An Upgraded Review: Anti-Diabetic Medication administration by Liposomal Formulation

Vikas D. Nikam*, Mitesh P. Sonawane, Kirti S. Pawar, Akash Rathod, Akash Tamboli

Department of Pharmaceutics, Loknete Dr. J. D. Pawar College of Pharmacy, Manur, Kalwan, Maharashtra.

*Corresponding Author E-mail: vikasnikam6616@gmail.com

ABSTRACT:

The prevalence of diabetes mellitus is increasing globally. Individuals suffering from diabetic complications encounter a range of social, mental, and physical challenges. Liposomes have been developed as a nanocarrier to enhance the administration of oral hypoglycaemic medications, surpassing the efficacy of conventional therapy. Liposomes offer significant regulation of raised blood glucose levels, rendering them a highly intriguing and potential therapeutic approach in contemporary research. Liposomes serve as a protective barrier for encapsulated hypoglycaemic medicines, hence optimising the efficacy of prolonged reductions in blood glucose levels. Hence, the objective of enhancing diabetes management while minimising the likelihood of both immediate and long-term complications is facilitated by the utilisation of liposomes filled with hypoglycaemic drugs. This article begins by examining the present condition of diabetes mellitus and the essential components of liposomal medicine delivery systems. We will next go into great detail about the composition, production methods, structure, along with drug incorporation of liposomes. In order to assure the efficiency of liposomes in both in vitro and in vivo settings, it is imperative to thoroughly characterise them prior to their utilisation. Consequently, numerous attributes of liposomes were examined. The primary objective of this review is to examine the limitations and stability of liposomes.

KEYWORDS: Antidiabetic, Liposome, Phospholipids, Drug Delivery, Liposome preparation, Liposome characterization.

1. INTRODUCTION:

Diabetes mellitus is a prevalent metabolic disorder that impacts a substantial number of people globally, exhibiting a chronic and enduring nature. A person with diabetes mellitus has excessive blood sugar due to either insufficient insulin production by the pancreas or insufficient cell response to the generated insulin. Diabetes mellitus is a collection of metabolic illnesses¹.

The global prevalence of diabetes is seeing a significant surge. A recent study reveals that the number of individuals affected by the condition increased from 171 million in 2000 to 422 million in 2014, showing a huge rise in the patient population. According to the most recent data from the World Health Organisation (WHO), the occurrence of diabetes mellitus has risen to 4.7%. Based on WHO projections, diabetes is expected to become the seventh most prevalent cause of death worldwide by 2030². Diabetic retinopathy is a prevalent disorder that impacts the small blood vessels of the retina and is frequently observed in individuals with diabetes. Diabetic ocular illness refers to the pathological state characterised by retinal impairment resulting from the presence of diabetes mellitus³. Abnormal ventricular functioning is a notable hallmark of diabetic cardiomyopathy, which arises as an extra consequence of diabetes mellitus. The most often given medicine for type 1 diabetes is insulin. In chronic situations, medication is also considered for type 2 diabetes and for the treatment of gestational diabetes mellitus⁴. Recently, several therapeutic techniques have been identified for the management of diabetes and its associated ramifications. The current formulation procedures employed do not sufficiently address the challenges pertaining to the delivery of anti-diabetic medicines. The main problem is that the medication candidates aren't very bioavailable when taken by mouth and don't stay stable in the gastrointestinal tract (GIT) because of the GIT's difficult conditions and low water solubility. Various nanoformulation techniques are being developed to tackle the issues related to oral bioavailability. Polymeric nanoparticles, self-nanoemulsifying drug delivery systems, nanoemulsions, solid lipid nanoparticles, nanostructured lipid carriers, liposomes, nanocrystals, and nanosuspensions are some of these methods^5-7. This article presents a comprehensive examination of the fundamental components of liposomal drug delivery systems. A concentric series of bilayers of phospholipids, either uni-lamellar or multilamellar, surround the internal aqueous compartment of liposomes, which are drug vesicles that self-assemble⁸.

