BIO-RELEVANT DISSOLUTION MEDIA

INTRODUCTION
 Simulation of gastrointestinal conditions is essential to adequately predict the in vivo behavior of poorly soluble drugs.  Simulating small intestinal conditions with biorelevant media such as fasted state simulated intestinal fluid (FaSSIF) and fed state simulated intestinal fluid (FeSSIF) has become standard practice in many dissolution laboratories .

 However, due to their complex composition, these media are expensive and, to date, need to be prepared on the day of the experiment.  The aim of the present study was to develop media that are easier to prepare and are stable over a longer period, but can still serve the purpose of forecasting in vivo performance.  Criteria for developing simplified test media also include cost effectiveness and the ability to adequately reflect the physicochemical properties of the bio relevant media FaSSIF or FeSSIF .

 The aim of the present study was to develop media that are easier to prepare and are stable over a longer period, but can still serve the purpose of forecasting in vivo performance.  Criteria for developing simplified test media also include cost effectiveness and the ability to adequately reflect the physicochemical properties of the bio relevant media FaSSIF or FeSSIF

PREPARATION OF BIORELEVANT MEDIA

 MATERIALS Brij 35 (polyoxyethyleneglycol dodecyl ether) Span 80 (sorbitan monooleate) Tween 20 (polyoxyethylene sorbitan monolaurate) Tween 40 (polyoxyethylene sorbitan monopalmitate) Tween 60 (polyoxyethylene sorbitan monostearate) Tween 80 (polyoxyethylene sorbitan monooleate)

PREPARATION OF BLANK FaSSIF

Dissolve 1.74 g of NaOH (pellets), 19.77 g of NaH2PO4.H2O or 17.19 g of anhydrous NaH2PO4, and 30.93g of NaCl in 5 L of purified water.  Adjust the pH to exactly 6.5 using 1 N NaOH or 1 N HCl. 

PREPARATION OF FaSSIF
Dissolve 3.3 g of sodium taurocholate in 500 mL blank FaSSIF.  Add 11.8 mL of a solution containing 100 mg /ml lecithin in methylene chloride, forming an emulsion.  The methylene chloride is eliminated under vacuum at about 40C. Draw a vacuum for fifteen minutes at 250 mbar, followed by 15 minutes at 100 mbar.  This results in a clear,micellar solution, having no perceptible odor of methylene chloride.  After cooling to room temperature, adjust the volume to 2 L with blank FaSSIF. 

PREPARATION OF BLANK FeSSIF

Dissolve 20.2 g of NaOH (pellets), 43.25 g of glacial acetic acid, and 59.37 g of NaCl in 5 L of purified water  Adjust the pH to exactly 5.0 using 1 N NaOH or 1 N HCl. 

PREPARATION OF FeSSIF
Dissolve 16.5 g of sodium taurocholate in 500 mL of blank FeSSIF. Add 59.08 mL of a solution containing 100mg/mL lecithin in methylene chloride, forming an emulsion.  The methylene chloride is eliminated under vacuum at about 40C. Draw a vacuum for fifteen minutes at 250 mbar, followed by 15 minutes at 100 mbar.  This results in a clear to slightly hazy,micellar solution having no perceptible odor of methylene chloride.  After cooling to room temperature, adjust 

PHYSICAL CHEMICAL EVALUATION OF THE MEDIA

 The pH of all test media was measured with a pH meter.  The buffer capacity was quantified by potentiometric titration with 0.1 N hydrochloric acid.  The osmolality was measured by semi micro osmometry (osmometer type  Surface tension was determined with a bubble pressure tensiometer

PHYSICO CHEMICAL PARAMETERS OF THE TEST MEDIA: TARGET VALUES AND TOLERANCES SIMULATED FeSSIF SIMULATED FaSSIF  Parameter target value range of tolerance target value range of tolerance pH 6.5 0.1 5.0 0.1  Osmolality [mOsmol/kg] 270 15 670 15  Buffer capacity [mEq/pH/L] 12 2 72 2  Surface tension [mN/m] 54 2 48 2

