Composition, method and use thereof for delaying primary tumor metastasis using lomitapide

Abstract

The present disclosure provides a method for delaying preliminary tumor metastasis. The method involves: (a) preparing a composition comprising Lomitapide and or a pharmaceutically acceptable salt, ester, solvate, analogue, thereof or in combination with one or more therapeutic compound; and (b) administering a therapeutically effective amount of the composition, typically in the range of 50-70 mg, to a subject in need. The disclosure particularly pertains to the use of such compositions for delaying tumor metastasis, with Lomitapide.

Description

[0001] COMPOSITION, METHOD AND USE THEREOF FOR DELAYING PRIMARY TUMOR METASTASIS USING LOMITAPIDE

[0002] TECHNICAL FIELD

[0003] The present disclosure relates to a method of delaying primary tumor metastasis. The present disclosure also relates to the composition and more particularly to for the treatment of for delaying primary tumor metastasis.

[0004] BACKGROUND

[0005] Metastasis represents a critical and complex aspect of cancer progression, involving the dissemination of cancer cells from the original tumor site to distant organs through the bloodstream or lymphatic system. This multi-step process includes detachment of cells from the primary tumor, invasion of nearby tissues, entry into the circulatory or lymphatic system, survival during transit, and establishment of new tumors in other tissues. Cellular mechanisms, such as epithelial-to- mesenchymal transition (EMT) to boost mobility and mesenchymal-to-epithelial transition (MET) to support colonization, play pivotal roles.

[0006] As a leading cause of cancer-related deaths, metastasis poses significant challenges for treatment, particularly in colorectal cancer where the liver and lungs are frequent secondary targets. Despite notable advancements in cancer therapies, effective strategies specifically targeting metastasis remain scarce, hindered by the complexity of the process and obstacles in clinical trials.

[0007] The onset of metastasis, referred to as "primary metastasis delay," varies based on tumor type, biological characteristics, and host environment. Prolonged treatment delays for metastatic cancer have been linked to poorer survival outcomes, with a six-month delay associated with a higher risk of localized tumor progression and distant metastasis.

[0008] Research emphasizes the critical need to prevent or slow metastasis to improve patient survival and prognosis. Steps like intravasation into blood or lymphatic vessels, survival in circulation, and extravasation into distant tissues highlight the intricate journey of metastatic cells. EMT enhances cellular mobility and invasiveness, while MET supports secondary tumor establishment. Metastasis remains the primary driver of cancer mortality and presents limited treatment options. The clinical implications of metastasis are profound, as it is the primary cause of cancer-related mortality. Metastatic disease is often associated with poorer outcomes and limited treatment options, underscoring the urgent need for more effective therapeutic strategies targeting metastatic processes. The US Food and Drug Administration (FDA) approved 124 anticancer drugs for 255 solid tumor indications between 2003 and 2021. While these drugs significantly reduced the risk of death by 27% and tumor progression by 43%, the median extension of both overall survival (OS) and progression-free survival (PFS) was limited to 2.80 and 3.30 months, respectively. The primary reason behind this limitation is the prevailing absence of treatments capable of impeding the metastatic process, which accounts for the demise of 90% of cancer patients.

[0009] Additionally, the failure of several clinical candidates aimed at targeting metastasis has raised questions about the translational relevance of the target-based approach. Ironically, many research initiatives focusing on proliferation targets neglect to assess the target's potential impact on impeding metastasis. Consequently, they miss out on candidates that could potentially slow down metastasis and serve as primary treatment options.

[0010] Current cancer treatments address cell proliferation, immune responses, and targeted inhibition but lack focus on delaying or halting metastasis. Failures in targeting metastasis have raised questions about the translational relevance of current approaches, often overlooking candidates that could impede metastatic spread.

[0011] Addressing these gaps is crucial, particularly in introducing metastasis- targeted therapies as primary treatments in adjuvant or neo-adjuvant settings. Such therapies must ensure safety for chronic use by patients with primary tumors.

[0012] The complexities surrounding metastasis necessitate urgent exploration of novel therapeutic strategies that not only impede cancer spread but also improve long- term outcomes for patients. Efforts to identify safe and effective molecules that specifically address primary metastasis are essential for advancing cancer care.

[0013] Hence, there is a definite need for identifying clinical candidates or molecules that can specifically impede primary metastasis as well as are safe to administer to cancer patients.

[0014] SUMMARY

[0015] In an aspect of the invention, a method for delaying tumor metastasis, comprising: a. preparing a composition comprising Lomitapide and or a pharmaceutically acceptable salt, ester, solvate, analogue, thereof or in combination with one or more therapeutic compound; and b. administering the therapeutic effective amount of the obtained composition in the range of 50-70 mg, to a subject in need thereof.

[0016] In yet another aspect of the present disclosure, in the method using the composition further comprises a pharmaceutically acceptable carrier, excipient, diluent and / or adjuvant.

[0017] In further aspect of the present disclosure, in the method using the composition, the pharmaceutical salts are selected from the group comprising mesylate, maleate, and camsylate.

[0018] In another aspect of the present disclosure, in the method using the composition, pharmaceutically acceptable carrier selected from the group comprising solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like, compatible with pharmaceutical administration.

[0019] In another aspect of the present disclosure, in the method using the composition, the therapeutic compound is selected from the group comprising losartan, propranolol, metformin, aspirin, atenolol, niclosomide and combinations thereof.

[0020] In another aspect of the present disclosure, in the method, the composition, comprises Lomitapide and therapeutic compound in a ratio of 1:99. In another aspect of the present disclosure, in the method, the composition comprises Lomitapide and therapeutic compound in a ratio of 99: 1.

