WO2025224756A1 - Composition, method and use thereof for delaying primary tumor metastasis using amlodipine - Google Patents

Abstract

The present disclosure provides a method for delaying preliminary tumor metastasis. The method involves: (a) preparing a composition containing at least one calcium channel blocker (CCB), or a pharmaceutically acceptable salt thereof, either alone or in combination with one or more therapeutic compounds; and (b) administering a therapeutically effective amount of the composition, typically in the range of 2-16 mg, to a subject in need. The disclosure particularly pertains to the use of such compositions for delaying tumor metastasis, with Amlodipine being the most preferred calcium channel blocker in the formulation.

Description

COMPOSITION, METHOD AND USE THEREOF FOR DELAYING PRIMARY TUMOR METASTASIS USING AMLODIPINE

TECHNICAL FIELD

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

BACKGROUND

Metastasis is a pivotal and intricate process in cancer progression where malignant cells spread from the primary tumor to other parts of the body via blood or lymphatic systems. This process involves several steps, such as detachment from the primary tumor, invasion into surrounding tissues, circulation through blood or lymph, and establishment of secondary tumors in new tissues. Cellular transitions like Epithelial mesenchymal transition (EMT) and mesenchymal epithelial transition (MET) play crucial roles in enhancing motility and facilitating colonization, respectively.

Metastasis is a major contributor to cancer-related mortality and poor prognosis, particularly in colorectal cancer, where liver and lung metastasis are common. Despite advances in treatment, effective therapies targeting metastasis remain limited, as many clinical trials targeting this process have faced challenges.

Metastasis is a hallmark of cancer progression, wherein malignant cells spread from their primary site to other parts of the body through the bloodstream or lymphatic system. This process poses a significant challenge in cancer treatment due to its complexity and the difficulty in eradicating metastatic lesions.

The delay in metastasis refers to the time period between the initial development of cancer in the primary site and the appearance of metastasis (spread of cancer to other parts of the body). This delay can vary depending on various factors such as the type of cancer, the biological characteristics of the tumor, and the environment of the body. Delays in treatment for metastatic cancer exceeding 12 weeks have been linked to adverse survival outcomes. Additionally, a 6-month delay in cancer treatment was associated with an estimated 21.3% increase in local tumor control risk and a 6.0% increase in distal metastasis risk.

Delaying tumor metastasis, or the spread of cancer cells from their primary site to other parts of the body, is a key focus in cancer research. Preventing or slowing this process can significantly improve prognosis and survival rates for patients with cancer. These findings emphasize the importance of minimizing delays in cancer treatment to improve survival rates and reduce the risk of metastasis.

Cancer cells undergo a series of intricate steps to metastasize, including detachment from the primary tumor, invasion through the surrounding epithelium, intravasation into blood or lymphatic vessels, survival in the circulation, extravasation at distant sites, and colonization in new tissues to establish secondary tumors. During metastasis, tumor cells may undergo EMT, a process that enhances their motility and invasiveness, allowing them to penetrate through basement membranes and invade adjacent tissues. Conversely, MET may occur at secondary sites, facilitating the colonization of distant organs by enabling cancer cells to establish proliferative colonies.

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. The present cancer therapies effectively confront most of the challenges: chemotherapy and radiotherapy manage proliferation, targeted therapy addresses initiation, while immunotherapy promotes the patient's immunity. However, there's a notable gap in treatments capable of delaying metastasis, which is synonymous with the terminal stage of cancer.

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.

Evidently, there is a significant gap that exists in comprehending and effectively treating metastasis, especially in establishing these treatments as primary options in adjuvant or neo-adjuvant settings. Furthermore, any such treatments must adhere to stringent safety standards, given that patients with primary tumors would necessitate chronic administration of these prophylactic metastasis treatments.

The complexity of metastasis underscores the urgent need for innovative therapeutic strategies to improve outcomes in cancer care.

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.

SUMMARY

In an aspect of the present disclosure, a method for delaying tumor metastasis, comprising: a. preparing a composition comprising at least one calcium channel blocker (CCBs) 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 2-16 mg, to a subject in need thereof.

In yet another aspect of the present disclosure, in the method using the composition, the calcium channel blocker is selected from the group comprising non- dihydropyridines and dihydropyridines.

In a further aspect of the present disclosure, in the method, at least one calcium channel blocker is a dihydropyridine calcium antagonist.

In another aspect of the present disclosure, in the method, at least one dihydropyridine calcium antagonist is Amlodipine.

In another aspect of the invention, in the method, the composition further comprises a pharmaceutically acceptable carrier, excipient, diluent and/or adjuvant.

In another aspect of the invention, in the method, the pharmaceutical salts are selected from the group comprising besylate, mesylate, maleate, and camsylate.

In another aspect of the invention, in the method, 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.

In another aspect of the invention, in the method, the therapeutic compound is selected from the group comprising losartan, propranolol, metformin, aspirin, atenolol, niclosomide and combinations thereof.

In another aspect of the invention, in the method, the composition comprises Amlodipine and therapeutic compound in a ratio of 1:99.

In another aspect of the invention, in the method, the composition is administered in the form of tablets, troches, or capsules. In another aspect of the invention, 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 invention, the composition is administered as two, three, four, five, six or more sub-doses, separately at appropriate intervals throughout the day.

In another aspect of the invention, in the method, using the composition the concentration of at least one calcium channel blocker in the composition may range from as low as 0.1% of the total amount of the composition up to as high as 100%.

In another aspect of the invention, in the method, the dosage of the composition is in the range of 2-16 mg per person per day.

In another aspect of the invention, in the method, the composition administered comprises 2-12 mg of amlodipine, pharmaceutically acceptable salts and therapeutic compounds.

In another aspect of the invention, the composition administered preferably comprises 10 mg of amlodipine.

In another aspect of the invention, a composition of delaying tumor metastasis comprises Amlodipine and pharmaceutically acceptable salt or in combination with one or more therapeutic compounds is provided.

