pION Instruments Flux Measurements using Pion …...Fiber Optics Advanced Training Course | May 24 – 25, 2016| Pion Inc. Billerica MA, USA Number of Permeability Chambers: 4 donor/receiver

Fiber Optics Advanced Training Course | May 24 – 25, 2016| Pion Inc. Billerica MA, USA

pION Instruments Flux Measurements using Pion µFLUX


Solubility/Permeability in Absorption Biopharmaceutics Classification System (BCS)

HIGH PERMEABILITY

LOW PERMEABILITY

HIGH SOLUBILITY LOW SOLUBILITY

a RATE OF DISSOLUTION limits in vivo absorption b SOLUBILITY limits absorption flux c PERMEABILITY is rate determining d No IVIV (in vitro - in vivo) correlation expected

1 2 CLASS 1 (some uptake?) a diltiazem antipyrine labetolol glucose captopril L-dopa enalapril metoprolol propranolol phenylalanine 3 4 CLASS 3 (hydrophilic) c famotidine atenolol cimetidine acyclovir ranitidine nadolol hydrochlorothiazide

CLASS 2 (lipophilic) b flurbiprofen ketoprofen naproxen desipramine diclofenac itraconazole piroxicam carbamazepine phenytoin verapamil

CLASS 4 (some efflux?)d terfenedine furosemide cyclosporine

www.fda.gov/cder/guidance/2062dft.pdf

pH 1 – 7.5


Differential equations for oral drug absorption

Dissolution Permeation

API particles Molecules

Sugano, K. Expert Opin Drug Metab Toxicol 2009, 259-293

dissolvpermperm Xk

dtdX

⋅=

⋅

−⋅−≈GIdissolv

dissolvdiss

solid

VSXDosek

dtdX 1

Solutions？


Absorption Limited Processes

Solid

Dissolved

Absorbed

Xsolid

Xsolu

Xabs

(C) Solubility Limited

Dissolved

Solid

Absorbed

Xsolid

Xabs

(B) Permeability Rate Limited

Dissolved

Absorbed

Dissolution rate Solubility

Permeation rate

Solid

Xsolid

Xsolu

Xabs

(A) Dissolution Rate Limited

(Sbulk – Cbulk(t))

(A) Dissolution rate limited absorption (Rate of Dissolution < Rate of Permeation) (B) Permeability limited absorption (C) Solubility limited absorption (Amount of Available Material Limits Flux)