[0021] In another aspect of the present disclosure, in the method using the composition, the composition is administered in the form of tablets, troches, or capsules.

[0022] In an aspect of the present disclosure, the method is used when the subject has been diagnosed from the group comprising colorectal cancer, head and neck cancers, triple-negative breast cancer, pancreatic cancer, liver cancer, ovarian cancer, esophageal cancer, cervical cancer, lung cancer, bladder cancer, and kidney cancer. In another aspect of the present disclosure, in the method using the composition, the composition is administered as two, three, four, five, six or more sub-doses can be administered separately at appropriate intervals throughout the day.

[0023] In another aspect of the present disclosure, in the method using the composition, concentration of Lomitapide in the composition may range from as low as 0.1% of the total amount of the composition up to as high as 100%.

[0024] In another aspect of the present disclosure, in the method using the composition, the dosage of the composition is in the range of 50-70 mg per person per day.

[0025] In another aspect of the present disclosure, the method uses composition that comprises 50-65 mg of Lomitapide, pharmaceutically acceptable salts and therapeutic compounds.

[0026] In another aspect of the present disclosure, in the method the composition administered preferably comprises 60 mg of Lomitapide.

[0027] In another aspect of the present disclosure, in the method, the composition of delaying tumor metastasis comprises Lomitapide and or a pharmaceutically acceptable salt, ester, solvate, analogue, thereof or in combination with one or more therapeutic compound.

[0028] In another aspect of the present disclosure, the composition further comprises a pharmaceutically acceptable carrier, excipient, diluent and / or adjuvant. In another aspect of the present disclosure, the therapeutic compound is selected from the group comprising losartan, propranolol, metformin, aspirin, atenolol, niclosomide and combinations thereof.

[0029] In another aspect of the present disclosure, the pharmaceutical salts are selected from the group comprising mesylate, maleate, and camsylate.

[0030] In another aspect of the invention, the composition comprises Lomitapide and therapeutic compound in a ratio of 1:99.

[0031] In another aspect of the present disclosure, pharmaceutically acceptable carrier selected from the group comprising solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like, compatible with pharmaceutical administration.

[0032] In another aspect of the present disclosure, the composition comprises 50-65 mg of Lomitapide, pharmaceutically acceptable salts and therapeutic compounds.

[0033] In another aspect of the present disclosure, in the method the composition preferably comprises 60 mg of Lomitapide.

[0034] In an aspect of the present disclosure, a composition for the treatment of delaying tumor metastasis comprising: Lomitapide and or a pharmaceutically acceptable salt, ester, solvate, analogue, thereof or in combination with one or more therapeutic compound.

[0035] In yet another aspect of the present disclosure, the composition for treating delaying tumor metastasis comprises 50-65 mg of Lomitapide, pharmaceutically acceptable salts and therapeutic compounds.

[0036] In further aspect of the present disclosure, the composition for treating delaying tumor metastasis preferably comprises 60 mg of Lomitapide.

[0037] In some other aspect of the present disclosure, the composition is a cocktail with Lomitapide and another therapeutic compound.

[0038] In some other aspect of the present disclosure, in the composition the lomitapide may inhibit tumor cell extravasation by at least about 50% compared to a control. In some other aspect of the present disclosure, a method for inhibiting tumor cell transendothelial migration (TEM) in a subject with cancer is provided.

[0039] In some other aspect of the present disclosure, the method includes administering to the subject an effective amount of lomitapide.

[0040] In some other aspect of the present disclosure, the lomitapide may inhibit tumor cell transendothelial migration by at least about 90% compared to a control.

[0041] In some other aspect of the present disclosure, a method for increasing hypoxia response with colorectal cancer in a subject is provided. The method includes administering to the subject an effective amount of therapeutic compound.

[0042] In another aspect of the invention, the lomitapide may increase vimentin expression in the colorectal cancer cells in present method. The lomitapide may decrease e- cadherin expression in the colorectal cancer cells in the present method.

[0043] In some other aspect of the present disclosure, use of lomitapide for the manufacture of a medicament for treating metastasis in a subject with colorectal cancer is provided.

[0044] In some other aspect of the present disclosure, the medicament may be for oral administration. The medicament may be formulated for administration at a dose of about 50 mg to about 70 mg per day.

[0045] In some other aspect of the present disclosure, use of lomitapide for the manufacture of a medicament for inhibiting tumor cell extravasation in a subject with colorectal cancer is provided.

[0046] In another aspect of the present disclosure, clinical candidate that is safe to be administered to cancer patients.

[0047] The foregoing general description of the illustrative embodiments and the following detailed description thereof are merely exemplary aspects of the teachings of this disclosure and are not restrictive.

[0048] BRIEF DESCRIPTION OF DRAWINGS

[0049] The above and still further features and advantages of aspects of the present disclosure becomes apparent upon consideration of the following detailed description of aspects thereof, especially when taken in conjunction with the accompanying drawings, and wherein:

[0050] FIG. 1 depicts a K-means clustering heat map of top 10 approved drugs identified using a proprietary algorithm, tested across the weighted assays, in accordance with an aspect of the present disclosure;

[0051] FIG. 2A depicts a dot plot analysis of EMT assay depicting effect of ImM Lomitapide treatment on HT29 cells, in accordance with an aspect of the present disclosure;

[0052] FIG. 2B depicts the effect of Lomitapide treatment on HT29 cells by showing the change in Ecad and Vim populations and the change in plasticity ratio (PR), in accordance with an aspect of the present disclosure;

[0053] FIG. 3A depicts a dot plot analysis of MET assay depicting effect of ImM Lomitapide treatment on SW480 cells, in accordance with an aspect of the present disclosure;

