Flexible Liposomes Research Articles

Intratumoral lactic acid neutralization strategy for boosting chemoimmunotherapy using liposomal sodium bicarbonate

Glycolysis-related lactic acid overproduction creates an “ion-trapping” barrier and immunosuppressive tumor microenvironment that compromise effective intratumoral drug delivery and therapy. Therefore, normalization of tumor microenvironment via lactic acid neutralization can be a promising avenue for overcoming this therapeutic hurdle. In this study, the flexible liposomes loaded with sodium bicarbonate (NaHCO3@Flip) were used as a nano-adjuvant to boost chemoimmunotherapy. Their effects on assisting DOXIL and anti-PD-1 therapy were investigated. NaHCO3@Flip achieved deep tumor penetration, with the ability to neutralize lactic acid and normalize the acidic tumor microenvironment. NaHCO3@Flip is biosafe and can enhance cellular uptake efficiency of doxorubicin (DOX) by overcoming the ion-trapping barrier and amplify immunogenic cell death induced by DOX. The combination therapy of liposomal DOX and NaHCO3@Flip demonstrated enhanced inhibition of tumor growth. NaHCO3@Flip can also synergize with PD-1 antibody therapy. NaHCO3@Flip has the potential to serve as a therapeutic adjuvant for boosting chemoimmunotherapy by overcoming the ion-trapping effect and normalizing the tumor microenvironment.

Alpha-Tocopherol-Infused Flexible Liposomal Nanocomposite Pressure-Sensitive Adhesive: Enhancing Skin Permeation of Retinaldehyde

Retinaldehyde (RAL), or retinal, is a vitamin A derivative that is widely used for several skin conditions. However, it is light sensitive and has low water solubility, limiting its efficiency in transdermal delivery. This study developed a novel delivery system for retinal (RAL) using flexible liposomes (FLPs) infused with α-tocopherol succinate (α-TS) to improve stability, and enhance skin permeability. The RAL-FLPs were embedded in pressure-sensitive adhesive (PSA) hydrogels, creating a delivery platform that supports prolonged skin residence and efficient permeation of RAL. The stability and skin permeation as well as human skin irritation and adhesion capabilities were assessed to determine the formulation’s safety and efficacy. Our findings suggested that the addition of α-TS could improve liposomal stability and RAL chemical stability. Moreover, the skin permeation and fluorescence microscopic-based studies suggested that the addition of α-TS could enhance skin permeability of RAL through hair follicles. The RAL-FLP was embedded in PSA hydrogels fabricated from 25% GantrezTM S-97 (GT) and 1% hyaluronic acid (Hya) with aluminum as a crosslinker. The PSA hydrogel exhibited desirable peeling and tacking strengths. The developed hydrogels also demonstrated greater skin deposition of RAL compared with its aqueous formulation. Additionally, the RAL-FLP-embedded PSA hydrogels showed no skin irritation and maintained better adhesion for up to 24 h compared to commercial patches. Hence, the developed hydrogels could serve as a beneficial platform for delivering RAL in treating skin conditions.

In vivo pharmacokinetic study of carmustine in rats after giving single-dose of carmustine API solution, flexible liposomes, in situ nasal gel, optimized flexible liposomes embedded in situ nasal gel, and marketed formulation

