In Situ Metastable Form: A Route for the Generation of Hydrate and

Subscriber access provided by UNIVERSITY OF ADELAIDE LIBRARIES

Article

In-Situ Metastable Form : A Route For The Generation Of Hydrate And Anhydrous Forms Of Ceritinib Ramanaiah Chennuru, Ravi Teja Koya, Pavan Kommavarapu, Saladi Venkata Narasayya, Prakash Muthudoss, R. Ravi Chandra Babu, and Sudarshan Mahapatra Cryst. Growth Des., Just Accepted Manuscript • DOI: 10.1021/acs.cgd.7b01027 • Publication Date (Web): 11 Oct 2017 Downloaded from http://pubs.acs.org on October 12, 2017

Just Accepted “Just Accepted” manuscripts have been peer-reviewed and accepted for publication. They are posted online prior to technical editing, formatting for publication and author proofing. The American Chemical Society provides “Just Accepted” as a free service to the research community to expedite the dissemination of scientific material as soon as possible after acceptance. “Just Accepted” manuscripts appear in full in PDF format accompanied by an HTML abstract. “Just Accepted” manuscripts have been fully peer reviewed, but should not be considered the official version of record. They are accessible to all readers and citable by the Digital Object Identifier (DOI®). “Just Accepted” is an optional service offered to authors. Therefore, the “Just Accepted” Web site may not include all articles that will be published in the journal. After a manuscript is technically edited and formatted, it will be removed from the “Just Accepted” Web site and published as an ASAP article. Note that technical editing may introduce minor changes to the manuscript text and/or graphics which could affect content, and all legal disclaimers and ethical guidelines that apply to the journal pertain. ACS cannot be held responsible for errors or consequences arising from the use of information contained in these “Just Accepted” manuscripts.

Crystal Growth & Design is published by the American Chemical Society. 1155 Sixteenth Street N.W., Washington, DC 20036 Published by American Chemical Society. Copyright © American Chemical Society. However, no copyright claim is made to original U.S. Government works, or works produced by employees of any Commonwealth realm Crown government in the course of their duties.

Page 1 of 33

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60

Crystal Growth & Design

In-Situ Metastable Form: A Route For The Generation Of Hydrate And Anhydrous Form Of Ceritinib Ramanaiah Chennuru#$, Ravi Teja Koya#, Pavan Kommavarapu#, Saladi Venkata Narasayya#,Prakash Muthudoss#, R. Ravi Chandra Babu$*and Sudarshan Mahapatra#*

#-Centre for Excellence in Polymorphism and Particle Engineering, Integrated Product Development (IPDO), Dr. Reddy's Laboratories Ltd., Bachupally, Hyderabad, Telangana, India $-Department of Chemistry, College of Science, GITAM University, Visakhapatnam, Andhra Pradesh, India

_________________________________________________________________________

*Corresponding author Dr. Sudarshan Mahapatra

Center for Excellence in Polymorphism and Particle Engineering, Integrated Product Development (IPDO), Dr. Reddy's Laboratories Ltd., Bachupally, Hyderabad, Telangana State -500090, India Email: [email protected] Contact No: +91-40- 44346040 Fax No: +91-40-4434 6164

