COMPOUNDS FOR TREATING PROLIFERATIVE DISEASES

Abstract

The present disclosure provides compounds of Formula (I). The compounds described herein may be useful in treating and/or preventing proliferative diseases (e.g., cancer). Also provided in the present disclosure are pharmaceutical compositions, kits, methods, and uses including or using a compound described herein.

##STR00001##

Claims

1-37. (canceled)

38. A method of treating a proliferative disease in a subject in need thereof, the method comprising administering to the subject an effective amount of a compound of Formula (I): ##STR00062## or a pharmaceutically acceptable salt thereof, wherein: R.sup.1 is independently substituted or unsubstituted alkyl, substituted or unsubstituted carbocyclyl, or substituted or unsubstituted heterocyclyl; R.sup.2 is independently substituted or unsubstituted acyl, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted heterocyclyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, —OR.sup.A, —N(R.sup.B).sub.2, or —SR.sup.A; R.sup.A is substituted or unsubstituted alkyl, substituted or unsubstituted carbocyclyl, substituted or unsubstituted heterocyclyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl or an oxygen protecting group when attached to an oxygen atom or a nitrogen protecting group when attached to a nitrogen atom; each occurrence of R.sup.B is independently substituted or unsubstituted alkyl, substituted or unsubstituted carbocyclyl, substituted or unsubstituted heterocyclyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl; or a nitrogen protecting group; or optionally two instances of R.sup.B are joined to form a substituted or unsubstituted, heterocyclic ring, or substituted or unsubstituted, heteroaryl ring; R.sup.3 is independently hydrogen, substituted or unsubstituted C.sub.1-6 alkyl, or a nitrogen protecting group; and R.sup.4 is independently hydrogen, substituted or unsubstituted C.sub.1-6 alkyl, or a nitrogen protecting group.

39. The method of claim 38, wherein the proliferative disease is cancer.

40. The method of claim 39, wherein the cancer is metastatic cancer.

41. The method of claim 39, wherein the cancer is lung cancer.

42. The method of claim 39, wherein the cancer is prostate cancer.

43. (canceled)

44. The method of claim 38, or a pharmaceutically acceptable salt thereof, wherein R.sup.1 is of the formula: ##STR00063##

45. The method of claim 38, or a pharmaceutically acceptable salt thereof, wherein R.sup.1 is substituted or unsubstituted C.sub.1-6 alkyl.

46. The method of claim 38, or a pharmaceutically acceptable salt thereof, wherein R.sup.1 is of the formula: ##STR00064## wherein: each instance of R.sup.A1 is independently substituted or unsubstituted C.sub.1-6 alkyl; and a is 1, 2, or 3.

47. The method of claim 38, or a pharmaceutically acceptable salt thereof, wherein R.sup.1 is —CH.sub.2CH.sub.2N(CH.sub.3).sub.2 or —CH.sub.2CH.sub.2CH.sub.2N(CH.sub.3).sub.2.

48. The method of claim 38, or a pharmaceutically acceptable salt thereof, wherein R.sup.2 is of the formula: ##STR00065## wherein: R.sup.A2 is independently hydrogen, halogen, or —N(R.sup.C).sub.2; and each instance of R.sup.C is independently substituted or unsubstituted alkyl, or substituted or unsubstituted heterocyclyl.

49. The method of claim 48, or a pharmaceutically acceptable salt thereof, wherein R.sup.A2 is of the formula: ##STR00066##

50. The method of claim 38, or a pharmaceutically acceptable salt thereof, wherein R.sup.2 is —OR.sup.A, and R.sup.A is substituted or unsubstituted carbocyclyl, substituted or unsubstituted heterocyclyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl.

51. The method of claim 50, or a pharmaceutically acceptable salt thereof, wherein R.sup.2 is of the formula: ##STR00067## wherein: R.sup.A3 is —N(R.sup.A4).sub.2, and two instances of R.sup.A4 are joined to form a substituted or unsubstituted, heterocyclic ring, or substituted or unsubstituted, heteroaryl ring.

52. The method of claim 38, or a pharmaceutically acceptable salt thereof, wherein R.sup.2 is —OR.sup.A, and R.sup.A is substituted or unsubstituted phenyl.

53. The method of claim 50, or a pharmaceutically acceptable salt thereof, wherein R.sup.2 is of the formula: ##STR00068## wherein: R.sup.A3 is —N(R.sup.A4).sub.2, and two instances of R.sup.A4 are joined to form a substituted or unsubstituted, heterocyclic ring, or substituted or unsubstituted, heteroaryl ring.

54. The method of claim 38, or a pharmaceutically acceptable salt thereof, wherein R.sup.2 is —SR.sup.A, wherein R.sup.A is substituted or unsubstituted C.sub.1-6 alkyl, or substituted or unsubstituted phenyl.

55. The method of claim 54, or a pharmaceutically acceptable salt thereof, wherein R.sup.2 is —SCH.sub.2CH.sub.2CH.sub.3 or —SPh.

56. The method of claim 38, or a pharmaceutically acceptable salt thereof, wherein R.sup.3 is hydrogen.

57. The method of claim 38, or a pharmaceutically acceptable salt thereof, wherein R.sup.4 is hydrogen.

58. The method of claim 38, wherein the compound is of the formula: ##STR00069## ##STR00070## or a pharmaceutically acceptable salt thereof.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0091] FIG. 1 shows the results of a BCA assay, and of caspase activity in parental (DU145) and high metastatic (DU145LN4) prostate cancer cells upon treatment of compound OBD9. Cells were seeded in 6-well dishes at 50% confluence. Cells were treated with the compound OBD9 after 24 hours. 48 hours later, cells were lysed with lysis buffer and protein was collected and quantified with the BCA assay (Invitrogen). 20 micrograms of protein were loaded on 10% polyacrylamide gels. The gels were blotted onto nitrocellulose and stained with rabbit antibody to PARP-1 and Caspase-3 (both from Cell Signaling) and then visualized with goat, anti-rabbit HRP-conjugated antibody and HRP conjugated GAPDH. FIG. 1 shows that the compound OBD9 induces apoptosis in parental (DU145) and high metastatic (DU145LN4) prostate cancer cells.

[0092] FIG. 2 shows a tubulin polymerization assay for selected compound OBD9. Tubulin polymerization was measured using the tubulin polymerization assay kit from Cytoskeleton Inc., which measures tubulin polymerization kinetic via fluorescent enhancement to the incorporation of a fluorescent reporter into microtubules as polymerization takes place. FIG. 2 shows that OBD9 disrupts (inhibits) tubulin polymerization.

[0093] FIG. 3 shows the effects of two OBD compounds (OBD9 and OBD17) on viability of prostate cancer cell lines (DU145, PC3, and PC3MLN4) and on viability of lung cancer lines (A549, H157, and PC9). 5000 cells were seeded in each well of 96 well plates. 24 hours later, the wells were treated with DMSO or the OBD compounds (OBD9 and OBD17) at the respective indicated concentrations (0.013 μM, 0.04 μM, 0.12 μM, 0.37 μM, 1.1 μM, 3.3 μM, or 10 μM). 72 hours after treatment, the cells were analyzed using the MTT cell viability assay (Sigma).

[0094] FIG. 4 shows imaging of microtubule stability in in PC3M-prostate cancer cells. Cells were seeded on glass coverslips. 24 hours later, cells were treated with compound OBD9. After the indicated times (24 hours, 48 hours, and 72 hours), cells were fixed with paraformaldehyde (4% in PBS), permeabilized Triton-X100 (0.02% in Phosphate buffered saline), then stained with mouse antibody to human tubulin (1:200 dilution) and then with 1:400 goat anti-mouse IgC (Invitrogen) and viewed via confocal microscopy at (63× objective). FIG. 4 shows that OBD9 interferes with microtubule (tubulin) stability in PC3M-prostate cancer cells.

DETAILED DESCRIPTION OF CERTAIN EMBODIMENTS OF THE INVENTION

[0095] The present disclosure provides compounds of Formula (I). The compounds described herein are useful in treating and/or preventing proliferative diseases (e.g., cancer). The compounds may act by inhibiting growth of cells (e.g., aggressive cancer cells). Also provided in the present disclosure are pharmaceutical compositions, kits, methods, and uses including a compound described herein.

Compounds

[0096] One aspect of the present disclosure relates to compounds of Formula (I) as described herein. The compounds described herein are compounds that are useful in treating and/or preventing proliferative diseases in a subject. The compounds may also be useful for contacting a biological sample, tissue, or cell with an effective amount of a compound or pharmaceutical composition described herein. In certain embodiments, a compound described herein is a compound of Formula (I), or a pharmaceutically acceptable salt, solvate, hydrate, polymorph, co-crystal, tautomer, stereoisomer, isotopically labeled derivative, or prodrug thereof. In certain embodiments, a compound described herein is a compound of Formula (I), or a pharmaceutically acceptable salt thereof.

