A pharmaceutical compound, method of preparation thereof, and its use for treating cancer

Abstract

There is provides a compound of the formula (I) or a salt thereof, a method of preparation thereof, a pharmaceutical composition including the same, and its use for treating cancer.

Description

[0001] A PHARMACEUTICAL COMPOUND, METHOD OF PREPARATION THEREOF, AND ITS USE FOR TREATING CANCER TECHNICAL FIELD

[0001] The present disclosure relates to novel compounds, methods of preparation, pharmaceutical composition, and more particularly to novel compounds prepared by chemical conjugation process of two compounds, and their use for treating cancer. BACKGROUND INFORMATION

[0002] Doxorubicin is a chemotherapeutic agent that is frequently used in treatment of several cancers including breast, lung, gastric, ovarian, thyroid and lymphoma. Despite the wide use of doxorubicin in cancer treatment, the drug has demonstrated severe side effects such as cardiotoxicity, bone marrow aplasia and nephrotoxicity.

[0003] On the other hand, thymoquinone is a herbal-derived drug with known anti-oxidant, chemopreventive and chemotherapeutic activities. However, thymoquinone suffers from high hydrophobicity causing poor solubility in aqueous medium which limits its bioavailability.

[0004] A study by Ozturk et al. titled “Thymoquinone is a protective agent that reduces the negative effects of doxorubicin in rat testis” reported that co-treatment of thymoquinone and doxorubicin could ameliorate doxorubicin-induced hepatoxicity and testicular toxicity in rats.

[0005] Other attempts in the art were made to develop pharmaceutical compositions containing thymoquinone and doxorubicin. For instance, the international application published under number WO2022130225 discloses a composition for use in treating cancer, the composition may include doxorubicin and thymoquinone.

[0006] Yet other attempts in the art were made to hybridize thymoquinone with other compounds. For instance, a study by Fröhlich et al. titled “Synthesis of Thymoquinone-Artemisinin Hybrids: New Potent Antileukemia, Antiviral and Antimalarial Agents” disclosed the chemical conjugation of thymoquinone with Artemisinin to provide antileukemia, antiviral and antimalarial agents.

[0007] The review study by Shah et al. titled “Structural modification and strategies for the enhanced doxorubicin drug delivery” discloses a review of the attempts reported in the art to modify doxorubicin in order to reduce its side effects. SUMMARY

[0008] It is an object of the present disclosure provide a novel hybridized compound of thymoquinone and doxorubicin having a structure according to the formula (I), or a salt thereof: Formula (I)

[0009] Other aspects of the present disclosure provide a method for preparing of the compound of the formula (I), wherein the method may include the steps of: - Dissolving thymoquinone and succinic acid in a mixture of acetonitrile and water under an inert atmosphere to provide a first mixture; - Adding silver nitrate to the first mixture under the inert atmosphere at room temperature to provide a second mixture; - Heating the second mixture under reflux, followed by a gradual addition of ammonium persulfate dissolved in distilled water to provide a third mixture; - Diluting the third mixture with distilled water, followed by extracting the diluted third mixture using diethylether as an extraction solvent; - Taking an organic layer resulting from the extraction step, drying it over anhydrous sodium sulfate powder, and evaporating such layer to produce thymoquinone-succinic acid conjugate; - Dissolving thymoquinone-succinic acid conjugate in dimethyl sulfoxide (“DMSO”) while stirring under nitrogen gas to provide a first solution; - Adding 1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide (“EDC”) to the first solution, followed by an addition of sulfo-NHS while stirring to provide a second solution; and - Adding doxorubicin-HCl dissolved in DMSO with a trace amount of trimethylamine to the second solution with magnetic stirring in a dark place under nitrogen to produce the compound of the formula (I).

[0010] Other aspects of the present disclosure provide a pharmaceutical composition including the compound of formula (I) and / or pharmaceutically acceptable salts thereof and a pharmaceutically acceptable carrier / excipient.

[0011] In aspects of the present disclosure, the pharmaceutical composition may be formulated as a solid, liquid, or semi-solid dosage form.

[0012] In some aspects, the pharmaceutical composition may be administered via different routes such as oral, parenteral, intramuscular, intranasal, sublingual, intratracheal, ocular, vaginal, rectal, or intraventricular.

