WO2007014652A1 - TREATMENT OF TUMOURS WITH IKK-ß INHIBITORS - Google Patents

Abstract

The application relates to the use of an IKK-beta inhibitor through production of medicaments for treating tumours, and to medicaments comprising the said IKK-beta inhibitor.

Description

Treatment of tumors with IKK-ß inhibitors

The application relates to the use of an IKK-beta inhibitor by the preparation of medicaments for the treatment of tumors, as well as medicaments containing this DCK-beta inhibitor.

The treatment of tumors of the skin represents a large and, especially concerning the metastatic melanoma, unsatisfactorily solved medical problem.

The most common tumors of fair-skinned humans are epithelial skin tumors, e.g. Basal cell carcinomas and spinozllular carcinomas including their pre-invasive precursors (actinic keratoses or carcinoma in situ).

While basaliomas and spinaliomas together account for about 15 percent of all malignant tumors, melanoma occurs much less frequently. According to the Dachdokumentation Krebs in the Robert Koch Institute, almost 11,500 illnesses occurred in Germany in the year 2000, in the USA the estimated number of new cases in 2003 was about 54,000. The prognosis of metastatic melanoma is still unfavorable and There is usually little chance of permanent cure.

Among the major mechanisms responsible for the malignancy of metastatic skin tumors is resistance to apoptosis (programmed cell death).

Therefore, therapeutic strategies, or substances that can re-induce apoptosis in tumors, are of prime importance for successful therapies of tumors of the skin (1).

OBJECT OF THE INVENTION Investigations have thus been the development of a new therapeutic agent for tumorous skin diseases, which does not have the drawbacks of systemic chemotherapy (for example considerable side effects, short-term clinical improvement, often not simple application) and induces apoptosis in cancer cells.

It has now surprisingly been found that the compound of the formula (I)

and / or their salts, hydrates and solvates, which are potent, competitive and selective inhibitors of UCK-beta kinase, can be used to treat tumors and their precursors. Preferably, the hydrochloride of the compound of formula (I).

The compound of formula (I) and / or its salts, hydrates and solvates, thereby act apoptosis-inducing in the melanoma cell line A375, which is sensitive to various pro-apoptotic stimuli. If in cells which are sensitive to apoptosis in response to pro-apoptotic stimuli, caspases (enzymes which mediate intracellular apoptosis pathways) are blocked with the inhibitor zVAD, the apoptosis induced in these cells by stimulation with CH-I l is suppressed. In contrast, the addition of the compound of formula (I) and / or its salts, hydrates and solvates may re-induce apoptosis, indicating alternative routes of induction of apoptosis.

In particular, it has been found that the compound of formula (I) and / or its salts, hydrates and solvates are useful for the treatment of skin tumors, e.g. Basaliomas, spinaliomas, melanomas, cutaneous manifestations of T-cell lymphomas, cutaneous metastases of other tumors and their precursors, in particular actinic keratoses and Bowen's disease, can be used.

The treatment is preferably systemic and / or topical. In a systemic treatment, the application of the drug can be made orally with tablets, juices, emulsions, capsules or other pharmaceutical preparations. Systemic treatment may also be parenteral (intravenous, subcutaneous). The topical treatment is carried out by applying the active ingredient in suitable formulations on the affected skin (skin lesion) or in the vicinity of the skin lesions to be treated. The formulations used may be ointments, creams, powders, emulsions, solutions. In addition, drug-impregnated patches or dressings may be used. The systemic and / or topical treatment can be carried out according to different time schedules (one-time treatment up to several times a day over a defined period of time).

Preference is given to the topical treatment of tumors of the eye and metastases of other tumors on the eye as well as of tumors of the ear (and metastases of other tumors on the ear) and of Tumors of the external genitalia (and metastases of other tumors on the external genitalia). This includes tumors caused by viral infections (eg genital warts caused by infection with different types of human papilloma virus, skin and mucous capillary cavities, epidermodysplasia verucciformis or other neoplastic changes ) one.

A further subject of the application are combinations of the compound of the formula (I) and / or its salts, hydrates and solvates, with at least one other active substance which can be used for the treatment of tumor diseases, in particular with dacarbacin (DTIC-DOME), doxorubicin ( Doxil), interferons (eg, interferon alfa-2b, intron A), imiquimod (Aldara, R-837, S-26308), resiquimod (R-848, S-28436), interleukin 2 (IL-2, proleukin, aldesleukin ), Retinoids, temozolomide and / or therapeutic melanoma vaccines (eg, prepared or synthetic antigens, or transfected cells) for the treatment of skin tumors.