[0097] In certain embodiments, a compound described herein is of Formula (I):

##STR00010##

or a pharmaceutically acceptable salt, solvate, hydrate, polymorph, co-crystal, tautomer, stereoisomer, isotopically labeled derivative, or prodrug thereof, wherein:

[0098] R.sup.1 is independently substituted or unsubstituted alkyl, substituted or unsubstituted carbocyclyl, or substituted or unsubstituted heterocyclyl;

[0099] R.sup.2 is independently substituted or unsubstituted acyl, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted heterocyclyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, —OR.sup.A, —N(R.sup.B).sub.2, or —SR.sup.A;

[0100] R.sup.A is substituted or unsubstituted alkyl, substituted or unsubstituted carbocyclyl, substituted or unsubstituted heterocyclyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl or an oxygen protecting group when attached to an oxygen atom or a nitrogen protecting group when attached to a nitrogen atom;

[0101] each occurrence of R.sup.B is independently substituted or unsubstituted alkyl, substituted or unsubstituted carbocyclyl, substituted or unsubstituted heterocyclyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl; or a nitrogen protecting group; or optionally two instances of R.sup.B are joined to form a substituted or unsubstituted, heterocyclic ring, or substituted or unsubstituted, heteroaryl ring;

[0102] R.sup.3 is independently hydrogen, substituted or unsubstituted C.sub.1-6 alkyl, or a nitrogen protecting group; and

[0103] R.sup.4 is independently hydrogen, substituted or unsubstituted C.sub.1-6 alkyl, or a nitrogen protecting group.

[0104] Formula (I) includes substituent R.sup.1. In certain embodiments, R.sup.1 is substituted or unsubstituted alkyl (e.g., substituted or unsubstituted C.sub.1-6 alkyl). In certain embodiments, R.sup.1 is independently —CH.sub.3, —CH.sub.2CH.sub.3, —CH.sub.2CH.sub.2CH.sub.3, —CH.sub.2CH.sub.2CH.sub.2CH.sub.3. In certain embodiments, R.sup.1 is substituted alkyl, e.g., heterocyclalkyl. In certain embodiments, R.sup.1 is of the formula:

##STR00011##

In certain embodiments, R is of the formula:

##STR00012##

In certain embodiments, R.sup.1 is substituted or unsubstituted carbocyclyl (e.g., substituted or unsubstituted, 3- to 7-membered, monocyclic carbocyclyl comprising zero, one, or two double bonds in the carbocyclic ring system). In certain embodiments, R.sup.1 is substituted or unsubstituted heterocyclyl (e.g., substituted or unsubstituted, 3- to 9-membered, monocyclic heterocyclyl comprising zero, one, or two double bonds in the heterocyclic ring system, wherein one, two, or three atoms in the heterocyclic ring system are independently nitrogen, oxygen, or sulfur). In certain embodiments, R.sup.1 is of the formula

##STR00013##

In certain embodiments, R.sup.1 is of the formula:

##STR00014##

In certain embodiments, R.sup.1 is —(CH.sub.2).sub.aN(R.sup.A1).sub.2, a is 1, 2, 3, 4, 5, or 6, and each occurrence of R.sup.A1 is independently hydrogen, or substituted or unsubstituted C.sub.1-6 alkyl (e.g., R.sup.1 is —(CH.sub.2).sub.2NMe.sub.2)). In certain embodiments, R.sup.1 is —(CH.sub.2).sub.3NMe.sub.2). In certain embodiments, R.sup.A1 is independently —CH.sub.2CH.sub.3, or —CH.sub.2CH.sub.2CH.sub.3.

[0105] Formula (I) includes substituent R.sup.2. In certain embodiments, R.sup.2 is substituted or unsubstituted acyl (e.g., acetyl). In certain embodiments, R.sup.2 is of the formula:

##STR00015##

In certain embodiments, R.sup.A2 is independently hydrogen, halogen (e.g., F, Cl, Br, or I), or —N(R.sup.C). In certain embodiments, R.sup.A2 is of the formula:

##STR00016##

In certain embodiments, R.sup.A2 is of the formula:

##STR00017##

In certain embodiments, R.sup.C is independently substituted or unsubstituted alkyl, or substituted or unsubstituted heterocyclyl. When Formula (I) includes two or more instances of the substituent R.sup.C, any two instances of R.sup.C may be the same or different from each other. In certain embodiments, at least one instance of R.sup.C is substituted or unsubstituted C.sub.1-6 alkyl (e.g., -Me). In certain embodiments, at least one instance of R.sup.C is substituted or unsubstituted heterocyclyl (e.g., substituted or unsubstituted, 3- to 9-membered, monocyclic heterocyclyl comprising zero, one, or two double bonds in the heterocyclic ring system, wherein one, two, or three atoms in the heterocyclic ring system are independently nitrogen, oxygen, or sulfur).

[0106] In certain embodiments, R.sup.2 is —OR.sup.A. In certain embodiments, R.sup.A is substituted or unsubstituted carbocyclyl (e.g., substituted or unsubstituted, 3- to 7-membered, monocyclic carbocyclyl comprising zero, one, or two double bonds in the carbocyclic ring system). In certain embodiments, R.sup.A is substituted or unsubstituted heterocyclyl (e.g., substituted or unsubstituted, 3- to 9-membered, monocyclic heterocyclyl comprising zero, one, or two double bonds in the heterocyclic ring system, wherein one, two, or three atoms in the heterocyclic ring system are independently nitrogen, oxygen, or sulfur). In certain embodiments, RA is a substituted or unsubstituted, aryl ring (e.g., substituted or unsubstituted, 6- to 10-membered aryl). In certain embodiments, R.sup.2 is —OR.sup.A (e.g. —O(substituted or unsubstituted phenyl) (e.g., —OPh)).

In certain embodiments, R.sup.2 is of the formula:

##STR00018##

wherein R.sup.A3 is —N(R.sup.A4). In certain embodiments, R.sup.A4 is independently substituted or unsubstituted alkyl, substituted or unsubstituted carbocyclyl, substituted or unsubstituted heterocyclyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl. In certain embodiments. R.sup.2 is of the formula:

##STR00019##

wherein R.sup.A3 is —N(R.sup.A4).sub.2. In certain embodiments, two instances of R.sup.A4 are joined to form a substituted or unsubstituted, heterocyclic ring (e.g., substituted or unsubstituted, 5- to 6-membered, monocyclic heterocyclic ring comprising zero, one, or two double bonds in the heterocyclic ring system, wherein one, two, or three atoms in the heterocyclic ring system are independently nitrogen, oxygen, or sulfur). In certain embodiments, two instances of R.sup.A4 are joined to form a substituted or unsubstituted heteroaryl ring (e.g., substituted or unsubstituted, 5- to 6-membered, monocyclic heteroaryl ring, wherein one, two, three, or four atoms in the heteroaryl ring system are independently nitrogen, oxygen, or sulfur). In certain embodiments, R.sup.A3 is of the formula:

##STR00020##

[0107] In certain embodiments, R.sup.2 is —SR.sup.A. In certain embodiments, RA is substituted or unsubstituted C.sub.1-6 alkyl. In certain embodiments, R.sup.A is substituted or unsubstituted benzyl, or substituted or unsubstituted phenyl.

[0108] Formula (I) includes substituent R.sup.3. In certain embodiments, R.sup.3 is H. In certain embodiments, R.sup.3 is substituted or unsubstituted alkyl (e.g., substituted or unsubstituted C.sub.1-6 alkyl). In certain embodiments. R.sup.3 is a nitrogen protecting group (e.g., benzyl (Bn), t-butyl carbonate (BOC or Boc), benzyl carbamate (Cbz), 9-fluorenylmethyl carbonate (Fmoc), trifluoroacetyl, triphenylmethyl, acetyl, or p-toluenesulfonamide (Ts)).

[0109] Formula (I) includes substituent R. In certain embodiments, R.sup.4 is H. In certain embodiments, R.sup.4 is substituted or unsubstituted alkyl (e.g., substituted or unsubstituted C.sub.1-6 alkyl). In certain embodiments, R.sup.4 is a nitrogen protecting group (e.g., benzyl (Bn), t-butyl carbonate (BOC or Boc), benzyl carbamate (Cbz), 9-fluorenylmethyl carbonate (Fmoc), trifluoroacetyl, triphenylmethyl, acetyl, or p-toluenesulfonamide (Ts)).

[0110] In certain embodiments, the compound of Formula (I) is of the following formula:

##STR00021##

or a pharmaceutically acceptable salt thereof.

