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 Liposomal Drug Delivery System

Abstract: A liposome is the drug delivery system used for the administration
of

various types of drugs, or active substance is essential for the treatment of

various

types of disease. A liposome is a very effective drug delivery system to Target

the

active medicament to an effective part of the body without entrapping or

affecting

the other body part; that’s why it is also called the targeted drug delivery system.

Liposomes are available in various sizes to the range for treatment to various
types

of disease as the carrier for targeted the medicament or drug to active site at a

predetermined rate and time range, without affecting the other body part for the

treatment of a particular disease. They are colloidal spheres of cholesterol non-

poisonous surfactants, sphingolipids, glycolipids, long-chain unsaturated fats,

and

even layer proteins and active atoms or It is also called vesicular system. This

review discusses the advantages and disadvantages, various methods of

preparation, evaluation, etc.

Introduction: The name liposome is derived from two Greek words: Lipo =

“fat” and Soma = “body”. A liposome is the drug delivery system which is

structurally seeing like a colloidal, vesicular and made up one or more than one

lipid bilayer (outer layer) in which the equal number of aqueous layer (inner
layer)

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is inclosed into it which contains a substance like peptides and protein,
hormones,

enzymes, antibiotics, antifungal and anticancer agent .in this delivery system
drug

achieve the long therapeutic effect for the treatment of particular disease without

affected to another part of the body.

Advantages of liposomes: Liposomes are biocompatible, completely

biodegradable, non-toxic, flexible, and nonimmunogenic for systemic and

nonsystemic administrations.

• Provide controlled and sustained release.

• It carries both water and lipid-soluble drugs.

• The drug can be stabilized from oxidation.

• Targeted drug delivery or site-specific drug delivery.

• Control hydration.

Disadvantages of liposome:

• Less stability.

• Low solubility.

• Short half-life.

• High production cost.

• Leakage and fusion of encapsulated drug/mole.

Types of Liposomes:

Liposomes are classified based on –

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Based on Structural Parameters:

Unilamellar vesicles

• Small unilamellar vesicles (SUV)

• Medium unilamellar vesicles (MUV)

• Large unilamellar vesicles (LUV)

Oligolamellar vesicles (OLV)

These are made up of 2-10 bilayers of lipids surrounding a large internal volume.

Multilamellar vesicles (MLV)

MLV made up of several bilayers. And the preparation method differs from other vesicle

Preparation methods. They are arranged in a manner like an onion in which concentric
spherical

Bilayers of LUV/MLV enclosing a large number of SUV etc.

Based on Method of Preparation:

1. Reverse phase evaporation method (REV): single or oligolamellar vesicles.

2. MLV-REV: multilamellar vesicles made by a reverse-phase evaporation method.

3. SPLV: stable multilamellar vesicles

4. FATMLV: frozen and thawed MLV

5. VET: vesicles prepared by the extrusion method.

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Based upon Composition and Application:

1. Conventiuonal liposome.

2. Fusogenic liposomes (RSVE): reconstituted Sendai virus envelopes.

3. pH-sensitive liposomes.

4. Cationic liposomes.

5. Long circulatory liposomes (LCL)

6. Immune-liposomes.

Material And Method:

Composition of liposomes: The major structural components are used in the preparation

of Liposomes are given below.

1. Phospholipids

Phospholipids are the main component for the preparation of the liposome membrane.
And

They are further categorized into two category –

• Natural Phospholipids:-

◊ egg phosphatidylcholine.

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 ◊ Soybean phosphatidylcholine.

• Synthetic Phospholipids:-

◊ Dipalmitoyl phosphatidyl cloline (DPPC).

◊ Dipalmitoyl phosphatidyl ethanolamine (DPPE)

◊ Distearoyl phosphatidylserine (DSPC).

◊ Dipalmitoyl phosphatidic acid (DPPA).

◊ Dioleoyl phosphatidylglycerol (DOPG).

2. Sterols (Cholesterol)

Cholesterol and its derivatives are often used in the preparation of liposomes, and its help

too –

• reducing the permeability of the membrane to a water-soluble molecule.

• Decreasing the fluidity or microviscosity of the bilayer

• Stabilizing the membrane in the presence of biological fluids such as plasma (this effect

is used in the formulation of I .V. Liposomes).

3. Sphingolipids

Sphingosine is one of the most important parts of sphingolipids. Sphingolipids are

obtained

From plant and animal cells. Eg:- Sphingomyelin and glycosphingolipids.

4. Cationic lipids

Eg:- DODAB/C (dioctadecyl dimethyl ammonium bromide or Chloride, DOTAP

Dioleoyl

Propyl trimethyl ammonium chloride.

5. Other substances

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 • A variety of other lipids of surfactants are used to form liposomes.

• Many single-chain surfactants are used to form liposomes by mixing with cholesterol.

