The role of dissolution testing in complementary and African traditional medicines: development of methods and applications

Isadore (Izzy) Kanfer

RHODES UNIVERSITY Where leaders learn

DISSOLUTION WORKSHOP CHALLENGES IN DISSOLUTION TESTING: EQUIVALENCE AND SURROGATES 9 -10 DECEMBER 2009 Barratt Lecture Complex, Rhodes University, Grahamstown South Africa,

Dissolution Testing

A QC Tool or Bioequivalence Predictor ?

Dissolution
This test is used to measure the release of an active substance (usually single ingredient) from the product in solid oral, tablet or capsule dosage forms.

It is one of the most important and useful in vitro tests for assuring product quality

In vitro dissolution often aids in guiding the selection of prototype formulations It helps to determine optimum amounts of ingredients needed to achieve requisite drug release profiles It is one of the most important and useful in vitro tests for assuring product quality batch-batch consistency Provides information on the impact of changes in composition, process or site of manufacture Can help identify potential problems of In vivo release and bioavailability/absorption following administration

In vitro dissolution is a valuable tool to assess drug product stability and shelf-life As products age, the formulations properties may change and alter the dissolution characteristics of the product over time For example: 1) moisture levels may increase or decrease over time and this can result in altered tablet hardness and subsequent possible changes in drug release rates 2) Polymorphic transformations affect solubility and dissolution

Disintegration

Deaggregation

Dissolution

Drug in Blood

Factors Affecting Dissolution Rate

Over the past 30 years or so, dissolution testing is probably the most important in vitro test that can be used to assess and control variables associated with formulation excipients, design and manufacturing which may alter the release characteristics of the active moiety from the dosage form.

In vitro

In vivo

L Shargel and ABC Yu, Applied Biopharmaceutics & Pharmacokinetics, 4th Edition, 1999

Banker et al, Lieberman and Lachman (eds)Pharmaceutical Dosage Forms: Tablets Volume1, 1980

Banker et al, Lieberman and Lachman (eds)Pharmaceutical Dosage Forms: Tablets Volume1, 1980

Guidelines For Dissolution Testing of Oral Dosage Forms (Orthodox Medicines) Developed USA FDA
(Guidance for industry: Dissolution testing of immediate release solid oral dosage forms. U.S. Dept of Health and Human Services, Food & Drug Administration, Center for Drug Evaluation and research (CDER), Rockville, USA, 1977)

FDA recommends use of buffers in the range 1.2 7.5 for dissolution media and i iti l method d di l ti di d initial th d development l t

Unlike orthodox medicines, specific guidelines for dissolution testing of complementary/alternate (CAMs) and traditional medicines (TMs) have not been developed

Also, there are no dissolution testing requirements for the quality control of such products. q y p

Application of dissolution testing as a quality control tool in complementary & alternative medicines (CAMs) and traditional medicines (TMs) ( )
CAMs/TMs, unlike conventional pharmaceutical products, which usually consist of one or two well- characterized active ingredients, are complex, consisting of multiple components that may be active individually or in combination What should be measured ? i.e. in contrast to chemically defined components , the actives in CAMs/TMs are generally defined to be the whole herbal preparation, e.g. the extract in its entirety

Chemical composition and classification of active pharmaceutical ingredients (APIs) in CAMs/TMs

Suggested that since the whole herbal product, e.g. extract, is regarded as the API, several extract types, depending on the p , yp , p g pharmaceuticalanalytical, pharmacological-toxicological and clinical findings, can be identified:

A B1

extracts containing constituents (single or groups) that are solely responsible for the activity (standardized extracts) extracts containing chemically defined constituents (single or g y ( g groups) known to contribute to pharmacological or synergistic activity (quantified extracts) extracts containing no constituents documented as being determinant or relevant for efficacy, or having pharmacological or clinical relevance.

B2

unknown active ingredients for which no individual active ingredients have been identified but have a traditional place in therapy of certain diseases. Use of chemical markers which may not contribute to any activity, e.g. Valerian

CASE STUDY
St Johns Wort Products Components in St Johns Wort known, or suspected, to play a role in its antidepressant activity include phloroglucinols, naphthodianthrones and flavonoids hence B1 category flavonoids, Hyperforin is purported to contribute to the antidepressive activity

Roles of other components in St Johns Wort ?

e.g.Naphthodianthrones (medium polarity)? Flavonoids (hydrophilic)? CONTENT UNIFORMITY and COMPLIANCE WITH THE LABEL CLAIM are standard quality criteria that need to be tested! What about dissolution testing? 5 St Johns Wort products were selected by Westerhoff et al. (2002), J.Pharm Ph (2002) J Ph Pharmacol., 54 1615 1621 l 54: 1615-1621 Dissolution of hyperforin, naphthodianthrones (hypericin & pseudohypericin) and flavonoids (isoquercitrin, quercitrin, rutin and hyperoside) were characterized to cover the entire spectrum of polarities present in the products.