A

B

Figure 1.A. Representative structure of a liposome. A lipid bilayer with an aqueous core is formed by the self-assembly of cholesterol and phospholipids. B. The most prevalent liposome subtypes are categorized⁹.

About forty years ago, Bangham and collaborators defined liposomes as circular vesicles that could be loaded with long-chain unsaturated fats, sphingolipids, glycolipids, non-toxic surfactants, and cholesterol¹⁰.

Liposomes vary in size from 30 nm to micrometers, whereas the phospholipid bilayer has a thickness of 4-5 nm. Liposome nanotechnology are often employed nanoparticles in nanomedicine because of their excellent drug loading efficiency^11-12, their biocompatibility stability, ease of synthesis, high bioavailability, as well as the secure excipients included in these formulation. Numerous physicochemical parameters, including vesicle size, surface charge, bilayer composition, coating, administration route, adjuvant use, encapsulation efficiency, and ultimately the lipid composition employed, influence the performance of liposome formulation¹³. At these delivery sites, liposomes can selectively and prolong their pharmacological effects by positively modifying the encapsulated species' pharmacokinetic profile¹⁴. In the pharmaceutical and cosmetic sectors, liposomes are frequently employed as carriers for an enormous variety of compounds¹⁵. As a result, a great deal of research has been done on liposomes to reduce medication toxicity and target particular cells for increased efficacy and safety. In order to achieve its pharmacological activity, the liposome merged with the cellular lipids membrane during the delivery of lipid materials, releasing liposomal content into the cell’s cytoplasm (Fig. 2). The several liposomal strategies using the hypoglycemic medications were shown in Table 1 of the article.

Consequently, it was discovered that liposomal vesicles were a useful tool for peptide, insulin, and GLP-1 distribution, and also for the release of water-soluble OHA for better control over the hyperglycemic phase of diabetes.

Anti-diabetic drug                Cell membrane

Fig. 2 - The way in which anti-diabetic drugs are included into liposomal formulation: medicines entering cells through the liposomal lipid layer’s infiltration on the lipid layer of the cell membrane¹⁶.

Table 1: Overview of anti-diabetic medication delivery systems based on liposomes.

Sr. No.	Type of DM (1/2)	Polymer	Size Range	Medication included	Outcome	Source
1	T1DM	Liposomes with biotin modification	160 nm	Insulin	It has been demonstrated that oral biotin (BLPs) absorption was aided via biotin receptor-mediated endocytosis.	16
2	T2DM	Micro complexation of Glycerolphosphate with Chitosan.	-	Metformin	With the suggested microcomplexes, a 40% increase in the AUC/D value and a 2.5-times longer Tmax of metformin were noted. The highly water-soluble antihyperglycemic medication is effectively transported via GP/CH microcomplexes, enabling regulated delivery and enhanced oral availability.	17
3	T1DM	Glycocholate sodium liposomes	150 nm	Insulin	Due to bile salts' ability to shield liposomal rhINS from enzymatic breakdown, sodium glycolate liposomes may present a viable option for protein and peptide delivery by mouth.	18
4	T1DM	Liposomes covered with chitosan	-	Insulin	Chitosan-coated liposomes that may improve insulin absorption via the GT tract by reducing degradation caused by enzymes.	19
5	T1/T2DM	Liposomes that contain deoxycholate of sodium, SGC, sodium taurocholate	400 nm	Insulin	SGC had the highest oral bioavailability and the hypoglycemic impact, followed by sodium taurocholate and sodium deoxycholate.	20
6	T1/T2DM	Lipid compositions within liposomes	(203.5 ± 20.5) nm	Insulin	The best ethanol percentage to attain the highest insulin encapsulation efficiency was discovered to be 40%.	21
7	T2DM	DPPC , DSPE-PG8G , Choles-terol are present in anionic liposomes.	131 nm	Glucagon-like peptide-1	When ionic liposomal administration was used with the composition specified in column "Polymer, " experimental rats showed a discernible improvement in the therapeutic benefits.	22