[0111] In certain embodiments, the compound of Formula (I) is of one of the following formulae:

##STR00022##

or a pharmaceutically acceptable salt thereof.

[0112] In certain embodiments, the compound of Formula (I) is of one of the following formulae:

##STR00023##

or a pharmaceutically acceptable salt thereof.

[0113] In certain embodiments, the compound of Formula (I) is of one of the following formulae:

##STR00024##

or a pharmaceutically acceptable salt thereof.

[0114] In certain embodiments, the compound of Formula (I) is of the formula:

##STR00025## ##STR00026##

or a pharmaceutically acceptable salt thereof.

Pharmaceutical Compositions, Kits, and Administration

[0115] The present disclosure provides pharmaceutical compositions comprising a compound described herein, or a pharmaceutically acceptable salt thereof, and optionally a pharmaceutically acceptable excipient. In certain embodiments, a pharmaceutical composition described herein comprises a compound described herein, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient. The pharmaceutical compositions described herein are useful in treating and/or preventing proliferative diseases (e.g., cancer (e.g., lung cancer, breast cancer, leukemia, lymphoma, melanoma, multiple myeloma, Ewing's sarcoma, osteosarcoma, brain cancer, neuroblastoma), benign neoplasms, angiogenesis, inflammatory diseases, autoinflammatory diseases, and autoimmune diseases) in a subject, biological sample, tissue, or cell.

[0116] In certain embodiments, a subject being treated herein is an animal. The animal may be of either sex and may be at any stage of development. In certain embodiments, the subject is a human. In certain embodiments, the subject is a non-human animal. In certain embodiments, the subject is a mammal (e.g., non-human mammal). In certain embodiments, the subject is a domesticated animal, such as a dog, cat, cow, pig, horse, sheep, or goat. In certain embodiments, a subject being treated herein is a companion animal such as a dog or cat. In certain embodiments, a subject being treated herein is a livestock animal such as a cow, pig, horse, sheep, or goat. In certain embodiments, a subject being treated herein is a zoo animal. In another embodiment, a subject being treated herein is a research animal such as a rodent (e.g., mouse, rat), dog, pig, or non-human primate. In certain embodiments, the animal is a genetically engineered animal. In certain embodiments, the animal is a transgenic or genetically engineered animal (e.g., transgenic mice and transgenic pigs). In certain embodiments, a subject described herein is a fish or reptile.

[0117] In certain embodiments, the cell contacted with an effective amount of a compound or pharmaceutical composition described herein is in vitro. In certain embodiments, the contacted cell is ex vivo. In certain embodiments, the cell described herein is in vivo. In certain embodiments, the cell described herein is a malignant cell (e.g., malignant blood cell).

[0118] In certain embodiments, the compound described herein is provided in an effective amount in the pharmaceutical composition. In certain embodiments, the effective amount is a therapeutically effective amount (e.g., amount effective for treating a proliferative disease in a subject in need thereof). In certain embodiments, the proliferative disease is cancer. In certain embodiments, the proliferative disease is metastatic cancer. In certain embodiments, the effective amount is a prophylactically effective amount (e.g., amount effective for preventing a proliferative disease in a subject in need thereof and/or for keeping a subject in need thereof in remission of a proliferative disease).

[0119] Pharmaceutical compositions described herein can be prepared by any method known in the art of pharmacology. In general, such preparatory methods include bringing the compound described herein (i.e., the “active ingredient”) into association with a carrier or excipient, and/or one or more other accessory ingredients, and then, if necessary and/or desirable, shaping, and/or packaging the product into a desired single- or multi-dose unit.

[0120] Pharmaceutical compositions can be prepared, packaged, and/or sold in bulk, as a single unit dose, and/or as a plurality of single unit doses. A “unit dose” is a discrete amount of the pharmaceutical composition comprising a predetermined amount of the active ingredient. The amount of the active ingredient is generally equal to the dosage of the active ingredient which would be administered to a subject and/or a convenient fraction of such a dosage, such as one-half or one-third of such a dosage.

[0121] Relative amounts of the active ingredient, the pharmaceutically acceptable excipient, and/or any additional ingredients in a pharmaceutical composition described herein will vary, depending upon the identity, size, and/or condition of the subject treated and further depending upon the route by which the composition is to be administered. The composition may comprise between 0.1% and 100% (w/w) active ingredient.

[0122] Pharmaceutically acceptable excipients used in the manufacture of provided pharmaceutical compositions include inert diluents, dispersing and/or granulating agents, surface active agents and/or emulsifiers, disintegrating agents, binding agents, preservatives, buffering agents, lubricating agents, and/or oils. Excipients such as cocoa butter and suppository waxes, coloring agents, coating agents, sweetening, flavoring, and perfuming agents may also be present in the composition.

[0123] Liquid dosage forms for oral and parenteral administration include pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups and elixirs. In addition to the active ingredients, the liquid dosage forms may comprise inert diluents commonly used in the art such as, for example, water or other solvents, solubilizing agents and emulsifiers such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, dimethylformamide, oils (e.g., cottonseed, groundnut, corn, germ, olive, castor, and sesame oils), glycerol, tetrahydrofurfuryl alcohol, polyethylene glycols and fatty acid esters of sorbitan, and mixtures thereof. Besides inert diluents, the oral compositions can include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents. In certain embodiments for parenteral administration, the conjugates described herein are mixed with solubilizing agents such as Cremophor®, alcohols, oils, modified oils, glycols, polysorbates, cyclodextrins, polymers, and mixtures thereof.

[0124] Injectable preparations, for example, sterile injectable aqueous or oleaginous suspensions can be formulated according to the known art using suitable dispersing or wetting agents and suspending agents. The sterile injectable preparation can be a sterile injectable solution, suspension, or emulsion in a nontoxic parenterally acceptable diluent or solvent, for example, as a solution in 1,3-butanediol. Among the acceptable vehicles and solvents that can be employed are water, Ringer's solution, U.S.P., and isotonic sodium chloride solution. In addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium. For this purpose any bland fixed oil can be employed including synthetic mono- or di-glycerides. In addition, fatty acids such as oleic acid are used in the preparation of injectables.

[0125] The injectable formulations can be sterilized, for example, by filtration through a bacterial-retaining filter, or by incorporating sterilizing agents in the form of sterile solid compositions which can be dissolved or dispersed in sterile water or other sterile injectable medium prior to use.

[0126] In order to prolong the effect of a drug, it is often desirable to slow the absorption of the drug from subcutaneous or intramuscular injection. This can be accomplished by the use of a liquid suspension of crystalline or amorphous material with poor water solubility. The rate of absorption of the drug then depends upon its rate of dissolution, which, in turn, may depend upon crystal size and crystalline form. Alternatively, delayed absorption of a parenterally administered drug form may be accomplished by dissolving or suspending the drug in an oil vehicle.

[0127] Compositions for rectal or vaginal administration are typically suppositories which can be prepared by mixing the conjugates described herein with suitable non-irritating excipients or carriers such as cocoa butter, polyethylene glycol, or a suppository wax which are solid at ambient temperature but liquid at body temperature and therefore melt in the rectum or vaginal cavity and release the active ingredient.

[0128] Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In such solid dosage forms, the active ingredient is mixed with at least one inert, pharmaceutically acceptable excipient or carrier such as sodium citrate or dicalcium phosphate and/or (a) fillers or extenders such as starches, lactose, sucrose, glucose, mannitol, and silicic acid, (b) binders such as, for example, carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidinone, sucrose, and acacia, (c) humectants such as glycerol, (d) disintegrating agents such as agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates, and sodium carbonate, (e) solution retarding agents such as paraffin, (f) absorption accelerators such as quaternary ammonium compounds, (g) wetting agents such as, for example, cetyl alcohol and glycerol monostearate, (h) absorbents such as kaolin and bentonite clay, and (i) lubricants such as talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, and mixtures thereof. In the case of capsules, tablets, and pills, the dosage form may include a buffering agent.

[0129] Solid compositions of a similar type can be employed as fillers in soft and hard-filled gelatin capsules using such excipients as lactose or milk sugar as well as high molecular weight polyethylene glycols and the like. The solid dosage forms of tablets, dragees, capsules, pills, and granules can be prepared with coatings and shells such as enteric coatings and other coatings well known in the art of pharmacology. They may optionally comprise opacifying agents and can be of a composition that they release the active ingredient(s) only, or preferentially, in a certain part of the intestinal tract, optionally, in a delayed manner. Examples of encapsulating compositions which can be used include polymeric substances and waxes. Solid compositions of a similar type can be employed as fillers in soft and hard-filled gelatin capsules using such excipients as lactose or milk sugar as well as high molecular weight polyethylene glycols and the like.

