

18. The use according to claim 10, wherein the pharmaceutical composition and the second form of treatment are administered cyclically.