FORMULATION AND METHOD FOR TARGETING MULTIDRUG-RESISTANT ENTEROBACTERIACEAE USING LIPOSOME-ENCAPSULATED TETRACYCLINE

Abstract

Disclosed herein is a formulation and method for targeting multidrug-resistant Enterobacteriaceae using liposome-encapsulated tetracycline (100) that comprises tetracycline as the primary active antibiotic agent, encapsulated to enable sustained, controlled release upon encountering the targeted bacteria, enhancing therapeutic efficacy while reducing systemic toxicity, phospholipids forming a bilayer around the encapsulated tetracycline, wherein the phospholipids are configured to create a stable and biocompatible membrane, which aids in controlled delivery and increases the formulation’s residence time in the bloodstream, cholesterol incorporated within the phospholipid bilayer, serving to increase membrane stability and rigidity, thereby prolonging the formulation’s integrity and functionality in physiological environments, surface-modifying ligands selected from a group of specific targeting molecules that bind to Enterobacteriaceae receptors, wherein the ligands enhance selective adherence to bacterial surfaces, a biocompatible polymer coating on the liposome surface, configured to evade immune recognition and extend circulation time.

Specification

Description:FIELD OF DISCLOSURE
[0001] The present disclosure relates to pharmaceutical compositions, more specifically, relates to formulation and method for targeting multidrug-resistant Enterobacteriaceae using liposome-encapsulated tetracycline.
BACKGROUND OF THE DISCLOSURE
[0002] The formulation is designed to deliver antibiotics directly to drug-resistant bacteria, helping to overcome efflux pump mediated drug resistance mechanisms. This targeted delivery can make treatments more efficient potentially reducing dosage needs and side effects compared to conventional antibiotics.
[0003] By encapsulating the antibiotic, the formulation helps stabilize the drug, preventing degradation in the body before it reaches the infection site. This can increase the duration of effectiveness, meaning that patients may need less frequent dosing.
[0004] Given its targeting of multidrug-resistant bacteria, the formulation has the potential to address a range of resistant bacterial strains. This can improve patient outcomes, especially for those who do not respond to standard treatments.
[0005] Some existing formulations may not deliver the antibiotic as precisely, resulting in less effective targeting of specific bacteria and potentially leading to off-target effects or damage to beneficial bacteria in the body.
[0006] Conventional antibiotic formulations without encapsulation often experience degradation in the bloodstream, leading to reduced efficacy by the time they reach the infection site. This can necessitate higher dosages, increasing side effects.
[0007] Non-encapsulated or non-targeted antibiotic therapies often circulate throughout the body, affecting both harmful and beneficial bacteria. This lack of specificity can lead to more pronounced side effects, such as gastrointestinal issues, and may reduce overall treatment adherence.
[0008] Thus, in light of the above-stated discussion, there exists a need for a formulation and method for targeting multidrug-resistant Enterobacteriaceae using liposome-encapsulated tetracycline.
SUMMARY OF THE DISCLOSURE
[0009] The following is a summary description of illustrative embodiments of the invention. It is provided as a preface to assist those skilled in the art to more rapidly assimilate the detailed design discussion which ensues and is not intended in any way to limit the scope of the claims which are appended hereto in order to particularly point out the invention.
[0010] According to illustrative embodiments, the present disclosure focuses on a formulation and method for targeting multidrug-resistant Enterobacteriaceae using liposome-encapsulated tetracycline which overcomes the above-mentioned disadvantages or provide the users with a useful or commercial choice.
[0011] An objective of the present disclosure is to formulate a stable antibiotic composition to enhance the efficacy of treatment against multidrug-resistant bacteria.
[0012] Another objective of the present disclosure is to provide a targeted formulation for delivering antibiotics directly to resistant bacterial infections, minimizing off-target effects.
[0013] Another objective of the present disclosure is to ensure the encapsulation of antibiotics within a biocompatible carrier that allows controlled release and extended drug stability within the body.
[0014] Another objective of the present disclosure is to reduce the frequency of antibiotic doses needed for effective treatment, supporting improved patient adherence and convenience.
[0015] Another objective of the present disclosure is to enhance the bioavailability of antibiotics by preventing premature degradation and ensuring delivery to the infection site.
[0016] Another objective of the present disclosure is to create a formulation which is capable of reducing formulationic side effects by enabling localized drug action.
[0017] Another objective of the present disclosure is to provide an antibiotic formulation that maintains efficacy across a variety of multidrug-resistant bacterial strains, broadening treatment options.
[0018] Another objective of the present disclosure is to develop a formulation with an optimized pharmacokinetic profile, promoting sustained antibiotic action at therapeutic levels.
[0019] Another objective of the present disclosure is to address bacterial resistance mechanisms through an advanced delivery formulation that enhances antibiotic interaction with target bacteria.
[0020] Yet another objective of the present disclosure is to improve the safety profile of antibiotic treatments, particularly for patients requiring prolonged or repeated doses, by minimizing exposure of non-target areas to the drug.