Carmustine is used in the treatment of glioblastoma (GBM). GBM is a well-known life-threatening type of cancerous tumor. GBM covers 60.00% among all primary brain tumors, with an occurrence of 74,000 cases across the globe. Management for GBM is still very difficult because most of the medicines are unable to cross the blood-brain barrier (BBB). The present work observed that flexible liposomes embedded in situ nasal gel of carmustine is the best brain-targeted medicine delivery system for the management of GBM through the nasal route. To evaluate in vivo pharmacokinetic parameters of carmustine formulations administered through nasal routes in Wistar rats. In this work, different pharmacokinetic parameters were determined for carmustine formulations viz. carmustine API (Active Pharmaceutical Ingredient) solution, flexible liposomes, in situ thermoreversible intranasal gel, optimized flexible liposomes embedded in situ thermoreversible intranasal gel via intranasal administration in rats, and compared with marketed intravenous injection of carmustine administered through intravenous route. Carmustine was estimated with the help of a validated high-performance liquid chromatography (HPLC) approach. Three to four-months-old normal Wistar rats of either sex, having a weight of 200-250 grams were used in this study. Intranasal administration of optimized flexible liposomes embedded in situ nasal gel showed greater Cmax (∼two-fold), AUC0→t (∼three-fold), AUC0→∞ (∼six-fold), and decreased Tmax (1h) data in the brain, than commercial intravenous injection of carmustine. The plasma concentration of carmustine administered through nasal route was found to be comparatively lower than intravenous administration, indicating lower systemic exposure to carmustine via the nasal route. In vivo pharmacokinetics results revealed that the optimized flexible liposomes embedded in situ nasal gel of carmustine can effectively deliver carmustine to brain by nasal drug delivery system in Wistar rats.

Engineered Exosomes Biopotentiated Hydrogel Promote Hair Follicle Growth via Reprogramming the Perifollicular Microenvironment.

Androgenetic alopecia (AGA) is a highly prevalent condition in contemporary society. The conventional treatment of minoxidil tincture is hindered by issues such as skin irritation caused by ethanol, non-specific accumulation in hair follicles, and short retention due to its liquid form. Herein, we have developed a novel minoxidil-incorporated engineered exosomes biopotentiated hydrogel (Gel@MNs) that has the capability to modulate the perifollicular microenvironment for the treatment of AGA. Leveraging the exceptional skin penetration abilities of flexible liposomes and the targeting properties of exosomes, the encapsulated minoxidil can be effectively delivered to the hair follicles. In comparison to free minoxidil, Gel@MNs demonstrated accelerated hair regeneration in an AGA mouse model without causing significant skin irritation. This was evidenced by an increase in both the number and size of hair follicles within the dermal layer, enhanced capillary formation surrounding the follicles, and the regulation of the transition of hair follicle cells from the telogen phase to the anagen growth phase. Therefore, this safe and microenvironment-modifying hybrid exosome-embedded hydrogel shows promising potential for clinical treatment of AGA.

Brain Targeted Drug Delivery System of Carmustine: Design, Development, Characterization, in vitro, ex vivo Evaluation and in vivo Pharmacokinetic Study

The treatment of gliomas remains difficult task. Carmustine is a drug that is used in the treatment of gliomas. Flexible liposomes embedded in situ thermoreversible nasal gel preparations of Carmustine were studied for in vitro carmustine release, ex vivo carmustine permeation and carmustine release kinetics. The epithelial layers of nasal tissue were found to be intact and undamaged during histological analysis. Intranasal administration of optimized flexible liposomes embedded in a nasal gel showed higher Cmax (Approximately two-fold), AUC0→t (Approximately three-fold), AUC0→∞ (Approximately six-fold), and lower Tmax (1 h) in the brain, compared to intravenous injection of carmustine. The present study demonstrates that the flexible liposome embedded thermoreversible in situ intranasal gel of carmustine improved the targeted uptake of carmustine in the brain through the nasal delivery system and could be a reliable and effective delivery system for carmustine in the treatment of gliomas.