In another aspect of the invention, the composition further comprises a pharmaceutically acceptable carrier, excipient, diluent and/or adjuvant.

In another aspect of the invention, the therapeutic compound is selected from the group comprising losartan, propranolol, metformin, aspirin, atenolol, niclosomide and combinations thereof. In another aspect of the invention, pharmaceutical salts are selected from the group comprising besylate, mesylate, maleate, and camsylate.

In another aspect of the invention, the composition comprises Amlodipine and therapeutic compound in a ratio of 1:99.

In another aspect of the invention, 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.

In another aspect of the invention, the composition comprises 2-12 mg of amlodipine, pharmaceutically acceptable salts and therapeutic compounds.

In another aspect of the invention, the composition preferably comprises 10 mg of amlodipine.

In another aspect of the invention, use of amlodipine for delaying tumor metastasis comprising: a composition comprising at least one calcium channel blocker (CCBs) and or pharmaceutically acceptable salt or in combination with one or more therapeutic compound; and the therapeutic effective amount of the obtained composition in the range of 2-16 mg is provided to a subject in need thereof.

In another aspect of the invention, use of amlodipine for delaying tumor metastasis comprising: 2-12 mg of amlodipine, pharmaceutically acceptable salts and therapeutic compounds.

In another aspect of the invention, use of amlodipine for delaying tumor metastasis preferably comprises 10 mg of amlodipine.

In some other aspect of the invention, the composition is a combination with Amlodipine and another therapeutic compound. In some other aspect of the invention, the composition comprises 2-12 mg of amlodipine, pharmaceutically acceptable salts and therapeutic compounds.

In another aspect of the invention, the amlodipine may inhibit tumor cell extravasation by at least about 50% compared to a control.

In another aspect of the invention, a method for inhibiting tumor cell intravasation in a subject with cancer is provided.

In another aspect of the invention, the method includes administering to the subject an effective amount of amlodipine.

In another aspect of the invention, the amlodipine may inhibit tumor cell intravasation by at least about 90% compared to a control.

In another aspect of the invention, a method for reducing platelet aggregation associated with colorectal cancer in a subject is provided. The method includes administering to the subject an effective amount of amlodipine.

In another aspect of the invention, the amlodipine may reduce platelet aggregation by at least about 50% compared to a control in present method.

In another aspect of the invention, the amlodipine may decrease E-cadherin expression in the colorectal cancer cells in present method. The amlodipine may increase vimentin expression in the colorectal cancer cells in the present method. In an exemplary aspect, Amlodipine partially inhibits the uptake of exosomes, extracellular vesicles that facilitate communication between tumor cells and their microenvironment. By disrupting exosome uptake, Amlodipine interferes with the crosstalk between tumor cells and the secondary tissue, delaying the adaptation of the tumor cells to the new environment and preventing the initiation of secondary metastasis.

In another aspect of the invention, use of amlodipine for the manufacture of a medicament for treating metastasis in a subject with colorectal cancer is provided. In another aspect of the invention, the medicament may be for oral administration. The medicament may be formulated for administration at a dose of about 2 mg to about 16 mg per day.

In another aspect of the invention, use of amlodipine for the manufacture of a medicament for inhibiting tumor cell extravasation in a subject with colorectal cancer is provided.

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.

BRIEF DESCRIPTION OF DRAWINGS

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:

FIG. 1A depicts a graph showing retrospective trial strategy, highlighting patient selection, timelines, and the data collected, in accordance with an aspect of the present disclosure;

FIG. IB depicts a graph showing validation of the Machine Learning (ML)-based optimum feature selection, in accordance with an aspect of the present disclosure;

FIG. 2 depicts a graph showing correlation between different drugs and patient survival using a machine learning model, in accordance with an aspect of the present disclosure;

FIG. 3 depicts a graph of a heat map showing hierarchical clustering of five pharmaceutical compounds as per their performance in phenotypic cellular assays, in accordance with an aspect of the present disclosure; FIG. 4A depicts a graph showing invasion assay using Amlodipine at a concentration of 2pM and Cytochalasin D; in accordance with an aspect of the present disclosure;

FIG. 4B depicts a graph showing EMT Assay, and the effect of it in E cadherin and Vimentin expression in accordance with an aspect of the present disclosure;

FIG. 4C depicts a graph showing change in PR (ratio of Vimentin to E-cadherin), in accordance with an aspect of the present disclosure;

FIG. 5A depicts a graph showing MET Assay with SW480 cells and Amlodipine, and the effect of it in E cadherin and Vimentin expression, RA acts as a positive control for MET, in accordance with an aspect of the present disclosure;

FIG. 5B depicts a graph showing increase in PR of SW480 cells by Amlodipine, in accordance with an aspect of the present disclosure;

FIG. 6A depicts a graph showing trans-endothelial cell migration assay of tumor cells across HUVEC cells, in accordance with an aspect of the present disclosure;

FIG. 6B depicts a graph showing Intravasation assay using tumor cells, HUVEC and Platelet-rich plasma (PRP), in accordance with an aspect of the present disclosure;

FIG. 7 A depicts a graph showing reduced tumor cell-induced platelet aggregation by 2pM Amlodipine, in accordance with an aspect of the present disclosure;

FIG. 7B depicts a graph showing inhibition of extravasation of the tumor cells by 2pM Amlodipine, in accordance with an aspect of the present disclosure;

FIG. 8A depicts a chart showing animal model experimentation, in accordance with an aspect of the present disclosure; FIG. 8B depicts a graph showing effect of 5 and 10 mpk (mg/kg) Amlodipine on the primary tumor size, in accordance with an aspect of the present disclosure;

FIG. 8C depicts a graph showing effect of 5 and 10 mpk Amlodipine on lung metastasis, in accordance with an aspect of the present disclosure;

FIG. 8D depicts a graph showing effect of 5 and 10 mpk Amlodipine on liver metastasis, in accordance with an aspect of the present disclosure;

