Pharmaceutical composition and use thereof for treating cancer

Abstract

There is provided a pharmaceutical composition for the treatment of cancer comprising a compound of general formula (I) or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable carrier. The prepared pharmaceutical composition is used as an HER2 selective inhibitor for treating many different types of HER2 amplification cancers, especially breast cancer. The pharmaceutical composition shows excellent activity to breast cancer cells, and is a safer and more effective alternative drug than a clinically used drug Herceptin. # imgabs0 #

Description

Technical Field

[0001] The present disclosure relates to pharmaceutical compositions for treating cancer, particularly for treating cancer that overexpresses HER2. Background of the Invention

[0003] Despite decades of basic and clinical research and trials of promising new therapies, cancer remains the second leading cause of death worldwide, with high morbidity and mortality, accounting for nearly 10 million deaths in 2020. The term "cancer" is primarily defined as a group of related diseases characterized by the development and growth of abnormal cells that can divide uncontrollably, thereby invading and destroying other parts of the body.

[0004] In cancer research, the key role of human epidermal growth factor receptor 2 (HER2) in tumorigenesis has been widely recognized. HER2 is a transmembrane tyrosine kinase receptor protein that acts as a mediator of cell growth, differentiation and survival. However, HER2 gene amplification and receptor overexpression can lead to the development of many types of human cancers, including breast cancer, colon cancer, endometrial cancer, cervical cancer, urothelial cancer, lung cancer, ovarian cancer, gastric cancer, gastroesophageal junction cancer, head and neck cancer, biliary tract cancer, prostate cancer and pancreatic cancer. It is worth noting that HER2 is highly expressed in about 5-20% of breast cancer patients and serves as an important marker for poor prognosis.

[0005] The relationship between abnormal HER2 expression and cancer is of great significance in cancer treatment. Therefore, HER2 has been identified as an ideal biological target for the treatment of cancer, especially breast cancer. This is because the formation and spread of cancer cells can be limited by inhibiting HER2. Therefore, many therapeutic strategies are aimed at the complex mechanism of regulating HER2 function to selectively inhibit the growth of HER2-overexpressing cancer cells. In addition, many studies have focused on ligand-based and structure-based studies to evaluate the efficacy of existing therapeutic agents or compounds as potential HER2 inhibitors. For example, a study entitled "Design, synthesis, and antitumor activity of new bis-aminomethylnapthalenes" published by Mariela Bollini et al. in 2008 disclosed a method for preparing a series of bis-aminomethylnaphthalenes by a simple synthetic strategy using reductive amination. The anticancer activity of the synthetic compounds and their ability to act as DNA binding ligands were evaluated. None of the prior art schemes discloses these synthetic derivatives, especially the general formula C 30 H 37Use of (6-methoxy-naphthalen-2-ylmethyl)-[2-(2-{2-[(6-methoxy-naphthalen-2-ylmethyl)-amino]-ethoxy}-ethoxy)-ethyl]-amine of ClN2O4 as a potential HER2 inhibitor for the treatment of cancer. SUMMARY OF THE INVENTION

[0007] Therefore, the object of the present disclosure is to provide a pharmaceutical composition comprising a compound of general formula (I) or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable carrier for use in treating cancer:

[0008]

[0009] In aspects of the present disclosure, the pharmaceutical composition may be formulated as a solid, liquid, or semisolid dosage form.

[0010] In some aspects, the pharmaceutical composition can be administered by various routes, such as oral, parenteral, intramuscular, intranasal, sublingual, intratracheal, ocular, vaginal, rectal, or intraventricular.

[0011] In some aspects, the pharmaceutical composition may further include one or more therapeutic agents as a combination therapy.

[0012] In aspects of the present disclosure, the pharmaceutical composition can be used to treat different types of HER2 amplified cancers.

