Polymorphism in Pharmaceutical Drugs: Introduction

Silvia L. Cuffini
Agencia Crdoba Ciencia Unidad Ceprocor Crdoba - Argentina. Reseacher of CONICET

Outline
Definition / Properties of Polymorphism Relevance of Polymorphism: Res. ANVISA - FDA Studies of different examples:
Chlorpropamide: Methodology applied to study

polymorphism Progesterone: Impact of Polymorphism in design of intrauterine devices (Innovation transfered to a company) Mebendazole: Impact of Polymorphism in the Pharmaceutical Equivalence and Bioavalilability Patents and Polymorphism: Efavirenz Services and Facilities LRA-Sector DRX Conclusions

Polimorfismo: Solid

State Properties

Pharmaceutical Drugs: more than 80% are solid formulations Molecular Structure Crystal structure with order Crystalline Forms

Polymorph

Habits

Pseudo polymorph

Solvates Hydrates

Habits Habits

Structure without order

Amorphous

Solid State Properties: Polymorph

C atoms

Graphite

Diamond

Polymorphism: Definition

Polymorphism is the ability of a substance to exist in more than one crystal structure
Italian way!!

Solid State Properties: Polymorphs

Polymorphism is the ability of a substance to exist in more than one crystal structure
Polymorphs: when two crystals have the same chemical composition but different internal structure (molecular packing molecular conformation or / and inter or intra molecular interactions)

modifications or polymorphs or forms

Pseudo polymorphs : different crystal forms have molecules of the same given substances and also contain molecules of solvent incorporated into a unique structure (solvates or hydrates (water))
Byrn, Solid-state chemistry of drugs (1999)

Solid State Properties: Polymorphs Definicin

Mono-component systems: Polymorphs Multi-component systems

Walter C. McCrone 1916-2002

The number of crystalline forms or polymorphs that can be known of a compound are proportional to the time and resources dedicated to the investigation of them Mc Crone (1963)

Occurrence of Polymorphism in Pharmaceuticals

Polymorphs are unexpected Polymorphs seems to be more common for compounds with :
Low solubility in water Organic salts Formation of hydrates - for larger molecules Organic solvates neutral compounds with larger molecular weights. Compounds with molecular weight below 350

Griesser and Burger 1999

Occurrence of Polymorphism in Pharmaceuticals Compilatios of polymorphism in pharmaceuticals

Kuhnert-Brandstatter (1965) Kuhnert-Brandstatter (1971) Borka and Haleblian (1990) Borka (1991-1995) review included in EP (Fas 1-13/13-19) Griesser and Burger (1999)
EP (1997). 559 polymorphic forms, solvates (including hydrates) Merck Index (1997). 10330 compounds in only 140 (1.4%) are
specifically noted as polymorphic, 540 (0.5%) have been specified as solvates.

Current state of awareness of polymorphism on the part of compilers of such compendia and references works
FRECUENCY OF POLYMORPHS IS UNDERSTIMATED

Solid State Properties: Polymorphs

Polymorphism

Solid State Properties are affected :

Mechanical Superficial Spectroscopic Thermodynamic Kinetic Reactivity and/or Stability

Polymorphism of Roy
N O N O H N C

YN MP = 98 oC = 104.1 Y MP = 109.8 oC = 104.7

S CH3
5-methyl-2-[(2-nitrophenyl)amino]-3-thiophenecarbonitrile

R MP = 106.2 oC = 21.7

OP MP = 112.7 oC = 46.1 ON MP = 114.8oC = 52.6

ORP = 39.4

Yu et al J.Am.Chem.Soc. 122,585 (2000)

Solid State Properties: Polymorphs

Polymorphism Pharmaceutical and biopharmaceutical properties are affected :
Density Melting point Higroscopicity Electrical and optical properties Physical and chemical stability (Reactivity) Dissolution velocity Apparent Solubility Drug product stability, dissolution and bioavailability Quality, safety and efficacy of drug product

