TRANSFORMING THERAPEUTICS WITH STEALTH LIPOSOMES

TRANSFORMING THERAPEUTICS WITH STEALTH LIPOSOMES

Sukhmanpreet Singh *, Puja Gulati and Sachin Saggar

Desh Bhagat University, Fatehgarh Sahib, Punjab, India.

ABSTRACT: This review discusses PEGylated liposomes, also known as stealth liposomes, as modern nanocarriers that overcome the limitations of traditional drug delivery systems and pave the way for precision therapeutics. The article highlights that PEGylation gives stealth properties that include a significant increase in blood circulation time, greater stability, reduced immunogenicity, and improved passive tumor accumulation due to better permeability and retention. It also allows to achieve active targeting through ligand conjugation. The main sections cover the composition and mechanism of PEGylated liposomes, and their successful applications in cancer treatment, antimicrobial therapy, vaccines and immunotherapy, gene therapy, fungal infections, and autoimmune diseases. The review also discusses novel advancements, such as RNA loaded liposomes, while acknowledging critical challenges like accelerated blood clearance. In conclusion, despite facing significant challenges, PEGylated liposomes remain a key element of nanomedicine. Continued improvement in formulation, targeting strategies, and manufacturability will expand the impact beyond oncology to infectious, autoimmune, and genetic diseases.

Keywords: Liposomes, Pegylated Liposomes, Stealth Liposomes, PEGylation, Targeted drug delivery, Controlled drug release, Drug delivery systems

INTRODUCTION: The drug delivery systems are the technologies that provide the medicine to the specific sites in the body at a given time. It makes the medicine more effective and safer by increasing bioavailability, reducing the side effects, and allowing the release of the drug either in a sustained or targeted way. Different platforms like liposomes, micelles, nanoparticles, and hydrogels have been created to get rid of the disadvantages of conventional therapies and to facilitate chronic disease management and patient compliance, which in turn helps personalized medicine ¹. Among these, the major interest is gained by PEGylated liposomes.

By combining the liposomes with polyethylene glycol (PEG), they provide advantages like improved pharmacokinetics, stability, and superior therapeutic effects. These features make them widely employable in cancer and antimicrobial therapy, establishing them as a versatile platform in precision medicine ². Liposomes are spherical vesicles consisting of lipid bilayers that can encapsulate both hydrophilic and hydrophobic drugs, protecting them from degradation and thus enabling their controlled release ³. PEGylation provides a "stealth" feature, which decreases the immune clearance of liposomes ⁴.

This modification extends the circulation time considerably by improving the stability, reducing immunogenicity, preventing aggregation and increasing drug accumulation at disease sites due to reduced clearance ^{5, 6, 7}. This review focuses on PEGylated liposomes in terms of their structure, delivery mechanisms, key advances in pharmacokinetics and targeting, existing limitations, and future outlook.

Advantages of PEGylated Liposomes: PEGylation makes liposomes stay in the circulation for a longer time, going from minutes to hours or even days, thus enabling prolonged release of the drug and better therapeutic effect ⁸. It promotes passive targeting through the Enhanced Permeability and Retention (EPR) effect and can also allow active targeting if ligands such as antibodies or peptides are attached to the PEG chains ⁹. Lowered immune recognition opens the door for repeated dosing, while increased stability prevents drug leakage to an extent and facilitates controlled release ^{10, 11}. Accumulating drugs at the sites of disease decrease their systemic toxicity, which is a key factor in cancer treatment ¹². Consequently, PEGylated liposomes are able to provide more efficient and safer drug delivery, especially in the case of cancer therapy ¹³.

Composition of PEGylated Liposomes: PEGylated liposomes are made up of phospholipids, cholesterol that enhances the strength of the layer and protects against leakage, and PEG-lipid conjugates, which are responsible for their stealth properties ². The latter consist of amphiphilic molecules with covalently attached lipid anchors, such as Di stearoyl phosphatidylethanolamine (DSPE), which allow them to be securely embedded in the liposome bilayer ⁴⁸. The circulation and targeting ability are determined by the length and density of the PEG chains; greater PEG density leads to longer circulation, whereas lower density allows surface modification for active targeting ².