[0021] In light of the above, in one aspect of the present disclosure, a formulation for targeting multidrug-resistant Enterobacteriaceae using liposome-encapsulated tetracycline is disclosed herein. The formulation comprises tetracycline as the primary active antibiotic agent, encapsulated to enable sustained, controlled release upon encountering the targeted bacteria, enhancing therapeutic efficacy while reducing systemic toxicity. The formulation includes phospholipids forming a bilayer around the encapsulated tetracycline, wherein the phospholipids are configured to create a stable and biocompatible membrane, which aids in controlled delivery and increases the formulation's residence time in the bloodstream. The formulation also includes cholesterol incorporated within the phospholipid bilayer, serving to increase membrane stability and rigidity, thereby prolonging the formulation's integrity and functionality in physiological environments. and extend circulation time, thus maximizing contact with the target bacterial cells within an infected host.
[0022] In one embodiment, the tetracycline is present in an amount of 5-10% by weight of the formulation, optimizing the antibiotic's encapsulation efficiency and sustained release.
[0023] In one embodiment, the phospholipids are present in an amount of 50-70% by weight, forming a stable bilayer that allows for effective encapsulation and controlled release of tetracycline.
[0024] In one embodiment, the cholesterol is present in an amount of 20-30% by weight, contributing to the liposome's membrane rigidity and enhancing stability in physiological conditions.
[0025] In one embodiment, the surface-modifying ligands are present at 1-5% by weight, configured to enhance targeting specificity towards Enterobacteriaceae by binding to bacterial surface receptors.
[0026] In one embodiment, the liposomes have a particle size ranging from 100-200 nanometres, facilitating enhanced cellular uptake by Enterobacteriaceae and penetration into biofilms.
[0027] In one embodiment, the surface of the liposomes is positively charged, achieving an optimal zeta potential to enhance interactions with the negatively charged bacterial cell membranes.
[0028] In one embodiment, the encapsulation efficiency of tetracycline is maintained between 70-90%, ensuring prolonged release and increased therapeutic efficacy against multidrug-resistant Enterobacteriaceae.
[0029] In light of the above, in one aspect of the present disclosure, a method for targeting multidrug-resistant Enterobacteriaceae using liposome-encapsulated tetracycline is disclosed herein. The method comprises encapsulating tetracycline within a liposomal structure, wherein the tetracycline is introduced as the primary antibiotic agent for controlled release upon interaction with the target bacteria, thereby enhancing therapeutic efficacy while minimizing systemic toxicity. The method includes forming a bilayer around the encapsulated tetracycline using phospholipids, configuring the phospholipids to create a stable, biocompatible membrane that allows sustained release and prolongs residence time in the bloodstream. The method also includes incorporating cholesterol within the phospholipid bilayer, wherein the cholesterol improves membrane rigidity and stability, enhancing the overall integrity and performance of the liposome in physiological conditions.
[0030] These and other advantages will be apparent from the present application of the embodiments described herein.
[0031] The preceding is a simplified summary to provide an understanding of some embodiments of the present invention. This summary is neither an extensive nor exhaustive overview of the present invention and its various embodiments. The summary presents selected concepts of the embodiments of the present invention in a simplified form as an introduction to the more detailed description presented below. As will be appreciated, other embodiments of the present invention are possible utilizing, alone or in combination, one or more of the features set forth above or described in detail below.
[0032] These elements, together with the other aspects of the present disclosure and various features are pointed out with particularity in the claims annexed hereto and form a part of the present disclosure. For a better understanding of the present disclosure, its operating advantages, and the specified object attained by its uses, reference should be made to the accompanying drawings and descriptive matter in which there are illustrated exemplary embodiments of the present disclosure.
BRIEF DESCRIPTION OF THE DRAWINGS
[0033] To describe the technical solutions in the embodiments of the present disclosure or in the prior art more clearly, the following briefly describes the accompanying drawings required for describing the embodiments or the prior art. Apparently, the accompanying drawings in the following description merely show some embodiments of the present disclosure, and a person of ordinary skill in the art can derive other implementations from these accompanying drawings without creative efforts. All of the embodiments or the implementations shall fall within the protection scope of the present disclosure.
[0034] The advantages and features of the present disclosure will become better understood with reference to the following detailed description taken in conjunction with the accompanying drawing, in which:
[0035] FIG. 1 illustrates a flowchart of a formulation for targeting multidrug-resistant Enterobacteriaceae using liposome-encapsulated tetracycline, in accordance with an exemplary embodiment of the present disclosure;
[0036] FIG. 2 illustrates a flowchart of a method for targeting multidrug-resistant Enterobacteriaceae using liposome-encapsulated tetracycline, in accordance with an exemplary embodiment of the present disclosure;
[0037] FIG. 3 illustrates a screenshot of a representative graphical image showing particle size, PI and zeta potential of liposomes with tetracycline encapsulation, in accordance with an exemplary embodiment of the present disclosure;
[0038] FIG. 4 illustrates a screenshot of a TEM image showing the interaction of tetracycline encapsulated liposome on the bacterial surface, in accordance with an exemplary embodiment of the present disclosure;
[0039] FIG. 5A to 5F illustrates a perspective view of the MHA plates showing zone of inhibition in presence of tetracycline formulation in KP33, KP03 and KP22 as control plate with TE disc, 30μg and 10mg of free TE, and plain liposome and also control plate with different volume of TE encapsulated liposome, in accordance with an exemplary embodiment of the present disclosure;
[0040] FIG. 6A to 6C illustrates a perspective view of the MHA plates showing zone of inhibition in KP33, KP03 and KP22 plates showing 10 μg free antibiotic as control and different TE-liposome formulation;
[0041] FIG. 7 illustrates a screenshot of a MIC determination, isolates showing clearance at 10.3 μg/ml as MIC for the TE-Liposome formulation;
[0042] FIG. 8A to 8B illustrates a perspective view of the step-by-step of the working model of the formulation for targeting multidrug-resistant Enterobacteriaceae using liposome-encapsulated tetracycline.