[0013] In some aspects, the different types of HER2 amplified cancers can be breast cancer, colon cancer, endometrial cancer, cervical cancer, urothelial cancer, lung cancer, ovarian cancer, gastric cancer, gastroesophageal junction cancer, head and neck cancer, biliary tract cancer, prostate cancer, and pancreatic cancer.

[0014] BRIEF DESCRIPTION OF THE DRAWINGS

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

[0016] Figure 1 Shown is a line graph of tumor growth versus time in mice treated with different doses of a pharmaceutical composition prepared according to embodiments of the present disclosure.

[0017] Figure 2 A bar graph showing tumor weights obtained from mice after treating the mice for 4 weeks with different doses of pharmaceutical compositions prepared according to embodiments of the present disclosure.

[0018] Figure 3 A bar graph showing the percent inhibition of tumor growth in mice after treatment of the mice for 4 weeks with different doses of a pharmaceutical composition according to an embodiment of the present disclosure.

[0019] Details

[0020] Embodiments of the present disclosure provide a pharmaceutical composition comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable carrier. In some embodiments, the pharmaceutical composition may further comprise one or more therapeutic agents as a combination therapy.

[0021]

[0022] In embodiments of the present disclosure, the composition may be formulated as a solid, liquid, or semisolid dosage form.

[0023] In some embodiments of the present disclosure, the composition can be administered by various routes, such as oral, parenteral, intramuscular, intranasal, sublingual, intratracheal, ocular, vaginal, rectal, or intraventricular routes.

[0024] Embodiments of the present disclosure provide uses of the pharmaceutical composition for treating different types of HER2 amplified cancers. In some embodiments of the present disclosure, the pharmaceutical composition may act as a selective HER2 inhibitor.

[0025] In some embodiments of the present disclosure, the different types of HER2 amplified cancers may be breast cancer, colon cancer, endometrial cancer, cervical cancer, urothelial cancer, lung cancer, ovarian cancer, gastric cancer, gastroesophageal junction cancer, head and neck cancer, biliary tract cancer, prostate cancer, and pancreatic cancer.

[0026] The present disclosure is now further described according to the examples and detailed description, from which additional features and advantages can be obtained. It should be noted that the following explanations are provided for the purpose of illustration and description only; they are not intended to be exhaustive or limit the present disclosure to the precise form disclosed. Example

[0027] Example 1

[0028] Preparation of pharmaceutical compositions

[0029] The pharmaceutical compositions of the present disclosure comprising the compounds of general formula (I) were synthesized with reference to previously published methods with minor modifications.

[0030] The synthesis of the compounds started with the formation of bis-imines followed by reduction of these imines with NaBH4 in methanol to give the desired compounds of general formula C 30 H 37The compound (6-methoxy-naphthalen-2-ylmethyl)-[2-(2-{2-[(6-methoxy-naphthalen-2-ylmethyl)-amino]-ethoxy}-ethoxy)-ethyl]-amine of ClN2O4 was obtained in good yield. The compound was then precipitated by HCl. All required characterizations (NMR and 2D NMR and mass spectrometry) were also performed to accurately determine and ensure the structure of the desired compound and its purity.

[0031] Then, through molecular modeling and the National Cancer Institute,

[0032] Computer screening of the NCI (National Cancer Institute) database identified the compound as a potential HER2 inhibitor.

[0033] Example 2

[0034] In vitro testing of pharmaceutical compositions

[0035] Antiproliferative experiments were performed to evaluate the effects of the compound on different normal and breast cancer cell lines with different HER2 expression levels. The compound was tested in vitro on three different breast cancer cell lines, namely SKBR3 cells (overexpressing HER2 receptors), MDA 231 cells (cells that do not express HER2) and MCF7 cells (cells that do not express HER2). Normal non-cancerous cells were normal skin fibroblasts, normal breast cells MCF10 and cardiomyocytes. The compound showed the best activity and selectivity against HER2 overexpressing cells (SKBR3) compared to normal non-cancerous cells.