FIG. 9A depicts a graph showing that amlodipine has no effect with increasing dosage of 1, 1.5 and 2 mpk on reducing the primary tumor volume, inset shows that the lack of efficacy is not due to the lack of bioavailability, as there is significant increase in plasma concentration of amlodipine, with increasing dosage,

FIG. 9B depicts a graph showing the lack of significant weight loss, even at the highest dose of 2 mg/kg, implies that amlodipine is well-tolerated, in accordance with an aspect of the present disclosure;

FIG. 10A depicts a graph showing the low dose of amlodipine (1 mg/kg) led to a 76.5% reduction in liver lesions (16 lesions), while the high dose (2 mg/kg) caused a 55.9% decrease (30 lesions), in accordance with an aspect of the present disclosure;

FIG. 10B depicts a graph showing the substantial 58.3% reduction by mid dose, whereas the low dose showed no significant impact on lung metastasis, and the high dose showing 88.3 % reduction, aligning with the PK/PD correlation, in accordance with an aspect of the present disclosure;

FIG. 11A depicts a graph showing the bioavailability study of amlodipine, with IV dose of 2mg/kg (11 A) , dosed in 3 mice for each group, in accordance with an aspect of the present disclosure; FIG. 11 B depicts a graph showing the PK study of Amlodipine after PO Administration (10 mg/kg dose), dosed in 3 mice for each group, in accordance with an aspect of the present disclosure;

FIG 11 C depicts a table showing the data correlated with historical values, with amlodipine showing high bioavailability and volume of distribution, in accordance with an aspect of the present disclosure;

FIG. 12 A depicts a graph showing the administration amlodipine (2 mg/kg) did not lead to significant changes in body weight compared to the Vehicle Control group during the observation period, in accordance with an aspect of the present disclosure;

FIG. 12 B depicts a graph showing unlike the positive control carrageenan, which showed significant paw edema, amlodipine had no effect and was comparable to control, as on day 1, in accordance with an aspect of the present disclosure; and

FIG. 12 C depicts a graph showing the course of treatment of 28 days, amlodipine showed no contribution towards either erythema or edema, was comparable to the control, in accordance with an aspect of the present disclosure.

To facilitate understanding, reference numerals have been used, where possible, to designate elements common to the figures.

DETAILED DESCRIPTION

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. 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.

The words/phrases "exemplary", “example”, “illustration”, “in an instance”, “and the like”, “and so on”, “etc.”, “etcetera”, “e.g.,”, “i.e.,” 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”, “e.g.,”, “i.e.,” is not necessarily to be construed as preferred or advantageous over other embodiments.

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.

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.

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.

The term “Calcium channel blocker” or “Calcium channel antagonists” as used interchangeably herein, refers to heterogenous group of drugs that prevent or slow the entry of calcium into cells by regulating cellular calcium channels. These drugs are generally known to be used for treating cardiovascular disease, pulmonary hypertension, peripheral vascular disorder, migraine disorder, mania, epilepsy, depression, hyperuricemia, asthma, Raynaud phenomenon, subarachnoid hemorrhage, migraine headaches, and so on.

Calcium channel blocker is further classified as non-dihydropyridines and dihydropyridines. In one embodiment, at least one calcium channel blocker is a dihydropyridine calcium antagonist. Non-limiting examples of dihydropyridine calcium antagonist include Nifedipine, Amlodipine, Felodipine, Nicardipine, Isradipine, Nimodipine, Lacidipine, Lercanidipine, Nitrendipine, Clevidipine, Benidipine, Azelnidipine, Barnidipine, Cilnidipine, Manidipine, Efonidipine, Darodipine and so on. In one embodiment, at least one calcium channel blocker is Amlodipine. In one embodiment, at least one calcium channel blocker is Amlodipine or pharmaceutically acceptable salt, ester, solvate, analogue, thereof.

Calcium channel blockers that is used in this invention is Amlodipine (It is marketed under the trade names Norvasc (Pfizer), Istin (Pfizer), Amlostin (Sun Pharmaceutical Industries Ltd), Amlod (Sandoz (Novartis)), Stamlo (Dr. Reddy's Laboratories), Amcard (Torrent Pharmaceuticals Ltd), Vamlo (Alembic Pharmaceuticals Ltd), Amlopres (Cipla Ltd.), Amdepin (Zydus Cadila), and Amlong (Micro Labs Ltd), to name a few.

It is well known in prior art that Amlodipine is primarily used as a medication for the treatment of hypertension and angina. Amlodipine acts as a peripheral arterial vasodilator by directly diminishing peripheral vascular resistance, thereby reducing blood pressure.

The present invention seeks to address a longstanding gap in cancer research, where previous studies have predominantly focused on the inhibition of tumor cell migration and invasion, key stages of cancer progression. These investigations have relied on extensive compound screening to identify agents capable of suppressing one or both processes. While such approaches have yielded significant insights, they have primarily targeted a limited aspect of cancer metastasis. The majority of existing strategies aimed at controlling cancer metastasis have concentrated on halting the ability of cancer cells to migrate from the primary tumor or invade neighboring tissues. Compounds identified through these studies were designed to inhibit cellular motility or degrade pathways critical for tissue invasion. Despite these advancements, such methods fail to account for the complexity of subsequent steps in the metastatic cascade, such as intravasation, survival in circulation, extravasation, and colonization in secondary tissues. Moreover, many previously identified compounds exhibit off-target effects or cytotoxicity, restricting their therapeutic applicability. Furthermore, these methods do not address the dynamic plasticity of tumor cells that enables them to adapt and evade treatment over time. The five identified compounds were further tested across a proprietary panel, particularly the weighted steps. All the five identified compounds were tested at non-cytotoxic concentrations that would not adversely impact cell viability. This is important, so as to differentiate anti-metastatic activity from anti-proliferation effects. The identified five compounds were tested on epithelial-to-mesenchymal transition (EMT), invasion, trans-endothelial migration, intravasation, platelet binding, extravasation, apoptosis, exosomal crosstalk and mesenchymal-to- epithelial transition (MET).