• Non-ionic lipids

6. Polymeric material.

Classification of liposomes

Liposomes are classified into different types-

1. According to their size.

2. According to their number of lamellae (lipid bilayer).

Method of preparation of liposomes

Two methods are used for the preparation of liposomes are:-

1. A general method of preparation

2. A specific method of preparation:- are two types-

a. Passive loading technique:

1. Mechanical dispersion

• Lipid hydration method

• Micro emulsification

• Sonication

• French pressure cell method

• Membrane extrusion

• Dried reconstituted vesicles

• Freeze-thaw method

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 2. Solvent dispersion

• Ethanol injection method

• Ether infusion method

• Double emulsification

• Reverse-phase evaporation

3. Detergent removal

b. Active loading technique

1. Prollposome

2. Lyophilization.

A general method of preparation:- In all the methods which are

used for the preparation of liposomes are involved basic four stages

are:-

• Drying down lipids from an organic solvent.

• Dispersing the lipid in aqueous media.

• Purifying the final product.

• Analyzing the final product.

a. Passive loading technique

1. Mechanical dispersion

• Lipid hydration method:-

◊ on this method firstly prepare the homogeneous mixture of

lipids. By dissolving and mixing a lipid component in an organic

solvent (chloroform)

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 ◊ Ones the lipid is thoroughly mixed in the organic solvent, the

solvent is removed to yield a lipid film.

◊ The lipid film is thoroughly dried by placing the vial or flask on

a vacuum pump overnight by removing the residual organic

solvent.

◊ Lipid solution was frozen by placing the container on a block of

dry ice or swirling the container in dry ice- acetone or alcohol.

• Micro emulsification:-

In this method, small vesicles are prepared by micro emulsifying

lipid composition using high shearing stress generated from high-

pressure homogenizer. (speed of rotation 20 to 200 for biological).

These methods are used to prepare the small lipids vesicles on a

commercial scale.

• Sonication:-

In this method lipids (MLVs) are sonicated with the help of a

sonicator in this method two types of sonicators are used either

bath type sonicator or probe sonicator for the preparation of

Liposome vesicles.

• French pressure cell method:-

This technique is basic, quick, reproducible; furthermore, it

includes delicate treatment of temperamental materials. In this

method, MLV expulsion through a small hole point at 20,000psi at

temp 4°C. Furthermore, it likewise has a few focal points over the

sonication technique. What’s more, a few disservices resemble hard

to accomplishing temperature and less working volume.

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 • Membrane extrusion:-

In this method the processed liposome has a narrow size distribution

and selected average size less than about 0.4 microns.

• Dried reconstituted vesicles

In this method, liposomes are added to an aqueous solution

containing a drug or mixed with a lyophilized protein, followed by

dehydration or mixture.

• Freeze-thaw Method.

In this method the SUVs are quickly solidified, followed by moderate

defrosting.

2. Solvent dispersion

• Ethanol injection method:-

In this method MLVs are formed by a lipid solution of ethanol are

rapidly injected into an excess of a buffer. But some drawbacks

of this method are the particles may be with heterogeneous size

distribution (30-110).

• Ether infusion method

In this method, liposomes are prepared by dissolving a lipid solution

in diethyl or ether methanol, and then the mixture is slowly injected

into an aqueous solution of a drug, to be encapsulated at a temp

of 55-65°C under the reduced pressure.

• Double emulsification

This method firstly prepared the emulsion by dissolving the drug

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 in the aqueous phase(W1), which is then emulsified in an organic

solvent of a polymer is called primary emulsion(W/O). After that

this primary emulsion further mixed in an emulsifier-containing

aqueous solution(W2) to make a W1/O/W2 double emulsion and

after than microspheres are obtained by removal of the solvent

and filtration process.

• Reverse-phase evaporation

The lipid blend is taken in a round base flagon followed by the

expulsion of dissolvable under diminished pressure by a rotational

evaporator. The framework is cleaned with nitrogen and the lipid are

re-broken down in the natural stages. The opposite stages vesicles will

frame in these stages. The standard solvent utilized is diethyl ether

and isopropyl ether. The fluid stage which contains medication to be

epitomized is included after the lipids are re-scattered in these stages.

The framework is held under persistent nitrogen; what’s more, the two

stages framework is sonicated until the blend turns out to be clear

one-stage scattering. The blend is at that point set on the rotating

evaporator, and the expulsion of natural dissolvable is done until a

gel is shaped trailed by evacuation of non-embodied material. The

subsequent liposomes are called switch stage dissipation vesicles.

3. Detergent removal

Detergents are used to solubilize the lipids at their critical micellar

concentrations. LUVs are shaped by eliminating the detergent by

dialysis and combining the micelles. In this method, the liposomes

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 are formed in homogenous size and the retention of detergent

contaminants is the drawback of this method.

b. Active loading technique

1. Prollposome:- lipid and active substances (drug) are covered

onto a solvent transporter to shape free-streaming granular

material in supportive of liposomes which structure an

isotonic liposomal suspension on hydration. The favorable to

pro- liposome approach may give a chance for cost-effective

large scale manufacturing liposomes containing particularly

lipophilic drugs.

2. Lyophilization:-the expulsion of water from items in a solidified

state at incredibly decreased weight is called lyophilization

(freeze-drying). The cycle is commonly used to dry items that

are thermolabile which might be annihilated by heat-drying.

This method has an incredible potential to unravel long

haul steadiness issues as for liposomal solidness. Spillage of

entangled materials may occur during the cycle of freeze-drying

and on reconstitution.