ACS Paragon Plus Environment


1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60

Abstract Ceritinib is an anaplastic lymphoma kinase (ALK) inhibitor used for the treatment of ALKpositive metastatic a non-small cell lung cancer (NSCLC). This BCS class IV drug is developed by Novartis and traded under the name Zykadia®. As on date two forms [Form-A (marketed form) and B] of ceritinib are disclosed in international patent application US 2013/0274279 A1. However, crystal structure, insight on any solid form of this compound is not available in the literature. In order to achieve better physico-chemical properties compared to known solid form of this compound, novel polymorph identification is chosen as one of the challenging path to address the issue. In our comprehensive polymorph screening, including in-silico and experimental investigation, we could discover three novel solid forms of ceritinib. Out of these three solid forms two are neat (Form-1 and Form-3) and remaining one is a hydrate (Form-2). All synthesizes forms are further characterized by PXRD, DSC and FT-IR. It is interesting to note that the discovery of this hydrate is in sync with the prediction done using COSMO-RS theory (COSMOTHERM-X software). Current article includes the first single crystal structure of ceritinib Form-1. All Forms (Form-1, 2 and 3) of ceritinib are subjected to physico-chemical property evaluation like solubility in buffers with a pH range of 1 to 7, dissolution and stability. In Aqueous and pH 4.5 (Acetate buffer) the solubility of Form-2 & 3 is high compared to Form1. Whereas in 0.1N HCl and 0.01N HCl Form-1 is having a higher solubility compared to Form2 and 3. Six moth stability study indicates that all forms (Form-1, 2 and 3) are stable in ICH stability conditions like Accelerated (40°C ± 2°C, 75% RH ± 5%RH), Long term (25°C ± 2°C, 60% RH ± 5%RH) and Low temperature (2-8°C) conditions. A thorough polymorph screening protocol, including in-silico prediction, single crystal structure, and physico-chemical properties of different forms and structure property correlation for ceritinib are enlightened in the current paper.

Key words: Single crystal, COSMOTHERM-X, Polymorphism, Solid Form, Ceritinib.


Page 2 of 33

Page 3 of 33

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60


1. Introduction In recent decades both academic groups and pharmaceutical companies have shown considerable interest in the application of crystal engineering principles to control and tune the physico-chemical properties of active pharmaceutical ingredients (API’s).1-2 Further, in literature it is indicated that a majority of drug molecules (>80%) fail at the late stage of drug formulation and pre-clinical studies due to issues related to low solubility and poor bioavailability. 1-2 Salt formation is found to be the most common and well-practiced approach to improve the physicochemical properties like: solubility, stability, dissolution rate, melting point, filterability, etc. 3-7 In this regard co-crystals can also be considered as an equal competitor to salt. However, salts are sometimes more likely to form hydrates and have an inherent tendency towards hygroscopicity compared to co-crystals.8-9 On the same line polymorphism in API’s is considered as an alternate and over decades, it has become a major area of drug research. Polymorphs exhibit unique physicochemical properties, such as melting point, solubility, bioavailability, stability, color, mechanical properties, correlating to their unique crystal structure and packing.10-12 Improved aqueous solubility is a primary driving force in the development of novel solid- form (amorphous, polymorphs, co-crystals, hydrates/solvates, and salts) selection.1317

Polymorphs and hydrates are often obtained by slow solvent evaporation or cooling

crystallization dealing with the saturated solution of the target compound.18-19 Other crystallization approaches involving variation in crystallization temperature,20-21 milling condition,22 solvent polarity,13humidity24 etc., can be explored during polymorph screening and crystallization. However, the discovery of polymorph/hydrate has been always a challenging task. In the current study, we have demonstrated how the pH modulation can act as one of the potential approaches to isolate novel solid form. The challenge of low aqueous solubility offers an ideal situation for the application of crystal engineering, 25 concept to mitigate the issue with solubility, dissolution rate and bioavailability. We have conducted polymorph screening in ceritinib (BCS-IV) drug with an aim to identify novel, stable, scalable polymorph with better/similar physico-chemical properties like Form A. In the current research paper, we have reported two anhydrous (Form-1 and Form-3) and one hydrate (Form-2) form of ceritinib and a comprehensive study of their physico-chemical properties. Ceritinib is an anaplastic lymphoma kinase (ALK) inhibitor used for the treatment of ALK-positive metastatic non-small cell lung cancer (NSCLC). Ceritinib is used for the treatment



1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60

of NSCLC following treatment with Crizotinib. It is developed by Novartis and traded under the name Zykadia®. It was approved by FDA on April 7, 2014. Ceritinib is formulated as hard gelatin capsules with 150 mg strength with a maximum daily dose of 750 mg. So far two polymorphs (Form-A and B) of ceritinib are reported in literature. In our polymorph screening, we have isolated three crystalline forms of ceritinib and named as Form-1, 2 and 3. The powder X-ray diffraction pattern (PXRD) and DSC thermogram of Form-1 (Neat form) matches with Form-A, disclosed in international patent application US 2013/0274279 A1. Form-2and Form-3 is found to be hydrate and neat form respectively. In-silico technique is always having a less importance in generic drug development. However, in the current paper we have discussed how it can be utilized effectively to match the pace of solid form screening (polymorph or hydrate) at the earliest. Single crystal structure analysis of marketing solid form (Form-1) and characterization of synthesized solid forms are reported in the paper. Comprehensive physicochemical properties like solubility, powder dissolution performance assessment and stress/hygroscopic stability studies for all forms of ceritinib are explained thoroughly in the current paper.