2. Liposome content:

2.1. Liposome-containing lipids:

Liposomes are spherical vesicles composed of lipid bilayers or diacyl-chain phospholipids that undergo self-assembly in aqueous solutions²³. Liposomes' electrical charge, rigidity, flow, stability, and size are all significantly influenced by the type of lipids that make up the particles^24,25. A acyl chain's length, symmetry, as well as saturation inside the hydrophobic group vary amongst lipids²⁶. The lipids that are used for producing liposomes fall under each of the following categories:

2.2. Natural lipid:

Two often encountered sources of natural phospholipids are egg yolks and soy beans²⁷. The stability of liposomes is influenced by the unsaturated character of the hydrocarbon chain, which renders natural lipids less stable compared to synthesized phospholipids^28,29.

2.3. Synthetic lipid

Synthetic phospholipids are generated through the application of specific chemical modifications to natural phospholipids. The change enables the creation of an extensive array of precisely defined and categorized phospholipids³⁰. The synthetic and saturated phospholipids are illustrated in (Figure 4)³¹.

2.4. Steroid:

Cholesterol is some other strategic aspect of liposomes. It affects the liposomes' lipid bilayer's modulatory characteristics³². Cholesterol is a steroid that is added throughout the liposome-making process to help strengthen their stiffness and stability³³. Phospholipid membranes can contain very high concentrations of cholesterol. It seems that cholesterol decreases blood protein interaction.

2.5. Surfactant:

Surfactants are molecules consist of a hydrophilic head and a lipophilic tail. The liposome capacity to encapsulate was altered when surfactants have been utilized to lower the surface tension inside different immiscible phases³⁴. Alkanes, aromatics, fluorocarbons, along with various non-polar groups can all function as surfactants. Liposomes primarily consist of non-ionic surfactants because of their low toxicity and biocompatibility. Surfactant induce the lipid bilayer of liposomal particles to rupture because they consist of acyl-chain amphiphile. (Figure 6)^35,36. Liposomes, which are composed of various surfactants, have been utilized extensively as drug delivery systems to enhance medication penetration³⁷.

3. Techniques for adding anti-diabetic medications and producing liposomes:

There is a wide range of ways available for producing liposomes, such as size reduction procedures and liposomal formulation techniques. Conventional and novel liposomal formulation procedures are the two types available.

3.1.  Conventional techniques:

The most often used conventional techniques for preparing liposomes include solvent injection, thin film hydration, reverse phase evaporation, and detergent removal, although there is a wide range of methods available³⁸.

3.1.1. Thin film hydration

Figure 3. Thin-film hydrate extraction technique.

A thin-film hydration process was the initial production technique for liposome development³⁹. In this procedure, a round-bottom flask is used to dissolve all of the lipids and the anti diabetic medication in an appropriate organic solvent. After that, the organic solvent gradually evaporated at lower pressure to produce a thin film layer. The resultant thin film is hydrated using an aqueous buffer solution. The main disadvantages of this process are that it produces more heterogeneous and larger liposomes, has a low entrapment ability, and is challenging to scale up and remove the organic solvent completely (Figure 7)^40.

3.1.2. Reverse-phase evaporation technique

Reverse-phase evaporation is used to produce an oil-and-water emulsion⁴¹. Organic solvent is evaporated at low pressure to produce a lipidic layer. After that, the organic solvent is eliminated, and nitrogen is continuously added to the system. Extrusion can decrease the polydispersity and average size of the liposome⁴². One advantage of this technology is its ability to achieve a greater level of encapsulation efficiency (EE)⁴³.

3.1.3. Solvent injection techniques

Figure 4. Diagrammatic illustration of injection technique.

