Treatment of Multiple Myeloma

Abstract

To provide a method for treating multiple myeloma.SOLUTION: Provided is a method comprising administering to an individual an effective amount of trichostatin A (TSA). Preferably, the effective amount is an amount sufficient to reduce expression of at least one gene in the individual, and at least one gene is selected from the group consisting of CCNB1, AURKB, CDC2, BIRC5, KIF11, KIF2C, TOP2A, ASPM, CKS1B, and WEE1.SELECTED DRAWING: None

Description

Cross - reference to related applications

[0001] This application claims the benefit of co - pending U.S. Provisional Patent Application No. 61 / 869,039, filed on August 22, 2013, and U.S. Provisional Patent Application No. 61 / 870,747, filed on August 27, 2013, each of which is incorporated herein by reference. Background

[0002] Multiple myeloma (MM), sometimes referred to as plasmacytic myeloma, is a multiple plasma cell cancer of the bone system and generally occurs in elderly individuals. When diagnosed, most individuals are symptomatic. Diagnosis is typically performed by one or more of serum protein electrophoresis, serum - free κ / λ light chain assay, urine protein electrophoresis (99% of MM patients show an increase in the level of one of the immunoglobulin (Ig) classes in the blood and / or light chains in the urine), bone marrow examination, or X - ray analysis. MM generally responds to chemotherapy, but recurrence is common because such treatment does not target cancer stem cells.

[0003] Nara et al. have recently identified several candidate genes that target the tumor - initiating sub - population (SP) cells, i.e., cancer stem cells, of MM. These include several genes that encode proteins related to the cell cycle and mitosis, all of which have been found to be up - regulated in MM cells. These include cyclin B1 (CCNB1), cell division cycle 2 (CDC2), baculoviral IAP repeat - containing 5 (BIRC5), abnormal spindle homolog, microcephaly - associated (ASPM), topoisomerase (DNA) II alpha 170 kDa (TOP2A), aurora kinase B (AURKB), kinesin family member 11 (KIF11), and kinesin family member 2c (KIF2C).

[0004] Similarly, Shaughnessy et al. reported a 70-gene high-risk profile for multiple myeloma. Two of the genes upregulated in this high-risk profile are CDC28 protein kinase regulatory subunit 1B (CKS1B) and WEE1 homolog (S. pombe) (WEE1). Overview

[0005] One embodiment of the present invention provides a method for treating multiple myeloma (MM) in an individual, the method comprising administering to the individual an effective amount of trichostatin A (TSA).

[0006] In another embodiment, the present invention provides a method for treating multiple myeloma (MM) in an individual, the method comprising determining the level of expression of at least one gene selected from the group consisting of CCNB1, AURKB, CDC2, BIRC5, KIF11, KIF2C, TOP2A, ASPM, CKS1B, and WEE1 from a biological sample obtained from the individual's body, and administering to the individual an effective amount of trichostatin A (TSA) when the level of expression of the at least one gene exhibits overexpression.

[0007] In yet another embodiment, the present invention provides a method for treating multiple myeloma (MM) in an individual, the method comprising diagnosing or already having diagnosed the individual with MM, and administering to the individual an effective amount of trichostatin A (TSA).

[0008] In yet another embodiment, the present invention provides a pharmaceutical composition comprising trichostatin A (TSA) as an inhibitor of CCNB1, AURKB, CDC2, BIRC5, KIF11, KIF2C, TOP2A, ASPM, CKS1B, or WEE1 alone or predominantly, and a pharmaceutically acceptable excipient or carrier.

[0009] In yet another embodiment of the present invention, the treatment with TSA is combined with one or more other multiple myeloma treatments. Such other treatments may include, for example, small molecule inhibition. Detailed description

[0010] Trichostatin A (TSA or 7-[4-(dimethylamino)phenyl]-N-hydroxy-4,6-dimethyl-7-oxohepta-2,4-dienamide) is an antifungal antibiotic. The structure of TSA is shown in the following formula I.

Chemical formula

[0011] The applicant has surprisingly found that although TSA has hitherto been known as a class I and II histone deacetylase (HDAC) inhibitor, it can also inhibit the expression of CCNB1, AURKB, CDC2, BIRC5, KIF11, KIF2C, TOP2A, ASPM, CKS1B, and WEE1. Therefore, TSA can be used as a primary or sole inhibitor of one or more such genes in the treatment of MM.

[0012] Human retinal pigment epithelial cell lines were treated with trichostatin or vehicle for 24 hours, and gene expression for 22,238 probe sets covering 12,490 genes was generated using equipment from Affymetrix. The effect of trichostatin A on the expression of CCNB1, AURKB, CDC2, BIRC5, KIF11, KIF2C, TOP2A, ASPM, CKS1B, and WEE1 is shown in Table 1 below, indicating significant downregulation of the expression of each gene.