2. Experimental Section 2.1 Materials and Methods Ceritinib API was provided by Dr Reddy’s Laboratories Ltd., and it is further purified by recrystallization from acetone. All other chemicals were of analytical or chromatographic grade. Water purified from a deionizer-cum-mixed-bed purification system (Millipore®, USA) was used in the experiments. 2.2Preparation of Ceritinib anhydrous and hydrate forms 2.2.1 Ceritinib Form-1 Ceritinib input material was dissolved in acetone & the solution was heated up to 50°C followed by cooling down to room temperature (~25°C) lead to precipitation. The precipitate was filtered and suck dried under vacuum. The formation of Form-1 was confirmed by PXRD, DSC, TGA and MC (See supporting information figure-S1, S4, S7 and S10). For single crystal analysis,


Page 4 of 33

Page 5 of 33

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60


colorless block-shaped single crystals were obtained by vapor diffusion method in 3-4 days using acetone as solvent and n-heptane as anti-solvent. 2.2.2 Ceritinib Form-2 Ceritinib (3.0g) was taken in a 100ml Easymax® reactor and 15ml acetic acid was added to it at room temperature. The pH of the resulting solution was found to be around 2.5. The clear solution was maintained at room temperature over constant stirring using an overhead stirrer with 350 rotations per minute (RPM). The pH of the reaction was increased to 11-12 by using 1M NaOH solution, followed by maintenance of reaction mass at room temperature for 24 hours leading to precipitation. The precipitate was filtered and suck dried under vacuum at room temperature. The isolated material was dried in the Air Tray Dryer (ATD) at 40°C for 2 Hrs. The formation of a new solid hydrated form was confirmed by PXRD, DSC, TGA and KarlFischer titration for moisture content (MC) (See supporting information figure-S2, S5, S8 and S11). This hydrate is named as Form-2. The attempts to grow single crystals of Form-2 by slow solvent evaporation, vapor diffusion and cooling-crystallization, resulted Form-1 (Form-2 converts to Form-1 in above mentioned conditions). So, single crystal structure has not been solved for Form-2. However, the phase purity is confirmed by indexing the PXRD pattern and described in the following section. 2.2.3 Ceritinib Form-3 Form-2 (3.0 gm.) was taken in an Air Tray Dryer (ATD) and maintained at 120°C for 10 minutes to generate Form-3. The obtained material is characterized by PXRD, DSC, TGA and MC and confirmed to be a neat solid form (See supporting information figure-S3, S6, S9 and S12). 2.3 Instrumentation 2.3.1 Differential Scanning Calorimetry (DSC) DSC thermograms of all samples were recorded on a Thermal Advantage (TA) Discovery instrument by heating the samples at a heating rate of 10° C/min up to 200° C with the continuous purging of dry nitrogen gas at a flow rate of 50 mL/min.



