JP2009502995A - Treatment of tumors with IKK-β inhibitors - Google Patents

Abstract

  The present application relates to the use of an IKK-beta inhibitor by the manufacture of a medicament for the treatment of tumors, and to a medicament comprising said IKK-beta inhibitor.

Description

  The present application relates to the use of an IKK-beta inhibitor for the manufacture of a medicament for the treatment of tumors and to a medicament comprising this IKK-beta inhibitor.

  The treatment of skin tumor diseases is a medical problem that has been very unresolved, especially with regard to metastatic melanoma.

  Here, the most frequent tumors among the fair-skinned people are epithelial skin tumors such as basal cell carcinoma and squamous cell carcinoma, for example, and their preinvasive early stages (photokeratinization) Disease or carcinoma in situ).

  Basal cell tumors and squamous cell carcinoma together account for about 15 percent of all malignant tumors, but melanoma occurs more rarely. According to the Robert-Koch Institute's comprehensive evidence of cancer, nearly 11500 illnesses occurred in Germany in 2000, and the estimated number of new diseases in the United States in 2003 was approximately 54,000. . The prognosis for metastatic melanoma is still disadvantageous and generally has little prospect of permanent healing.

  The most important mechanisms responsible for the malignancy of metastatic skin tumors include resistance to apoptosis (programmed cell death).

  Therefore, therapeutic strategies or substances that can re-induce apoptosis in tumor cells are crucial for the successful treatment of skin tumors (1).

  Accordingly, the object of the present invention is a novel therapeutically active substance in neoplastic skin diseases, and the disadvantages of systemic chemotherapy (eg widespread side effects, clinical improvement that lasts only for a short time, often inconvenient administration) It was the development of a substance that induces apoptosis in cancer cells.

の化合物、および／またはその塩、水和物および溶媒和物を、腫瘍およびそれらの早期段階の処置に使用できることが見出された。式（Ｉ）の化合物の塩酸塩が好ましい。 Surprisingly, formula (I) is a potent, competitive and selective inhibitor of IKK-beta kinase.

It has been found that these compounds, and / or their salts, hydrates and solvates can be used for the treatment of tumors and their early stages. Preference is given to the hydrochloride of the compound of formula (I).

  The compounds of formula (I) and / or their salts, hydrates and solvates have an apoptosis-inducing action in the melanoma cell line A375 that reacts sensitively to various pro-apoptotic stimuli. Inhibition of the caspase of A375 cells (enzyme that mediates the intracellular signal transduction pathway of apoptosis), which reacts by causing apoptosis sensitively to pro-apoptotic stimuli, is inhibited in these cells by inhibiting the inhibitor zVAD. Apoptosis induced by stimulation is suppressed. However, the addition of compounds of formula (I) and / or their salts, hydrates and solvates can again induce apoptosis, indicating an alternative pathway for inducing apoptosis.

  In particular, the compounds of formula (I) and / or their salts, hydrates and solvates are used in skin tumors such as basal cell carcinoma, squamous cell carcinoma, melanoma, T-cell lymphoma skin manifestations, other It has been shown that it can be used for the treatment of tumor skin metastases and their early stages, for example, especially actinic keratosis and Bowen's disease.

  Treatment is preferably performed systemically and / or locally. In the case of systemic treatment, the administration of the active substance can be carried out orally using tablets, juices, emulsions, capsules or other pharmaceutical preparations. Systemic treatment can also be performed parenterally (intravenous, subcutaneous). Topical treatment is performed by applying the active substance in a suitable formulation to the affected skin (skin lesion) or in the vicinity of the skin lesion to be treated. The formulation used can be an ointment, cream, powder, emulsion, solution. In addition, patches or bandages soaked with the active substance can be used. Systemic and / or local treatment can be performed according to various time schedules (from a single treatment to several treatments per day over a defined period of time).

  Local treatment of ocular and other tumor eye metastases, as well as ear tumors (and other tumor ear metastases), and external genital tumors (and other tumor external genital metastases) are preferred. This includes tumors resulting from viral infections (eg, genital warts, skin and mucosal Kaposi's sarcoma, warts-like epidermal growth disorders or other neoplastic changes resulting from infection with various types of human papillomavirus) Is included.