FaSSIF=Fasted state simulated intestinal fluid; pH 6.5 FeSSIF=Fed state simulated intestinal fluid; pH 5.0 SGF=Simulated gastric fluid; pH 1.5
Westerhoff et al; (2002), J Pharm Pharmacol; 54; 1615-1621

Westerhoff et al; (2002), J Pharm Pharmacol; 54; 1615-1621

Biflavonoid Purported to be pharmacologically active.

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Coated tablet (St.Johns Wort) Tap water ?

N.B. Phenolic glycosides better dissolved than flavone aglycones

Jrgenliemk and Nahrstedt

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Pharmacological synergism and/or Physicochemical synergism?

Flavonoids (other included components) increase dissolution of hypericin ? What about synergistic effects on bioavailability ?

Pharmaceutical quality of different Ginkgo biloba brands Kressmann et al; J Pharm Pharmacol; 54: 661-669 (2002)

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Consider African Potato (AP):

a commonly used African Traditional Medicine (ATM) Apart from its perceived nutritional value used for: Immuno-boosting properties urinary diseases prostrate hypertrophy cancer ?

In South Africa African Potato gained prominence as an alternative medicine for nutritional use in the daily diet of HIV/AIDS patients recommended by the previous South African Minister of Health

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AP contains as its most important component, a norlignan di-glucoside, namely hypoxoside

It is also purported to contain a sterol, -sitosterol (BSS)

Currently, assay methods for quality control of AP products include:

H Hypoxoside (HPLC UV1 and CZE2) id (HPLCUV d
1

Nair & Kanfer, J Agric Food Chem, 54:2816-2821,2006 & Kanfer, Phytochem Anal, 18:475-483, 2007

2 Nair

Stigmasterol, -sitosterol and stigmastanol (HPLC with evaporative light-scattering detection3)

3

Nair , Kanfer & Hoogmartens, J Pharm Biomed Anal, 41:731-737,2006

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Approaches used for the development of a suitable dissolution method for products containing AP:
Phytochemical composition? - Hypoxoside - Sterols & sterolins but which?

First step = Assay!

Assay results:
-sitosterol major component Hypoxoside Used as markers for Quality Control

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List of AP Products:

Initial dissolution tests:

Products A,B,D,E,F & G - all capsules USP-1 (Basket) P d t C - th only t bl t d Product the l tablet dosage f form - USP 2 (P ddl ) USP-2 (Paddle) Dissolution Conditions: Volume 900 ml Rotation speed 100 rpm p p Temperature 37 0.5 C Dissolution media as per USA FDA Guidance buffered dissolution media (pH 1.2 7.5)

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 Hypoxoside relatively unstable at higher pHs, hence used acidic dissolution media only, i.e. pH 1.2 and 4.5 Product B used since it contained highest content of hypoxoside

-sitosterol relatively insoluble in aqueous media Dissolution performed at pHs 1.2, 4.5, 6.8 and 7.8 NO -sitosterol detected even after 12 hours i.e i e confirmation of aqueous insolubility Choice of alternate dissolution media necessary FaSSIF (fasted state simulation - generally pH 6.5) FeSSIF (fed state simulation - generally p 5.0) ( g y pH )

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Dissolution of product D for BSS in FaSSIF & FeSSIF at various pHs

Dissolution comparison of Products A, B and C at pH 1.2

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Dissolution profiles of Products D, E, F & G showing profiles of BSS at pH 5.0 in FeSSIF

Issues with African Traditional Medicines (ATMs) Sceletium Species - example

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Reference substances: Example Sceletium Alkaloids

OCH3
4' 5' 6' 1' 3 2 3a 3' 2' 4 5 6

OCH3
OCH3

OCH3 OCH3 OCH3

OCH3 OCH3

HCl

N CH3

7a

N CH3

N CH3

N CH3

Mesembrine (Major alkaloid)

7Mesembrenone

Mesembrenone

Mesembrine hydrochloride crystals

OCH3 OCH3

OCH3 OCH3

OCH3 OCH3

N CH3

OH H

N CH3

OH H

N CH3

OH H

Mesembranol crystals

Epimesembranol

Mesembrenol

Dissolution profile of Sceletium capsules in Buffer pH 6.5

MeanP rofiles
AP 0001/09 T es tP roduc t
120

S c eletiumC aps ule

NNNNN

100

80 % R eleas ed

60

40

20

0 0 50 100 150 T im e (Hours) 200 250 300

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Summary
Complexity of CAMS/TMs Role of dissolution as a quality control tool for CAMS/TMs What to monitor? Bio-relevance of dissolution testing ?

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