[0013] In aspects of the present disclosure, the pharmaceutical composition may be used for treating cancer.

[0014] In aspects of the present disclosure, the pharmaceutical composition has high anti-tumor activity and low side toxicity on healthy cells. BRIEF DESCRIPTION OF THE DRAWINGS

[0015] The disclosure will now be described with reference to the accompanying drawings, which illustrate embodiments of the present disclosure, without however restricting the scope of the disclosure thereto, and in which:

[0016] FIG.1 illustrates a flowchart of a method of preparing a compound of formula (I), the method being configured in accordance with embodiments of the present disclosure.

[0017] FIG.2A illustrates a chart showing the1H-NMR spectrum for thymoquinone-succinic acid conjugate in d6-DMSO at a temperature of about 25oC, wherein such spectrum is obtained using Bruker Biospin AG Magnet system 500MHz / 54mm instrument.

[0018] FIG.2B illustrates a chart showing the13C-NMR spectrum for thymoquinone-succinic acid conjugate in d6-DMSO at a temperature of about 25oC, wherein such spectrum is obtained using Bruker Biospin AG Magnet system 500MHz / 54mm instrument.

[0019] FIG.2C illustrates a chart showing the13C-dept-135 NMR spectrum for thymoquinone- succinic acid conjugate in d6-DMSO at a temperature of about 25oC, wherein such spectrum is obtained using Bruker Biospin AG Magnet system 500MHz / 54mm instrument.

[0020] FIG.3A illustrates a high resolution mass spectrum (“HR-MS”) of thymoquinone- succinic acid conjugate on ([M+H+]+) using electrospray ion trap (“ESI”) technique by collision-induced dissociation on a Bruker APEX-4 (7 Tesla) instrument.

[0021] FIG.3B illustrates an HR-MS of thymoquinone-succinic acid conjugate on ([M+Na+]+) ESI technique by collision-induced dissociation on a Bruker APEX-4 (7 Tesla) instrument.

[0022] FIG.4A illustrates a chart showing the1H-NMR spectrum for the compound of the formula (I) in d6-DMSO at a temperature of about 25oC, wherein such spectrum is obtained using Bruker Biospin AG Magnet system 500MHz / 54mm instrument.

[0023] FIG.4B illustrates a chart showing the13C-dept- 135 NMR spectrum for the compound of the formula (I) in d6-DMSO at a temperature of about 25oC, wherein such spectrum is obtained using Bruker Biospin AG Magnet system 500MHz / 54mm instrument.

[0024] FIG.5 illustrates a chart showing the1H-15N-HMBC NMR spectrum for the compound of the formula (I) in CD3OD.

[0025] FIG.6A illustrates a mass spectrum of the compound of the formula (I) on ([M+H+]+).

[0026] FIG.6B illustrates a mass spectrum of the compound of the formula (I) on ([M+NA+]+).

[0027] FIG.7 illustrates high performance liquid chromatography (“HPLC”) analysis of the compound of the formula (I).

[0028] FIG.8 illustrates a line chart showing cytotoxicity of the compound of the formula (I), thymoquinone and doxorubicin on cancer cells, wherein (“TQ”) refers to thymoquinone, (“TQ-Dox”) refers to the compound of the formula (I), and (“Dox”) refers to doxorubicin.

[0029] FIG.9 illustrates line charts showing cytotoxicity of the compound of the formula (I), thymoquinone and doxorubicin on normal cells, wherein (“TQ”) refers to thymoquinone, (“TQ-Dox”) refers to the compound of the general formula (I), and (“Dox”) refers to doxorubicin.

[0030] FIG.10 shows the cellular uptake and quantification of subcellular localization of the compound of the formula (I) within estrogen receptors (“ER”) organelles, wherein (“TQ-Dox hybrid”) refers to the compound of the formula (I). DETAILED DESCRIPTION

[0031] Embodiments of the present disclosure provide a hybrid compound of thymoquinone and doxorubicin according to the formula (I), or a salt thereof.