[0130] The active ingredient can be in a micro-encapsulated form with one or more excipients as noted above. The solid dosage forms of tablets, dragees, capsules, pills, and granules can be prepared with coatings and shells such as enteric coatings, release controlling coatings, and other coatings well known in the pharmaceutical formulating art. In such solid dosage forms the active ingredient can be admixed with at least one inert diluent such as sucrose, lactose, or starch. Such dosage forms may comprise, as is normal practice, additional substances other than inert diluents, e.g., tableting lubricants and other tableting aids such a magnesium stearate and microcrystalline cellulose. In the case of capsules, tablets and pills, the dosage forms may comprise buffering agents. They may optionally comprise opacifying agents and can be of a composition that they release the active ingredient(s) only, or preferentially, in a certain part of the intestinal tract, optionally, in a delayed manner. Examples of encapsulating agents which can be used include polymeric substances and waxes.

[0131] Dosage forms for topical and/or transdermal administration of a compound described herein may include ointments, pastes, creams, lotions, gels, powders, solutions, sprays, inhalants, and/or patches. Generally, the active ingredient is admixed under sterile conditions with a pharmaceutically acceptable carrier or excipient and/or any needed preservatives and/or buffers as can be required. Additionally, the present disclosure contemplates the use of transdermal patches, which often have the added advantage of providing controlled delivery of an active ingredient to the body. Such dosage forms can be prepared, for example, by dissolving and/or dispensing the active ingredient in the proper medium. Alternatively or additionally, the rate can be controlled by either providing a rate controlling membrane and/or by dispersing the active ingredient in a polymer matrix and/or gel.

[0132] Suitable devices for use in delivering intradermal pharmaceutical compositions described herein include short needle devices. Intradermal compositions can be administered by devices which limit the effective penetration length of a needle into the skin. Alternatively or additionally, conventional syringes can be used in the classical mantoux method of intradermal administration. Jet injection devices which deliver liquid formulations to the dermis via a liquid jet injector and/or via a needle which pierces the stratum corneum and produces a jet which reaches the dermis are suitable. Ballistic powder/particle delivery devices which use compressed gas to accelerate the compound in powder form through the outer layers of the skin to the dermis are suitable.

[0133] Formulations suitable for topical administration include, but are not limited to, liquid and/or semi-liquid preparations such as liniments, lotions, oil-in-water and/or water-in-oil emulsions such as creams, ointments, and/or pastes, and/or solutions and/or suspensions. Topically administrable formulations may, for example, comprise from about 1% to about 10% (w/w) active ingredient, although the concentration of the active ingredient can be as high as the solubility limit of the active ingredient in the solvent. Formulations for topical administration may further comprise one or more of the additional ingredients described herein.

[0134] A pharmaceutical composition described herein can be prepared, packaged, and/or sold in a formulation suitable for pulmonary administration via the buccal cavity. Such a formulation may comprise dry particles which comprise the active ingredient and which have a diameter in the range from about 0.5 to about 7 nanometers, or from about 1 to about 6 nanometers. Such compositions are conveniently in the form of dry powders for administration using a device comprising a dry powder reservoir to which a stream of propellant can be directed to disperse the powder and/or using a self-propelling solvent/powder dispensing container such as a device comprising the active ingredient dissolved and/or suspended in a low-boiling propellant in a sealed container. Such powders comprise particles wherein at least 98% of the particles by weight have a diameter greater than 0.5 nanometers and at least 95% of the particles by number have a diameter less than 7 nanometers. Alternatively, at least 95% of the particles by weight have a diameter greater than 1 nanometer and at least 90% of the particles by number have a diameter less than 6 nanometers. Dry powder compositions may include a solid fine powder diluent such as sugar and am conveniently provided in a unit dose form.

[0135] Low boiling propellants generally include liquid propellants having a boiling point of below 65° F. at atmospheric pressure. Generally the propellant may constitute 50 to 99.9% b (w/w) of the composition, and the active ingredient may constitute 0.1 to 20% (w/w) of the composition. The propellant may further comprise additional ingredients such as a liquid non-ionic and/or solid anionic surfactant and/or a solid diluent (which may have a particle size of the same order as particles comprising the active ingredient).

[0136] Pharmaceutical compositions described herein formulated for pulmonary delivery may provide the active ingredient in the form of droplets of a solution and/or suspension. Such formulations can be prepared, packaged, and/or sold as aqueous and/or dilute alcoholic solutions and/or suspensions, optionally sterile, comprising the active ingredient, and may conveniently be administered using any nebulization and/or atomization device. Such formulations may further comprise one or more additional ingredients including, but not limited to, a flavoring agent such as saccharin sodium, a volatile oil, a buffering agent, a surface active agent, and/or a preservative such as methylhydroxybenzoate. The droplets provided by this route of administration may have an average diameter in the range from about 0.1 to about 200 nanometers.

[0137] Formulations described herein as being useful for pulmonary delivery are useful for intranasal delivery of a pharmaceutical composition described herein. Another formulation suitable for intranasal administration is a coarse powder comprising the active ingredient and having an average particle from about 0.2 to 500 micrometers. Such a formulation is administered by rapid inhalation through the nasal passage from a container of the powder held close to the nares.

[0138] Formulations for nasal administration may, for example, comprise from about as little as 0.1% (w/w) to as much as 100% (w/w) of the active ingredient, and may comprise one or more of the additional ingredients described herein. A pharmaceutical composition described herein can be prepared, packaged, and/or sold in a formulation for buccal administration. Such formulations may, for example, be in the form of tablets and/or lozenges made using conventional methods, and may contain, for example, 0.1 to 20% (w/w) active ingredient, the balance comprising an orally dissolvable and/or degradable composition and, optionally, one or more of the additional ingredients described herein. Alternately, formulations for buccal administration may comprise a powder and/or an aerosolized and/or atomized solution and/or suspension comprising the active ingredient. Such powdered, aerosolized, and/or aerosolized formulations, when dispersed, may have an average particle and/or droplet size in the range from about 0.1 to about 200 nanometers, and may further comprise one or more of the additional ingredients described herein.

[0139] A pharmaceutical composition described herein can be prepared, packaged, and/or sold in a formulation for ophthalmic administration. Such formulations may, for example, be in the form of eye drops including, for example, a 0.1-1.0% (w/w) solution and/or suspension of the active ingredient in an aqueous or oily liquid carrier or excipient. Such drops may further comprise buffering agents, salts, and/or one or more other of the additional ingredients described herein. Other ophthalmically-administrable formulations which are useful include those which comprise the active ingredient in microcrystalline form and/or in a liposomal preparation. Ear drops and/or eye drops are also contemplated as being within the scope of this disclosure.

[0140] Although the descriptions of pharmaceutical compositions provided herein are principally directed to pharmaceutical compositions which are suitable for administration to humans, such compositions are generally suitable for administration to animals of all sorts. Modification of pharmaceutical compositions suitable for administration to humans in order to render the compositions suitable for administration to various animals is well understood, and the ordinarily skilled veterinary pharmacologist can design and/or perform such modification with ordinary experimentation.

[0141] The compounds provided herein are typically formulated in dosage unit form for ease of administration and uniformity of dosage. It will be understood, however, that the total daily usage of the compositions described herein will be decided by a physician within the scope of sound medical judgment. The specific therapeutically effective dose level for any particular subject or organism will depend upon a variety of factors including the disease being treated and the severity of the disorder; the activity of the specific active ingredient employed, the specific composition employed; the age, body weight, general health, sex, and diet of the subject; the time of administration, route of administration, and rate of excretion of the specific active ingredient employed; the duration of the treatment; drugs used in combination or coincidental with the specific active ingredient employed; and like factors well known in the medical arts.

[0142] The compounds and compositions provided herein can be administered by any route, including enteral (e.g., oral), parenteral, intravenous, intramuscular, intra-arterial, intramedullary, intrathecal, subcutaneous, intraventricular, transdermal, intradermal, rectal, intravaginal, intraperitoneal, topical (as by powders, ointments, creams, and/or drops), mucosal, nasal, bucal, sublingual; by intratracheal instillation, bronchial instillation, and/or inhalation; and/or as an oral spray, nasal spray, and/or aerosol. Specifically contemplated routes are oral administration, intravenous administration (e.g., systemic intravenous injection), regional administration via blood and/or lymph supply, and/or direct administration to an affected site. In general, the most appropriate route of administration will depend upon a variety of factors including the nature of the agent (e.g., its stability in the environment of the gastrointestinal tract), and/or the condition of the subject (e.g., whether the subject is able to tolerate oral administration). In certain embodiments, the compound or pharmaceutical composition described herein is suitable for topical administration to the eye of a subject.