[0043] Like reference, numerals refer to like parts throughout the description of several views of the drawing.
[0044] The formulation and method for targeting multidrug-resistant Enterobacteriaceae using liposome-encapsulated tetracycline is illustrated in the accompanying drawings, which like reference letters indicate corresponding parts in the various figures. It should be noted that the accompanying figure is intended to present illustrations of exemplary embodiments of the present disclosure. This figure is not intended to limit the scope of the present disclosure. It should also be noted that the accompanying figure is not necessarily drawn to scale.
DETAILED DESCRIPTION OF THE DISCLOSURE
[0045] The following is a detailed description of embodiments of the disclosure depicted in the accompanying drawings. The embodiments are in such detail as to communicate the disclosure. However, the amount of detail offered is not intended to limit the anticipated variations of embodiments; on the contrary, the intention is to cover all modifications, equivalents, and alternatives falling within the spirit and scope of the present disclosure.
[0046] In the following description, numerous specific details are set forth in order to provide a thorough understanding of the embodiments of the present disclosure. It may be apparent to one skilled in the art that embodiments of the present disclosure may be practiced without some of these specific details.
[0047] Various terms as used herein are shown below. To the extent a term is used, it should be given the broadest definition persons in the pertinent art have given that term as reflected in printed publications and issued patents at the time of filing.
[0048] The terms "a" and "an" herein do not denote a limitation of quantity, but rather denote the presence of at least one of the referenced items.
[0049] The terms "having", "comprising", "including", and variations thereof signify the presence of a component.
[0050] Referring now to FIG. 1 to FIG. 8 to describe various exemplary embodiments of the present disclosure. FIG. 1 illustrates flowchart of a formulation for targeting multidrug-resistant Enterobacteriaceae using liposome-encapsulated tetracycline 100, in accordance with an exemplary embodiment of the present disclosure.
[0051] The formulation 100 may include tetracycline as the primary active antibiotic agent, encapsulated to enable sustained, controlled release upon encountering the targeted bacteria, enhancing therapeutic efficacy while reducing systemic toxicity. The formulation 100 may also include phospholipids forming a bilayer around the encapsulated tetracycline, wherein the phospholipids are configured to create a stable and biocompatible membrane, which aids in controlled delivery and increases the formulation's residence time in the bloodstream. The formulation 100 may also include cholesterol incorporated within the phospholipid bilayer, serving to increase membrane stability and rigidity, thereby prolonging the formulation's integrity and functionality in physiological environments.
[0052] The tetracycline is present in an amount of 5-10% by weight of the formulation, optimizing the antibiotic's encapsulation efficiency and sustained release.
[0053] The phospholipids are present in an amount of 50-70% by weight, forming a stable bilayer that allows for effective encapsulation and controlled release of tetracycline.
[0054] The cholesterol is present in an amount of 20-30% by weight, contributing to the liposome's membrane rigidity and enhancing stability in physiological conditions.
[0055] The surface-modifying ligands are present at 1-5% by weight, configured to enhance targeting specificity towards Enterobacteriaceae by binding to bacterial surface receptors.
[0056] The liposomes have a particle size ranging from 100-200 nanometres, facilitating enhanced cellular uptake by Enterobacteriaceae and penetration into biofilms.
[0057] The surface of the liposomes is positively charged, achieving an optimal zeta potential to enhance interactions with the negatively charged bacterial cell membranes.
[0058] The encapsulation efficiency of tetracycline is maintained between 70-90%, ensuring prolonged release and increased therapeutic efficacy against multidrug-resistant Enterobacteriaceae.
[0059] The method 100 may include encapsulating tetracycline within a liposomal structure, wherein the tetracycline is introduced as the primary antibiotic agent for controlled release upon interaction with the target bacteria, thereby enhancing therapeutic efficacy while minimizing systemic toxicity. The method 100 may also include forming a bilayer around the encapsulated tetracycline using phospholipids, configuring the phospholipids to create a stable, biocompatible membrane that allows sustained release and prolongs residence time in the bloodstream. The method 100 may also include incorporating cholesterol within the phospholipid bilayer, wherein the cholesterol improves membrane rigidity and stability, enhancing the overall integrity and performance of the liposome in physiological conditions.
[0060] At 102, prepare a transparent, flexible polymer substrate as the foundation for the formulation.
[0061] At 104, layer a controlled amount of tetracycline as the primary active agent, encapsulating it for sustained release.
[0062] At 106, assemble phospholipids around the tetracycline to create a stable, biocompatible bilayer membrane.
[0063] At 108, integrate cholesterol within the phospholipid bilayer to enhance structural integrity in physiological environments.