1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60

2.3.2 Thermogravimetry analysis (TGA) TGA thermograms of all samples were recorded on a Thermal Advantage (TA) Q 500 series machine by heating the samples at a rate of 10° C/min up to 200° C with the purging of dry nitrogen gas with a flow rate of 40 mL/min to the balance and 60 mL/min to the furnace. 2.3.3 Powder X-ray Diffraction (PXRD) All X-ray powder diffraction data were collected on a PANalytical X’Pert PRO diffractometer (X’cellerator detector, Cu-anode, 45KV, 40mA, Brag-Brentano geometry) using the 2θ scan range, step size, and exposure time of 3-40°, 0.03° and 1200 s/step respectively. 2.3.4 Single-Crystal X-ray Diffraction Single crystal data sets for ceritinib Form-1 were collected in open mounting condition on an Oxford-Super Nova diffractometer with a CCD detector. The crystal structure was solved using direct methods on a reasonably good quality data set obtained from a carefully chosen crystal. 2.3.5 Purity by HPLC All samples purity was analyzed for the purity by using HPLC. The Purity of the samples is calculated by % area normalization. Samples were analyzed using the following parameters and chromatographic conditions, column: YMC Pack ODS-A (150 mm x 4.6 mm, 3.0 µm); flow rate: 1.0mL /min; Injection volume: 10.0µL; Column oven temperature: 30°C; Run time: 60 minutes; Detector wavelength: 215 nm. Gradient program-Time (Minutes) 0, 43, 53, 54 and 60 with % Mobile phase-A 90, 10, 10, 90 and 90 respectively. Sample Preparation: Ceritinib sample (25 mg) was weighed and transferred into a 50 mL volumetric flak followed by the addition of 40 mL of diluent (Acetonitrile: Water 50:50% v/v) and sonication to dissolve the material. The volume was made up to mark with diluent and mixed well. The samples were filtered through 0.22 µm PVDF (Polyvinylidene fluoride) syringe filter and the solution was collected in HPLC vials before analysis. Procedure: Blank (Acetonitrile: Water 50:50% v/v) was injected before to test solution into the chromatography system. The purity of the sample was calculated by percentage area normalization.


Page 6 of 33

Page 7 of 33

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60


2.3.6 Water Content by KF apparatus All samples were analysed for water content by using Karl-Fischer titration method (Metrohm). 100.00 mg of sample was weighed into the titration vessel and titrated using Karl-Fisher reagent to calculate the percentage of water content. 2.3.7 Solubility determination Equilibrium solubility of Form-1 and the prepared novel polymorphs (Form-2 and Form-3) were determined in Ultra-pure water (Millipore®, USA) and multi-media buffers. The selected buffers in this regard are 0.1N HCl, 0.01N HCl, pH 4.5 Acetate buffer and pH 6.8 Phosphate buffer. Excess amount of sample was added in to selected media(water and buffers)and the samples were shaken for 24 h at 37°C in a horizontal orbital shaker (n =3) at 200 RPM speed. After the requisite time the supernatant was filtered through 0.45 mmµ PVDF syringe filter and the filtrate was assayed Spectro photometrically by HPLC at 215 nm using YMC Pack ODS-A, 150 mm x 4.6 mm, 3.0 µm column. 2.3.8 The drug release rate studies The drug release profile was studied using USP apparatus II (paddle) method using Lab India dissolution apparatus. Dissolution studies were carried out in 900 mL 0.01N HCl (pH 2.0) at 37° ± 0.5° C with a stirring of 60 RPM. An amount equivalent to 750 mg of ceritinib samples were added to the dissolution medium. 5.0 mL of sample was withdrawn after 5, 10, 15, 20, 30, 45 and 60 minutes and equivalent volume is replaced by10 mL fresh 0.01 N HCl. The solutions were filtered through 0.22 mm membrane (µ PVDF syringe)filter and the concentration of ceritinib was determined spectrophotometrically at 215 nm using YMC Pack ODS-A, 150 mm x 4.6 mm, 3.0 µm column. 2.3.9 pH study pH study was conducted for all solubility samples kept for 24 hour time point in different buffers (0.1N HCl, 0.01N HCl, pH 4.5 Acetate buffer and pH 6.8 Phosphate buffer). After 24 hours incubation at 200 RPM at 37°C in different buffer the samples were withdrawn and pH of initial and final were measured using Metrohm pH meter.