Result and Discussion:

Evaluation of liposomes

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 The purpose of the evaluation of liposome to ensure the in vivo

and in vitro performance of liposomes. Evaluation parameter are

categories into three broad categories are-

• Physical characterization:- Size, shape, surface feature,

lamellarty, phase behavior and drug release profile.

• Chemical characterization

• Biological characterization:- To ensure the safety and suitability

of formulation for therapeutic use.

Some parameters are

Vesicle shape and lamellarity

Vesicle shape can be evaluated utilized electron microscopic

techniques. Lamellarity of vesicles for example number of bilayers

present in liposomes is decided to utilize freeze-fracture electron

microscopy and p-31 nuclear magnetic resonance analysis.

Vesicle size and size distribution

Various techniques are used to describe the size and size distribution

for eg. Light microscopy, fluorescent microscopy, electron

microscopy, laser light scattering photon correlation spectroscopy,

field flow fractionation, gel permeation, and exclusion. The

most exact strategy for deciding the size of liposome is electron

microscopy since it licenses one to see ever individual liposomes

and to acquire accurate data about the profile of liposome. Most

of the evaluation methods are used for liposome are categorize

into a various category:-

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 Microscopic, diffraction, scattering, and hydrodynamic techniques.

Encapsulation efficiency and trapped volume

This determines the amount and rate of entrapment of water-

soluble agents in the aqueous compartment of the liposome.

Drug release

The component of medication discharge from liposome can be

surveyed by utilizing very much aligned in vitro dispersion cells.

The liposome-based definition can be helped by utilizing in vitro

tests to foresee pharmacokinetics and bioavailability of medication

previously utilizing expensive and tedious in vivo examinations.

The weakening actuated medication discharge in cushion and

plasma was utilized as an indicator for pharmacokinetic execution of

liposomal details and another measure which decided intracellular

medication discharge prompted by liposome debasement in the

presence of mouse-liver lysosome lysate was utilized to evaluate

the bioavailability of medication.

Application of liposome:

The application of liposomes are categorized into two categories are:

• general application:- in this category, liposomes are used to

treat respiratory disorders, eye disorders, vaccine adjuvant,

brain targeting, and anti-infective agents.

• clinical application:-

◊ cancer chemotherapy.

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 ◊ gene therapy.

◊ liposomes for topical application.

◊ liposomes as a carrier of the drug in oral treatment.

◊ Liposomes for pulmonary delivery.

◊ Against leishmaniasis.

◊ Cell biological application.

◊ Metal storage disease.

◊ Ophthalmic delivery of drugs.

Conclusion:

Liposome has been acknowledged as an incredibly valuable

transporter framework for focused medication conveyance. The

adaptability of their conduct can be used for the medication

conveyance through any course of the organization and for any

medication material regardless.

Refernces:

1. Mazur, F.; Bally, M.; Städler, B.; Chandrawati, R. Liposomes and lipid
bilayers

in biosensors. Adv. Colloid Interface Sci. 2017, 249, 88–99.

2. Düzgüne¸s, N.; Gregoriadis, G. Introduction: The Origins of Liposomes: Alec

Bangham at Babraham. In Methods in Enzymology; Academic Press: Cambridge,

16
 MA, USA, 2005; Volume 391, pp. 1–3.

3. Bangham, A.D.; Horne, R.W. Negative staining of phospholipids and their

structural modification by surface-active agents as observed in the electron

microscope. J. Mol. Biol. 1964, 8, 660–668.

4. Mirzavi, F.; Barati, M.; Soleimani, A.; Vakili-Ghartavol, R.; Jaafari, M.R.;

Soukhtanloo, M. A review on liposome-based therapeutic Approaches against

malignant melanoma. Int. J. Pharm. 2021, 599, 120413.

5. Wang, G.; Li, R.; Parseh, B.; Du, G. Prospects and challenges of anticancer

agents’ delivery via chitosan-based drug carriers to Combat breast cancer: A

review. Carbohydr. Polym. 2021, 268, 118192.

6. Watson, D.S.; Endsley, A.N.; Huang, L. Design considerations for liposomal

vaccines: Influence of formulation parameters on antibody and cell-mediated

immune responses to liposome associated antigens. Vaccine 2012, 30, 2256–2272.

7. Man, F.; Gawne, P.J.; de Rosales, R.T.M. Nuclear imaging of liposomal drug

delivery systems: A critical review of radiolabelling methods and applications in

nanomedicine. Adv. Drug Delivery Rev. 2019, 143, 134–160.

8. Dos Santos Rodrigues, B.; Banerjee, A.; Kanekiyo, T.; Singh, J.

Functionalized

liposomal nanoparticles for efficient gene delivery system to neuronal cell

transfection. Int. J. Pharm. 2019, 566, 717–730.

9. Taha, E.I.; El-Anazi, M.H.; El-Bagory, I.M.; Bayomi, M.A. Design of

liposomal colloidal systems for ocular delivery of ciprofloxacin. Saudi Pharm. J.

2014, 22, 231–239.

10. Han, Y.; Gao, Z.; Chen, L.; Kang, L.; Huang, W.; Jin, M.; Wang, Q.; Bae,

Y.H. Multifunctional oral delivery systems for enhanced bioavailability of

17
 therapeutic peptides/proteins. Acta Pharm. Sin. B 2019, 9, 902–922.