[0143] The exact amount of a compound required to achieve an effective amount will vary from subject to subject, depending, for example, on species, age, and general condition of a subject, severity of the side effects or disorder, identity of the particular compound, mode of administration, and the like. An effective amount may be included in a single dose (e.g., single oral dose) or multiple doses (e.g., multiple oral doses). In certain embodiments, when multiple doses are administered to a subject or applied to a biological sample, tissue, or cell, any two doses of the multiple doses include different or substantially the same amounts of a compound described herein. In certain embodiments, when multiple doses are administered to a subject or applied to a biological sample, tissue, or cell, the frequency of administering the multiple doses to the subject or applying the multiple doses to the tissue or cell is three doses a day, two doses a day, one dose a day, one dose every other day, one dose every third day, one dose every week, one dose every two weeks, one dose every three weeks, or one dose every four weeks. In certain embodiments, the frequency of administering the multiple doses to the subject or applying the multiple doses to the tissue or cell is one dose per day. In certain embodiments, the frequency of administering the multiple doses to the subject or applying the multiple doses to the tissue or cell is two doses per day. In certain embodiments, the frequency of administering the multiple doses to the subject or applying the multiple doses to the tissue or cell is three doses per day. In certain embodiments, when multiple doses are administered to a subject or applied to a biological sample, tissue, or cell, the duration between the first dose and last dose of the multiple doses is one day, two days, four days, one week, two weeks, three weeks, one month, two months, three months, four months, six months, nine months, one year, two years, three years, four years, five years, seven years, ten years, fifteen years, twenty years, or the lifetime of the subject, biological sample, tissue, or cell. In certain embodiments, the duration between the first dose and last dose of the multiple doses is three months, six months, or one year. In certain embodiments, the duration between the first dose and last dose of the multiple doses is the lifetime of the subject, biological sample, tissue, or cell. In certain embodiments, a dose (e.g., a single dose, or any dose of multiple doses) described herein includes independently between 0.1 μg and 1 μg, between 0.001 mg and 0.01 mg, between 0.01 mg and 0.1 mg, between 0.1 mg and 1 mg, between 1 mg and 3 mg, between 3 mg and 10 mg, between 10 mg and 30 mg, between 30 mg and 100 mg, between 100 mg and 300 mg, between 300 mg and 1,000 mg, or between 1 g and 10 g. inclusive, of a compound described herein. In certain embodiments, a dose described herein includes independently between 1 mg and 3 mg, inclusive, of a compound described herein. In certain embodiments, a dose described herein includes independently between 3 mg and 10 mg, inclusive, of a compound described herein. In certain embodiments, a dose described herein includes independently between 10 mg and 30 mg, inclusive, of a compound described herein. In certain embodiments, a dose described herein includes independently between 30 mg and 100 mg, inclusive, of a compound described herein.

[0144] Dose ranges as described herein provide guidance for the administration of provided pharmaceutical compositions to an adult. The amount to be administered to, for example, a child or an adolescent can be determined by a medical practitioner or person skilled in the art and can be lower or the same as that administered to an adult.

[0145] A compound or composition, as described herein, can be administered in combination with one or more additional pharmaceutical agents (e.g., therapeutically and/or prophylactically active agents) useful in treating and/or preventing a proliferative disease. The compounds or compositions can be administered in combination with additional pharmaceutical agents that improve their activity (e.g., activity (e.g., potency and/or efficacy) in treating a proliferative disease in a subject in need thereof, and/or in preventing a proliferative disease in a subject in need thereof), improve bioavailability, improve safety, reduce drug resistance, reduce and/or modify metabolism, inhibit excretion, and/or modify distribution in a subject, biological sample, tissue, or cell. It will also be appreciated that the therapy employed may achieve a desired effect for the same disorder, and/or it may achieve different effects. In certain embodiments, a pharmaceutical composition described herein including a compound described herein and an additional pharmaceutical agent shows a synergistic effect that is absent in a pharmaceutical composition including one of the compound and the additional pharmaceutical agent, but not both.

[0146] The compound or composition can be administered concurrently with, prior to, or subsequent to one or more additional pharmaceutical agents, which may be useful as, e.g., combination therapies in treating and/or preventing a proliferative disease. Pharmaceutical agents include therapeutically active agents. Pharmaceutical agents also include prophylactically active agents. Pharmaceutical agents include small organic molecules such as drug compounds (e.g., compounds approved for human or veterinary use by the U.S. Food and Drug Administration as provided in the Code of Federal Regulations (CFR)), peptides, proteins, carbohydrates, monosaccharides, oligosaccharides, polysaccharides, nucleoproteins, mucoproteins, lipoproteins, synthetic polypeptides or proteins, small molecules linked to proteins, glycoproteins, steroids, nucleic acids, DNAs, RNAs, nucleotides, nucleosides, oligonucleotides, antisense oligonucleotides, lipids, hormones, vitamins, and cells. In certain embodiments, the additional pharmaceutical agent is a pharmaceutical agent useful in treating a proliferative disease. In certain embodiments, the additional pharmaceutical agent is a pharmaceutical agent useful in preventing a proliferative disease. In certain embodiments, the additional pharmaceutical agent is a pharmaceutical agent approved by a regulatory agency (e.g., the US FDA) for treating and/or preventing a proliferative disease. Each additional pharmaceutical agent may be administered at a dose and/or on a time schedule determined for that pharmaceutical agent. The additional pharmaceutical agents may also be administered together with each other and/or with the compound or composition described herein in a single dose or administered separately in different doses. The particular combination to employ in a regimen will take into account compatibility of the compound described herein with the additional pharmaceutical agent(s) and/or the desired therapeutic and/or prophylactic effect to be achieved. In general, it is expected that the additional pharmaceutical agent(s) in combination be utilized at levels that do not exceed the levels at which they are utilized individually. In some embodiments, the levels utilized in combination will be lower than those utilized individually.

[0147] In certain embodiments, the additional pharmaceutical agent is an anti-proliferative agent (e.g., anti-cancer agent). In certain embodiments, the additional pharmaceutical agent is an anti-angiogenesis agent, anti-inflammatory agent, immunosuppressant, anti-bacterial agent, anti-viral agent, cardiovascular agent, cholesterol-lowering agent, anti-diabetic agent, anti-allergic agent, pain-relieving agent, or a combination thereof. In certain embodiments, the compounds described herein or pharmaceutical compositions can be administered in combination with an anti-cancer therapy including, but not limited to, transplantation (e.g., bone marrow transplantation, stem cell transplantation), surgery, radiation therapy, immunotherapy, and chemotherapy.

[0148] Also encompassed by the disclosure are kits (e.g., pharmaceutical packs). The kits provided may comprise a pharmaceutical composition or compound described herein and a container (e.g., a vial, ampule, bottle, syringe, and/or dispenser package, or other suitable container). In some embodiments, provided kits may optionally further include a second container comprising a pharmaceutical excipient for dilution or suspension of a pharmaceutical composition or compound described herein. In some embodiments, the pharmaceutical composition or compound described herein provided in the first container and the second container are combined to form one unit dosage form.

[0149] In certain embodiments, a kit described herein includes a first container comprising a compound or pharmaceutical composition described herein. In certain embodiments, a kit described herein is useful in treating a proliferative disease (e.g., metastatic cancer) in a subject in need thereof, and/or preventing a proliferative disease in a subject in need thereof.

[0150] In certain embodiments, a kit described herein further includes instructions for using the compound or pharmaceutical composition included in the kit. A kit described herein may also include information as required by a regulatory agency such as the U.S. Food and Drug Administration (FDA). In certain embodiments, the information included in the kits is prescribing information. In certain embodiments, the kits and instructions provide for treating a proliferative disease in a subject in need thereof, and/or preventing a proliferative disease in a subject in need thereof. A kit described herein may include one or more additional pharmaceutical agents described herein as a separate composition.

Methods of Treatment

[0151] The compounds described herein may: [0152] exhibit cytotoxic or growth inhibitory effect on cancer cell lines maintained in vitro or in animal studies using a scientifically acceptable cancer cell xenograft model; [0153] exhibit a therapeutic profile (e.g., optimum safety and curative effect) that is superior to existing chemotherapeutic agents; and/or [0154] exhibit a pharmacokinetic profile (e.g., increased solubility and increased bioavailability) that is superior to existing chemotherapeutic agents.

[0155] The present disclosure provides methods of treating a proliferative disease in a subject in need thereof, the methods comprising administering to the subject an effective amount (e.g., therapeutically effective amount) of a compound, or pharmaceutical composition thereof, described herein.

[0156] Another aspect of the present disclosure relates to methods of preventing proliferative disease in a subject in need thereof, the methods comprising administering to the subject an effective amount (e.g., prophylactically effective amount) of a compound, or pharmaceutical composition thereof, described herein.