BIO-RELEVANT MEDIUM
Simulated Gastric Fluid (SGF)
HCl (mL) NaCl (g) Deionized water 1.4 mL (conc. 12N HCl) 2.0 g (34.2 mM) 1L

Fasted Simulated Small Intestinal Fluid (FaSSIF)1

Na Taurocholate Lecithin KH2PO4 KCl NaOH Deionized water 1.61 g (3 mM) 0.56 g (0.75 mM) 3.9 g 7.7 g qs pH 6.5 1L

MEDIA PREPARATION
The biorelevant media FaSSIF and FeSSIF were prepared on the day of the experiment. FaSSIF was prepared as follows:  3.3 g sodium taurocholate was dissolved in approximately 500 mL of the blank FaSSIF.  The weight of this mixture was checked and noted (weight 1).  Then 11.8 mL of a methylene chloride solution containing 100 mg/mL lecithin (= 1.18 g lecithin, weight 2) was added.  This produced an emulsion (i.e., the resulting product was turbid). 

 The methylene chloride was then evaporated under vacuum using a Rotavap at a temperature of about 40 C. About 10 min at 500 mbar followed by 30 min at about 50 mbar led to complete removal of the methylene chloride.  The result was a clear, micellar solution having no perceptible odor of methylene chloride. After cooling to room temperature, the weight of the solution was checked again.  The water lost to evaporation was replaced with demineralized water to obtain a total weight corresponding to the sum of weight 1 and weight 2.  Finally, the volume was brought to 2 L with blank FaSSIF.

Likewise, FeSSIF was prepared by first dissolving 16.5 g sodium taurocholate in 500 mL of blank FeSSIF, checking and noting the weight (weight 1).  Subsequently, 59.1 Ml of a methylene chloride solution containing 100 mg/mL lecithin (= 5.91 g lecithin, weight 2) was added, resulting in an emulsion.  The methylene chloride was then evaporated under the conditions described for FaSSIF until a clear, micellar solution with no perceptible odor of methylene chloride was obtained.

After cooling to room temperature, the weight of the solution was checked again, and the water lost to evaporation was replaced with demineralized water to obtain a total weight corresponding to the sum of weight 1 and weight 2.  Finally, the volume was brought to 2 L with blank FeSSIF.  These methods of manufacturing FaSSIF and FeSSIF differed slightly from those proposed in the literature (11,12)

Simplified media containing different types and amount of surfactants were prepared using blank FaSSIF and FeSSIF (i.e., the relevant buffers without sodium taurocholate and lecithin) as the basis.  After the surfactant(s) were added, the media were placed in the ultrasonic bath for 15 min and then stirred for another 15 min on a magnetic stirrer.  Subsequently, various physicochemical parameters such as pH, surface tension, critical micelle concentration, osmolality, and buffer capacity were measured and compared with those of the biorelevant media.

ROLE OF BIORELEVANT MEDIA IN FORMULATION DEVELOPMENT

Challenges in early phase formulation development  Dissolution in Bio relevant media Simulated Gastric Fluid (SGF) and Fasted Simulated Small Intestinal Fluid (FaSSIF) Medium Simple two stage dissolution model (SGF followed by FaSSIF)for weak acids and weak bases  Five Merck case studies of using dissolution in bio relevant medium to guide formulation selection  Conclusions Dissolution in bio relevant medium can be a useful tool for early formulation selection

CHALLENGES IN EARLY PHASE FORMULATION DEVELOPMENT

Majority of the drug candidates have poor aqueous solubility  Early phase formulation needs to maximize bioavailability to ensure that API is delivered to the intended target  Variety of formulations options available  Limited API and limited formulation materials  In vitro dissolution, if reflecting in vivo performance, can be used to guide formulation development 