[0064] FIG. 2 illustrates a flowchart of a method for targeting multidrug-resistant Enterobacteriaceae using liposome-encapsulated tetracycline, in accordance with an exemplary embodiment of the present disclosure.
[0065] At 202, encapsulating tetracycline within a liposomal structure, wherein the tetracycline is introduced as the primary antibiotic agent for controlled release upon interaction with the target bacteria, thereby enhancing therapeutic efficacy while minimizing systemic toxicity.
[0066] At 204, forming a bilayer around the encapsulated tetracycline using phospholipids, configuring the phospholipids to create a stable, biocompatible membrane that allows sustained release and prolongs residence time in the bloodstream.
[0067] At 206, incorporating cholesterol within the phospholipid bilayer, wherein the cholesterol improves membrane rigidity and stability, enhancing the overall integrity and performance of the liposome in physiological conditions.
[0068] FIG. 3 illustrates a screenshot of a representative graphical image showing particle size, PI and zeta potential of liposomes with tetracycline encapsulation, in accordance with an exemplary embodiment of the present disclosure.
[0069] The graphical image illustrates key properties of the liposomes with tetracycline encapsulation, showing particle size, polydispersity index (PI), and zeta potential. The particle size data represents the formulation's optimized dimensions for efficient cellular uptake, while the PI indicates uniformity in particle distribution. The zeta potential measurement reflects the surface charge, crucial for stability and interaction with bacterial cells in therapeutic applications.
[0070] FIG. 4 illustrates a screenshot of a TEM image showing the [1] tetracycline encapsulated liposome, [2] interaction of tetracycline encapsulated liposome with the bacterial, [3] Bacterial cells have liposomes surrounding them in accordance with an exemplary embodiment of the present disclosure.
[0071] The TEM image shows direct interaction between tetracycline-encapsulated liposomes and the bacterial surface, illustrating effective adhesion essential for targeted antibiotic delivery. The liposome attachment to the bacteria demonstrates the formulation's design for selective binding and delivery, enhancing localized tetracycline release. This interaction underpins the formulation's goal of maximizing efficacy against multidrug-resistant Enterobacteriaceae.
[0072] FIG. 5A to 5F illustrates a perspective view of the MHA plates showing zone of inhibition in presence of tetracycline formulation in KP33, KP03 and KP22 as control plate with TE disc, 30μg and 10mg of free TE, and plain liposome and also control plate with different volume of TE encapsulated liposome, in accordance with an exemplary embodiment of the present disclosure.
[0073] FIG. 5A illustrates a perspective view of the MHA plates showing zone of inhibition in presence of tetracycline formulation in KP33 as control plate with TE disc, 30μg and 10mg of free TE, and plain liposome, in accordance with an exemplary embodiment of the present disclosure.
[0074] FIG. 5B illustrates a perspective view of the MHA plates showing zone of inhibition in presence of tetracycline formulation in KP33 as control plate with different volume of TE encapsulated liposome, in accordance with an exemplary embodiment of the present disclosure.
[0075] FIG. 5C illustrates a perspective view of the MHA plates showing zone of inhibition in presence of tetracycline formulation in KP03 as control plate with TE disc, 30μg and 10mg of free TE, and plain liposome, in accordance with an exemplary embodiment of the present disclosure.
[0076] FIG. 5D illustrates a perspective view of the MHA plates showing zone of inhibition in presence of tetracycline formulation in KP03 as control plate with different volume of TE encapsulated liposome, in accordance with an exemplary embodiment of the present disclosure.
[0077] FIG. 5E illustrates a perspective view of the MHA plates showing zone of inhibition in presence of tetracycline formulation in KP22 as control plate with TE disc, 30μg and 10mg of free TE, and plain liposome, in accordance with an exemplary embodiment of the present disclosure.
[0078] FIG. 5F illustrates a perspective view of the MHA plates showing zone of inhibition in presence of tetracycline formulation in KP22 as control plate with different volume of TE encapsulated liposome, in accordance with an exemplary embodiment of the present disclosure.
[0079] FIG. 6A to 6C illustrates a perspective view of the MHA plates showing zone of inhibition in KP33, KP03 and KP22 plates showing 10 μg free antibiotic as control and different TE-liposome formulation, in accordance with an exemplary embodiment of the present disclosure.
[0080] FIG. 6A illustrates a perspective view of the MHA plates showing zone of inhibition in KP33 plates showing 10 μg free antibiotic as control and different TE-liposome formulation, in accordance with an exemplary embodiment of the present disclosure.
[0081] FIG. 6B illustrates a perspective view of the MHA plates showing zone of inhibition in KP03 plates showing 10 μg free antibiotic as control and different TE-liposome formulation, in accordance with an exemplary embodiment of the present disclosure.
[0082] FIG. 6C illustrates a perspective view of the MHA plates showing zone of inhibition in KP22 plates showing 10 μg free antibiotic as control and different TE-liposome formulation, in accordance with an exemplary embodiment of the present disclosure.
[0083] FIG. 7 illustrates a screenshot of a MIC determination, isolates showing clearance at 10.3 μg/ml as MIC for the TE-Liposome formulation, in accordance with an exemplary embodiment of the present disclosure.