[0054] FIG. 3B depicts the effect of Lomitapide treatment on SW480 cells by showing the change in Ecad and Vim populations and the change in plasticity ratio (PR), in accordance with an aspect of the present disclosure;

[0055] FIG. 4A depicts a graph depicting trans-endothelial cell migration assay of tumor cells (treated with 1 pM Lomitapide) across HUVEC cells, in accordance with an aspect of the present disclosure;

[0056] FIG. 4B depicts a graph depicting Extravasation assay of tumor cells (treated with 1 pM Lomitapide), HUVEC cells and Platelet-rich plasma (PRP), in accordance with an aspect of the present disclosure;

[0057] FIG. 5A depicts a graph depicting % positive population of SW480 cells treated with DMSO control, Lomitapide and HIF la inhibitor IDF-11774(IDF-11774- HIF1 a inhibitor), In accordance with an aspect of the present disclosure;

[0058] FIG. 5B depicts a graph depicting total colony count of SW480 cells after treating with DMSO, Retinoic acid, Retinoic acid and AGN (AGN193109 is a MET inhibitor), AGN only, and Lomitapide, respectively, in accordance with an aspect of the present disclosure;

[0059] FIG. 6 depicts a graph depicting effect of Lomitapide on senescence using primary patient cells, in accordance with an aspect of the present disclosure;

[0060] FIG. 7 depicts a graph depicting effect of Lomitapide on metastasis in the larval stage of Drosophila, moving from day 5 to day 8, in accordance with an aspect of the present disclosure;

[0061] FIG. 8A depicts a flow chart of animal model experimentation; in accordance with an aspect of the present disclosure;

[0062] FIG. 8B depicts a graph depicting effect of 5 and 10 mg / kg Lomitapide on primary tumor size, in accordance with an aspect of the present disclosure;

[0063] FIG. 8C depicts a graph showing the effect of 5 and 10 mg / kg Lomitapide, wherein the graph depicts effect on lung metastasis; and

[0064] FIG. 8D depicts a graph showing the effect of 5 and 10 mg / kg Lomitapide, wherein, graph depicts effect on liver metastasis, in accordance with an aspect of the present disclosure.

[0065] Fig 9A depicts a graph depicting effect of 1, 3.5 and 12.3 mg / kg on primary tumor size, in accordance with an aspect of the present disclosure;

[0066] Fig. 9B depicts a graph depicting the tumour tissue levels of Lomitapide in ng / ml, in accordance with an aspect of the present disclosure;

[0067] Fig. 9C depicts a graph depicting effect of 1, 3.5 and 12.3 mg / kg on the body weight of mice, in accordance with an aspect of the present disclosure;

[0068] Fig. 10A depicts a graph showing the effect of 1, 3.5 and 12.3 mg / kg Lomitapide on lung metastasis,

[0069] Fig. 10B depicts a graph depicting the plasma levels of Lomitapide in ng / ml, in accordance with an aspect of the present disclosure; and

[0070] Fig. 10 C depicts a graph showing the effect of 1, 3.5 and 12.3 mg / kg Lomitapide on liver metastasis. To facilitate understanding, reference numerals have been used, where possible, to designate elements common to the figures.

[0071] DETAILED DESCRIPTION

[0072] Various aspects of the present disclosure provide an information processing apparatus for analysis of a biological sample, system and a method thereof. The following description provides specific details of certain aspects of the disclosure illustrated in the drawings to provide a thorough understanding of those aspects. It should be recognized, however, that the present disclosure can be reflected in additional aspects and the disclosure may be practiced without some of the details in the following description.

[0073] The words / phrases "exemplary", “example”, “illustration”, “in an instance”, “and the like”, “and so on”, “etc.”, “etcetera” are merely used herein to mean "serving as an example, instance, or illustration." Any embodiment or implementation of the present subject matter described herein using the words / phrases "exemplary", “example”, “illustration”, “in an instance”, “and the like”, “and so on”, “etc ”, “etcetera” is not necessarily to be construed as preferred or advantageous over other embodiments.

[0074] The term “analogue” is typically used to denote a compound that has a chemical structure that is substantially similar to the structure of the parent compound, whilst retaining at least some of the biological function of the parent compound.

[0075] The term "pharmaceutically acceptable salts" typically refers to salts prepared from pharmaceutically acceptable substantially nontoxic bases or acids including inorganic or organic bases and inorganic or organic acids, as well as salts that can be converted into pharmaceutically acceptable salts.

[0076] The term “a therapeutically effective amount”, as used herein typically refers to an amount of a compound that may be used is sufficient to effect beneficial or desired results as described herein when administered to a subject such as a mammal, preferably a human, in need of such therapy; for example, a subject who is suffering from cancer. The term "pharmaceutically acceptable carrier" typically includes solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like, compatible with pharmaceutical administration.

[0077] Lomitapide (sold under the brand names Jaxtapid (US) and Lojuxta (EU)) is an orphan drug used as a lipid-lowering agent for the treatment of familial hypercholesterolemia, developed by Aegerion Pharmaceuticals. The US Food and Drug Administration (FDA) approved lomitapide in Dec. 21, 2012, as an orphan drug to reduce EDU cholesterol, total cholesterol, apolipoprotein B, and non-high- density lipoprotein (non-HDL) cholesterol in people with homozygous familial hypercholesterolemia. Homozygous familial hypercholesterolemia is a rare genetic disorder of low-density lipoprotein cholesterol (LDL-C) metabolism resulting in extremely elevated serum levels of LDL-C and premature atherosclerotic cardiovascular disease.

[0078] Lomitapide has already been established as a safe drug to treat familial hypercholesterolemia in humans, as long-term administration of Lomitapide resulted in no complications such as spontaneous cancer development. Further, the side effect profile of Lomitapide is generally more favorable than that of most drugs typically used to treat cancer.