[0002]

[0032] Reference now is being made to FIG.1, which illustrates a flowchart of a method for preparing the compound of the formula (I), wherein the method includes the steps of: - Dissolving thymoquinone and succinic acid in a mixture of acetonitrile and water under an inert atmosphere to provide a first mixture (process block 1-1); - Adding silver nitrate to the first mixture under the inert atmosphere at room temperature to provide a second mixture (process block 1-2); - Heating the second mixture under reflux, followed by a gradual addition of ammonium persulfate dissolved in distilled water to provide a third mixture (process block 1-3); - Diluting the third mixture with distilled water, followed by extracting the diluted third mixture using diethylether as an extraction solvent (process block 1-4); - Taking an organic layer resulting from the extraction step, drying it over anhydrous sodium sulfate powder, and evaporating such layer to produce thymoquinone-succinic acid conjugate (process block 1-5); - Dissolving thymoquinone-succinic acid conjugate in DMSO while stirring under nitrogen gas to provide a first solution (process block 1-6); - Adding 1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC) to the first solution, followed by an addition of sulfo-NHS while stirring to provide a second solution (process block 1-7); and - Adding doxorubicin-HCl dissolved in DMSO with a trace amount of trimethylamine to the second solution with magnetic stirring in a dark place under nitrogen to produce the compound of the formula (I) (process block 1-8).

[0033] Other embodiments of the present disclosure further provide a pharmaceutical composition including a compound of formula (I) and / or pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier / excipient.

[0034] The term “pharmaceutical composition”, as used herein, is intended to include a compound of formula (I) and / or a pharmaceutically acceptable salt thereof.

[0035] In embodiments of the present disclosure, the pharmaceutical composition can be, for example, in a liquid form, e.g. a solution, syrup, emulsion and suspension, or in a solid form, e.g. a capsule, caplet, tablet, pill, powder, and suppository. Granules, semi-solid forms, and gel caps are also considered. In case that the pharmaceutical composition is a liquid or a powder, dosage unit optionally is to be measured, e.g. in the dosage unit of a teaspoon.

[0036] The pharmaceutical composition in embodiments of the present disclosure can be formulated for oral administration in solid or liquid form, for parenteral injection or for rectal administration. The pharmaceutical composition can be administered to humans and other mammals orally, sublingually, rectally, parenterally, intracisternally, intraurethrally, intraperitoneally, topically (as powder, ointment or drop), as buccal or as an oral or nasal spray. The term "parenterally", as used herein, refers to modes of administration which include intravenous, intramuscular, intraperitoneal, subcutaneous, intra-articular injection and infusion.

[0037] The term “pharmaceutical acceptable carrier / excipient”, as used herein, means a non-toxic, inert solid, semi-solid or liquid filler, diluent, encapsulating material, or formulation auxiliary of any type. Some examples of materials which can serve as pharmaceutically acceptable carriers are sugars such as lactose, glucose and sucrose; starches such as corn starch; cellulose and its derivatives such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; powdered tragacanth; binding agents such as hypromellose; disintegrating agents such as crosscarmellose; water; malt; gelatin; talc; excipients such as cocoa butter and suppository waxes; oils such as peanut oil; cottonseed oil; safflower oil, sesame oil, olive oil, corn oil and soybean oil; glycols, such as propylene glycol; esters such as ethyl oleate and ethyl laurate; agar; buffering agents such as magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen-free water; isotonic saline; Ringer's solution; ethyl alcohol, and phosphate buffer solutions, as well as other non-toxic compatible lubricants such as sodium lauryl sulfate and magnesium stearate, as well as coloring agents, releasing agents, coating agents, sweetening, flavoring and perfuming agents, preservatives and antioxidants can also be present in the composition, according to the judgement of the formulator.

[0038] All components of the pharmaceutical composition have to be pharmaceutically acceptable. The term “pharmaceutically acceptable” means at least non-toxic.

[0039] According to embodiments of the present disclosure, the pharmaceutical composition may be used for treating cancer.

[0040] In some embodiments of the present disclosure, the pharmaceutical composition may be used for treating breast cancer.