EVALUATION OF IN VITRO CYTOTOXIC POTENTIAL OF DIFFERENT CARMUSTINE FORMULATIONS AGAINST U-87 MG HUMAN GLIOBLASTOMA CELL LINE

Glioblastoma (GBM) is a frequent as well as violent type of brain tumor. In this research work, different carmustine formulations were prepared and screened for their cytotoxic activity against U-87 MG glioblastoma and normal human fibroblast L-929 cell lines. The flexible liposomes embedded in situ nasal gel exhibited maximum percentage of growth inhibition against U-87 MG glioblastoma cell line, and in situ nasal gel exhibited lowest percentage of growth inhibition against U-87 MG glioblastoma cell lines. Flexible liposomes embedded in situ nasal gel is observed to be safe and biocompatible against normal human fibroblast L-929 cell line. In cellular uptake study, U-87 MG cell line treated with flexible liposomes embedded in situ thermoreversible intranasal gel emitted stronger and higher intensity fluorescence. It seems that the presence of flexible liposomes embedded in situ thermoreversible nasal gel inside the tumor cells, and would be the best carmustine delivery approach for the management of GBM.

Evaluation of F3S4-m loaded liposomes as anti-AChE and its cytotoxic activity in PC12 and HMC3 cells

The current research discusses the loaded of F3S4-m compound reported with multitarget activity acting as an inhibitor of acetylcholinesterase (AChE), beta-secretase 1 (BACE1), and amyloid beta (Aβ) aggregation. Despite great effort has been done to have a compound to treat Alzheimer’s disease (AD) and not only its symptomatology, nowadays there is not an effective compound in the market yet. Several synthetized compounds reported having a good activity in vitro study turned out not having it in vivo, it is believed that many of them are not able to reach the brain due to low biodisponibility, besides many of them have charged chemicals groups or tertiary amines, which have been substrates of deaminases enzymes. Therefore, the use of liposomes results an interesting strategy to deliver this kind of compounds to the brain. Then in this work it is proposed the use flexible liposomes made from phosphatidylcholine (PC) and cholesterol (Chol) to encapsulate the F3S4-m compound and evaluate its inhibitory activity against AChE. The results showed that empty liposomes particle size is influenced by the content of cholesterol in their membranes being larger for those containing 20% of cholesterol than those with 10%. The toxicity of F3S4-m loaded liposomes in PC12 cells was also evaluated employing the 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide assay (MTT). The results showed that F3S4-m loaded liposomes can inhibit the enzymatic activity of AChE, but the concentration of encapsulated F3S4-m needed to inhibit AChE was 2-times higher than what was previously reported in solution.Graphical abstract

Trimethylated chitosan-coated flexible liposomes with resveratrol for topical drug delivery to reduce blue-light-induced retinal damage

LED-related blue-light-induced damage can cause eye diseases. However, drug delivery in patients with ocular diseases is faced with various challenges. In this study, we developed flexible liposomes based on trimethylated chitosan (TMC-Lipo) to deliver resveratrol for the treatment of retinal diseases. Flexible liposomes can easily cross various biological barriers. Chitosan and its derivatives have adhesive properties and are widely used in mucoadhesive drug delivery systems. Therefore, we wrapped flexible liposomes with trimethylated chitosan via electrostatic adsorption. The charge of the flexible liposomes became positive after encapsulation in TMC, and they remained stable in artificial tears. We assessed the safety of TMC-Lipo in cellular and zebrafish experiments and found that it can be safely used. In addition, treatment with TMC-Lipo significantly reduced H2O2-induced damage to ARPE-19 cells, restored mitochondrial membrane potential, and protected the cells. TMC-Lipo more easily reached the posterior ocular segment of the mice than liposome nanoparticles and attenuated blue-light-induced retinal cytopathy. Our study demonstrates that effective eye drop formulations can be developed based on trimethylated chitosan, which provides a promising approach for the treatment of ocular diseases.

