HK1238131A1 - Compositions for treating cancers - Google Patents

Abstract

The invention describes compounds that inhibit both HDAC and GSK3β (i.e., HDAC/GSK3β dual inhibitors). The invention further describes compositions containing these HDAC/GSK3β dual inhibitors, as well as methods and kits using these HDAC/GSK3β dual inhibitors to treat various medical conditions. The invention also provides methods and kits using a HDAC inhibitor and a GSK3β to treat various medical conditions, and compositions containing a HDAC inhibitor and a GSK3β. Medical conditions treatable with various embodiments of the invention include but are not limited to caners and tumors.

Description

Compositions and methods for treating cancer

Cross Reference to Related Applications

This application claims the benefit of united states provisional application No. 62/011,413, filed 2014, 12/6/119 (e) requirements 2014, which is hereby incorporated by reference in its entirety.

Statement regarding federally sponsored research

The invention was made with government support under grant numbers AA019996 and CA163200 awarded by the National Institutes of Health (NIH). The government has certain rights in this invention.

Technical Field

The present invention relates to compounds, compositions, methods and kits for treating medical conditions. Such disorders include, but are not limited to, cancer or tumors.

Background

All publications cited herein are incorporated by reference in their entirety to the same extent as if each individual publication or patent application was specifically and individually indicated to be incorporated by reference. The following description includes information that may be useful in understanding the present invention. It is not an admission that any of the information provided herein is prior art or relevant to the presently claimed invention, or that any publication specifically or implicitly referenced is prior art.

In humans, there is a strong correlation between overexpression of glycogen synthase kinase 3 β (GSK3 β) and cancer progression. Activation of GSK3 β upregulates cancer cell proliferation and increases cancer cell resistance to apoptosis by activating pro-survival pathways, including the NF- κ B pathway. These observations suggest that inhibition of GSK3 β is a potential therapeutic strategy for many cancers. However, although GSK3 β inhibitors reduce cancer cell proliferation, they stimulate the transformation of cancer cells into cells that are more likely to invade surrounding normal tissues and metastasize. This transformation to an invasive transition state is called epithelial-mesenchymal transition (EMT). EMT is also associated with cancer cells that are more resistant to treatment due to their transformation into cancer stem cells (or sternness).

The present invention demonstrates that inhibitors of the enzyme Histone Deacetylase (HDAC) prevent EMT and enhance the anti-tumor effect of GSK3 β inhibitors, and provides compounds, compositions, methods and kits for treating a variety of conditions, including but not limited to cancer and tumors.

Summary of The Invention

Various embodiments of the present invention provide compounds that inhibit both HDAC and GSK3 β (i.e., dual inhibitors of HDAC and GSK3 β). In some embodiments, the dual inhibitor compound is represented by (V):

wherein: l is₁And L₂Independently a linker; r¹Is an aromatic moiety, alkyl, acyl, cyclyl or heterocyclyl, each of which may be optionally substituted; r²Is hydrogen, lower alkyl, cyclyl, heterocyclyl, aryl or heteroaryl, each of which may be optionally substituted; r³Is absent or is an aromatic moiety, which may be optionally substituted; p is 0, 1, 2, 3, 4, 5, 6,7, 8, 9 or 10; and one of them. R¹-L₁-one nitrogen (nitorgen) attached to the thiadiazolidine ring, and- (CH)₂)_p-R³-L₂-C(O)NHOR²Another nitrogen (nitorgen) attached to the thiadiazolidine ring.

In some other embodiments, the dual inhibitor compound is represented by formula (V):

wherein: l is₁And L₂Independently a linker; r¹Is an aromatic moiety, alkyl, acyl, cyclyl or heterocyclyl, each of which may be optionally substituted; r²Is hydrogen, lower alkyl, cyclyl, heterocyclyl, aryl or heteroaryl, each of which may be optionally substituted; r³Is absent or is an aromatic moiety, which may be optionally substituted; and p is 0, 1, 2, 3, 4, 5, 6,7, 8, 9 or 10.

Various embodiments of the present invention provide compositions consisting of, consisting essentially of, or comprising dual inhibitors of HDAC and GSK3 β. In various further embodiments, the dual inhibitor is attached to a cleavable enzyme substrate. In some embodiments, the cleavable enzyme substrate is attached to a particle, such as a magnetic particle.

Various embodiments of the present invention provide methods of treating a disorder, preventing a disorder, reducing the likelihood of developing a disorder, reducing the severity of a disorder, and/or slowing the progression of a disorder in a subject. The method consists of, or consists essentially of, or comprises: administering to the subject a therapeutically effective amount of a dual inhibitor of HDAC and GSK3 β, thereby treating, preventing, reducing the likelihood of, reducing the severity of, and/or slowing the progression of the disorder in the subject. In various embodiments, the method further comprises providing a dual inhibitor.

In various embodiments, in addition to administering the dual inhibitor, the method may further comprise administering or treating with one or more additional anti-cancer therapies. In some such embodiments, the additional anti-cancer therapy comprises surgery, radiation therapy, biological therapy, immunotherapy, chemotherapy, or any combination thereof.

In addition to administering the dual inhibitor, some embodiments of the methods may further comprise administering or treating with one or more anti-cancer therapeutic agents. In some such embodiments, the anti-cancer therapeutic agent can be a chemotherapeutic agent, a growth inhibitory agent, an anti-angiogenic agent, a cytotoxic agent, an anti-hormonal agent, a prodrug, a cytokine, or any combination thereof. In some embodiments, the method further comprises administering a chemotherapeutic agent to the subject.

In further embodiments, the dual inhibitor is attached to a cleavable enzyme substrate, and the cleavable enzyme substrate is attached to a magnetic particle, and the method further comprises directing the dual inhibitor to the cancer or tumor using a magnetic field.

Various embodiments of the present invention provide kits for treating a disorder, preventing a disorder, reducing the severity of a disorder, and/or slowing the progression of a disorder in a subject. The kit consists of or consists essentially of or comprises: an amount of a dual inhibitor of HDAC and GSK3 β; and instructions for using the dual inhibitor to treat, prevent, reduce the likelihood of, reduce the severity of, and/or slow the progression of a disorder in a subject. In various further embodiments, the dual inhibitor is attached to a cleavable enzyme substrate, and the cleavable enzyme substrate is attached to the magnetic particle.

Various embodiments of the present invention provide compositions consisting of, consisting essentially of, or comprising an HDAC inhibitor and a GSK3 β inhibitor. In various further embodiments, the HDAC inhibitor and/or GSK3 β inhibitor is attached to a cleavable enzyme substrate, and the cleavable enzyme substrate is attached to a magnetic particle.

Various embodiments of the present invention provide methods of treating a disorder, preventing a disorder, reducing the likelihood of developing a disorder, reducing the severity of a disorder, and/or slowing the progression of a disorder in a subject. The method consists of, or consists essentially of, or comprises: administering to the subject a therapeutically effective amount of an HDAC inhibitor and a GSK3 β inhibitor, thereby treating, preventing, reducing the likelihood of, reducing the severity of, and/or slowing the progression of the disorder in the subject. In various embodiments, the method further comprises providing an HDAC inhibitor and/or a GSK3 β inhibitor.

In various embodiments, the methods may further comprise administering or treating with one or more additional anti-cancer therapies in addition to the HDAC inhibitor and GSK3 β inhibitor. In some such embodiments, the additional anti-cancer therapy comprises surgery, radiation therapy, biological therapy, immunotherapy, chemotherapy, or any combination thereof.

In addition to the HDAC inhibitor and GSK3 β inhibitor, some embodiments of the methods may further comprise administering or treating with one or more anti-cancer therapeutic agents. In some such embodiments, the anti-cancer therapeutic agent can be a chemotherapeutic agent, a growth inhibitory agent, an anti-angiogenic agent, a cytotoxic agent, an anti-hormonal agent, a prodrug, a cytokine, or any combination thereof. In some embodiments, the method further comprises administering a chemotherapeutic agent to the subject.

In other embodiments, the HDAC inhibitor and/or GSK3 β inhibitor is attached to a cleavable enzyme substrate, and the cleavable enzyme substrate is attached to a magnetic particle, and the method further comprises using a magnetic field to direct the HDAC inhibitor and/or GSK3 β inhibitor to the cancer or tumor.

Various embodiments of the present invention provide kits for treating a disorder, preventing a disorder, reducing the severity of a disorder, and/or slowing the progression of a disorder in a subject. The kit consists of or consists essentially of or comprises: an amount of an HDAC inhibitor; an amount of a GSK3 β inhibitor; and instructions for using the HDAC inhibitor and the GSK3 β inhibitor to treat, prevent, reduce the likelihood of, reduce the severity of, and/or slow the progression of a disorder in a subject. In various further embodiments, the HDAC inhibitor and/or GSK3 β inhibitor is attached to a cleavable enzyme substrate, and the cleavable enzyme substrate is attached to a magnetic particle.

Various compounds, compositions, methods and kits of the invention are useful for treating a variety of conditions, including but not limited to cancer and tumors.

Drawings

Exemplary embodiments are shown in the referenced figures. The embodiments and figures disclosed herein are intended to be considered illustrative rather than restrictive.

Figure 1 depicts the inventors' novel strategy for inhibiting cancer growth, metastasis and resistance to treatment, according to various embodiments of the present invention.

Figure 2 depicts the effect of the HDAC inhibitor Saha (50mg/Kg) and GSK3 β inhibitor (4mg/Kg), alone and in combination, on reducing PanIN lesion formation, according to various embodiments of the present invention.

Figure 3 depicts the effect of the HDAC inhibitor Saha (50mg/Kg) and GSK3 β inhibitor (4mg/Kg), alone and in combination, on fibrosis, according to various embodiments of the present invention.

Fig. 4 depicts the effect of HDAC, GSK3 β inhibition in combination with gemcitabine on EMT in MIA PaCa-2 pancreatic cancer cells, according to various embodiments of the present invention.

Fig. 5 depicts the dose-dependent effect of the GSK3 β inhibitor tideglusib on cell survival in pancreatic cancer cells MIA PaCa-2 (p < 0.05 compared to control), according to various embodiments of the invention.

Figure 6 depicts the dose-dependent effect of the HDAC inhibitor Saha on cell survival in pancreatic cancer cells MIA PaCa-2 (p < 0.05 compared to control), according to various embodiments of the present invention.

Fig. 7 depicts the effect of a combination of the GSK3 β inhibitor tideglusib and the HDAC inhibitor Saha on cell survival in pancreatic cancer cells MIA PaCa-2, according to various embodiments of the present invention. (and p < 0.05 compared to Saha alone).

Fig. 8 depicts the structure of an example of an HDAC inhibitor according to various embodiments of the present invention. For example, suberoylanilide hydroxamic acid (SAHA) binds to the active site of HDAC and acts as a chelator for zinc ions also present in the active site of HDAC. More information can be found in Ekou et al (stone Deacetylase Inhibitors: Synthesis of polypeptide Analogue SAHA/TPX; J.chem.2011; 8 (S1): S79-S84), the entire contents of which are incorporated herein by reference as if fully set forth.

Fig. 9 depicts the structure of an example of a GSK3 β inhibitor, according to various embodiments of the invention. For example, SB216763 is an ATP analog; TDZD-8, a thiadiazolidinone derivative, is a potent selective small molecule non-ATP competitive GSK3 β inhibitor; and Tideglusib (NP-12; 4-benzyl-2- (naphthalen-1-yl) -1, 2, 4-thiadiazolidine-3, 5-dione) is a potent selective small molecule non-ATP competitive GSK3 β inhibitor.

Figure 10 depicts one non-limiting example of the inventors' compounds that inhibit both HDAC and GSK3 β, according to various embodiments of the present invention. Ar refers to an aromatic moiety and the spacer refers to a carbon linker.

Fig. 11 depicts one non-limiting example of a protease moiety for pancreatic cancer, according to various embodiments of the invention: cathepsin G substrate.

Fig. 12 depicts non-limiting examples of nanomaterials for pancreatic cancer, according to various embodiments of the invention: a siMAG. As shown, HDAC inhibitors, GSK3 β inhibitors and/or dual inhibitors may be conjugated to si-MAG particles using well-known carbodiimide coupling methods or mannich reactions (mannich).

FIG. 13 shows that compound ALB-185357 dose-dependently decreased cell survival as measured by the MTT assay in BxPC-3 pancreatic cancer cell line cultured for 72 hours. Significance compared to control; significance between tideglusib + saha and ALB-185357 used at the same concentration, p < 0.05.

FIG. 14 shows that ALB-185357 dose-dependently increased apoptosis as measured by the level of DNA fragmentation in the MIA PaCa-2 pancreatic cancer cell line. Significance compared to control, p < 0.05.

FIG. 15 shows that the combination of ALB-185357 and gemcitabine induced a synergistic effect on induction of apoptosis in the MIA PaCa-2 pancreatic cancer cell line. The dashed line indicates the expected additive effect (significance between the expected and observed effect achieved at 2.4 uM).

FIGS. 16A and 16B show that compounds ALB-188540 (FIG. 16A) and ALB-185643 (FIG. 16B) show similar effects on the survival of BxPC3 cells as compound ALB-185357. Significance compared to control, p < 0.05.

Figures 17-19 show that compound ALB-185357 dose-dependently reduced cell survival as measured by MTT assay and cell number in different cancers, including BT474 breast cancer cells (figure 17), hepatocellular carcinoma HepG2 cells (figure 18), and Raji lymphoma cells (figure 19). Significance compared to control, p < 0.05. This data indicates that ALB-185357 inhibits cell survival of multiple cancer cell types.

Figure 20 shows that compound ALB-185357 did not affect cell survival of normal pancreatic ductal cells. Significance compared to control, p < 0.05.

FIG. 21 shows that compound ALB-185357 dose-dependently upregulated predicted targets in the MIA PaCa-2 pancreatic cancer cell line: histone acetylation and GSK-3 β phosphorylation/inhibition.

Figures 22A and 22B show that compound ALB-185357 decreased expression of markers mediating epithelial-to-mesenchymal transition of metastases (N-cadherin and twist) and a cancer sternness marker mediating resistance to treatment (Sox2) (figure 22A) and decreased invasion of MIAPaCa-2 pancreatic cancer cell line (figure 22B).

Figure 23 shows that compound ALB-185357 significantly increased mouse survival by at least 50%.

Detailed Description

All references cited herein are incorporated by reference in their entirety as if fully set forth. Unless defined otherwise, technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Allen et al, Remington: the Science and Practice of Pharmacy 22 nd edition, pharmaceutical Press (9, 15, 2012); hornyak et al, Introduction to Nanoscience and nanotechnology, CRC Press (2008); singleton and Sainsbury, Dictionary of microbiology and Molecular Biology 3 rd edition, revision, J.Wiley&Sons (New York, NY 2006); smith, March's Advanced Organic Chemistry Reactions, mechanics and Structure 7 th edition, J.Wiley&Sons (New York, NY 2013); singleton, Dictionary of DNA and genome Technology 3 rd edition, Wiley-Blackwell (November 28, 2012); and Green and Sambrook, Molecular Cloning: a Laboratory Manual 4 th edition, Cold Spring Harbor Laboratory Press (Cold Spring Harbor, NY 2012) is available to those skilled in the artGeneral guidance for a number of terms used in this application is provided. For references on how to prepare Antibodies, see Greenfield, Antibodies laboratory Manual 2 nd edition, Cold Spring Harbor Press (Cold Spring Harbor NY, 2013);and Milstein, Derivation of specific antisense-reducing tissue culture and tumor lines by cell fusion, Eur.J.Immunol.1976, 7 months, 6 (7): 511-9; queen and Selick, Humanized immunoglobulins, U.S. Pat. No.5,585,089(1996, 12 months); and Riechmann et al, rehaping human antibodies for therapy, Nature 24/3 1988, 332 (6162): 323-7.

Those skilled in the art will recognize many methods and materials similar or equivalent to those described herein, which can be used in the practice of the present invention. Other features and advantages of the present invention will become apparent from the following detailed description, taken in conjunction with the accompanying drawings, which illustrate, by way of example, various features of embodiments of the invention. Indeed, the invention is in no way limited to the methods and materials described. For convenience, certain terms used in the specification, examples, and appended claims herein are collected.

Unless otherwise indicated or implied from the context, the following terms and phrases include the meanings provided below. The following terms and phrases do not exclude the meaning of the term or phrase as it is acquired in the art unless expressly stated otherwise or apparent from the context. These definitions are provided to help describe particular embodiments and are not intended to limit the claimed invention, as the scope of the invention is limited only by the claims. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

As used herein, the terms "comprising" or "comprises" are used to refer to compositions, methods, and respective components thereof that are useful for the embodiments, but are also open to inclusion of unspecified elements, whether useful or not. It will be understood by those within the art that, in general, terms used herein are generally intended as "open" terms (e.g., the term "including" should be interpreted as "including but not limited to," the term "having" should be interpreted as "having at least," the term "includes" should be interpreted as "includes but is not limited to," etc.).

The terms "a" and "an" and "the" used in the context of describing particular embodiments of the present application (especially in the context of the claims) may be construed to cover both the singular and the plural, unless otherwise indicated. Recitation of ranges of values herein are merely intended to serve as a shorthand method of referring individually to each separate value falling within the range. Unless otherwise indicated herein, each individual value is incorporated into the specification as if it were individually recited herein. All methods described herein can be performed in any suitable order unless otherwise indicated herein or otherwise clearly contradicted by context. The use of any and all examples, or exemplary language (e.g., "such as") provided with respect to certain embodiments herein, is intended merely to better illuminate the application and does not pose a limitation on the scope of the application claimed. The abbreviation "e.g." is derived from latin languages such as (exempli gratia) and is used herein to indicate non-limiting examples. The abbreviation "e.g." is therefore synonymous with the term "e.g". No language in the specification should be construed as indicating any non-claimed element as essential to the practice of the application.

As used herein, the terms "treat," "treatment," "treating," or "ameliorating," when referring to a disease, condition, or medical disorder, refer to both therapeutic treatment and prophylactic or preventative measures, wherein the object is to prevent, reverse, alleviate, ameliorate, inhibit, lessen, slow or stop the progression or severity of the symptoms or disorders. The term "treating" includes reducing or alleviating at least one adverse effect or symptom of a disorder. A treatment is typically "effective" if one or more symptoms or clinical markers are reduced. Alternatively, a treatment is "effective" if the progression of the disease, condition, or medical disorder is reduced or halted. That is, "treatment" includes not only improvement of symptoms or markers, but also stopping or at least slowing of progression or worsening of symptoms expected in the absence of treatment. Further, "treating" may mean pursuing or obtaining a beneficial result, or reducing the individual's chance of developing a condition, even if the treatment is ultimately unsuccessful. Those in need of treatment include those already with the disorder as well as those predisposed to the disorder or in which the disorder is to be prevented.

A "beneficial result" or "desired result" can include, but is in no way limited to, lessening or lessening the severity of a disease condition, preventing the worsening of a disease condition, curing a disease condition, preventing the progression of a disease condition, reducing the patient's chances of developing a disease condition, reducing morbidity and mortality, extending the life or life expectancy of the patient. By way of non-limiting example, a "beneficial result" or "desired result" can be alleviation of one or more symptoms, diminishment of extent of deficiency, stabilization (i.e., not worsening) of cancer or tumor status, delay or alleviation of cancer or tumor, and improvement or alleviation of symptoms associated with cancer or tumor.

As used herein, "disorder" and "disease disorder" may include, but are in no way limited to, any form of malignant neoplastic cell proliferative condition or disease. Examples of such conditions include, but are not limited to, cancer and tumors.

As used herein, "cancer" or "tumor" refers to the uncontrolled growth of cells that interfere with the normal function of body organs and systems, and/or the growth and proliferation of all neoplastic cells, whether malignant or benign, as well as all pre-cancerous and cancerous cells and tissues. A subject having a cancer or tumor is a subject having objectively measurable cancer cells present in the subject's body. Included in this definition are benign and malignant cancers, as well as dormant tumors or micrometastases. Cancers that migrate from their original location and implant vital organs may eventually lead to death of the subject by functional deterioration of the affected organs. As used herein, the term "invasiveness" refers to the ability to invade and destroy surrounding tissue. Melanoma is an aggressive form of skin tumor. As used herein, the term "cancer (carcinoma)" refers to a cancer produced by epithelial cells.

Examples of cancer include, but are not limited to, tumors of the nervous system, brain, nerve sheath, breast, colon, carcinoma, lung, hepatocellular, gastric, pancreatic, cervical, ovarian, liver, bladder, urinary tract, thyroid, kidney, renal cell, melanoma, head and neck, brain, and prostate (including, but not limited to, androgen-dependent and androgen-independent prostate cancer). Examples of brain tumors include, but are not limited to, benign brain tumors, malignant brain tumors, primary brain tumors, secondary brain tumors, metastatic brain tumors, gliomas, glioblastoma multiforme (GBM), medulloblastomas, ependymomas, astrocytomas, hairy cell astrocytomas, oligodendrogliomas, brain stem gliomas, optic gliomas, mixed gliomas such as oligodendroastrocytomas, low-grade gliomas (low-grade gliomas), high-grade gliomas (high-grade gliomas), supratentorias, subthreshold gliomas, pontine gliomas, meningiomas, pituitary adenomas, and schwannoma. Nervous system tumor or nervous system neoplasm refers to any tumor that affects the nervous system. The nervous system tumor may be a tumor in the Central Nervous System (CNS), the Peripheral Nervous System (PNS), or both the CNS and PNS. Examples of nervous system tumors include, but are not limited to, brain tumors, schwannoma, and optic glioma.

As used herein, the term "administering" refers to placing an agent disclosed herein into a subject by a method or route that results in at least partial localization of the agent at a desired site. The "route of administration" may refer to any route of administration known in the art, including, but not limited to, aerosol, nasal, oral, transmucosal, transdermal, parenteral, enteral, topical, or topical. "parenteral" refers to a route of administration typically associated with injection, including intraorbital, infusion, intra-arterial, intracapsular, intracardiac, intradermal, intramuscular, intraperitoneal, intrapulmonary, intraspinal, intrasternal, intrathecal, intrauterine, intravenous, subarachnoid, subcapsular, subcutaneous, transmucosal, or transtracheal. By parenteral route, the compositions may be in the form of solutions or suspensions for infusion or injection, or as lyophilized powders. By the enteral route, the pharmaceutical composition may be in the form of tablets, gel capsules, sugar-coated tablets, syrups, suspensions, solutions, powders, granules, emulsions, microspheres or nanospheres or lipid or polymer vesicles that allow controlled release. By topical route, the pharmaceutical composition may be in the form of an aerosol, lotion, cream, gel, ointment, suspension, solution or emulsion. According to the invention, "administering" may be self-administering. For example, a subject is considered to "administer" a composition as disclosed herein.

The term "sample" or "biological sample" as used herein means a sample obtained or isolated from a biological organism, for example a tumor sample from a subject. Exemplary biological samples include, but are not limited to, biological fluid samples; serum; plasma; (ii) urine; saliva; a tumor sample; tumor biopsy and/or tissue samples, etc. The term also includes mixtures of the above samples. The term "sample" also includes untreated or pretreated (or pre-processed) biological samples. In some embodiments, the sample may comprise one or more cells from the subject. In some embodiments, the sample may be a tumor cell sample, e.g., the sample may comprise cancer cells, cells from a tumor, and/or a tumor biopsy.

As used herein, "subject" refers to a human or an animal. Typically, the animal is a vertebrate, such as a primate, rodent, domestic animal or hunting animal. Primates include chimpanzees, cynomolgus monkeys, spider monkeys, and macaques, such as rhesus monkeys. Rodents include mice, rats, woodchucks, ferrets, rabbits, and hamsters. Domestic and hunting animals include cattle, horses, pigs, deer, bison, buffalo, felines such as domestic cats, and canines such as dogs, foxes, wolves. The terms "patient," "individual," and "subject" are used interchangeably herein. In one embodiment, the subject is a mammal. The mammal may be a human, non-human primate, mouse, rat, dog, cat, horse, or cow, but is not limited to these examples. In addition, the methods described herein may be used to treat domesticated animals and/or pets.

As used herein, "mammal" refers to any member of the mammalian class, including but not limited to humans and non-human primates, such as chimpanzees and other apes and monkey species; farm animals such as cattle, sheep, pigs, goats, and horses; domestic mammals such as dogs and cats; the experimental animals include rodents such as mice, rats, guinea pigs, and the like. The term does not denote a particular age or gender. Thus, adult and newborn subjects, as well as fetuses, whether male or female, are intended to be included within the scope of this term.

The subject may be a subject who has been previously diagnosed or identified as having or having a condition in need of treatment (e.g., a cancer or tumor) or one or more complications associated with the condition, and optionally has undergone treatment for the condition or one or more complications associated with the condition. Alternatively, the subject may also be a subject that has not been previously diagnosed as having a disorder or one or more complications associated with the disorder. For example, the subject may be a subject exhibiting a risk factor for the disorder or one or more complications associated with the disorder, or a subject not exhibiting a risk factor. A subject "in need of treatment" for a particular disorder can be a subject suspected of having, diagnosed as having, treated or treating, not treating, or at risk of developing the disorder.

The term "anti-cancer therapy" refers to a therapy useful for treating cancer. Examples of anti-cancer therapeutic agents include, but are not limited to, for example, surgery, radiation therapy, chemotherapeutic agents, growth inhibitory agents, cytotoxic agents, agents for radiation therapy, anti-angiogenic agents, apoptotic agents, anti-tubulin agents, and other agents for treating cancer, such as anti-HER-2 antibodies (e.g., Herceptin)^TM)、anti-CD 20 antibodies, Epidermal Growth Factor Receptor (EGFR) antagonists (e.g., tyrosine kinase inhibitors), HER1/EGFR inhibitors (e.g., erlotinib (Tarceva)^TM) Platelet derived growth factor inhibitors (e.g., Gleevec)^TM(imatinib mesylate)), COX-2 inhibitors (e.g., celecoxib), interferons, cytokines, antagonists (e.g., neutralizing antibodies) that bind to one or more of ErbB2, ErbB3, ErbB4, PDGFR- β, BlyS, APRIL, BCMA or VEGF receptors, TRAIL/Apo2 and other biologically active and organic chemical agents, and the like.

The term "cytotoxic agent" as used herein refers to a substance that inhibits or prevents cellular function and/or causes cellular destruction. The term is intended to include radioisotopes (e.g., At)²¹¹、I¹³¹、I¹²⁵、Y⁹⁰、Re¹⁸⁶、Re¹⁸⁸、Sm¹⁵³、Bi²¹²、p³²And radioactive isotopes of Lu), chemotherapeutic agents, and toxins of bacterial, fungal, plant, or animal origin, such as small molecule toxins or enzymatically active toxins, including fragments and/or variants thereof.