FIG. 1: REPRESENTATION OF PEGYLATED LIPOSOME STRUCTURE

Mechanism of Action: PEGylated liposomes are capable of drug delivery through both passive and active targeting mechanisms.

Enhanced Permeability and Retention (EPR) Effect: Due to the permeable nature of the tumor's blood vessels, the EPR effect takes place in tumors, which are the main sites for the accumulation of PEGylated liposomes besides the blood circulation ¹⁴. The main contributors are as follows:

Abnormal Blood Vessels: Abnormalities in tumor blood vessels such as large gaps in endothelial cells and immature vascular structures increase the permeability for nanoparticles and thus, PEGylated liposomes and other larger molecules can diffuse from the blood into the tumor extracellular space easily ¹⁵. 

Poor Lymphatic Drainage: The lymphatic drainage system in normal tissues is very effective in getting rid of interstitial fluid and large molecules. In contrast, ineffective lymphatic drain in tumors will retain liposomes longer and hence, increase the drug concentration and therapeutic effect in the tumor area ¹⁶.

FIG. 2: ENHANCED PERMEABILITY AND RETENTION (EPR) EFFECT

Active Targeting: Active targeting alters the liposomal surface through the use of ligands that bind to receptors highly present on cancer cells thereby enhancing selectivity and cellular uptake ¹⁷. Antibodies, peptides, and aptamers significantly increase the interaction with the tumor tissue.

Some examples are:

• Folate-conjugated PEGylated liposomes, which at the same time enhance delivery and are selectively taken by ovarian cancer cells with folate-receptor overexpression ¹⁸.
• Liposomes modified with transferrin have shown to be more efficient in drug uptake in the cancer cells due to receptors overexpression ⁹.

These methods facilitate precision targeting beyond passive EPR accumulation.

FIG. 3: ACTIVE TARGETING OF LIPOSOMES

Applications of PEGylated Liposomes:

Cancer Therapy: The delivery of cytotoxic chemotherapy agents, such as doxorubicin, has been greatly altered by PEGylated liposomes and, hence, cancer therapy has immensely benefitted from PEGylated liposomes. Doxil, the PEGylated liposomal drug that was the first to be approved by the U.S. Food and Drug Administration (FDA), showed a significant decrease in cardiotoxicity as compared to free doxorubicin ¹⁹. Likewise, other drugs like irinotecan and paclitaxel have also shown better efficacy when formulated in PEGylated liposomes. The latest studies also seem to back this approach. For instance, the usage of PEGylated liposomes for palbociclib in triple-negative breast cancer allowed for a 2.5-fold increase in the drug level in the cancer area as well as a decrease in the side effects caused by the drug when compared to the free drug, all leading to better therapeutic results ²⁰. The liposomal formulation of 5-fluorouracil co-evaluated for increased pharmacokinetics and decreased cardiotoxicity, exhibiting the drug’s greater effectiveness in animal tumor models ⁴⁶.

Antimicrobial Therapy: The encapsulation of antibiotics like vancomycin and amphotericin B in PEGylated lipids improved their therapeutic index by concentrating on infected tissues and reducing off-target effects ^{21, 22}. Moreover, in new studies, targeted PEGylated liposomes with ciprofloxacin showed better drug retention and higher survival rates in Pseudomonas aeruginosa-infected mice ²³. Also, it was shown that PEGylated liposomes with endolysins were successful in disrupting the bacterial biofilms formed by Staphylococcus pneumoniae ⁴⁷.