[0013] Table 1:

Table 1

[0014] These results support the use of TSA in the treatment of MM. For example, an individual's MM can be treated by administering to the individual an effective amount of TSA, where an effective amount is an amount sufficient to inhibit the expression of one or more of CCNB1, AURKB, CDC2, BIRC5, KIF11, KIF2C, TOP2A, ASPM, CKS1B, and WEE1 in the individual. Such an amount can also be sufficient to inhibit HDAC activity in the individual. In some embodiments of the invention, the effective amount is from about 0.01 mg / kg / day to about 100 mg / kg / day, such as from about 0.1 mg / kg / day to about 10 mg / kg / day, or from about 0.5 mg / kg / day to about 5 mg / kg / day.

[0015] In some embodiments, treating the individual can further comprise determining the level of expression of one or more of CCNB1, AURKB, CDC2, BIRC5, KIF11, KIF2C, TOP2A, ASPM, CKS1B, or WEE1 from a biological sample obtained from the individual's body. Such determination can include any known or later-developed method or technique, such as quantitative antigen-antibody interaction, use of labeled nucleotide probes, and the like.

[0016] In other embodiments of the invention, treating the individual can include diagnosing the individual with MM or already having diagnosed the individual with MM prior to administering TSA to the individual. Such diagnosis can include one or more techniques or methods for making such a diagnosis, including, for example, serum protein electrophoresis, serum-free κ / λ light chain assay, urine protein electrophoresis, bone marrow examination, or X-ray analysis.

[0017] TSA can be administered to the subject to be treated in the form of a pharmaceutical composition. The pharmaceutical compositions used according to various embodiments of the present invention comprise a therapeutically effective amount of TSA or an active metabolite of TSA or a pharmaceutically acceptable salt or other form thereof (e.g., solvate) together with one or more pharmaceutically acceptable excipients or carriers. The term "pharmaceutical composition" refers to a composition suitable for administration in medical use. It should be understood that the determination of the appropriate dosage form, dosage, and route of administration for a particular patient is within the scope of the ordinary skill level in the technical fields of pharmacy and medicine.

[0018] Administration can be oral, but other routes of administration, such as parenteral, nasal, buccal, transdermal, sublingual, intramuscular, intravenous, rectal, vaginal, etc. can also be employed. Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In such solid dosage forms, the compound is mixed with at least one inert pharmaceutically acceptable excipient. Such excipients are, for example, (a) fillers or bulking agents, such as starch, lactose, sucrose, glucose, mannitol, and silicic acid, (b) binders, such as carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose, and acacia, (c) humectants, such as glycerol, (d) disintegrants, such as agar, calcium carbonate, potato or tapioca starch, alginic acid, some complex silicates, and sodium carbonate, (e) dissolution retardants, such as paraffin, (f) absorption promoters, such as quaternary ammonium compounds, (g) wetting agents, such as cetyl alcohol and glycerol monostearate, (h) adsorbents, such as kaolin and bentonite, and (i) lubricants, such as talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium lauryl sulfate, or mixtures thereof. In the case of capsules, tablets, and pills, the dosage form may contain buffering agents. Solid dosage forms such as tablets, coated tablets, capsules, pills, and granules can also be prepared using coatings and shells (e.g., enteric coatings and others well known in the art). The solid dosage forms may contain opacifying agents and may also be compositions that release one or more active compounds in a delayed manner in a specific part of the intestinal tract. Examples of embeddable compositions that can be used are polymeric substances and waxes. The active compound can also be in microencapsulated form, optionally containing one or more of the above excipients. Such solid dosage forms generally may contain from 1% to 95% (weight / weight) of the active compound. In certain embodiments, the active compound is in the range of 5% to 70% (weight / weight).

[0019] Solid compositions for oral administration can be formulated into unit dosage forms containing from about 0.1 mg to about 5000 mg of the active ingredient per dosage. The term "unit dosage form" refers to physically discrete units suitable as unit doses for human subjects and other mammals, where each unit contains a predetermined quantity of the active ingredient calculated to produce the desired effect over a period of treatment, together with the required pharmaceutical carrier. The TSA can be formulated into unit dosage forms, for example, capsules having from 0.1 to 5000 mg of the active ingredient in addition to excipients.

[0020] Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups, and elixirs. In addition to the compound or composition, the liquid dosage form may contain inert diluents commonly used in the art. Such inert diluents are, for example, water or other solvents, solubilizing and emulsifying agents such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3 - butylene glycol, dimethylformamide, oils (especially cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil, and sesame oil), glycerol, tetrahydrofurfuryl alcohol, polyethylene glycol, and fatty acid esters of sorbitan, or mixtures of these substances. In addition to such inert diluents, the composition may also contain adjuvants such as wetting agents, emulsifying agents, suspending agents, sweetening agents, flavoring agents, and fragrances.