11. Mirtaleb, M.S.; Shahraky, M.K.; Ekrami, E.; Mirtaleb, A. Advances in

biological nano-phospholipid vesicles for transdermal delivery: A review on

applications. J. Drug Delivery Sci. Technol. 2021, 61, 102331.

12. Mehta, P.P.; Ghoshal, D.; Pawar, A.P.; Kadam, S.S.; Dhapte-Pawar, V.S.

Recent advances in inhalable liposomes for treatment of Pulmonary diseases:

Concept to clinical stance. J. Drug Delivery Sci. Technol. 2020, 56, 101509.

13. Yusuf, H.; Ali, A.A.; Orr, N.; Tunney, M.M.; Mc Carthy, H.O.; Kett, V.L.

Novel freeze-dried DDA and TPGS liposomes are suitable for nasal delivery of

vaccine. Int. J. Pharm. 2017, 533, 179–186.

14. Liu, W.; Hou, Y.; Jin, Y.; Wang, Y.; Xu, X.; Han, J. Research progress on

liposomes: Application in food, digestion behavior and absorption mechanism.

Trends Food Sci. Technol. 2020, 104, 177–189.

15. Himeno, T.; Konno, Y.; Naito, N. Liposomes for Cosmetics. In Cosmetic

Science and Technology; Sakamoto, K., Lochhead, R.Y.,Maibach, H.I.,

Yamashita,

Y., Eds.; Elsevier: Amsterdam, The Netherlands, 2017; pp. 539–549.

16. Niu, M.; Lu, Y.; Hovgaard, L.; Guan, P.; Tan, Y.; Lian, R.; Qi, J.; Wu, W.

Hypoglycemic activity and oral bioavailability of Insulin-loaded liposomes

containing bile salts in rats: The effect of cholate type, particle size and

administered dose. Eur. J. Pharm.Biopharm. 2012, 81, 265–272.

17. Wang, N.; Wang, T.; Li, T.; Deng, Y. Modulation of the physicochemical
state

of interior agents to prepare controlled release Liposomes. Colloids Surf. B 2009,

69, 232–238.

18
 18. Zeng, H.; Qi, Y.; Zhang, Z.; Liu, C.; Peng, W.; Zhang, Y. Nanomaterials

toward the treatment of Alzheimer’s disease: Recent Advances and future trends.

Chin. Chem. Lett. 2021, 32, 1857–1868.

19. Li, C.; Zhang, Y.; Wan, Y.; Wang, J.; Lin, J.; Li, Z.; Huang, P. STING-

activating drug delivery systems: Design strategies and Biomedical applications.

Chin. Chem. Lett. 2021, 32, 1615–1625.

20. Forssen, E.A. The design and development of DaunoXome® for solid tumor

targeting in vivo. Adv. Drug Delivery Rev. 1997, 24,133–150.

21. Kalyane, D.; Raval, N.; Maheshwari, R.; Tambe, V.; Kalia, K.; Tekade, R.K.

Employment of enhanced permeability and retention Effect (EPR): Nanoparticle-

based precision tools for targeting of therapeutic and diagnostic agent in cancer.

Mater. Sci. Eng. C Mater. Biol Appl. 2019, 98, 1252–1276.

22. Zhang, M.; Gao, S.; Yang, D.; Fang, Y.; Lin, X.; Jin, X.; Liu, Y.; Liu, X.; Su,

K.; Shi, K. Influencing factors and strategies of enhancing Nanoparticles into

tumors in vivo. Acta Pharm. Sin. B. 2021, 11, 2265–2285.

23. Dana, P.; Bunthot, S.; Suktham, K.; Surassmo, S.; Yata, T.; Namdee, K.;

Yingmema, W.; Yimsoo, T.; Ruktanonchai, U.R.; Sathornsumetee, S.; et al.

Active

targeting liposome-PLGA composite for cisplatin delivery against cervical cancer.

Colloids Surf. B Biointerfaces 2020, 196, 111270.

24. Hashemi, M.; Shamshiri, A.; Saeedi, M.; Tayebi, L.; Yazdian-Robati, R.

Aptamer-conjugated PLGA nanoparticles for delivery and Imaging of cancer

therapeutic drugs. Arch. Biochem. Biophys. 2020, 691, 108485.

25. Fernandes, M.A.; Eloy, J.O.; Luiz, M.T.; Junior, S.L.R.; Borges, J.C.; de la

Fuente, L.R.; Luis, C.O.S.; Marchetti, J.M.; Santos-Martinez, M.J.; Chorilli, M.

19
 Transferrin-functionalized liposomes for docetaxel delivery to prostate cancer
cells.

Colloids Surf. A 2021, 611, 125806.

26. Danhier, F.; Breton, A.L.; Preat, V. RGD-based strategies to target alphav

beta3 integrin in cancer therapy and diagnosis. Mol. Pharm. 2012, 9, 2961–2973.

27. Kang, T.; Gao, X.; Hu, Q.; Jiang, D.; Feng, X.; Zhang, X.; Song, Q.; Yao, L.;

Huang, M.; Jiang, X.; et al. iNGR-modified PEG-PLGA Nanoparticles that

recognize tumor vasculature and penetrate gliomas. Biomaterials 2014, 35, 4319–

4332.

