Ambikanandan Misra, Professor of Pharmaceutics & Head, Pharmacy

Department

TIFAC – CORE IN NDDSPHARMACY DEPARTMENT

FACULTY OF TECHOLOGY & ENGINEERING.KALABHAVAN

M.S. UNIVERSITY OF BARODA,VADODARA – 390 001, GUJARAT

RECENT ADVANCES IN LIPOSOMAL DRY POWDER INHALER FORMULATIONS FOR

PULMONARY DISORDER

Act quickly but short lived action

Minimize the dose required

Are non-invasive

Minimizes side effects

Avoids hepatic first-pass metabolism

Advantages of the Inhaled Dosage formAdvantages of the Inhaled Dosage form

Are systemically absorbed

Identified target

Specific drug

Excellent drug delivery system•Device •Formulation

Efficient patient and practitioner education

The Keys to Successful Pulmonary The Keys to Successful Pulmonary Drug DeliveryDrug Delivery

Ideal Therapeutic AerosolsIdeal Therapeutic Aerosols

Contain a safe and efficacious drug!

Contain minimal quantities of inert excipients

Monodisperse, small particle size

Low velocity after generation

High concentration and rate of generation

Highly reproducible characteristics

Low bioburden (solids) or sterile (liquids)

Factors Affecting Lung Deposition Factors Affecting Lung Deposition

• Inhaled ParticlesSizeShape, Density, Charge, HygroscopicityPatient

• Lung AnatomyDisease StateBreathing Pattern

• Aerosol Generation DevicePrinciple and Design Features

Deposition depicted by gamma scintigraphy from Pharmaceutical Profiles

Pulmonary Delivery of Therapeutics, Pulmonary Delivery of Therapeutics, Peptides, Proteins and Genes Peptides, Proteins and Genes

Formulation ChallengesInsoluble drugsShear sensitiveThermolabileExcipientsConformational changesInstability

Particle “Engineering”MicrospheresLiposomesPorous particlesPEGylated particles

Device PlatformsPressurized metered dose inhalers (pMDI)NebulizersDry powder inhalers (DPI)Single breath liquid systems

LUNG DISEASES WHICH CAN BE TARGETED BY LIPOSOMAL DPI

• Asthma• Chronic obstructive pulmonary disease (COPD).• Lung cancer• Cystic Fibrosis and associated lung infections• Local Pain• Pneumonia• Local fungal infection• Pulmonary Tuberculosis

DPI

CONVENTIONAL DPI NOVEL & ENGINEERED DPI

MIXING OF DRUG AND CARRIER LACTOSEAND DELIVERY TO LUNG

HAVE DISADVANTAGEOF HIGH AND REPEATED DRUG DOSING AND LOW PULMONARY BIOAVAILABILITY.

• USE OF TERNARY MIXTURES• ALTERING PHYSICAL SHAPE,

LENGTH, DENSITY OF THE CARRIER LACTOSE

• USE OF LIPOSOMES AND NANOPARTICLES AS DRUG CARRIER

• LOW DENSITY PARTICLES• USE OF LESS COHESIVE AND

ADHESIVE, WRINKLED PARTICLES.

DRY POWDER INHALERSADVANTAGES:• Automatic coordination.• No propellants.• Drug stability advantages.• High drug dose carrying capacities.• High reproducibility (Monodisperse)• Minimal extrapulmonary loss of drug due to low oropharyngeal

deposition, low device retention and low exhaled loss.

CONTEMPORARY ISSUES OF DPI:

• Effort Dependant (Potential for poor repeatability and unsuitable for young children).

• Drug deposition in deep lung for systemic absorption.Large fraction of coarse articles (Increase upper airway deposition)

• Prolonged pulmonary retention (Changed Pharmacokinetcs)

• Enhanced cellular uptake (Pharmacological response).

• Liposomal membrane composition and surface properties

• Patentability of drug.• Reduced side effects due to slow systemic

dilution.• Protection of drug from enzymatic degradation

Why Use Liposomes in Drug Delivery?

Drug Targeting:• Nanoliposomes for avoiding alveolar macrophage uptake.• Liposomes localization in Macrophages in Lung Fungal

infections.• Surface Modification through ligand (antibodies, enzymes,

protein A, sugars) in liposomes.• pH responsive liposomes for tumor specific drug release.