As used herein, a "chemotherapeutic agent" is a chemical compound useful in the treatment of cancer. Examples of chemotherapeutic agents include, but are not limited to: alkylating agents, e.g. thiotepa and cycloxan^TMCyclophosphamide; alkyl sulfonates such as busulfan, improsulfan and piposulfan; aziridines such as benzotepa (benzodopa), carboquone (carboquone), metotepipa (meturedpa), and uredepa (uredpa); ethyleneimine (ethylenimine) and methylmelamine (melamelamines), including altretamine (altretamine), tritylamine (triethyleneamine), triethylenephosphoramide (triethylenephosphoramide), triethylenethiophosphoramide (triethylenethiophosphamide), and trimethylolmelamine (trimethylomelamine); acetogenin (especially annonaceous acetogenin (bullatacin) and bullatacin (bullatacinone)); camptothecin (including the synthetic analog topotecan); bryostatins; a caristatin (callystatin); CC-1065 (including Alexanin: (A)adozelesin), kazelesin (carzelesin), and bizelesin (bizelesin) synthetic analogs); cryptophycin (cryptophycin) (especially cryptophycin 1 and cryptophycin 8); dolastatin (dolastatin); duocarmycin (duocarmycin) (including the synthetic analogs KW-2189 and CB1-TM 1); shogaol (eleutherobin); coprinus atrata base (pancratistatin); sarcandra glabra alcohol (sarcodictyin); spongistatin (spongistatin); nitrogen mustards (nitrogen mustards), such as chlorambucil (chlorambucil), chlorambucil (chlorenaphazine), chromyl phosphate (chlorophosphamide), estramustine (estramustine), ifosfamide (ifosfamide), mechlorethamine (mechlorethamine), mechlorethamine oxide hydrochloride, melphalan (melphalan), neonebivory (novembichin), benzene mustard (pheresteresterone), prednimustine (prednimustine), triamcinolone (trofosfamide), uracil mustard (uracil mustard); nitrosoureas (nitrosureas), such as carmustine (carmustine), chlorouretocin (chlorozotocin), fotemustine (fotemustine), lomustine (lomustine), nimustine (nimustine) and ramustine (ranirnustine); antibiotics, such as enediyne antibiotics (e.g., calicheamicin, particularly calicheamicin γ 1I and calicheamicin ω I1 (see, e.g., Agnew, chem. int. Engl. eds., 33: 183-186(1994)), dalnamycin (dynemicin), including dalnamycin A; bisphosphonates (bisphosphates), such as clodronate (clodronate); esperamicin (esperamicin), and neocarzinostain chromophores and related chromoprotein enediyne chromophores), aclacinomycin (acarinomysin), actinomycin (actinomycin), anidamycin (aurramycin), azamethicin (azamethicin), bleomycin (bleomycin), actinomycin C (cacinomycin), carubicin (carvacomycin), daunomycin (carminomycin), carminomycin (carubicin), daunomycin (gentamycin), daunomycin (gentin), daunomycin (gentin (gentamycin), daunomycin (gentin (gentamycin), daunomycin (gentin), daunomycin (gent), daunomycin (daunomycin), daunomycin (daunomycin), daunom, ADRIAMYCIN^TMDoxorubicin (doxorubicin) (including morpholino doxorubicin, cyanomorpholino doxorubicin, 2-pyrrolidinyl doxorubicin, and doxorubicin), epirubicin (epirubicin), isorubicin (esorubicin), idarubicin (idarubicin), and sisomicin (marce)llomycin), mitomycins (mitomycins) such as mitomycin C, mycophenolic acid (mycophenolic acid), nogomycin (nogalamycin), olivomycin (olivomycin), pellomycin (polyplomycin), pofiomycin (potfiromycin), puromycin (puromycin), triiron (quelamycin), rodobicin (rodorubicin), streptonigrin (streptonigrin), streptozotocin (streptozotocin), tubercidin (tubicidin), ubenimex (ubenimex), neocarzinostatin (zinostatin), zorubicin (zorubicin); antimetabolites such as methotrexate and 5-fluorouracil (5-FU); folic acid analogs such as denopterin, methotrexate, pteropterin, trimetrexate; purine analogs, such as fludarabine (fludarabine), 6-mercaptopurine, thiamidine (thiamiriprine), thioguanine (thioguanine); pyrimidine analogs, such as cyclocytidine (ancitabine), azacitidine (azacitidine), 6-azauridine, carmofur (carmofur), cytarabine (cytarabine), dideoxyuridine (dideoxyuridine), deoxyfluorouridine (doxifluridine), enocitabine (enocitabine), floxuridine; androgens such as testosterone carprofonate (calusterone), drostanolone propionate (dromostanolone propionate), epithioandrostanol (epithiostanol), mepiquitane (mepiquitazone), testolactone (testolactone); anti-adrenal agents, such as aminoglutethimide (aminoglutethimide), mitotane (mitotane), trilostane (trilostane); folic acid supplements, such as folinic acid (frilic acid); acetoglucurolactone (acegultone); (ii) an aldophosphamide glycoside; aminolevulinic acid (aminolevulinic acid); eniluracil (eniluracil); amsacrine (amsacrine); betribucin (betrabucil); a bartrene (bisantrene); edatrexate (edatraxate); desphosphamide (defofamine); colchicine (demecolcine); diazaquinone (diaziqutone); eflornithine (elformithine); ammonium etitanium acetate; epothilone (epothilone); etoglut (etoglucid); gallium nitrate; a hydroxyurea; lentinan; lonidanine (lonidanine); maytansinoids (maytansinoids), such as maytansine and maytansinoids (ansamitocins); mitoguazone (mitoguzone); mitoxantrone (mitoxantrone); mopidanol (mopidanmol); nitrerine (nitrerine); pentostatin (pentostatin); egg amineNitrogen mustard (phenamett); pirarubicin (pirarubicin); losoxantrone (losoxantrone); podophyllinic acid (podophyllic acid); 2-ethyl hydrazide (2-ethyl hydrazide); procarbazine (procarbazine); PSK^TMPolysaccharide complex (JHS Natural Products, Eugene, Oreg.); razoxane (rizoxane); rhizomycin (rhizoxin); azofurans (sizofurans); germanium spiroamines (spirogyranium); tenuazonic acid (tenuazonic acid); triimine quinone (triaziquone); 2, 2', 2 "-trichlorotriethylamine; trichothecene toxins (particularly T-2 toxin, myxotoxin a (verracutin a), bacillus toxin (roridin a) and snake toxin (anguidine)); urethane (urethan); vindesine (vindesine); dacarbazine (dacarbazine); mannitol mustard (mannomustine); dibromomannitol (mitobronitol); dibromodulcitol (mitolactol); pipobromane (pipobroman); guaxitucine (gamytosine); cytarabine ("Ara-C"); cyclophosphamide (cyclophosphamide); thiotepa (thiotepa); taxanes, e.g. TAXOL^TMPaclitaxel (Bristol-Myers Squibb Oncology, Princeton, N.J.), ABRAXANE^TMCremophor-free, albumin engineered nanoparticle paclitaxel formulations (American Pharmaceutical Partners, Schaumberg, Ill.) and paclitaxel (TAXOTERE @)^TM) Nanoparticle formulations of docetaxel (doxetaxel) (Rhone-Poulenc Rorer, Antony, France); chlorambucil (chlorenbucil); GEMZAR^TMGemcitabine; 6-thioguanine; mercaptopurine; methotrexate; platinum analogs such as cisplatin, oxaliplatin, and carboplatin; vinblastine; platinum; etoposide (VP-16); ifosfamide; mitoxantrone; vincristine; navelbine (navelbine. rtm), vinorelbine; nuantro (novantrone); teniposide (teniposide); edatrexate (edatrexate); daunorubicin; aminopterin (aminopterin); (xiloda); ibandronate (ibandronate); irinotecan (irinotecan) (Camptosar, CPT-11) (treatment regimens involving irinotecan with 5-FU and folinic acid); topoisomerase inhibitor RFS 2000; difluoromethyl ornithine (DMFO); retinoids, such as retinoic acid; capecitabine; combretastatin (combretastatin); folinic acid (LV); oxaliplatin, including oxaliplatin treatment regimen (FOLFOX); lapatinib (Tykerb)^TM) PKC- α, Raf, H-Ras, EGFR (e.g., erlotinib (Tarceva) for reducing cell proliferation^TM) And VEGF-a, and pharmaceutically acceptable salts, acids, or derivatives of any of the foregoing.

The definition also includes anti-hormonal agents used to modulate or inhibit hormonal effects on tumors, such as anti-estrogens and Selective Estrogen Receptor Modulators (SERMs), including, for example, tamoxifen (tamoxifen) (including NOLVADEX)^TMTamoxifen), raloxifene (raloxifene), droloxifene (droloxifene), 4-hydroxytamoxifene, trioxifene (trioxifene), raloxifene (keoxifene), LY117018, onapristone (onapristone), and FARESTON toremifene (toremifene); aromatase inhibitors which inhibit aromatase, which regulates estrogen production in the adrenal gland, e.g., 4(5) -imidazole, aminoglutethimide, MEGASE^TMMegestrol acetate, AROMASIN^TMExemestane, formestane (formestanine), fadrozole and rivoror^TM，FEMARA^TMLetrozole and ARIMIDEX^TMAnastrozole, and antiandrogens such as flutamide, nilutamide, bicalutamide, leuprorelin and goserelin, and troxacitabine (1, 3-dioxolane nucleoside cytosine analogues), antisense oligonucleotides, particularly those that inhibit expression of genes in signaling pathways associated with abnormal cell proliferation, such as PKC- α, Ralf and H-ras, ribozymes, such as VEGF expression inhibitors (e.g., ANGIOZYME)^TMRibozymes) and inhibitors of HER2 expression; vaccines, e.g. gene therapy vaccines, e.g. ALLOVECTIN^TMVaccine, LEUVECTIN^TMVaccine and VAXID^TMA vaccine; PROLEUKIN^TMrIL-2；LURTOTECAN^TMA topoisomerase 1 inhibitor; ABARELIX^TMrmRH; and pharmaceutically acceptable salts, acids or derivatives of any of the foregoing.

In some embodiments, the chemotherapeutic agent is selected from: actinomycin, Alitretinoin (Alitretinoin), all-trans retinoic acid, azacitidine, azathioprine, bevacizumab, bexarotene (bexarotene), bleomycin, bortezomib, carboplatin, capecitabine, cetuximab, cisplatin, chlorambucil, cyclophosphamide, cytarabine, daunorubicin, docetaxel, doxifluridine, doxorubicin, epirubicin, epothilone, etoposide, fluorouracil, gefitinib, gemcitabine, hydroxyurea, idarubicin, imatinib, easily pimameb (Ipilimumab), irinotecan, mechlorethamine, melphalan, mercaptopurine, methotrexate, mitoxantrone, omab, Ofatumumab (Ofatumumab), oxaliplatin, paclitaxel, panitumumab (Panitumab), memantin, rituximab, tafenoside (tafiniposiposide), topotecan (topotecan), topotecan, oxaliplatin, paclitaxel (Panitumab), mellitorimab, rituximab, and tebuflomezine (tazapine), Teniposide (Teniposide), topotecan (tebuflomentab), and a, Tretinoin, valrubicin, Vemurafenib, vinblastine, vincristine, vindesine, vinorelbine, vorinostat, romidepsin, 5-fluorouracil (5-FU), 6-mercaptopurine (6-MP), cladribine, clofarabine, floxuridine, fludarabine, pentostatin, mitomycin, ixabepilone, estramustine, prednisone, methylprednisolone, dexamethasone, or combinations thereof.

The term "cytokine" is a generic term for proteins released by one cell population as intercellular mediators to act on another cell. Examples of such cytokines are lymphokines, monokines, and traditional polypeptide hormones. The cell factor includes growth hormone, such as human growth hormone, N-methionyl human growth hormone and bovine growth hormone; parathyroid hormone; thyroxine; insulin; proinsulin; relaxin; (ii) prorelaxin; glycoprotein hormones such as Follicle Stimulating Hormone (FSH), Thyroid Stimulating Hormone (TSH) and Luteinizing Hormone (LH); an epidermal growth factor; a liver growth factor; fibroblast growth factor; prolactin; placental lactogen; tumor necrosis factor-alpha and-beta; (ii) a muller's inhibitor; a murine gonadotropin-related peptide; a statin; an activin; vascular endothelial growth factor; an integrin; thrombopoietin (TPO); nerve growth factors, such as NGF-alpha; platelet growth factor; transforming Growth Factors (TGF), such as TGF-alpha and TGF-beta; insulin-like growth factors-I and-II; erythropoietin (EPO); (ii) an osteoinductive factor; interferons, such as interferon- α, - β and- λ; colony Stimulating Factors (CSFs), such as macrophage-CSF (M-CSF); granulocyte-macrophage-CSF (GM-CSF); and granulocyte-CSF (G-CSF); interleukins (IL), such as IL-1, IL-1 α, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-11, IL-12; tumor necrosis factors, such as TNF- α or TNF- β; and other polypeptide factors, including LIF and Kit Ligand (KL). As used herein, the term cytokine includes proteins from natural sources or from recombinant cell culture as well as biologically active equivalents of the native sequence cytokines.

"growth inhibitory agent" as used herein refers to a compound or composition that inhibits cell growth in vitro and/or in vivo. Thus, growth inhibitors can significantly reduce the percentage of cells in S phase. Examples of growth inhibitory agents include agents that block cell cycle progression (at a position outside the S phase), such as agents that induce G1 arrest and M phase arrest. Typical M-phase blockers include vinca rosea (vincristine and vinblastine), TAXOL^TMAnd topo II inhibitors such as doxorubicin, epirubicin, daunorubicin, etoposide, and bleomycin. Those agents that arrest G1 also extend to S phase blockade, for example, DNA alkylating agents such as tamoxifen, prednisone, dacarbazine, mechlorethamine, cisplatin, methotrexate, 5-fluorouracil, and cytarabine. More information can be found in: the Molecular Basis of Cancer, edited by Mendelsohn and Israel, Chapter 1, entitled "cellulose regulation, oncogenes, and anti-cosmetic drugs", Murakami et al (WB Saunders: Philadelphia, 1995), essentially at page 13.

"radiation therapy" refers to the use of directed gamma or beta radiation to induce sufficient damage to cells to limit their ability to function properly or to destroy cells completely. It will be appreciated that there are many methods known in the art to determine the dose and duration of treatment. Typical treatments are given in a single administration and a typical dose range of 10 to 200 units (gray) per day.

The term "statistically significant" or "significant" means that the statistical evidence is discriminative. Defined as the probability that a decision is made to exclude a zero hypothesis when it is actually true. This decision is often made using p-values.

The term "functional" when used in conjunction with an "equivalent," "analog," "derivative," or "variant" or "fragment" refers to an entity or molecule having a biological activity substantially similar to the biological activity of the entity or molecule that is the equivalent, analog, derivative, variant, or fragment thereof.

As used herein, the term "aliphatic" refers to moieties characterized by a linear or branched arrangement of constituent carbon atoms, which may be saturated or partially unsaturated with one or more (e.g., one, two, three, four, five or more) double or triple bonds.

As used herein, the term "cycloaliphatic" refers to a moiety that includes a non-aromatic ring structure. The alicyclic moiety may be saturated or partially unsaturated with one or more double or triple bonds. The cycloaliphatic moiety may also optionally contain heteroatoms such as nitrogen, oxygen and sulfur. The nitrogen atoms may optionally be quaternized or oxidized, and the sulfur atoms may optionally be oxidized. Examples of alicyclic moieties include, but are not limited to, those having C₃-C₈Cyclic moieties such as cyclopropyl, cyclohexane, cyclopentane, cyclopentene, cyclopentadiene, cyclohexane, cyclohexene, cyclohexadiene, cycloheptane, cycloheptene, cycloheptadiene, cyclooctane, cyclooctene and cyclooctadiene.

As used herein, the term "alkyl" refers to a straight or branched chain saturated aliphatic group having a chain of carbon atoms. Usually using C_xAlkyl and C_x-C_yAlkyl, wherein X and Y indicate the number of carbon atoms in the chain. E.g. C₁-C₆Alkyl groups include alkyl groups having chains of 1 to 6 carbons (e.g., methyl, ethyl, propyl, isopropyl, butyl, sec-butyl, isobutyl, tert-butyl, pentyl, neopentyl, hexyl, and the like). Alkyl (e.g., arylalkyl) represented with another group refers to a straight or branched chain saturated alkyl group having the indicated number of atomsDivalent radicals, or when no atoms are indicated, meaning bonds, e.g. (C)₆-C₁₀) Aryl radical (C)₀-C₃) Alkyl groups include phenyl, benzyl, phenethyl, 1-phenylethyl, 3-phenylpropyl, and the like. The backbone of the alkyl group may optionally be interrupted by one or more heteroatoms, such as N, O or S. The term "alkyl" includes heteroalkyl.

In some preferred embodiments, the linear or branched alkyl group has 30 or fewer carbon atoms in its backbone (e.g., C1-C30 linear, C3-C30 branched), more preferably 20 or fewer. Likewise, preferred cycloalkyl groups have 3 to 10 carbon atoms in their ring structure, and more preferably have 5, 6 or 7 carbons in the ring structure. The term "alkyl" (or "lower alkyl") as used throughout the specification, examples and claims is intended to include both "unsubstituted alkyls" and "substituted alkyls," the latter of which refers to alkyl moieties having one or more substituents replacing a hydrogen on one or more carbons of the hydrocarbon backbone.

As used herein, "lower alkyl" refers to an alkyl group as defined above, but having from one to ten carbons in its backbone structure, more preferably one to six carbon atoms, unless the number of carbons is otherwise specified. Likewise, "lower alkenyl" and "lower alkynyl" have similar chain lengths. Throughout this application, preferred alkyl groups are lower alkyl groups.

In some preferred embodiments, the substituents designated herein as alkyl are lower alkyl.

Substituents for substituted alkyl groups may include halogen, hydroxy, nitro, thiol, amino, azido, imino, amido, phosphoryl (including phosphonic and phosphinic acids), sulfonyl (including sulfuric, sulfonamido, sulfamoyl and sulfonic acids) and silyl groups, as well as ether, alkylthio, carbonyl (including ketones, aldehydes, carboxylic acids and esters), -CF3, -CN, and the like.

As used herein, the term "alkenyl" refers to an unsaturated straight, branched, or cyclic hydrocarbon group having at least one carbon-carbon double bond. Usually using C_xAlkenyl and C_x-C_yAlkenyl, wherein X and Y indicate the number of carbon atoms in the chain. E.g. C₂-C₆Alkenyl groups include alkenyl groups having a chain of 1 to 6 carbons and at least one double bond, such as vinyl, allyl, propenyl, isopropenyl, 1-butenyl, 2-butenyl, 3-butenyl, 2-methylallyl, 1-hexenyl, 2-hexenyl, 3-hexenyl, and the like). Alkenyl groups represented with another group (e.g., such as arylalkenyl) refer to straight or branched chain alkenyl divalent groups having the indicated number of atoms. The backbone of the alkenyl group may optionally be interrupted by one or more heteroatoms, such as N, O or S.

As used herein, the term "alkynyl" refers to an unsaturated hydrocarbon group having at least one carbon-carbon triple bond. Usually using C_xAlkynyl and C_x-C_yAlkynyl, wherein X and Y indicate the number of carbon atoms in the chain. E.g. C₂-C₆Alkynyl includes alkynyl groups having a chain of 1 to 6 carbons and at least one triple bond, such as ethynyl, 1-propynyl, 2-propynyl, 1-butynyl, isopentynyl, 1, 3-hex-diynyl-yl, n-hexynyl, 3-pentynyl, 1-hex-3-ynyl, and the like. Alkynyl (e.g., as arylalkynyl) represented with another group refers to a straight or branched chain alkynyl divalent group having the indicated number of atoms. The backbone of the alkynyl group may optionally be interrupted by one or more heteroatoms, such as N, O or S.

The terms "alkylene," "alkenylene," and "alkynylene" refer to divalent alkyl, alkenylene, and alkynylene groups. Usually using the prefix C_xAnd C_x-C_yWherein X and Y indicate the number of carbon atoms in the chain. E.g. C₁-C₆Alkylene groups include methylene (-CH)₂-) ethylene (-CH₂CH₂-) propylene (-CH)₂CH₂CH₂-) tetramethylene group (-CH₂CH₂CH₂CH₂-), 2-methyltetramethylene (-CH)₂CH(CH₃)CH₂CH₂-) pentamethylene (-CH₂CH₂CH₂CH₂CH₂-) etc.).

As used herein, the term "alkylene" refers to a straight or branched chain unsaturated aliphatic divalent group having the general formula ═ CR_aR_b. Usually using C_xAlkylene and C_x-C_yAlkylene, wherein X and Y indicate the number of carbon atoms in the chain. E.g. C₂-C₆Alkylene includes methylene (═ CH)₂) Ethylene (═ CHCH)₃) Isopropylidene (═ C (CH)₃)₂) Propylene (═ CHCH)₂CH₃) Allyl idene (═ CH-CH ═ CH)₂) Etc.).

As used herein, the term "heteroalkyl" refers to a straight-chain or branched-chain or cyclic carbon-containing group, or a combination thereof, that contains at least one heteroatom. Suitable heteroatoms include, but are not limited to, O, N, Si, P, Se, B, and S, wherein the phosphorus and sulfur atoms are optionally oxidized, and the nitrogen heteroatom may be optionally quaternized. The heteroalkyl group may be substituted as defined above for alkyl.

As used herein, the term "halogen" or "halo" refers to an atom selected from fluorine, chlorine, bromine, and iodine. The term "halogen radioisotope" or "halogen isotope" refers to a radionuclide of an atom selected from fluorine, chlorine, bromine and iodine.

A "halogen-substituted moiety" or "halo moiety" as a separate group or part of a larger group means an aliphatic, alicyclic, or aromatic moiety described herein substituted with one or more "halogen" atoms (as such terms are defined herein). For example, halo-substituted alkyl includes haloalkyl, dihaloalkyl, trihaloalkyl, perhaloalkyl, and the like (e.g., halo (C)₁-C₃) The alkyl group includes chloromethyl, dichloromethyl, difluoromethyl, trifluoromethyl (-CF)₃) 2, 2, 2-trifluoroethyl, perfluoroethyl, 2, 2, 2-trifluoro-1, 1-dichloroethyl, etc.).

The term "aryl" refers to a monocyclic, bicyclic, or tricyclic fused aromatic ring system. Usually using C_xAryl and C_x-C_yAryl, in which X and Y denote in the ring systemThe number of carbon atoms. Exemplary aryl groups include, but are not limited to, pyridyl, pyrimidinyl, furyl, thienyl, imidazolyl, thiazolyl, pyrazolyl, pyridazinyl, pyrazinyl, triazinyl, tetrazolyl, indolyl, benzyl, phenyl, naphthyl, anthracyl, thienyl, fluorenyl, indanyl, indenyl, naphthyl, phenyl, tetrahydronaphthyl, benzimidazolyl, benzofuranyl, benzothiofuranyl, benzothiophenyl, benzoxazolyl, benzoxazolinyl, benzothiazolyl, benzotriazolyl, benzotetrazolyl, benzisoxazolyl, benzisothiazole, benzimidazolinyl, carbazolyl, 4aH carbazolyl, carbolinyl, chromanyl, chromenyl, cinnolinyl, decahydroquinolinyl, 2H, 6H-1, 5, 2-dithiazinyl, dihydrofuro [2, 3b ] furoyl]Tetrahydrofuran, furyl, furazanyl, imidazolidinyl, imidazolinyl, imidazolyl, 1H-indazolyl, indolinyl, indolizinyl, indolyl, 3H-indolyl, isatoiyl, isobenzofuryl, isochromanyl, isoindolyl, isoindoline, isoindolyl, isoquinolyl, isothiazolyl, isoxazolyl, methylenedioxyphenyl, morpholinyl, naphthyridinyl, octahydroisoquinolyl, oxadiazolyl, 1, 2, 3-oxadiazolyl, 1, 2, 4-oxadiazolyl, 1, 2, 5-oxadiazolyl, 1, 3, 4-oxadiazolyl, oxazolidinyl, oxazolyl, oxindolyl, pyrimidinyl, phenanthridinyl, phenanthrolinyl, phenazinyl, phenothiazinyl, phenoxathiinyl, phenazinyl, phthalazinyl, piperazinyl, piperidinyl, piperidine, piperidyl, quinoxalinyl, phenanthrolinyl, phthalazinyl, piperazinyl, piperidyl, and pyridonyl, 4-piperidonyl, piperonyl, pteridinyl, purinyl, pyranyl, pyrazinyl, pyrazolidinyl, pyrazolinyl, pyrazolyl, pyridazinyl, pyridooxazolyl (pyridooxazole), pyridoimidazolyl, pyridothiazolyl, pyridyl (pyridinyl), pyridyl (pyridyll), pyrimidinyl, pyrrolidinyl, pyrrolinyl, 2H-pyrrolyl, quinazolinyl, quinolinyl, 4H-quinolizinyl, quinoxalinyl, quinuclidinyl, tetrahydrofuranyl, tetrahydroisoquinolinyl, tetrahydroquinolinyl, tetrazolyl, 6H-1, 2, 5-thiadiazinyl, 1, 2, 3-thiadiazolyl, 1, 2, 4-thiadiazolyl, 1, 2, 5-thiadiazolyl, 1, 3, 4-thiadiazolyl, thianthrenyl, thiazolyl, thienyl, thiazolothiazolylOxazolyl, thiazoloimidazolyl, thienyl, xanthenyl, and the like. In some embodiments, 1, 2, 3, or 4 hydrogen atoms of each ring may be substituted with a substituent.