28. Liang, H.; Zou, F.; Liu, Q.; Wang, B.; Fu, L.; Liang, X.; Liu, J.; Liu, Q.

Nanocrystal-loaded liposome for targeted delivery of poorly water-soluble

antitumor drugs with high drug loading and stability towards efficient cancer

therapy. Int. J. Pharm. 2021, 599, 120418. [CrossRef]

29. Chen, Q.; Gao, M.; Li, Z.; Xiao, Y.; Bai, X.; Boakye-Yiadom, K.O.; Xu, X.;

Zhang, X.-Q. Biodegradable nanoparticles decorated with different carbohydrates

for efficient macrophage-targeted gene therapy. J. Control. Release 2020, 323,

179–

190.

30. Pattni, B.S.; Chupin, V.V.; Torchilin, V.P. New Developments in Liposomal

Drug Delivery. Chem. Rev. 2015, 115, 10938–10966.

31. Kim, T.; Kim, J.; Kim, S. Extended-release formulation of morphine for

subcutaneous administration. Cancer Chemother. Pharmacol. 1993, 33, 187–190.

32. Fan, Y.; Marioli, M.; Zhang, K. Analytical characterization of liposomes and

other lipid nanoparticles for drug delivery. J. Pharm.Biomed. Anal. 2021, 192,

20
 113642.

33. Wang, N.; Chen, M.; Wang, T. Liposomes used as a vaccine adjuvant-delivery

system: From basics to clinical immunization. J. control. Release 2019, 303, 130–

150.

34. Barenholz, Y. Doxil®—The first FDA-approved nano-drug: Lessons learned.

J. Control. Release 2012, 160, 117–134.

35. Dicko, A.; Kwak, S.; Frazier, A.A.; Mayer, L.D.; Liboiron, B.D. Biophysical

characterization of a liposomal formulation of cytarabine and daunorubicin. Int. J.

Pharm. 2010, 391, 248–259.

36. Ye, Q.; Asherman, J.; Stevenson, M.; Brownson, E.; Katre, N.V. DepoFoam™

technology: A vehicle for controlled delivery of protein and peptide drugs. J.

Control. Release 2000, 64, 155–166.

37. Large, D.E.; Abdelmessih, R.G.; Fink, E.; Auguste, D.T. Liposome
composition

in drug delivery design, synthesis, characterization, and clinical application. Adv.

Drug Delivery Rev. 2021, 176, 113851.

38. Guimarães, D.; Cavaco-Paulo, A.; Nogueira, E. Design of liposomes as drug

delivery system for therapeutic applications. Int. J. Pharm. 2021, 601, 120571.

39. He, Y.; Qin, L.; Huang, Y.; Ma, C. Advances of Nano-Structured Extended-

Release Local Anesthetics. Nanoscale Res. Lett. 2020,15, 13.

40. Hillery, A.M. Supramolecular lipidic drug delivery systems: From laboratory
to

clinic A review of the recently introduced commercial liposomal and lipid-based

formulations of amphotericin B. Adv. Drug Delivery Rev. 1997, 24, 345–363.

41. Beiranvand, S.; Eatemadi, A.; Karimi, A. New Updates Pertaining to Drug

21
 Delivery of Local Anesthetics in Particular Bupivacaine using Lipid
Nanoparticles.

Nanoscale Res. Lett. 2016, 11, 307–317.

42. Richter, A.M.; Waterfield, E.; Jain, A.K.; Canaan, A.J.; Allison, B.A.; Levy,

J.G. Liposomal delivery of a photosensitizer, benzopor-phyrin derivative

monoacid

ring A (BPD), to tumor tissue in a mouse tumor model. Photochem. Photobiol.

1993, 57, 1000–1006.

43. Alving, C.R.; Beck, Z.; Matyas, G.R.; Rao, M. Liposomal adjuvants for
human

vaccines. Expert Opin. Drug Deliv. 2016, 13, 807–816.

44. Signorell, R.D.; Luciani, P.; Brambilla, D.; Leroux, J.C. Pharmacokinetics of

lipid-drug conjugates loaded into liposomes. Eur. J. Pharm. Biopharm. 2018, 128,

188–199.

45. Nogueira, E.; Gomes, A.C.; Preto, A.; Cavaco-Paulo, A. Design of liposomal

formulations for cell targeting. Colloids Surf. B. 2015,136, 514–526.

46. Kohli, A.G.; Kierstead, P.H.; Venditto, V.J.; Walsh, C.L.; Szoka, F.C.
Designer

lipids for drug delivery: From heads to tails. J.control. Release 2014, 190, 274–
287.

47. Liu, Y.; Mei, Z.; Mei, L.; Tang, J.; Yuan, W.; Srinivasan, S.; Ackermann, R.;

Schwendeman, A.S. Analytical method development and comparability study for

AmBisome® and generic Amphotericin B liposomal products. Eur. J. Pharm.

Biopharm. 2020, 157, 241–249.

48. Takechi-Haraya, Y.; Matsuoka, M.; Imai, H.; Izutsu, K.; Sakai-Kato, K.

Detection of material-derived differences in the stiffness of egg yolk

22
 phosphatidylcholine-containing liposomes using atomic force microscopy. Chem.

Phys. Lipids 2020, 233, 104992.