Essential Oil from the Twigs of Cinnamomum Cassia Mediated the Flexible Liposomes of the Total Glucosides of Peony for Arthritis Treatment

Flexible liposomes (FL) have been developed as novel transdermal delivery systems. However, the skin barrier impairs the drug penetration and absorption. In this study, we designed, characterized and evaluated a total glucosides of Paeony (TPG) loaded FL (TFL) topically drug deliver system for arthritis treatment. The novelty is that the essential oil from the Twigs of Cinnamomum Cassia (EOCC) was incorporated in TFL as drug penetrating enhancer. Interestingly, the EOCC addition didn’t significantly change the characteristic parameters of the TFL in terms of morphology, size distribution, zeta potential, and entrapment efficiency. Remarkably, EOCC-embedded TGP-loaded FL (EOCC-TFL) dramatically prevented the drug leakage and improved the TGP skin penetration in vitro (4.6, 1.8, and 1.5 times higher than the TGP solution, TGP and EOCC mixture, and TFL, respectively). Moreover, despite of the low dose (175 mg/kg which was half dose of the TGP gavage group), this optimized formulation exhibited favorable anti-inflammatory effects in vivo on arthritis rats with significantly reduced swollen toe volume and serum inflammatory factor levels. This study lays solid foundation for the engineering liposomes with EOCC for transdermal drug delivery.

Enhanced Transdermal Peptide-Modified Flexible Liposomes for Efficient Percutaneous Delivery of Chrysomycin A to Treat Subcutaneous Melanoma and Intradermal MRSA Infection.

Superficial skin diseases, including skin infections and tumors, are common healthcare burdens. In this study, the in vivo activity of chrysomycin A (CA) is explored, and a transdermal liposomal CA formulation is further constructed for the simultaneous treatment of cutaneous melanoma and cutaneous methicillin-resistant Staphylococcus aureus (MRSA) infection. The prepared liposomes (TD-LP-CA) display a strong antitumor effect with an IC50 value of less than 0.1µm in B16-F10 cells, suppress the proliferation of MRSA with a minimum inhibitory concentration (MIC) of 1µm, and eradicate established MRSA biofilms at 10× MIC in vitro. More importantly, TD-LP-CA shows enhanced stratum corneum (SC) penetration, reaching more than 500µm beneath the skin's surface due to modification with the TD peptide, and demonstrates excellent subcutaneous tumor penetration after skin application in vivo. TD-LP-CA displays an excellent therapeutic effect against intradermal MRSA infection in mice after topical dermal administration, as well as a moderate inhibitory effect on subcutaneous melanoma with a 75% tumor inhibition rate. The liposomes prepared herein can be a promising carrier for transcutaneous CA transfer for the treatment of superficial diseases such as skin tumors and infections due to their ability to overcome the skin barrier.

Anti-photoaging effect and mechanism of flexible liposomes co-loaded with apigenin and doxycycline

Prolonged exposure to UV light can lead to photo-ageing of the skin. Therefore, the development and application of anti-photoaging drugs is urgent. In this study, we co-loaded apigenin (Apn) and doxycycline (Doc), a broad-spectrum inhibitor of matrix metalloproteinases (MMPs), into flexible liposomes to exert anti-photoaging effects by combating oxidative stress, anti-inflammation, reducing the activation of MMPs and preventing collagen loss. The results showed that we prepared a flexible liposome (A/D-FLip) containing Apn and Doc. Its appearance, particle size and Zeta potential were normal and it had good encapsulation efficiency, drug loading, in vitro release and transdermal efficiency. In cellular experiments, A/D-FLip could inhibit oxidative stress damage, reduce inflammatory factors and decrease the activation of MMPs in Human immortalized keratinocytes (HaCaT) cells; in animal experiments, A/D-FLip could inhibit skin damage and reduce skin collagen loss by decreasing the activation of MMPs, thus inhibiting skin photoaging in mice. In conclusion, A/D-FLip has good anti-photoaging effects and it has the potential to become an effective skin care product or drug against UV damage and skin photoaging in the future.