[0079] Previous studies have primarily concentrated on inhibiting the migration and invasion stages of cancer progression. These studies typically involved screening for compounds capable of inhibiting either or both of these processes.

[0080] Using their proprietary platforms and algorithms, the inventors of the present invention identified possible non-oncology drugs that may facilitate in delaying primary tumor metastasis. Surprisingly, the inventors identified that Lomitapide hinders metastasis. Following extensive experimentation and analysis, the inventors of the present invention have identified that Lomitapide migrates cells more towards the mesenchymal axis, thereby increasing Plasticity Ratio (PR) substantially. A high PR theoretically leads to higher metastasis. Lomitapide was also observed to increase senescence in patient's primary tumor cells, thereby diminishing their re-epithelialization chances. Furthermore, Lomitapide was found to inhibit trans endothelial migration (TEM), extravasation, upregulate HIFla levels, prevent colony formation and also completely inhibit metastasis in larval stage of Drosophila.

[0081] From these observations the inventors have summarized that Lomitapide can extend survival by delaying metastasis, as it delays the proliferation of primary tumor cells by making them disseminate cells but does not allow those seeded cells to enter and regrow in the secondary tissue. In the secondary tissue, Lomitapide does not promote mesenchymal to epithelial transition. Instead, it shifts cells more to the mesenchymal axis, significantly increasing the PR. This increase in PR maintains cells in their mesenchymal axis and abrogates secondary tumor colonization and also increases senescence.

[0082] In a study using Drosophila animal model, the inventors of the present invention observed that Lomitapide significantly reduces metastasis, without significantly reducing primary tumor size. In another animal model study, the inventors of the present invention observed that Lomitapide significantly reduces metastasis in the liver and lung without having a similar statistically significant reduction in the affecting the primary tumor growth.

[0083] In an embodiment, a method for delaying tumor metastasis, comprising: c. preparing a composition comprising Lomitapide and or a pharmaceutically acceptable salt, ester, solvate, analogue, thereof or in combination with one or more therapeutic compound; and d. administering the therapeutic effective amount of the obtained composition in the range of 50-70 mg, to a subject in need thereof.

[0084] In yet another embodiment, in the method, the prepared composition further comprises a pharmaceutically acceptable carrier, excipient, diluent and / or adjuvant. In further embodiment, in the method, the prepared composition further comprises the pharmaceutical salts are selected from the group comprising mesylate, maleate, and camsylate. In another embodiment, in the method, the prepared composition further comprises the pharmaceutically acceptable carrier selected from the group comprising solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like, compatible with pharmaceutical administration.

[0085] In another embodiment, in the method, the prepared composition further comprises the therapeutic compound selected from the group comprising losartan, propranolol, metformin, aspirin, atenolol, niclosomide and combinations thereof.

[0086] In another embodiment, in the method, the composition comprises Lomitapide and therapeutic compound in a ratio of 1:99.

[0087] In another embodiment, in the method, the composition comprises Lomitapide and therapeutic compound in a ratio of 99: 1.

[0088] In another embodiment, in the method, administering the composition is in the form of tablets, troches, or capsules.

[0089] In another embodiment, the method is used when the subject has been diagnosed from the group comprising colorectal cancer, head and neck cancers, triple-negative breast cancer, pancreatic cancer, liver cancer, ovarian cancer, Esophageal cancer, cervical cancer, lung cancer, bladder cancer, and kidney cancer.

[0090] In another embodiment, in the method, the composition may be administered as two, three, four, five, six or more sub-doses, separately at appropriate intervals throughout the day.

[0091] In another embodiment, in the method, the concentration of Lomitapide in the composition may range from as low as 0.1% of the total amount of the composition up to as high as 100%.

[0092] In another embodiment, in the method, the concentration of Lomitapide in the composition is from 1% to 90% by weight.

[0093] In another embodiment, in the method, the concentration of Lomitapide in the composition preferably in the range from 5% to 80% by weight. In another embodiment, in the method, the concentration of Lomitapide in the composition more preferred from 10% to 70% by weight.

[0094] In another embodiment, in the method the dosage of the composition administered is in the range of 50-70 mg, to a subject in need thereof.

[0095] In another embodiment, in the method, the composition administered comprises 5- 60 mg of Lomitapide, pharmaceutically acceptable salts and therapeutic compounds.

[0096] In another embodiment, in the method, the composition is in the form of an oral formulation.

[0097] In an embodiment, the present disclosure provides a composition of delaying tumor metastasis comprising Lomitapide and or a pharmaceutically acceptable salt, ester, solvate, analogue, thereof or in combination with one or more therapeutic compound.

[0098] In yet another embodiment, the present disclosure provides the composition that further comprises a pharmaceutically acceptable carrier, excipient, diluent and / or adjuvant.

[0099] In further embodiment, the pharmaceutical salts in the composition are selected from the group comprising mesylate, maleate, and camsylate.

[0100] In another embodiment, the therapeutic compound in the composition is selected from the group comprising losartan, propranolol, metformin, aspirin, atenolol, niclosomide and combinations thereof.

[0101] In another embodiment, the composition comprises lomitapide and therapeutic compound in a ratio of 1:99.

[0102] In another embodiment, pharmaceutically acceptable carrier in the composition is selected from the group comprising solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like, compatible with pharmaceutical administration.

[0103] In an embodiment, a composition for the treatment of delaying tumor metastasis comprising: Lomitapide and or a pharmaceutically acceptable salt, ester, solvate, analogue, thereof or in combination with one or more therapeutic compound is provided.