1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60

Page 8 of 33

3. Results and discussion Literature search (CSD, version-5.36, November 2014) on ceritinib resulted no hits. As of today, there is no crystal structure available in literature for ceritinib. 3.1.COSMOtherm COSMO-RS (Conductor-like Screening Model for Real Solvents) is a universal theory to predict the thermodynamic equilibrium properties of liquids, which were originally developed by AndreasKlamt.26-29 Propensity to form solvate/co-crystal can be evaluated by calculating the excess enthalpies (Hex). Use of excess enthalpy from COSMO-RS model is very well known in literature.30 On the same line COSMO-RS was used for Ceritinib molecule with the aim to short list the probable solvate/hydrate based on their Hex value. Higher negative excess enthalpies (Hex) for solvent and active pharmaceutical ingredients (API) indicate higher probability of formation of corresponding solvate. It is noteworthy that the excess enthalpy (Hex) for formic acid, acetic acid and water found to be -3.49, -2.470 and -0.8 respectively. Ceritinib is a strong base so considering the pKa difference it is obvious to expect salt formation with formic and acetic acid. However, the negative excess enthalpy of water indicates a higher probability of hydrate formation. Hydrates are pharmaceutically accepted; in this regard screening experiments were conducted on ceritinib with a focus to obtain hydrates. The list of probable solvate/hydrate formation with their corresponding Hex values was tabulated in Table-1 and Table-S1 (See supporting information).

Table-1: Excess enthalpy calculation for Ceritinib with different solvents. API

Ceritinib

Solvent

Hex(kcal/mole)

Formic acid

-3.4929

Acetic acid

-2.4763

Water

-0.844

n-Butylacetate

+0.0078

Ethylformate

+0.004159

Anisole

+0.16481

Iso-propylbenzene

+0.21824

Pentane

+0.3914

n-Heptane

+0.59


Page 9 of 33

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60


3.2. Hydrate screening Ceritinib molecular structure indicates presence of eight hydrogen bond acceptors and three hydrogen bond donors. This clearly gives an intuition of strong hydrogen bond with water. It is noteworthy that till to date ceritinib hydrates are not reported in the literature. In this regard hydrate screening for ceritinib was initiated with a background support of COSMORS prediction. There are number of methods reported31for generation of hydrate forms of API which includes (i) Exposure to different RH32 (ii) Slurry in water32at different temperature(III) Temperature cycling of aqueous suspension33 (iv) Slurry in Mixed-solvent33at different temperature(v) Vapor diffusion (vi) Solvent/anti-solvent mixing at different temperature (vii) Heating/Cooling crystallization (viii) Solvent exchange at different temperature (ix) Wet−drying (x) Wet grinding31at different temperature and so on. Most of these crystallization techniques were explored for ceritinib hydrate but each time they ended up with the known anhydrous solid form i.e. Form-1. The experimental trials and their results are tabulated in Table-S2. 3.3. Discovery of Ceritinib novel hydrate via pH modulation crystallization technique Ceritinib has two strong basic cites i.e. piperidine N1 with Pka -10.07 and pyrimidine N4 with Pka-3.059 (Scheme-1). Ceritinib micro species distribution Vs. pH is calculated using Marvin 5.10.1, 2012, ChemAxon.34 This distribution shows that Piperidine N1 can be easily protonated at any pH 8.6 (Figure-1). This pH dependent modulation technique for salt making and breaking was selectively used for the discovery of hydrates of the ceritinib free base. Ceritinib (3.0 gm.) was charged into a RBF and acetic acid was added slowly to the reactor with constant stirring until all material got dissolved. The pH of the resulting solution was measured to be pH= 2.5. Then excess aqueous sodium hydroxide (1M NaOH) solution was added into the RBF until a pH of 11.8 is reached. The reaction mass was maintained for 24 hours with constant stirring. The reaction mass was filtered and formation of hydrate was confirmed by PXRD, DSC and Moisture analysis (KF titration technique). The discovered hydrated solid form is named as Form-2 (See supporting information figure-S2, S5, S8 and S11). Hydrate formation via pH modulation technique is rarely explored in the literature and we believe this could be one of the additional ways to explore hydrates of active pharmaceutical ingredients.