Proprietary platforms and algorithms have identified non-oncology drugs with the potential to hinder metastasis. Remarkably, Amlodipine, a drug primarily known for its use as a calcium channel blocker in cardiovascular treatments, was found to possess unexpected effects in the context of tumor cell behavior and metastasis.

Amlodipine influences tumor cell plasticity by shifting the cells towards a mesenchymal phenotype. This is reflected in an increase in the Plasticity Ratio (PR), a key indicator of the ability of tumor cells to undergo epithelial-to- mesenchymal transition (EMT) and contribute to metastasis. The increase in PR, resulting from Amlodipine treatment, theoretically leads to enhanced metastatic potential. Amlodipine-treated cells demonstrate characteristics of stem-like cells and lose their ability to revert back to an epithelial form, a process central to the development of metastasis. Concurrent with the PR increase, there is a marked rise in the invasion of tumor cells. The drugs, pharmaceutically acceptable excipients, therapeutic compounds were all commercially procured and used in the present invention.

Amlodipine partially impedes the binding of tumor cells with platelets, an interaction that is known to be critical for the survival of circulating tumor cells. By reducing platelet-tumor cell binding, Amlodipine further decreases inhibition of extravasation, the survival chances of tumor cells in the blood, hindering their ability to thrive and establish secondary tumors.

Amlodipine, while facilitating the dissemination of tumor cells, does so in a manner that ultimately prolongs survival. The dissemination of cells from the primary tumor is expedited, but the cells are unable to establish secondary tumors due to the inability to fully transition back to the epithelial form and colonize new tissue. In the secondary tissue, Amlodipine does not promote mesenchymal-to-epithelial transition (MET), further reducing the likelihood of secondary tumor formation. Rather, Amlodipine reinforces the mesenchymal phenotype, maintaining the cells in their mesenchymal axis and preventing secondary tumor colonization.

The Applicant employed a Drosophila animal model to study the effects of Amlodipine on metastasis. The results demonstrated that Amlodipine significantly reduced metastasis without affecting the size of the primary tumor. This finding underscores the compound's ability to limit metastatic spread without altering the growth dynamics of the primary tumor, offering a potential therapeutic advantage in controlling secondary tumor formation without compromising the treatment of the primary malignancy.

During the animal studies it is observed that Amlodipine significantly reduced metastasis in the liver and lung without influencing the growth of the primary tumor. These findings further support the potential of Amlodipine to mitigate the spread of tumors to distant organs, specifically the liver and lung, which are common sites of metastasis, without affecting primary tumor progression. The pharmacokinetic properties of Amlodipine were also examined in relation to its effectiveness in reducing secondary tumor colonization. Amlodipine has a high volume of distribution (21 L/kg) and a mean plasma half-life of approximately 35 hours in humans, ensuring extensive tissue distribution and bioavailability. These characteristics are crucial for achieving sufficient drug concentration in tissues to effectively delay secondary tumor colonization, even in distant organs.

Further the Applicant firstly observed and conducted experiments with regard to Amlodipine- as a drug that can delay primary tumor metastasis by facilitating the dissemination of tumor cells without allowing them to establish secondary tumors. Amlodipine achieves this by increasing the PR, reinforcing the mesenchymal phenotype of tumor cells, inhibiting platelet-tumor cell binding, and partially blocking exosome uptake. These mechanisms together offer a novel therapeutic strategy for prolonging survival in cancer patients by limiting metastatic spread while not inhibiting the growth of the primary tumor.

In an embodiment, the present invention provides a novel approach utilizing a nononcology drug, Amlodipine, to delay primary tumor metastasis. After extensive research and analysis, it is discovered that Amlodipine exhibits unique properties that influence tumor cell behavior and metastatic progression. Amlodipine promotes tumor cell invasion by shifting the cellular phenotype toward the mesenchymal axis. Through rigorous experimentation, it has been demonstrated that Amlodipine treatment leads to a significant increase in the Plasticity Ratio (PR), which represents the ability of cells to adopt mesenchymal characteristics. Elevated PR levels are theoretically associated with enhanced metastatic potential. Amlodipine-treated cells exhibit increased stem-like properties and a diminished capacity to revert to their epithelial form, which further reinforces the mesenchymal phenotype. This sustained mesenchymal state promotes cellular dissemination from the primary tumor. While Amlodipine enhances tumor cell invasion, it concurrently inhibits critical steps in metastasis, including intravasation (the entry of tumor cells into the bloodstream) and extravasation (the exit of tumor cells from the bloodstream into secondary tissues). The reduction in these processes decreases the overall number of circulating tumor cells (CTCs), thereby mitigating the ability of the disseminated cells to colonize distant sites. Additionally, Amlodipine partially disrupts the binding interaction between tumor cells and platelets. This interference compromises the survival chances of CTCs in the bloodstream, further restricting metastatic progression.

In secondary tissues, Amlodipine impedes mesenchymal-to-epithelial transition (MET), a critical step for establishing metastatic colonies. Instead, Amlodipine reinforces the mesenchymal phenotype in disseminated tumor cells, maintaining a high PR. This increased PR prevents the colonization of secondary sites by suppressing the adaptation required for tumor cell survival and growth in the new microenvironment.

Amlodipine has been found to partially inhibit exosome uptake in secondary tissues, thereby reducing intercellular communication and crosstalk within the metastatic niche. This effect delays the adaptation of disseminated cells to the secondary microenvironment, offering additional protection against secondary tumor growth.

Amlodipine provides a novel therapeutic strategy to extend patient survival by delaying primary tumor metastasis. While Amlodipine enhances tumor cell dissemination, it effectively prevents these cells from successfully forming secondary tumors. The dual action of promoting invasion while inhibiting intravasation, extravasation, and secondary colonization represents a significant advancement in the management of metastatic cancer.