Bioavalilability
Pharmacological effect

pH = 1 - 3
absorption
hepatic metabolism

dissolution disintegration gastric emptying rate

pH = 5 - 7
intestinal transit rate dissolution
intestinal metabolism

absorption disintegration

faeces

clearance

Bioequivalence
Bioequivalent
10

Concentrao

8 6 4 2 0 0 2 4 6 8 10 12

Polymorph I

Tempo

No Bioequivalent
10

Concentrao

8 6 4 2 0 0 2 4 6 8 10 12

Polymorph II Membrane

Tempo

Bioavailability
Chloranfenicol Palmitate : A, B, C and amorphous. B and amorphous ACTIVE USP controls Pol. A (< 10%)
Form B Form A

Solution Form III Form I Dihydrate

Carbamazepine: four forms, hydrate, solvates and co-crystals USP controls Pol. III (IR - DRX)

Bioavailability
Carbamazepine: four forms, hydrate, solvates y co-crystals USP controls Pol. III (IR y DRX) Bioavailability
Pol. I

Crystal Structure
Pol. II

Solution Form III Form I Dihydrato

Pol. IV Pol. III

Ritonavir (AIDS drug) Conformational Polymorphism Abbot Laboratories

Ritonavir: Polymorphism

IV

mp 122 C

mp 125 C

mp 80 C

mp 97 C

mp 116 C

Launch in 1996 Commercial 1996 Summer of 1998 Summer 1998 TransForm 2002 6 week effort 2002 5 forms identified.

Ritonavir: Polymorphism

Polymoph (I)

Polymoph (II)

RITONAVIR: SOLUBILITY

Ethanol /water
Form I Form II

99/1
90mg/mL 19mg/mL

90/10
234 60

75/25
170 30

Polymorphs = Hurricanes !!

Relevance of Polymorphism: ANVISA (Brazil)

Resolution - RDC n 135 of 29 May 2003 Republished in the D.O.U of 08/12/2003 TECHNICAL REGULATION FOR GENERIC DRUGS Resolution - RDC n 136 of 29 May 2003 D.O.U 06/02/2003 Regulates the registration of New Drugs TECHNICAL REGULATION FOR NEW OR REFERENCE DRUGS WITH SYNTHETIC OR SEMI-SYNTHETIC ACTIVE INGREDIENTS

Relevance of Polymorphism: ANVISA (Brazil)

Resolution - RDC n 135 of 29 May 2003 Republished in the D.O.U of 08/12/2003 TECHNICAL REGULATION FOR GENERIC DRUGS This regulation serves to establish the technical precepts and procedures for the registration of Generic Drugs in Brazil, as follows: I. Definitions used for the registration of Generic Drugs.
II. Pre-registration measures.
III. Documentation for registration. IV. Drug products that will not be accepted as Generic. V. Post-registration measures. VI. Criteria for the prescription and dispensation of Generic Drugs

II Pre-registration measures
12. Quality control report of the raw materials

12.2.8. Information and determination of the probable polymorphs and the analytical methodology for active ingredients that present polymorphism;

Relevance of Polymorphism: ANVISA

Resolution - RDC n 136 of 29 May 2003 D.O.U 06/02/2003 Regulates the registration of New Drugs TECHNICAL REGULATION FOR NEW OR REFERENCE DRUGS WITH SYNTHETIC OR SEMI-SYNTHETIC ACTIVE INGREDIENTS
h) Technical information on the active ingredient(s) as follows, when fitting: h01) structural formula; h02) molecular formula; h03) molecular weight; h04) complete synonyms and references; h05) physical form of the salt; h06) fusion point; h07) solubility; h08) specific optical rotation; h09) organoleptic properties (color, smell, texture ...); h10) possible isomers (structural, geometric, optical ...); h11) polymorphism, discriminating the characteristics of the used polymorph

and of

others related to the active ingredient;

h12) describe the salt/base proportion and any excesses used; h13) molecular infrared structure; h14) other analyses necessary for the correct identification and quantification of the molecule(s) submitted by the manufacturer or as determined by ANVISA.