The term "heteroaryl" refers to an aromatic 5-8 membered monocyclic, 8-12 membered fused bicyclic, or 11-14 membered fused tricyclic ring system having 1-3 heteroatoms (if monocyclic), 1-6 heteroatoms (if bicyclic), or 1-9 heteroatoms (if tricyclic) selected from O, N or S (e.g., carbon atoms and 1-3, 1-6, or 1-9N, O or S heteroatoms, if monocyclic, bicyclic, or tricyclic, respectively). Usually using C_xHeteroaryl and C_x-C_yHeteroaryl, wherein X and Y indicate the number of carbon atoms in the ring system. Heteroaryl groups include, but are not limited to, those derived from: benzo [ b ]]Furan, benzo [ b ]]Thiophene, benzimidazole, imidazo [4, 5-c ]]Pyridine, quinazoline, thieno [2, 3-c ]]Pyridine, thieno [3, 2-b ]]Pyridine, thieno [2, 3-b ]]Pyridine, indolizine, imidazo [1, 2a ]]Pyridine, quinoline, isoquinoline, phthalazine, quinoxaline, naphthyridine, quinolizine, indole, isoindole, indazole, indoline, benzoxazole, benzopyrazole, benzothiazole, imidazo [1, 5-a ]]Pyridine, pyrazolo [1, 5-a ]]Pyridine, imidazo [1, 2-a ]]Pyrimidine, imidazo [1, 2-c ]]Pyrimidine, imidazo [1, 5-a ]]Pyrimidine, imidazo [1, 5-c ]]Pyrimidine, pyrrolo [2, 3-b ]]Pyridine, pyrrolo [2, 3c]Pyridine, pyrrolo [3, 2-c]Pyridine, pyrrolo [3, 2-b ]]Pyridine, pyrrolo [2, 3-d ] s]Pyrimidine, pyrrolo [3, 2-d]Pyrimidine, pyrrolo [2, 3-b ]]Pyrazine, pyrazolo [1, 5-a ]]Pyridine, pyrrolo [1, 2-b ]]Pyridazine, pyrrolo [1, 2-c ]]Pyrimidine, pyrrolo [1, 2-a ]]Pyrimidine, pyrrolo [1, 2-a ]]Pyrazine, triazolo [1, 5-a ]]Pyridine, pteridine, purine, carbazole, acridine, phenazine, phenothiazine, phenothizine, 1, 2-dihydropyrrolo [3, 2, 1-hi]Indole, indolizine, pyrido [1, 2-a ]]Indole, 2(1H) -pyridone, benzimidazolyl, benzofuranyl, benzothiofuranyl, benzothiophenyl, benzoxazolyl, benzoxazolinyl, benzothiazolyl, benzotriazolyl, benzotetrazolyl, benzisoxazolyl, benzisothiazole, benzimidazolinyl, carbazolyl, 4aHCarbazolyl, carbolinyl, chromanyl, chromenyl, cinnolinyl, decahydroquinolinyl, 2H, 6H-1, 5, 2-diazinyl, dihydrofuro [2, 3-b ]]Tetrahydrofuran, furyl, furazanyl, imidazolidinyl, imidazolinyl, imidazolyl, 1H-indazolyl, indolinyl, indolizinyl, indolyl, 3H-indolyl, isatoiyl, isobenzofuran, isochroman, isoindolyl, isoquinolinyl, isothiazolyl, isoxazolyl, methylenedioxyphenyl, morpholinyl, naphthyridinyl, octahydroisoquinolinyl, oxadiazolyl, 1, 2, 3-oxadiazolyl, 1, 2, 4-oxadiazolyl, 1, 2, 5-oxadiazolyl, 1, 3, 4-oxadiazolyl, oxazolidinyl, oxazolyl, oxepanyl, oxetanyl, oxindolyl, pyrimidinyl, phenanthridinyl, phenanthrolinyl, phenazinyl, phenothiazinyl, phenoxathiinyl, phenazinyl, piperazinyl, isothiazolyl, indolyl, indolinyl, octahydroisoquinolyl, isothiazolyl, 1, 2, 3, 4-oxadiazolyl, 1, 3, 4-oxadiazolyl, oxazolidinyl, oxazolyl, oxepanyl, Piperidinyl, piperidonyl, 4-piperidonyl, piperonyl, pteridinyl, purinyl, pyranyl, pyrazinyl, pyrazolidinyl, pyrazolinyl, pyrazolyl, pyridazinyl, pyridooxazolyl, pyridoimidazolyl, pyridothiazolyl, pyridinyl, pyrimidinyl, pyrrolidinyl, pyrrolinyl, 2H-pyrrolyl, quinazolinyl, quinolinyl, 4H-quinolizinyl, quinoxalinyl, quinuclidinyl, tetrahydrofuranyl, tetrahydroisoquinolinyl, tetrahydropyranyl, tetrahydroquinolinyl, tetrazolyl, 6H-1, 2, 5-thiadiazinyl, 1, 2, 3-thiadiazolyl, 1, 2, 4-thiadiazolyl, 1, 2, 5-thiadiazolyl, 1, 3, 4-thiadiazolyl, thianthrenyl, thiazolyl, thienyl, thiazolo, thiazolyl, thiazolo, thiazolyl, Thiazolooxazolyl, thiazoloimidazolyl, thienyl, and xanthyl groups. Some exemplary heteroaryl groups include, but are not limited to, pyridyl, furyl (furyl or furanyl), imidazolyl, benzimidazolyl, pyrimidinyl, thienyl (thiophenyl or thienyl), pyridazinyl, pyrazinyl, quinolinyl, indolyl, thiazolyl, naphthyridinyl, 2-amino-4-oxo-3, 4-dihydropteridin-6-yl, tetrahydroisoquinoline, and the like. In some embodiments, 1, 2, 3, or 4 hydrogen atoms of each ring may be substituted with a substituent.

The term "cyclyl" or "cycloalkyl" refers to a cyclic group having 3 to 12 atomsCarbon, for example, saturated and partially unsaturated cyclic hydrocarbon groups of 3 to 8 carbons, for example 3 to 6 carbons. Usually using C_xCyclic group and C_x-C_yCyclic group, wherein X and Y indicate the number of carbon atoms in the ring system. Cycloalkyl groups may also be optionally substituted, for example, with 1, 2, 3, or 4 substituents. C₃-C₁₀The cyclic group includes cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclohexenyl, 2, 5-cyclohexadienyl, cycloheptyl, cyclooctyl, bicyclo [2.2.2]Octyl, adamantan-1-yl, decahydronaphthyl, oxocyclohexyl, dioxocyclohexyl, thioxocyclohexyl, 2-oxobicyclo [2.2.1]Hept-1-yl, and the like.

Aryl and heteroaryl groups may be optionally substituted at one or more positions with one or more substituents, for example, halogen, alkyl, aralkyl, alkenyl, alkynyl, cycloalkyl, hydroxy, amino, nitro, mercapto, imino, amido, phosphate, phosphonate, phosphinate, carbonyl, carboxyl, silyl, ether, alkylthio, sulfonyl, ketone, aldehyde, ester, heterocyclyl, aromatic or heteroaromatic moieties, -CF3, -CN, and the like.

The term "heterocyclyl" refers to a non-aromatic 5-8 membered monocyclic, 8-12 membered bicyclic, or 11-14 membered tricyclic ring system having 1-3 heteroatoms (if monocyclic), 1-6 heteroatoms (if bicyclic), or 1-9 heteroatoms (if tricyclic) selected from O, N or S (e.g., carbon atoms and 1-3, 1-6, or 1-9 heteroatoms N, O or S, respectively, if monocyclic, bicyclic, or tricyclic). Usually using C_xHeterocyclyl and C_x-C_yHeterocyclyl, wherein X and Y indicate the number of carbon atoms in the ring system. In some embodiments, 1, 2, or 3 hydrogen atoms of each ring may be substituted with a substituent. Exemplary heterocyclic groups include, but are not limited to, piperazinyl, pyrrolidinyl, dioxanyl, morpholinyl, tetrahydrofuranyl, piperidinyl, 4-morpholine, 4-piperazinyl, pyrrolidinyl, perhydropyrrolidinyl (perhydropyrrolizinyl), 1, 4-diazaperhydrocycloheptyl (1, 4-diazaperhydropyridinyl), 1, 3-dioxanyl, 1, 4-dioxanyl, and the like.

The terms "bicyclic" and "tricyclic" refer to a collection of multiple rings that are fused, bridged, or bound by single bonds.

The term "cycloalkylene" refers to a divalent aryl, heteroaryl, cyclyl, or heterocyclyl group.

As used herein, the term "fused ring" refers to a ring that is bonded to another ring to form a compound having a bicyclic structure, when the ring atoms common to both rings are directly bonded to each other. Non-exclusive examples of common fused rings include decalin, naphthalene, anthracene, phenanthrene, indole, furan, benzofuran, quinoline, and the like. The compounds having a fused ring system may be saturated, partially saturated cyclic groups, heterocyclic groups, aromatic hydrocarbons, heteroaromatic hydrocarbons, and the like.

As used herein, the term "carbonyl" refers to the group-C (O) -. It is noted that the carbonyl group can be further substituted with a variety of substituents to form different carbonyl groups including acids, acid halides, amides, esters, ketones, and the like.

The term "carboxy" refers to the group-C (O) O-. Notably, the compounds described herein as containing a carboxyl moiety may include protected derivatives thereof, i.e., wherein the oxygen is substituted with a protecting group. Suitable protecting groups for the carboxy moiety include benzyl, t-butyl, and the like. The term "carboxyl" means-COOH

The term "cyano" refers to the group-CN.

The term "heteroatom" refers to an atom that is not a carbon atom. Specific examples of heteroatoms include, but are not limited to, nitrogen, oxygen, sulfur, and halogens. "heteroatom moiety" includes moieties wherein the atom to which the moiety is attached is not carbon. Examples of heteroatom moieties include-N ═ -NR^N-、-N⁺(O^-) -O-, -S-or-S (O)₂-、-OS(O)₂-and-SS-, wherein R^NIs H or another substituent.

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

The term "imine derivative" refers to derivatives containing the moiety-C (NR) -wherein R comprises a hydrogen or carbon atom alpha to the nitrogen.

The term "nitro" refers to the group-NO₂。

"oxaaliphatic", "oxacycloaliphatic" or "oxaaromatic" refers to aliphatic, cycloaliphatic or aromatic as defined herein, except that one or more oxygen atoms (-O-) are positioned between aliphatic, cycloaliphatic or aromatic carbon atoms.

"oxoaliphatic", "oxoalicyclic" or "oxoaromatic" refers to aliphatic, alicyclic or aromatic groups as defined herein substituted with a carbonyl group. The carbonyl group can be an aldehyde, ketone, ester, amide, acid, or acid halide.

As used herein, the term "aromatic" refers to a moiety wherein the constituent atoms form an unsaturated ring system, all of the atoms in the ring system being sp²Hetero and the total number of pi electrons is equal to 4n + 2. The aromatic ring may be such that the ring atoms are only carbon atoms (e.g., aryl), or may include carbon and non-carbon atoms (e.g., heteroaryl).

As used herein, the term "substituted" refers to the independent replacement of one or more (typically 1, 2, 3, 4, or 5) hydrogen atoms on the substituted moiety with substituents independently selected from the group of substituents listed or otherwise indicated below in the definition of "substituent". In general, a non-hydrogen substituent may be any substituent capable of binding to the atom of a given moiety designated as substituted. Examples of substituents include, but are not limited to, acyl, amido, acyloxy, aldehyde, cycloaliphatic, aliphatic, alkanesulfonamide, alkanesulfonyl, alkaryl, alkenyl, alkoxy, alkoxycarbonyl, alkyl, alkylamino, alkylcarbamoyl, alkylene, alkylidene, alkylthio, alkynyl, amido, amino, aminoalkyl, aralkyl, aralkylsulfonamido, arylsulfonyl, aromatic, aryl, arylamino, arylcarbamoyl, aryloxy, azido, carbamoyl, carbonyl, and the like,Carbonyls (including keto, carboxyl, carboxylate, CF)₃Cyano (CN), cycloalkyl, cycloalkylene, ester, ether, haloalkyl, halogen, heteroaryl, heterocyclyl, hydroxy, hydroxyalkyl, imino, iminoketone, ketone, mercapto, nitro, oxaalkyl, oxo, oxoalkyl, phosphoryl (including phosphonic and phosphinic acids), silyl, sulfonamide, sulfonyl (including sulfate, sulfamoyl, and sulfonate), thiol, and ureido moieties, each of which may also be optionally substituted or unsubstituted. In some cases, two substituents together with the carbon to which they are attached may form a ring.

The term "alkoxy" or "alkoxy" as used herein refers to an alkyl, alkenyl, alkynyl, aryl, cyclyl or heterocyclyl group as defined above having an oxygen radical attached thereto. Representative alkoxy groups include methoxy, ethoxy, propoxy, t-butoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, and the like. An "ether" is two hydrocarbons covalently linked by oxygen. Thus, an alkyl substituent that renders an alkyl group as an ether is or is analogous to an alkoxy group, and may be represented, for example, by one of: -O-alkyl, -O-alkenyl, -O-alkynyl, -O-cyclyl, -O-heterocyclyl, -O-aryl and-O-heteroaryl. The term "alkoxy" includes aryloxy (aroxy and aryloxy). Aryloxy groups may be represented by-O-aryl or O-heteroaryl groups, wherein aryl and heteroaryl are defined below. Alkoxy and aryloxy groups may be substituted as described above for alkyl groups.

As used herein, the term "aralkyl" refers to an alkyl group substituted with an aryl group (e.g., an aromatic or heteroaromatic group).

The term "alkylthio" refers to an alkyl group as defined above having a sulfur radical attached thereto. In some preferred embodiments, an "alkylthio" moiety is represented by one of-S-alkyl, -S-alkenyl, and-S-alkynyl. Representative alkylthio groups include methylthio, ethylthio, and the like. The term "alkylthio" also includes cycloalkyl, alkene and cycloalkene radicals and alkynyl radicals. The term "alkylthio" also includes arylthio. "arylthio" refers to aryl or heteroaryl.

The term "sulfinyl" refers to the radical-SO-. It is noted that the sulfinyl group can be further substituted with a variety of substituents to form different sulfinyl groups, including sulfinic acids, sulfinamides, sulfinyl esters, sulfoxides, and the like.

The term "sulfonyl" refers to the group-SO₂-. It is noted that the sulfonyl group can be further substituted with a variety of substituents to form various sulfonyl groups, including sulfonic acids (-SO)₃H) Sulfonamides, sulfonates, sulfones, and the like.

The term "thiocarbonyl" refers to the group-C (S) -. It is noted that thiocarbonyl groups may be further substituted with a variety of substituents to form different thiocarbonyl groups including thioacids, thioamides, thioesters, thioketones and the like.

As used herein, the term "amino" refers to-NH₂. The term "alkylamino" refers to a nitrogen moiety having at least one straight or branched chain unsaturated aliphatic, cyclic, or heterocyclic group attached to the nitrogen. For example, representative amino groups include-NH₂、-NHCH₃、-N(CH₃)₂、-NH(C₁-C₁₀Alkyl), -N (C)₁-C₁₀Alkyl radical)₂And the like. The term "alkylamino" includes "alkenylamino", "alkynylamino", "cyclylamino" and "heterocyclylamino". The term "arylamino" refers to a nitrogen moiety having at least one aryl group attached to the nitrogen. For example-NH aryl and-N (aryl)₂And the like. The term "heteroarylamino" refers to a nitrogen moiety having at least one heteroaryl group attached to the nitrogen. For example-NH heteroaryl and-N (heteroaryl)₂And the like. Optionally, two substituents together with the nitrogen may also form a ring. Unless otherwise indicated, the amino moiety-containing compounds described herein may include protected derivatives thereof. Suitable protecting groups for the amino moiety include acetyl, t-butyloxycarbonyl, benzyloxycarbonyl and the like.

The term "aminoalkyl" refers to alkyl, alkenyl, and alkynyl groups as defined aboveExcept that one or more substituted or unsubstituted nitrogen atoms (-N-) are located between carbon atoms of the alkyl, alkenyl or alkynyl group. For example, (C)₂-C₆) Aminoalkyl refers to a chain containing from 2 to 6 carbons, and one or more nitrogen atoms between the carbon atoms.

The term "alkoxyalkoxy" refers to-O- (alkyl), such as-OCH₂CH₂OCH₃And the like.

The term "alkoxycarbonyl" refers to-C (O) O- (alkyl), such as-C (═ O) OCH₃、-C(＝O)OCH₂CH₃And the like.

The term "alkoxyalkyl" refers to- (alkyl) -O- (alkyl), such as-CH₂OCH₃、-CH₂OCH₂CH₃And the like.

The term "aryloxy" refers to-O- (aryl), such as-O-phenyl, -O-pyridyl, and the like.

The term "aralkyl" refers to- (alkyl) - (aryl) or- (alkyl) - (heteroaryl), such as benzyl (i.e., -CH)₂Phenyl), -CH₂-pyridyl and the like.

The term "arylalkoxy" refers to-O- (alkyl) - (aryl) or-O- (alkyl) - (heteroaryl), such as-O-benzyl, -O-CH₂-pyridyl and the like.

The term "cycloalkoxy" refers to-O- (cycloalkyl), such as-O-cyclohexyl, and the like.

The term "cycloalkylalkoxy" refers to-O- (alkyl) - (cycloalkyl), such as-OCH₂Cyclohexyl, and the like.

The term "aminoalkoxy" refers to-O- (alkyl) -NH₂E.g. -OCH₂NH₂、-OCH₂CH₂NH₂And the like.

The term "mono-or di-alkylamino" refers to-NH (alkyl) or-N (alkyl), such as-NHCH, respectively₃、-N(CH₃)₂And the like.

The term "mono-or di-alkylaminoalkoxy" means-O- (alkyl) -NH (alkyl) or-O- (alkyl) -N (alkyl), such as-OCH₂NHCH₃、-OCH₂CH₂N(CH₃)₂And the like.

The term "arylamino" refers to-NH (aryl), such as-NH-phenyl, -NH-pyridyl, and the like.

The term "arylalkylamino" refers to-NH- (alkyl) - (aryl), such as-NH-benzyl, -NHCH₂-pyridyl and the like.

The term "alkylamino" refers to-NH (alkyl), such as-NHCH₃、-NHCH₂CH₃And the like.

The term "cycloalkylamino" refers to-NH- (cycloalkyl), such as-NH-cyclohexyl, and the like.

The term "cycloalkylalkylamino" -NH- (alkyl) - (cycloalkyl), such as-NHCH₂Cyclohexyl and the like.

With respect to all definitions provided herein, it is noted that these definitions should be construed as open-ended in the sense that substituents beyond those specified may be included. Thus, C₁Alkyl represents a substituent having one carbon atom, but does not represent a carbon atom. Thus, C₁Alkyl groups include methyl (i.e., -CH3) and-CR_aR_bR_cWherein R is_a、R_bAnd R_cMay each independently be hydrogen or any other substituent wherein the atom at carbon α is a heteroatom or cyano₃、CH₂OH and CH₂CN is all C₁An alkyl group.

The term "derivative" as used herein refers to a chemical substance that is structurally related to another chemical substance, the "original" substance, which may be referred to as the "parent" compound. A "derivative" may be made from the structurally related parent compound in one or more steps. In some embodiments, the general physical and chemical properties of the derivative may be similar or different from the parent compound.

Unless otherwise indicated, structures depicted herein are intended to include compounds that differ only by the presence of one or more isotopically enriched atoms. For example, having the structure of the invention, replacing a hydrogen atom by deuterium or tritium, or replacing a carbon atom by deuterium or tritium¹³C or¹⁴C-enriched carbon compounds are within the scope of the invention.

As used herein, "pharmaceutically acceptable salt" is intended to encompass any compound described herein used in the form of a salt thereof, particularly where the salt confers improved pharmacokinetic properties on the compound as compared to the free form of the compound or a different salt form of the compound. The pharmaceutically acceptable salt forms may also initially confer desirable pharmacokinetic properties on the compound that it did not have before, and may even positively influence the pharmacodynamic effect of the compound with respect to its therapeutic activity in vivo. An example of a pharmacokinetic property that can be favorably influenced is the way in which a chemical is transported across the cell membrane, which in turn can directly and positively influence the absorption, distribution, biotransformation and excretion of the compound. Although the route of administration of a pharmaceutical composition is important, and various anatomical, physiological, and pathological factors can severely affect bioavailability, the solubility of a compound is often dependent upon the particular salt form in which it is used. One skilled in the art will recognize that aqueous solutions of the compounds will provide the most rapid absorption of the compounds into the subject being treated, while lipid solutions and suspensions, as well as solid dosage forms, will result in slower absorption of the compounds.

Pharmaceutically acceptable salts include those derived from inorganic acids such as sulfuric acid, sulfamic acid, phosphoric acid, nitric acid, and the like; and those derived from organic acids such as acetic acid, propionic acid, succinic acid, glycolic acid, stearic acid, lactic acid, malic acid, tartaric acid, citric acid, ascorbic acid, palmitic acid, maleic acid, hydroxymaleic acid, phenylacetic acid, glutamic acid, benzoic acid, salicylic acid, sulfanilic acid, 2-acetoxybenzoic acid, fumaric acid, toluenesulfonic acid, methanesulfonic acid, ethanedisulfonic acid, oxalic acid, isothiohydroxy acid (isothionic acid), and the like. See, e.g., Berge et al, "Pharmaceutical Salts", j.pharm.sci.66: 1-19(1977), the contents of which are herein incorporated by reference in their entirety. Exemplary salts also include hydrobromide, hydrochloride, sulfate, bisulfate, phosphate, nitrate, acetate, succinate, valerate, oleate, palmitate, stearate, laurate, benzoate, lactate, phosphate, tosylate, citrate, maleate, fumarate, succinate, tartrate, naphthoate, mesylate, glucoheptonate, lactobionate, laurylsulfonate, and the like. Suitable acids capable of forming salts with the compounds of the present disclosure include inorganic acids such as hydrochloric acid, hydrobromic acid, perchloric acid, nitric acid, thiocyanic acid, sulfuric acid, phosphoric acid, and the like; and organic acids such as 1, 2-ethanedisulfonic acid, 2-hydroxyethanesulfonic acid, 2-naphthalenesulfonic acid, 3-phenylpropionic acid, 4-methylbicyclo [2.2.2] oct-2-ene-1-carboxylic acid, 4' -methylene (3-hydroxy-2-ene-1-carboxylic acid), acetic acid, anthranilic acid, benzenesulfonic acid, benzoic acid, camphorsulfonic acid, cinnamic acid, citric acid, cyclopentanepropionic acid, ethanesulfonic acid, formic acid, fumaric acid, glucoheptonic acid, gluconic acid, glutamic acid, glycolic acid, heptanoic acid, hydroxynaphthoic acid, lactic acid, laurylsulfuric acid, maleic acid, malic acid, malonic acid, mandelic acid, methanesulfonic acid, muconic acid, naphthalenesulfonic acid, o- (4-hydroxybenzoyl) benzoic acid, oxalic acid, p-chlorobenzenesulfonic acid, propionic acid, p-toluenesulfonic acid, Pyruvic acid, salicylic acid, stearic acid, succinic acid, sulfanilic acid, tartaric acid, tert-butylacetic acid, trifluoroacetic acid, trimethylacetic acid, and the like. Suitable bases capable of forming salts with the compounds of the present disclosure include inorganic bases such as sodium hydroxide, ammonium hydroxide, sodium carbonate, calcium hydroxide, potassium hydroxide, and the like; and organic bases such as mono-, di-and tri-alkyl and aryl amines (e.g., triethylamine, diisopropylamine, methylamine, dimethylamine, N-methylglucamine, pyridine, picoline, dicyclohexylamine, N' -dibenzylethylenediamine, etc.), and optionally substituted ethanolamines (e.g., ethanolamine, diethanolamine, triethanolamine, etc.).

In some embodiments, the compounds described herein may be in the form of a prodrug. As used herein, the term "prodrug" refers to a compound that can be converted to a compound described herein by certain chemical or physiological processes (e.g., enzymatic processes and metabolic hydrolysis). Thus, the term "prodrug" also refers to pharmaceutically acceptable precursors of biologically active compounds. Upon administration to a subject, a prodrug may be inactive, i.e., an ester, but converted in vivo to the active compound, e.g., by hydrolysis to a free carboxylic acid or free hydroxyl group. Prodrug compounds generally offer the advantage of solubility in the organism, histocompatibility or delayed release. The term "prodrug" is also intended to include any covalently bonded carrier that releases the active compound in vivo when such a drug is administered to a subject. Prodrugs of an active compound as described herein may be prepared by modifying functional groups present in the active compound in such a way that the modification is cleaved, either in routine manipulation or in vivo, to the parent active compound. Prodrugs include compounds wherein a hydroxy, amino, or sulfhydryl group is bonded to any group that, when the prodrug of the active compound is administered to a subject, cleaves to form a free hydroxy, free amino, or free sulfhydryl group, respectively. For example, a compound comprising a hydroxyl group can be administered as an ester that is converted to a hydroxyl compound in vivo by hydrolysis. Suitable esters that can be converted in vivo to hydroxy compounds include acetate, citrate, lactate, tartrate, malonate, oxalate, salicylate, propionate, succinate, fumarate, formate, benzoate, maleate, methylene-bis-b-hydroxynaphthalene, cholate, isethionate, di-p-toluoyl tartrate, mesylate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate, cyclohexylsulfamate, quinic acid ester, esters of amino acids, and the like. Similarly, compounds containing amine groups can be administered as amides such as acetamides, formamides, and benzamides, which are converted to amine compounds in vivo by hydrolysis. See Harper, "Drug extension" edited in Jucker Progress in Drug Research 4: 221-294 (1962); morozowich et al, "Application of Physical Organic Principles to Prodrug Design" edited in E.B. Roche. Design of biological Properties through Prodrugs and domains, APHA Acad.pharm.Sci.40 (1977); bioreversible Carriers in Drug design, Theory and Application, e.b. roche editions, APHA acad. pharm. sci. (1987); designoff produgs, h.bundgaard, Elsevier (1985); wang et al, "Prodrug analogs to the improved delivery of peptide drivers" in curr. phase. design.5 (4): 265-287 (1999); pauletti et al (1997) Improvement in peptide bioavailability: peptidomimetics and Prodrug Strategies, adv. drug. delivery Rev.27: 235-256; mizen et al (1998) "The Use of drugs as precursors for organic Delivery of (3-latex anti-inflammatory," Drug. Biotech. ll.: 345; Gaignault et al (1996) "Delivery drugs and biological Delivery I.Carrier precursors," practice.Med.Chem.671-696; Asghannejad, "Imperial Oral Drug Delivery," Delivery drugs in pharmaceutical Delivery systems, G.L.Amidon, P.I.Lee and E.Marm.Tokyo edi, cell Delivery, p.185-218(2000) "," Banner et al, "The Use of drugs for Delivery drugs and Drug of Delivery drugs 183, 3. repair J.1997, 3. repair J.3. 1997, 3. biological Delivery J.1997-," filtration Drug of Delivery 1. J.3. 1997 "(1990) and" Delivery drugs for Delivery drugs of drugs and 3. repair. 3. biological Delivery J.3. repair. J., arch, pharm, chemi 86 (1): 1-39 (1979); "Improved drive Delivery by the Improved approach" and Controlled drive Delivery 17: 179-96 (1987); "primers as a means to improve the Delivery of peptide drivers", arm v. drug Delivery rev.8 (1): 1-38 (1992); fleisher et al, "Imprvedoral drug delivery: solubility compositions over come by the use of drugs ", arm v. drug Delivery rev.19 (2): 115- & ltSUB & gt 130- & gt (1996); fleisher et al, "Design of drugs for improved Targeting by intracellular Enzyme Targeting", MethodsEnzymol.112(Drug Enzyme Targeting, Pt.A): 360-81, (1985); farquhar D et al, "biologicaily Reversible Phosphate-Protective Groups", pharm.Sci., 72 (3): 324-325 (1983); freeman S et al, "Bioreversible Protection for the phosphorus Group: chemical Stability and Bioactivation of Di (4-acetyl-benzyl) methylphosphonate with Carboxyesterase, "chem.Soc., chem.Commun., 875-; fris and Bundgaard, "precursors of phosphates and phosphates: novel lipophilic alicyclic oxygenated alkyl esters derivatives of phosphate-or phosphate associating drugs masking the negative charges of the group ", eur.j.pharm.sci.4: 49-59 (1996); gangwar et al, "Pro-drug, molecular structure and persistence delivery", Des.Biopharm.Prop.Prodrug Natas, [ Symp. ] recording Date 1976, 409-21. (1977); nathwani and Wood, "Penicillins: a current review of the clinical pharmacologic and therapeutic use ", Drugs 45 (6): 866-94 (1993); sinhababu and Thakker, "produgs of anti agents", adv. drug Delivery rev.19 (2): 241-273 (1996); stella et al, "products, do the y had accommodations in clinical practice? ", Drugs 29 (5): 455-73 (1985); "Development and optimization of anti-HIV nucleosidic analogs and primers: a review of the same cellular pharmacology, structure-activity pharmacologies and pharmacologics ", adv. drug Delivery Rev.39 (1-3): 117-; taylor, "advanced passive real via drivers", adv. drug delivery rev, 19 (2): 131-148 (1996); valentino and Borehardt, "Drug variants of the endogenous responses of peptides", Drug Discovery Today 2 (4): 148-155 (1997); wiebe and Knaus, "topics for the design of anti-HIV nucleosidic HIV infection for treating cephalic HIV infection", adv. drug delivery rev.: 39(1-3): 63-80 (1999); waller et al, "produgs", br.j.clin.pharmac.28: 497-507(1989), all of which are incorporated herein by reference in their entirety.