[0157] The compounds and pharmaceutical compositions described herein are useful in treating and/or preventing proliferative diseases. In certain embodiments, the proliferative disease is cancer. In certain embodiments, the proliferative disease is metastatic cancer. In certain embodiments, the proliferative disease is an inflammatory disease. In certain embodiments, the proliferative disease is an autoimmune disease.

[0158] In certain embodiments, the method described herein further includes administering to the subject an additional pharmaceutical agent. In certain embodiments, the method described herein further includes contacting the biological sample with an additional pharmaceutical agent. In certain embodiments, the method described herein further includes contacting the tissue with an additional pharmaceutical agent. In certain embodiments, the method described herein further includes contacting the cell with an additional pharmaceutical agent. In certain embodiments, the method described herein further includes radiotherapy, immunotherapy, and/or transplantation (e.g., bone marrow transplantation).

Uses

[0159] In another aspect, the present disclosure provides the compounds described herein for a method of treating a proliferative disease in the disclosure.

[0160] In still another aspect, the present disclosure provides the pharmaceutical compositions described herein for use in a method of treating a proliferative disease in the disclosure.

[0161] In still another aspect, the present disclosure provides uses of the compounds described herein in a method of treating a proliferative disease in the disclosure.

[0162] In further another aspect, the present disclosure provides uses of the pharmaceutical compositions described herein in a method of treating a proliferative disease in the disclosure.

EXAMPLES

[0163] In order that the present disclosure may be more fully understood, the following examples are set forth. The synthetic and biological examples described in this application are offered to illustrate the compounds, pharmaceutical compositions, and methods provided herein and are not to be construed in any way as limiting their scope.

Example 1A. Selected Compounds and Biological Activity

[0164] Table 1 below shows the results of a viability assay: the effect of selected exemplary compounds at concentrations of 1.0 μM and 10 μM on viability.

TABLE-US-00001 TABLE 1 Viability Assay: Effect of Selected Compounds on Viability % viability Compound # Structure 10 μM 1.0 μM  9 (I-1) OBD9 [00027] 34.3 34.9 12 (I-2) [00028] 35.2 94.5 13 (I-3) [00029] 49.1 80.9 15 (I-4) [00030] 35.3 97.4 16 (I-5) [00031] 90.3 90.1 17 (I-6) OBD17 [00032] 84.8 93.9 51 (I-7) [00033] 54 (I-8) [00034] 58 (I-9) [00035] 66  (I-10) [00036]

Preparation of the Compounds

[0165] The compounds of Formula (I) provided herein can be prepared from readily available starting materials using the following general methods and procedures (e.g., Examples 1 to 9). Where typical or preferred process conditions (i.e., reaction temperatures, times, mole ratios of reactants, solvents, pressures, etc.) are given, other process conditions can also be used unless otherwise stated. Optimum reaction conditions may vary with the particular reactants or solvents used, but such conditions can be determined by those skilled in the art by routine optimization procedures.

Example 1. Preparation of Methyl N-(5-{4-[methyl(oxetan-3-yl)amino]benzoyl}-1H-1,3-benzodiazol-2-ylcarbamate

[0166] ##STR00037##

N-[4-(3,4-dinitrobenzoyl)phenyl]-N-methylacetamide

[0167] ##STR00038##

[0168] To a solution of 3,4-dinitrobenzoic acid (750 mg, 3.54 mmol) in DCM (8 mL) was added (COCl.sub.2).sub.2 (2.04 g, 16.08 mmol) and a drop of DMF at 0° C.; the resulting mixture was stirred at room temperature for 1 h, and thin layer chromatography (TIC) showed the reaction was over. The reaction mixture was concentrated to give an oil. Then, N-methyl-N-phenylacetamide (0.479 g, 3.21 mmol) and AlCl.sub.3 (1.6 g) was added, and the resulting mixture was stirred at 100° C. for 4 h. The reaction mixture was diluted with water, extracted with DCM, dried over Na.sub.2SO.sub.4, concentrated and purified by silica gel (PE:ethyl acetate=1:1) to give a yellow oil (240 mg, 22% yield). .sup.1H NMR (400 MHz, CDCl.sub.3): δ 8.33 (1H), 8.17 (1H), 8.05 (d, 1H), 7.85 (d, 2H), 7.42 (d, 2H), 3.35 (s, 3H), 2.05 (s, 3H). MS (ES): 344.2 (M+H).sup.+.

4-(3,4-dinitrobenzoyl)-N-methylaniline

[0169] ##STR00039##

[0170] To a solution of N-[4-(3,4-dinitrobenzoyl)phenyl]-N-methylacetamide (250 mg, 0.73 mmol) in water (2 mL) was added concentrated HCl (2 mmol) at 0° C., the resulting mixture was stirred at reflux overnight TLC showed the reaction was over. The reaction mixture was adjusted pH to 8 with aqueous NaOH, extracted with DCM, dried over Na.sub.2SO.sub.4, concentrated and purified by silica gel (PE:ethyl acetate=3:1) to afford the titled compound as a yellow solid (150 mg, 68% yield). .sup.1H NMR (400 MHz, CDCl.sub.3): δ 8.21 (1H), 8.05-8.0 (m, 21), 7.70 (d, 2H), 6.61 (d, 2H), 4.5 (br s, 1H), 2.95 (d, 3H). MS (ES): 302.3 (M+H).sup.+.

(3,4-dinitrophenyl)[4-(methylamino)phenyl]methanol

[0171] ##STR00040##

[0172] To a solution of 4-(3,4-dinitrobenzoyl)-N-methylaniline (500 mg, 0.166 mmol) in MeOH (8 mL) was added NaBH.sub.4 (123 mg, 3.22 mmol). The resulting mixture was stirred at room temperature for 1 h. TLC showed the reaction was over. The reaction was diluted with water, extracted with DCM, dried over Na.sub.2SO.sub.4, and concentrated to give the titled compound as a solid. (500 mg, 100% yield). .sup.1H NMR (400 MHz, CDCl.sub.3): δ 7.94 (1H), 7.88 (d, 1H), 7.72 (d, 1H), 7.11 (d, 2H), 6.58 (d, 2H), 5.80 (br s, 1H), 3.70 (br s, 1H), 2.83 (d, 3H). MS (ES): 304.0 (M+H).sup.+.

(3,4-dinitrophenyl)({4-[methyl(oxetan-3-yl)amino]phenyl})methanol

[0173] ##STR00041##

[0174] To a solution of (3,4-dinitrophenyl)[4-(methylamino)phenyl]methanol (50 mg, 0.166 mmol) in MeOH (1.5 mL) was added oxetan-3-one (30 mg, 0.415 mmol) and acetic acid (21 mg, 0.332 mmol). The resulting mixture was stirred at room temperature for 1 h. Then, NaBH.sub.3(CN) (36 mg, 0.332 mmol) was added to the mixture and stirred at room temperature overnight. LC-MS (Lot #: MC0968-024-R1) showed desired product was found. The reaction was diluted with water, extracted with DCM, dried over Na.sub.2SO.sub.4, concentrated and purified by silica gel to give the titled compound as a solid. (22 mg, 38% yield). .sup.1H NMR (400 MHz, CDCl.sub.3): δ 7.91 (m, 2H), 7.73 (d, 1H), 7.16 (d, 2H), 6.60 (d, 2H), 5.83 (d, 1H), 4.85 (m, 2H), 4.70 (m, 3H), 2.94 (s, 3H). MS (ES): 360.1 (M+H).sup.+.

N-[4-(3,4-dinitrobenzoyl)phenyl]-N-methyloxetan-3-amine

[0175] ##STR00042##

[0176] To a solution of (3,4-dinitrophenyl)({4-[methyl(oxetan-3-yl)amino]phenyl})methanol (250 mg, 0.696 mmol) in DCM (5 mL) was added DMP (590 mg, 1.39 mmol). The resulting mixture was stirred at room temperature for 1 h. The reaction was diluted with water, extracted with DCM, dried over Na.sub.2SO.sub.4, concentrated and purified by silica gel (PE:ethyl acetate=1:1) to give the titled compound as a yellow solid. (0.130 g, 52% yield). MS (ES): 358.1 (M−H).sup.−.

4-{4-[methyl(oxetan-3-yl)amino]benzoyl}benzene-1,2-diamine

[0177] ##STR00043##

[0178] To a solution of N-[4-(3,4-dinitrobenzoyl)phenyl]-N-methyloxetan-3-amine (100 mg, 0.280 mmol) in MeOH (1.5 mL) and THF (I mL) was added Pd/C (10 mg. wet, 10%). The mixture was stirred at room temperature for 1 h. The reaction mixture was filtered and the filtrate was concentrated to give the titled compound as a solid (Lot: mc0968-033-1, 90 mg, 100% yield, confirmed by HNMR). .sup.1H NMR (400 MHz, CDCl.sub.3): δ 7.74 (d, 2H), 7.25-7.22 (m, 2H), 6.70 (d, 1H), 6.60 (d, 2H). MS (ES): 298.1 (M+H).sup.+.