1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60

Scheme-1


Page 10 of 33

Page 11 of 33

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60


Figure-1 Microspecious distribution (%) vs. pH



1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60

Page 12 of 33

3.4. Characterization of Ceritinib Forms: 3.4.1. Crystal structure Analysis Block morphology crystals of Ceritinib Form-1 were obtained from a saturated solution of acetone at room temperature. The crystal structure was solved and refined in a monoclinic cell with space group P21/n. The overlay of experimental and simulated PXRD pattern is shown in Figure 2.The asymmetric unit contains only one molecule of ceritinib in the unit cell. The threedimensional crystal packing in ceritinib Form-1 is supported by intermolecular N5-H5…O3 (2.8395 Å; 128°) hydrogen bonding with a graph set35of S (6). Ceritinib forms an infinite chain along the b-axis via C17-H17…N1(3.4315 Å ; 160°) intermolecular interaction with a graph set35of C (3) (Figure 3-i). In the three-dimensional packing ceritinib molecules form a ring motif via C26H26…N4(3.3208 Å; 139°) hydrogen bonding for a graph set35R44 (20) (Figure 3-ii). Crystal structure

of Form-1 indicates the presence of three types of graph set leading to ring motif and three dimensional chains along b-axis (Figure 3-iii).

Figure-2 Experimental powder X-Ray Diffraction (PXRD) pattern Vs. Simulated PXRD pattern of Form-1.


Page 13 of 33

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60


Figure-3 Three-dimensional packing diagram of CeritinibForm-1 showing (i) Three dimensional chains along the b-axis via C-H…N intermolecular interactions (ii) Ring motif for a graph set of R44 (20) (iii) 3D packing diagram along the b-axis.



1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60

Page 14 of 33

Table-2 Cystallographic details of Ceritinib Form-1.

Parameters

SXRD Refinement Ceritinib Form-1

CCDC Number

1563222

NA

NA

Chemical formula moiety in asymmetric unit.

C28H36ClN5O3S

(C28H36ClN5O3S)●(0.5H2O)

C28H36ClN5O3S

Chemical formula

C28H36ClN5O3S

(C28H36ClN5O3S)●(0.5H2O)

C28H36ClN5O3S

Formula weight

558.13

567.13

558.13

Crystal system

Monoclinic

Triclinic

Triclinic

Space group

P21/n

P1

P1

T [K]

293K

293K

293K

a [Å]

13.302(5)

19.0875(3)

18.8831(7)

b [Å]

8.782(5)

9.8051(7)

15.7279(3)

c [Å]

24.896(5)

9.0839(1)

12.4435(6)

α [°]

90

80.7260(8)

115.2045(1)

β [°]

96.785(5)

104.1594(8)

79.2349(6)

γ [°]

90

118.9312(1)

120.5435(2)

Z

4

2

4

V[Å3]

2888(2)

1777.662(1)

2894.101(4)

Dcalc [g cm–3]

1.284

–1

PXRD Refinement Ceritinib Form-2 Ceritinib Form-3

Μ [mm ]

0.242

-

-

Total reflns.

14058

-

-

Unique reflns.

5874

-

-

Observed reflns.

3913

-

-

R1 [I>2(I)]

0.058

0.0554

0.0485

wR2 (all)

0.094

0.0402

0.0926

Goodness-of-fit

1.018

-

-

Instrument

Bruker Apex Diffractometer

Bruker D8-Advance

Bruker D8-Advance


Page 15 of 33

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60


Table-3 Hydrogen bond metrics in the crystal structure of Ceritinib Form-1. Interaction

D-H/Å

D-A/Å

H-A/Å D-H…A [°]

Symmetry

Ceritinib Form-1 C17-H17…N1

0.93

3.4315

2.54

160

1/2+x,1/2-y,-1/2+z

C26-H26…N4

0.93

3.3208

2.51

139

1-x,-y,-z

N5-H5…O3

0.86

2.8395

2.23

128

x,y,z

N2-H2…O1

0.86

2.6505

2.37

100

x,y,z

N5-H5…Cl1

0.86

2.9901

2.55

113

x,y,z

3.4.2. Powder indexing Powder indexing is used to determine the phase purity of crystalline solid materials.36 In order to determine the phase purity of Ceritinib Form-2 and Form-3; powder indexing was performed with the help of Material studio (Version-8.0). X-ray powder diffraction pattern indexing was carried out using X-cell37/Dicvol38 programs with the option of automatic space group determination in Material Studio. An empty unit cell was created for the cell having best relative figure of merit (FOM) and weighted profile factor Rwp. The unit cell was further refined by Pawley method.39 Powder indexing results of Form-2 and 3 proved them to be unique phase. Powder refinement data of Form-2 and 3 are shown in Figure-4.Crystallographic information of form-2 and 3 are tabulated in Table-2.