The term "protected derivative" refers to a derivative of a compound described herein in which the reactive site is blocked by a protecting group. The protected derivatives may be used to prepare the compounds or may be active per se. A comprehensive list of suitable Protecting Groups can be found in T.W.Greene, Protecting Groups in Organic Synthesis, 3 rd edition, John Wiley & Sons, Inc.1999.

"isomers" refers to any compound having the same molecular formula but differing in its nature or in the order of bonding of its atoms or in the arrangement of its atoms in space. Isomers in which the atoms differ in their spatial arrangement are referred to as "stereoisomers". Stereoisomers that are not mirror images of each other are referred to as "diastereomers", and stereoisomers that are mirror images that do not overlap with each other are referred to as "enantiomers" or sometimes "optical isomers". The carbon atoms bonded to the four different substituents are referred to as "chiral centers". Compounds with one chiral center have two enantiomeric forms of opposite chirality. The mixture of the two enantiomeric forms is referred to as a "racemic mixture". The compounds having more than one chiral center have 2^n-1A pair of enantiomers wherein n is the number of chiral centers. Compounds with more than one chiral center can be present as individual diastereomers or as mixtures of enantiomers (known as "diastereomeric mixtures"). When a chiral center is present, stereoisomers can be characterized by the absolute configuration of the chiral center. Absolute configuration refers to the spatial arrangement of substituents attached to a chiral center. Enantiomers are characterized by the absolute configuration of their chiral centers and are described by the R-and S-sequence rules of Cahn, Ingold and Prelog. Stereochemical nomenclature conventions, methods of determining stereochemistry, and methods of separating stereoisomers are well known in the art (see, e.g., "Advanced Organic Chemistry", 4 th edition, March, Jerry, John Wiley&Sons，New York，1992)。

The term "enantiomer" is used to describe one of a pair of molecular isomers that are mirror images of each other and do not overlap. Other terms used to designate or refer to enantiomers include "stereoisomers" (because of different arrangements or stereochemistry around the chiral center; while all enantiomers are stereoisomers, not all stereoisomers are enantiomers) or "optical isomers" (because of the optical activity of pure enantiomers, this is the ability of different pure enantiomers to rotate plane-polarized light in different directions). Enantiomers generally have the same physical properties, such as melting and boiling points, and also have the same spectral characteristics. Enantiomers can differ from each other with respect to their interaction with plane-polarized light and with respect to biological activity.

The labels "R" and "S" are used to indicate the absolute configuration of a molecule with respect to its chiral center. The tag may appear as a prefix or suffix; it may or may not be separated from the isomers by hyphens; which may or may not have hyphens; which may or may not have brackets.

The labels or prefixes "(+)" and "(-) -are used to denote the sign of the compound rotating plane polarized light, where (-) means that the compound is left-handed (rotated to the left). The compound with the prefix (+) is dextrorotatory (rotates to the right).

The terms "racemic mixture", "racemic compound" or "racemate" refer to a mixture of two enantiomers of a compound. An ideal racemic mixture is a 50: 50 mixture of the two enantiomers of the compound such that the optical rotation of the (+) enantiomer cancels out the optical rotation of the (-) enantiomer.

The term "resolving", when used in reference to a racemic mixture, refers to the separation of a racemate into its two enantiomeric forms (i.e., (+) and (-); 65(R) and (S) forms). The term may also refer to the selective conversion of one isomer of the racemate to the product.

The term "enantiomeric excess" or "ee" refers to a reaction product in which one enantiomer is produced in excess of the other, and is a definition of a mixture of the (+) -and (-) -enantiomers, wherein the composition is given in molar or weight or volume fractions F (+) and F (- (wherein the sum of F (+) and F (-) is 1). Enantiomeric excess is defined as F (+) -F (-), and percent enantiomeric excess is defined as 100x F (+) -F (-). The "purity" of an enantiomer is described by the ee or percent ee value (% ee).

Whether expressed as "purified enantiomers" or "pure enantiomers" or "resolved enantiomers" or "enantiomeric excess of a compound," the term is intended to mean that the amount of one enantiomer exceeds the amount of the other. Thus, when referring to an enantiomeric preparation, both (or one) of the percent (e.g., by mole or weight or volume) of the major enantiomer and (or) the percent enantiomeric excess of the major enantiomer can be used to determine whether the preparation represents a pure enantiomeric preparation.

The term "enantiomeric purity" or "enantiomeric purity" of an isomer is a qualitative or quantitative measure of a pure enantiomer; typically, measurements are expressed on the basis of ee or enantiomeric excess.

The terms "substantially pure enantiomers", "substantially resolved enantiomers", "substantially pure enantiomeric preparation" are intended to mean a preparation (e.g., derived from a non-optically active starting material, substrate or intermediate) wherein one enantiomer is enriched relative to the other, and more preferably wherein the other enantiomer constitutes less than 20%, more preferably less than 10%, and more preferably less than 5%, and still more preferably less than 2% of the enantiomeric or enantiomeric preparation.

The terms "pure enantiomer", "resolved enantiomer" and "pure enantiomeric preparation" are intended to mean a preparation (e.g., derived from a non-optically active starting material, substrate or intermediate) in which one enantiomer (e.g., the R enantiomer) is enriched relative to the other, and more preferably in which the other enantiomer (e.g., the S enantiomer) constitutes less than 30%, preferably less than 20%, more preferably less than 10% of the preparation (e.g., in this particular case, the R enantiomer is substantially free of the S enantiomer), and more preferably less than 5%, and still more preferably less than 2%. A pure enantiomer may be synthesized substantially free of the other enantiomer, or a pure enantiomer may be synthesized in a stereopreferred procedure followed by a separation step, or a pure enantiomer may be obtained from a racemic mixture.

The term "enantioselectivity", also known as the enantiomeric ratio indicated by the symbol "E", refers to the ability of an enzyme to selectively produce one enantiomer from a racemic substrate relative to the other in a racemic mixture of the product, in other words, it measures the ability of the enzyme to distinguish between racemates. Nonselective reactions have an E of 1, while resolution with an E greater than 20 is generally considered useful for synthesis or resolution. The enantioselectivity lies in the difference in the conversion rate between the enantiomers in question. The reaction product obtained is enriched in one enantiomer; in contrast, the remaining substrate is enriched in the other enantiomer. For practical purposes, it is often desirable to obtain one of the enantiomers in large quantities. This is achieved by stopping the conversion process when the conversion has progressed to a certain extent.

As used herein, "linker" refers to an organic moiety that links two moieties of a compound. The linker typically comprises a direct bond or atom, e.g. oxygen or sulphur, and a unit, e.g. NR⁴、C(O)、C(O)NH、C(O)O、NHC(O)O、OC(O)O、SO、SO₂、SO₂NH or a chain of atoms, such as substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, arylalkyl, arylalkenyl, arylalkynyl, heteroarylalkyl, heteroarylalkenyl, heteroarylalkynyl, heterocyclylalkyl, heterocyclylalkenyl, heterocyclylalkynyl, aryl, heteroaryl, heterocyclyl, cycloalkyl, cycloalkenyl, alkylarylalkyl, alkylarylalkenyl, alkylarylalkyl, alkenylarylalkyl, alkenylarylalkenyl, alkenylarylalkynyl, alkynylarylalkyl, alkynylarylalkenyl, alkynylarylalkynyl, alkylheteroarylalkyl, alkylheteroarylalkenyl, alkylheteroarylalkynyl, alkenylheteroarylalkyl, alkenylheteroarylalkynyl, alkynylheteroarylalkyl, alkynylheteroarylalkenyl, alkynylalkyl, alkylheterocyclylalkylalkynyl, alkylheteroarylalkynyl, and heteroaryl alkynyl, An alkylheterocyclylalkenyl, an alkylheterocyclylalkynyl, an alkenylheterocyclylalkyl, an alkenylheterocyclenyl, an alkenylheterocyclylalkynyl, an alkynylheterocyclylalkyl, an alkynylheterocyclenyl, an alkynylheterocyclylalkynyl, an alkylaryl, an alkenylaryl, an alkynylaryl, an alkylheteroaryl, an alkenylheteroaryl, an alkynylheteroaryl, wherein one or more methylene groups may be replaced byO、S、S(O)、SO₂、NR⁴、C(O)、C(O)NH、C(O)O、NHC(O)O、OC(O)O、SO₂NH, a cleavable linker, a substituted or unsubstituted aryl, a substituted or unsubstituted heteroaryl, a substituted or unsubstituted heterocyclyl interrupted or terminated; wherein R4 is hydrogen, acyl, aliphatic or substituted aliphatic.

In some embodiments, the linker is a branched linker. The branching point of the branched linker may be at least trivalent, but may also be a tetravalent, pentavalent or hexavalent atom, or a group exhibiting such a multivalence. In some embodiments, the branch point is-N, -N (Q) -C, -O-C, -S-C, -SS-C, -C (O) N (Q) -C, -OC (O) N (Q) -C, -N (Q) C (O) -C, or-N (Q) C (O) O-C; wherein each occurrence of Q is independently H or optionally substituted alkyl. In some embodiments, the branch point is glycerol or a derivative thereof.

The cleavable linker is sufficiently stable extracellularly, but is cleaved after entry into the target cell to release the two moieties that the linker holds together. In a preferred embodiment, the cleavable linker is cleaved at least 10-fold or more, preferably at least 100-fold or more faster in the target cells or under first reference conditions (which may, for example, be selected to mimic or represent intracellular conditions) than in the blood or serum of the subject or under second reference conditions (which may, for example, be selected to mimic or represent conditions present in the blood or serum).

The cleavable linker is sensitive to a cleaving agent (e.g., pH, redox potential, or presence of a degrading molecule). Typically, the lytic agent is more prevalent or present at a higher level or activity within the cell than in serum or blood. Examples of such degradation agents include: redox agents selected for a particular substrate or not having substrate specificity, including, for example, oxidizing or reducing enzymes or reducing agents present in the cell, such as thiols, which can degrade the redox cleavable linker by reduction; an esterase; an amidase; endosomes or agents that can create an acidic environment, e.g., those that result in a pH of 5 or less; peptidases (which may be substrate specific) and proteases, as well as phosphatases, by acting as enzymes that generally hydrolyze or degrade acid cleavable linkers.

The linker may comprise a cleavable linker that is cleaved by a particular enzyme. The enzymatically cleavable linker may be an enzyme substrate that is cleaved by an enzyme. Such substrates are also referred to herein as cleavable enzyme substrates. The type of cleavable linker incorporated into the linker may depend on the cell to be targeted. When the target cell type is peptidase-rich, a linker comprising a peptide bond may be used.

In some embodiments, the linker may include a cleavable linker that is cleavable by cathepsin G. Figure 11 shows an exemplary molecule cleaved by cathepsin G.

In some embodiments, the cleavable linker is cleaved at least 1.25, 1.5, 1.75, 2, 3, 4, 5, 10, 25, 50, or 100 times faster intracellularly (or under in vitro conditions selected to mimic intracellular conditions) as compared to blood or serum (or under in vitro conditions selected to mimic extracellular conditions). In some embodiments, the cleavable linker cleaves less than 90%, 80%, 70%, 60%, 50%, 40%, 30%, 20%, 10%, 5%, or 1% in blood (or under in vitro conditions selected to mimic extracellular conditions) as compared to in cells (or under in vitro conditions selected to mimic intracellular conditions).

Exemplary cleavable linkers include, but are not limited to, redox cleavable linkers (e.g., -S-S-and-C (R))₂-S-S-, wherein R is H or C₁-C₆Alkyl, and at least one R is C₁-C₆Alkyl radicals, e.g. CH₃Or CH₂CH₃) (ii) a Phosphate-based cleavable linkers (e.g., -O-P (O) (OR) -O-, -O-P (S) (SR) -O-, -S-P (O) (OR) -O-, -O-P (O) (OR) -S-, -S-P (O) -S-, -O-P (S) (ORk) -S-, -S-P (S) (OR) -O-, -O-P (O) (R) -O-, -O-P (O) -O-, -O-P (S) (R) -O-, -S-P (O) (R) -O-, (R-), (R) -S-P (S), (R) -O-, -S-P (O) (R) -S-, -O-P (S), (R) -S-, (O-P) (O) (OH) -O-, -O-P (S) (SH) -O-, -S-P (O) (OH) -O-, -O-P (O) (OH) -S-, -S-P (OH) -O-, -O-P (OH) -S-, -S-P (O) (OH) -H-S-, -O-P (S) (OH) -S-, -S-P (S) (OH) -O-, -O-P (O) (H) -O-, -O-P (S) (H) -O-, -S-P (O) (H) -O-, -S-P (S) (H) -O-, -S-P (O) (H) -S-and-O-P (S) (H) -S-, wherein R is optionally substituted straight chain or branched chain C₁-C₁₀Alkyl groups); acidic cleavable linkers (e.g., hydrazones, esters and esters of amino acids, -C ═ NN-and-oc (o)); an ester-based cleavable linker (e.g., -c (O) O-); peptide-based cleavable linkers, (e.g., linkers that are cleaved by intracellular enzymes such as peptidases and proteases, e.g., -NHCHR^AC(O)NHCHR^BC (O) -, wherein R^AAnd R^BIs the R group of two adjacent amino acids). The peptide-based cleavable linker comprises two or more amino acids. In some embodiments, the peptide-based cleavable linker comprises an amino acid sequence that is a substrate for a peptidase or protease present in the cell.

In some embodiments, the acidic cleavable linker is cleavable under acidic conditions at a pH of about 6.5 or less (e.g., about 6.5, 6.0, 5.5, 5.0 or less), or by an agent such as an enzyme that can be a generic acid.

Linkers according to the present invention also include prodrug moieties and nanoparticles. For non-limiting example, the prodrug moiety may be a linker susceptible to "cleavage" to produce the active form of the drug. More information can be found in: bundgard (1985, Design of produgs, pages 7-9, pages 21-24, Elsevier, Amsterdam) and Silverman (1992, the Organic Chemistry of Drug Design and Drug Action, pages 352 and 401, Academic Press, San Diego, Calif.), which are incorporated herein by reference in their entirety as if fully set forth.

In some embodiments, the cleavable linker is cleavable by an enzyme present in greater amounts in cancer cells or tumors than in non-cancer cells or normal cells. For example, the cleavable linker is cleavable by a peptidase or protease present in greater amounts in pancreatic cancer cells. In one embodiment, the linker comprises a compound shown in figure 11.

Unless otherwise defined herein, scientific and technical terms used in connection with the present application shall have the commonly understood meaning as understood by one of ordinary skill in the art to which this disclosure belongs. It is to be understood, however, that this invention is not limited to the particular methodology, protocols, reagents, etc. described herein as these may vary. The terminology used herein is for the purpose of describing particular embodiments only and is not intended to limit the scope of the present invention, which is defined only by the claims.

One problem addressed by the present invention is related to the spread of cancer (e.g., pancreatic cancer) through metastasis and the emergence of resistance of cancer to treatment. One disadvantage of current agents used to slow cancer cell proliferation is that these agents transform cells into cancer stem-like cells, which are more likely to develop resistance to targeted therapies, and are more likely to metastasize and spread tumors. Currently, there is no solution to prevent cancer metastasis. The present invention provides compounds, compositions, methods and kits that inhibit cancer progression through metastasis and reduce the appearance of dry-like features.

Dual inhibitor compounds

In various embodiments, the present invention provides compounds that inhibit both HDAC and GSK3 β. Compounds that inhibit both HDAC and GSK3 β are also referred to herein as dual inhibitors.

In embodiments of the various aspects disclosed herein, the dual inhibitor compound is of formula (IV):

wherein:

L₁and L₂Independently a linker;

R¹is an aromatic moiety, alkyl, acyl, cyclyl or heterocyclyl, each of which may be optionally substituted;

R²is hydrogen, alkyl, cyclic, heterocyclicAryl or heteroaryl, each of which may be optionally substituted;

R³is absent or is an aromatic moiety, which may be optionally substituted;

p is 0, 1, 2, 3, 4, 5, 6,7, 8, 9, 10, 11 or 12; and is

One of which. R¹-L₁-one nitrogen attached to the thiadiazolidine ring, and- (CH)₂)_p-R³-L₂-C(O)NHOR²Another nitrogen attached to the thiadiazolidine ring.

In some other embodiments of the aspects disclosed herein, the compound is of formula (V):

wherein:

L₁and L₂Independently a linker;

R¹is an aromatic moiety, alkyl, acyl, cyclyl or heterocyclyl, each of which may be optionally substituted;

R²is hydrogen, lower alkyl, cyclyl, heterocyclyl, aryl or heteroaryl, each of which may be optionally substituted;

R³is absent or is an aromatic moiety, which may be optionally substituted; and is

p is 0, 1, 2, 3, 4, 5, 6,7, 8, 9, 10, 11 or 12.

In each compound of formula (IV) or (V), R¹May be an optionally substituted aryl or lower alkyl group. In some embodiments, R in formula (IV) or (V)¹Can be independently selected from C₁-C₁₀Alkyl, aryl or heteroaryl, each of which may be optionally substituted with 1, 2, 3 or 4 substituents.

In various embodiments, R¹May be a lower alkyl group. In some embodiments, R₁May be C₁-C₆An alkyl group. R¹Exemplary alkyl groups of (a) include, but are not limited to, methyl, ethyl, propyl, isopropyl, butyl, sec-butyl, isobutyl, tert-butyl, pentyl, neopentyl, and hexyl. In one embodiment, R¹Is methyl.

In various embodiments, R¹Is optionally substituted aryl or optionally substituted heteroaryl selected from: pyridyl, pyrimidinyl, furyl, thienyl, imidazolyl, thiazolyl, pyrazolyl, pyridazinyl, pyrazinyl, triazinyl, tetrazolyl, indolyl, benzyl, phenyl, naphthyl, anthracyl, azulenyl, fluorenyl, indanyl, indenyl, naphthyl, phenyl, tetrahydronaphthyl, benzimidazolyl, benzofuranyl, benzothiofuranyl, benzothiophenyl, benzoxazolyl, benzoxazolinyl, benzothiazolyl, benzotriazolyl, benzotetrazolyl, benzisoxazolyl, benzisothiazole, benzimidazolinyl, carbazolyl, 4aH carbazolyl, carbolinyl, chromanyl, chromenyl, cinnolinyl, decahydroquinolinyl, 2H, 6H-1, 5, 2-diazinyl, dihydrofuro [2, 3b ] 2]Tetrahydrofuran, furyl, furazanyl, imidazolidinyl, imidazolinyl, imidazolyl, 1H-indazolyl, indolinyl, indolizinyl, indolyl, 3H-indolyl, isatoiyl, isobenzofuran, isochroman, isoindolyl, isoquinolinyl, isothiazolyl, isoxazolyl, methylenedioxyphenyl, morpholinyl, naphthyridinyl, octahydroisoquinolinyl, oxadiazolyl, 1, 2, 3-oxadiazolyl, 1, 2, 4-oxadiazolyl, 1, 2, 5-oxadiazolyl, 1, 3, 4-oxadiazolyl, oxazolidinyl, oxazolyl, oxindolyl, pyrimidinyl, phenanthridinyl, phenanthrolinyl, phenazinyl, phenothiazinyl, phenoxazinyl, phthalazinyl, piperazinyl, piperidinyl, piperidyl, piperidonyl, 4-piperidonyl, pyridonyl, Piperonyl, pteridinyl, purinyl, pyranyl, pyrazinyl, pyrazolidinyl, pyrazolinyl, pyrazolyl, pyridazinyl, pyridooxazolyl, pyridoimidazolyl, pyridylThiazolyl, pyridyl, pyrimidyl, pyrrolidinyl, pyrrolinyl, 2H-pyrrolyl, quinazolinyl, quinolinyl, 4H-quinolizinyl, quinoxalinyl, quinuclidinyl, tetrahydrofuryl, tetrahydroisoquinoline, tetrahydroquinolinyl, tetrazolyl, 6H-1, 2, 5-thiadiazinyl, 1, 2, 3-thiadiazolyl, 1, 2, 4-thiadiazolyl, 1, 2, 5-thiadiazolyl, 1, 3, 4-thiadiazolyl, thianthrenyl, thiazolyl, thienyl, thiazolothiazolyl, thiazolooxazolyl, thiazoloimidazolyl, thienyl, and xanthenyl.

In some embodiments, R1 is optionally substituted phenyl. Typically, the optionally substituted phenyl group may be substituted with 1, 2, 3, 4 or 5 substituents independently selected from: alkyl, CF₃、NO₂、CO₂H、SO₂H. Cyano, hydroxy, mercapto, alkylthio, alkoxy, acyl, halogen, amino, alkylamino, dialkylamino, and any combination thereof. Preferably, the optionally substituted phenyl is substituted with one substituent. In one embodiment, the optionally substituted phenyl is 4-methoxyphenyl.

In various embodiments of formula (IV) or (V), R²May be hydrogen, lower alkyl, 3-8 membered cyclic or heterocyclic group or 5-8 membered aryl or heteroaryl, each of which may be optionally substituted. In some embodiments, R²Is hydrogen or lower alkyl. In various embodiments R²May be C₁-C₆An alkyl group. R²Exemplary alkyl groups of (a) include, but are not limited to, methyl, ethyl, propyl, isopropyl, butyl, sec-butyl, isobutyl, tert-butyl, pentyl, neopentyl, and hexyl. In some embodiments, R²Is H or methyl.

In various embodiments of formula (IV) or (V), R³May be absent or an optionally substituted aryl or optionally substituted heteroaryl. R₃Exemplary optionally substituted aryl and optionally substituted heteroaryl groups of (a) include, but are not limited to: pyridyl, pyrimidinyl, furyl, thienyl, imidazolyl, thiazolyl, pyrazolyl, pyridazinyl, pyrazinyl, triazinyl, tetrazolylA phenyl group, an indenyl group, a benzyl group, a phenyl group, a naphthyl group, an anthracenyl group, an azulenyl group, a fluorenyl group, an indanyl group, an indenyl group, a naphthyl group, a phenyl group, a tetrahydronaphthyl group, a benzimidazolyl group, a benzofuranyl group, a benzothiofuranyl group, a benzothiophenyl group, a benzoxazolyl group, a benzoxazolinyl group, a benzothiazolyl group, a benzotriazolyl group, a benzotetrazolyl group, a benzisoxazolyl group, a benzisothiazole, a benzimidazolinyl group, a carbazolyl group, a 4aH carbazolyl group, a carbolinyl group, a chromanyl group, a chromenyl group, a cinnolinyl group, a decahydroquinolyl group, a 2H, 6H-1, 5, 2-diazinyl group]Tetrahydrofuran, furyl, furazanyl, imidazolidinyl, imidazolinyl, imidazolyl, 1H-indazolyl, indolinyl, indolizinyl, indolyl, 3H-indolyl, isatoiyl, isobenzofuran, isochroman, isoindolyl, isoquinolinyl, isothiazolyl, isoxazolyl, methylenedioxyphenyl, morpholinyl, naphthyridinyl, octahydroisoquinolinyl, oxadiazolyl, 1, 2, 3-oxadiazolyl, 1, 2, 4-oxadiazolyl, 1, 2, 5-oxadiazolyl, 1, 3, 4-oxadiazolyl, oxazolidinyl, oxazolyl, oxindolyl, pyrimidinyl, phenanthridinyl, phenanthrolinyl, phenazinyl, phenothiazinyl, phenoxazinyl, phthalazinyl, piperazinyl, piperidinyl, piperidyl, piperidonyl, 4-piperidonyl, pyridonyl, Piperonyl, pteridinyl, purinyl, pyranyl, pyrazinyl, pyrazolidinyl, pyrazolinyl, pyrazolyl, pyridazinyl, pyridooxazolyl, pyridoimidazolyl, pyridothiazolyl, pyridyl, pyrimidinyl, pyrrolidinyl, pyrrolinyl, 2H-pyrrolyl, quinazolinyl, quinolinyl, 4H-quinolizinyl, quinoxalinyl, quinuclidinyl, tetrahydrofuranyl, tetrahydroisoquinolinyl, tetrazolyl, 6H-1, 2, 5-thiadiazinyl, 1, 2, 3-thiadiazolyl, 1, 2, 4-thiadiazolyl, 1, 2, 5-thiadiazolyl, 1, 3, 4-thiadiazolyl, thianthrenyl, thiazolyl, thienyl, thiazolothiazolyl, thiazolooxazolyl, thiazoloimidazolyl, thienyl, and xanthenyl.

In some embodiments, R3 is optionally substituted phenyl. The optionally substituted phenyl group may be substituted with 1, 2, 3 or 4 substituents independently selected from: alkyl, CF₃、NO₂、CO₂H、SO₂H. Cyano, hydroxy, mercapto, alkylthio, alkoxy, acyl, halogen, amino, alkylamino, dialkylamino, and any combination thereof. Preferably, the optionally substituted phenyl is substituted with one substituent.

In some further embodiments, R³Is absent.

In various embodiments of formula (IV) or (V), p is 0, 1, 2, 3, or 5, preferably, p is 0 or 1. In some embodiments, p is 0. In some further embodiments, p is 1.

In various embodiments, the linker is independently selected from the group consisting of a bond, - (CH) at each occurrence₂)_q-、-(CH₂)_qCH＝CH(CH₂)_r-、-NH-、-NHC(O)(CH₂)_q-and any combination thereof, wherein q is independently at each occurrence 1, 2, 3, 4, 5, 6,7, 8, 9, 10, 11, or 12, and wherein r is independently at each occurrence 0, 1, 2, 3, 4, 5, 6,7, 8, 9, 10, 11, or 12.

In various embodiments, L₁Selected from the group consisting of a bond, - (CH)₂)_q-、-(CH₂)_qCH＝CH(CH₂)_r-、-NH-、-NHC(O)(CH₂)_q-and any combination thereof, wherein q is independently at each occurrence 1, 2, 3, 4, 5, 6,7, 8, 9, 10, 11, or 12, and wherein r is independently at each occurrence 0, 1, 2, 3, 4, 5, 6,7, 8, 9, 10, 11, or 12. In some embodiments, L is₁Is a bond or- (CH)₂)_q-。L₁Preferred q values of (a) include, but are not limited to, 1, 2, 3, 4, 5 and 6. In some embodiments, L is₁is-CH₂-. In some further embodiments, L₁Is a bond. In yet other embodiments, L₁is-NH-

In various embodiments, L₂May be selected from the group consisting of a bond, - (CH)₂)_q-、-(CH₂)_qCH＝CH(CH₂)_r-、-NH-、-NHC(O)(CH₂)_q-and any combination thereof, wherein q is independently at each occurrence 1, 2, 3, 4, 5, 6,7, 8, 9, 10, 11, or 12, and wherein r is independently at each occurrence 0, 1, 2, 3, 4, 5, 6,7, 8, 9, 10, 11, or 12. In some embodiments, L is₂Is a bond; -NH-; -NHC (O) (CH)₂)_q-, wherein q is 4, 6 or 8; or-CH₂CH ═ CH-. In some embodiments, L is₂Is a bond. In some further embodiments, L₂is-NH-.

