Method of inhibiting the expression of a multi-drug resistance genes and inhibiting the production of proteins resulting from the expression of such genes thereby enhancing the effectiveness of chemot

Abstract

The present invention provides, in one aspect, a method of inhibiting the expression of a multi-drug resistance gene in an animal cell which comprises administering to an animal an effective amount of at least one cholesterol absorption inhibitor. In another aspect, it provides a method of inhibiting the production of a protein expressed by a multi-drug resistance gene in an animal cell which comprises administering to an animal an effective amount of at least one cholesterol absorption inhibitor. In another aspect, the present invention provides a method of enhancing the effectiveness of a chemotherapeutic agent in an animal having cancer, which comprises administering to said animal an effective amount of the chemotherapeutic agent and at least one cholesterol absorption inhibitor. Further, there are provided compositions and kits for use in cancer treatment which comprise at least one chemotherapeutic agent and at least one cholesterol absorption inhibitor.

Description

field of invention

[0001] The present invention relates to the field of cancer therapy. Background technique

[0002] It has now been found that the treatment of human malignancies with certain chemotherapeutic agents is often ineffective due to endogenous or acquired tumor cell resistance. The problem is exacerbated by multiple tumor cell resistance mechanisms leading to parallel resistance to a large number of structurally unrelated chemotherapeutic drugs with different mechanisms of action. This situation is known as multidrug resistance. This is an important problem in cancer treatment and appears to be a major obstacle to improving cure rates with current chemotherapeutic agents.

[0003] Approximately 2.04 million new cancer cases are reported annually in just seven major worldwide drug markets (US, Japan, Germany, Italy, France, Spain, UK) 1 . Unfortunately, only 5-10% of these respond successfully to chemotherapy. Approximately 40-45% or more than 800,000 patien...

Examples

Embodiment 1

[0225] Example 1 - A Cholesterol Absorption Inhibitor as described herein: Preparation of ascorbyl stanolyl phosphate referred to herein as "FM-VP4";

[0226] Step 1: Protect Ascorbic Acid

[0227] Fuming sulfuric acid (24%, 8.3g) was added dropwise to acetone (50ml). Ascorbic acid (12 g) was introduced into the mixture at 0°C, and the reaction mixture was stirred at 0°C for 6 hours. The resulting crystals were filtered with suction, and the filter cake was pressed dry and washed with acetone (30 ml). The product, 5,6-isopropylidene ascorbic acid (14 g) was obtained.

[0228] Step 2: Linking to Phytostanols

[0229] At 0°C, a solution of phytostanol mixture (24g) (camesteranol: 36.4%; sitostanol: 62.3%) in toluene (500ml) and pyridine (25ml) was added dropwise to phosphorus oxychloride ( 9ml) in a mixture in toluene (200mol). The mixture was stirred at room temperature for 3 hours. Pyridine hydrochloride was filtered off, and the mother liquor was concentrated to recover...

Embodiment 2

[0232] Example 2: - a kind of cholesterol absorption inhibitor described in the text: preparation of disodium ascorbyl phosphate of dehydroepiandrosterone

[0233] Acetone (150 ml) and L-ascorbic acid (50 g) were added to a dry round bottom flask at 0°C. Acetyl chloride (7.5ml) was added dropwise via addition funnel over 10 minutes. The reaction mixture was stirred at 0 °C for 24 hours. The precipitate was filtered off and washed with acetone (3 x 20ml). The white product, 5,6-isopropylidene ascorbic acid, was dried under vacuum for 1.5 hours to obtain a dry powder (52 g), yield 85%.

[0234] A dry three-neck round bottom flask was equipped with a stir bar, argon inlet and addition funnel. A solution of dehydroepiandrosterone (1.73 g, 6 mmol) in anhydrous THF (15 ml) and pyridine (2.4 ml) was added dropwise to anhydrous THF (12 ml) and POCl at 0 °C over 10 minutes 3 (0.7ml, 7.5mmol) in a mixture. A white precipitate formed immediately. The suspension was stirred for 40 m...

Embodiment 3

[0238] Example 3 - Another cholesterol absorption inhibitor described herein: Synthesis of disodium ascorbyl phosphate of 5α-androstane-3β-ol-17-one

[0239] To a dry round bottom flask was added 5α-androstane-3β-ol-17-one (1.0 g, 3.4 mmol), THF (8.6 ml) and pyridine (1.38 ml). The mixture was stirred at room temperature until a clear solution was obtained. Into another dry round bottom flask was added THF (6.9ml) and POCl 3 (0.4ml, 4.25mmol), stirred at 0°C for 5 minutes. To this mixture was added dropwise the 5α-androstane-3β-ol-17-one solution prepared above under an argon atmosphere over 10 minutes. After the addition was complete, the white suspension was stirred at 0°C for 35 minutes and at room temperature for 2 hours. The reaction was stopped and the white suspension was used for the coupling reaction without filtration.