High skin permeation, deposition and whitening activity achieved by xanthan gum string vitamin c flexible liposomes for external application

For most external applied antioxidant whitening ingredients, effective stratum corneum breakthrough, epidermal penetration and dermal deposition are necessary premises for inhibition of melanin production and transfer occurring in stratum basale. Herein, xanthan gum was added into vitamin C-containing flexible liposome (Vc-L) suspension. The long polymer chain of xanthan gum string dispersed Vc-L together to gain a lotion (Vc-LX) for external application. In this study, the storage stability experiments demonstrated that the additional xanthan gum could improve the storage stability of Vc liposome suspension. The cumulative in vitro skin penetration and deposition of Vc-LX was found to significantly increase within 24 h in mouse skin, compared with those of the Vc aqueous solution and Vc conventional liposomes treated groups (***p < 0.001). Most importantly, in vivo skin whitening experiments gave that Vc-LX has better skin whitening activity (ΔL*) than marketed products (GARNIER® Vc377), Vc flexible liposomes, and Vc conventional liposomes. Moreover, in vitro cytotoxicity experiments and skin irritation experiments demonstrated that Vc-LX has good biosafety. Therefore, this study suggested that Vc-LX may be a promising local skin delivery system for water-soluble antioxidant ingredients.

Comparison of Therapeutic Effects of Topical Application of Diclofenac Sodium Nanoparticles and Conventional Placebo on Knee Osteoarthritis.

The objective of this study was to investigate the therapeutic effects of diclofenac sodium nanoparticles and traditional placebo on Knee Osteoarthritis (KOA). 80 KOA patients admitted to the Inpatient Department of Orthopedics of Sunshine Union Hospital from June 2020 to July 2021 were selected, and divided into 40 cases using diclofenac sodium nano flexible liposomes topically (study group) and 40 cases using conventional placebo topically (control group). The two groups were compared for knee swelling, knee pain, and knee function scores before treatment, on the 3rd, 7th, and 14th days after treatment, and the incidence of adverse events during treatment was recorded. It was found that the knee swelling scores of the study group were significantly lower than those of the control group on the 7th and 14th days after treatment, and that the knee pain score of the study group was significantly lower than that of the control group at the 7th day after treatment, and that the knee joint mobility disorder scores of the study group were significantly lower than those of the control group at the 7th and 14th days after treatment. The total effective rate of the study group was 85.0% (34/30), which was higher than that of the control group (12.5% (5/30)) (P<0.05). In conclusion, local application of diclofenac sodium nano flexible liposomes showed a stronger effect on KOA than traditional placebo, and no significant adverse reactions occurred.

Formulation and Evaluation of Flexible Liposome Embedded In Situ Thermoreversible Intranasal Gel of Rizatriptan Benzoate

The present study deals with formulation and evaluation of flexible liposome embedded thermoreversible in situ intranasal gel of rizatriptan benzoate. The flexible liposomes were formulated (F1-F9) and optimized with respect to percentage of ethanol and phospholipid to get optimum vesicle size and entrapment efficiency. The vesicles were characterized for their size, shape and zeta potential. The optimized stable vesicles were incorporated into in situ thermoreversible gel. The ex vivo permeation of drug through goat nasal mucosa was studied. The nasal mucosa used for drug permeation experiment was subjected to histopathological study to check for any damages. The in vivo pharmacokinetic study was carried out in Wistar rats. Vesicles size and entrapment efficiency of stable formulations (F2-F6) was varied from 71.2±14.16 to 199.3±26.03 nm and 72.98 % to 77.44 % respectively. The liposomes were nearly spherical in shape with zeta potential varied from -14.2±4.48 to -11.7±5.26 mV. The improved drug permeation and higher flux across nasal mucosa with enhancement ratio of 2.75 was observed for flexible liposomal thermoreversible gel F2 compared to normal thermoreversible hydrogel of drug. The histopathological study of nasal mucosa revealed that there was no damage to epithelial layer and was found to be intact. The area under the curve of optimized formulation (F2) was 3.98 and 1.53 times more than oral drug solution and intranasal gel respectively, confirming improvement in bioavailability. Thus present study concludes that flexible liposome enriched thermoreversible, mucoadhesive in situ intranasal gel of rizatriptan benzoate is a safe and effective drug delivery system for treatment of migraine.