In the various compounds disclosed herein, L₁-R₁is-CH₂-phenyl. In some other embodiments, L₁-R₁Is CH₃. In still other embodiments, L₁-R₁Is 4-methoxybenzyl.

In some embodiments, p is 0 or 1, and R₃Is phenyl. In some further embodiments, p is 0 and R³Is absent.

In some compounds, p is 0; r³Is phenyl; and L is₂Is a bond or-NHC (O) (CH)₂)_q-group, wherein q is 4, 6 or 8. In some other embodiments, p is 1; r³Is phenyl; l is₂is-NHC (O) (CH)₂)_q-, wherein q is 6. In still other embodiments, p is 0 and L₂is-CH₂CH＝CH-。

In some embodiments, the compound of formula (IV) is a compound of formula (VI):

wherein R is¹、R²、R³、L₁、L₂And p is as defined for formula (IV).

In some embodiments, the compound of formula (IV) is a compound of formula (VII):

wherein R is¹、R²、R³、L₁、L₂And p is as defined for formula (IV).

In various embodiments, the compound according to formula (IV) is a compound of formula (I):

wherein X is a linker (e.g., L)₂) And Y is absent or a substituent of an aromatic group. In some embodiments, Y is selected from alkyl, CF₃、NO₂、CO₂H、SO₂H. Cyano, hydroxy, mercapto, alkylthio, alkoxy, acyl, halogen, amino, alkylamino, dialkylamino, and any combination thereof. Although only one Y substituent is shown, more than one Y may be present on the phenyl ring, for example, one, two, three, four or five Y. In some embodiments, Y is absent.

In various embodiments, X may be selected from the group consisting of a bond, - (CH)₂)_q-、-(CH₂)_qCH＝CH(CH₂)_r-、-NH-、-NHC(O)(CH₂)_q-and any combination thereof, wherein q is independently at each occurrence 1, 2, 3, 4, 5, 6,7, 8, 9, 10, 11, or 12, and wherein r is independently at each occurrence 0, 1, 2, 3, 4, 5, 6,7, 8, 9, 10, 11, or 12. In some embodiments, X is a bond; -NH-; -NHC (O) (CH)₂)_q-, wherein q is 4, 6 or 8; or-CH₂CH ═ CH-. In some embodiments, X is-NH-.

In some embodiments, the compound of formula (I) is a compound of formula (I-1):

wherein n is 1, 2, 3, 4, 5, 6,7, 8, 9, 10, 11 or 12.

In various embodiments of formula (I-1), n is 4, 6 or 8, i.e., a compound of formula (I-1a), (I-1b) or (I-1 c):

the compound of formula (I-1a) is also referred to herein as ALB-185602. The compound of formula (I-1b) is also referred to herein as ALB-185644. The compound of formula (I-1c) is also referred to herein as ALB-185643.

In some embodiments, the compound of formula (I) is a compound of formula (I-2):

in some embodiments, the compound of formula (I) is a compound of formula (I-3):

in various embodiments, the compound of formula (IV) is a compound of formula (II):

wherein X is a linker group and R is-L₁R¹。

In a number of embodiments, the first and second electrodes are,x may be selected from the group consisting of a bond, - (CH)₂)_q-、-(CH₂)_qCH＝CH(CH₂)_r-、-NH-、-NHC(O)(CH₂)_q-and any combination thereof, wherein q is independently at each occurrence 1, 2, 3, 4, 5, 6,7, 8, 9, 10, 11, or 12, and wherein r is independently at each occurrence 0, 1, 2, 3, 4, 5, 6,7, 8, 9, 10, 11, or 12. In some embodiments, X is a bond; -NH-; -NHC (O) (CH)₂)_q-, wherein q is 4, 6 or 8; or-CH₂CH ═ CH-. In some embodiments, X is-NH-.

In various embodiments, -L₁R¹May be selected from the bond-R¹、-(CH₂)_q-R¹、-(CH₂)_qCH＝CH(CH₂)_r-R¹、-NH-R¹、-NHC(O)(CH₂)_q-R¹And any combination thereof, wherein q is independently at each occurrence 1, 2, 3, 4, 5, 6,7, 8, 9, 10, 11, or 12, and wherein r is independently at each occurrence 0, 1, 2, 3, 4, 5, 6,7, 8, 9, 10, 11, or 12. In some embodiments, -L₁R¹Is a bond-R¹；-NH-R¹；-NHC(O)(CH₂)_q-R¹Wherein q is 4, 6 or 8; or-CH₂CH＝CH-R¹。

In some embodiments of compounds of formula (II), -L₁R¹Is an optionally substituted alkyl group. In one embodiment, R is methyl.

Exemplary compounds of formula (II) include, but are not limited to, compounds of formula (II-1):

in various embodiments, the compound of formula (V) is a compound of formula (III):

wherein X is a linker (e.g., L)₂) And Y is absent or a substituent of an aromatic group. In some embodiments, Y is selected from alkyl, CF₃、NO₂、CO₂H、SO₂H. Cyano, hydroxy, mercapto, alkylthio, alkoxy, acyl, halogen, amino, alkylamino, dialkylamino, and any combination thereof. Although only one Y substituent is shown, more than one Y may be present on the phenyl ring, for example, one, two, three, four or five Y.

In various embodiments, at least one Y is present and is alkoxy. Exemplary alkoxy groups for Y include, but are not limited to, methoxy, ethoxy, propoxy, tert-butoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, and the like. In one embodiment, Y is methoxy.

In various embodiments, X may be selected from the group consisting of a bond, - (CH)₂)_q-、-(CH₂)_qCH＝CH(CH₂)_r-、-NH-、-NHC(O)(CH₂)_q-and any combination thereof, wherein q is independently at each occurrence 1, 2, 3, 4, 5, 6,7, 8, 9, 10, 11, or 12, and wherein r is independently at each occurrence 0, 1, 2, 3, 4, 5, 6,7, 8, 9, 10, 11, or 12. In some embodiments, X is a bond; -NH-; -NHC (O) (CH)₂)_q-, wherein q is 4, 6 or 8; or-CH₂CH ═ CH-. In some embodiments, X is a bond.

In some embodiments, the compound of formula (III) is a compound of formula (III-1):

the compound of formula (III-1) is also referred to herein as ALB-185357. In various embodiments, the compound of formula (V) is a compound of formula (IIIb):

wherein X is a linker (e.g., L)₂) And Y is absent or a substituent of an aromatic group. In some embodiments, Y is selected from alkyl, CF₃、NO₂、CO₂H、SO₂H. Cyano, hydroxy, mercapto, alkylthio, alkoxy, acyl, halogen, amino, alkylamino, dialkylamino, and any combination thereof. Although only one Y substituent is shown, more than one Y may be present on the phenyl ring, for example, one, two, three, four or five Y.

In various embodiments, at least one Y is present and is alkoxy. Exemplary alkoxy groups for Y include, but are not limited to, methoxy, ethoxy, propoxy, tert-butoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, and the like. In one embodiment, Y is methoxy.

In various embodiments, X may be selected from the group consisting of a bond, - (CH)₂)_q-、-(CH₂)_qCH＝CH(CH₂)_r-、-NH-、-NHC(O)(CH₂)_q-and any combination thereof, wherein q is independently at each occurrence 1, 2, 3, 4, 5, 6,7, 8, 9, 10, 11, or 12, and wherein r is independently at each occurrence 0, 1, 2, 3, 4, 5, 6,7, 8, 9, 10, 11, or 12. In some embodiments, X is a bond; -NH-; -NHC (O) (CH)₂)_q-, wherein q is 4, 6 or 8; or-CH₂CH ═ CH-. In some embodiments, X is a bond.

In some embodiments, the compound of formula (IIIb) is a compound of formula (IIIb-1) having the structure:

the compound of formula (IIIb-1) is also referred to herein as ALB-188540.

The compounds disclosed herein can be prepared starting from commercially available starting materials using general synthetic techniques and procedures known to those skilled in the art. Chemicals are commercially available from companies such as Aldrich, Argonaut Technologies, VWR and Lancaster. Chromatography consumables and equipment are commercially available from companies such as: AnaLogix, Inc, Burlington, wis; biotage AB, Charlottesville, Va.; analytical Sales and Services, inc., pomptonpilins, n.j.; teledyne Isco, Lincoln, nebr; VWR International, Bridgeport, n.j.; varian inc, Palo Alto, calif, and Multigram II Mettler Toledo Instrument Newark, del.

The synthesis of various exemplary compounds of formula (IV) is shown in schemes I, II, III, and IV below, as well as the synthesis of some exemplary compounds of formula (V) is shown in scheme V below. It should be noted that any of these may be readily employed by one skilled in the art to prepare compounds of formulae (I) - (VI). Scheme I describes a reaction scheme towards compounds of formula I, such as targets 1a, 1b and 1c (formulas I-1a, 1b and 1c), in various embodiments according to the present invention. Scheme II depicts a reaction scheme towards a compound of formula I, such as target 2 (formula I-2), according to various embodiments of the present invention. Target 2 was similar to the En Vivo Pharma HDAC inhibitor shown in figure 8. Scheme III describes a reaction scheme towards a compound of formula I, such as target 3 (formula I-3), according to various embodiments of the present invention. Target 3 has a smaller zinc binding moiety. Scheme IV depicts a reaction scheme towards a compound of formula II, e.g., target 4 (formula II-1), according to various embodiments of the present invention. Target 4 combines the benzamide moiety of SAHA and the benzyl moiety of TDZD-8. Scheme V depicts a reaction scheme towards a compound of formula III, such as target 5 (formula III-1), according to various embodiments of the present invention. Object 5 is an analog of the Khanfar et al reported GSK3 beta inhibitor (Discovery of novel GSK-3 beta inhibitors with a potential in vitro and in vivo activity and expression library synthesized ligand-and structure-based viral screening; J Med chem.2010Dec 23; 53 (24): 8534-45), which is incorporated herein by reference in its entirety as if fully set forth herein. The hydroxamic acid moiety of target 5 may participate in the key H bond required for GSK3 β activity and act as a zinc binding moiety for HDAC inhibition.

Targets 1a, 1b and 1c

Scheme I

Object 2

Scheme II

Target 3

Scheme III

Target 4

Scheme IV

Target 5

Scheme V

As discussed herein, the dual inhibitor, HDAC inhibitor, or GSK3 β inhibitor may be conjugated to a particle, e.g., a magnetic particle. In some embodiments, the dual inhibitor, HDAC inhibitor, or GSK3 β inhibitor may be attached to the particle through a linker comprising a cleavable group. For example, the linker may comprise a group that can cleave at a higher rate in a cancer cell or tumor relative to its cleavage in a non-cancer cell. In some embodiments, the linker may comprise a group cleavable by an enzyme present in higher amounts in cancer cells or tumors relative to the amount in non-cancer cells. In some embodiments, the linker may comprise a cleavable group that is cleaved by a peptidase that is present in higher amounts in cancer cells or tumors relative to the amount in non-cancer cells. In some embodiments, the linker comprises a cleavable group that is cleaved by cathepsin G.

Fig. 10-12 show components of an exemplary system having a dual inhibitor (fig. 10) or each of an HDAC inhibitor and a GSK3 β inhibitor, and a prodrug moiety such as a cleavable enzyme substrate and a nanoparticle. For example, a dual inhibitor or each of an HDAC inhibitor and a GSK3 β inhibitor is attached to a peptide substrate (fig. 11), which in turn can be attached to magnetic particles (fig. 12). Attachment to the magnetic carrier allows for the introduction of the agent to the tumor. Since tumors have a high concentration of cathepsin G, the inhibitor is released at the tumor.

As non-limiting examples, inhibitors (e.g., dual inhibitors, or each of an HDAC inhibitor and a GSK3 β inhibitor) can be attached to a cleavable enzyme substrate (e.g., cathepsin G substrate, Suc-AAPF-pNA from Santa Cruz Biotechnology) using protocols recommended by the manufacturer and/or known to one of ordinary skill in the art. As non-limiting examples, a cleavable enzyme substrate (e.g., cathepsin G substrate Suc-AAPF-pNA from Santa Cruz Biotechnology) can be attached to a magnetic particle (e.g., siMAG from Chemicell) using protocols recommended by the manufacturer and/or known to one of ordinary skill in the art (e.g., free carboxyl conjugation, the carbodiimide method, and mannich reaction).

In general, the particles may be of any shape or form, for example, spherical, rod-like, ellipsoidal, cylindrical, capsule, or disc; and these particles may be part of a network or aggregate. Without limitation, the particles may have any size from nm to millimeters. In some embodiments, the particle is a microparticle or a nanoparticle. As used herein, the term "microparticle" refers to a particle having a particle size of about 1 μm to about 1000 μm. As used herein, the term "nanoparticle" refers to a particle having a particle size of about 0.1nm to about 1000 nm. Generally, the particles disclosed herein are nanoparticles and have an average diameter of about 5nm to about 500 nm. In some embodiments, the average diameter of the particles is from about 75nm to about 500nm, from about 25nm to about 250nm, from about 50nm to about 150nm, from about 75nm to about 125nm, from about 50nm to about 500nm, from about 75nm to about 200nm, from about 100 to about 175nm, from about 125nm to about 175nm, from about 40nm to about 90nm, or from about 50nm to about 80 nm.

In some embodiments, the nanoparticles can be less than about 1um in diameter, for example, about 1um in diameter or less, about 500nm in diameter or less, about 400nm in diameter or less, about 300nm in diameter or less, about 200nm in diameter or less, about 100nm in diameter or less, about 50nm in diameter or less, or about 10nm in diameter or less. In some embodiments, the nanoparticles can be less than 1um in diameter, for example, 1um in diameter or less, 500nm in diameter or less, 400nm in diameter or less, 300nm in diameter or less, 200nm in diameter or less, 100nm in diameter or less, 50nm in diameter or less, or 10nm in diameter or less. In some embodiments, the nanoparticles in the composition can be from about 1nm to about 1um in diameter, for example from about 1nm to about 500nm in diameter, from about 1nm to about 200nm in diameter, from about 10nm to about 200nm in diameter, from about 100nm to about 200nm in diameter, or from about 10nm to about 100nm in diameter. In some embodiments, the nanoparticles in the composition can be 1nm to 1um in diameter, for example 1nm to 500nm in diameter, 1nm to 200nm in diameter, 10nm to 200nm in diameter, 100nm to 200nm in diameter, or 10nm to 100nm in diameter.

In some embodiments, nanoparticles of a particular size may be selected, for example, less than about 200nm in diameter. Methods of selecting nanoparticles of a particular size and/or size range are known in the art and may include, by way of non-limiting example, filtration, sedimentation, centrifugation, and/or chromatography, e.g., SEC.

It will be appreciated by those of ordinary skill in the art that the particles generally exhibit a particle size distribution of about the "size" indicated. The term "particle size" as used herein, unless otherwise indicated, refers to the manner in which the particles are distributed, i.e., the values that occur most often in the particle size distribution. Methods for measuring particle size are known to the skilled person, for example by dynamic light scattering (such as light correlation spectroscopy), laser diffraction, Low Angle Laser Light Scattering (LALLS) and Medium Angle Laser Light Scattering (MALLS)), light obscuration methods (e.g. Coulter analysis) or other techniques (such as rheology, and light or electron microscopy).

In some embodiments, the particles may be substantially spherical. By "substantially spherical" is meant that the ratio of the longest to shortest vertical axis of the cross-section of the particle is less than or equal to about 1.5. A substantially spherical shape does not require a line of symmetry. Furthermore, the particles may have a surface texture, such as lines or depressions or protrusions of small size compared to the overall size of the particle, and still be substantially spherical. In some embodiments, the length ratio between the longest axis and the shortest axis of the particle is less than or equal to about 1.5, less than or equal to about 1.45, less than or equal to about 1.4, less than or equal to about 1.35, less than or equal to about 1.30, less than or equal to about 1.25, less than or equal to about 1.20, less than or equal to about 1.15, less than or equal to about 1.1. Without wishing to be bound by theory, minimizing surface contact in substantially spherical particles minimizes undesirable agglomeration of particles upon storage. Many crystals or platelets have flat surfaces which allow for greater surface contact area where agglomeration can occur through ionic or non-ionic interactions. The spherical shape allows for a smaller area of contact.

The particles may be, for example, monodisperse or polydisperse, and the diameter of a given dispersed particle may vary. In some embodiments, the particles have substantially the same particle size. Particles with a broad size distribution have both relatively large particles and relatively small particles, which allow the smaller particles to fill the gaps between the larger particles, creating new contact surfaces. A broad size distribution can produce larger spheres by creating many contact opportunities for binding agglomeration. The particles described herein are within a narrow size distribution, thereby minimizing the chance of contact agglomeration. "narrow particle size distribution" means a particle size distribution in which the ratio of the volume diameter of the 90 th percentile of the small spherical particles to the volume diameter of the 10 th percentile is less than or equal to 5. In some embodiments, the ratio of the volume diameter of the small spherical particles in the 90 th percentile to the volume diameter in the 10 th percentile is less than or equal to 4.5, less than or equal to 4, less than or equal to 3.5, less than or equal to 3, less than or equal to 2.5, less than or equal to 2, less than or equal to 1.5, less than or equal to 1.45, less than or equal to 1.40, less than or equal to 1.35, less than or equal to 1.3, less than or equal to 1.25, less than or equal to 1.20, less than or equal to 1.15, or less than or equal to 1.1.

Geometric Standard Deviation (GSD) can also be used to indicate narrow particle size distribution. GSD calculations involved determining the Effective Cutoff Diameter (ECD) at accumulations of less than 15.9% and 84.1%. GSD is equal to the square root of the ratio of ECD less than 84.17% to ECD less than 15.9%. When GSD < 2.5, GSD has a narrow particle size distribution. In some embodiments, the GSD is less than 2, less than 1.75, or less than 1.5. In one embodiment, the GSD is less than 1.8.

In various embodiments, the particles may comprise a magnetic material. As used herein, the term "magnetic material" refers to a material or substance that is affected by a magnetic field, i.e., the relative permeability (μ) of the material_r) Greater than 1. Such magnetic materials are intended to include those referred to as ferromagnetic, diamagnetic, paramagnetic, and superparamagnetic. As a general understanding of the terms, superparamagnetic materials exhibit magnetic properties only when subjected to an externally applied magnetic field, otherwise exhibit substantially no magnetic properties; and its total magnetic properties are greater than the sum of the individual particles considered individually. If the particle size of the magnetic material is small enough, the magnetic material will likely be superparamagnetic.The magnetic properties of particles containing magnetic material are greatly influenced by the saturation magnetization, size and concentration of the magnetic material as well as the strength of the external magnetic field.

The magnetic material may be any molecule, composition, particle, or substance that exhibits magnetic properties when incorporated into a matrix. The magnetic material may be selected from the group of elements having atomic numbers 21-29, 42, 44 and 57-70, with elements having atomic numbers 24-29 or 62-69 being particularly preferred. Preferably, the magnetic material is selected from the group consisting of, but not limited to, rare earth metals (such as gadolinium, terbium, dysprosium, holmium, erbium, and europium), transition metals (such as iron, nickel, cobalt, magnesium chromium, and copper), noble metals (such as rhodium, palladium), oxides, compositions, combinations, solid dispersions, and alloys thereof.

In some embodiments, the magnetic material is selected from maghemite (Fe)₂O₃) Magnetite (Fe)₃O₄) Strontium ferrite, samarium-cobalt, neodymium-iron-boron (NIB), magnetite, pyrrhotite, BaFe₁₂O₁₉Alnico magnet alloys, decamethyl metallocenes, and transfer salts of 7, 7, 8, 8-Tetracyanoquinodimethane (TCNQ) or Tetracyanoethylene (TCNE) (e.g. [ Fe (Cp) ])₂]⁺[TCNE]^-、[Fe(Cp*)₂]⁺[TCNQ]^-、[Cr(Cp*)₂]⁺[TCNE]^-、[Cr(Cp*)₂]⁺[TCNQ]^-、[Mn(Cp*)₂]⁺[TCNE]^-And [ Mn (Cp) ]₂]⁺[TCNQ]^-) Hexylammonium trichloroketonate (II) (CuCl)₃(C₆H₁₁NH₃) An Fe-based amorphous magnetic powder, and combinations thereof.

In some embodiments, the particles comprising a magnetic material are magnetic nanoparticles. Magnetic nanoparticles are a class of nanoparticles that can be manipulated using a magnetic field. Such particles are usually composed of magnetic elements such as iron, nickel and cobalt and chemical compounds thereof. Magnetic nanoparticles are well known and methods for their synthesis are described in the art, for example, in U.S. patent nos. 6,878,445; no.5,543,158; no.5,578,325; no.6,676,729; nos. 6,045,925 and 7,462,446, and U.S. patent publication No. 2005/0025971; no. 2005/0200438; no. 2005/0201941; no. 2005/0271745; no. 2006/0228551; no. 2006/0233712; no.2007/01666232 and No.2007/0264199, the contents of all of which are incorporated herein by reference in their entirety.

In some embodiments, the particles are siMAG magnetic beads, available, for example, from chemical GmbH (Berlin, Germany). Two amine-functionalized and carboxy-functionalized SiMAG beads were obtained. Thus, any method using amine or carboxyl coupling can be used to conjugate dual inhibitors, HDAC inhibitors or GSK3 β inhibitors to the siMAG beads. For example, carbodiimide-based coupling reactions can be used for amine-functionalized and carboxy-functionalized siMAG beads. Mannich reactions can be used for amine functionalized siMAG beads.

Method of treatment

In various embodiments, the present invention provides methods of treating a disorder, preventing a disorder, reducing the likelihood of developing a disorder, reducing the severity of a disorder, and/or slowing the progression of a disorder in a subject. The method consists of, or consists essentially of, or comprises: administering to the subject a therapeutically effective amount of a dual inhibitor, thereby treating, preventing, reducing the likelihood of, reducing the severity of, and/or slowing the progression of the disorder in the subject.

In various embodiments, the methods further comprise administering or treating with one or more additional anti-cancer therapies. Examples of anti-cancer therapies include, but are not limited to, surgery, radiation therapy (radiotherapy), biological therapy, immunotherapy, chemotherapy, or combinations of these therapies. In addition, cytotoxic, anti-angiogenic and antiproliferative agents may be used in combination with dual inhibitors.

In those embodiments where a combination therapy regimen is used, the dual inhibitor and one or more anti-cancer therapeutic agents described herein can be administered in a therapeutically effective amount or a therapeutically synergistic amount. As used in such embodiments, including combination therapies, a therapeutically effective amount is such that co-administration of the dual inhibitor and one or more other anti-cancer therapeutic agents results in reduction or inhibition of cancer as described herein. A "therapeutically synergistic amount" is an amount of dual inhibitor and one or more other anti-cancer therapeutic agents required to synergistically or significantly reduce or eliminate a disorder or symptom associated with a particular cancer.

In some embodiments, the dual inhibitor and the one or more additional anti-cancer therapeutic agents may be administered simultaneously or sequentially at a time and in amounts sufficient to reduce or eliminate the occurrence or recurrence of a tumor, dormant tumor, or micrometastases. In some embodiments, the dual inhibitor and one or more other therapeutic agents may be administered as maintenance therapy to prevent tumor recurrence or reduce the likelihood of tumor recurrence.

Without limitation, the dual inhibitor and the one or more additional anti-cancer therapeutic agents may be provided separately in separate compositions or in the same composition. In addition, the dual inhibitor and the one or more other anti-cancer therapeutic agents may be administered simultaneously or sequentially. In certain embodiments, the dual inhibitor is administered before, during, or after administration of one or more other anti-cancer therapeutic agents.

In various embodiments, the method further comprises: administering a chemotherapeutic agent to the subject. In some embodiments, the dual inhibitor and chemotherapeutic agent are provided in one composition. In other embodiments, the dual inhibitor and the chemotherapeutic agent are provided in different compositions. In various embodiments, the dual inhibitor and the chemotherapeutic agent are administered simultaneously or sequentially. In certain embodiments, the dual inhibitor is administered before, during, or after administration of the chemotherapeutic agent.

As understood by one of ordinary skill in the art, suitable doses of chemotherapeutic agents or other anti-cancer agents are generally about those already used in clinical therapy, e.g., where the chemotherapeutic agent is administered alone or in combination with other chemotherapeutic agents. Depending on the condition being treated, variations in dosage may occur. The physician administering the treatment will be able to determine the appropriate dosage for the individual subject.

In addition to the treatment regimens described above, the subject may be treated with radiation.

In other embodiments, the dual inhibitor is attached to a cleavable enzyme substrate, and the cleavable enzyme substrate is attached to the magnetic particle. In one embodiment, the cleavable enzyme substrate is a substrate for an enzyme that is enriched in the cancer or tumor. In another embodiment, the cleavable enzyme substrate is a substrate for a peptidase enzyme that is enriched in cancer or tumors. In certain embodiments, the cleavable enzyme substrate is a substrate for cathepsin G. In various embodiments, the method further comprises directing the dual inhibitor to the cancer or tumor using a magnetic field.

In some embodiments, the dual inhibitor is a compound of formula IV, formula V, formula VI, or formula VII. In various embodiments, the dual inhibitor is a compound of formula I, formula II, formula III, IIIb, formula I-1a, formula I-1b, formula I-1c, formula I-2, formula I-3, formula II-1, formula III-1, formula IIIb-1, or a combination thereof.

According to the present invention, the dual inhibitor may be administered using a suitable mode of administration, for example, the manufacturer's recommended mode of administration. In accordance with the present invention, dual inhibitors of the claimed method may be administered using a variety of routes including, but not limited to, aerosol, nasal, oral, transmucosal, transdermal, parenteral, implantable pump, continuous infusion, topical administration, capsule, and/or injection. In various embodiments, the dual inhibitor is administered topically, intravascularly, intravenously, intraarterially, intratumorally, intramuscularly, subcutaneously, intraperitoneally, intranasally, or orally.

Typical dosages of effective amounts of dual inhibitors when using known therapeutic compounds are within the manufacturer's recommended ranges, and are also indicated to the skilled artisan as in vitro responses in cells or in vivo responses in animal models. Such dosages can generally be reduced by up to about one order of magnitude in terms of concentration and amount, without loss of relevant biological activity. The actual dosage may depend on the judgment of the practitioner, the condition of the patient and the effectiveness of the treatment method based on, for example, the in vitro responsiveness of the relevant cultured cells or tissue cultured tissue sample, or the response observed in a suitable animal model. In various embodiments, the dual inhibitor may be administered once per day (SID/QD), twice per day (BID), three times per day (TID), four times per day (QID), or more, such that an effective amount of the dual inhibitor is administered to the subject, wherein the effective amount is any one or more of the dosages described herein.