In this method, lipids that have been dissolved in an organic solvent (like ethanol or ether) are quickly injected into an aqueous solution to make liposomes⁴⁴. The ethanol injection method is frequently employed in the production of liposomes due to its lack of oxidative alterations or lipid breakdown. Additionally, it is straightforward, replicable, quick to implement, and easily expandable⁴⁵. Moreover, it has been observed that extended exposure to elevated temperatures and organic solvents can potentially lead to a reduction in the stability of pharmaceuticals and lipids (see Figure 8)⁴⁶.

Phospholipids must dissolve in detergents at a sufficient concentration of micelles in order to complete the detergent removal process⁴⁷. A thin layer formed inside the flask as a result of the solvent evaporation⁴⁸. Following that, the lipid layer was hydrated in a water solution that contained drug molecules. This created a liposome solution⁴⁹. The next steps involve dialysis, size-exclusion chromatography, adsorption and dilution in order to eliminate surfactants^50,51,52,53. The ultimate liposomal formulation may harbour impurities; there is a possibility of a chemical reaction occurring between the detergent and the encapsulated substance, and this process is time-consuming⁵⁴.

3.2.  Techniques for reducing size:

Previous methods for liposome production necessitated additional procedures such as homogenization, extrusion, or sonication to reduce their size⁵⁵. Two separate sonication procedures, namely bath sonication and probe sonication, can be employed to control the size of liposomes. One possible limitation of the sonication technique is the difficulty in delivering an equivalent amount of ultrasonic energy within a large volume of liposomal suspension. Additionally, questions arise regarding low energy efficiency (EE), the potential degradation of phospholipids, and the medicine being encapsulated^56,57. Homogenization approaches involve the utilisation of a high-velocity collision idea, whereby liposomes are compelled to traverse an aperture under elevated pressure with the objective of reducing their size. The liposomes can be reduced in size using the extrusion technique. Following their formation, the liposomes are subjected to several extrusion cycles across a membrane that possesses a pre-established pore size, commonly a polycarbonate filter, in order to attain a uniform size distribution⁵⁸.

3.3.  Novel techniques:

Novel liposome preparation techniques are being researched primarily to enable industrial production scaling up and to be used with a variety of phospholipids and medications. There is innovative techniques resulting from the enhancement or adjustment of traditional approaches, such as the cross-flow injection (Wagner) technique⁵⁹, along with membrane contractor technology, modified ethanol injection techniques^60,61. The cross-flow filtration technique is designed using the improved detergent removal approach⁶². When liposomes are dissolved in water as co-solvent systems, they generate lipids and water-soluble carrier materials that, when freeze-dried, form cakes of an isotropic single-phase solution. A spontaneously homogenous dispersion of MLVs is formed when water is added to the freeze-dried product⁶³. Additionally, the utilisation of supercritical fluid (SCF) techniques has been investigated in the synthesis of liposomes. Several benefits of the SCF approach include the ability to produce on a large scale, the use of an inexpensive, environmentally safe solvent, the ability to adjust particle size, and in situ sterilizing. Lately, additional techniques, like dual asymmetric centrifugation as well as microfluidics, have also been used to create liposomes⁶⁴. The medicinal and pharmacological applications of all the innovative techniques mentioned above have great potential.

3.4.  Techniques for loading drugs:

Liposomes can encapsulate a wide variety of pharmaceuticals, they are thought to be an incredibly effective way to deliver medication⁶⁵. Drugs can be entrapped into liposomes using either of two techniques: passive or active. The passive loading method is the term used to describe the process of encapsulating medicine during the formation of liposomes. Several factors can influence the efficacy of drugs encapsulated through passive loading, including drug solubility, liposome size and charge, lipid concentration, and the manufacturing process⁶⁶. This approach frequently results in suboptimal encapsulation efficiency (EE), characterised by a substantial amount of unencapsulated drug and significant drug leakage for drugs that can pass through the liposomal bilayer⁶⁷. The procedure of active loading, alternatively referred to as distant loading, involves the efficient transportation of medication across the lipid bilayer through the creation of a transmembrane pH or ion gradient. In specific instances, this can lead to a loading rate of up to 100%. Following the synthesis of liposomes, this methodology is employed. There are several methodologies for conducting active loading, such as the phosphate gradient technique, the ethylenediaminetetraacetic acid (EDTA) gradient technique, the calcium acetate gradient technique for weakly acidic medications, the ammonium sulphate transmembrane gradient technique for amphipathic weak bases, and the ionophore loading approach⁶⁷.