[0104] In yet another embodiment, the composition for treating delaying tumor metastasis comprises 50-65 mg of Lomitapide, pharmaceutically acceptable salts and therapeutic compounds.

[0105] In further embodiment, the composition for treating delaying tumor metastasis preferably comprises 60 mg of Lomitapide.

[0106] In another embodiment, the composition for treating delaying tumor metastasis includes Lomitapide either alone, or in combination with one or more therapeutic compound.

[0107] In another embodiment, the dosage of composition that may be used in the present inventions will vary with the route of administration, the rate of excretion, the duration of the treatment, the identity of any other therapeutic compounds being administered, the age, size, and species of the subject, e.g., human patient, and like factors. In general, the dosage of a compound that may be used in the present invention will be an amount which is the lowest dose effective to produce the desired effect with no or minimal side effects.

[0108] In another embodiment, the composition may further include one or more therapeutic compound. Non-limiting examples of therapeutic compound that may be used for delaying primary tumor metastasis include losartan, propranolol, metformin, aspirin, atenolol, niclosomide and combinations thereof.

[0109] In yet another embodiment, the use of the composition for treating delaying tumor metastasis comprises 50-65 mg of Lomitapide, pharmaceutically acceptable salts and therapeutic compounds.

[0110] In further embodiment, the use of composition for treating delaying tumor metastasis preferably comprises 60 mg of Lomitapide.

[0111] In an embodiment, use of oral compositions generally comprises an inert diluent or an edible carrier. For the purpose of oral therapeutic administration, the active compound can be incorporated with excipients and used in the form of tablets, troches, or capsules, e.g., gelatin capsules. Pharmaceutically compatible binding agents, and / or adjuvant materials can be included as part of the composition. The tablets, pills, capsules, troches and the like can contain any of the following ingredients, or compounds of a similar nature: a binder such as microcrystalline cellulose, gum tragacanth or gelatin; an excipient such as starch or lactose, a disintegrating agent such as alginic acid, Primogel, or corn starch; a lubricant such as magnesium stearate or Sterotes; a glidant such as colloidal silicon dioxide; a sweetening agent such as sucrose or saccharin; or a flavoring agent such as peppermint, methyl salicylate, or orange flavoring. Formulations for oral use may be in the form of tablets which may be obtained by mixing active ingredients with known excipients. The tablet may also comprise of bilayer or film or gelatin coated by coating cores produced analogously to the tablets with substances normally used for coating. To achieve immediate release, the tablet may contain a suitable excipient, as herein described, along with the active ingredients.

[0112] In another embodiment, suitable tablets may be obtained for example, by mixing at least one of the compounds that may be used in the present invention with known excipients, for example diluents such as microcrystalline cellulose, calcium carbonate, calcium phosphate or lactose, disintegrants such as croscaramellose sodium, HPMC, sodium starch glycolate, binders such as starch or gelatine, guar gum, xanthum gum, lubricants such as magnesium stearate or talc and / or agents. The shapes include round, caplet, flat, oval and bevelled edges with and without embossing.

[0113] In another embodiment, the effective dose of the composition as disclosed in various embodiments herein may be administered as two, three, four, five, six or more sub-doses, administered separately at appropriate intervals throughout the day.

[0114] It is understood that various such modifications in the composition would be apparent to a person skilled in the art in light of the disclosures made herein and are included within the scope of the embodiments herein.

[0115] In another embodiment, a method of delaying primary tumor metastasis using the composition as disclosed herein. The method, according to embodiments herein includes administering to a subject in need thereof, a therapeutically effective amount of a composition including Lomitapide.

[0116] In one embodiment, the method includes administering to a subject in need thereof, a therapeutically effective amount of a composition including Lomitapide or pharmaceutically acceptable salt, ester, solvate thereof.

[0117] In one other embodiment, the method includes administering to a subject in need thereof, a therapeutically effective amount of a composition including Lomitapide either alone, or in combination with one or more therapeutic compound.

[0118] In one another embodiment, the invention is further described by reference to the following examples by way of illustration only and should not be construed to limit the scope of the aspects disclosed herein. It will be apparent to those skilled in the art that many modifications, both to materials and methods, may be practiced without departing from the scope of the claimed embodiments

[0119] WORKING EXAMPLES

[0120] The disclosure will now be illustrated with working examples, which is intended to illustrate the working of disclosure and not intended to take restrictively to imply any limitations on the scope of the present disclosure. 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 this disclosure belongs. Although methods and materials similar to or equivalent to those described herein can be used in the practice of the disclosed methods and formulations, the exemplary methods, devices, and materials are described herein. It is to be understood that this disclosure is not limited to methods, and experimental conditions described, as such methods and conditions may vary.

[0121] Extensive experimentation and analysis were conducted to identify non-oncology drugs that may be used for delaying primary tumor metastasis.

[0122] EXAMPLE 1:

[0123] The most weighted steps driving successful metastasis were identified using proprietary platform and once the rate-limiting steps were identified in silico modelling was performed using machine learning algorithms to screen the ability of approved drugs for non-oncology indications to bind to at least two of these targets simultaneously.

[0124] Top ten drugs were selected and, the drugs were further tested across the critical weighted steps. EMT and Invasion represented the initial dissemination step (IV), whereas tumor cell-induced platelet activation (TCIPA) and extravasation (EV) represented the ability to survive in the blood. A primary focus was given on the final colonization stage, as the targets were more related to these. These included a change in the plasticity via MET, the ability to colonize (CLN), the ability to crosstalk with other cells (EU), and the ability to adapt (APT, HYPX, GO / M, ROS, APGY, etc.) All percentage inhibition data (or lack of it) were tabulated and analyzed by the Board Institute -based open software Morpheus to generate a heat map. K-means clustering analysis revealed that Lomitapide was very different from the other sets of compounds, which could be grouped into three clusters (FIG. 1).