In some embodiments, the dual inhibitor is administered as follows: about 0.001-0.01, 0.01-0.1, 0.1-0.5, 0.5-5, 5-10, 10-20, 20-50, 50-100, 100-200, 200-300, 300-400, 400-500, 500-600, 600-700, 700-800, 800-900 or 900-1000mg/kg or a combination thereof. In other embodiments, the dual inhibitor is administered as follows: about 0.001-0.01, 0.01-0.1, 0.1-0.5, 0.5-5, 5-10, 10-20, 20-50, 50-100, 100-200, 200-300, 300-400, 400-500, 500-600, 600-700, 700-800, 800-900 or 900-1000 μ g/kg or a combination thereof. In various embodiments, the dual inhibitor is administered about 1-3 times per day, 1-7 times per week, or 1-9 times per month. In various embodiments, the dual inhibitor is administered for about 1-10 days, 10-20 days, 20-30 days, 30-40 days, 40-50 days, 50-60 days, 60-70 days, 70-80 days, 80-90 days, 90-100 days, 1-6 months, 6-12 months, or 1-5 years. In various embodiments, the dual inhibitor is administered once, twice, three times, or more.

In various embodiments, an effective amount of a dual inhibitor is any one or more of: about 0.001-0.01, 0.01-0.1, 0.1-0.5, 0.5-5, 5-10, 10-20, 20-50, 50-100, 100-200, 200-300, 300-400, 400-500, 500-600, 600-700, 700-800, 800-900 or 900-1000 μ M or a combination thereof.

In various embodiments, an effective amount of a dual inhibitor is any one or more of: about 0.01 to 0.05. mu.g/kg/day, 0.05 to 0.1. mu.g/kg/day, 0.1 to 0.5. mu.g/kg/day, 0.5 to 5. mu.g/kg/day, 5 to 10. mu.g/kg/day, 10 to 20. mu.g/kg/day, 20 to 50. mu.g/kg/day, 50 to 100. mu.g/kg/day, 100 to 150. mu.g/kg/day, 150 to 200. mu.g/kg/day, 200 to 250. mu.g/kg/day, 250 to 300. mu.g/kg/day, 300 to 350. mu.g/kg/day, 350 to 400. mu.g/kg/day, 400 to 500. mu.g/kg/day, 500 to 600. mu.g/kg/day, 600 to 700. mu.g/kg/day, 700 to 800. mu.g/kg/day, 800 to 900. mu.g/kg/day, 900 to 1000. mu.g/kg/day, 0.01 to 0.05 mg/kg/day, 0.05-0.1 mg/kg/day, 0.1 to 0.5 mg/kg/day, 0.5 to 1 mg/kg/day, 1 to 5 mg/kg/day, 5 to 10 mg/kg/day, 10 to 15 mg/kg/day, 15 to 20 mg/kg/day, 20 to 50 mg/kg/day, 50 to 100 mg/kg/day, 100 to 200 mg/kg/day, 200 to 300 mg/kg/day, 300 to 400 mg/kg/day, 400 to 500 mg/kg/day, 500 to 600 mg/kg/day, 600 to 700 mg/kg/day, 700 to 800 mg/kg/day, 800 to 900 mg/kg/day, 900 to 1000 mg/kg/day, or combinations thereof. Herein, "μ g/kg/day" or "mg/kg/day" means μ g or mg per kg body weight of the subject per day.

In various embodiments, the present invention provides methods of treating a disorder, preventing a disorder, reducing the likelihood of developing a disorder, reducing the severity of a disorder, and/or slowing the progression of a disorder in a subject. The method consists of or consists essentially of or comprises: administering to the subject a therapeutically effective amount of an HDAC inhibitor and a GSK3 β inhibitor, thereby treating, preventing, reducing the likelihood of, reducing the severity of, and/or slowing the progression of the disorder in the subject.

In some embodiments, the HDAC inhibitor and GSK3 β inhibitor are provided in one composition. In other embodiments, the HDAC inhibitor and GSK3 β inhibitor are provided in different compositions. In various embodiments, the HDAC inhibitor and GSK3 β inhibitor are administered simultaneously or sequentially. In certain embodiments, the HDAC inhibitor is administered before, during, or after the GSK3 β inhibitor.

In various embodiments, in addition to administering the HDAC inhibitor and the GSK3 β inhibitor, the methods further comprise administering or treating one or more additional cancer therapies. In those embodiments where a combination therapy regimen is used, the HDAC inhibitor, GSK3 β inhibitor and one or more anti-cancer therapeutic agents described herein are administered in therapeutically effective amounts or therapeutically synergistic amounts. As used in such embodiments, including combination therapies, therapeutically effective amounts are such that co-administration of the HDAC inhibitor, GSK3 β inhibitor and one or more other anti-cancer therapeutic agents results in the reduction or inhibition of cancer as described herein. In this context, a "therapeutically synergistic amount" is the amount of HDAC inhibitor and GSK3 β inhibitor and one or more other anti-cancer therapeutic agents required to synergistically or significantly reduce or eliminate the condition or symptom associated with a particular cancer.

In some embodiments, the HDAC inhibitor, GSK3 β inhibitor, and one or more other anti-cancer therapeutic agents may be administered simultaneously or sequentially at a time and in amounts sufficient to reduce or eliminate the occurrence or recurrence of a tumor, dormant tumor, or micrometastases. In some embodiments, the HDAC inhibitor, GSK3 β inhibitor, and one or more other therapeutic agents may be administered as a maintenance therapy to prevent tumor recurrence or reduce the likelihood of tumor recurrence.

Without limitation, at least one of the HDAC inhibitor or GSK3 β inhibitor and the one or more other anti-cancer therapeutic agents may be provided separately in separate compositions or in the same composition. In addition, the dual inhibitor and the one or more other anti-cancer therapeutic agents may be administered simultaneously or sequentially. In certain embodiments, the dual inhibitor is administered before, during, or after administration of one or more other anti-cancer therapeutic agents.

In some embodiments, at least one of the HDAC inhibitor and/or GSK3 β inhibitor and the additional anti-cancer therapeutic agent are provided in the same composition. In some embodiments, the HDAC inhibitor, GSK3 β inhibitor, and the additional anti-cancer therapeutic are provided in a composition. In other embodiments, the HDAC inhibitor, GSK3 β inhibitor, and the additional anti-cancer therapeutic agent are provided in different compositions. In various embodiments, the HDAC inhibitor, GSK3 β inhibitor, and the additional anti-cancer therapeutic are administered simultaneously or sequentially. In certain embodiments, at least one of the HDAC inhibitor or GSK3 β inhibitor is administered before, during, or after the administration of the additional anti-cancer therapeutic.

In various embodiments, the method further comprises: administering to the subject a chemotherapeutic agent in addition to the HDAC inhibitor and the GSK3 β inhibitor. In some embodiments, at least one of the HDAC inhibitor and/or GSK3 β inhibitor and the additional chemotherapeutic agent are provided in the same composition. In some embodiments, the HDAC inhibitor, GSK3 β inhibitor, and the additional chemotherapeutic agent are provided in a composition. In other embodiments, the HDAC inhibitor, GSK3 β inhibitor, and chemotherapeutic agent are provided in different compositions. In various embodiments, the HDAC inhibitor, GSK3 β inhibitor, and chemotherapeutic agent are administered simultaneously or sequentially. In certain embodiments, at least one of the HDAC inhibitor and GSK3 β inhibitor is administered before, during, or after the chemotherapeutic agent.

In addition to administering the HDAC inhibitor and the GSK3 β inhibitor, the subject may be treated with radiation.

In further embodiments, the HDAC inhibitor and/or GSK3 β inhibitor is attached to a cleavable enzyme substrate, and the cleavable enzyme substrate is attached to a magnetic particle. In one embodiment, the cleavable enzyme substrate is a substrate for an enzyme that is enriched in the cancer or tumor. In another embodiment, the cleavable enzyme substrate is a substrate for a peptidase enzyme that is enriched in cancer or tumors. In certain embodiments, the cleavable enzyme substrate is a substrate for cathepsin G. In various embodiments, the method further comprises directing the HDAC inhibitor and/or GSK3 β inhibitor to the cancer or tumor using a magnetic field.

In various embodiments, the HDAC inhibitor is SAHA, TSA, TPX, MS-275, valproic acid, or CHAP31, or a functional equivalent, analog, derivative, or salt thereof, or a combination thereof. In various embodiments, the GSK3 β inhibitor is SB216763, TDZD-8, or Tideglusib (NP-12), or a functional equivalent, analog, derivative, or salt thereof, or a combination thereof.

According to the present invention, the HDAC inhibitor and GSK3 β inhibitor may be administered using an appropriate mode of administration, for example, the mode of administration recommended by the manufacturer for each of the HDAC inhibitor and GSK3 β inhibitor. According to the present invention, the HDAC inhibitor and GSK3 β inhibitor of the claimed methods may be administered using a variety of routes, including but not limited to aerosol, nasal, oral, transmucosal, transdermal, parenteral, implantable pump, continuous infusion, topical administration, capsule and/or injection. In various embodiments, the HDAC inhibitor is administered topically, intravascularly, intravenously, intraarterially, intratumorally, intramuscularly, subcutaneously, intraperitoneally, intranasally, or orally. In various embodiments, the GSK3 β inhibitor is administered topically, intravascularly, intravenously, intraarterially, intratumorally, intramuscularly, subcutaneously, intraperitoneally, intranasally, or orally.

Typical dosages of effective amounts of HDAC inhibitors and/or GSK3 β inhibitors when using known therapeutic compounds are within the manufacturer's recommended ranges and are also indicated to the skilled person as in vitro responses in cells or in vivo responses in animal models. Such dosages can generally be reduced by up to about one order of magnitude in terms of concentration and amount, without loss of relevant biological activity. The actual dosage may depend on the judgment of the practitioner, the condition of the patient and the effectiveness of the treatment method based on, for example, the in vitro responsiveness of the relevant cultured cells or tissue cultured tissue sample, or the response observed in a suitable animal model. In various embodiments, the HDAC inhibitor and/or GSK3 β inhibitor may be administered once per day (SID/QD), twice per day (BID), three times per day (TID), four times per day (QID) or more, such that an effective amount of the HDAC inhibitor and/or GSK3 β inhibitor is administered to the subject, wherein the effective amount is any one or more of the dosages described herein.

In various embodiments, the HDAC inhibitor is administered as follows: about 0.001-0.01, 0.01-0.1, 0.1-0.5, 0.5-5, 5-10, 10-20, 20-50, 50-100, 100-200, 200-300, 300-400, 400-500, 500-600, 600-700, 700-800, 800-900 or 900-1000mg/kg or a combination thereof. In other embodiments, the HDAC inhibition is administered as follows: about 0.001-0.01, 0.01-0.1, 0.1-0.5, 0.5-5, 5-10, 10-20, 20-50, 50-100, 100-200, 200-300, 300-400, 400-500, 500-600, 600-700, 700-800, 800-900 or 900-1000 μ g/kg or a combination thereof. In various embodiments, the HDAC inhibitor is administered from about 1-3 times per day, 1-7 times per week, or 1-9 times per month. In various embodiments, the HDAC inhibitor is administered for about 1-10 days, 10-20 days, 20-30 days, 30-40 days, 40-50 days, 50-60 days, 60-70 days, 70-80 days, 80-90 days, 90-100 days, 1-6 months, 6-12 months, or 1-5 years. In various embodiments, the HDAC inhibitor is administered once, twice, three times, or more. In one embodiment, the HDAC inhibitor is SAHA, or a functional equivalent, analog, derivative or salt of SAHA.

In various embodiments, the GSK3 β inhibitor is administered as follows: about 0.001-0.01, 0.01-0.1, 0.1-0.5, 0.5-5, 5-10, 10-20, 20-50, 50-100, 100-200, 200-300, 300-400, 400-500, 500-600, 600-700, 700-800, 800-900 or 900-1000mg/kg or a combination thereof. In other embodiments, GSK3 β inhibition is administered as follows: about 0.001-0.01, 0.01-0.1, 0.1-0.5, 0.5-5, 5-10, 10-20, 20-50, 50-100, 100-200, 200-300, 300-400, 400-500, 500-600, 600-700, 700-800, 800-900 or 900-1000 μ g/kg or a combination thereof. In various embodiments, the GSK3 β inhibitor is administered about 1-3 times per day, 1-7 times per week, or 1-9 times per month. In various embodiments, the GSK3 β inhibitor is administered for about 1-10 days, 10-20 days, 20-30 days, 30-40 days, 40-50 days, 50-60 days, 60-70 days, 70-80 days, 80-90 days, 90-100 days, 1-6 months, 6-12 months, or 1-5 years. In various embodiments, the GSK3 β inhibitor is administered one, two, three, or more times. In one embodiment, the GSK3 β inhibitor is TDZD-8, or a functional equivalent, analog, derivative, or salt of TDZD-8. In one embodiment, the GSK3 β inhibitor is Tideglusib, or a functional equivalent, analog, derivative, or salt thereof.

In various embodiments, the effective amount of the HDAC inhibitor and/or GSK3 β inhibitor is any one or more of: about 0.001-0.01, 0.01-0.1, 0.1-0.5, 0.5-5, 5-10, 10-20, 20-50, 50-100, 100-200, 200-300, 300-400, 400-500, 500-600, 600-700, 700-800, 800-900 or 900-1000 μ M or a combination thereof.

In various embodiments, the effective amount of the HDAC inhibitor and/or GSK3 β inhibitor is any one or more of the following: about 0.01 to 0.05. mu.g/kg/day, 0.05 to 0.1. mu.g/kg/day, 0.1 to 0.5. mu.g/kg/day, 0.5 to 5. mu.g/kg/day, 5 to 10. mu.g/kg/day, 10 to 20. mu.g/kg/day, 20 to 50. mu.g/kg/day, 50 to 100. mu.g/kg/day, 100 to 150. mu.g/kg/day, 150 to 200. mu.g/kg/day, 200 to 250. mu.g/kg/day, 250 to 300. mu.g/kg/day, 300 to 350. mu.g/kg/day, 350 to 400. mu.g/kg/day, 400 to 500. mu.g/kg/day, 500 to 600. mu.g/kg/day, 600 to 700. mu.g/kg/day, 700 to 800. mu.g/kg/day, 800 to 900. mu.g/kg/day, 900 to 1000. mu.g/kg/day, 0.01 to 0.05 mg/kg/day, 0.05-0.1 mg/kg/day, 0.1 to 0.5 mg/kg/day, 0.5 to 1 mg/kg/day, 1 to 5 mg/kg/day, 5 to 10 mg/kg/day, 10 to 15 mg/kg/day, 15 to 20 mg/kg/day, 20 to 50 mg/kg/day, 50 to 100 mg/kg/day, 100 to 200 mg/kg/day, 200 to 300 mg/kg/day, 300 to 400 mg/kg/day, 400 to 500 mg/kg/day, 500 to 600 mg/kg/day, 600 to 700 mg/kg/day, 700 to 800 mg/kg/day, 800 to 900 mg/kg/day, 900 to 1000 mg/kg/day, or combinations thereof. As used herein, "μ g/kg/day" or "mg/kg/day" means μ g or mg per kilogram body weight of a subject per day

In various embodiments, the subject is a human. In some embodiments, the subject is a mammalian subject, including but not limited to human, monkey, ape, dog, cat, cow, horse, goat, pig, rabbit, mouse, and rat.

In various embodiments, the disorder is a cancer or tumor. In some embodiments, the disease is pancreatic cancer. In some embodiments, the dual inhibitor, HDAC inhibitor and/or GSK3 β inhibitor may be administered during the prophylactic phase of the disorder (i.e., when the subject has not yet developed the disorder, but is likely to develop the disorder or is in the process of developing the disorder). In other embodiments, the dual inhibitor, HDAC inhibitor and/or GSK3 β inhibitor may be administered during the therapeutic phase of the disorder (i.e., when the subject already develops the disorder). As a non-limiting example, the target disorder is pancreatic cancer. In this exemplary case, the patient may be treated with the methods described herein if the patient has not yet developed, or is likely to develop, or is in the process of developing, or already has developed pancreatic cancer.

Pharmaceutical composition

In various embodiments, the present invention provides compositions consisting of, consisting essentially of, or comprising dual inhibitors of HDAC and GSK3 β. According to the present invention, the composition may be used to treat a disorder, prevent a disorder, reduce the likelihood of developing a disorder, reduce the severity of a disorder, and/or slow the progression of a disorder in a subject.

In various embodiments, the dual inhibitor is a compound of formula I, formula II, formula III, formula I-1a, formula I-1b, formula I-1c, formula I-2, formula I-3, formula II-1, formula III-1, formula IIIb-1, formula IV, formula V, formula VI, formula VII, or a combination thereof. In some embodiments, the dual inhibitor in the composition is provided at the following mg of dual inhibitor per kilogram body weight of the subject: for example, about 0.001-0.01, 0.01-0.1, 0.1-0.5, 0.5-5, 5-10, 10-20, 20-50, 50-100, 100-200, 200-300, 300-400, 400-500, 500-600, 600-700, 700-800, 800-900 or 900-1000mg/kg or the combination thereof. In other embodiments, the dual inhibitors in the composition are provided as the following μ g of dual inhibitor per kilogram of subject body weight: about 0.001-0.01, 0.01-0.1, 0.1-0.5, 0.5-5, 5-10, 10-20, 20-50, 50-100, 100-200, 200-300, 300-400, 400-500, 500-600, 600-700, 700-800, 800-900 or 900-1000 μ g/kg or a combination thereof.

In various further embodiments, the composition further comprises a cleavable enzyme substrate and a magnetic particle, wherein the dual inhibitor is attached to the cleavable enzyme substrate and the cleavable enzyme substrate is attached to the magnetic particle. In one embodiment, the cleavable enzyme substrate is a substrate for an enzyme that is enriched in the cancer or tumor. In another embodiment, the cleavable enzyme substrate is a substrate for a peptidase enzyme that is enriched in cancer or tumors. In certain embodiments, the cleavable enzyme substrate is a substrate for cathepsin G.

In various embodiments, the present invention provides compositions consisting of, consisting essentially of, or comprising an HDAC inhibitor and a GSK3 β inhibitor. According to the present invention, the composition may be used to treat a disorder, prevent a disorder, reduce the likelihood of developing a disorder, reduce the severity of a disorder, and/or slow the progression of a disorder in a subject.

In various embodiments, the HDAC inhibitor is SAHA, TSA, TPX, MS-275, valproic acid, or CHAP31, or a functional equivalent, analog, derivative, or salt thereof, or a combination thereof. In some embodiments, the HDAC inhibitor in the composition is provided at the following mg of HDAC inhibitor per kilogram body weight of the subject: for example, about 0.001-0.01, 0.01-0.1, 0.1-0.5, 0.5-5, 5-10, 10-20, 20-50, 50-100, 100-200, 200-300, 300-400, 400-500, 500-600, 600-700, 700-800, 800-900 or 900-1000mg/kg or the combination thereof. In other embodiments, the HDAC inhibitor in the composition is provided at the following μ g of HDAC inhibitor per kilogram body weight of the subject: about 0.001-0.01, 0.01-0.1, 0.1-0.5, 0.5-5, 5-10, 10-20, 20-50, 50-100, 100-200, 200-300, 300-400, 400-500, 500-600, 600-700, 700-800, 800-900 or 900-1000 μ g/kg or a combination thereof.

In various embodiments, the GSK3 β inhibitor is SB216763, TDZD-8, or Tideglusib (NP-12), or a functional equivalent, analog, derivative, or salt thereof, or a combination thereof. In some embodiments, the GSK3 β inhibitor in the composition is provided as the following mg of GSK3 β inhibitor per kilogram body weight of the subject: for example, about 0.001-0.01, 0.01-0.1, 0.1-0.5, 0.5-5, 5-10, 10-20, 20-50, 50-100, 100-200, 200-300, 300-400, 400-500, 500-600, 600-700, 700-800, 800-900 or 900-1000mg/kg or the combination thereof. In other embodiments, the GSK3 β inhibitor in the composition is provided as the following μ g of GSK3 β inhibitor per kilogram subject body weight: for example, about 0.001-0.01, 0.01-0.1, 0.1-0.5, 0.5-5, 5-10, 10-20, 20-50, 50-100, 100-200, 200-300, 300-400, 400-500, 500-600, 600-700, 700-800, 800-900 or 900-1000 μ g/kg or combinations thereof.

In various further embodiments, the composition further comprises a cleavable enzyme substrate and a magnetic particle, wherein the HDAC inhibitor and/or GSK3 β inhibitor is attached to the cleavable enzyme substrate, and the cleavable enzyme substrate is attached to the magnetic particle. In one embodiment, the cleavable enzyme substrate is a substrate for an enzyme that is enriched in the cancer or tumor. In another embodiment, the cleavable enzyme substrate is a substrate for a peptidase enzyme that is enriched in cancer or tumors. In certain embodiments, the cleavable enzyme substrate is a substrate for cathepsin G.

In certain embodiments, a plurality of the compositions described herein further comprise a chemotherapeutic agent. In some embodiments, the chemotherapeutic agent is selected from the group consisting of actinomycin, alitretinol, all-trans retinoic acid, azacitidine, azathioprine, bevacizumab, bexarotene, bleomycin, bortezomib, carboplatin, capecitabine, cetuximab, cisplatin, chlorambucil, cyclophosphamide, cytarabine, daunorubicin, docetaxel, doxifluridine, doxorubicin, epirubicin, epothilone, erlotinib, etoposide, fluorouracil, gefitinib, gemcitabine, hydroxyurea, idarubicin, imatinib, yipima, irinotecan, mechlorethamine, melphalan, mercaptopurine, methotrexate, mitoxantrone, Ocrelizumab (Ocrelizumab), ofatumumab, oxaliplatin, paclitaxel, panitumumab, pemetrexed, rituximab, tafeoquin, teniposide, thioguanine, topotecan, Tretinoin, valrubicin, vemurafenib, vinblastine, vincristine, vindesine, vinorelbine, vorinostat, romidepsin, 5-fluorouracil (5-FU), 6-mercaptopurine (6-MP), cladribine, clofarabine, floxuridine, fludarabine, pentostatin, mitomycin, ixabepilone, estramustine, prednisone, methylprednisolone, dexamethasone, or combinations thereof.

In certain embodiments, the pharmaceutical composition according to the invention is administered to a mammal or human. Preferred pharmaceutical compositions also exhibit minimal toxicity when administered to a mammal or human. In various embodiments, the pharmaceutical composition according to the present invention is formulated for topical, intravascular, intravenous, intraarterial, intratumoral, intramuscular, subcutaneous, intraperitoneal, intranasal, or oral administration.

In various embodiments, the pharmaceutical compositions according to the present invention may be formulated for delivery by any route of administration. The "route of administration" may refer to any route of administration known in the art, including, but not limited to, aerosol, nasal, oral, transmucosal, transdermal, parenteral, enteral, topical, or topical. "parenteral" refers to a route of administration typically associated with injection, including intraorbital, infusion, intra-arterial, intracapsular, intracardiac, intradermal, intramuscular, intraperitoneal, intrapulmonary, intraspinal, intrasternal, intrathecal, intrauterine, intravenous, subarachnoid, subcapsular, subcutaneous, transmucosal, or transtracheal. By parenteral route, the compositions may be in the form of solutions or suspensions for infusion or injection, or as lyophilized powders. By the enteral route, the pharmaceutical composition may be in the form of tablets, gel capsules, sugar-coated tablets, syrups, suspensions, solutions, powders, granules, emulsions, microspheres or nanospheres or lipid or polymer vesicles that allow controlled release. By topical route, the pharmaceutical composition may be in the form of an aerosol, lotion, cream, gel, ointment, suspension, solution or emulsion. Methods of administration are known to those skilled in the art.

The pharmaceutical composition according to the invention may be delivered in a therapeutically effective amount. The precise therapeutically effective amount is that amount of the composition which produces the most effective result in terms of therapeutic efficacy in a given subject. Such amounts depend on a variety of factors including, but not limited to, the nature of the therapeutic compound (including activity, pharmacokinetics, pharmacodynamics, and bioavailability), the physiological condition of the subject (including age, sex, type and stage of disease, general physical condition, responsiveness to a given dose, and type of drug), the nature of the pharmaceutically acceptable carrier in the formulation, and the route of administration. One skilled in the clinical and pharmacological arts will be able to determine a therapeutically effective amount by routine experimentation, for example, by monitoring the subject's response to administration of the compound and adjusting the dosage accordingly. For additional guidance, see Remington: the Science and Practice of Pharmacy (edited by Gennaro, 20 th edition, Williams & Wilkins PA, USA) (2000).

In various embodiments, the composition is administered 1-3 times per day, 1-7 times per week, or 1-9 times per month. In various embodiments, the composition is administered for about 1-10 days, 10-20 days, 20-30 days, 30-40 days, 40-50 days, 50-60 days, 60-70 days, 70-80 days, 80-90 days, 90-100 days, 1-6 months, 6-12 months, or 1-5 years. In various embodiments, the dual inhibitor is administered once, twice, three times, or more. In various embodiments, the composition may be administered once daily (SID/QD), twice daily (BID), three times daily (TID), four times daily (QID), or more, so as to administer to the subject an effective amount of the dual inhibitor, the HDAC inhibitor, and/or the GSK3 β inhibitor, wherein the effective amount is any one or more of the dosages described herein.

In various embodiments, the pharmaceutical composition according to the present invention may comprise any pharmaceutically acceptable excipient. "pharmaceutically acceptable excipient" refers to an excipient that can be used to prepare generally safe, non-toxic, and desirable pharmaceutical compositions, and includes excipients that are acceptable for veterinary use as well as for human pharmaceutical use. Such excipients may be solid, liquid, semi-solid, or in the case of aerosol compositions, gaseous. Examples of excipients include, but are not limited to: starches, sugars, microcrystalline cellulose, diluents, granulating agents, lubricants, binders, disintegrating agents, wetting agents, emulsifying agents, coloring agents, releasing agents, coating agents, sweetening agents, flavoring agents, preservatives, antioxidants, plasticizers, gelling agents, thickening agents, hardening agents, styling agents, suspending agents, surfactants, insulating agents, carriers, stabilizing agents, and combinations thereof.

In various embodiments, the pharmaceutical composition according to the present invention may comprise any pharmaceutically acceptable carrier. As used herein, a "pharmaceutically acceptable carrier" refers to a pharmaceutically acceptable material, composition or vehicle involved in carrying or transporting a compound of interest from one tissue, organ or body part to another. For example, the carrier may be a liquid or solid filler, diluent, excipient, solvent or encapsulating material, or a combination thereof. Each component of the carrier must be "pharmaceutically acceptable" in that it must be compatible with the other ingredients of the formulation. It must also be suitable for use in contact with any tissue or organ with which it may come into contact, which means that it cannot carry the risk of toxicity, irritation, allergic response, immunogenicity or any other complications that outweigh its therapeutic benefit.

The pharmaceutical composition according to the invention may also be formulated as capsules, tablets or as an emulsion or syrup for oral administration. Pharmaceutically acceptable solid or liquid carriers may be added to enhance or stabilize the composition, or to aid in the preparation of the composition. Liquid carriers include syrup, peanut oil, olive oil, glycerin, saline, alcohol and water. Solid carriers include starch, lactose, calcium sulfate, dihydrate, terra alba, magnesium stearate or stearic acid, talc, pectin, acacia, agar or gelatin. The carrier may also include a slow release material such as glyceryl monostearate or glyceryl distearate alone or with a wax.