49. Li, J.; Wang, X.; Zhang, T.; Wang, C.; Huang, Z.; Luo, X.; Deng, Y. A review

on phospholipids and their main applications in drug delivery systems. Asian J.

Pharm. Sci. 2015, 10, 81–98.

50. Luo, R.; Li, Y.; He, M.; Zhang, H.; Yuan, H.; Johnson, M.; Palmisano, M.;

Zhou, S.; Sun, D. Distinct biodistribution of doxorubicinb and the altered

dispositions mediated by different liposomal formulations. Int. J. Pharm. 2017,

519,

1–10.

51. Skupin-Mrugalska, P.; Piskorz, J.; Goslinski, T.; Mielcarek, J.; Konopka, K.;

Düzgüne¸s, N. Current status of liposomal porphyrinoid photosensitizers. Drug

Discov. Today 2013, 18, 776–784.

52. Saraf, S.; Jain, A.; Tiwari, A.; Verma, A.; Panda, P.K.; Jain, S.K. Advances in

liposomal drug delivery to cancer: An overview. J. Drug Deliv. Sci. Technol.

2020,

56, 101549.

53. Garbuzenko, O.; Barenholz, Y.; Priev, A. Effect of grafted PEG on liposome

size and on compressibility and packing of lipid bilayer. Chem. Phys. Lipids 2005,

135, 117–129.

54. Song, L.Y.; Ahkong, Q.F.; Rong, Q.; Wang, Z.; Ansell, S.; Hope, M.J.; Mui,
B.

Characterization of the inhibitory effect of PEG-lipid conjugates on the

intracellular

delivery of plasmid and antisense DNA mediated by cationic lipid liposomes.

23
 Biochim. Biophys. Acta Biomembr. 2002, 1558, 1–13.

55. Varga, Z.; Wacha, A.; Vainio, U.; Gummel, J.; Bóta, A. Characterization of
the

PEG layer of sterically stabilized liposomes: A SAXS study. Chem. Phys. Lipids

2012, 165, 387–392.

56. Kim, S.; Howell, S.B. Multivesicular Liposomes Having a Biologically Active

Substance Encapsulated Therein in the Presence of a hydrochloride. U.S. Patent

5,723,147, 3 March 1998.

57. Mantripragada, S. A lipid based depot (DepoFoam® technology) for sustained

release drug delivery. Prog. Lipid Res. 2002, 41,392–406.

58. Perkins, W.; Malinin, V.; Li, X.; Miller, B.; Seidel, D.; Holzmann, P.;

Schulz, H.; Hahn, M. System for Treating Pulmonary Infections.U.S. Patent

9,566,234 B2, 14 February 2017.

59. Borochov, H.; Shinitzky, M.; Barenholz, Y. Sphingomyelin phase transition

in the sheep erythrocyte membrane. Cell Biochem. Biophys. 1979, 1, 219–228.

60. Vemuri, S.; Rhodes, C.T. Preparation and characterization of liposomes as

therapeutic delivery systems: A review. Pharm. Acta Helv. 1995, 70, 95–111.

61. Takechi-Haraya, Y.; Sakai-Kato, K.; Abe, Y.; Kawanishi, T.; Okuda, H.;
Goda,

Y. Atomic Force Microscopic Analysis of the Effect of Lipid Composition on

Liposome Membrane Rigidity. Langmuir 2016, 32, 6074–6082.

62. Pajewski, R.; Djedoviˇc, N.; Harder, E.; Ferdani, R.; Schlesinger, P.H.; Gokel,

G.W. Pore formation in and enlargement of phospholipid liposomes by synthetic

models of ceramides and sphingomyelin. Bioorg. Med. Chem. 2005, 13, 29–37.

63. Webb, M.S.; Bally, M.B.; Mayer, L.D.; Miller, J.J.; Tardi, P.G.
Sphingosomes

24
 for Enhanced Drug Delivery. U.S. Patent 5,741,516, 21 April 1998.

64. Silverman, J.A.; Deitcher, S.R. Marqibo® (vincristine sulfate liposome

injection) improves the pharmacokinetics and pharmacody-namics of vincristine.

Cancer Chemother. Pharmacol. 2013, 71, 555–564.

65. Kaddah, S.; Khreich, N.; Kaddah, F.; Charcosset, C.; Greige-Gerges, H.

Cholesterol modulates the liposome membrane fluidity and permeability for a

hydrophilic molecule. Food Chem. Toxicol. 2018, 113, 40–48.

66. Bhattarai, A.; Likos, E.M.; Weyman, C.M.; Shukla, G.C. Regulation of

cholesterol biosynthesis and lipid metabolism: A microRNA management

perspective. Steroids 2021, 173, 108878.

67. Sadeghi, N.; Deckers, R.; Ozbakir, B.; Akthar, S.; Kok, R.J.; Lammers, T.;

Storm, G. Influence of cholesterol inclusion on the doxorubicin release

characteristics of lysolipid-based thermosensitive liposomes. Int. J. Pharm. 2018,

548, 778–782.

68. Wang, M.; Liu, M.; Xie, T.; Zhang, B.; Gao, X. Chitosan-modified
cholesterol-

free liposomes for improving the oral bioavailability of progesterone. Colloids

Surf.