Methyl N-(5-(4-[methyl(oxetan-3-yl)amino]benzoyl)-1H-1,3-benzodiazol-2-yl)carbamate

[0179] ##STR00044##

[0180] To a solution of 4-(4-[methyl(oxetan-3-yl)amino]benzoyl}benzene-1,2-diamine (90 mg, 0.303 mmol) in EtOH (2 mL) was added 1,3-bis(methoxycarbonyl)-2-methyl-2-thiopseudourea (80 mg, 0.394 mmol). The mixture was stirred at reflux overnight. TLC showed the reaction was completed. The reaction mixture was concentrated and purified by silica gel (DCM:MeOH=15:1) to give the titled compound as a white solid (12 mg, 10% yield, confirmed by LC-MS, HNMR and HPLC). .sup.1H NMR (400 MHz, CDCl.sub.3) 3.09 (s, 3H), 3.88 (s, 3H), 4.81-4.92 (m, 5H), 6.64 (d, 2H), 7.44-8.11 (m, 6H), 10.74 (brs, 1H). MS (ES): 281.1 (M+H).sup.+.

Example 2. Preparation of (3-methyloxetan-3-yl)methyl N-[5-(propylsulfanyl)-1H-1,3-benzodiazol-2-yl]carbamate

[0181] ##STR00045##

[0182] The mixture of albendazole (530 mg) and 3-hydroxyl-3-methyl-oxetane (1 mL) in triethylamine (15 mL) was heated at 90° C. overnight. The solvent was removed under vacuum. The desired product was isolated by silica gel chromatography (hexanes:ethyl acetate=1:1) as a white solid. MS (ES): 336.5 (M+H).sup.+.

Example 3. Preparation of Oxetan-3-yl-N-(5-benzoyl-1H-1,3-benzodiazol-2-yl)carbamate

[0183] ##STR00046##

[0184] The mixture of mebendazole (295 mg), 3-hydroxyloxetane (370 mg), and triethylamine (0.5 mL) in DMF (15 mL) was heated in a microwave synthesizer at 60° C. for 15 minutes, then 70° C. for 5 minutes. The solvent was removed under vacuum. The desired product was isolated by silica gel chromatography (dichloromethane:methanol=3:1) as a white solid (180 mg, 53%). MS (ES): 338.5 (M+H).sup.+.

Example 4. Preparation of Oxetan-3-yl-N-[5-(phenylsulfanyl)-1H-1,3-benzodiazol-2-yl]carbamate

[0185] ##STR00047##

[0186] The mixture of fenbendazole (299 mg), 3-hydroxyloxetane (370 mg), and triethylamine (0.5 mL) in DMF (15 mL) was heated in a microwave synthesizer at then 70° C. for 5 minutes. The solvent was removed under vacuum. The desired product was isolated by silica gel chromatography (dichloromethane:methanol=9:1) as a white solid. MS (ES): 342.4 (M+H).sup.+.

Example 5. Preparation of Oxetan-3-yl N-[5-(propylsulfanyl)-JH-1,3-benzodiazol-2-yl]carbamate

[0187] ##STR00048##

[0188] The mixture of albendazole (530 mg) and 3-hydroxyloxetane (740 mg) in triethylamine was heated at 90° C. overnight. The solvent was removed under vacuum. The desired product was isolated by silica gel chromatography (dichloromethane:methanol=40:1) as a white solid. MS (ES): 308.6 (M+H).sup.+.

Example 5. Preparation of 2-(Dimethylamino)ethyl N-[5-(propylsulfanyl)-1H-1,3-benzodiazol-2-yl]carbamate

[0189] ##STR00049##

[0190] The mixture of albenzadole (265 mg, 1.0 mmol) an 2-(N,N-dimethylamino)ethanol (2.0 mL) in pyridine (15 mL) was heated to 90-100° C. for 3 hours. The desired product was isolated by silica gel chromatography (dichloromethane:methanol 3:1) as a yellow solid. MS (ES): 323.6 (M+H).sup.+.

Example 6. Preparation of 3-(dimethylamino)propyl N-[5-(propylsulfanyl)-1H-1,3-benzodiazol-2-yl]carbamate

[0191] ##STR00050##

[0192] The mixture of albenzadole (530 mg, 2.0 mmol) and 3-(N,N-dimethylamino)propanol (2.0 mL) in pyridine (15 mL) was heated to 90-100° C. for 3 hours. The desired product was isolated by silica gel chromatography (dichloromethane:methanol=3:1) as a yellow solid. MS (ES): 337.6 (M+H).sup.+.

Example 7. Preparation of 3-(dimethylamino)propyl N-[5-(4-fluorobenzoyl)-1H-1,3-benzodiazol-2-yl]carbamate

[0193] ##STR00051##

[0194] The mixture of flubendazole (626 mg, 2.0 mmol) and 3-(N,N-dimethylamino)propanol (2.0 mL) in pyridine (15 mL) was heated to 90-100° C. for 3 hours. The desired product was isolated by silica gel chromatography (dichloromethane:methanol=2:1) as a solid. MS (ES): 385.6 (M+H).sup.+.

Example 8. Preparation of methyl N-{5-[3-(4-methylpiperazin-1-yl)phenoxy]-1H-1,3-benzodiazol-2-yl}carbamate

[0195] ##STR00052##

3-(4-methylpiperazin-1-yl)phenol

[0196] ##STR00053##

[0197] To a stirred solution of 3-bromophenol (0.6 g, 3.5 mmol) in anhydrous toluene (12 mL) was added N-methylpiperazine (0.5 mL, 4.2 mmol), Pd.sub.2(dba), (79 mg, 0.087 mmol), .sup.tBuONa (0.933 g, 9.7 mmol) and BINAP (0.216 g, 0.35 mmol) in this sequence. The resulting solution was heated to reflux under the protection of argon for 1 hour. HPLC indicated the reaction is complete. After cooling to room temperature (rt), 20 mL of ethyl acetate and 20 mL of water were added to the reaction mixture, and the black precipitate was filtered. After separation of the filtrate, the water phase was extracted with ethyl acetate (20 mL×5). The combined organic phase was washed with brine, dried over Na.sub.2SO.sub.4, and concentrated under reduced pressure to give the crude product, which was purified by column chromatography (DCM:MeOH=20:1˜15:1˜10:1) to give the titled compound (376 mg, 56%).

5-[3-(4-methylpiperazin-1-yl)phenoxy]-2-nitroaniline

[0198] ##STR00054##

[0199] To a stirred solution of 3-(4-methylpiperazin-1-yl)phenol (360 mg, 1.87 mmol) in anhydrous DMF (1.5 mL) was added 60% NaH (54 mg, 2.24 mmol) slowly, followed by 5-chloro-2-nitroaniline (323 mg, 1.87 mmol) under argon. The resulting mixture was heated to 120° C. and kept overnight. The reaction was not yet complete indicated by TLC. Additional 5-chloro-2-nitroaniline (968 mg) and NaH (135 mg) were added to the reaction mixture. The reaction was complete after one hour. After cooling, 30 mL of cold water and 30 ml of ethyl acetate were added to the reaction mixture. The organic layer was separated and the aqueous layer was extracted with ethyl acetate twice (30 mL, 20 mL). The combined organic phase was washed with brine, dried over Na.sub.2SO.sub.4, and filtered. After solvent removal, purification on a silica gel column chromatography (DCM:MeOH=50:1˜40:1˜30:1˜20:1˜10:1) was used to afford the titled compound (406 mg, 66%).

4-[3-(4-methylpiperazin-1-yl)phenoxy]benzene-1,2-diamine

[0200] ##STR00055##

[0201] The substrate 5-[3-(4-methylpiperazin-1-yl)phenoxy]-2-nitroaniline (297 mg, 0.9 mmol) was treated with zinc dust (414 mg, 6.3 mmol) in acetic acid (19 ml) for one hour at room temperature (rt). The mixture was filtered through celite and the filter cake was washed with DCM. The filtrate was then concentrated under reduced pressure. 5N NaOH was added to the residue to pH 13, followed by DCM. The resulting precipitate was filtered and dried in vacuo to give the first batch product (72 mg). The filtrate was separated and the aqueous layer was extracted with DCM. The combined organic phase was dried over Na.sub.2SO.sub.4, concentrated under reduced pressure to give the second batch product (79 mg). HPLC indicated the two batches of product have similar purity. The two batches of product were combined (151 mg, 42%) for the next step.