Nanotherapeutics a promising approach for treatment of Parkinson’s disease

Parkinson's disease (PD) is a chronic, neurodegenerative disorder that progresses in extrapyramidal region. It affects about 5 million people worldwide, and the total cost of treatment is supposed to surpass $38.6 million if no effective treatments are produced. An alarming rise of 1.3 million cases is expected in forthcoming years.The fundamental goal of new treatment techniques for PD is to restore dopaminergic tone in the substantia nigra and striatum. The majority of conventional medicines follow the oral routes of administration, which have a low specificity towards brain. Effective methods of drug delivery to brain can improve the specificity, bioavailability, and patient compliance while reducing unwanted side effects. Blood brain barrier (BBB) is a major hurdle for conventional drugs as it doesn’t allow hydrophobic and large sized molecules to pass through and reach the brain. Nanotherapeutics can help to overcome the BBB, and nanoparticles have lately been widely employed to load different medicines, hormones, and phytochemicals for treating PD.To transfer the quantity of drugs of our choice to certain brain regions several nanocarriers such as nanoparticles (NPs), polymeric micelles, nanoemulsion, dendrimers, nanosuspension, solid lipid nanoparticles (SLNPs), flexible liposomes (transferosomes), carbon nanotubes, viral vectors, immunoliposomes, and antibodies and their conjugates have been developed. The current review gives an idea about various nanotherapeutics employed for the treatment of PD.

Recent Advances in Elastic Liposomes: Current Status and Future Perspective

Elastic liposomse (ELs) are the flexible liposomes formulated using phospholipids as well as edge activators. Edge activators provide elasticity to the Els. Els provide advantages over other formulations and have the ability to be delivered by different routes such as topical, transdermal, nasal, ocular, etc. Potential of encapsulating not only lipophillic but also hydrophillic drugs in a single vesicle, ability to pass through channels 1/10th of their diameter, increase in drug permeation, enhanced solubility of drug, patient compliance, prevention of degradation of drug makes them efficient carriers of drugs and leads to increased interest of researchers in them. This review provides understanding of composition of Els, advantages, method of preparation and the adaptable role played by ELs in the administration of numerous drugs for different diseases.

FtsZ induces membrane deformations via torsional stress upon GTP hydrolysis

FtsZ is a key component in bacterial cell division, being the primary protein of the presumably contractile Z ring. In vivo and in vitro, it shows two distinctive features that could so far, however, not be mechanistically linked: self-organization into directionally treadmilling vortices on solid supported membranes, and shape deformation of flexible liposomes. In cells, circumferential treadmilling of FtsZ was shown to recruit septum-building enzymes, but an active force production remains elusive. To gain mechanistic understanding of FtsZ dependent membrane deformations and constriction, we design an in vitro assay based on soft lipid tubes pulled from FtsZ decorated giant lipid vesicles (GUVs) by optical tweezers. FtsZ filaments actively transform these tubes into spring-like structures, where GTPase activity promotes spring compression. Operating the optical tweezers in lateral vibration mode and assigning spring constants to FtsZ coated tubes, the directional forces that FtsZ-YFP-mts rings exert upon GTP hydrolysis can be estimated to be in the pN range. They are sufficient to induce membrane budding with constricting necks on both, giant vesicles and E.coli cells devoid of their cell walls. We hypothesize that these forces result from torsional stress in a GTPase activity dependent manner.

Chitosan caged liposomes for improving oral bioavailability of rivaroxaban: in vitro and in vivo evaluation