[0041] The disclosure will be further illustrated on the basis of examples and a detailed description from which further features and advantages may be taken. It is to be noted that the following explanations are presented for the purpose of illustrating and description only; they are not intended to be exhaustive or to limit the disclosure to the precise form disclosed. Example 1 Chemical hybridization of thymoquinone with doxorubicin

[0042] The compound of the formula (I) can be prepared according to the following preparation scheme:

[0003]

[0043] About 150 mg of thymoquinone and about 87 mg succinic acid were added to a 25 ml two-neck flask. Then, the chemicals were dissolved in a mixture having about 9 ml of total volume of (1:1) acetonitrile (CH3CN): water, under argon gas. Directly, about 15 mg of silver nitrate (AgNO3) was added and the mixture was left about 2-3 minutes at room temperature under argon gas. Then, the reaction mixture was heated up to about 80oC with reflux for about 5 minutes. After that, about 208.5 mg of ammonium persulfate (NH4)2S2O8 dissolved in about 1.5 ml distilled water was added gradually. The reaction mixture was refluxed at 100oC / overnight. The reaction mixture was further diluted with about 0.9 ml distilled water. Then, diethyl ether was used as an extraction solvent in an extraction step. The organic layer was taken and dried over anhydrous sodium sulfate (Na2SO4) powder and evaporated at atmospheric pressure / overnight to produce thymoquinone-succinic acid conjugate. Then, about 30.0 mg, 0.1271 mmol of thymoquinone-succinic acid conjugate was dissolved in about 500 μL DMSO and stirred under nitrogen gas. To activate the carboxylic acid group, about 73.1 mg, 0.38 mmol of EDC was added to the thymoquinone- succinic acid conjugate solution followed by an addition of about 82.8 mg, 0.38 mmol Sulfo-NHS and stirred for about 3 hours. About 2.5 equivalents of doxorubicin-HCl (about 172.7 mg, 0.173 mmol) dissolved in about 500 μL DMSO with trace amounts of triethylamine (about 22-44 μL) were then added to reaction solution, with magnetic stirring for about 12-24 hours in dark place under nitrogen. Example 2 Characterization of the thymoquinone-succinic acid conjugate and the compound of the formula (I)

[0044] Reference in this example is being made to FIGS.2A-2C, 3A-3B, 4A-4B. Characterization of thymoquinone-succinic acid conjugate

[0045] FIGS.2A-2C illustrate 1H-NMR, 13C-NMR, and 13C-dept-135 NMR spectra for thymoquinone-succinic acid conjugate in d6-DMSO at a temperature of about 25oC. All NMR spectra were performed in d6-DMSO (99.0% atom, Across Organics, U.S.A.) using Bruker Biospin AG Magnet system 500MHz / 54mm instrument (Bruker BioSpin GmbH, Switzerland). Chemical shift in parts per million (“ppm”) were referenced to tetramethylsilane (“TMS”) as an internal reference. Data processing and analysis of the data were performed with topspin framework (Bruker Biospin GmbH, Version 3).

[0046] As illustrated in FIGS. 2A-2C, two additional CH2groups appeared at 2.21 and 2.64 ppm for protons, and 21.9 and 33.8 ppm for carbons. Also, the protons of the dimethyl appeared at 1.16 ppm as a doublet and the methyl at 1.91 ppm whereas the aromatic proton appeared at 6.55 ppm and the other signal almost disappeared at 6.49 ppm.

[0047] The formation of thymoquinone-succinic acid conjugate was also confirmed using high resolution mass spectrometry (“HRMS”) using electrospray ion trap (“ESI”) on the positive mode confirmed thymoquinone-succinic acid conjugate (C13H16O4) synthesis. HRMS were acquired using electrospray ion trap ESI technique by collision-induced dissociation on a Bruker APEX-4 (7 Tesla) instrument. The samples were infused using a syringe pump at a flow rate of 2 μL min−1.

[0048] the calculated and the found values of HRMS were 237.11214 g / mol and 237.11294 g / mol, respectively, with an error of -0.80 mDa for ([M+H+]+). The calculated and the found values were 259.09408 g / mol and 259.09532 g / mol, respectively, with an error of -1.24 mDa for ([M+Na+]+, as illustrated in FIG.3. Characterization of the compound of the formula (I)

[0049] Analysis of both1H- and13C-NMR spectra confirmed the formation of the compound of the formula (I). All NMR spectra were performed in d6-DMSO (99.0% atom, Across Organics, U.S.A.) using Bruker Biospin AG Magnet system 500MHz / 54mm instrument (Bruker BioSpin GmbH, Switzerland). Chemical shift in ppm were referenced to TMS as an internal reference. Data processing and analysis of the data were performed with topspin framework (Bruker Biospin GmbH, Version 3).