Methyl N-{5-[3-(4-methylpiperazin-1-yl)phenoxy]-1H-1,3-benzodiazol-2-yl}carbamate

[0202] ##STR00056##

[0203] To a stirred solution of 4-[3-(4-methylpiperazin-1-yl)phenoxy]benzene-1,2-diamine (139 mg, 0.46 mmol) in acetic acid (4 mL) was added 1,3-bis(methoxycarbonyl)-2-methyl-2-thiopseudourea (240 mg, 1.16 mmol) under argon. The resulting solution was heated to 85° C. and kept for one hour. HPLC indicated the reaction is complete. After cooling to room temperature (rt), the solvent was removed under reduced pressure. 9 mL of 2N HCl was added to the residue and the mixture was stirred at room temperature for one hour. The mixture was then neutralized to pH 13 with aqueous 5N NaOH solution. The resulting aqueous phase was extracted with ethyl acetate three times. The combined organic phase was washed with brine, dried over Na.sub.2SO.sub.4. After solvent removal, sequence purification on a silica gel column chromatography (DCM:MeOH=50:1˜40:1˜30:1˜20:1˜10:1) was used to afford the titled compound as a pale yellow solid. .sup.1H NMR (400 MHz, CD.sub.6OD): δ 7.40 (m, 1H), 7.20 (m, 1H), 7.09 (s, 1H), 6.86 (d, 1H), 6.71 (d, 1H), 6.60 (s, 1H), 6.45 (d, 1H), 3.86 (s, 3H), 3.22 (m, 4H), 2.75 (m, 4H), 2.46 (s, 3H). MS (ES): 382.2 (M+H).sup.+.

Example 9. Preparation of methyl N-(5-{[6-(4-methylpiperazin-1-yl)pyridin-2-yl]oxy}-1H-1,3-benzodiazol-2-yl)carbamate

[0204] ##STR00057##

4-[(6-chloropyridin-2-yl)oxy]-2-nitroaniline

[0205] ##STR00058##

[0206] To a stirred solution of 2,6-dichloropyridine (0.48 g, 3.24 mmol), 4-amino-3-nitrophenol (0.5 g, 3.24 mmol) in anhydrous DMF (15 mL) was added potassium carbonate (0.67 g, 4.87 mmol) under argon. The resulting mixture was heated to 120° C. overnight. After cooling, 35 mL of water and 35 mL of ethyl acetate were added to the reaction mixture. After separation of the organic layer, the aqueous layer was extracted with ethyl acetate twice (100 mL, 50 mL). The combined organic phase was washed with brine, dried over Na.sub.2SO.sub.4, and concentrated under reduced pressure to give the titled compound (750 mg, 87%).

4-{[6-(4-methylpiperazin-1-yl)pyridin-2-yl]oxy}-2-nitroaniline

[0207] ##STR00059##

[0208] The substrate 4-[(6-chloropyridin-2-yl)oxy]-2-nitroaniline (0.44 g, 1.5 mmol) was dissolved in 8.8 mL of N-methylpiperazine. The resulting solution was heated to reflux under argon for three hours. After cooling to room temperature (rt), 30 mL of aqueous saturated NH.sub.4Cl solution was added slowly to the mixture. The resulting mixture was then extracted with ethyl acetate (20 mL×4). The combined organic phase was dried over Na.sub.2SO.sub.4, and concentrated under reduced pressure to give the titled compound for the next step.

4-{[6-(4-methylpiperazin-1-yl)pyridin-2-yl]oxy}benzene-1,2-diamine

[0209] ##STR00060##

[0210] The substrate 4-{[6-(4-methylpiperazin-1-yl)pyridin-2-yl]oxy}-2-nitroaniline (595 mg, 1.8 mmol) was treated with zinc dust (827 mg, 12.6 mmol) in acetic acid (39 mL) for 10 hours at room temperature (rt) The mixture was filtered through celite and the filter cake was washed with ethyl acetate. The filtrate was then concentrated under reduced pressure. 29 mL of 5N NaOH was added to the residue, followed by 20 ml of ethyl acetate. The resulting precipitate was filtered and dried in vacuo to give the first batch product (395 mg). The filtrate was separated and the aqueous layer was extracted with ethyl acetate (20 mL×2). The combined organic phase was dried over Na.sub.2SO.sub.4, and concentrated under reduced pressure to give the second batch product (267 mg). HPLC indicated the two batch product have similar purity. The two batches of product was combined (654 mg) for the next step.

Methyl N-(5-{[6-(4-methylpiperazin-1-yl)pyridin-2-yl]oxy}-1H-1,3-benzodiazol-2-yl)carbamate

[0211] ##STR00061##

[0212] To a stirred solution of 4-{[6-(4-methylpiperazin-1-yl)pyridin-2-yl]oxy}benzene-1,2-diamine (290 mg, 0.97 mmol) in acetic acid (8 mL) was added 1,3-bis(methoxycarbonyl)-2-methyl-2-thiopseudourea (499 mg, 2.42 mmol) under argon. The resulting solution was heated to 85° C. and kept for 30 min. HPLC indicated the reaction is complete. After cooling to room temperature (rt), the solvent was removed under reduced pressure. 15 mL of 2N HCl was added to the residue and the mixture was stirred at room temperature (rt) for one hour. The mixture was then neutralized to pH 13 with aqueous 5N NaOH solution. The resulting aqueous phase was extracted with ethyl acetate three times. The combined organic phase was dried over Na.sub.2SO.sub.4, and concentrated under reduced pressure to give the crude product, which was purified by sequential column chromatography (DCM:MeOH=50:1˜40:1˜30:1˜20:1˜10:1) to afford the titled compound (94 mg, 25%) as a pale yellow solid. .sup.1H NMR (400 MHz, CD.sub.6OD): δ 7.65 (m, 2H), 7.40 (s, 1H), 7.24 (d, 1H), 6.62 (d, 1H), 6.40 (d, 1H), 4.24 (d, 2H), 3.98 (s, 3H), 3.50 (m, 2H), 3.10 (m, 4H), 2.90 (s, 3H). MS (ES): 383.2 (M+H).sup.+.

EQUIVALENTS AND SCOPE

[0213] In the claims, articles such as “a,” “an,” and “the” may mean one or more than one unless indicated to the contrary or otherwise evident from the context. Claims or descriptions that include “or” between one or more members of a group are considered satisfied if one, more than one, or all of the group members are present in, employed in, or otherwise relevant to a given product or process unless indicated to the contrary or otherwise evident from the context. The invention includes embodiments in which exactly one member of the group is present in, employed in, or otherwise relevant to a given product or process. The invention includes embodiments in which more than one, or all of the group members are present in, employed in, or otherwise relevant to a given product or process.

[0214] Furthermore, the invention encompasses all variations, combinations, and permutations in which one or more limitations, elements, clauses, and descriptive terms from one or more of the listed claims is introduced into another claim. For example, any claim that is dependent on another claim can be modified to include one or more limitations found in any other claim that is dependent on the same base claim. Where elements are presented as lists, e.g., in Markush group format, each subgroup of the elements is also disclosed, and any element(s) can be removed from the group. It should it be understood that, in general, where the invention, or aspects of the invention, is/are referred to as comprising particular elements and/or features, certain embodiments of the invention or aspects of the invention consist, or consist essentially of, such elements and/or features. For purposes of simplicity, those embodiments have not been specifically set forth in haec verba herein. It is also noted that the terms “comprising” and “containing” are intended to be open and permits the inclusion of additional elements or steps. Where ranges are given, endpoints are included. Furthermore, unless otherwise indicated or otherwise evident from the context and understanding of one of ordinary skill in the art, values that are expressed as ranges can assume any specific value or sub-range within the stated ranges in different embodiments of the invention, to the tenth of the unit of the lower limit of the range, unless the context clearly dictates otherwise.

[0215] This application refers to various issued patents, published patent applications, journal articles, and other publications, all of which are incorporated herein by reference. If there is a conflict between any of the incorporated references and the instant specification, the specification shall control. In addition, any particular embodiment of the present invention that falls within the prior an may be explicitly excluded from any one or more of the claims. Because such embodiments are deemed to be known to one of ordinary skill in the art, they may be excluded even if the exclusion is not set forth explicitly herein. Any particular embodiment of the invention can be excluded from any claim, for any reason, whether or not related to the existence of prior art.

[0216] Those skilled in the art will recognize or be able to ascertain using no more than routine experimentation many equivalents to the specific embodiments described herein. The scope of the present embodiments described herein is not intended to be limited to the above Description, but rather is as set forth in the appended claims. Those of ordinary skill in the art will appreciate that various changes and modifications to this description may be made without departing from the spirit or scope of the present invention, as defined in the following claims.