A combined treatment of the compound of formula (I) and / or its salts, hydrates and ^"" solvates, with ^" radiotherapies; Irradiation therapy with visible light, ionizing radiation, UV radiation, photodynamic therapy is also preferred.

Topical treatment of tumors of the genitourinary system e.g. by instillation of a solution or suspension of the compound of formula (I) or other suitable pharmaceutical preparations of the compound of formula (I) (e.g., emulsions, gels) is also possible.

The following examples serve to illustrate the invention.

1. IKK Beta Kinase Assay:

1. IKK beta kinase assay:

The kinase activity of human IKK-beta (7 nM) is measured at different concentrations [Y ³³ P] ATP and GST-IκBα (1-54) substrate in the presence of the compound of formula (I) = substance A. Fixed concentrations of 2 μM GST-IκBα (1-54) (Figure 3 A) and 5 μM ATP (Figure 3 B) are chosen. It is incubated for 30 minutes at 25 ⁰ C, the reaction is then stopped by addition of EDTA. With TCA, the reaction is precipitated in a filter plate, Microscint PS (Packard) is added and the amount of ³³ P incorporated in GST-IκBα (1-54) is determined with a TopCount (Packard).

2. Measurement of apoptosis activity in melanoma cell lines

A375 cells are plated in a density of 3x10 ⁶ cells and Mel2a cells at a density of 3x10 ³ cells in triplets in cell culture dishes and over the period indicated in Figures 3 to 5 with the compound of formula (I) or the corresponding controls, 0.5 μg / ml of anti-CD-95 (CH-I, Fa. Upstate), 6.6 μM zVAD (from R & D Systems). The addition takes place 30 minutes before the second substance. Thereafter, the detection of apoptosis with a Cell Death Detection ELISA ^PLUS (Roche Diagnostics) according to the manufacturer. A375 and Mel2a cells are established melanoma cell lines.

Apoptosis assay on melanoma cell line A375 after 24 h incubation

0.1% 57.88 28.94 13.89 5.78 1.38 0.138 0.00552 CH 11

DMSO

Bay 65-5811 Concentration in Incubation Medium [μM]

Figure 3: Detection of apoptosis in A375 melanoma cells. Substance A induces dose-dependent apoptosis. Positive control: The antibody CH-I l stimulates Fas (CD-95) and induces apoptosis.

Apoptosis assay on melanoma cell line Mel2a after 24h incubation

0.1% 0.2% 57.88 28.94 13.89 5.78 1.38 0.138 0.00552 CH 11

DMSO DMSO

Bay 65-5811 concentration in incubation medium [μM]

Figure 4: Mel-2a cells are resistant to Fas stimulation (CD-95). Substance A, on the other hand, induced dose-dependent apoptosis in these cells, indicating a receptor-independent mechanism.

Apoptosis assay on melanoma cell line A375 after 12 h incubation

DMSO 0.2% Z-VAD CHIl Z-VAO + CH11 Z-VAD + 28.94 28.94

Bay 65-5811 concentration in incubate> πsmeflium [μM]

Figure 5: Inhibition of caspases with the inhibitor zVAD in apoptosis-sensitive (CH-I l) cells. The addition of A triggers the apoptosis inhibited by zVAD.

Claims

claims 1. Use of the compound of the formula (I)

and / or their salts, hydrates and solvates, for the preparation of medicaments for the treatment of tumors and their precursors. 2. Use according to claim 1, for the treatment of skin tumors and their precursors. 3. Use according to claim 1, for the preparation of systemic medicaments. 4. Use according to claim 1, for the preparation of topical medicaments. 5. Use according to claim 1, for the preparation of medicaments for the topical treatment of tumors of the eye and metastases of other tumors on the eye, of tumors of the ear and metastases of other tumors on the ear, of tumors of the outer genitalia and metastases of other tumors on the outer Genitals. 6. Medicaments for the treatment of tumors and their precursors containing a compound of the formula (I)

and / or their salts, hydrates and solvates. 7. Medicaments for the treatment of tumors of the skin and their precursors containing a Compound of the formula (I)

and / or their salts, hydrates and solvates. 8. Medicaments containing a compound of formula (I) and / or their salts, hydrates and solvates, in combination with at least one other active substance which can be used for the treatment of tumor diseases.