[0125] Lomitapide stood out as the only compound in its cluster. Lomitapide was found to be the most potent compound with anti-metastatic effects in the proprietary platform and had a unique mode of action.

[0126] EXAMPLE 2:

[0127] In their prior research, the inventors of the present invention had demonstrated significance of Plasticity ratio (PR) as an important marker for cellular plasticity. They also highlighted that cells exhibiting lower PR tend to display more epithelial characteristics compared to those with higher PR, which have more mesenchymal characteristics. Cells with hybrid characteristics have a PR of around 1 and are highly plastic. Cells with lower PR have higher proliferating properties but lower metastatic properties, whereas those with higher PR are more metastatic and have lesser proliferation prowess.

[0128] To understand the effect of Lomitapide on metastasis, HT29 cell line obtained from ATCC was treated with 1 pM Lomitapide. HT-29 is a human colorectal adenocarcinoma cell line with epithelial morphology. These cells have a very low PR and are reported to be highly epithelial and non-metastatic. A significant increase in PR was observed, which increased the cells' mesenchymal properties. Lomitapide treatment of HT29 cells increased the amount vimentin expression, thereby increasing population of both Ecad negative Vim positive cells and dual Ecad, Vimentin positive cells (FIG. 2A and 2B).

[0129] There was a 13.6-fold increase in PR, showcasing the shift of cells from epithelial to mesenchymal phenotype, explaining the reduction of primary tumor size in the mice models.

[0130] EXAMPLE 3:

[0131] It was known to Applicant that a mere increase in PR might not be enough for successful metastasis. If cells are shifted to a significantly higher PR, they might become so mesenchymal that their impact on adaptation properties will abrogate their colonization effects. To further understand whether Lomitapide’ s impact was limited to a mild to moderate increase in PR that would get saturated or whether Lomitapide could drive cells to very high PR, making them so mesenchymal in nature that colonization would be prohibited, MET assay was performed. Pre- metastatic colon cancer cell line SW-480 obtained from ATCC was treated with Lomitapide. Dot-plot analysis revealed that Lomitapide treatment increases the expression of Vimentin and decreases the expression of E-cadherin (FIG 3 A).

[0132] Lomitapide was observed to increase the population of Ecad negative Vim positive cells, thereby increasing the PR of SW480 cells almost twofold, shifting it further towards the mesenchymal axis (FIG. 3B).

[0133] EXAMPLE 4:

[0134] The initial dissemination of primary tumor cells is marked by successful migration, invasion, and trans-endothelial migration (TEM) through either lymph or blood endothelial cells. Trans-well assays were performed using both HUVEC and HLEC cells to understand the effect of Lomitapide on TEM. 1 pM Lomitapide completely inhibited trans-endothelial migration across HUVEC cells, and its impact was observed to be comparable to that of the internal control cytochalasin D (FIG. 4A). Similarly, extravasation (EV) measures the tumor cells' ability to survive in the blood and eventually move out to a secondary tissue. 1 pM Lomitapide inhibited extravasation, by almost 50%, and was found to be half as potent as the positive control cytochalasin D (FIG. 4B).

[0135] EXAMPLE 5:

[0136] Next, Lomitapide’s impact on mesenchymal cells' adaptability status was studied. These cells survive in secondary tissue by upregulating ROS, HIFla. A higher ROS promotes more HIFla formation, and since Lomitapide showed a moderate increase in ROS levels of tumor cells, the effect of Lomitapide on the HIFla levels of tumor cells were also studied.

[0137] SW480 cells treated with the HIFla inhibitor IDF-11774 showed a significant reduction in the positive percent population of HIFla cells. Conversely, Lomitapide, at a concentration of ImM, increases the percent population of HIFla -positive cells (FIG. 5A). Therefore, SW480 cells when treated with Lomitapide, showed a significant increase in the HIFla levels.

[0138] Higher ROS and hypoxia would maintain cells at the mesenchymal axis and also inhibit growth and re-colonization, therefore the effect of Lomitapide on colony formation assay was also studied. All-trans-retinoic acid (RA) was used as a positive control, as it has been reported to increase MET and, and consequently promote colonization. The known MET inhibitor AGN193109 was also used as an internal control. RA increased colonization of SW480 cells, which AGN193109 inhibited. AGNI 93109 converted cells more to their mesenchymal form and significantly reduced colony formation, whereas Lomitapide (ImM) prevented colony formation (FIG. 5B).

[0139] EXAMPLE 6:

[0140] The next step was to see if the higher mesenchymal nature of cells had any effect on senescence. For the study, instead of cell lines, primary cells from patients with primary colorectal cancer tumors were isolated and treated with Lomitapide. Interestingly, ImM Lomitapide increased senescence in the patient's primary tumor cells (CRC036 and CRC058), obtained from patients, Rajiv Gandhi cancer research Hospital diminishing their re-epithelialization chances (FIG. 6). Senescence of SW480 cells was selected as an internal control. EXAMPLE 7:

[0141] Next, the effect of Lomitapide on metastasis in Drosophila was studied. The larval progeny, i.e., the Fl generation for the experimentation, had the following genotype: vg Gal4; UAS-GFP>UAS-Scrib RNAi: UAS-RasV12. High dietary sugar (HDS) media was prepared using lOmM sucrose to promote metastasis. The larval progeny of the wild types, not treated with Lomitapide, showed significant tumor growth and metastasis. A significant increase of metastasis in the larval stage, moving from day 5 to day 8 was observed.