The pharmaceutical formulations may be prepared according to conventional pharmaceutical techniques, including dry milling, mixing and blending for powder forms; for grinding, mixing, granulating and, if necessary, compressing in tablet form; or for milling, mixing and filling in the form of hard gelatin capsules. When a liquid carrier is used, the preparation will be in the form of a syrup, elixir, emulsion or aqueous or non-aqueous suspension. Such liquid formulations may be administered directly orally or filled into soft gelatin capsules.

An adjuvant (formulant) may be added to the composition prior to administration to a patient. Liquid formulations may be preferred. For example, these adjuvants may include oils, polymers, vitamins, carbohydrates, amino acids, salts, buffers, albumin, surfactants, bulking agents, or combinations thereof.

Carbohydrate adjuvants include sugars or sugar alcohols, such as mono-, di-or polysaccharides, or water-soluble glucans. The saccharide or glucan may include fructose, dextrose, lactose, glucose, mannose, sorbose, xylose, maltose, sucrose, dextran, pullulan, dextrin, alpha and beta cyclodextrins, soluble starch, hydroxyethyl starch and carboxymethyl cellulose, or mixtures thereof. "sugar alcohols" are defined as C4 to C8 hydrocarbons having-OH groups and include galactitol, inositol, mannitol, xylitol, sorbitol, glycerol, and arabitol. These sugars or sugar alcohols mentioned above may be used alone or in combination. The amount used is not fixedly limited as long as the sugar or sugar alcohol is soluble in the aqueous preparation. In one embodiment, the sugar or sugar alcohol concentration is between 1.0 w/v% and 7.0 w/v%, more preferably between 2.0 and 6.0 w/v%.

Amino acid adjuvants include carnitine, arginine and betaine in the levorotatory (L) form; however, other amino acids may be added.

The polymeric adjuvant comprises polyvinylpyrrolidone (PVP) having an average molecular weight between 2000 and 3000, or polyethylene glycol (PEG) having an average molecular weight between 3000 and 5000.

It is also preferred to use a buffer in the composition to minimize the change in pH of the solution prior to lyophilization or after reconstitution. Most any physiological buffer can be used, including but not limited to citrate, phosphate, succinate, and glutamate buffers or mixtures thereof. In some embodiments, the concentration is 0.01 to 0.3 molar. Surfactants that can be added to the formulations are shown in EP nos. 270,799 and 268,110.

Another drug delivery system for extending the circulatory half-life is liposomes. Methods for preparing liposome delivery systems are described in Gabizon et al, Cancer Research (1982) 42: 4734; cafioso, Biochem biophysis Acta (1981) 649: 129; and Szoka, Ann Rev Biophys Eng (1980) 9: 467. Other DRUG delivery systems are known in the art and are described, for example, in Poznansky et al, DRUG DELIVERY SYSTEMS (R.L.Juliano, ed., Oxford, N.Y.1980), pp.253-315; m.l. poznansky, Pharm Revs (1984) 36: 277, respectively.

After the liquid pharmaceutical composition is prepared, it may be lyophilized to prevent degradation and to maintain sterility. Methods of lyophilizing liquid compositions are known to those of ordinary skill in the art. Prior to extemporaneous use, the compositions may be reconstituted with a sterile diluent (e.g. ringer's solution, distilled water or sterile saline), which may contain additional ingredients. After rehydration, the composition is administered to the subject using methods known to those skilled in the art.

The composition of the present invention can be sterilized by a conventional well-known sterilization technique. The resulting solution may be packaged for use or filtered under sterile conditions and lyophilized, the lyophilized formulation being combined with the sterile solution prior to administration. The compositions may contain pharmaceutically acceptable auxiliary substances as required to approximate physiological conditions, such as pH adjusting and buffering agents, tonicity adjusting agents and the like, for example, sodium acetate, sodium lactate, sodium chloride, potassium chloride, calcium chloride and stabilizing agents (e.g., 1-20% maltose and the like).

The pharmaceutical composition according to the invention may also be a bead system for delivering a therapeutic agent to a target cell. For example, a pectin/zein hydrogel bead system may be used to deliver neuregulin-4 or a pharmaceutical equivalent, analog, derivative or salt thereof to a target cell in a subject (Yan F et al, J Clin invest.2011.6 months; 121 (6): 2242-53).

Kit of the invention

In various embodiments, the present invention provides kits for treating a disorder, preventing a disorder, reducing the severity of a disorder, and/or slowing the progression of a disorder in a subject. The kit consists of or consists essentially of or comprises: an amount of a dual inhibitor of HDAC and GSK3 β; and instructions for using the dual inhibitor to treat, prevent, reduce the likelihood of, reduce the severity of, and/or slow the progression of a disorder in a subject.

In some embodiments, the dual inhibitor is a compound of formula I, formula II, formula III, formula I-1a, formula I-1b, formula I-1c, formula I-2, formula I-3, formula II-1, formula III-1, formula IIIb-1, formula IV, formula V, formula VI, formula VII, or a combination thereof.

In various embodiments, the dual inhibitor is conjugated to a particle. In various further embodiments, the dual inhibitor is attached to a cleavable enzyme substrate, and the cleavable enzyme substrate is attached to the magnetic particle.

In various embodiments, the present invention provides kits for treating a disorder, preventing a disorder, reducing the severity of a disorder, and/or slowing the progression of a disorder in a subject. The kit consists of or consists essentially of or comprises: an amount of an HDAC inhibitor; an amount of a GSK3 β inhibitor; and instructions for treating, preventing, reducing the likelihood of, reducing the severity of, and/or slowing the progression of a disorder in a subject using the HDAC inhibitor and GSK3 β inhibitor.

In various embodiments, the HDAC inhibitor is SAHA, TSA, TPX, MS-275, valproic acid, or CHAP31, or a functional equivalent, analog, derivative, or salt thereof, or a combination thereof. In various embodiments, the GSK3 β inhibitor is SB216763, TDZD-8, or Tideglusib (NP-12), or a functional equivalent, analog, derivative, or salt thereof, or a combination thereof. In various embodiments, the HDAC inhibitor and/or GSK3 β inhibitor is attached to a cleavable enzyme substrate, and the cleavable enzyme substrate is attached to a magnetic particle.

In various additional embodiments, a kit according to the invention further comprises a chemotherapeutic agent and instructions for using the chemotherapeutic agent to treat, prevent, reduce the likelihood of, reduce the severity of, and/or slow the progression of a disorder in a subject. In some embodiments, the chemotherapeutic agent in the kit is selected from the group consisting of actinomycin, alitretinol, all-trans retinoic acid, azacitidine, azathioprine, bevacizumab, bexarotene, bleomycin, bortezomib, carboplatin, capecitabine, cetuximab, cisplatin, chlorambucil, cyclophosphamide, cytarabine, daunorubicin, docetaxel, doxifluridine, doxorubicin, epirubicin, epothilone, erlotinib, etoposide, fluorouracil, gefitinib, gemcitabine, hydroxyurea, idarubicin, imatinib, damazepril, irinotecan, mechlorethamine, melphalan, mercaptopurine, methotrexate, mitoxantrone, omab, ofatumumab, oxaliplatin, paclitaxel, panitumumab, pemetrexed, rituximab, tafenoquine, teniposide, thioguanine, topotecan, bleomycin, and bortezomib, Tretinoin, valrubicin, vemurafenib, vinblastine, vincristine, vindesine, vinorelbine, vorinostat, romidepsin, 5-fluorouracil (5-FU), 6-mercaptopurine (6-MP), cladribine, clofarabine, floxuridine, fludarabine, pentostatin, mitomycin, ixabepilone, estramustine, prednisone, methylprednisolone, dexamethasone, or combinations thereof.

A kit is a collection of materials or components comprising at least one composition of the invention. The exact nature of the components constructed in the kit of the invention will depend on their intended purpose. In one embodiment, the kit is specifically configured for the purpose of treating a mammalian subject. In another embodiment, the kit is specifically configured for the purpose of treating a human subject. In still other embodiments, the kit is configured for veterinary use to treat a subject such as, but not limited to, farm animals, livestock, and laboratory animals.

Instructions for use may be included in the kit. "instructions for use" generally include a specific expression describing the technique used when the kit's components are used to produce the desired result. Optionally, the kit also contains other useful components, such as spray bottles or canisters, diluents, buffers, pharmaceutically acceptable carriers, syringes, catheters, applicators (e.g., applicators for creams, gels, lotions, or the like), pipetting or measuring instruments, dressing materials, or other useful implements, as can be readily identified by one of skill in the art.

The materials or components assembled in the kit can be provided to the practitioner and stored in any convenient and suitable manner that maintains their operability and usefulness. For example, the pharmaceutical composition may be in dissolved, dehydrated or lyophilized form; it may be provided at room temperature, refrigerated or frozen temperatures. The components are typically contained in a suitable packaging material. As used herein, the phrase "packaging material" refers to one or more physical structures used to contain the contents of a kit (e.g., a composition of the invention, etc.). The packaging material is constructed by well-known methods, preferably to provide a sterile, contaminant-free environment. As used herein, the term "package" refers to a suitable solid substrate or material, such as glass, plastic, paper, foil, and the like, capable of holding the components of a kit. Thus, for example, the package can be a glass vial for containing an appropriate amount of the composition described herein. The packaging material typically has an external label which indicates the contents and/or purpose of the kit and/or its components.

Exemplary embodiments of the aspects disclosed herein may be described by one or more of the following numbered paragraphs:

1. a compound of formula (IV):

wherein:

L₁and L₂Independently a linker;

R¹is an aromatic moiety, alkyl, acyl, cyclyl or heterocyclyl, each of which may be optionally substituted;

R²is hydrogen, lower alkyl, cyclyl, heterocyclyl, aryl or heteroaryl, each of which may be optionally substituted;

R³is absent or is an aromatic moiety, which may be optionally substituted;

p is 0, 1, 2, 3, 4, 5, 6,7, 8, 9 or 10; and is

wherein-L₁R¹To one nitrogen of the thiadiazolidine ring, and- (CH)₂)_p-R³-L₂-C(O)NHOR²Another nitrogen attached to the thiadiazolidine ring.

2. A compound of paragraph 1 having the structure of formula (VI):

3. a compound of paragraph 1 or 2 having the structure of formula (I):

wherein:

x is a linker group; and is

Y is absent or an aromatic substituent.

4. The compound of any one of paragraphs 1-3, having the structure of formula (I-1):

wherein n is an integer from 1 to 12.

5. The compound of any one of paragraphs 1-4, wherein the compound is:

6. a compound of paragraph 2 having the structure of formula (VI):

7. a compound of any of paragraphs 1, 2 or 6, having the structure of formula (II):

wherein:

x is a linker group and R is-L₁R¹。

8. The compound of any of paragraphs 1, 2, 6 or 7, having the structure of formula (II-1):

9. a compound of formula (V):

wherein:

L₁and L₂Independently a linker;

R¹is an aromatic moiety, alkyl, acyl, cyclyl or heterocyclyl, each of which may be optionally substituted;

R²is hydrogen, lower alkyl, cyclyl, heterocyclyl, aryl or heteroaryl, each of which may be optionally substituted;

R³is absent or is an aromatic moiety, which may be optionally substituted; and is

p is 0, 1, 2, 3, 4, 5, 6,7, 8, 9 or 10.

10. A compound of paragraph 9 having the structure of formula (III):

wherein:

x is a linker group; and is

Y is absent or an aromatic substituent.

11. The compound of paragraph 9 or 10, having the structure of formula (III-1):

12. a compound of paragraph 9 having the structure of formula (IIIb):

wherein:

x is a linker group; and is

Y is absent or an aromatic substituent.

13. The compound of paragraph 9 or 12, wherein the compound is of formula (IIIb-1):

14. the compound of any of paragraphs 1-13, wherein the compound is attached to a particle.

15. The compound of paragraph 14, wherein the particles are magnetic particles.

16. The compound of paragraph 14 or 15, wherein the compound is attached to the particle via a linker comprising a cleavable linker.

17. The compound of paragraph 16, wherein the cleavable linker is cleaved by an enzyme.

18. The compound of paragraph 16 or 17, wherein the cleavable linker is cleaved by an enzyme enriched in the cancer or tumor.

19. The compound of any of paragraphs 16-18, wherein the cleavable linker is cleaved by a peptidase enzyme enriched in cancer or tumor.

20. The compound of any of paragraphs 16-19, wherein the cleavable linker is a cleavable substrate of cathepsin G.

21. A composition comprising a dual inhibitor of HDAC and GSK3 β.

22. The composition of paragraph 21, wherein the dual inhibitor is a compound of any one of paragraphs 1-20.

23. The composition of paragraph 21 or 22, further comprising a pharmaceutically acceptable carrier or excipient.

24. The composition of any of paragraphs 21-23, wherein the composition is formulated for topical, intravascular, intravenous, intraarterial, intratumoral, intramuscular, subcutaneous, intraperitoneal, intranasal, or oral administration.

25. The composition of any of paragraphs 21-24, wherein the composition further comprises an anti-cancer therapeutic.

26. The composition of paragraph 25, wherein the anti-cancer therapeutic agent is a chemotherapeutic agent.

27. A method of treating, preventing, reducing the likelihood of, reducing the severity of, and/or slowing the progression of a disorder in a subject, comprising:

administering to the subject a therapeutically effective amount of a dual inhibitor of HDAC and GSK3 β, thereby treating, preventing, reducing the likelihood of, reducing the severity of, and/or slowing the progression of the disorder in the subject.

28. The method of paragraph 27, wherein the disorder is cancer or a tumor.

29. The method of paragraph 27 or 28, wherein the disorder is pancreatic cancer.

30. The method of any one of paragraphs 27-29, wherein the subject is a human.

31. The method of any of paragraphs 27-30, wherein the dual inhibitor is the compound of any of paragraphs 1-20.

32. The method of any of paragraphs 27-32, wherein the dual inhibitor is administered topically, intravascularly, intravenously, intraarterially, intratumorally, intramuscularly, subcutaneously, intraperitoneally, intranasally, or orally.

33. The method of any of paragraphs 27-32, wherein the dual inhibitor is administered as follows: about 0.001-0.01, 0.01-0.1, 0.1-0.5, 0.5-5, 5-10, 10-20, 20-50, 50-100, 100-200, 200-300, 300-400, 400-500, 500-600, 600-700, 700-800, 800-900 or 900-1000mg/kg or a combination thereof.

34. The method of any of paragraphs 27-32, wherein the dual inhibitor is administered as follows: about 0.001-0.01, 0.01-0.1, 0.1-0.5, 0.5-5, 5-10, 10-20, 20-50, 50-100, 100-200, 200-300, 300-400, 400-500, 500-600, 600-700, 700-800, 800-900 or 900-1000 μ g/kg or a combination thereof.

35. The method of any of paragraphs 27-34, wherein the dual inhibitor is administered about 1-3 times per day, 1-7 times per week, or 1-9 times per month.

36. The method of any one of paragraphs 27 to 35, wherein the dual inhibitor is administered for about 1-10 days, 10-20 days, 20-30 days, 30-40 days, 40-50 days, 50-60 days, 60-70 days, 70-80 days, 80-90 days, 90-100 days, 1-6 months, 6-12 months, or 1-5 years.

37. The method of any one of paragraphs 27-3627, further comprising administering an additional anti-cancer therapy.

38. The method of paragraph 37, wherein the dual inhibitor and the additional anti-cancer therapy are administered simultaneously or sequentially.

39. The method of paragraph 37 or 38, wherein the dual inhibitor is administered before, during or after the additional anti-cancer therapy is administered.

40. The method of any of paragraphs 37-39, wherein the additional anti-cancer therapy is selected from the group consisting of surgery, radiation therapy (radiotherapy), biological therapy, immunotherapy, chemotherapy, and any combination thereof.

41. The method of any one of paragraphs 37-40, wherein the additional anti-cancer therapy comprises administering an anti-cancer therapeutic to the subject.

42. The method of paragraph 41, wherein said dual inhibitor and said anti-cancer therapeutic are provided in one composition.

43. The method of paragraphs 41 or 42, wherein the dual inhibitor and the anti-cancer therapeutic agent are provided in separate compositions.

44. The method of any of paragraphs 41-43, wherein the anti-cancer therapeutic agent is a chemotherapeutic agent.

45. The method of any of paragraphs 27-44, wherein the dual inhibitor is attached to a magnetic particle, and the method further comprises directing the dual inhibitor to the cancer or tumor using a magnetic field.

46. A kit for treating, preventing, reducing the likelihood of, reducing the severity of, and/or slowing the progression of a disorder in a subject, comprising:

dual inhibitors of HDAC and GSK3 β; and

instructions for treating a disorder, preventing a disorder, reducing the likelihood of developing a disorder, reducing the severity of a disorder, and/or slowing the progression of a disorder in said subject using said dual inhibitor.

47. The kit of paragraph 46, wherein the dual inhibitor of HDAC and GSK3 β is a compound of any one of paragraphs 1-20.

48. The kit of paragraph 46 or 47, further comprising an anti-cancer therapeutic.

49. The kit of paragraph 48, wherein the anti-cancer agent is a chemotherapeutic agent.

50. A composition comprising an HDAC inhibitor and a GSK3 β inhibitor.

51. The composition of paragraph 50, wherein said HDAC inhibitor is selected from SAHA, TSA, TPX, MS-275, valproic acid or CHAP31 or a functional equivalent, analog, derivative or salt thereof, or a combination thereof.

52. The composition of paragraph 50 or 51, wherein the GSK3 β inhibitor is selected from SB216763, TDZD-8 or Tideglusib (NP-12), or a functional equivalent, analog, derivative or salt thereof, and any combination thereof.

53. The composition of any of paragraphs 50-52, wherein the HDAC inhibitor and/or the GSK3 β inhibitor is about 0.001-0.01, 0.01-0.1, 0.1-0.5, 0.5-5, 5-10, 10-20, 20-50, 50-100, 100-200, 200-300, 300-400, 400-500, 500-600, 600-700, 700-800, 800-900 or 900-1000mg/kg or a combination thereof.

54. The composition of any of paragraphs 50-53, wherein the HDAC inhibitor and/or the GSK3 β inhibitor is about 0.001-0.01, 0.01-0.1, 0.1-0.5, 0.5-5, 5-10, 10-20, 20-50, 50-100, 100-200, 200-300, 300-400, 400-500, 500-600, 600-700, 700-800, 800-900 or 900-1000 μ g/kg or a combination thereof.

55. The composition of any one of paragraphs 50-54, further comprising a pharmaceutically acceptable excipient or carrier.

56. The composition of any of paragraphs 50-55, wherein the composition is formulated for topical, intravascular, intravenous, intraarterial, intratumoral, intramuscular, subcutaneous, intraperitoneal, intranasal, or oral administration.

57. The composition of any of paragraphs 50-56, wherein the composition further comprises an anti-cancer therapeutic agent.

58. The composition of paragraph 57, wherein the anti-cancer therapeutic agent is a chemotherapeutic agent.

59. The composition of paragraph 57 or 58, wherein at least one of the HDAC inhibitor and the GSK3 β is conjugated to a particle.

60. The composition of paragraph 59, wherein the particles are magnetic particles.

61. The composition of paragraph 59 or 60, wherein the HDAC inhibitor and/or GSK3 β is attached to the particle through a linker comprising a cleavable linker.

62. The composition of paragraph 61, wherein the cleavable linker is cleaved by an enzyme.

63. The composition of paragraph 61 or 62, wherein the cleavable linker is cleaved by an enzyme enriched in the cancer or tumor.

64. The composition of any one of paragraphs 61-63, wherein the cleavable linker is cleaved by a peptidase enriched in cancer or tumor.

65. The composition of any one of paragraphs 61-64, wherein the cleavable linker is a cleavable substrate of cathepsin G.

66. A method of treating, preventing, reducing the likelihood of, reducing the severity of, and/or slowing the progression of a disorder in a subject, comprising:

administering to the subject a therapeutically effective amount of an HDAC inhibitor and a GSK3 β inhibitor, thereby treating, preventing, reducing the likelihood of, reducing the severity of, and/or slowing the progression of the disorder in the subject.

67. The method of paragraph 66, wherein the disorder is cancer or a tumor.

68. The method of paragraph 66 or 67, wherein the disorder is pancreatic cancer.

69. The method of any one of paragraphs 66-68, wherein the subject is a human.

70. The method of any of paragraphs 66-69, wherein the HDAC inhibitor and the GSK3 β inhibitor are provided in one composition.

71. The method of any of paragraphs 66-69, wherein the HDAC inhibitor and the GSK3 β inhibitor are provided in separate compositions.

72. The method of any of paragraphs 66-71, wherein the HDAC inhibitor and the GSK3 β inhibitor are administered simultaneously or sequentially.

73. The method of any of paragraphs 66-72, wherein the HDAC inhibitor is administered before, during, or after the administration of the GSK3 β inhibitor.

74. The method of any of paragraphs 66-73, wherein the HDAC inhibitor is SAHA, TSA, TPX, MS-275, valproic acid, or CHAP31 or a functional equivalent, analog, derivative, or salt thereof, or a combination thereof.

75. The method of any one of paragraphs 66-74, wherein the GSK3 β inhibitor is SB216763, TDZD-8, or Tideglusib (NP-12), or a functional equivalent, analog, derivative, or salt thereof, or a combination thereof.

76. The method of any of paragraphs 66-75, wherein the HDAC inhibitor and/or the GSK3 β inhibitor is administered topically, intravascularly, intravenously, intraarterially, intratumorally, intramuscularly, subcutaneously, intraperitoneally, intranasally, or orally.

77. The method of any of paragraphs 66-76, wherein the HDAC inhibitor and/or the GSK3 β inhibitor is administered as follows: about 0.001-0.01, 0.01-0.1, 0.1-0.5, 0.5-5, 5-10, 10-20, 20-50, 50-100, 100-200, 200-300, 300-400, 400-500, 500-600, 600-700, 700-800, 800-900 or 900-1000mg/kg or a combination thereof.

78. The method of any of paragraphs 66-76, wherein the HDAC inhibitor and/or the GSK3 β inhibitor is administered as follows: about 0.001-0.01, 0.01-0.1, 0.1-0.5, 0.5-5, 5-10, 10-20, 20-50, 50-100, 100-200, 200-300, 300-400, 400-500, 500-600, 600-700, 700-800, 800-900 or 900-1000 μ g/kg or a combination thereof.

79. The method of any of paragraphs 66-78, wherein the HDAC inhibitor and/or the GSK3 β inhibitor is administered about 1-3 times per day, 1-7 times per week, or 1-9 times per month.

80. The method of any of paragraphs 66-79, wherein the HDAC inhibitor and/or the GSK3 β inhibitor is administered for about 1-10 days, 10-20 days, 20-30 days, 30-40 days, 40-50 days, 50-60 days, 60-70 days, 70-80 days, 80-90 days, 90-100 days, 1-6 months, 6-12 months, or 1-5 years.

81. The method of any one of paragraphs 66 to 8027, further comprising administering an additional anti-cancer therapy.

82. The method of paragraph 81, wherein said HDAC inhibitor, said GSK3 β inhibitor and said additional anti-cancer therapy are administered simultaneously or sequentially.

83. The method of paragraph 81 or 82, wherein said HDAC inhibitor and/or said GSK3 β inhibitor is administered before, during or after administration of said additional anti-cancer therapy.

84. The method of any of paragraphs 81-83, wherein the additional anti-cancer therapy is selected from the group consisting of surgery, radiation therapy (radiotherapy), biological therapy, immunotherapy, chemotherapy, and any combination thereof.

85. The method of any one of paragraphs 81-84, wherein the additional anti-cancer therapy comprises administering an anti-cancer therapeutic to the subject.

86. The method of any of paragraphs 81-85, wherein the HDAC inhibitor, the GSK3 β inhibitor and the chemotherapeutic agent are provided in separate compositions.

87. The method of any of paragraphs 81-85, wherein at least two of the HDAC inhibitor, the GSK3 β inhibitor and the anti-cancer therapeutic are provided in one composition.

88. The method of any of paragraphs 81-85, wherein all three of the HDAC inhibitor, the GSK3 β inhibitor and the anti-cancer therapeutic are provided in one composition.

89. The method of any of paragraphs 81-88, wherein the anti-cancer therapeutic agent is a chemotherapeutic agent.

90. The method of paragraph 66, wherein at least one of the HDAC inhibitor and GSK3 β is conjugated to a particle.

91. The method of paragraph 90, wherein the particles are magnetic particles.

92. The method of paragraph 90 or 91, wherein the HDAC inhibitor and/or GSK3 β is attached to the particle through a linker comprising a cleavable linker.

93. The method of paragraph 92, wherein the cleavable linker is cleaved by an enzyme.

94. The method of paragraph 92 or 93, wherein the cleavable linker is cleaved by an enzyme enriched in the cancer or tumor.

95. The method of any of paragraphs 92-94, wherein the cleavable linker is cleaved by a peptidase enriched in the cancer or tumor.

96. The method of any one of paragraphs 92-95, wherein the cleavable linker is a cleavable substrate of cathepsin G.

97. The method of any of paragraphs 92-96, wherein at least one of the HDAC inhibitor and the GSK3 β is attached to a magnetic particle, and the method further comprises directing the HDAC inhibitor and/or the GSK3 β to a cancer or tumor using a magnetic field.

98. A kit for treating, preventing, reducing the likelihood of, reducing the severity of, and/or slowing the progression of a disorder in a subject, comprising:

(ii) an HDAC inhibitor;

GSK3 β inhibitors; and

instructions for treating, preventing, reducing the likelihood of, reducing the severity of, and/or slowing the progression of a disorder in said subject using said HDAC inhibitor and said GSK3 β inhibitor.

99. The kit of paragraph 98, further comprising an anti-cancer therapeutic.

100. The kit of paragraph 98 or 99, wherein the anti-cancer agent is a chemotherapeutic agent.

101. The kit of any of paragraphs 98-101, wherein at least one of the HDAC inhibitor and the GSK3 β is conjugated to a particle.

102. The kit of paragraph 101, wherein the particles are magnetic particles.

103. The kit of paragraph 101 or 102, wherein the HDAC inhibitor and/or GSK3 β is linked to the particle through a linker comprising a cleavable linker.

104. The kit of paragraph 103, wherein the cleavable linker is cleaved by an enzyme.

105. The kit of paragraph 103 or 104, wherein the cleavable linker is cleaved by an enzyme enriched in the cancer or tumor.

106. The kit of any one of paragraphs 103-105, wherein the cleavable linker is cleaved by a peptidase enzyme that is enriched in cancer or tumor.

107. The kit of any one of paragraphs 103-106, wherein the cleavable linker is a cleavable substrate for cathepsin G.

Many variations and alternative elements have been disclosed in embodiments of the invention. Further variations and alternative elements will be apparent to those skilled in the art. Among these variations are, without limitation, the selection of the building blocks of the compositions of the invention, and the diseases and other clinical conditions that can be diagnosed, predicted or treated therewith. Embodiments of the invention may specifically include or exclude any of these variations or elements.