[0021] In some embodiments of the present invention, the TSA is provided in liquid form and administered intravenously to an individual. According to some embodiments of the present invention, the TSA is provided in the form of a sustained or controlled release formulation.

[0022] The present invention has been described in conjunction with specific embodiments outlined above, but many alternative, modified, and variant forms will be apparent to those skilled in the art or are otherwise intended to be encompassed. That is, the embodiments of the present invention described above are for illustrative purposes and not for purposes of limitation. Various changes can be made without departing from the spirit and scope of the present invention as defined in the claims. All patents, patent applications, scientific papers, and other published documents cited herein are hereby incorporated by reference in their entirety for the content of their disclosure. Embodiments of the present invention include, for example, the following Embodiments 1 to 42. Embodiment 1: A method for treating multiple myeloma (MM) in an individual, comprising administering to the individual an effective amount of trichostatin A (TSA). A method comprising. Embodiment 2: The effective amount is an amount sufficient to reduce the expression of at least one gene in the individual, and the at least one gene is selected from the group consisting of CCNB1, AURKB, CDC2, BIRC5, KIF11, KIF2C, TOP2A, ASPM, CKS1B, and WEE1, according to the method of Embodiment 1. Embodiment 3: The effective amount is an amount sufficient to reduce the expression of either or both of CCNB1 and AURKB in the individual, according to the method of Embodiment 2. Embodiment 4: The effective amount is an amount sufficient to reduce the expression of either or both of CKS1B and WEE1 in the individual, according to the method of Embodiment 2. Embodiment 5: The effective amount is an amount sufficient to reduce the expression of each of CCNB1, AURKB, CDC2, BIRC5, KIF11, KIF2C, TOP2A, ASPM, CKS1B, and WEE1 in the individual, according to the method of Embodiment 2. Embodiment 6: The effective amount is about 0.01 mg / kg / day to about 100 mg / kg / day, according to the method of Embodiment 1. Embodiment 7: The method according to Embodiment 1, wherein the effective amount is from about 0.1 mg / kg / day to about 10 mg / kg / day. Embodiment 8: The method according to Embodiment 7, wherein the effective amount is from about 0.5 mg / kg / day to about 5 mg / kg / day. Embodiment 9: The method according to Embodiment 1, wherein the administration includes oral administration. Embodiment 10: The method according to Embodiment 1, wherein the administration includes intravenous administration. Embodiment 11: A method for treating multiple myeloma (MM) in an individual, comprising: determining the level of expression of one or more genes in an individual from a biological sample obtained from the individual's body, wherein the one or more genes are selected from the group consisting of CCNB1, AURKB, CDC2, BIRC5, KIF11, KIF2C, TOP2A, ASPM, CKS1B, and WEE1; administering an effective amount of trichostatin A (TSA) to the individual when the level of expression of the one or more genes exhibits overexpression; and a method comprising the above steps. Embodiment 12: The method according to Embodiment 11, wherein the effective amount is an amount sufficient to reduce the expression of at least one gene in the individual, and the at least one gene is selected from the group consisting of CCNB1, AURKB, CDC2, BIRC5, KIF11, KIF2C, TOP2A, ASPM, CKS1B, and WEE1. Embodiment 13: The method according to Embodiment 12, wherein the effective amount is an amount sufficient to reduce the expression of either or both of CCNB1 and AURKB in the individual. Embodiment 14: The method according to Embodiment 12, wherein the effective amount is an amount sufficient to reduce the expression of either or both of CKS1B and WEE1. Embodiment 15: The method according to embodiment 12, wherein the effective amount is an amount sufficient to reduce the expression of each of CCNB1, AURKB, CDC2, BIRC5, KIF11, KIF2C, TOP2A, ASPM, CKS1B, and WEE1. Embodiment 16: The method according to embodiment 11, wherein the effective amount is from about 0.01 mg / kg / day to about 100 mg / kg / day. Embodiment 17: The method according to embodiment 16, wherein the effective amount is from about 0.1 mg / kg / day to about 10 mg / kg / day. Embodiment 18: The method according to embodiment 17, wherein the effective amount is from about 0.5 mg / kg / day to about 5 mg / kg / day. Embodiment 19: The method according to embodiment 11, wherein the administration includes oral administration. Embodiment 20: The method according to embodiment 11, wherein the administration includes intravenous administration. Embodiment 21: A method for treating multiple myeloma (MM) in an individual, comprising: diagnosing or having already diagnosed the individual with MM; and administering to the individual an effective amount