[0084] The MIC determination image illustrates the effectiveness of the TE-Liposome formulation, with bacterial clearance achieved at a concentration of 10.3 μg/ml. This indicates the minimum inhibitory concentration required for the TE-Liposome formulation to inhibit bacterial growth, demonstrating its potency against the targeted multidrug-resistant Enterobacteriaceae. The image highlights the formulation's ability to reach therapeutic thresholds with low antibiotic concentrations, supporting targeted and efficient antibacterial action.
[0085] FIG. 8A to 8B illustrates a perspective view of the step-by-step of the working model of the formulation for targeting multidrug-resistant Enterobacteriaceae using liposome-encapsulated tetracycline, in accordance with an exemplary embodiment of the present disclosure.
[0086] Rotary flash evaporator 802 is a specialized device used for the continuous and controlled removal of solvents from lipid solutions. During the operation of rotary flash evaporator 802, a vacuum environment reduces the boiling point of solvents, allowing them to evaporate at lower temperatures, thus preserving the integrity of sensitive lipid compounds. Rotary flash evaporator 802 operates by rotating the lipid solution within a heated vessel, creating a thin film along the inner surface, which enables rapid and efficient evaporation of solvents. This process results in a concentrated lipid residue that is evenly distributed on the container surface, known as the dry lipid layer 804. The creation of dry lipid layer 804 is essential for the subsequent steps, as it provides the basis for forming stable liposomes. The consistency and homogeneity achieved by rotary flash evaporator 802 contribute to the quality of the final liposome product by ensuring a well-structured lipid base. Additionally, rotary flash evaporator 802 enhances the purity of lipids by continuously removing any remaining traces of volatile solvents, which could otherwise compromise the stability of liposomes during encapsulation. The uniformity of the lipid film produced by rotary flash evaporator 802 directly influences the efficiency of encapsulation and the overall quality of the liposomal formulation, making rotary flash evaporator 802 an indispensable component in the process of tetracycline encapsulation.
[0087] Dry lipid layer 804 is a thin and concentrated film of lipids that forms on the inner surface of the container following solvent removal by rotary flash evaporator 802. The creation of dry lipid layer 804 is a crucial step for successful liposome formation, as it provides a structured matrix for hydration and subsequent encapsulation of tetracycline. Dry lipid layer 804 serves as the foundational structure upon which liposomes are built, ensuring that the lipid molecules are arranged in a manner conducive to forming stable bilayered vesicles. The thickness and uniformity of dry lipid layer 804 are carefully controlled to ensure consistency in liposome size and encapsulation efficiency. In addition to providing a matrix for hydration, dry lipid layer 804 also helps to stabilize the lipids by preventing premature aggregation, which can lead to variability in the final liposomal product. When aqueous solutions are introduced in hydration 806, dry lipid layer 804 swells and transforms into bilayer structures that encapsulate the tetracycline molecules, enhancing their stability and targeting capabilities. Thus, the quality and consistency of dry lipid layer 804 directly impact the effectiveness of the liposomal formulation in delivering tetracycline to bacterial cells.
[0088] Hydration 806 is the process of introducing an aqueous solution to dry lipid layer 804, causing it to swell and form bilayered vesicular structures, which are the precursors to liposomes 808. The addition of an aqueous medium during hydration 806 enables lipid molecules within dry lipid layer 804 to self-assemble into spherical bilayers, forming enclosed spaces that can effectively encapsulate tetracycline. This process is meticulously controlled to ensure that the resulting liposomes 808 are uniform in size, which is critical for consistent drug delivery. The effectiveness of hydration 806 relies on precise temperature and pH conditions, which facilitate the optimal swelling and transformation of dry lipid layer 804 into a stable, encapsulated structure. By maintaining these conditions, hydration 806 ensures that liposomes 808 retain their structural integrity and are capable of carrying tetracycline to bacterial cells without premature release. The hydration process also contributes to the long-term stability of the liposomes, allowing for a controlled and sustained release of tetracycline at the target site.
[0089] Liposomes 808 are the encapsulated vesicular structures formed through hydration 806 of dry lipid layer 804. These bilayered vesicles play a central role in the targeted delivery of tetracycline, as they provide a protective environment that enhances both stability and bioavailability of the drug. Liposomes 808 consist of lipid bilayers that encapsulate tetracycline molecules, shielding them from external environmental factors that could otherwise degrade the drug. The structural integrity of liposomes 808 is essential for ensuring controlled release, allowing for a gradual and sustained release of tetracycline at the bacterial target site. The design of liposomes 808 enables them to interact effectively with bacterial cells, promoting efficient drug uptake and enhancing antibacterial efficacy. By encapsulating tetracycline within liposomes 808, the formulation minimizes adverse effects and maximizes the therapeutic impact by directing the drug specifically to the bacterial site.
[0090] Thin film hydration technique 810 is a method that integrates rotary flash evaporator 802 with hydration 806 to achieve the formation of liposomes 808 with controlled characteristics. Thin film hydration technique 810 involves a multi-step process wherein lipids are first dissolved and distributed evenly by rotary flash evaporator 802, forming dry lipid layer 804. Following this, hydration 806 is applied to transform dry lipid layer 804 into liposomes 808 with desired size and encapsulation properties. Thin film hydration technique 810 is widely recognized for its ability to produce liposomes with high encapsulation efficiency, making it a preferred method in pharmaceutical applications involving drug delivery systems. By controlling the parameters of thin film hydration technique 810, researchers can produce liposomes with consistent size distribution, stability, and encapsulation capacity, which are essential for effective tetracycline delivery.