4. Liposome Characterization:

Liposomes have several physiochemical properties, as listed in Table 2. These properties includes size distribution, the surface charge, structural, lamellarity, entrapped efficacy, polymorphism along with an in vitro dissolution profile.

Table 2: Describe the various techniques that are utilized to evaluate the parameters of liposomes.

Sr. No.	Liposomes characteristics	Characterization techniques	References
1	Particle size on average	Microscope technologies, including atomic force microscopy (AFM), cryogenic-TEM (Cryo-TEM), and Transmission electron microscopy (TEM), are combined with dynamic light scattering (DLS).	68,69
2	Surface charge and zeta potential	DLS and electrophoretic mobility	70
3	Particle morphology and shape	TEM, AFM, and Cryo-TEM	71
4	Lamellarity	Cryo-TEM	71
5	Phase behavior	Thermogravimetric analysis ,  X-ray diffraction and Differential scanning calorimetry	72,73
6	Drug release/encapsulation efficiency	centrifugation and dialysis, followed by chromatographic as well as spectrophotometric techniques to determine the amount of drug.	74,75

5. Liposome limitations:

For drug delivery applications, liposome stability is an important factor to take into account. Indeed, the duration and distribution of the medications within the liposomes within the body determines their therapeutic impact and safety, and these characteristics are closely linked to their stability. Liposomes must remain stable for a minimum of two years in order to qualify as a liposomal drug product, as per the Food and Drug Administration (FDA). The chemical and physical stability of liposomes are important elements that influence their biological efficacy⁷⁶. Many researchers have recently examined the adverse effects of liposomal doxorubicin, which is administered as a DOX using PEG-coated liposomes⁷⁷. The two main methods to assess liposomal physical stability are size measurement and visual examination of liposome appearance. Chemical stability refers to the ability of liposomes to maintain the amount of EE in the presence of oxidizing agents, pH changes, electrolyte composition, and surface active chemical. Changes in lipid membrane permeability may be caused by chemical degradation. Because therapeutic liposomal formulations are parenteral products that need to be sterilized in order to exclude microbiological contaminants from the finished product, the regulation of liposomal formulations’ microbial stability is particularly crucial.

6. CONCLUSION:

Many researchers have developed and investigated the possibilities of liposomes in drug delivery systems during the past few decades. The swift progress of liposomes presents alternative approaches to address limitations in the conventional delivery of anti-diabetic drugs, ultimately improving the management of individuals with diabetes. Liposomes have been shown to exhibit enhanced bioavailability, improved dose proportionality, reduced toxicity, and decreased dosing frequencies, hence indicating their superior efficacy in drug delivery. The investigation of liposomes' safety for extended use in enhancing drug delivery is currently underway, given the acknowledged importance of diabetes control through continuous monitoring of blood glucose levels and insulin. In order to convert innovation from distinct laboratory items into commercially feasible medical goods, it is imperative for professionals from diverse fields to engage in close collaboration. Therefore, the utilisation of liposomes in drug delivery systems is now in its nascent phase.

7. DISCLOSURES:

Considering the manuscript, there are no conflicts of interest or disclosures.

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Received on 26.03.2024      Revised on 09.07.2024

Accepted on 17.10.2024      Published on 18.04.2025

Available online from April 22, 2025

Asian J. Res. Pharm. Sci. 2025; 15(2):215-222.

DOI: 10.52711/2231-5659.2025.00033

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