[0036] Example 3

[0037] Mechanism of action of the drug combination

[0038] The use of the prepared compound for treating cancer, especially cancer cells overexpressing HER2, lies in its mechanism of action. The mechanism of action of the compound was studied using immunoblotting technology. The results of this technology clearly demonstrated that the compound can target the HER2 receptor by binding to the ATP binding pocket of the intracellular kinase domain of the HER2 receptor, thereby inhibiting the phosphorylation of HER2. When the phosphorylation of HER2 is blocked, its activation and downstream signaling pathways that promote cancer cell proliferation are also inhibited.

[0039] The half maximal inhibitory concentration (IC) of the compounds on SKBR3, MCf10 and fibroblasts 50 ) were 450, 2500 and 2000 nM respectively.

[0040] Example 4

[0041] The median lethal dose (LD) of the drug composition 50 )

[0042] The acute toxicity of a drug can be determined by calculating its median lethal dose (LD 50 ) was determined. Repeated dose toxicity studies were conducted by intraperitoneal administration of compounds of the disclosure in 36 male albino mice, followed by a recovery period. The intraperitoneal route was chosen because it is the most likely human exposure route in the environment. This study was based on internationally recognized guidelines, and the dose levels selected for the toxicity studies were based on the initial in vitro cytotoxicity studies that were set.

[0043] The approximate LD was initially determined by the so-called “step method” using a small number of mice (2 per dose) and increasing drug doses. 50 As a pilot study, five doses (250, 200, 150, 100, and 50 mg / kg body weight) were selected and administered intraperitoneally to six groups of mice (two mice in each group) to determine the LD 50 The mice were observed for any symptoms of poisoning at the first 2 hours and then at the 6th and 24th hours. After 24 hours, the number of dead mice in each group was counted. Table 1 below shows the mortality rate at different doses of the tested compounds:

[0044] Table (1)

[0045]

[0046] The percentage mortality rate at each dose level is provided by the following calculation and converted to probability units:

[0047] For 0% mortality:

[0048] For 100% mortality:

[0049] Where n is the number of mice in each group. Then plot the probability unit value against the logarithm of the dose. Determine the dose corresponding to probability unit 5, that is, 50% mortality. The result determines the LD 50 It is 125mg / Kg body weight.

[0050] Example 5

[0051] In vivo testing of pharmaceutical compositions

[0052] Adenocarcinoma tumors (SKBR3) were inoculated orthotopically into the mammary pads of approximately 40 4-6 week-old athymic NCRnu / nu nude mice weighing approximately 23±2 g to study the antitumor activity of the compounds. After two weeks of tumor growth, the mice were randomly divided into 5 groups, each containing 6 mice. The pharmaceutical composition was administered orally by gavage (wt / wt) at a dose of 60 mg / kg body weight (Group III), 40 mg / kg body weight (Group IV), and 20 mg / kg body weight (Group V), once a day for 4 weeks. Mice in the negative control group (Group I) received only 0.2 ml of distilled water, while mice in the positive control group (Group II) were injected intraperitoneally with 15 mg / kg Herceptin twice a week. Tumor size and body weight were recorded before the start of treatment and every week. Tumor size was calculated by applying the following formula:

[0053]

[0054] Where L is the length of the tumor measured with a caliper, W is its width, and D is its depth.

[0055] At the end of the experiment, mice were euthanized with CO2 and cervical dislocation. Their xenograft tumors were collected and cut into two parts. The first half of the tumor was preserved in 4% paraformaldehyde for histopathological studies, vascularization, histomorphology, and apoptosis. The other half of the tumor was stored at -80°C until further evaluation of cell cycle parameters using flow cytometry.