[0091] Sonicator 812 is an ultrasonic device that applies sound waves to liposomes 808, reducing their particle size and achieving uniform dispersion within the suspension. The application of ultrasonic energy by sonicator 812 creates microbubbles in the solution, which collapse and generate shear forces that break down larger liposomal aggregates. This process is crucial for producing liposomes 808 of a uniform size, which is important for ensuring consistent drug delivery and bioavailability. The operation of sonicator 812 also prevents liposome aggregation, which could compromise the stability and effectiveness of the liposomal formulation. By ensuring a homogeneous size distribution, sonicator 812 enhances the therapeutic potential of tetracycline encapsulated within liposomes 808, enabling better interaction with bacterial cells and improved absorption.
[0092] Zeta analyser 814 is an analytical instrument that measures the zeta potential and particle size of liposomes 808, providing essential data on their stability and dispersion properties. The measurement of zeta potential by zeta analyser 814 helps determine the surface charge of liposomes 808, which influences their interaction with bacterial cells and resistance to aggregation. A stable zeta potential is indicative of a well-dispersed liposome suspension, which is critical for maintaining the integrity and effectiveness of the encapsulated drug. By analysing these properties, zeta analyser 814 allows for fine-tuning of the formulation to ensure that liposomes 808 are optimally prepared for targeted drug delivery applications.
[0093] Transmission electron microscopy (TEM) analysis 816 is a high-resolution imaging technique used to observe the structural characteristics of tetracycline-encapsulated liposomes 834. TEM analysis 816 provides visual confirmation of the size, morphology, and encapsulation quality of the liposomes, ensuring they meet the necessary standards for effective drug delivery. By examining the ultrastructure of tetracycline-encapsulated liposomes 834, TEM analysis 816 helps verify that the liposomes are well-suited for interaction with bacterial cells, supporting their stability and efficacy in therapeutic applications. This analysis also allows for the detection of any structural anomalies, ensuring that the liposomes are consistent in their size and encapsulation properties.
[0094] Agar well diffusion assay 818 is a laboratory technique used to evaluate the antibacterial activity of tetracycline-encapsulated liposomes 834. During agar well diffusion assay 818, the liposomal formulation is introduced into wells on an agar plate inoculated with bacterial cultures. The effectiveness of the formulation is measured by observing the zones of inhibition around each well, indicating the extent of bacterial suppression. Agar well diffusion assay 818 is a direct measure of the formulation's potential to inhibit bacterial growth, providing valuable data on its therapeutic efficacy.
[0095] Adhesion process 820 refers to the binding interaction between tetracycline-encapsulated liposomes 834 and the bacterial cell membrane. The adhesion process 820 is critical as it facilitates the targeted delivery of tetracycline by ensuring effective attachment of liposomes to bacterial cells. This interaction allows for precise localization of tetracycline, enhancing its antibacterial impact while minimizing off-target effects. The adhesion process 820 supports the controlled release of tetracycline upon contact with bacterial cells, ensuring the therapeutic agent is delivered exactly where it is needed.
[0096] The fusion of liposome 822 with bacterial cell membrane 824 is a carefully engineered process designed to deliver tetracycline encapsulated lipoids 834 directly to bacterial cells. Liposome 822 aligns with bacterial cell membrane 824, facilitating direct entry and enabling efficient therapeutic action within bacterial cytoplasm 836. This interaction ensures that tetracycline encapsulated lipoids 834 bypass natural bacterial defences, allowing the therapeutic compound to exert its full potential upon release. The lipids in liposome 822 are optimized for compatibility with bacterial cell membrane 824, maintaining stability and enhancing the efficacy of the encapsulated drug. This fusion process ensures controlled and targeted delivery of tetracycline encapsulated lipoids 834, minimizing the risk of drug degradation and supporting sustained interaction with bacterial ribosomes. As a result, the controlled release mechanism of tetracycline within bacterial cytoplasm 836 enhances its therapeutic effects, ultimately contributing to the successful inhibition of bacterial growth and replication.
[0097] The selective permeability of bacterial cell membrane 824 is an essential characteristic that bacterial cells use to regulate the entry and exit of molecules. Bacterial cell membrane 824 distinguishes between beneficial nutrients and potentially harmful substances, which is critical for bacterial survival. Liposome 822, however, is specifically designed to align with the lipid structure of bacterial cell membrane 824, allowing it to bypass this selective barrier and achieve effective fusion. Through this design, liposome 822 can deliver tetracycline encapsulated lipoids 834 into bacterial cytoplasm 836, enhancing drug accumulation in the target area. This mechanism helps to ensure that tetracycline reaches bacterial ribosomes without interference from efflux mechanisms. By bypassing natural cellular barriers, liposome 822 achieves targeted and efficient drug delivery, reducing off-target effects and enhancing the effectiveness of tetracycline encapsulated lipoids 834 within bacterial cytoplasm 836.