Courtesy of Kiyohiko Sugano; Sugano et al, DMPK 2007 (4) 225-254


Absorption Limiting Steps

Limiting Step Conditions Comments

Dissolution Tdiss > 199 min Peff > 2x10-4 cm/s

Dabs >> Dose

Mainly refers to particle size, absorbed amount non-

proportionally increases with dose

Permeability Tdiss < 50 min Peff < 2x10-4 cm/s

Dabs >> Dose

Highly soluble drugs or drugs dosed in solution, absorbed

amount proportionally increases with dose

Solubility Tdiss < 50 min Peff > 2x10-4 cm/s

Dabs < Dose

Saturation occurs, absorbed amount does not increases with

dose

L.X. Yu Pharm. Res. 1999, 16 (12) 1883


Flux Through Membranes

the net number of moles of particles

crossing unit area per unit time perpendicular to unit area

dtdc

AV

dtAdmFlux =⋅

=

Absorption Relates to Flux


Flux and Permeability

Kp = Cm0 / CD

Kp = Cmh / CA

Cm0

0 h

Cmh

CD

CA

x

C

dCm/dx = (Cm0 - Cm

h) / h

DONOR ACCEPTOR

FICK’S FIRST LAW

flux = Dm dCm/dx

= Dm [Cm0 - Cmh] / h

= Dm Kp (CD - CA) / h

= Pm (CD-CA) NHO

OH

OCH3 CH3

CH3

NHO

OH

OCH3 CH3

CH3

NHO

OH

OCH3 CH3

CH3

NHO

OH

OCH3 CH3

CH3

NHO

OH

OCH3 CH3

CH3

NHO

OH

OCH3 CH3

CH3

NHO

OH

OCH3 CH3

CH3

NHO

OH

OCH3 CH3

CH3

NHO

OH

OCH3 CH3

CH3

NHO

OH

OCH3 CH3

CH3

NHO

OH

OCH3 CH3

CH3

NHO

OH

OCH3 CH3

CH3

NHO

OH

OCH3 CH3

CH3

NHO

OH

OCH3 CH3

CH3

NHO

OH

OCH3 CH3

CH3

NHO

OH

OCH3 CH3

CH3

NHO

OH

OCH3 CH3

CH3 NHO

OH

OCH3 CH3

CH3

NHO

OH

OCH3 CH3

CH3

NHO

OH

OCH3 CH3

CH3

NHO

OH

OCH3 CH3

CH3

NHO

OH

OCH3 CH3

CH3

NHO

OH

OCH3 CH3

CH3

NHO

OH

OCH3 CH3

CH3

Fiber Optics Advanced Training Course | May 24 – 25, 2016| Pion Inc. Billerica MA, USA Copyright © Pion Inc. 2015

In Vitro Permeability Setup: no sink conditions, Aqueous Boundary Layer

0 h x

C

DONOR ACCEPTOR

NHO

OH

OCH3 CH3

CH3

NHO

OH

OCH3 CH3

CH3

NHO

OH

OCH3 CH3

CH3

NHO

OH

OCH3 CH3

CH3

NHO

OH

OCH3 CH3

CH3

NHO

OH

OCH3 CH3

CH3

NHO

OH

OCH3 CH3

CH3

NHO

OH

OCH3 CH3

CH3

NHO

OH

OCH3 CH3

CH3

NHO

OH

OCH3 CH3

CH3

NHO

OH

OCH3 CH3

CH3

NHO

OH

OCH3 CH3

CH3

NHO

OH

OCH3 CH3

CH3

NHO

OH

OCH3 CH3

CH3

NHO

OH

OCH3 CH3

CH3

NHO

OH

OCH3 CH3

CH3

NHO

OH

OCH3 CH3

CH3

NHO

OH

OCH3 CH3

CH3

NHO

OH

OCH3 CH3

CH3

NHO

OH

OCH3 CH3

CH3

NHO

OH

OCH3 CH3

CH3

NHO

OH

OCH3 CH3

CH3

NHO

OH

OCH3 CH3

CH3

NHO

OH

OCH3 CH3

CH3

NHO

OH

OCH3 CH3

CH3

NHO

OH

OCH3 CH3

CH3

NHO

OH

OCH3 CH3

CH3

NHO

OH

OCH3 CH3

CH3

NHO

OH

OCH3 CH3

CH3

ABLDON

ABLACC

hABL hABL


Diffusional Resistance of Series is Additive

mABLe PPP111

+=

Depends on stirring or agitation efficiency

and size of the molecule

Effective measured permeability

membrane permeability


mABLe PPP111

+=

ABLm PP >> ABLe PP ≈

ABLm PP <<me PP ≈

1)

2)

Limiting Cases


m

mmm h

DkP ⋅=

What is PABL or PUWL

ABL

aqABL h

DP =


Permeability through membranes combines partitioning and diffusion does not depend on concentration

FLUX depends on dissolution rate, solubility and permeability

m

mmm h

DkP =

)(tcPdtdc

AVFlux De ⋅==

Flux: dissolution/solubility/permeability

ABL

mme

PPPP

+=

1

1


Determined by FLUX of the API through membranes

)(tcPdtdc

AVFlux De ⋅==

Absorption Phase of PK Profile

Avdeef A, Kansy M., Bendels S., Tsinman K. Absorption-excipient-pH classification gradient maps: Sparingly soluble drugs and the pH partition hypothesis. Eur. J. Pharm. Sci. 2008, 33, 29 – 41.

Dahan A., Miller J.M. The Solubility-Permeability Interplay and Its Implications in Formulation Design and Development for Poorly Soluble Drugs. The AAPS J. (2012), Vol. 14, No. 2, 244-251

ABLme PPP111

+=

プレゼンター

プレゼンテーションのノート

Fick’s first law connects flux with


Why Dissolution Study May Not Predict In Vivo Data?