[0056] In this embodiment, reference Figure 1-3 . Figure 1 A line graph showing the tumor growth curves in all test mouse groups after treatment with different doses of the compound for a period of 6 weeks. Compared with the negative control group (Group I), tumor growth was inhibited, and there was almost no change in tumor growth over time as the dose of the compound increased. Figure 2 A bar graph is shown showing the weight of the tumors obtained from each test mouse group after 4 weeks of treatment with different doses of the compound. The mice in group I that did not receive treatment showed the largest tumor size, followed by mice treated with 20 mg / kg body weight (group III), 40 mg / kg body weight (group IV), 15 mg / kg Herceptin (group II) and 60 mg / kg body weight (group V), respectively. Thus, it can be noted that administration of the compound at a dose of 60 mg / kg body weight showed a more pronounced tumor inhibitory effect than the other doses tested. Figure 3A bar graph showing the percentage inhibition of breast tumors after 4 weeks of treatment of all test mouse groups with different doses of compound. Compounds at 20, 40 and 60 mg / kg concentrations showed dose-dependent inhibition of SKBR3 tumor growth with inhibition rates of 46.3, 62.9 and 75.9%, respectively. Positive control group II treated with 15 mg / kg Herceptin showed a 63.8% reduction in breast tumors compared to the negative control group (group I).

[0057] The results of this in vivo analysis showed clear evidence that the compound has potent antitumor activity against breast adenocarcinoma (SKBR3), thus selecting the drug as an effective and potential anti-HER2 agent. In addition, the activity and selectivity of the compound against breast cancer models were superior to the clinically used drug Herceptin, which is expensive and has many side effects.

[0058] While the embodiments of the present disclosure have been described in detail and with reference to specific embodiments thereof, it will be apparent to those 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 is used.

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

[0061] As used herein, for convenience, multiple items, structural elements, compositional elements and / or materials may be presented in a common list. However, these lists should be interpreted as each member of the list being individually identified as a separate and unique member. Therefore, in the absence of a contrary indication, any single member of such a list should not be interpreted as a de facto equivalent of any other member of the same list.

[0062] As used herein, the term "about," when referring to a value or amount of mass, weight, time, volume, concentration, or percentage, is meant to include variations from the specified amount of ±20%, in certain embodiments ±10%, in certain embodiments ±5%, in certain embodiments ±1%, in certain embodiments ±0.5%, and in certain embodiments ±0.1%, as such variations are suitable for performing the disclosed methods.

[0063] As used in the present invention, the term "treatment" or "treatment" refers to reducing the severity of HER2-amplified cancer, delaying the occurrence of HER2-amplified cancer, slowing the growth of HER2-amplified cancer, alleviating the cell metastasis of HER2-amplified cancer, shortening the duration of HER2-amplified cancer, and hindering the development of HER2-amplified cancer. It includes causing cancer regression, alleviating the symptoms caused by HER2-amplified cancer, or stopping the symptoms caused by HER2-amplified cancer. The term "treatment" or "treatment" includes but is not limited to preventive and / or therapeutic treatment or treatment.

Claims

1. A pharmaceutical composition comprising a compound of general formula (I) or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable carrier.

2. The pharmaceutical composition of claim 1, wherein the pharmaceutical composition is formulated into a solid, liquid or semisolid dosage form.

3. The pharmaceutical composition of claim 1, wherein the pharmaceutical composition is administered by different routes, including oral, parenteral, intramuscular, intranasal, sublingual, intratracheal, ocular, vaginal, rectal or intraventricular routes.

4. The pharmaceutical composition of claim 1, wherein the pharmaceutical composition further comprises one or more therapeutic agents as a combination therapy.

5. Use of the pharmaceutical composition of claim 1 for treating different types of HER2 amplified cancers.

6. Use of the pharmaceutical composition of claim 5, wherein the different types of HER2-amplified cancers are breast cancer, colon cancer, endometrial cancer, cervical cancer, urothelial cancer, lung cancer, ovarian cancer, gastric cancer, gastroesophageal junction cancer, head and neck cancer, biliary tract cancer, prostate cancer and pancreatic cancer.

Images