In an embodiment, the present invention provides a method for delaying tumor metastasis, comprising: a. preparing a composition comprising at least one calcium channel blocker (CCBs) 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 2-16 mg, to an adult subject in need thereof.

In yet another embodiment, in the method the calcium channel blocker is selected from the group comprising non-dihydropyridines and dihydropyridines.

In further another embodiment, in the method the at least one calcium channel blocker is a dihydropyridine calcium antagonist.

In another embodiment, in the method the at least one dihydropyridine calcium antagonist is Amlodipine.

In another embodiment, in the method the composition further comprises a pharmaceutically acceptable carrier, excipient, diluent and/or adjuvant.

In another embodiment, in the method the pharmaceutical salts are selected from the group comprising besylate, mesylate, maleate, and camsylate.

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

In another embodiment, in the method the composition comprises Amlodipine and therapeutic compound in a ratio of 1:99.

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

In another embodiment, the method is used as 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 invention, in the method, the composition is administered as two, three, four, five, six or more sub-doses, separately at appropriate intervals throughout the day.

In another embodiment, in the method, the concentration of at least one calcium channel blocker in the composition may range from as low as 0.1% of the total amount of the composition up to as high as 100%.

In another embodiment, in the method, the dosage of the composition is administered in the range of 2-16 mg per person per day.

In another embodiment, in the method, the composition administered comprises 2- 12 mg of amlodipine, pharmaceutically acceptable salts and therapeutic compounds.

In another embodiment, in the method, the composition administered preferably comprises 10 mg of amlodipine. In one embodiment, the concentration of at least one calcium channel blocker in the composition is from 1% to 90% by weight.

In one other embodiment the concentration of at least one calcium channel blocker in the composition is from 5% to 80% by weight.

In one other embodiment the concentration of at least one calcium channel blocker in the composition is from 10% to 70% by weight.

In another embodiment, the dosage of composition that may be used 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 will be an amount which is the lowest dose effective to produce the desired effect with no or minimal side effects.

In another embodiment, the present disclosure provides a composition of delaying tumor metastasis comprises Amlodipine and pharmaceutically acceptable salt or in combination with one or more therapeutic compounds.

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

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

In another embodiment, the pharmaceutical salts are selected from the group comprising besylate, mesylate, maleate, and camsylate.

In another embodiment, the composition comprises Amlodipine and therapeutic compound in a ratio of 1:99. In another embodiment, 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.

In another embodiment, the composition comprises 2-12 mg of amlodipine, pharmaceutically acceptable salts and therapeutic compounds.

In another embodiment, the composition preferably comprises 10 mg of amlodipine.

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

In one embodiment, the composition includes a combination of Amlodipine and therapeutic compound.

In an embodiment, the method includes administering to a subject in need thereof, a therapeutically effective amount of a composition including at least one calcium channel blocker (CCBs).

In one other embodiment, the method includes administering to a subject in need thereof, a therapeutically effective amount of a composition including at least one calcium channel blocker either alone, or in combination with one or more therapeutic compound.

In an embodiment, the method may also include strategies to enhance the delivery or efficacy of amlodipine. In some cases, this may involve using drug delivery systems such as nanoparticles or liposomes to encapsulate amlodipine. These delivery systems may improve the pharmacokinetics and biodistribution of amlodipine, potentially enhancing its anti-metastatic effects. In an embodiment, the method may include steps to manage or mitigate potential side effects associated with amlodipine treatment. This may involve administering supportive medications, adjusting the dosage, or implementing other strategies to improve tolerability and adherence to the treatment regimen.

In an embodiment, these delivery systems may improve the pharmacokinetics and biodistribution of amlodipine, potentially enhancing its anti-metastatic effects.

In an embodiment, the method may also include monitoring the subject's response to Amlodipine treatment. This monitoring may involve assessing tumor size, metastatic burden, biomarker levels, or other clinical parameters. In some cases, the method may include adjusting the Amlodipine dosage or treatment regimen based on the subject's response.

In certain embodiments, the method may involve administering Amlodipine via different routes of administration. While oral administration may be common, alternative routes such as intravenous injection, subcutaneous injection, or local delivery to the colorectal region may be employed in some cases. The route of administration may be selected based on factors such as the patient's condition, the stage of cancer, and the desired pharmacokinetic profile.

In an embodiment, the method may also include administering Amlodipine at different dosages or dosing schedules. In some cases, Amlodipine may be administered at a dose ranging from about 2 mg to about 12 mg per day. The dosing frequency may vary, and may include once daily, twice daily, or other dosing regimens as determined by a healthcare professional.

In some embodiments, the method may involve using Amlodipine in combination with other treatment modalities. For example, Amlodipine may be administered in conjunction with radiation therapy, surgical interventions, or other non- pharmacological treatments for colorectal cancer. In an embodiment, the method for treating metastasis in a subject with colorectal cancer may involve administering Amlodipine in combination with other therapeutic agents. The additional therapeutic agents may include chemotherapeutic drugs, targeted therapies, immunotherapies, or other anti-cancer treatments. For example, Amlodipine may be administered in conjunction with 5- fluorouracil, oxaliplatin, irinotecan, bevacizumab, cetuximab, or pembrolizumab.

In another embodiment, a composition of delaying tumor metastasis comprises Amlodipine and pharmaceutically acceptable salt or in combination with one or more therapeutic compounds.

In another embodiment, the composition further comprises a pharmaceutically acceptable carrier, excipient, diluent and/or adjuvant.

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

In another embodiment, the pharmaceutical salts are selected from the group comprising mesylate, maleate, and camsylate.

In another embodiment, the composition comprises Amlodipine and therapeutic compound in a ratio of 1:99.

In another embodiment, 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.

In another embodiment, the composition comprises 2-12 mg of amlodipine, pharmaceutically acceptable salts and therapeutic compounds.