M. Kataoka, et al., Pharm. Res., 2012, 29, 1485-1494


What Limits the Absorption µFLUX™ Apparatus

)()( tcPdtA

dmtJFlux e ⋅=⋅

=

Combining benefits of in situ concentration monitoring with dissolution-permeability setup

Membrane Holder

Dissolution-Permeation Glass Pair


Number of Permeability Chambers: 4 donor/receiver pairs mounted on MB8 platform

Working Volume: 16 – 22 mL

Area of Membrane Holders (Teflon): 14 mm (1.54 cm2)

Stirring: Magnetic stir bars, individual computer RPM control for all

chambers

Permeation Barriers: a) Filter supported artificial membranes (PAMPA)

b) Filter supported cell monolayers (Caco-2, MDCK, etc.) c) Size exclusion membranes (dialysis membranes)

d) Skin or skin sub-layers layers (same as Franz cell systems) e) User designed

Temperature Control: Connection to external water bath

µFLUX™ Specifications (Rev. 2)


About Membrane: Double-Sink™ PAMPA correlates with GIT

Avdeef et al. J Pharm Sci. 2007 Nov;96(11):2893-909.


µFLUX Permeability Experiment Carbamazepine

Acceptor:

Acceptor Sink Buffer (ASB, pH 7.4)

Donor:

Prisma™ HT buffer, pH 6.5 Initial concentration: 70 µg/mL

Membrane:

PVDF support (hydrophobic, 0.45 µm pore size) GIT lipid (Pion Double-Sink™ Model)

Permeability Calculation:

Slope*Vol/Area/cD(0) ~ 84*10-6 cm/s

Acceptor Compartment

Donor Compartment


Solubility Enhancement ~ 42 times

Formulated API ~ 250 µg/mL after 10 min

Unformulated API ~ 6 µg/mL after 60 min

Flux 0.939 µg/(min*cm2)


Flux Enhancement ~ 40 times

Case Study #1: Enhanced FLUX


Solubility Enhancement ~ 2.5 times

Flux Suppression ~ 1.6 times

Case Study #2: Suppressed FLUX

Formulated API ~ 950 µg/mL after 0.1 min

Unformulated API ~ 400 µg/mL after 30 min




Research Compound FLUX

Donor: Load 67 µg/mL SIF: 0.2 % Fast Conditions

µFLUX Experiment: Studying Food Effect

Donor: Load 67 µg/mL SIF: 2 % Fed Conditions

Flux: 5.9*10-2 µg/(min cm2)


Negative Food Effect Found in Animal Studies

500% Solubility 10% Flux Increase


Research Compound FLUX


µFLUX Experiment: Dose Effect




5x DOSE

Solubility Limited Flux


µFLUX Experiment Danazol in Buffer (FeSSIFBlank pH 5.0)


FLUX in FeSSIFBlank

1.0*10-4 µg/(cm2 s)

Permeability of Danazol

~411*10-6 cm/s


µFLUX Experiment Danazol in FeSSIF (Food effect)

Donor Compartment


13 µg/mL

Solubility Enhancement > 40 fold

Flux Enhancement ~ 5.4 fold

FLUX in FeSSIF

5.4*10-4 µg/(cm2 s)

Permeability of Danazol in FeSSIF

~42*10-6 cm/s

Human Food Effect: ~4 fold


0

5

10

15

20

25

30

35

0 100 200 300 400 500 600 700 800 900 1000

Conc

entr

atio

n (u

g/m

L)

Time (minutes)

Danazol: Dissolution profiles in the donor chambers

in FaSSIF-full & FeSSIF-full media at 0.4mg/mL

Danazol_FeSSIF Full-Rep1Danazol_FeSSIF Full-Rep2

-10.0

0.0

10.0

20.0

30.0

40.0

50.0

0 100 200 300 400 500 600 700 800 900 1000

Conc

entr

atio

n (u

g/m

L)

Time (minutes)

Danazol: Appearance profiles in the acceptor chambers for corresponding

donors in FaSSIF-Full & FeSSIF-Full medium.