[0098] The bypass of efflux mechanism 826 in bacterial cells represents a critical function that enables tetracycline encapsulated lipoids 834 to reach bacterial cytoplasm 836 effectively. Efflux mechanisms are defence systems in bacterial cell membrane 824 designed to expel foreign substances, such as antibiotics, from bacterial cells, thereby reducing the drug's intracellular concentration and limiting its therapeutic impact. Tetracycline encapsulated lipoids 834 are formulated to avoid these bacterial defences, leveraging the structural design of liposome 822 to facilitate entry into bacterial cells without activating the efflux pump.
[0099] Bypassing efflux mechanism 826 ensures that tetracycline encapsulated lipoids 834 can deliver their therapeutic load directly to bacterial cytoplasm 836. This process relies on the compatibility of liposome 822 with bacterial cell membrane 824, which allows for seamless fusion and circumvention of natural expulsion processes. The careful engineering of liposome 822 is instrumental in minimizing the likelihood of efflux, thereby ensuring that tetracycline encapsulated lipoids 834 retain a high concentration of tetracycline within bacterial cytoplasm 836. By avoiding premature expulsion, tetracycline encapsulated lipoids 834 achieve optimal therapeutic efficacy, targeting bacterial ribosomes with sustained potency.
[0100] The binding of tetracycline 828 to bacterial ribosomes within bacterial cytoplasm 836 is a critical process that underpins the effectiveness of the antibiotic. Tetracycline 828 exhibits a strong affinity for the smaller ribosomal subunits present in bacterial cells, effectively blocking sites that are essential for protein synthesis. By engaging directly with bacterial ribosomes, tetracycline 828 prevents the translation of bacterial mRNA into functional proteins, thereby hindering bacterial replication and growth. This precise interaction enables tetracycline encapsulated lipoids 834 to maintain selective antibacterial activity, ensuring minimal impact on surrounding non-target cells. The specificity of binding of tetracycline 828 allows the drug to achieve a high level of therapeutic precision, effectively managing bacterial infections by targeting ribosomal components unique to bacteria. This specific binding interaction represents a significant advantage in enhancing the efficacy of tetracycline encapsulated lipoids 834.
[0101] The inhibition of protein synthesis 830 in bacterial cytoplasm 836 is the outcome of successful binding of tetracycline 828 to bacterial ribosomes. By blocking protein synthesis, tetracycline 828 effectively stops bacterial cells from generating essential proteins, which are crucial for survival and replication. This inhibition of protein synthesis 830 directly impacts bacterial growth, as bacterial cells are unable to perform essential functions. Tetracycline encapsulated lipoids 834 deliver a sustained concentration of the antibiotic within bacterial cytoplasm 836, ensuring prolonged inhibition of protein synthesis. This precise inhibition mechanism is facilitated by the targeted release of tetracycline encapsulated lipoids 834, which enhances the drug's therapeutic efficacy while minimizing collateral effects on host cells. By focusing on bacterial cells, tetracycline encapsulated lipoids 834 ensure a targeted approach to managing bacterial infections, effectively contributing to cell death.
[0102] The release of tetracycline 832 into bacterial cytoplasm 836 occurs after successful fusion of liposome 822 with bacterial cell membrane 824. This release mechanism ensures that tetracycline encapsulated lipoids 834 remain intact until they reach bacterial cytoplasm 836. By maintaining stability of tetracycline within liposome 822 during transport, this release process facilitates controlled dispersion of tetracycline within bacterial cytoplasm 836. The release of tetracycline 832 enables direct interaction with bacterial ribosomes, supporting prolonged inhibition of bacterial protein synthesis. This controlled release mechanism maximizes the drug's effectiveness within bacterial cells by ensuring that tetracycline encapsulated lipoids 834 have a sustained presence. By enabling precise delivery and controlled release, tetracycline encapsulated lipoids 834 are optimized for enhanced antibacterial action against target bacterial cells.
[0103] Tetracycline encapsulated lipoids 834 serve as essential carriers for delivering tetracycline directly to bacterial cells, protecting the drug from premature degradation and ensuring it bypasses bacterial defence mechanisms. By encapsulating tetracycline within liposome 822, tetracycline encapsulated lipoids 834 allow the drug to avoid interaction with efflux mechanisms commonly found in bacterial cell membrane 824. Upon fusion with bacterial cell membrane 824, tetracycline encapsulated lipoids 834 deliver tetracycline into bacterial cytoplasm 836, where it can effectively interact with bacterial ribosomes. This encapsulation design significantly enhances the bioavailability of tetracycline within bacterial cells, ensuring the drug retains its potency for effective therapeutic action. By providing a stable and protective environment, tetracycline encapsulated lipoids 834 enhance the efficacy of tetracycline in targeting bacterial infections.
[0104] The bacterial cytoplasm 836 serves as the primary site where tetracycline encapsulated lipoids 834 interact with bacterial ribosomes to inhibit protein synthesis. Upon reaching bacterial cytoplasm 836, tetracycline is released from liposome 822 in a controlled manner, allowing sustained interaction with bacterial ribosomes. This targeted delivery mechanism enhances drug accumulation in bacterial cytoplasm 836, ensuring effective binding of tetracycline 828 to ribosomal components. Bacterial cytoplasm 836 provides an environment that allows tetracycline to inhibit essential protein synthesis processes, ultimately leading to cell death. By achieving targeted delivery to bacterial cytoplasm 836, tetracycline encapsulated lipoids 834 maximize therapeutic effects while minimizing off-target interactions. The strategic design of liposome 822 ensures that bacterial cytoplasm 836 becomes the focal point for tetracycline's antibacterial action.