Relevance of Polymorphism: ANDA (FDA)

Guidance for Industry: Pharmaceutical Solid Polymorphism Chemistry, Manufacturing and Control Information

Food and Drug Administration (FDA) Center for Drug Evaluation and Research (CDER) July 2007

Relevance of Polymorphism: ANDA (FDA)

Chemistry, manufacturing and controls (CMC) information MUST be submitted to support the approval of an ABBREVIATED NEW DRUG APPLICATION (ANDA) This guidance is intended to assist applicants with the submission of ANDAs when a drug substance exists in polymorphic forms Specifically, this guidance provides: - FDA recommendations on assessing sameness when the drug substance exists in polymorphic forms - Decision trees that provide recommendations on monitoring and controlling polymorphs in drug substances and/or drug products

Relevance of Polymorphism: ANDA (FDA)

Characterization of Polymorphs (Guidance for Industry: ANDAs: Pharm. Solid Polymorphism
(Pag. 2-3) )
There are a number of methods that can be used to

characterize polymorphs of a drug substance. Demonstration of a nonequivalent substance by single crystal X-Ray diffraction is currently regarded as the definitive evidence of polymorphism. X-ray powder diffraction can also be used to provide unequivocal proof of polymorphism.
Other methods, including microscopy, thermal analysis and

spectroscopy (IR, Raman, solid state NMR) are helpful to further characterize polymorphic forms

Relevance of Polymorphism: ANDA (FDA)

Relevance of Polymorphism: ANDA (FDA)

Chlorpropamide (CPD)

CHLORPROPAMIDE
Chlorpropamide (CPD) belongs to the sufonyl urea class of oral hypoglycemics United State Pharmacopeia , British Pharmacopeia, European Pharmacopeia Five different polymorphs of CPD have been identified to date (I, II, III, IV, V), of which 3 (A,B,C) are most commonly referred to in the published literature although with very confuse nomenclature

CHLORPROPAMIDE: Solid state characterization

Preparation methods
Crystallization under different conditions (bibliography) Crystallization methods in order to obtain pure polymorphs I(C), II, III(A), IV. (confused bibliography: methods and nomenclature )

Methodology
XRD lab. and High Resolution Powder X-Ray Diffraction Structure Determination using Synchrotron Radiation Scanning Electronic Microscopy (SEM) DSC (lab and SR simultaneously with XRD) solid state Nuclear Magnetic Resonance and Nuclear Quadrupolar Resonance Spectroscopy Intrisic Dissolution Velocity (IDV) (Correlation to biofarmaceutical properties)

Classification and correlation of polymorph CPD

Bibliography and this work (Nomenclature)
Simmons (1973) Burger (1975) Koo (1980) Al-Saieq (1982) Tudor (1993) Otsuka De Drebushchak (1995) Villiers (2006) (1999) Our Work (2006-2007)

Polymorph A Mod. I

Form I

--

Form C Form C

Polymorph I (C)
PDXRD

Polymorph B Mod. II

Form II = V Polymorph B

--

Form B Polymorph Beta (B)

Polymorph II

Polymorph C Mod. III ---Mod. IV Mod. V Solvate Benzene

A (alpha)

Form IV Form III ---

Polymorph A Form A Form A ----------

Polymorph III (A) Polymorph IV

Single crystal XRD

---

Preparation methods: Crystallization conditions

General crystallization conditions Solvents different polarities Concentration of the solutions (super saturated, saturated, diluted) Cooling speed (quenching, slow) Temperature (room or lower than room temperature)

CHLORPROPAMIDE
Preparation methods: Crystallization conditions

Polymorph I (C)