Danazol_FeSSIF Full -Rep1Danazol_FeSSIF Full -Rep2Danazol_FaSSIF Full-Rep1

µFLUX: Positive Food Effect (~5 fold)

Human Food Effect: ~4 fold

µFLUX Experiment Food Effect Danazol in FaSSIF vs FeSSIF


µFLUX Experiment Food Effect Gresiofulvin in FaSSIF vs FeSSIF

0

5

10

15

20

25

30

35

40

0 200 400 600 800 1000

Conc

entr

atio

n (u

g/m

L)

Time (minutes)

Griseofulvin: Dissolution profiles in the donor chambers

in FaSSIF-full & FeSSIF-full media at 0.6mg/mL

Griseofulvin_FaSSIF Full-Rep1

Griseofulvin_FaSSIF Full-Rep2

Griseofulvin_FeSSIF Full -Rep1

Griseofulvin_FeSSIF Full-Rep2

-1.0

4.0

9.0

14.0

19.0

24.0

0 100 200 300 400 500 600 700 800 900 1000

Conc

entr

atio

n (u

g/m

L)

Time (minutes)

Griseofulvin: Appearance profiles in the acceptor chambers for

corresponding donors in FaSSIF-Full & FeSSIF-Full medium.

Griseofulvin_FaSSIF Full -Rep1Griseofulvin_FaSSIF Full -Rep2

µFLUX: No Food Effect

Human: No effect or slightly positive


Nano ppt like turbidity, saturated

spectrum

PRZ pH 6.5 PRZ pH 6.5 PRZ pH 7.4 PRZ pH 7.4

PRZ pH 7.4

ZIM shift

ZIM shift

PRZ pH 8.5 PRZ pH 8.5 PRZ pH 8.5

No shift

ZIM shift

No shift

No shift

No shift No shift

No shift Supersaturation

Peculiar Supersaturation of Prazosin.HCl

pH 6.5

pH 6.5 pH 7.4

pH 7.4

pH 7.4 pH 8.5 pH 8.5

pH 8.5

Load 200 µg/mL


Flux of Prazosin.HCl at Different Loads

pH 6.5


Raina S.A., et al. Enhancements and Limits in Drug Membrane Transport Using Supersaturated Solutions of Poorly Water-

Soluble Drugs. DOI 10.1002/jps.23826

Amorphous Solubility Limits Flux

Joint Research of Abbvie and Purdue University Scientists

Felodipine 25 C

Felodipine 37 C

Nifedipine 37 C


Dissolution/Flux in FaSSIF

Research Compound SGF – FaSSIF transformation

after 30 min of the assay

Winner formulation performed best in dog model


Amorphous Solid Dispersions of Meloxicam: Flux

Untreated Meloxicam API Load 105 μg/mL Soluplus Formulation

API Load 134 μg/mL

VA 64 Formulation API Load 108 μg/mL

VA 64/TPGS Formulation API Load 121 μg/mL

FLUX FLUX

FLUX FLUX


How to make sense of complex supersaturating formulations

Flux is not constant!

Area under Concentration-Time Profile in Receiver as

Formulation Ranking Parameter


… “findings [of the study] clearly point out the flaws in using solute concentration in estimating solute activity or supersaturation, and reaffirm the use of flux measurements to understand supersaturated systems.”

Citation

Acknowledgements

Lab Team

Oksana Tsinman Ram Lingamaneni Janki Patel


Fiber Optic Validation Publications

1. Liu L., Fitzgerald G., Embry M., Cantu R., and Pack B. Technical Evaluation of a Fiber-

Optic Probe Dissolution System. Dissolution Technol. 2008, 15 (1), 10-20

2. Bynum K., Roinestad K., Kassis A., Pocreva J., Gehrlein L., Cheng F., and Palermo P. Analytical Performance of a Fiber Optic Probe Dissolution System. Dissolution Technol. 2001, 8 (4), 13-21