Why Use Liposomes in Pulmonary Drug Delivery?

Hence Lower Dose and Less Adverse Reactions.

Liposomal DPI Preparation

Lyophillization Spray drying

Use of carrier lactose, cyroprotectant, amino acids, lipids during freeze and spray drying.

Dry Liposomal Powder

Use of Ternary Mixtures and effect of fines

Altered physical shape, density,

porosity

Less cohesive and adhesive particles,

lubricants

Enhanced DPI Properties

Formulation of Liposomes

Method:•Lyophilization (Rate of cooling , drying, Vacuum )•Spray Drying•Ternary Mixtures

Characterization•Water content•Carrier ratio•Particle size•Degradation•Drug leakage•Drug release & Stability

Cryoprotectant Selection

•Type

•Ratio

Process optimization

•Freezing time

•Drying time

Process flow chart

Formulation of Dry Powder Inhalers

Liposomal Dry Powder Inhalers

In vitro characterizationEmitted dose DensityFine particle fraction (Twin Stage Impinger/ Cascade Impactor)Flowability

In vivo characterization (in animal models) Pharmacokinetic evaluation Pharmacodynamic evaluation

Process optimization

•Hydration volume•Hydration time•Sonication frequency & time

Method of Preparation•Thin film Hydration•Reverse Phase evaporation•Microfluidization and spray drying

Characterization •Drug entrapment•Entrapment Efficiency•Particle size & Drug release•Lamellarity•Stability and drug retention studies

Drug - Lipid selection

•Drug - lipid Ratio•PC-Cholesterol Ratio•Charge on liposome

Our Research Achievements and Contributions

Evaluation & Optimization of flow and dispersion (deaggregation) characteristics (drug at the desired site)

Pharmaceutical development of liposomal DPI formulations

In vivo evaluation on atleast two animal models and if possible, the clinical evaluation of the potential formulations.

Development of systemic DPI systems for polypeptides, genetically engineered drugs and other drugs of importance.

Objectives

Lung localization of drug–maximizes the therapeutic index–minimizing unwanted systemic activity or toxicity

Drug distribution limited to the intended site of action in the lung provides

- maintenance of prolonged therapeutic levels - slow systemic dilution

HypothesisPulmonary DPI

Formulations - Reduction in dose, duration of therapy and systemic side-effects- reduce the cost of therapy

Enhancement of Pulmonary absorption- Protein, peptides and genetically engineered drugs - Other drugs for compliance and ease of delivery

Antiasthamatic

Ketotifen Fumarate, Budesonide Salbutamol and Terbutaline Sulphate, MometasoneFuroate(LectinConjugated Nanoparticles)

- Spray dried Porous and Lyophillized Light and Large particles developed.

- Higher cellular uptake and enhanced antiproliferative activity in A-549 cell lines

Our Research Achievements:

Comparative pulmonary drug release from LDPI of Ketotifen

Our Research Achievements:

AntifungalAmphotericin B

- Spray dried Porous and LyophillizedLight and Large particles developed.

- In vivo Animal experimentation and clinical evaluation for set goals in progress.

Our Research Achievements continuedcontinued……2.

Proteins &

PeptidesInsulinCalcitonin

-Increased relative bioactivity in animals of 155.6% and 139.0% achieved

-DPI formulations showed promising in vitro deposition

Our research achievements Our research achievements continuedcontinued……

3.

Enhanced Pulmonary Absorption of Peptides

Our research achievements Our research achievements continuedcontinued……

ContraceptivesLevonorgesterolLeuprolide

100% 100% MaleMale and Female and Female contraception contraception achieved in rats.achieved in rats.

Our research achievementsOur research achievements continuedcontinued……

4.

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Mean Plasma level of Levonorgestrel in rats

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LOPlain drugMixture of constituentsLiposomes

Our research achievementsOur research achievements continuedcontinued……

Cystic Fibrosis

Amikacin sulphate and Tobramycin sulphate

-Spray dried Porous and Lyophillized Light and Large particles developed- In vivo evaluation indemonstratedprolonged drug levels in lung in rats

Our research achievementsOur research achievements continuedcontinued……5.

ImmunosupressionAfter lung

transplantationTacrolimus

-Nanoliposomal DPI with trehalose-In vivo evaluation in rats demonstrated prolonged drug levels in lung.