B. 2017, 159, 580–585.

69. Kirby, C.; Gregoriadis, G. The effect of the cholesterol content of small

unilamellar liposomes on the fate of their lipid components in vivo. Life Sci.
1980,

27, 2223–2230.

70. Najafinobar, N.; Mellander, L.J.; Kurczy, M.E.; Dunevall, J.; Angerer, T.B.;

Fletcher, J.S.; Cans, A. Cholesterol Alters the Dynamics of Release in Protein

25
 Independent Cell Models for Exocytosis. Sci. Rep. 2016, 6, 33702–33712.

71. Garcon, N.M.C.; Friede, M. Vaccines Contraining a Saponin and a Sterol.

U.S.

Patent US2005/0214322A1, 29 September 2005.

72. Abboud, R.; Greige-Gerges, H.; Charcosset, C. Effect of Progesterone, Its

Hydroxylated and Methylated Derivatives, and Dydrogesterone on Lipid Bilayer

Membranes. J. Membrane Biol. 2015, 248, 811–824.

73. Kapoor, M.; Lee, S.L.; Tyner, K.M. Liposomal Drug Product Development
and

Quality: Current US Experience and Perspective. AAPS J. 2017, 19, 632–641.

74. Adler-Moore, J.; Gamble, R.C.; Proffitt, R.T. Treatment of Systemic Fungal

Infections with Phospholipid Particles Encapsulating Polyene Antibiotics. U.S.

Patent 5,874,104, 23 February 1999.

75. Lu, B.; Ma, Q.; Zhang, J.; Liu, R.; Yue, Z.; Xu, C.; Li, Z.; Lin, H. Preparation

and characterization of bupivacaine multivesicular Liposome: A QbD study about

the effects of formulation and process on critical quality attributes. Int. J. Pharm.

2021, 598, 120335.

76. Sala, M.; Miladi, K.; Agusti, G.; Elaissari, A.; Fessi, H. Preparation of

liposomes: A comparative study between the double solvent displacement and the

conventional ethanol injection—From laboratory scale to large scale. Colloids

Surf.

A 2017, 524, 71–78.

77. Boni, L.T.; Miller, B.S.; Malinin, V.; Li, X. Sustained Release of
Antinfectives.

U.S. Patent 8,802,137B2, 12 August 2014.

26
 78. Catherine, C.; Audrey, J.; Jean-Pierre, V.; Sebastien, U.; Hatem, F.
Preparation

of liposomes at large scale using the ethanol injection Method: Effect of scale-up

and injection devices. Chem. Eng. Res. Des. 2015, 94, 508–515.

79. Laouini, A.; Jaafar-Maalej, C.; Sfar, S.; Charcosset, C.; Fessi, H. Liposome

preparation using a hollow fiber membrane contactor— application to

spironolactone encapsulation. Int. J. Pharm. 2011, 415, 53–61.

80. Wagner, A.; Vorauer-Uhl, K.; Kreismayr, G.; Katinger, H. The crossflow

injection technique: An improvement of the ethanol injection method. J. Liposome

Res. 2002, 12, 259–270.

81. Wagner, A.; Vorauer-Uhl, K.; Katinger, H. Liposomes produced in a pilot

scale: Production, purification and efficiency aspects. Eur. J. Pharm. Biopharm.

2002, 54, 213–219.

82. Gouda, A.; Sakr, O.S.; Nasr, M.; Sammour, O. Ethanol injection technique

for liposomes formulation: An insight into development,influencing factors,

challenges and applications. J. Drug Delivery Sci. Technol. 2021, 61, 102174.

83. Schubert, M.A.; Müller-Goymann, C.C. Solvent injection as a new approach

for manufacturing lipid nanoparticles – evaluation of the method and process

parameters. Eur. J. Pharm. Biopharm. 2003, 55, 125–131.

84. Utsugil, T.; Nii, A.; Fan, D.; Pak, C.C.; Denkins, Y.; Hoogevest, P.V.; Fidler,

I.J. Comparative efficacy of liposomes containing synthetic bacterial cell wall

analogues for tumoricidal activation of monocytes and macrophages. Cancer

Immunol. Immunother. 1991, 33, 285–292.

85. Frost, H. MTP-PE in liposomes as a biological response modifier in the

treatment of cancer: Current status. Biotherapy 1992, 4, 199–204.

27
 86. Sone, S.; Utsugi, T.; Tandon, P.; Ogawara, M. A dried preparation of
liposomes

containing muramyl tripeptide phos-phatidylethanolamine as a potent activator of

human blood monocytes to the antitumor state. Cancer Immunol. Immunother.

1986, 22, 191–196.

87. Barenholzt, Y.; Amselem, S.D.L. A new method for preparation of

phospholipid vesicles (liposomes)—french press. FEBS Lett. 1979, 99, 210–214.

88. Castile, J.D.; Taylor, K.M.G. Factors affecting the size distribution of

liposomes produced by freeze–thaw extrusion. Int. J. Pharm. 1999, 188, 87–95.

89. Pupo, E.; Padrón, A.; Santana, E.; Sotolongo, J.; Quintana, D.; Dueñas, S.;

Duarte, C.; de la Rosa, M.C.; Hardy, E. Preparation of plasmid DNA-containing

liposomes using a high-pressure homogenization–extrusion technique. J. Control.