Vaccines and Immunotherapy: PEGylated liposomes are utilized as antigen and immune adjuvant carriers to increase immunogenicity and at the same time, reduce systemic toxicity. Such systems have been tested for the treatment of diseases like influenza and cancer ²⁴. Recent studies reveal that PEGylated cationic liposome vaccines have better antigen drainage to lymph nodes and induce strong immune responses against influenza in murine models ²⁵.

Moreover, liposomal delivery of tumor-associated antigens resulted in T-cell activation and reduced systemic toxicity in cancer immunotherapy ²⁶. Research findings further show that PEGylation in liposomal adjuvant systems enhances vaccine efficacy by promoting dendritic cell uptake and antigen presentation ²⁷.

Gene Therapy: The diversity of PEGylated liposomes is also applied to the delivery of genetic material, for example, plasmid DNA or interfering RNA (siRNA). PEGylation plays a significant role in the sustaining of nucleic acids' stability in circulation and, consequently, their delivery to the exact sites of action ²⁸. Numerous studies pointed out the benefit of use of such liposomes with folic acid conjugated PEGylated liposomes that deliver siRNA with cancer cell selectivity, thus increasing the uptake of the siRNA and the silencing of the gene both in-vitro and in-vivo ⁴⁵. PEGylated liposomes with plasmid DNA showed better circulation and better gene expression in the target tissues. The PEGylated liposomal system came up with a longer circulation which could be helpful in the gene-based therapy of diseases ²⁹.

Fungal Infections: Liposomal amphotericin B (AmBisome®) has been very successful in the treatment of systemic fungal infections and at the same time has been less nephrotoxic than the traditional formulations. Moreover, liposomal amphotericin B was favored over the conventional treatment of invasive fungal infections in terms of good safety and effectiveness ²¹. Latest researches indicate that PEGylated liposomes containing Ciclopirox olamine led to a significant reduction in fungal burden, which was accompanied by no toxicity in the immunosuppressed mice that had been infected with Cryptococcus neoformans ³⁰. Further research showed that PEGylated liposomal formulations also enhanced antifungal activity against Candida auris ³¹.

Autoimmune Diseases: Dexamethasone and other anti-inflammatory agents in liposome formulations have proven to be efficacious in the treatment of rheumatoid arthritis ³². It has been demonstrated that PEGylated liposomal formulations of such anti-inflammatory agents can both lower toxicity and increase the effectiveness of treatment in models of autoimmune diseases ³³.

TABLE 1: KEY FINDINGS OF PEGYLATED LIPOSOMES APPLICATIONS

Emerging Innovations in Liposome Technology:

Stimuli Responsive Liposomes: Stimuli-responsive liposomes release their drug in response to external stimuli, e.g., pH, temperature, or light. For example, pH-sensitive PEGylated liposomes are made to release drugs in acidic tumor microenvironments to increase drug accumulation at the tumor site ²². In tumor environments, pH-sensitive and thermosensitive liposomal nanoformulations allow for highly selective drug release, increasing drug accumulation and therapeutic efficacy, according to recent research ³⁴. In models of hepatocellular carcinoma, multifunctional liposomes that combine phototherapy and gene-drug delivery have demonstrated potent tumor suppression when activated by external stimuli ³⁵.

Hybrid Liposomes: Hybrid liposomes merge lipids with polymers or inorganic nanoparticles for multifunctional use. PEGylated hybrid liposomes containing gold nanoparticles have shown theranostic capability by allowing drug delivery and imaging at the same time ¹¹. Drug delivery and real-time tracking in cancer treatments are made possible by core-shell hybrid liposomes that incorporate inorganic nanoparticles like gold. For the best theranostic results in tumor models, a different study showed hybrid liposome–gold nanostructures that enable protected chemotherapeutic delivery and bioimaging at the same time ³⁶.

Liposomal RNA Delivery: Emerging progress with RNA therapeutics, such as siRNA and messenger RNA (mRNA), has made PEGylated liposomes an attractive delivery system. mRNA-based COVID-19 vaccines, for example, by Moderna and Pfizer, delivered via lipid nanoparticles have shown the flexibility of this system to tackle public health issues ²⁸.