Our research achievementsOur research achievements continuedcontinued……6.

Pneumocystiscarinii

pneumonia

(PCP)

Dapsone

-Nanoliposomal DPI with hydrolized gelatin

In vitro release upto 24 hrs in Phosphate buffer pH 7.4 saline observed.

Our research achievementsOur research achievements continuedcontinued……7.

Cystic Fibrosis,&

Lung CancerGene Delivery

-Incarporation of genes (CFTR , p53 ) in liposomes for intranucleartargeting

Ongoing ..Ongoing ..

FEW CASE STUDIES FROM LITERATURE :

LIPOSOMALDPI BY

PULMONARY ROUTE

Opoid Analgesics

-Local as well as systemic analgesia, better than solution administered parentrally (iv, im) and oral.

[Ref] Bystrom, K., Nilsson, P.: US20001RE38407 (2001).

LIPOSOMALDPI BY

PULMONARY ROUTE

CorticosteroidalDerivatives

• Prolonged steroid retention in respiratory tract, improved therapeutic ratio, lower toxicity, reduced systemic side effects, and stability for several months

• Interstitial Diseases of Lung

[REF] Radhakrishnan, R.: US5049389 (1991).

FEW CASE STUDIES FROM LITERATURE :

NANOCOCHL-EATESDPI BY

PULMONARY ROUTE

Nanocochleates

-Liposome + Polymer solution treated with Ca++ or Zn++, yields precipitate of Nanocochleates (nm)

- Achieve efficient systemic and mucosal delivery

[REF] Parmar,M.: US20060051406A1 (2006).

FEW CASE STUDIES FROM LITERATURE :

AntibacterialAgents

Proliposomes

- Comprises Lipid of suitable Tg and antibacterial agent.

- On inhalation the drug spontaneously encapsulates into lipid inside lungs.

- Useful for Anthrax Therapy

[REF] Weers,J.G., Tarara, T., Tzannis, S.: US20050214224A1 (2005).

FEW CASE STUDIES FROM LITERATURE :

Research Publications

200115-27223Int. J. Pharm.DPI of liposomal Ketotifen Fumarate: Formulation and Characterization

Mayank Joshi, Ambikanandan Misra

1.

2001Article 25

2 (4)AAPS PharmSciTech

Liposomal Budesonide for Dry Powder Inhalation: Formulation & Stabilization

Mayank Joshi, Ambikanandan Misra

2.

2001531-53623 (10)Methods Find Exp Clin Pharmacol

Pulmonary disposition of Budesonidefrom Liposomal Dry Powder Inhaler

Mayank Joshi, Ambikanandan Misra

3.

2003153- 15630 (3)Clin. & Exp. Pharmacol. & Physiol.

Disposition kinetics of liposomal dry powder inhaler of Ketotifen fumaratein rats

Mayank Joshi, Ambikanandan Misra

4.

2003127-138 4 (2)AAPS PharmSciTech

Influence on Absorption Promoters on Pulmonary Insulin Bioactivity

T.Mahesh Kumar, Ambikanandan Misra

5.

2004140-1475(1) AAPS PharmSciTech

Pulmonary Absorption of Liposomal Levonorgestrel

Ambikanandan Misra and AliasgarShahiwala

6.

2004247-25311(4)

Drug delivery Development of Liposomal Amphotericin B Dry Powder Inhaler Formulation

Shah S.P., Misra Ambikanandan,

7

2004Article 65

5 (4)AAPS PharmSciTech

Liposomal Amikacin Dry Powder Inhaler: Effect of Fines on In Vitro Performance

Shah S.P., Misra Ambikanandan,

8

INTERNATIONAL

Year Pages Vol.(No)

Name of journalTitle of paper AuthorsS.No.

2007In Press3Journal of Biomedical Nanotechnology

Assessment of in-vitro antiproliferative activity of WGA-conjugated budesonide nanoparticles in A-549 cells

Naazneen Surti, Sachin Naik, and Ambikanandan Misra

16

2006465-47511(4)Pharmaceutical Development Technology

Optimization of formulation components and characterization of large respirable powders containing high therapeutic payload

Ambikanandan Misra, M. B. Chougule, B.K. Padhi

15.