In another embodiment, the composition preferably comprises 10 mg of amlodipine. In another embodiment, use of Amlodipine for delaying tumor metastasis comprising: a. preparing a composition comprising at least one calcium channel blocker (CCBs) and or pharmaceutically acceptable salt or in combination with one or more therapeutic compound; and b. the therapeutic effective amount of the obtained composition provided in the range of 2-16 mg to a subject in need thereof.

In another embodiment, use of the amlodipine for delaying tumor metastasis comprises 2-12 mg of amlodipine, pharmaceutically acceptable salts and therapeutic compounds.

In another embodiment, use of the amlodipine for delaying tumor metastasis preferably comprises 10 mg of amlodipine.

In an embodiment, use of oral compositions generally comprises an inert diluent or an edible carrier. For the purpose of oral therapeutic, 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 com 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. In one another embodiment, suitable tablets may be obtained for example, by mixing at least one of the compounds that may be used 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.

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.

WORKING EXAMPLES

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.

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

EXAMPLE 1:

Non-oncology drugs approved for chronic indications on the progression and survival of primary patient tumors was studied. A retrospective clinical trial (FIG. 1 A) was performed in collaboration with Rajiv Gandhi Cancer Centre and Research Institute, Rohini, New Delhi. Hundred colorectal cancer patients who were diagnosed with primary tumors and no clinical metastasis were selected for the study. They were treatment-naive, and all underwent surgery, followed by adjuvant chemotherapy. Follow-up was done, for a minimum of five years, on the patient's survival or progression. All clinical annotations, treatment, and a list of medications were recorded. These datasets were then pre-processed, and drug information was extracted for each patient, following which ML-based optimal feature selection was performed.

Next, Machine Learning (ML) classification model was trained and validated for patient survivability prediction and statistical tests were applied to correlate drugs with patient survival. Random forest classifier model has been used for both the determination of top features (Fig IB) and model prediction (Fig 2). This helped in identifying the top drugs having significant positive and negative correlations with survival.

Validation of the ML-based optimum feature selection showed parameters having a negative correlation with survival (FIG. IB). Serum CEA, produced by 90% of colorectal cancers, contributes to the malignant characteristics of a tumor and its plasma levels are used as a marker for prognosis. Serum CEA (black rectangle), an established marker for poor prognosis of colorectal cancer, was observed to negatively correlate with survival with a weightage of 0.2 (FIG. IB). Other parameters showed a higher weightage in their negative correlation with survival, with recurrence and progression having the maximum negative effect on survival, followed by metastasis and node status. This data validates the algorithm for its ability to select the optimal features.

Further, the identified drugs were correlated with patient survival using the same ML model. Amlodipine showed the maximum negative correlation with death, with a p-value <0.0001, followed by propranolol, losartan, metformin, and aspirin (FIG. 2). The p-value of aspirin was statistically insignificant. EXAMPLE 2:

Non-oncology drugs approved for chronic indications on the progression and survival of primary patient tumors was studied. The clinical trial database suggested propranolol, an anti-hypertension drug, that is widely being tested in adjuvant and neo-adjuvant settings to delay metastasis, with more than 50 such studies reported or ongoing, thereby validating our findings. Similarly, more than hundreds of ongoing studies on metformin, combined with other standard treatments, are reported in the clinical trial database. Aspirin has also been reported to be used in several cancer trials, combined with standard of care. Interestingly, there are few reports of Losartan and no reports of Amlodipine.

Hierarchical clustering suggested that Amlodipine was the most potent compound to have anti-metastatic effects in the proprietary panel, followed by propranolol, losartan, metformin and aspirin (FIG. 3). Interestingly, the rank order of these compounds in delaying or abrogating metastasis exactly matched the rank order of the compounds in retrospective studies, prolonging survival, thereby suggesting that survival prolongation was probably due to a delay in clinical metastasis.

EXAMPLE 3:

Non-oncology drugs approved for chronic indications on the progression and survival of primary patient tumors was studied.

Surprisingly, it was observed that amlodipine promoted the invasion of tumor cells. Invasion is a crucial aspect of metastasis and is synonymous with higher metastasis risk, per popular understanding. However, the proprietary platform identified successful re-epithelization or colonization as one of the other ratelimiting steps contributing to successful metastasis. As this colonization is dependent on the plasticity of the cells, possible role of Amlodipine in altering cellular plasticity was studied. HT29 cells, obtained from ATCC, when treated with EMT -promoting agent TGFp, showed an increase in the Vimentin population and a decrease in the E cadherin population, suggesting a change in the plasticity ratio (increasing PR), thereby moving cells towards the mesenchymal axis. Amlodipine was equally effective in promoting such EMT and increased the plasticity ratio. When added in combination with TGFp, Amlodipine shifted the cells completely to the mesenchymal axis. This effect of amlodipine, moving cells to the mesenchymal axis, explains the increased invasiveness of the cells but also suggests a probable low re-epithelization effect. Amlodipine was observed to increase invasion at a concentration of 2pM. Comparatively, the positive control cytochalasin D completely inhibited invasion (FIG. 4A). Amlodipine was equally effective as TGFP in promoting EMT and increased the vimentin population, decreasing the E-cadherin population (FIG. 4B). The effect of E-cadherin and Vimentin is summed up by the change in PR (ratio of Vimentin to E-cadherin). Amlodipine was observed to increase PR more than TGFp (FIG. 4C).

EXAMPLE 4:

Non-oncology drugs approved for chronic indications on the progression and survival of primary patient tumors was studied.