In some embodiments, numerical values used to describe and suggest quantities of ingredients, properties (e.g., concentrations), reaction conditions, and the like of certain embodiments of the present invention are understood to be modified in some instances by the term "about". Accordingly, in some embodiments, the numerical parameters set forth in the written description and attached claims are approximations that may vary depending upon the particular properties sought to be obtained by a particular embodiment. In some embodiments, numerical parameters should be construed in light of the number of reported significant digits and by applying ordinary rounding techniques. Notwithstanding that the numerical ranges and parameters setting forth the broad scope of some embodiments of the invention are approximations, the numerical values set forth in the specific examples are reported as precisely as possible. The numerical ranges set forth in some embodiments of the invention may include some necessary error from the standard deviation found in their respective test measurements.

Groupings of alternative elements or embodiments of the invention disclosed herein are not to be construed as limiting. Each group member may be referred to or suggested individually or in any combination with other elements from the group or other elements present therein. For convenience and/or patentability, one or more members of a group may be included in or deleted from the group. When any such inclusion or deletion occurs, the specification herein is considered to include the group so modified as to satisfy the written description of markush group as used in the appended claims.

Examples

The invention will be further explained by the following examples, which are intended to be purely exemplary of the invention and should not be construed as limiting the invention in any way. The following examples are provided to better illustrate the claimed invention and should not be construed as limiting the scope of the invention. To the extent that specific materials are mentioned, they are for illustrative purposes only, and are not intended to limit the invention. Those skilled in the art will be able to develop equivalent means or reactants without the exercise of inventive skill and without departing from the scope of the present invention.

Example 1

Treatment of pancreatic cancer by novel compounds and methods that simultaneously inhibit growth-promoting GSK beta and metastasis and treatment-resistance-promoting HDAC

In various embodiments, the present invention provides cancer treatments that combine inhibitors of both GSK3 β and HDACs for K-ras-mediated neoplasms. The general scheme is shown in fig. 1, and the experimental results are shown in fig. 2-7.

The inventors used a mouse model of pancreatic cancer in which the oncogene call mutant K-ras (pdx1-Cre-LSL-Kras) was expressed in the pancreas. These mice were exposed to cigarette smoke for 6 weeks in the smoke chamber. Groups of mice were injected with the GSK3 β inhibitor TDZD-8(4mg/Kg, 3 times per week) and/or the HDAC inhibitor Saha (50mg/Kg, 5 times per week) for 6 weeks.

The present inventors found that animals receiving Saha had significantly reduced early stage cancer lesions known as pancreatic intraepithelial neoplasia (PanIN) compared to the control group; and the combination of Saha and TDZD-8 significantly improved this effect compared to each individual compound (figure 2). The same effect was observed when fibrosis was measured with collagen staining. Fibrosis is a measure of cancer activity. Indeed, the combination of Saha and TDZD-8 synergistically reduced fibrosis (figure 3).

Inhibition of GSK3 β induced upregulation of EMT as shown by measurement of vimentin, a well-established measure for EMT (figure 4). The HDAC inhibitor Saha prevented this effect of GSK3 β (fig. 4). Saha also inhibits transcription factors known to regulate EMT, such as Twist and Snail. The combination of both inhibitors with gemcitabine, a chemotherapeutic agent for pancreatic cancer, resulted in complete inhibition of the EMT marker vimentin and its transcription factor (fig. 4).

The combination of low doses of inhibitors induced a synergistic effect on cancer cell survival (fig. 5-7). And more importantly, HDAC inhibitors prevent the EMT/metastasis promoting effect of GSK3 β inhibition, thereby causing dual beneficial effects by a synergistic effect on cell survival and growth and by reversing the carcinogenic side effects of one inhibitor.

Example 2: in vitro and in vivo studies of exemplary Dual inhibitors

Effect of ALB-185357 on cell survival

BxPC-3 pancreatic cancer cell line 72 was cultured in the presence or absence of different doses of a combination of saha and tidegusib or ALB-185357 and cell survival was measured by MTT assay. The data in figure 13 show that compound ALB-185357 dose-dependently reduced cell survival. ALB-185357 was more potent than the combination of the HDAC inhibitor saha and the GSK-3 β inhibitor tideglusib. Significance was achieved when 300nM ALB-185357 was used and its effect on cell survival was greater than the effect of the combination of saha and tideglusib. As can be seen from fig. 4 and 7, the combination of saha and tideglusib has a synergistic effect on cell death, proliferation and EMT measurements. Thus, the effect of ALB-185357 (a dual inhibitor of HDAC and GSK-3 β) represents an additional synergistic effect on the effects observed with the combination of the individual agents.

Effect of ALB-185357 on apoptosis

MIA PaCa-2 cells 72 were cultured in the presence or absence of different doses of ALB-185357 and apoptosis was assessed by measuring DNA fragmentation. The results are shown in fig. 14. As can be seen from the data in fig. 14, ALB-185357 dose-dependently increased apoptosis as measured by the level of DNA fragmentation, and significance was achieved at a dose of 300 nM.

Effect of ALB-185357 and Gemcitabine on apoptosis

MIAPaCa-2 cells 72 were cultured in the presence or absence of different doses of ALB-185357 or 1ng/ml of low dose gemcitabine and apoptosis was assessed by measuring DNA fragmentation. The data in figure 15 show that the combination of ALB-185357 and gemcitabine has a stronger effect on inducing apoptosis than either drug alone or the expected additive effect.

Effect of ALB-188540 and ALB-185643 on cell survival

BxPC-3 pancreatic cancer cells 72 were cultured in the presence or absence of different doses of ALB-188540 or ALB-185643 and cell survival was measured by MTT assay. The data in fig. 16 and 16B indicate that compounds ALB-188540 (fig. 16A) and ALB-185643 (fig. 16B) have similar effects on the survival of BxPC3 cells as compound ALB-185357.

Effect of ALB-185357 on cell survival of different cancer and non-cancer cell types

Cells 72 were cultured in the presence or absence of different doses of ALB-185357 and cell survival was measured by MTT assay (fig. 17 and 18) or by counting cell numbers (fig. 19). The results of the MTT assay are shown in fig. 17 and 18, and the results of cell counting are shown in fig. 19. This data indicates that ALB-185357 inhibits cell survival of multiple cancer cell types.

Cells 72 were cultured in the presence or absence of different doses of ALB-185357 and cell survival was measured by MTT assay. The data in figure 20 show that compound ALB-185357 did not affect cell survival of normal pancreatic ductal cells.

In summary, the data in FIGS. 17-20 demonstrate that ALB-185357 is highly toxic to cancer cells from different cancer types, but not to normal cells.

Effect of ALB-185357 on histone acetylation and GSK-3 beta phosphorylation/inhibition

Cells 72 were cultured in the presence or absence of different doses of ALB-185357 and protein levels were measured by Western. The data indicate that pathways predicted to be modulated by ALB-185357 are indeed modulated by drugs. As seen in figure 21, compound ALB-185357 dose-dependently upregulated the predicted targets in the MIA PaCa-2 pancreatic cancer cell line, namely target histone acetylation and GSK-3 β phosphorylation/inhibition.

Effect of ALB-185357 on histone acetylation and GSK-3 beta phosphorylation/inhibition

Cells 72 were cultured in the presence or absence of different doses of ALB-185357 and protein level maps were measured by Western (22A) and cell infiltration was measured in a Matrigel infiltration Chamber (Matrigel Invasion Chamber) (fig. 22B). The data in fig. 22A indicate that ALB-185357 down-regulates proteins that mediate cell metastasis and therapy resistance. The data in fig. 22B indicate that the ability of cancer cells to infiltrate is down-regulated.

Effect of ALB-185357 on survival in vivo

KPC mice with Kras and p53 mutations and spontaneously developing pancreatic cancer were ip-injected with 5mg/Kg ALBl85357 three times a week from 8 weeks of age until death. As seen in figure 23, compound ALB-185357 improved survival in animals with advanced pancreatic cancer.

The various methods and techniques described above provide a number of ways to implement the present application. Of course, it is to be understood that not necessarily all objectives or advantages described may be achieved in accordance with any particular embodiment described herein. Thus, for example, those skilled in the art will recognize that the methods may be practiced in a manner that achieves or optimizes one advantage or group of advantages as taught herein without necessarily achieving other objectives or advantages as may be taught or suggested herein. Various alternatives are mentioned herein. It will be understood that some preferred embodiments specifically include one, another or more features while others specifically exclude one, another or more features, while still others mitigate a particular feature by including one, another or more advantageous features.

In addition, the skilled person will recognize the applicability of various features from different embodiments. Similarly, the various elements, features and steps discussed above, as well as other known equivalents for these elements, features or steps, can be used in various combinations by one of ordinary skill in the art to implement methods in accordance with the principles described herein. Among the various elements, features and steps, some will be specifically included and others specifically excluded in various embodiments.

Although the present application is disclosed in the context of certain embodiments and examples, it will be understood by those skilled in the art that the embodiments of the present application extend beyond the specifically disclosed embodiments to other alternative embodiments and/or uses and modifications and equivalents.

Preferred embodiments of this application are described herein, including the best mode known to the inventors for carrying out the application. Variations of those preferred embodiments may become apparent to those of ordinary skill in the art upon reading the foregoing description. It is contemplated that one skilled in the art may employ such variations as desired, and practice the present application in other ways than those specifically described. Accordingly, many embodiments of the present application include all modifications and equivalents of the subject matter recited in the claims appended hereto as permitted by applicable law. Moreover, any combination of the above-described elements in all possible variations thereof is encompassed by the application unless otherwise indicated herein or otherwise clearly contradicted by context.

All patents, patent applications, publications and other materials cited herein, such as articles, books, specifications, publications, documents, things, and/or the like, are hereby incorporated by reference in their entirety for all purposes, except to the extent any litigation history associated therewith is inconsistent or contrary to this document, or anywhere in the art that has a limiting effect on the broad scope of the present and subsequent claims associated with the document. For example, if the description, definitions, and/or terminology associated with any incorporated material is not in any way inconsistent with those associated with this document, the description, definitions, and/or terminology used in this document shall govern.

It is to be understood that the embodiments of the application disclosed herein are illustrative of the principles of the embodiments of the present application. Other modifications that may be employed may be within the scope of the present application. Thus, for example, but not limiting of, alternative configurations of the embodiments of the present application may be used in accordance with the teachings herein. Thus, embodiments of the present application are not limited to what has been precisely shown and described.

Various embodiments of the present invention are described in the above detailed description of the invention. While these descriptions directly describe the above embodiments, it is to be understood that modifications and/or variations of the specific embodiments shown and described herein may be envisioned by those skilled in the art. Any such modifications and variations that fall within the scope of this specification are intended to be included herein as well. Unless otherwise indicated, it is the intention of the inventors that the words and phrases in the specification and claims be given the ordinary and customary meaning to those skilled in the art to which this application pertains.

The foregoing descriptions of various embodiments of the present invention known to the applicant have been presented at the time of filing the application and are intended for purposes of illustration and description. This application is not intended to be exhaustive or to limit the invention to the precise form disclosed, and many modifications and variations are possible in light of the above teaching. The described embodiments are intended to explain the principles of the invention and its practical application and to enable others skilled in the art to utilize the invention in various embodiments and with various modifications as are suited to the particular use contemplated. Therefore, it is intended that the invention not be limited to the particular embodiments disclosed for carrying out this invention.

While particular embodiments of the present invention have been shown and described, it will be obvious to those skilled in the art that, based upon the teachings herein, changes and modifications may be made without departing from this invention and its broader aspects and, therefore, the appended claims are to encompass within their scope all such changes and modifications as are within the true spirit and scope of this invention.

Claims (107)

1. A compound of formula (IV):

wherein:

L₁and L₂Independently a linker;

R¹is an aromatic moiety, alkyl, acyl, cyclyl or heterocyclyl, each of which may be optionally substituted;

R²is hydrogen,Lower alkyl, cyclyl, heterocyclyl, aryl or heteroaryl, each of which may be optionally substituted;

R³is absent or is an aromatic moiety, which may be optionally substituted;

p is 0, 1, 2, 3, 4, 5, 6,7, 8, 9 or 10; and is

wherein-L₁R¹To one nitrogen of the thiadiazolidine ring, and- (CH)₂)_p-R³-L₂-C(O)NHOR²Another nitrogen attached to the thiadiazolidine ring.

2. The compound of claim 1, having the structure of formula (VI):

3. the compound of claim 2, having the structure of formula (I):

wherein:

x is a linker group; and is

Y is absent or an aromatic substituent.

4. The compound of claim 3, having the structure of formula (I-1):

wherein n is an integer from 1 to 12.

5. The compound of claim 4, wherein the compound is:

6. the compound of claim 2, having the structure of formula (VI):

7. the compound of claim 6, having the structure of formula (II):

wherein:

x is a linker group and R is-L₁R¹。

8. The compound of claim 7, having the structure of formula (II-1):

9. a compound of formula (V):

wherein:

L₁and L₂Independently a linker;

R¹is an aromatic moiety, alkyl, acyl, cyclyl or heterocyclyl, each of which may be optionally substituted;

R²is hydrogen, lower alkyl, cyclyl, heterocyclyl, aryl or heteroaryl, each of which may be optionally substituted;

R³is absent or is an aromatic moiety, which may be optionally substituted; and is

p is 0, 1, 2, 3, 4, 5, 6,7, 8, 9 or 10.

10. The compound of claim 9, having the structure of formula (III):

wherein:

x is a linker group; and is

Y is absent or an aromatic substituent.

11. The compound of claim 10, having the structure of formula (III-1):

12. the compound of claim 9, having the structure of formula (IIIb):

wherein:

x is a linker group; and is

Y is absent or an aromatic substituent.

13. The compound of claim 12, wherein the compound is of formula (IIIb-1):

14. the compound of any one of claims 1-13, wherein the compound is attached to a particle.

15. The compound of claim 14, wherein the particle is a magnetic particle.

16. The compound of claim 14 or 15, wherein the compound is attached to a particle via a linker comprising a cleavable linker.

17. The compound of claim 16, wherein the cleavable linker is cleaved by an enzyme.

18. The compound of claim 17, wherein the cleavable linker is cleaved by an enzyme enriched in cancer or tumor.

19. The compound of claim 18, wherein the cleavable linker is cleaved by a peptidase enzyme enriched in cancer or tumors.

20. The compound of claim 19, wherein the cleavable linker is a cleavable substrate of cathepsin G.

21. A composition comprising a dual inhibitor of HDAC and GSK3 β.

22. The composition of claim 21, wherein the dual inhibitor is a compound of any one of claims 1-20.

23. The composition of claim 21, further comprising a pharmaceutically acceptable carrier or excipient.

24. The composition of claim 21, wherein the composition is formulated for topical, intravascular, intravenous, intraarterial, intratumoral, intramuscular, subcutaneous, intraperitoneal, intranasal, or oral administration.

25. The composition of claim 21, wherein the composition further comprises an anti-cancer therapeutic agent.

26. The composition of claim 25, wherein the anti-cancer therapeutic is a chemotherapeutic agent.

27. A method of treating, preventing, reducing the likelihood of, reducing the severity of, and/or slowing the progression of a disorder in a subject, comprising:

administering to the subject a therapeutically effective amount of a dual inhibitor of HDAC and GSK3 β, thereby treating, preventing, reducing the likelihood of, reducing the severity of, and/or slowing the progression of the disorder in the subject.

28. The method of claim 27, wherein the disorder is cancer or a tumor.

29. The method of claim 28, wherein the disorder is pancreatic cancer.

30. The method of claim 27, wherein the subject is a human.

31. The method of claim 27, wherein the dual inhibitor is a compound of any one of claims 1-20.

32. The method of claim 27, wherein the dual inhibitor is administered topically, intravascularly, intravenously, intraarterially, intratumorally, intramuscularly, subcutaneously, intraperitoneally, intranasally, or orally.

33. The method of claim 27, wherein the dual inhibitor is administered as follows: about 0.001-0.01, 0.01-0.1, 0.1-0.5, 0.5-5, 5-10, 10-20, 20-50, 50-100, 100-200, 200-300, 300-400, 400-500, 500-600, 600-700, 700-800, 800-900 or 900-1000mg/kg or a combination thereof.

34. The method of claim 27, wherein the dual inhibitor is administered as follows: about 0.001-0.01, 0.01-0.1, 0.1-0.5, 0.5-5, 5-10, 10-20, 20-50, 50-100, 100-200, 200-300, 300-400, 400-500, 500-600, 600-700, 700-800, 800-900 or 900-1000 μ g/kg or a combination thereof.

35. The method of claim 27, wherein the dual inhibitor is administered at about 1-3 times per day, 1-7 times per week, or 1-9 times per month.

36. The method of claim 27, wherein the dual inhibitor is administered for about 1-10 days, 10-20 days, 20-30 days, 30-40 days, 40-50 days, 50-60 days, 60-70 days, 70-80 days, 80-90 days, 90-100 days, 1-6 months, 6-12 months, or 1-5 years.

37. The method of claim 2727, further comprising administering an additional anti-cancer therapy.

38. The method of claim 37, wherein the dual inhibitor and the additional anti-cancer therapy are administered simultaneously or sequentially.

39. The method of claim 37, wherein the dual inhibitor is administered before, during, or after the additional anti-cancer therapy is administered.

40. The method of claim 37, wherein the additional anti-cancer therapy is selected from the group consisting of surgery, radiation therapy (radiotherapy), biological therapy, immunotherapy, chemotherapy, and any combination thereof.

41. The method of claim 40, wherein the additional anti-cancer therapy comprises administering an anti-cancer therapeutic to the subject.

42. The method of claim 41, wherein the dual inhibitor and the anti-cancer therapeutic agent are provided in one composition.

43. The method of claim 41, wherein the dual inhibitor and the anti-cancer therapeutic agent are provided in different compositions.

44. The method of claim 41, wherein the anti-cancer therapeutic is a chemotherapeutic agent.

45. The method of claim 27, wherein the dual inhibitor is attached to a magnetic particle, and the method further comprises directing the dual inhibitor to a cancer or tumor using a magnetic field.

46. A kit for treating, preventing, reducing the likelihood of, reducing the severity of, and/or slowing the progression of a disorder in a subject, comprising:

dual inhibitors of HDAC and GSK3 β; and

instructions for treating, preventing, reducing the likelihood of, reducing the severity of, and/or slowing the progression of the disorder in the subject using the dual inhibitor.

47. The kit of claim 46, wherein the dual inhibitor of HDAC and GSK3 β is a compound according to any one of claims 1-20.

48. The kit of claim 46, further comprising an anti-cancer therapeutic.

49. The kit of claim 48, wherein the anti-cancer agent is a chemotherapeutic agent.

50. A composition comprising an HDAC inhibitor and a GSK3 β inhibitor.

51. The composition of claim 50, wherein the HDAC inhibitor is selected from the group consisting of SAHA, TSA, TPX, MS-275, valproic acid, or CHAP31, or a functional equivalent, analog, derivative, or salt thereof, and any combination thereof.

52. The composition of claim 50, wherein the GSK3 β inhibitor is selected from the group consisting of SB216763, TDZD-8 or Tideglusib (NP-12), or a functional equivalent, analog, derivative or salt thereof, and any combination thereof.

53. The composition of claim 50, wherein the HDAC inhibitor and/or the GSK3 β inhibitor is about 0.001-0.01, 0.01-0.1, 0.1-0.5, 0.5-5, 5-10, 10-20, 20-50, 50-100, 100-200, 200-300, 300-400, 400-500, 500-600, 600-700, 700-800, 800-900, or 900-1000mg/kg or a combination thereof.

54. The composition of claim 50, wherein the HDAC inhibitor and/or the GSK3 β inhibitor is about 0.001-0.01, 0.01-0.1, 0.1-0.5, 0.5-5, 5-10, 10-20, 20-50, 50-100, 100-200, 200-300, 300-400, 400-500, 500-600, 600-700, 700-800, 800-900, or 900-1000 μ g/kg or a combination thereof.

55. The composition of claim 50, further comprising a pharmaceutically acceptable excipient or carrier.

56. The composition of claim 50, wherein the composition is formulated for topical, intravascular, intravenous, intraarterial, intratumoral, intramuscular, subcutaneous, intraperitoneal, intranasal, or oral administration.

57. The composition of claim 50, wherein the composition further comprises an anti-cancer therapeutic agent.

58. The composition of claim 50, wherein the anti-cancer therapeutic agent is a chemotherapeutic agent.

59. The composition of claim 50, wherein at least one of the HDAC inhibitor and the GSK3 β is conjugated to a particle.

60. The composition of claim 59, wherein the particles are magnetic particles.

61. The composition of claim 59 or 60, wherein the HDAC inhibitor and/or the GSK3 β are linked to the particle via a linker comprising a cleavable linker.

62. The composition of claim 61, wherein said cleavable linker is cleaved by an enzyme.

63. The composition of claim 62, wherein the cleavable linker is cleaved by an enzyme enriched in a cancer or tumor.

64. The composition of claim 63, wherein the cleavable linker is cleaved by a peptidase enriched in cancer or tumor.

65. The composition of claim 64, wherein said cleavable linker is a cleavable substrate of cathepsin G.

66. A method of treating, preventing, reducing the likelihood of, reducing the severity of, and/or slowing the progression of a disorder in a subject, comprising:

administering to the subject a therapeutically effective amount of an HDAC inhibitor and a GSK3 β inhibitor, thereby treating, preventing, reducing the likelihood of, reducing the severity of, and/or slowing the progression of the disorder in the subject.

67. The method of claim 66, wherein the disorder is cancer or a tumor.

68. The method of claim 67, wherein the disorder is pancreatic cancer.

69. The method of claim 66, wherein the subject is a human.

70. The method of claim 66, wherein the HDAC inhibitor and the GSK3 β inhibitor are provided in one composition.

71. The method of claim 66, wherein the HDAC inhibitor and the GSK3 β inhibitor are provided in different compositions.

72. The method of claim 66, wherein the HDAC inhibitor and the GSK3 β inhibitor are administered simultaneously or sequentially.

73. The method of claim 66, wherein the HDAC inhibitor is administered before, during, or after the administration of the GSK3 β inhibitor.

74. The method of claim 66, wherein the HDAC inhibitor is SAHA, TSA, TPX, MS-275, valproic acid, or CHAP31, or a functional equivalent, analog, derivative, or salt thereof, or a combination thereof.

75. The method of claim 66, wherein the GSK3 β inhibitor is SB216763, TDZD-8 or Tideglusib (NP-12), or a functional equivalent, analog, derivative or salt thereof, or a combination thereof.

76. The method of claim 66, wherein the HDAC inhibitor and/or the GSK3 β inhibitor is administered topically, intravascularly, intravenously, intraarterially, intratumorally, intramuscularly, subcutaneously, intraperitoneally, intranasally, or orally.

77. The method of claim 66, wherein the HDAC inhibitor and/or the GSK3 β inhibitor is administered as follows: about 0.001-0.01, 0.01-0.1, 0.1-0.5, 0.5-5, 5-10, 10-20, 20-50, 50-100, 100-200, 200-300, 300-400, 400-500, 500-600, 600-700, 700-800, 800-900 or 900-1000mg/kg or a combination thereof.

78. The method of claim 66, wherein the HDAC inhibitor and/or the GSK3 β inhibitor is administered as follows: about 0.001-0.01, 0.01-0.1, 0.1-0.5, 0.5-5, 5-10, 10-20, 20-50, 50-100, 100-200, 200-300, 300-400, 400-500, 500-600, 600-700, 700-800, 800-900 or 900-1000 μ g/kg or a combination thereof.

79. The method of claim 66, wherein the HDAC inhibitor and/or the GSK3 β inhibitor is administered about 1-3 times per day, 1-7 times per week, or 1-9 times per month.

80. The method of claim 66, wherein the HDAC inhibitor and/or the GSK3 β inhibitor is administered for about 1-10 days, 10-20 days, 20-30 days, 30-40 days, 40-50 days, 50-60 days, 60-70 days, 70-80 days, 80-90 days, 90-100 days, 1-6 months, 6-12 months, or 1-5 years.

81. The method of claim 6627, further comprising administering an additional anti-cancer therapy.

82. The method of claim 81, wherein the HDAC inhibitor, the GSK3 β inhibitor and the additional anti-cancer therapy are administered simultaneously or sequentially.

83. The method of claim 81, wherein the HDAC inhibitor and/or the GSK3 β inhibitor is administered before, during, or after the additional anti-cancer therapy is administered.

84. The method of claim 81, wherein the additional anti-cancer therapy is selected from the group consisting of surgery, radiation therapy (radiotherapy), biological therapy, immunotherapy, chemotherapy, and any combination thereof.

85. The method of claim 84, wherein the additional anti-cancer therapy comprises administering an anti-cancer therapeutic to the subject.

86. The method of claim 85, wherein the HDAC inhibitor, the GSK3 β inhibitor, and the chemotherapeutic agent are provided in different compositions.

87. The method of claim 85, wherein at least two of the HDAC inhibitor, the GSK3 β inhibitor, and the anti-cancer therapeutic are provided in one composition.

88. The method of claim 87, wherein all three of the HDAC inhibitor, the GSK3 β inhibitor, and the anti-cancer therapeutic are provided in one composition.

89. The method of claim 85, wherein the anti-cancer therapeutic is a chemotherapeutic agent.

90. The method of claim 66, wherein at least one of the HDAC inhibitor and the GSK3 β is conjugated to a particle.

91. The method of claim 90, wherein the particles are magnetic particles.

92. The method of claim 90 or 91, wherein the HDAC inhibitor and/or the GSK3 β is linked to the particle via a linker comprising a cleavable linker.

93. The method of claim 92, wherein the cleavable linker is cleaved by an enzyme.

94. The method of claim 93, wherein the cleavable linker is cleaved by an enzyme enriched in a cancer or tumor.

95. The method of claim 94, wherein said cleavable linker is cleaved by a peptidase enriched in cancer or tumor.

96. The method of claim 95, wherein said cleavable linker is a cleavable substrate of cathepsin G.

97. The method of claim 68, wherein at least one of the HDAC inhibitor and GSK3 β is attached to a magnetic particle, and the method further comprises directing the HDAC inhibitor and/or the GSK3 β to a cancer or tumor using a magnetic field.

98. A kit for treating, preventing, reducing the likelihood of, reducing the severity of, and/or slowing the progression of a disorder in a subject, comprising:

(ii) an HDAC inhibitor;

GSK3 β inhibitors; and

instructions for treating, preventing, reducing the likelihood of, reducing the severity of, and/or slowing the progression of the disorder in the subject using the HDAC inhibitor and the GSK3 β inhibitor.

99. The kit of claim 98, further comprising an anti-cancer therapeutic.

100. The kit of claim 99, wherein the anti-cancer agent is a chemotherapeutic agent.

101. The kit of claim 98, wherein at least one of the HDAC inhibitor and GSK3 β is conjugated to a particle.

102. The kit of claim 101, wherein the particles are magnetic particles.

103. The kit of claim 101, wherein the HDAC inhibitor and/or GSK3 β is attached to the particle through a linker comprising a cleavable linker.

104. The kit of claim 102, wherein the cleavable linker is cleaved by an enzyme.

105. The kit of claim 103, wherein said cleavable linker is cleaved by an enzyme enriched in a cancer or tumor.

106. The kit of claim 104, wherein the cleavable linker is cleaved by a peptidase enzyme enriched in cancer or tumor.

107. The kit of claim 106, wherein said cleavable linker is a cleavable substrate of cathepsin G.