In ovarian cancer models, folate-modified PEGylated liposomes have demonstrated efficacy in delivering CRISPR plasmid DNA, resulting in potent anti-tumor effects and improved gene transfer ³⁷. The stabilization of siRNA and mRNA is emphasized in reviews of PEGylated lipid-based RNA delivery systems ³⁸.

Immunoliposomes: Immunoliposomes, one of the target liposomes, are designed through the conjugation of antibodies or antibody fragments to the surface of liposomes. These liposomes have increased affinity for particular antigens. As an example, trastuzumab-conjugated liposomes target HER2-positive (human epidermal growth factor receptor-2) carcinoma cells with exceptional specificity, bringing about enhanced therapeutic efficacy in treating breast cancer ². In cervical cancer-on-a-chip experiments, Anti-programmed death-ligand1 (PD-L1) immunoliposomes demonstrated enhanced drug delivery and cancer cell targeting, proving their usefulness for cancer immunotherapy ³⁹. In lymphoma models, valrubicin-loaded immunoliposomes with dual targeting markers markedly increased immunological modulation and antitumor efficacy ⁴⁰. However, PEGylation can interfere with antibody binding due to steric hindrance. Researchers are exploring optimized PEG chain lengths and densities to balance circulation time and targeting efficacy ²⁴. Immunoliposomes represent a promising innovation for treating cancers and autoimmune diseases where specific targeting is essential.

TABLE 2: KEY FINDINGS OF EMERGING INNOVATIONS

Challenges in PEGylated Liposomes: Despite the therapeutic benefits, some challenges are encountered in the development of PEGylated liposomes. One of the main hindrances is the occurrence of the ABC phenomenon during the recurring use of PEGylated liposomes, which is caused by the production of anti-PEG antibodies, finally resulting in reduced circulation time and thus limiting the effectiveness of long-term treatments ⁴¹. The manufacturing process has also remained very complicated, as there is a need for an exact control over the lipid composition, PEG density, and particle size in order to achieve consistency from one batch to another ⁴². In addition, large-scale production involves not only the high-purity lipids and PEG derivatives but also the specialized processing equipment, which raises the price and thus limits the availability of the product ⁴³. Apart from these issues, storage instability has also been a challenge as the lipids in PEGylated liposomes. They can undergo oxidation and hydrolysis, which in turn, causes them to either destabilize or degrade. Freeze-drying with an appropriate cryoprotectant is commonly employed to enhance the long-term stability ^{44, 19}.

CONCLUSION: PEGylated liposomes have proven to be one of the efficient drug-delivery platforms by improving the circulation time and stability, as well as targeted accumulation, with minimum systemic toxicity. Its successful application in oncology, antimicrobial therapy, vaccines, gene delivery, and autoimmune diseases demonstrates strong translational and clinical relevance supported by approved formulations.

However, accelerated blood clearance and manufacturing complexity have resulted in continuous formulation optimization to address such limitations. Promising strategies, including stimuli-responsive system hybrid liposome, RNA delivery platform, and immunoliposomes, are foreseen to further improve targeting precision and therapeutic performances.

With continued advancements, PEGylated liposomes will play a crucial role in advanced nanomedicine by enabling safer and more effective treatment strategies in future clinical applications.

ACKNOWLEDGEMENTS: The authors would like to express their gratitude to all who supported and provided their guidance and encouragement throughout the preparation of this review.

CONFLICTOF INTEREST: The authors declare that there is no conflict of interest regarding the publication of this article.

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    How to cite this article:

    Singh S, Gulati P and Saggar S: Transforming therapeutics with stealth liposomes. Int J Pharm Sci & Res 2026; 17(5): 1455-61. doi: 10.13040/IJPSR.0975-8232.17(5).1455-61.

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