[0142] Lomitapide at a concentration of 10 mM was observed to inhibit metastasis on day 5 but not days 6 and 8. However, a higher concentration of 30 pM was observed to completely inhibit metastasis on days 5 and 6, and almost by 90% on day 8 (FIG. 7). The study was terminated on day 8, as the active larval population tends to change into a pupa. This in vivo fly data correlated with in vitro data of shifting cells to the mesenchymal axis and therefore preventing metastasis.

[0143] EXAMPLE 8:

[0144] Engineered HT29 cells (source-PCT / IN2022 / 050928) that were highly metastatic, HT29#12BC6, were subcutaneously implanted into NOD SCID mice. The tumor was allowed to grow for 14-20 days, following which the mice were divided into different treatment groups. The mice were randomized into these groups to ensure equal distribution. Two doses of Lomitapide (5 and 10 mg / kg) were administered orally to the treatment groups, while the control group received no treatment (only the vehicle used for Lomitapide formulation was administered). Each treatment group consisted of 8 animals, and dosing continued for 30 days. Lung and liver tissues were isolated and analyzed for signs of metastasis using H&E staining and digitization techniques. Artificial intelligence using the publicly available tools line algorithm https: / / qbrc.swmed.edu / projects / ai-tools was employed for tumor stroma analysis and quantifying lung and liver metastatic lesions. FIG. 8A depicts a flow chart of the animal model experimentation.

[0145] FIG. 8B depicts effect of 5 and 10 mg / kg Lomitapide on the primary tumor size. Statistically significant reduction of tumor volume was observed at lOmg / kg dose but not with 5 mg / kg dose. Further, both 5 and 10 mg / kg of Lomitapide was found to reduce the number of metastatic lesions in the animals' lungs (FIG. 8C) and liver (FIG. 8D).

[0146] The dose-response in the liver was significant compared to the dose response in the lung.

[0147] EXAMPLE 9:

[0148] A study was undertaken to measure the dose response and the effects of composition used in the present method on metastasis in a colorectal carcinoma ectopic model, using the HT29#12BC6 (source- PCT / IN2022 / 050928) cell line in female NOD SCID mice, with doses equivalent to clinical levels. The present composition used in the claimed method was administered orally at a q.d. regimen for 30 days, focusing on the relationship between drug exposure and anti-metastatic effects.

[0149] The tumor was allowed to grow for 14-20 days, following which the mice were divided into different treatment groups. The mice were randomized into these groups to ensure equal distribution. Three doses of Lomitapide (1, 3.5 and 12.3 mg / kg) were administered orally to the treatment groups, while the control group received no treatment (only the vehicle used for Lomitapide formulation was administered). The 12.3 mg / kg dose was equivalent to a human dosage of 60 mg per adult. Each treatment group consisted of 8 animals, and dosing continued for 30 days. Lung and liver tissues were isolated and analyzed for signs of metastasis using H&E staining and digitization techniques. Artificial intelligence using the publicly available algorithm https: / / qbrc.swmed.edu / projects / ai-tools was employed for tumor stroma analysis and quantifying lung and liver metastatic lesions.

[0150] Results- With respect to Primary Tumour Response:

[0151] The treatment by the present method using Lomitapide demonstrated a dosedependent reduction in tumor volume over 29 days. The highest dose (12.3 mg / kg) showed the most significant tumor suppression, achieving 77.17% tumor growth inhibition (TGI) compared to the vehicle control. Intermediate doses (3.5 mg / kg and 1 mg / kg) resulted in 50.33% and 32.52% TGI, respectively, highlighting a clear dose-response relationship (Fig 9 A). This reduction in tumor volume matched with lomitapide exposure in the tumor tissue (ng / ml) ranging from 13 ng / ml for 1 mg / kg to 49 ng / ml for 3.5 mg / kg to 178 ng / ml for 12.3 mg / kg of lomitapide (Fig 9B).

[0152] Lomitapide was well tolerated and had no adverse effects on the body weights of mice, as evident with 12.3 mg / kg Lomitapide showing no reduction in weight compared to the untreated control (Fig. 9 C)

[0153] EXAMPLE 10:

[0154] Lomitapide exhibited a pronounced and dose-dependent response on lung metastasis, with lesion reduction improving progressively with increasing drug concentration (Fig 10A). The highest dose (12.3 mg / kg) resulted in an exceptional 91.1% reduction in metastatic lesions, demonstrating near-complete inhibition of lung metastases. The mid dose (3.5 mg / kg) reduced lung lesions by 46.1%, while the lowest dose (1 mg / kg) had a more modest effect, reducing lesions by only 18.2%. The decrease in metastasis correlated with a clear dose -dependent increase in systemic exposure, with plasma concentrations escalating from 39.60 ng / mL at the lowest dose (1 mg / kg) to 243.80 ng / mL at the highest dose (12.3 mg / kg), reflecting effective drug absorption (Fig 10B). Liver metastasis showed a notable reduction across all tested doses, with the highest suppression (84.5%) observed even at the lowest dose (1 mg / kg), suggesting a strong impact on hepatic metastatic lesions. At 3.5 mg / kg, metastases were reduced by 44.4%, while the highest dose (12.3 mg / kg) resulted in a 58.8% reduction (Fig 10C). This pattern indicates that MS-AP-030 effectively targets liver metastases at lower doses, potentially due to the liver’s role in drug metabolism and first-pass clearance, which may affect local drug exposure. However, the effect was not strictly dose-dependent, likely due to complex hepatic drug processing mechanisms influencing tissue concentrations.