Now that it was established that Amlodipine increased Plasticity Ratio (PR), experiments were conducted to check if Amlodipine affected cells with higher PR, i.e., tumor cells already in the mesenchymal axis. The effect of all-trans-retinoic acid (RA) was tested on SW480 cells, obtained from ATCC, and post-treatment, these cells showed a mesenchymal to epithelial transition, with an increase in the E-cadherin population and a decrease in Vimentin population, consequently decreasing PR, shifting cells to the epithelial axis. Amlodipine, however, showed no such effect; instead, as was seen before, it shifted cells more towards the mesenchymal axis, increasing PR. This was conclusive evidence that Amlodipine shifts cells towards the mesenchymal axis and maintains them in that state. MET assay was performed with SW480 cells and Amlodipine. RA was selected as a positive control for MET. Amlodipine did not induce any MET but shifted cells more towards the mesenchymal axis, increasing PR (FIG. 5A). However, it could not compete with RA and prevent RA-induced MET. A three-fold increase in PR by Amlodipine was observed, shifting it towards the mesenchymal axis (FIG. 5B). EXAMPLE 5:

Non-oncology drugs approved for chronic indications on the progression and survival of primary patient tumors was studied. Next, the impact of Amlodipine, if any, on the trans-endothelial migration and intravasation of tumor cells was studied. Experiments were carried out as described previously. Conceptually, both are transwell migration, with the trans-endothelial cell migration assay using a HUVEC layer and FBS as a chemoattractant and Intravasation using HUVEC and Plateletrich plasma (PRP), isolated from patients or healthy donors as chemoattractant. 2pM of Amlodipine was observed to inhibit trans-endothelial migration across HUVEC cells completely and was comparable to the effect of cytochalasin D (FIG. 6A). Further, 2pM of Amlodipine almost completely inhibited intravasation through HUVEC against a PRP gradient and was comparable to the effect of the positive control, cytochalasin D (FIG. 6B).

EXAMPLE 6:

Non-oncology drugs approved for chronic indications on the progression and survival of primary patient tumors was studied. Further, the impact of amlodipine on cellular survival in the blood and extravasation abilities was studied. It is well known that platelet and tumor cell binding promote the survival and extravasation of tumor cells. Therefore, Amlodipine’s ability to abolish this interaction was studied. The data suggested that Amlodipine reduces platelet-induced tumor cell binding and activation by almost 50%, thereby reducing the chances of survival of tumor cells in the blood. Consequently, Amlodipine also inhibited extravasation of tumor cells from the vasculature.

SW480 cells were observed to bind to platelet-rich plasma (PRP) and promote platelet activation, as seen by the change in OD. 2pM Amlodipine partly inhibited this interaction (FIG. 7A). Similarly, 2pM Amlodipine almost wholly inhibited the extravasation of the tumor cells (FIG. 7B). EXAMPLE 7:

Non-oncology drugs approved for chronic indications on the progression and survival of primary patient tumors was studied. With all the in vitro data suggesting that amlodipine reduced secondary colonization without impacting the proliferation of the primary cells, the antimetastatic effect of Amlodipine in an animal model was studied. The animal model experiment was carried out using female NOD SCID mice and engineered cell lines HT29#12BC6 (FIG. 8A). In summary, engineered colon cancer lines (more metastatic) were injected subcutaneously in immune-compromised mice, tumors were allowed to grow for 20 days, and then animals were randomized into three groups, each having eight animals. One group was left untreated, whereas two groups were treated with 5 and 10 mpk (mg/kg) of Amlodipine. Oral dosing was done for four weeks, after which animals were sacrificed, their primary tumor volume was measured, and their lungs and liver were collected for H&E staining, followed by complete digitalization. The digital images were then assessed by Al-based software to identify tumor nuclei stroma in each lung or liver tissue, and the total area of these nuclei was measured by Image J analysis. Amlodipine had no effect on the size of the primary tumor but significantly decreased metastatic lesions in the lung and liver. The lower dose of 5mpk was more efficient in reducing the metastatic load.

FIG. 8B depicts effect of 5 and 10 mpk Amlodipine on the primary tumor size. No reduction of tumor volume was observed. 5mpk Amlodipine was shown to significantly reduce the number of metastatic lesions in the lung and the size of the metastatic nuclei (FIG. 8C). lOmpk Amlodipine was also shown to significantly reduce the size of the metastatic nuclei in the liver (FIG. 8D).

In vitro, at 2pm, assuming 100% bioavailability, amlodipine will be around 5.7 mg. Assuming 5 litres of blood in the human body, this is the rationale behind the 10 mg final dosage. Also 10 mg final dosage in human is equal to 2mg/kg final dose in mice (last experiment, PK/PD). All previous experiments were done at higher concentration in mice (5 and 10 mg/kg) as the Applicant wanted to show efficacy. But for dose correlation with human the Applicant reduced the dose in mice to 1, 1.5, and 2 mg/kg in the last experiment (lOmpk = 10 Mg Per Kg)

EXAMPLE 8:

Non-oncology drugs approved for chronic indications on the progression and survival of primary patient tumors was studied. Further, the impact of PK-PD correlation experiment of Amlodipine. Amlodipine showed no effect on primary tumour growth. By day 28, all treatment groups (G3-G5) displayed similar tumour volumes (-1100 mm³). Tumour growth was more pronounced in the normal feed group, suggesting that the TET diet may enhance the PR while inversely affecting tumour growth and potentially increasing metastasis in TET-fed animals (Fig 9A). This was despite significant increase in plasma concentration of amlodipine, with increasing dosage with 2mg/kg showing an average concentration of 93ng/ml amlodipine (Inset, Fig. 9A).

Body weight across all groups remained largely unchanged, indicating that Amlodipine is minimally toxic. The Vehicle Control (Gl) and Normal Feed (G2) groups exhibited no noteworthy weight changes, reflecting a stable health condition. Participants receiving Amlodipine at doses of 1 mg/kg (G3) and 2 mg/kg (G5) also showed consistent weight patterns, with only slight variations. Although the 1.5 mg/kg dose (G4) experienced a minor initial weight drop, it returned to stability by the end of the study. The lack of significant weight loss, even at the highest dose of 2 mg/kg, implies that amlodipine is well-tolerated (Fig. 9B).