Polymorph II

Polymorph III(A) Commercial raw material Crystallization in different solvents (ej: ethanol, methanol, acetona, propanol, isopropanol, chloroformo, ethyl acetate, acetonitrilo)

Polymorph IV Saturated solution of hexanol at 60C, quenching cooling Observation: single crystal was obtained

Controlled heating Fused raw material at rate up to 110C of Pol. 130C for 2 hours and III sample (raw controlled cooling rate up to material) room temperature Observation : single crystal was impossible to obtain !!!! (only powder sample)

LNLS Laboratorio Nacional de Luz Sincrotron: Powder X-Ray Diffraction with Synchrotron Light

Beamline D11 A - SAS

Beamline D10B - XPD

LNLS Laboratorio Nacional de Luz Sincrotron

Beamline D10B XPD X-Ray Powder Diffraction

Synchrotron light

Detector Goniometer

Sample

SEM

CHLORPROPAMIDE
SR-PXRD

CPD Pol. I: Crystal Structure

Synchrotron Radiation X-Ray Powder Diffraction Crystal Structure Determination (poster 44)

CPD Pol. I: Crystal Structure

Molecular conformation
Polymorph I Ortorrhombic Polymorph III Ortorrhombic

Conformational Polymorphism

Polymorphs CPD : Molecular Conformation

Polymorph I (yellow) Polymorph II (red) Polymorph III (cyan) Polymorph IV (magenta)

Polymorphs CPD: Molecular Packing

Pol.I Pol.III

Pol.II

Pol.IV

Thermal Analysis CPD: conventional

DSC analysis: DSC MDSC 2920, TA Instruments

-0.5

Polimorfo I (C) Polimorfo II Polimorfo III (A)

polimA clorPD107.000 clorPD105 clorPD103

-2.5

Heat Flow (W/g)

-4.5

-6.5

-8.5

-10.5 25
Exo Up

Polimorfo IV
35 45 55 65 75 85 95 105 115 125 135

Temperature (C)

Universal V2.5H TA Instruments

Spectroscopies CPD: NMR solid state

C= O

P o lim o rfo I

P o lim o rfo II

P o lim o rfo III

P o lim o rfo IV

150

100

50

[p p m ]

Thermal Analysis CPD: conventional

0

Rate Heating: 5C/min

Pol I (C)

melting 130C
0.0

-2

Heat Flow (W/g)

Pol III (A)

-4

recrystallization 126C

-0.2

Pol II

-0.4

95C
Pol IV

Heat Flow (W/g)

melting 124C
-6

118C

-0.6

-0.8

melting 130C (pol I)

-8 20
Exo Up

104C

-1.0
40 60 80 100 120 140
Universal V2.5H TA Instru

Temperature (C)

-1.2

123C
-1.4 20
Exo Up

40

60

80

100

120

140

Temperature (C)

Universal V2.5H TA Instru

DSC MDSC 2920, TA Instruments.

Thermal Analysis CPD: Synchrotron Radiation

LNLS Beamline D11 A SAS (DSC and DRX simultaneously) DSC Cell (THM 600, LINKAM)

Detector DSC cell

sample

Thermal Analysis CPD: Synchrotron Radiation

Pol III: calentamiento a 5C/min Polimorfo III (A) 5C/min
-500
III

XRD
Pol III
DSC counts

-1000

Pol I

III+I I I

-1500
127 C

-2000

-2500
134 C fusin I

-3000 40 60 80 100 120 140

Temp (C)
12 seg / patrn DRX

Thermal Analysis CPD: Synchrotron Radiation

Pol II: calentamiento a 5C/min

XRD
Pol II
DSC counts

-500

Pol I

-1000

II

-1500

II I I

-2000

105 C 117 C

-2500
125 C fusin I

-3000 40 60 80 100 120 140

Temp (C)
12 seg / patrn DRX

Thermal Analysis CPD: Synchrotron Radiation

Pol IV: calentamiento a 5C/min
-500

XRD
DSC counts

-1000

IV

Pol I -1500
IV III IV III I I 110 C

Pol III

-2000

-2500

-3000
123 C fusin I

40

60

80

100

120

140

Temp (C)
12 seg / XRD pattern

Pol IV

Intrinsic Dissolution Velocity (IDV) CPD

Dissolution equipment and IDV accessory (United State Pharmacopeia)