[0155] Results-

[0156] The claimed method using composition demonstrated strong anti-metastatic effects in a dose-dependent manner, particularly against lung metastases, where the highest dose (12.3 mg / kg) led to a 91.1% reduction in metastatic burden. Additionally, liver metastases were significantly reduced, particularly at the lowest dose (1 mg / kg), indicating a complex interplay between drug exposure and organ-specific metastasis suppression.

[0157] The findings suggest that the claimed method with composition exerts a dual therapeutic effect, inhibiting both primary tumor proliferation and metastatic spread.

[0158] The 12.3 mg / kg dose appears to be optimal, balancing efficacy with tolerability. Based on these results, the extrapolated human dose required to achieve comparable plasma exposure would be approximately 60 mg, supporting its potential for clinical development as an anti-metastatic agent in colorectal cancer.

[0159] The foregoing description of the specific embodiments will so fully reveal the general nature of the embodiments herein that others can, by applying current knowledge, readily modify and / or adapt for various applications such specific embodiments without departing from the generic concept, and, therefore, such adaptations and modifications should and are intended to be comprehended within the meaning and range of equivalents of the disclosed embodiments. It is to be understood that the phraseology or terminology employed herein is for the purpose of description and not of limitation. Therefore, while the embodiments herein have been described in terms of embodiments, those skilled in the art will recognize that the embodiments herein can be practiced with modification within the scope of the embodiments as described herein.

Claims

We Claim:

1. A method for delaying tumor metastasis, comprising: a. preparing a composition comprising Lomitapide and / or a pharmaceutically acceptable salt, ester, solvate, analogue, thereof or in combination with one or more therapeutic compound; and b. administering the therapeutic effective amount of the obtained composition in the range of 50-70 mg, to a subject in need thereof.

2. The method according to claim 1, wherein in the method using the composition further comprises a pharmaceutically acceptable carrier, excipient, diluent and / or adjuvant.

3. The method according to claim 1, wherein in the method using the composition, the pharmaceutical salts are selected from the group comprising mesylate, maleate, and camsylate.

4. The method according to claim 2, wherein in the method using the composition, the pharmaceutically acceptable carrier selected from the group comprising solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like, compatible with pharmaceutical administration.

5. The method according to claim 1, wherein in the method using the composition, the therapeutic compound is selected from the group comprising losartan, propranolol, metformin, aspirin, atenolol, niclosomide and combinations thereof.

6. The method according to claim 1, wherein in the method using the composition, the composition comprises Lomitapide and therapeutic compound in a ratio of 1:99.

7. The method according to claim 1, wherein the administering of the composition is in the form of tablets, troches, or capsules.

8. The method according to claim 1, wherein the method is used in the subject has been diagnosed from the group comprising colorectal cancer, head and neck cancers, triple-negative breast cancer, pancreatic cancer, liver cancer, ovarian cancer, esophageal cancer, cervical cancer, lung cancer, bladder cancer, and kidney cancer.

9. The method according to claim 1, wherein the method is used to administer the composition as two, three, four, five, six or more sub-doses, separately at appropriate intervals throughout the day.

10. The method according to claim 1, wherein the concentration of Lomitapide in the composition may range from as low as 0.1% of the total amount of the composition up to as high as 100%.

11. The method according to claim 1, wherein the dosage of the composition administered is in the range of 50-70 mg, to a subject in need thereof.

12. The method according to claim 1, wherein in the method using the composition, the composition administered comprises 50-65 mg of Lomitapide, pharmaceutically acceptable salts and therapeutic compounds.

13. The method according to claim 1, wherein in the method using the composition, the composition administered preferably comprises 60 mg of Lomitapide.

14. A composition of delaying tumor metastasis comprises Lomitapide and / or a pharmaceutically acceptable salt, ester, solvate, analogue, thereof and / or in combination with one or more therapeutic compound.

15. The composition according to claim 14, wherein the composition further comprises a pharmaceutically acceptable carrier, excipient, diluent and / or adjuvant.

16. The composition according to claim 14, wherein the therapeutic compound is selected from the group comprising losartan, propranolol, metformin, aspirin, atenolol, niclosomide and combinations thereof.

17. The composition according to claim 14, wherein the pharmaceutical salts are selected from the group comprising mesylate, maleate, and camsylate.

18. The composition according to claim 14, wherein the composition comprises lomitapide and therapeutic compound in a ratio of 1:99.

19. The composition according to claim 14, wherein pharmaceutically acceptable carrier selected from the group comprising solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like, compatible with pharmaceutical administration.

20. The composition according to claim 14, wherein the composition comprises 50-70 mg of Lomitapide, pharmaceutically acceptable salts and therapeutic compounds.

21. The composition according to claim 14, wherein the composition preferably comprises 60 mg of Lomitapide.

22. A composition for the treatment of delaying tumor metastasis comprising: Lomitapide and or a pharmaceutically acceptable salt, ester, solvate, analogue, thereof or in combination with one or more therapeutic compound in range of 50-70mg.

23. The composition according to claim 22, wherein the composition comprises 50-65 mg of Lomitapide, pharmaceutically acceptable salts and therapeutic compounds.

24. The composition according to claim 22, wherein the composition using Lomitapide for treating delaying tumor metastasis preferably comprises 60 mg of Lomitapide.

25. Use of composition for delaying tumor metastasis comprising: a) preparing a composition Lomitapide and or pharmaceutically acceptable salt or in combination with one or more therapeutic compound; and b) administering the therapeutic effective amount of the obtained composition in the range of 50-70 mg to a subject in need thereof.

26. The use of the composition according to claim 25, wherein the composition for delaying tumor metastasis comprises 50-65 mg of lomitapide, pharmaceutically acceptable salts and therapeutic compounds.

27. The use of the composition according to claim 25, wherein the composition for delaying tumor metastasis preferably comprises 60mg of lomitapide.

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