  The invention further relates to a compound of formula (I) and / or a salt, hydrate and solvate thereof for the treatment of skin tumors and at least one other activity which can be used for the treatment of tumors. In particular, dacarbacin (DTIC-DOME), doxorubicin (Doxil), interferons (eg, interferon alpha-2b, intron-A), imiquimod (Aldara, R-837, S-26308), resiquimod (Resiquimod) (R-848, S-28436), interleukin 2 (IL-2, proleukin, aldesleukin), retinoids, temozolomide and / or therapeutic melanoma vaccines (eg prepared or synthetic antigens) Or in combination with transfected cells).

  Preference is likewise given to treatments combining the compounds of the formula (I) and / or their salts, hydrates and solvates with radiation therapy, irradiation therapy with visible light, ionizing radiation, UV irradiation, photodynamic therapy.

  For example, topical treatment of urogenital tumors by instillation of a solution or suspension of a compound of formula (I) or other suitable pharmaceutical formulation (eg, emulsion, gel) of a compound of formula (I) may include: It is possible as well.

The following examples serve to illustrate the invention.
1. IKK beta-kinase assay:
The kinase activity of human IKK-beta (7 nM) is measured in various concentrations of [γ ³³ P] ATP and GST-IκBα (1-54) substrate in the presence of compound of formula (I) = substance A. Here, a fixed concentration of 2 μM GST-IκBα (1-54) (FIG. 3A) or 5 μM ATP (FIG. 3B) is selected. The mixture is incubated for 30 minutes at 25 ° C., after which the reaction is stopped by the addition of EDTA. The reaction was precipitated in the filter plate using TCA, Microscint PS (Packard) was added, and the amount of ³³ P incorporated into GST-IκBα (1-54) was determined using TopCount (Packard). decide.

2. Measurement of Apoptotic Activity in Melanoma Cell Line A375 cells are seeded at a density of 3 × 10 ⁶ cells and Mel2a cells are seeded at a density of ³ × 10 ³ cells in a cell culture dish in a triplate and the compound of formula (I) or corresponding Incubate with 0.5 μg / ml anti-CD-95 (CH-11, Upstate), 6.6 μMzVAD (R & D Systems) for the period shown in FIGS. The addition was performed 30 minutes before the second material. Following this, apoptosis detection is performed using Cell Death Detection ELISA ^PLUS (Roche Diagnostics) according to the manufacturer's instructions. A375 and Mel2a cells are established melanoma cell lines.

図３：Ａ３７５黒色腫細胞におけるアポトーシスの検出。物質Ａは、用量依存的アポトーシスを誘導する。正の対照：抗体ＣＨ−１１は、Ｆａｓ（ＣＤ−９５）を刺激し、かくしてアポトーシスを誘導する。

FIG. 3: Detection of apoptosis in A375 melanoma cells. Substance A induces dose-dependent apoptosis. Positive control: Antibody CH-11 stimulates Fas (CD-95) and thus induces apoptosis.

図４：Ｍｅｌ−２ａ細胞は、Ｆａｓ（ＣＤ−９５）による刺激に対して耐性である。しかしながら、物質Ａは、これらの細胞で用量依存的アポトーシスを誘導した。これは、受容体非依存的メカニズムを示している。

FIG. 4: Mel-2a cells are resistant to stimulation by Fas (CD-95). However, substance A induced dose-dependent apoptosis in these cells. This indicates a receptor-independent mechanism.

図５：アポトーシス−感受性（ＣＨ−１１）細胞における阻害剤ｚＶＡＤによるカスパーゼの阻害。Ａの添加は、ｚＶＡＤにより阻害されるアポトーシスを再び誘導する。

FIG. 5: Caspase inhibition by the inhibitor zVAD in apoptosis-sensitive (CH-11) cells. Addition of A again induces apoptosis that is inhibited by zVAD.

Claims (8)

Formula (I)

And / or salts, hydrates and solvates thereof for the manufacture of a medicament for the treatment of tumors and their early stages.   Use according to claim 1 for the treatment of skin tumors and their early stages.   Use according to claim 1 for the manufacture of a systemic medicament.   Use according to claim 1 for the manufacture of a topical medicament.   Claims for the manufacture of a medicament for the local treatment of ocular tumors and ocular metastases of other tumors, ear tumors and ear metastases of other tumors, and external genital tumors and external tumor metastases of other tumors Item 1. Use according to Item 1. Formula (I)

And / or a salt, hydrate and solvate thereof for treating tumors and early stages thereof. Formula (I)

And / or a salt, hydrate and solvate thereof for treating tumors of the skin and early stages thereof.   A medicament comprising a compound of formula (I) and / or a salt, hydrate and solvate thereof in combination with at least one other active substance which can be used for the treatment of tumors.