Polymorphs CPD: Molecular Packing

Pol.I Pol.III

Pol.II

Pol.IV

Intrinsic Dissolution Velocity (IDV) CPD

IDV analysis
0.016 0.014 0.012
Aprom Bprom conc05 conc104

pol II (B)

Polymor
pol III (A) pol I pol IV

Rate

IDV (mg-min-cm2) 0.0349 0.0522 0.0372 0.0362

Conc (mg/ml)

0.010 0.008 0.006 0.004 0.002 0.000 0 20 40 60 80 100 120

I (C) II (B) III (A) IV

7.75 e-5 11.60 e-5 8.27 e-5 8.05 e-5

Tiempo (min)

Conditions: Medio: 225 ml H2O 37C, 150 rpm

IDV = Rate(mg/ml/min) * Volumen total (225 ml) Area (0.5 cm2)

Progesterone
(poster 45)

Progesterone: Polymorphs
Form Alfa (MP 129 C pol. More stable less soluble )
XRD SEM

Form Beta (MP 121 C more soluble )

XRD SEM

Progesterone in different matrix

Form alfa

XRD
Polyisoprene
d) D

Form Beta

Silicone
C c) B b)

Form alfa

Silicone vulcanized

Form alfa

raw mat.
2-Theta - Scale

a) A

Form alfa

5 66 77 88 99 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 5 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 2-Theta-scale

Progesterone: Polymorphs
Forma beta
Progesterona liberada [ug/cm2]
1000 800 600 400 200 0 0 200 400 600
Silicona Polyisoprene (PI)

Forma alfa

Tiempo [horas]

Progesterone release during 10 days matrices loaded with 20 mg/cm2. - Silicona () - Polyisoprene ()

Progesterone: Polymorphs
Average blood plasma profiles obtained following insertion of -CIDR-Pfizer Argentina (marketed) (1.900g) (back)
0 50 100 150 200

Plasma progesterone [ng/ml]

7 6 5 4 3 2 1 0

T ime [hours]
c ont rol 250 mg 500 mg 750 mg

Innovation of Ceprocor transfered to Veterinary Company

-PI intravaginal devices with 0.250g (blue), 0.500g (green) 0.750g (red) progesterone initial loadings. (n=3)

Patents Efavirenz

Polymorph Patents: Efavirenz (AIDS drugs)

Molcula DRX (forma I anhidra)

10

11

12

13

14

15

16

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19

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23

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25

26

27

28

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31

32

33

34

35

36

37

38

39

40

2-Theta - Scale

SEM

Polymorph Patents: Efavirenz

Polimorfos Referencia US 5,965,729 (12-101999)
Form I

Referencia WO 99/64405 (16-12-1999)

Referencia WO 2006/018853 (23-02-2006)

Referencia WO 2006/030299 (23-03-2006)

Referencia WO 2006/040643 (20-04-2006)

Referencia WO 2006/040643 (20-04-2006)

Forma I

Form 1 (algunos picos de menos) Form 4 (pico 3.62tetha diferente) Form 3 (algunos picos de menos) Form 2 (diferente a otras) Form 5 Form H1 Forma amorfa Form II

Form 1 (1 pico mas en aprox 102tetha)

Form (similar a 1, menos similar a I)

Forma II

Form II

Forma III

Form III

Forma 2

Forma 5 Forma H1 Forma Amorfa

Form Forma Amorfa Form Form Form Form N, O, P (bastante similares) Form 2 (casi amorfo)

Forma Alfa Forma Beta Forma Delta Forma N (O,P)