[0240] 5,6-Isopropylidene ascorbic acid (2.0 g, 9.52 mmol) was dissolved in pyridine (1.71 ml) and THF (17 ml). The round bottom flask containi...

field of invention

[0001] The present invention relates to the field of cancer therapy. Background technique

[0002] It has now been found that the treatment of human malignancies with certain chemotherapeutic agents is often ineffective due to endogenous or acquired tumor cell resistance. The problem is exacerbated by multiple tumor cell resistance mechanisms leading to parallel resistance to a large number of structurally unrelated chemotherapeutic drugs with different mechanisms of action. This situation is known as multidrug resistance. This is an important problem in cancer treatment and appears to be a major obstacle to improving cure rates with current chemotherapeutic agents.

[0003] Approximately 2.04 million new cancer cases are reported annually in just seven major worldwide drug markets (US, Japan, Germany, Italy, France, Spain, UK) 1 . Unfortunately, only 5-10% of these respond successfully to chemotherapy. Approximately 40-45% or more than 800,000 patien...

Examples

Embodiment 1

[0225] Example 1 - A Cholesterol Absorption Inhibitor as described herein: Preparation of ascorbyl stanolyl phosphate referred to herein as "FM-VP4";

[0226] Step 1: Protect Ascorbic Acid

[0227] Fuming sulfuric acid (24%, 8.3g) was added dropwise to acetone (50ml). Ascorbic acid (12 g) was introduced into the mixture at 0°C, and the reaction mixture was stirred at 0°C for 6 hours. The resulting crystals were filtered with suction, and the filter cake was pressed dry and washed with acetone (30 ml). The product, 5,6-isopropylidene ascorbic acid (14 g) was obtained.

[0228] Step 2: Linking to Phytostanols

[0229] At 0°C, a solution of phytostanol mixture (24g) (camesteranol: 36.4%; sitostanol: 62.3%) in toluene (500ml) and pyridine (25ml) was added dropwise to phosphorus oxychloride ( 9ml) in a mixture in toluene (200mol). The mixture was stirred at room temperature for 3 hours. Pyridine hydrochloride was filtered off, and the mother liquor was concentrated to recover...

Embodiment 2

[0232] Example 2: - a kind of cholesterol absorption inhibitor described in the text: preparation of disodium ascorbyl phosphate of dehydroepiandrosterone

[0233] Acetone (150 ml) and L-ascorbic acid (50 g) were added to a dry round bottom flask at 0°C. Acetyl chloride (7.5ml) was added dropwise via addition funnel over 10 minutes. The reaction mixture was stirred at 0 °C for 24 hours. The precipitate was filtered off and washed with acetone (3 x 20ml). The white product, 5,6-isopropylidene ascorbic acid, was dried under vacuum for 1.5 hours to obtain a dry powder (52 g), yield 85%.

[0234] A dry three-neck round bottom flask was equipped with a stir bar, argon inlet and addition funnel. A solution of dehydroepiandrosterone (1.73 g, 6 mmol) in anhydrous THF (15 ml) and pyridine (2.4 ml) was added dropwise to anhydrous THF (12 ml) and POCl at 0 °C over 10 minutes 3 (0.7ml, 7.5mmol) in a mixture. A white precipitate formed immediately. The suspension was stirred for 40 m...

Embodiment 3

[0238] Example 3 - Another cholesterol absorption inhibitor described herein: Synthesis of disodium ascorbyl phosphate of 5α-androstane-3β-ol-17-one

[0239] To a dry round bottom flask was added 5α-androstane-3β-ol-17-one (1.0 g, 3.4 mmol), THF (8.6 ml) and pyridine (1.38 ml). The mixture was stirred at room temperature until a clear solution was obtained. Into another dry round bottom flask was added THF (6.9ml) and POCl 3 (0.4ml, 4.25mmol), stirred at 0°C for 5 minutes. To this mixture was added dropwise the 5α-androstane-3β-ol-17-one solution prepared above under an argon atmosphere over 10 minutes. After the addition was complete, the white suspension was stirred at 0°C for 35 minutes and at room temperature for 2 hours. The reaction was stopped and the white suspension was used for the coupling reaction without filtration.

[0240] 5,6-Isopropylidene ascorbic acid (2.0 g, 9.52 mmol) was dissolved in pyridine (1.71 ml) and THF (17 ml). The round bottom flask containi...

Images