In this study, chitosan (CS) caged classic liposomes (CLs) and flexible liposomes (FLs) were developed to enhance the oral bioavailability of rivaroxaban (RVX) in the fasted condition. The prepared formulations were subjected to physicochemical characterization included: FTIR, DSC, zeta potential, particle size, polydispersity index, entrapment efficiency, in vitro dissolution, and transmission electron microscope imaging. The selected formulation (RVX-TFL2) composed of PL S100/Tween 80 (85/15% w/w) and coated with CS solution in the strength of (0.2% w/v) had a particle size of 105.67 nm, a zeta potential of +5.67 mV and EE of 96.07%. Compared to RXV suspension, the pharmacokinetic parameters (C max, AUC0-24, and AUC0-∞) of RVX-TFL2 showed no statistically significant difference (P > 0.05) in the fasted and fed test animals. Besides, RVX bioavailability with RVX-TFL2 was improved by 59.66% and 26.97% in the fed and fasted states, respectively, compared to RVX suspension in the fed state. The result highlighted the efficacy of the prepared liquid formulation comprising CS coated liposomes in improving the oral bioavailability of RVX regardless of the fed state. Moreover, the studied liquid formulation could be utilized in developing a liquid dosage form that might be useful as a pediatric formulation of RVX.

Microwave hyperthermia-responsible flexible liposomal gel as a novel transdermal delivery of methotrexate for enhanced rheumatoid arthritis therapy.

Methotrexate (MTX) as an anti-inflammatory drug for the treatment of rheumatoid arthritis (RA) through oral and injectable administration is still problematic in the clinic. Herein, a MTX-loaded thermal-responsible flexible liposome (MTFL) incorporated within a carbomer-based gel was prepared as a novel transdermal agent (MTFL/Gel) for effective treatment of RA. It was found that MTFL had an average size of approximately 90 nm, which could rapidly release the drug under thermal conditions. The prepared MTFL/Gel could remarkably increase the MTX skin permeation as compared with free MTX, which was possibly due to the deformable membrane of flexible liposomes. Moreover, the results suggested MTFL/Gel could lead to a remarkably enhanced RA treatment when in combination with microwave hyperthermia. The superior ability of MTFL/Gel to alleviate RA response was attributed to the excellent skin permeation, thermal-responsible drug release, and synergistic anti-arthritic effect of MTX chemotherapy and microwave-induced hyperthermia therapy. Overall, the MTFL/Gel with dual deformable and thermal-responsible performances could be used as a novel promising transdermal agent for enhanced treatment of RA.

Design, evaluation and in vivo pharmacokinetic study of a cationic flexible liposomes for enhanced transdermal delivery of pramipexole

Pramipexole is a widely preferred antiparkinsonian agent because of its several advantages, one being the delayed onset of motor fluctuations and dyskinesia observed with other antiparkinsonian drugs. However, it may cause pulsatile stimulation of the dopamine receptors responsible for the majority of the side effects such as orthostatic hypotension, constipation, dizziness, drowsiness, dry mouth, etc. Thus, the objective of the present work was to develop flexible liposomes of pramipexole for enhanced transdermal drug delivery and consequently to reduce side effects. Structure of phospholipids, edge activator, and surfactants being major determinants of liposomes, phospholipids of varying hydrocarbon chain length and degree of substitution, and cationic and anionic surfactants (sodium hexadecyl sulfate or cetylpyridinium chloride) were used for formulating flexible liposomes. All the batches of flexible liposomes of pramipexole were prepared by the thin-film hydration method. Liposomes were characterized by TEM, % drug entrapment, vesicle size, and deformability index. Liposomes were incorporated into the gel base and formulated as a transdermal patch. Ex-vivo drug release studies demonstrated greater and faster drug releases from all the prepared cationic flexible liposomal (CFL) gel patches as compared to flexible and anionic flexible liposomal gel patches. Further, the flexible liposomes of shorter carbon chain length and monosubstitution provide rapid drug release, while the sustained release was observed through liposomes comprising of longer carbon chain length and increased substitution. Pharmacokinetic studies demonstrated that the designed transdermal patches were capable of delivering pramipexole at the desired concentration and overcome the skin barrier function efficiently. Hence, CFL transdermal patch prepared with phospholipids containing higher carbon content appears to be promising to deliver pramipexole by the transdermal route and can thus overcome the pulsatile stimulation of dopamine receptors and associated side effects.