Polymorph Patents: Efavirenz

Polymorph I Polymorph Beta

10

11

12

13

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19

20

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40

2-Theta - Scale

Polymorph Patents: Efavirenz

XRD Pattern (black): single crystal data (New polymorph?)
Pol I Pol II

10

11

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2-Theta - Scale

2-Theta - Scale

Pol III

Pol 5

10

11

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2-Theta - Scale

2-Theta - Scale

Solange Wardell and S.L. Cuffini

Mebendazole

Mebendazole: Polymorphs
Efficacy of single-dose mebendazole, polymorphic forms A and C in the treatment of hookworm and Trichuris infectionsP. Charoenlarp, J. Waikagul, C. Muenoo, S. Srinophakun and S. Kitayapo. S. Asian J. Trop. Med. Public Health (1993); 24(4):712-716.
Polymorph A INACTIVE 2 10 20

Polymorph C

ACTIVE

Polymorph B

ACTIVE

2-Theta - Scale

30

40

In more than 900 children, Pol. A presented the some results than placebo

Mebendazole: Polymorphism in International Pharmacopeias USP (30)

No indication of

EP 2000
Indication of

Polymorphism presence Techniques in solids: IR ( WITH recrystallization) Standard Ref: MBZ Pol. B No specification of Polymorph Dissolution Test no specific for detection polymorph A or C

Polymorphism presence (> 500 APIs) Techniques in solids: IR (WITHOUT recrystallization) Standard Ref.: MBZ Pol. C

Mebendazole: Polymorphs

Pol. C

Pol. A

Our Pharmacopeias Brazil and Argentina ??

Mebendazole: Suspensions
Quantitaive Methods by X-Ray Diffraction

Pol. C to Pol. A 90 days

Time / days

Facilities de LRA-Sector XRD

D8 Advance BRUKER Standards NIST ( XRD) Data bases PDF CDB LRA (own DB) aprox. 200 APIs

Laboratory Certifications since 2004 - ISO 9001:2000 2005 - IRAM 3800:1998 2005 - OHSAS 18001:1999

Unidad Ceprocor-ACC
Responsible LRA-Sector DRX: Dra. Silvia Cuffini Sonia Faudone, Maribel Ferro y Eugenia Sordello Responsible del LRA-LAF: Dra. Viviana Dabbene Eugenia Quinzio, Gabriela Castelli, Cristian Casado, Cecilia Olmos, Silvia Farfn, Carolina Rizzi Responsible del LBT I: Dr. Ismael Bianco
Dr. Dante Beltramo, Valeria Herrera

Collaborations
Fac. Ciencias Qumicas. Universidad Nacional de Crdoba (ARG) Fac. Matemtica, Astronoma y Fsica. Universidad Nacional de Crdoba (ARG) Dpto. de Fsica. CNEA (ARG) Fac. Qumica, Bioqumica y Farmacia. Universidad Nacional de San Luis (ARG) IF Sao Carlos. USP (BR) FCF Riberao Preto. USP (BR) Hospital das Clinicas. Fac. de Medicina. USP (Riberao Preto-BR) Fac. de Ciencias Farmaceuticas. UNESP (Araracuara-BR) Dpto. de Fsica. UFC (Fortaleza BR) Far-Manguinhos - FIOCRUZ (Rio de Janeiro BR) Laboratorio Nacional de Luz Sincrotron (LNLS) e LME / LNLS (Campinas-BR) Dpto. De Fsica. Universidad de Chile (Santiago de Chile. Chile) Frankfurt University. (Frankfurt - Germany)

For Pharmaceutical Industry

Polymorphs = Hurricanes !! We know that they exist, although it is not know when they will be appear and what kind of damage they will cause

For me Polymorphs = Challenges !! To Know it to control it

Enjoy!!!
Polymorphs, LAPOLC and FORTALEZA!!!

Muito Obrigado!!! scuffini @ ceprocor.uncor.edu scuffini @ yahoo.com