[0105] While the invention has been described in connection with what is presently considered to be the most practical and various embodiments, it will be understood that the invention is not to be limited to the disclosed embodiments, but on the contrary, is intended to cover various modifications and equivalent arrangements included within the scope of the appended claims. A person of ordinary skill in the art may be aware that, in combination with the examples described in the embodiments disclosed in this specification, units and algorithm steps may be implemented by electronic hardware, computer software, or a combination thereof.
[0106] The foregoing descriptions of specific embodiments of the present disclosure have been presented for purposes of illustration and description. They are not intended to be exhaustive or to limit the present disclosure to the precise forms disclosed, and many modifications and variations are possible in light of the above teaching. The embodiments were chosen and described to best explain the principles of the present disclosure and its practical application, and to thereby enable others skilled in the art to best utilize the present disclosure and various embodiments with various modifications as are suited to the particular use contemplated. It is understood that various omissions and substitutions of equivalents are contemplated as circumstances may suggest or render expedient, but such omissions and substitutions are intended to cover the application or implementation without departing from the scope of the present disclosure.
[0107] Disjunctive language such as the phrase "at least one of X, Y, Z," unless specifically stated otherwise, is otherwise understood with the context as used in general to present that an item, term, etc., may be either X, Y, or Z, or any combination thereof (e.g., X, Y, and/or Z). Thus, such disjunctive language is not generally intended to, and should not, imply that certain embodiments require at least one of X, at least one of Y, or at least one of Z to each be present.
[0108] In a case that no conflict occurs, the embodiments in the present disclosure and the features in the embodiments may be mutually combined. The foregoing descriptions are merely specific implementations of the present disclosure, but are not intended to limit the protection scope of the present disclosure. Any variation or replacement readily figured out by a person skilled in the art within the technical scope disclosed in the present disclosure shall fall within the protection scope of the present disclosure. Therefore, the protection scope of the present disclosure shall be subject to the protection scope of the claims.
, Claims:I/We Claim:
1. A formulation for targeting multidrug-resistant Enterobacteriaceae using liposome-encapsulated tetracycline (100) comprising:
tetracycline as the primary active antibiotic agent, encapsulated to enable sustained, controlled release upon encountering the targeted bacteria, enhancing therapeutic efficacy while reducing systemic toxicity;
phospholipids forming a bilayer around the encapsulated tetracycline, wherein the phospholipids are configured to create a stable and biocompatible membrane, which aids in controlled delivery and increases the formulation's residence time in the bloodstream;
cholesterol incorporated within the phospholipid bilayer, serving to increase membrane stability and rigidity, thereby prolonging the formulation's integrity and functionality in physiological environments;
surface-modifying ligands selected from a group of specific targeting molecules that bind to Enterobacteriaceae receptors, wherein the ligands enhance selective adherence to bacterial surfaces, increasing uptake of the antibiotic by the targeted bacteria;
2. The formulation (100) as claimed in claim 1, wherein the tetracycline is present in an amount of 5-10% by weight of the formulation, optimizing the antibiotic's encapsulation efficiency and sustained release.
3. The formulation (100) as claimed in claim 1, wherein the phospholipids are present in an amount of 50-70% by weight, forming a stable bilayer that allows for effective encapsulation and controlled release of tetracycline.
4. The formulation (100) as claimed in claim 1, wherein the cholesterol is present in an amount of 20-30% by weight, contributing to the liposome's membrane rigidity and enhancing stability in physiological conditions.
5. The formulation (100) as claimed in claim 1, wherein the surface-modifying ligands are present at 1-5% by weight, configured to enhance targeting specificity towards Enterobacteriaceae by binding to bacterial surface receptors.
6. The formulation (100) as claimed in claim 1, wherein the liposomes have a particle size ranging from 100-200 nanometres, facilitating enhanced cellular uptake by Enterobacteriaceae and penetration into biofilms.
7. The formulation (100) claimed in claim 1, wherein the surface of the liposomes is positively charged, achieving an optimal zeta potential to enhance interactions with the negatively charged bacterial cell membranes.
8. The formulation (100) as claimed in claim 1, wherein the encapsulation efficiency of tetracycline is maintained between 70-90%, ensuring prolonged release and increased therapeutic efficacy against multidrug-resistant Enterobacteriaceae.
9. A method for targeting multidrug-resistant Enterobacteriaceae using liposome-encapsulated tetracycline (400) comprising:
encapsulating tetracycline within a liposomal structure, wherein the tetracycline is introduced as the primary antibiotic agent for controlled release upon interaction with the target bacteria, thereby enhancing therapeutic efficacy while minimizing systemic toxicity;
forming a bilayer around the encapsulated tetracycline using phospholipids, configuring the phospholipids to create a stable, biocompatible membrane that allows sustained release and prolongs residence time in the bloodstream;
incorporating cholesterol within the phospholipid bilayer, wherein the cholesterol improves membrane rigidity and stability, enhancing the overall integrity and performance of the liposome in physiological conditions.

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