3. Toher C., Nielsen P., Foreman A., and Avdeef A. In Situ Fiber Optic Dissolution Monitoring of a Vitamin B12 Solid Dosage Formulation. Dissolution Technol. 2003, 10 (4), 20-25

4. Zolnik B.S., Raton J-L., and Burgess D.J. Application of USP Apparatus 4 and In Situ Fiber Optic Analysis to Microsphere Release Testing. Dissolution Technol. 2005, 12 (2), 11-14

5. Bijlani V., Yuonayel D., Katpally S., Chukwumezie B.N., and Adeyeye M.C. Monitoring Ibuprofen Release from Multiparticles: In Situ Fiber-Optic Technique versus the HPLC Method: A Technical Note. AAPS PharmSciTech 2007, 8 (3), E1-E4


µDISS Profiler Publications (incomplete)

1. Tsinman K., Avdeef A., Tsinman O., and Voloboy D. Powder Dissolution Method for Estimating Rotating Disk Intrinsic Dissolution Rates of Low Soluble Drugs. Pharm. Res. 2009, 26 (9), 2093-2100

2. Avdeef A., Tsinman O., Miniaturized Rotating Disk Intrinsic Dissolution Rate Measurement: Effect of Buffer Capacity in Comparison with Traditional Wood’s Apparatus. Pharm. Res. 2008, 25 (11), 2613-2627.

3. Avdeef A., Tsinman K., Tsinman O., Sun N., and Voloboy D. Miniaturization of Powder Dissolution Measurement and Estimation of Particle Size. Chem. Biodiv. 2009, 6 (11), 1796-1811

4. Fagerberg J.H., Tsinman O., Sun N., Tsinman K., Avdeef A., and Bergström C.A.S. Dissolution Rate and Apparent Solubility of Poorly Soluble Drugs in Biorelevant Dissolution Media. Mol. Pharm. 2010, 7 (5), 1419-1430

5. Alonzo D.E., Zhang G. G. Z., Zhou D., Gao Y., and Taylor L.S. Understanding the Behavior of Amorphous Pharmaceutical Systems during Dissolution. Pharm. Res. 2010, 27 (4), 608-617

6. Furukawa S., Zhao C, and Ohki Y. Methodology for phase selection of a weak basic drug candidate, utilizing kinetic solubility profiles in bio-relevant media. Eur. J. Pharm.&Biopharm. 2010, 74, 298 – 303.

7. Nielsen L. H., et al. Preparation of an amorphous sodium furosemide salt improves solubility and dissolution rate and leads to a faster Tmax after oral dosing to rats. Eur. J. Pharm.&Biopharm. http://dx.doi.org/10.1016/j.ejpb.2013.09.002

8. Fagerberg J.H., et al. Ethanol Effects on Apparent Solubility of Poorly Soluble Drugs in Simulated Intestinal Fluid. Mol. Pharm. 2012, 9, 1942−1952.

9. Mathias N., et. al. Assessing the Risk of pH-Dependent Absorption for New Molecular Entities: A Novel in Vitro Dissolution Test, Physicochemical Analysis, and Risk Assessment Strategy. Mol. Pharm. 2013, 10, 4063 – 4073.

10. Kogermann K., et. al. Dissolution testing of amorphous solid dispersions. Int. J. Pharm. 2013, 444, 40 – 46. 11. Nielsen L.H., et. al. Biorelevant characterization of amorphous furosemide salt exhibits conversion to a furosemide

hydrate during dissolution. Int. J. Pharm. 2013, 457, 14 – 24. 12. Martin F.A., et. al. Ketoconazole Salt and Co-crystals with Enhanced Aqueous Solubility. Cryst. Growth Des. 2013, 13,

4295 – 4304. 13. Fagerberg J.H., et. al. Concomitant intake of alcohol may increase the absorption of poorly soluble drugs. Eur. J.

Pharm. Sci. 2015, 67, 12 – 20.

http://dx.doi.org/10.1016/j.ejpb.2013.09.002

Documents

pION Instruments Flux Measurements using Pion …...Fiber Optics Advanced Training Course | May 24 – 25, 2016| Pion Inc. Billerica MA, USA Number of Permeability Chambers: 4 donor/receiver