Release 2005, 104, 379–396.

90. Johnson, S.M.; Bangham, A.D.; Hill, M.W.; Korn, E.D. Single bilayer

liposomes. Biochim. Biophys. Acta Biomembr. 1971, 223, 820–826.

91. Lesieur, S.; Grabielle-Madelmont, C.; Paternostre, M.T.; Ollivon, M. Size

analysis and stability study of lipid vesicles by high- performance gel exclusion

chromatography, turbidity, and dynamic light scattering. Anal. Biochem. 1991,

192,

334–343.

92. Hunter, D.G.; Frisken, B.J. Effect of Extrusion Pressure and Lipid Properties
on

the Size and Polydispersity of Lipid Vesicles. Biophys. J. 1998, 74, 2996–3002.

93. Berger, N.; Sachse, A.; Bender, J.; Schubert, R.; Brandl, M. Filter extrusion of

liposomes using different devices: Comparison of Liposome size, encapsulation

28
 efficiency, and process characteristics. Int. J. Pharm. 2001, 233, 55–68.

94. Ong, S.G.M.; Chiteni, M.; Lee, K.S.; Ming, L.C.; Yuen, K.H.Y. Evaluation of

Extrusion Technique for Nanosizing Liposomes.Pharmaceutics 2016, 8, 36.

95. Mokhtarieh, A.A.; Davarpanah, S.J.; Lee, M.K. Ethanol treatment a Non-

extrusion method for asymmetric liposome size optimization. DARU J. Pharm.

Sci.

2013, 21, 32.

96. Preksha, V.; Patel, J.K.; Patel, M.M. High-Pressure Homogenization

Techniques for Nanoparticles. In Emerging Technologies for Nanoparticle

Manufacturing; Patel, J.K., Pathak, Y.V., Eds.; Springer: Cham, Switzerland,

2021;

Part III; pp. 263–286.

97. Kyun, S.; Gye, C.; Shin, H.; Kyo, M.; In, J.; Hwang, C.; Park, J. Factors

influencing the physicochemical characteristics of cationic polymer-coated

liposomes prepared by high-pressure homogenization. Colloids Surf. A 2014, 454,

8–15.

98. Barnadas-Rodríguez, R.; Sabés, M. Factors involved in the production of

liposomes with a high-pressure homogenizer. Int. J. Pharm. 2001, 213, 175–186.

99. Proffitt, R.T.; Alder-Moore, J.; Chiang, S.M. Amphotericin B Liposome

Preparation. U.S. Patent 5,965,156, 12 October 1999.

100. Zhang, Y.; Hill, A.T. Amikacin liposome inhalation suspension as a

treatment for patients with refractory mycobacterium avium complex lung

infection. Expert Rev. Resp. Med. 2020, 15, 737–744.

101. Gadekar, V.; Borade, Y.; Kannaujia, S.; Rajpoot, K.; Anup, N.;
Tambe,

V.; Kalia, K.; Tekade, R.K. Nanomedicines accessible in the market for clinical

29
 interventions. J. Control. Release 2021, 330, 372–397.

102. Kim, S.; Kim, T.; Murdande, S. Sustained-Release Liposomal

Anesthetic Compositions. U.S. Patent 8,182,835B2, 22 May 2012.

103. Swenson, C.E.; Perkins, W.R.; Roberts, P.; Janoff, A.S. Liposome

technology and the development of Myocet™ (liposomal Doxorubicin citrate).

Breast 2001, 10, 1–7.

104. Sarris, A.H.; Cabanillas, F.; Logan, P.M.; Burge, C.T.R.; Goldie, J.H.;

Webb, M.S. Compositions and Methods for Treating Lymphoma. U.S. Patent

7,247,316 B2, 24 July 2007.

105. Mayer, L.D.; Nayar, R.; Thies, R.L.; Boman, N.L.; Cullis, P.R.; Bally,

M.B. Identification of vesicle properties that enhance the antitumour activity of

liposomal vincristine against murine L1210 leukemia. Cancer Chemoth. Pharm.

1993, 33, 17–24.

106. Nichols, J.W.; Deamer, D.W. Catecholamine uptake and concentration

by liposomes maintaining pH gradients. Biochim. Biophys. Acta 1976, 455, 269–

271.

107. Boman, N.L.; Masin, D.; Mayer, L.D.; Cullis, P.R.; Bally, M.B.

Liposomal Vincristine Which Exhibits Increased Drug Retention and Increased

Circulation Longevity Cures Mice Bearing P388 Tumors. Cancer Res. 1994, 54,

2830–2833.

108. Drummond, D.C.; Kirpotin, D.B.; Hayes, M.E.; Kesper, C.N.K.;

Awad,

A.M.; Moore, D.J.; O’Brien, A.J. Stabilizing Camptothecin Pharmaceutical

Compositions. U.S. Patent 2020179371A1, 11 June 2020.

109. Hong, K.; Drummond, D.C.; Kirpotiin, D. Liposome Useful for Drug

30
 Delivery. U.S. Patent 20160338956A1, 24 June 2016.

110. Irby, D.; Du, C.; Li, F. Lipid–Drug Conjugate for Enhancing Drug

Delivery. Mol. Pharm. 2017, 14, 1325–1338.

31