2006677-68632 (6)Drug Development and Industrial pharmacy

Formulation and Evaluation of Insulin Dry Powder for Inhalation

T. Mahesh Kumar, Ambikanandan Misra

14.

20071-142(4)International Journal of Nanomedicine

Preparation, characterization, and pharmacokinetics of nano-liposomaldry powder inhaler of Tacrolimus

Ambikanandan Misra, M. B. Chougule, B.K. Padhi

13

20063001-3009

6 (9-10)Journal of Nanoscience and Nanotechnology

Nano-liposomal Dry Powder Inhaler of Amiloride Hydrochloride

Ambikanandan Misra, M. B. Chougule, B.K. Padhi

12

2005E-482-486

6 (3)AAPS PharmSciTechA preliminary pharmacokinetic study of liposomal leuprolide dry powder inhaler: a technical note.

Aliasgar Shahiwala , Ambikanandan Misra

11.

2004812-81359 (10)PharmazieLiposomal Amphotericin B dry powder inhaler: Effect of fines on in vitro performance

Shah S.P., Misra Ambikanandan,

10.

2004135-14412 (3)Journal of Drug Targeting

Pulmonary Absorption Enhancement of Salmon Calcitonin

T. MaheshKumar, Ambikanandan Misra

9.

Year Pages Vol.(No)

Name of journalTitle of paper AuthorsS.No.

2003336-34565(4)Indian J. Pharm. Sci.

Factors Affecting Development of Dry Powder Inhalers

S.P. Shah. S. Ganeshand A.N. Misra

20

2007SeptPharmabizDry Powder Inhaler: Novel approaches

Ambikanandan Misra21

200711-211Recent Patents on Drug Delivery & Formulation,

Development of Dry Powder Inhalers Ambikanandan Misra, M. B. Chougule, B.K. Padhi, K.A. Jinturkar

22

1999245-25236 (4)Indian DrugsLiposomes of TerbutalineSulphate:Preparation, Optimization and Stability Studies

Mayank Joshi, Ambikanandan Misra

17

1999881-88737 (9)Ind. J. Exp. Biol.Liposomes of terbutaline sulphate: In vitro and in vivo studies

Mayank Joshi, Ambikanandan Misra

18

200092-9662Indian J. Pharm. Sci.

Spectrophotometric determination of Budesonide

Mayank Joshi, Ambikanandan Misra

19.

REVIEWS AND GENERAL ARTICLES

NATIONAL

Year Pages Vol.(No)

Name of journalTitle of paper AuthorsS.No.

953/MUM/2006

30th June, 2006

Aerodynamically light porous dry powder inhalerformulations for targeted pulmonary deposition

Ambikanandan Misra,Mahavir BhupalChougule, S. Ganesh, Bijay Kumar Padhi.

3

729/MUM/2005

27th May 2005Enhancement of Pulmonary Therapeutic Index of Drugs from Dry Powder Inhaler Formulations.

Ambikanandan Misra, Mahavir Bhupal Chougule, Bijay Kumar Padhi,

2

228/MUM/2005

25th Feb, 2005Engineered Monodisperse Inhalation Powders for Effective Treatment of Lung Diseases.

Ambikanandan Misra,Bijay Kumar Padhi, Mahavir Bhupal Chougule

1

Patent office no.

Date of filing Tile of patent Inventors Sr.No.

PATENTS FILED:

Book publishedMisra A.N., “Advances In Pulmonary Drug Delivery” in “Advances in Controlled & Novel Drug Delivery (N.K. Jain Eds.)”, CBS Publishers & Distributors, New Delhi, pp 120-165 (2001).

Future Research Focus

Cell Culture StudiesCellular uptake using Calu- 3, Alveolar A-549 , MAC-7 Epithelial monolayer, MS-H macrophagecell line.Permeation through alveolar monolayer (Paracellular Transport, Transcellular Transport)Cellular and Molecular StudiesAnimal Studies and clinical evaluationPilot Scale up of Formulations

The pulmonary liposomal drug delivery has been very advantageous for prolonged and cell specific delivery of drugs. The recent advances in the post-genomic era and molecular biology, has improved it further. However, intracellular targeted therapy of therapeutic (specially biological) molecules using liposomal carrier with high safety and efficacy is the need of hour.

CONCLUSION

Thank You!