EXAMPLE 9:

Non-oncology drugs approved for chronic indications on the progression and survival of primary patient tumors was studied. Further, the histopathological analysis of amlodipine treated liver and lung tissues showed significant decrease in lesions, compared to the untreated control. In the liver, the vehicle control group on the TET diet exhibited 68 metastatic lesions, reflecting a significant metastatic burden. The low dose of amlodipine (1 mg/kg) led to a 76.5% reduction in liver lesions (16 lesions), while the high dose (2 mg/kg) resulted in a 55.9% decrease (30 lesions) (Fig. 10A).

In terms of lung effects, the high dose proved particularly effective, decreasing lesions by 88.3% (7 lesions) compared to the 60 lesions in the vehicle group. The mid-dose also exhibited a significant reduction of 58.3%, while the low dose had no notable effect on lung metastasis, indicating a complete PK/PD correlation (Fig. 10B).

EXAMPLE 10:

Non-oncology drugs approved for chronic indications on the progression and survival of primary patient tumors was studied. Further, the impact of the bioavailability study of amlodipine, with IV dose of 2mg/kg and a PO dose of 10 mg/kg, respectively, dosed in 3 mice for each group (Fig. 11 A & 11B). The data correlated with historical values, with amlodipine showing high bioavailability and volume of distribution (Fig. 11C). The observed maximum dose exposure in the 2 mg/kg group was 93 ng/ml with a standard deviation of 22 ng/ml (inset, Fig. 9A) was comparable to the projected dose derived from the bioavailability study (Fig 11 C) as explained below-

93 ng/ml is the plasma concentration of amlodipine in the PKPD study (Inset, Fig 9 A) in the 2mg/kg dosage. As per the oral PK of 10 mg mean Cmax is 375.2 ng/ml (fig 11 C) which is five times the Cmax observed in the 2mg/kg dosage and comparable with the 93+/-22 ng/ml of amlodipine that was observed in this study.

EXAMPLE 11:

To evaluate hypotensive adverse effects, if any, of Amlodipine:

Non-oncology drugs approved for chronic indications on the progression and survival of primary patient tumors was studied. Further, the study evaluated the safety of amlodipine concerning erythema and edema in a preclinical model of BALB/c mice. It involved three groups: a vehicle control group (Gl), a carrageenan-induced inflammation positive control group (G2), and amlodipine treatment group (G3). In G2, inflammation was triggered by a single subcutaneous injection of 1% carrageenan into the paw on day 1. Amlodipine was orally given to G3 at a dose of 2 mg/kg once daily (q.d., p.o.) for 28 days. The vehicle control group (Gl) received the vehicle formulation of amlodipine, also dosed orally once daily for 28 days. The administration of amlodipine (2 mg/kg) did not lead to significant changes in body weight compared to the Vehicle Control group during the observation period (Fig. 12A), again suggesting that amlodipine was not toxic.

Unlike the positive control carrageenan, which showed significant paw edema in 24 hours, amlodipine had no effect and was comparable to control, as on day 1 (Fig.12 B). Over the course of treatment of 28 days, amlodipine showed no contribution towards either erythema or edema, and was comparable to the control (Fig 12C).

In some aspects, the foregoing description of the specific aspect will so fully reveal the general nature of the aspects herein that others can, by applying current knowledge, readily modify and/or adapt for various applications such specific aspect 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 at least one calcium channel blocker (CCBs) 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 2-16 mg, to a subject in need thereof.

2. The method according to claim 1, wherein the calcium channel blocker is selected from the group comprising non-dihydropyridines and dihydropyridines.

3. The method according to claim 2, wherein at least one calcium channel blocker is a dihydropyridine calcium antagonist.

4. The method according to claim 3, wherein at least one dihydropyridine calcium antagonist is Amlodipine.

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

6. The method according to claim 1, wherein the pharmaceutical salts are selected from the group comprising besylate, mesylate, maleate, and camsylate.

7. The method according to claim 1, 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.

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

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

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

11. The method according to claim 1, wherein 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.

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

13. The method according to claim 1, wherein the concentration of at least one calcium channel blocker in the composition may range from as low as 0.1% of the total amount of the composition up to as high as 100%.

14. The method according to claim 1, wherein the dosage of the composition is in the range of 2-16 mg per person per day.

15. The method according to claim 1, wherein the composition administered comprises 2-12 mg of amlodipine, pharmaceutically acceptable salts and therapeutic compounds.

16. The method according to claim 1, wherein the composition administered preferably comprises 10 mg of amlodipine.

17. A composition of delaying tumor metastasis comprises Amlodipine and pharmaceutically acceptable salt and/or in combination with one or more therapeutic compounds.

18. The composition according to claim 17, wherein the composition further comprises a pharmaceutically acceptable carrier, excipient, diluent and/or adjuvant.

19. The composition according to claim 17, wherein the therapeutic compound is selected from the group comprising losartan, propranolol, metformin, aspirin, atenolol, niclosomide and combinations thereof.

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

21. The composition according to claim 17, wherein the composition comprises Amlodipine and therapeutic compound in a ratio of 1:99.

22. The composition according to claim 17, 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.

23. The composition according to claim 17, wherein the composition comprises 2-12 mg of amlodipine, pharmaceutically acceptable salts and therapeutic compounds.

24. The composition according to claim 17, wherein the composition preferably comprises 10 mg of amlodipine.

25. Use of Amlodipine for delaying tumor metastasis comprising: a) preparing a composition comprising at least one calcium channel blocker (CCBs) and or pharmaceutically acceptable salt or in combination with one or more therapeutic compound; and b) the therapeutic effective amount in the range of 2- 16 mg is provided to a subject in need thereof.

26. The use according to claim 25, wherein the composition for delaying tumor metastasis comprises 2-12 mg of amlodipine, pharmaceutically acceptable salts and therapeutic compounds.

27. The use according to claim 25, wherein the composition for delaying tumor metastasis preferably comprises 10 mg of amlodipine.