[0050] The1H-NMR data for thymoquinone (structure I) and doxorubicin (structure II), which are the main precursors of the compound of the formula (I), are summarized in Tables (1) and (2) to ease comparison with the spectrum of the compound of the formula (I).

[0004]

[0051] The1H-NMR spectrum of the compound is shown in FIG.4. As depicted from FIG. 4, all protons form the parent compounds appear in the spectrum, as in Table (3). The appearance of the proton of the thymoquinone ring is clear at ~6.5 ppm, and several additional signals in the aliphatic region. In addition, upon the formation of the compound, several CH2 can be observed using13C-dept 135-NMR at 19.1, 22.6, 29.4, 30.6, and 64.5 ppm corresponding to 1, 4, 6, b, and the succinic acid protons. However, some of the protons were hard to find due to the solvent signals, so the spectrum was also obtained in CD3OD. The CO-CH2-OH protons of CH2 appeared at 3.5 ppm and were directly connected to the CH2carbon at ~70 ppm. The assignments were made based on1H-1H and 1H-13C correlation NMR experiments. In addition, the amine group (NH2) in doxorubicin (appears at ~ 7.8 ppm and ~ 42.6 ppm for 1H and 15N atoms respectively) was converted to the amide group (-CO-NH-) which appeared at 7.5 ppm and 150.2 ppm for 1H and 15N atoms respectively, as shown in FIG.5.

[0052] The formation of thymoquinone-succinic acid-doxorubicin conjugate was confirmed using high resolution mass spectrometry. HRMS were acquired using electrospray ion trap ESI technique by collision-induced dissociation on a Bruker APEX- 4 (7 Tesla) instrument. The samples were infused using a syringe pump at a flow rate of 2 μL min−1.

[0053] HRMS (ESI) on the negative mode ([M–H+]−) confirmed thymoquinone-succinic acid-doxorubicin conjugate (C40H43NO14) synthesis, where the calculated and the found values were 761.26108 g / mol and 760.26023 g / mol, respectively, with an error of 0.85 mDa, as illustrated in FIG.6. thymoquinone-succinic acid-doxorubicin conjugate synthesis was also confirmed by the positive mode ([M+Na+]+) with calculated and found values were 784.25758 g / mol and 784.25892 g / mol, respectively, with an error of -1.43 mDa.

[0054] For purity detection of the compound of the formula (I), the compounds were analyzed using HPLC (Shimadzu, Japan). For HPLC analysis, C18 column has been used, the mobile phase composed of methanol: acetonitrile (60:40 volume ratio), the flow rate 1 ml / min and the detection wavelength at 254 nm. The HPLC spectrum of the compound of the formula (I) is illustrated in FIG.7. Example 3 Cytotoxicity Testing of the Compound of the Formula (I) on the Growth of Cancer and Normal Cells)

[0055] The compound of the present disclosure was then tested against human breast cancer cell lines (MCF-7) and compared to free thymoquinone and doxorubicin. Interestingly, the compound of the present disclosure showed high antitumor activity against tested cell lines (IC50 ~2 μM), as illustrated in FIG.8. Notably, the cytotoxic activity results of free doxorubicin and thymoquinone against MCF-7 cancer cells were (IC50 ~ 0.3 μM and 23 μM respectively), as illustrated in FIG.8 as well.

[0056] On the other hand, the compound of the present disclosure showed relative minimal cytotoxicity on normal cells including human-derived bone marrow cells and dermal fibroblasts (IC50 ~16 μM and ~10 μM respectively), compared to free doxorubicin which showed very high toxicity on these cells (IC50~0.2 μM and ~0.1 μM respectively), as illustrated in FIG. 9. These results clearly demonstrate that the compound of the formula (I) is capable to kill tumor cells, particularly invasive (ER)-positive and (PR)-positive breast adenocarcinoma, in high efficiency while exerted low toxicity on normal cells. Example 3 Subcellular Localization of the Compound of the Formula (I) in Cancer Cells )