of trichostatin A (TSA). A method comprising the steps of: Embodiment 22: Diagnosing or having already diagnosed the individual includes performing or having already performed at least one diagnostic test on the individual, and the at least one diagnostic test is selected from the group consisting of serum protein electrophoresis, serum-free κ / λ light chain assay, urine protein electrophoresis, bone marrow examination, and X-ray analysis. The method according to embodiment 21. Embodiment 23: The method according to embodiment 21, wherein the effective amount is an amount sufficient to reduce the expression of at least one gene in the individual, and the at least one gene is selected from the group consisting of CCNB1, AURKB, CDC2, BIRC5, KIF11, KIF2C, TOP2A, ASPM, CKS1B, and WEE1. Embodiment 24: The method according to embodiment 23, wherein the effective amount is an amount sufficient to reduce the expression of either or both of CCNB1 and AURKB in an individual. Embodiment 25: The method according to embodiment 23, wherein the effective amount is an amount sufficient to reduce the expression of either or both of CKS1B and WEE1. Embodiment 26: The method according to embodiment 23, wherein the effective amount is an amount sufficient to reduce the expression of each of CCNB1, AURKB, CDC2, BIRC5, KIF11, KIF2C, TOP2A, ASPM, CKS1B, and WEE1. Embodiment 27: The method according to embodiment 21, wherein the effective amount is from about 0.01 mg / kg / day to about 100 mg / kg / day. Embodiment 28: The method according to embodiment 27, wherein the effective amount is from about 0.1 mg / kg / day to about 10 mg / kg / day. Embodiment 29: The method according to embodiment 28, wherein the effective amount is from about 0.5 mg / kg / day to about 5 mg / kg / day. Embodiment 30: The method according to embodiment 21, wherein the administration includes oral administration. Embodiment 31: The method according to embodiment 21, wherein the administration includes intravenous administration. Embodiment 32: Trichostatin A (TSA) as an inhibitor of at least one of CCNB1, AURKB, CDC2, BIRC5, KIF11, KIF2C, TOP2A, ASPM, CKS1B, or WEE1, a pharmaceutically acceptable excipient or carrier, and a pharmaceutical composition comprising the same. Embodiment 33: The pharmaceutical composition according to embodiment 32, wherein the effective amount is an amount sufficient to reduce the expression of either or both of CCNB1 and AURKB in an individual. Embodiment 34: The pharmaceutical composition according to embodiment 32, wherein the effective amount is an amount sufficient to reduce the expression of either or both of CKS1B and WEE1. Embodiment 35: The pharmaceutical composition according to embodiment 32, wherein TSA is an inhibitor of CCNB1, AURKB, CDC2, BIRC5, KIF11, KIF2C, TOP2A, ASPM, CKS1B, and WEE1 alone or mainly. Embodiment 36: The pharmaceutical composition according to embodiment 32, which is formulated for oral administration. Embodiment 37: The pharmaceutical composition according to embodiment 32, which is formulated for intravenous administration. Embodiment 38: The pharmaceutical composition according to embodiment 32, which contains TSA in an amount of about 0.1 mg to about 5000 mg. Embodiment 39: The pharmaceutical composition according to embodiment 32, wherein the amount of TSA corresponds to a dosage of about 0.01 mg / kg / day to about 100 mg / kg / day. Embodiment 40: The pharmaceutical composition according to embodiment 39, wherein the amount of TSA corresponds to a dosage of about 0.1 mg / kg / day to about 10 mg / kg / day. Embodiment 41: The pharmaceutical composition according to embodiment 40, wherein the amount of TSA corresponds to a dosage of about 0.5 mg / kg / day to about 5 mg / kg / day. Embodiment 42: The pharmaceutical composition according to embodiment 32, wherein the amount of TSA is effective to inhibit histone deacetylase (HDAC) activity.

Claims

**Claim 1** An agent for reducing the expression level of at least one gene determined to be overexpressed in an individual, the agent containing trichostatin A (TSA) as an active ingredient and being used such that an effective amount of TSA is administered to the individual, the at least one gene being selected from the group consisting of AURKB, CDC2, BIRC5, KIF11, KIF2C, TOP2A, ASPM, CKS1B, and WEE1. **Claim 2** The agent according to claim 1, wherein the effective amount is 0.1 mg / kg / day to 10 mg / kg / day. **Claim 3** The agent according to claim 2, wherein the effective amount is 0.5 mg / kg / day to 5 mg / kg / day. **Claim 4** The agent according to any one of claims 1 to 3, wherein the at least one gene is AURKB. **Claim 5** The agent according to any one of claims 1 to 3, wherein the at least one gene is CKS1B. **Claim 6** The agent according to any one of claims 1 to 3, wherein the at least one gene is WEE1. **Claim 7** The at least one gene is (a) AURKB, or (b) either or both of CKS1B and WEE1 The agent according to any one of claims 1 to 3.

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