DESIRED DISSOLUTION FOR EARLY PHASE

Traditional Dissolution Desired Dissolution forVolume 500mL to 900mL Smaller volume medium Aqueous or surfactant Bio relevant medium: SGF, FaSSIF, medium with 3 10X sink FeSSIF  Profile Complete release in 60min Relative release rate, ranking order  Detection HPLC or UV (online, offline) Real time measurement  Focus on Bio relevant Method and  optimize formulation bioavailability  Focus on performance  checking (QC) method  Bio relevant media are designed to simulate gastrointestinal conditions, and

CASE 1: FORMULATION DEVELOPMENT FOR A LOW SOLUBLE NEUTRAL COMPOUND

 Compound A is a neutral compound. Aqueous solubility is < 0.01mg/mL in both SGF and FaSSIF  BCS II with high permeability  Various formulations were developed for Phase I LFC 1 3 with various lipid surfactant vehicles  Two Solid dosage forms with different type of surfactants  Bench mark of safety assessment formulation  Formulations are evaluated in bio relevant medium (FaSSIF)

CASE 2: BIO-RELEVANT DISSSOLUTION IS MORE DISCRIMINATING

Formulation 1 and 2 (filler type 1) have faster release than formulation 3(filler type in FaSSIF medium

In-vivo Animal PK result

Formulation Formulation-1 Formulation-3 AUC0-24hr ( M*hr) 82.1 69.1 Cmax ( M) 10.9 10.3

Formulation 1 has slight Higher bioavailability tha formuation 3,correlate w in vitro results in biorel medium

CASE STUDY 3: FORMULATION SELECTION WITH 2-STAGE BIO RELEVANT DISSOLUTION

 Dissolution was conducted in 90mL SGF and 180mL of FaSSIF with USP II paddle (100rpm)  The safety formulation (10% tween) remain supersaturated in SGF and Tween  Solid formulation A has the highest release in SGF and FaSSIF  In vivo animal study showed that solid

CASE STUDY 5: FORMULATION SELECTION IN FaSSIF MEDIUM

 Dissolution was conducted in 500mL of FaSSIF medium withUSP II paddle (100rpm  Formulation A and B release much faster than formulation C  In vivo animal study confirm the same ranking order observed invitro  Surfactant A was selected over

A SIMPLE DISSOLUTION MODEL SGF OR FaSSIF

INVIVO

Target Concentration= projected efficacious dose/volume of bio-relevant medium

BIO -RELEVANT MEDIUM VOLUME ADJUSTED BASED ON FORMULATION AVAILABILITY AND TARGET CONCENTRATION
Micro Dissolution USP II Paddle 200mL 1L

4-20 mL

Vessel Type Micro Dissolution Vankel 200mL Vankel 1000mL

Volume 4-20mL 50-200mL 250-1000mL

Agitation Stir bar Paddle Paddle

CONCLUSION
 Selected media containing synthetic surfactants with physicochemical properties similar to those of FaSSIF and FeSSIF were identified.  These can be used to replace biorelevant test media for screening formulations if it can be shown that solubility and dissolution of the drug in these simplified media are similar to the profiles in biorelevant media prepared with bile components.

BIBILOGRAPHY
 1. Dressman, J. B., Reppas, C. In vitro-in vivo correlations for lipophilic, poorly water-soluble drugs. B.T.Gattefosse, 93: 91100, 2000.  2. Nicolaides, E., Symillides,M., Dressman, J. B., Reppas, C. Biorelevant dissolution testing to predict the plasma of lipophilic drugs after oral administration. Pharm. Res., 18(3): 380388, 2001.  3. Horter,D.,Dressman, J. B. Influence of physicochemical properties on dissolution of drugs in the gastrointestinal tract. Adv. Drug Del. Rev., 46: 7587, 2001.  4. Lobenberg, R., Kramer, J., Shah,V. P., Amidon, G. L.,Dressman, J. B. Dissolution testing as a prognostic tool for oral drug absorption: Dissolution behavior of glibenclamide. Pharm.