[0057] Reference in this Example is being made to FIG. 10. To determine the internalization and subcellular localization of the compound of the present disclosure in comparison to free doxorubicin, an internalization assay was employed followed by Confocal Laser Scanning Microscopy (CLSM). Breast cancer cell line (MCF-7) was treated with either with the compound of the formula (I) or free doxorubicin and incubated at about 37oC for more than a 4-hour period as continuous uptake. Strikingly, the majority of molecules of the compound of the formula (I) at 4-hour uptake experiments were found to be located to perinuclear-like regions in the cytoplasm of cells, as illustrated in FIG.10. To further define such regions, they were co-stained with an ER marker. The results clearly show at both 1-hour and 4-hour uptake experiments, there is a high co-localization between the compound of the formula (I) and ER organelles (90%), indicating that the compound of the present disclosure is mainly localized to the ER region.

[0058] While embodiments of the present disclosure have been described in detail and with reference to specific embodiments thereof, it will be apparent to one skilled in the art that various additions, omissions, and modifications can be made without departing from the spirit and scope thereof.

[0059] In describing and claiming the present invention, the following terminology will be used.

[0060] The singular forms “a,” “an,” and “the” include plural referents unless the context clearly dictates otherwise.

[0061] As used herein, a plurality of items, structural elements, compositional elements, and / or materials may be presented in a common list for convenience. However, these lists should be construed as though each member of the list is individually identified as a separate and unique member. Thus, no individual member of such list should be construed as a defacto equivalent of any other member of the same list solely based on their presentation in a common group without indications to the contrary.

[0062] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood to one of ordinary skill in the art to which the presently disclosed subject matter belongs. Although any methods, devices, and materials similar or equivalent to those described herein can be used in the practice or testing of the presently disclosed subject matter, representative methods, devices, and materials are now described.

[0063] Unless otherwise indicated, all numbers expressing quantities of ingredients, reaction conditions, and so forth used in the specification and claims are to be understood as being modified in all instances by the term “about”. Accordingly, unless indicated to the contrary, the numerical parameters set forth in this specification and attached claims are approximations that can vary depending upon the desired properties sought to be obtained by the presently disclosed subject matter.

[0064] As used herein, the term “about”, when referring to a value or to an amount of mass, weight, time, volume, concentration or percentage is meant to encompass variations of in some embodiments ±20%, in some embodiments ±10%, in some embodiments ±5%, in some embodiments ±1%, in some embodiments ±0.5%, and in some embodiments ±0.1% from the specified amount, as such variations are appropriate to perform the disclosed method.

Claims

CLAIMS What is claimed is:

1. A compound of the formula (I), or a salt thereof:

2. A method for preparing the compound of claim 1, wherein the method comprises the steps of: Dissolving about thymoquinone and succinic acid in a mixture of acetonitrile and water under an inert atmosphere to provide a first mixture; Adding silver nitrate to the first mixture under the inert atmosphere at room temperature to provide a second mixture; Heating the second mixture under reflux, followed by a gradual addition of ammonium persulfate dissolved in distilled water to provide a third mixture; Diluting the third mixture with distilled water, followed by extracting the diluted third mixture using diethylether as an extraction solvent;Taking an organic layer resulting from the extraction step, drying it over anhydrous sodium sulfate powder, and evaporating such layer to produce thymoquinone-succinic acid conjugate; Dissolving thymoquinone-succinic acid conjugate in dimethyl sulfoxide (“DMSO”) while stirring under nitrogen gas to provide a first solution; Adding 1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide (“EDC”) to the first solution, followed by an addition of sulfo-NHS while stirring to provide a second solution; and Adding doxorubicin-HCl dissolved in DMSO with a trace amount of trimethylamine to the second solution with magnetic stirring in a dark place under nitrogen to produce the compound of the formula (I).

3. A pharmaceutical composition including a compound of formula (I) and / or pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier / excipient.

4. The pharmaceutical composition including a compound according to claim 3, wherein the composition is formulated as a solid, liquid, or semi-solid dosage form.

5. The pharmaceutical composition of claim 3, wherein the composition is administered via different routes such as oral, parenteral, intramuscular, intranasal, sublingual, intratracheal, ocular, vaginal, rectal, or intraventricular.

6. Use of the pharmaceutical composition of claims 3 to 5, for treating cancer.

7. The use according to claim 6, for treating breast cancer.

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