Figure-4: Pawley profile fitting of X-ray powder diffraction pattern of Form-2.



1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60

Page 16 of 33

Figure-5 Pawley profile fitting of X-ray powder diffraction pattern of Form-3. 3.4.3. Powder X-Ray diffraction (PXRD) technique Powder X-ray diffraction technique (PXRD) is a vital and predominant tool for the study of polycrystalline materials, and is eminently suited for the routine characterization of polymorphs.40 Polymorphism in Form-1 and 3 is clearly evident by considering their X-ray powder diffraction pattern (Figure-5). In sink to this there are three distinct peaks at 2-θ = 5.42⁰, 9.37⁰ and 15.04⁰in X-ray powder diffraction of Form-3 found to be absent in Form-1. At the same time two peaks at 2-θ=12.78⁰ and 15.60⁰ of Form-1 are absent in Form-3 (Figure-6). Comparing the experimental PXRD patterns of Form-1 and 2, four distinct peaks at 2-θ = 5.05⁰, 9.61⁰, 10.09° and 15.11⁰for Form-2 are absent in Form-1. At the same time three peaks of Form1 at 2-θ 10.63⁰, 13.25⁰ and 17.58° are absent in Form-2 (Figure-5). Comparing the experimental PXRD patterns of Form-2 and Form-3, four distinct peaks of Form-2 at 2-θ 5.05⁰, 9.61⁰, 10.09° and 17.11⁰ found to be absent in Form-3. At the same time three peaks of Form-3 at 2-θ 5.43⁰, 9.36⁰ and 16.40⁰ are absent in Form-2 (Figure-6).


1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60


Intensity in arbitrary unit (a.u.)

Page 17 of 33

Form-1

Form-2 Form-3 10

20

30

Position-(2Ɵ°)

Figure-6 Experimental powder X-Ray Diffraction (PXRD) pattern overlay of Ceritinib Form-1, 2 and 3. 3.4.4. Thermal Technique DSC has distinct advantages over other analytical techniques, including the ease, simplicity and rapidity of the measurements both in terms of identification and characterization. It also provides information on thermodynamic parameters associated with the polymorphic transition.4142

Polymorphism between different solid forms is well established in literature by their

characteristic melting point and enthalpy of fusion.43Ceritinib Form-1, 2 and 3 were subjected to DSC thermal analysis. DSC thermogram of Form-1 shows only a melting endotherm, with Tonset/ Tpeak at 174.30/176.70° C. On the other hand ceritinib Form-2 shows three kinds of events before the melting endotherm. The first broad endotherm is spreading over 40-80°C representing the dehydration of Form-2 and the second endotherm of ∆Htrans=-9.2 J/gwith Tonset/ Tpeak at 99.5/101.9° C represent a phase transformation of Form-2 to Form-3. The third endotherm can be assigned to melting of Form-3 with Tonset/ Tpeak at 157.4/160.5and the immediate exotherm can be related to the re-crystallization of the melt to Form-1 with Tonset/ Tpeak at 161.8/162.8° C (Figure-7). The DSC thermogram of Form-3 shows two independent endothermic events & one exothermic event. The first endotherm and immediate exotherm obtained for Form 3 are related to the melting of Form-3 followed by re-crystallization of Form-1 from the melt. The Tonset/ Tpeak



1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60

Page 18 of 33

obtained for these two events are matching very closely to that obtained for corresponding events in Form-2.The last endotherm of both form Form-2 and 3 overlap with the melting endotherm of Form-1. DSC thermal events for various solid forms of Ceritinib are tabulated in Table-5.

Figure-7 DSC thermogram overlay of ceritinib Form-1, 2 and 3.


Page 19 of 33

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60


Table-5 Melting point temperature and enthalpy values of ceritinib forms. Event

Tonset/Peak(°C) ∆Hf(J g-1)

Endo

174.30/176.70

-158.3

Endo

47/74

---

---

Endo

99.5/101.9

-9.2

--

Endo

157.4/160.5

-15.4

--

Exo

161.8/162.8

+23.1

--

Endo

174.10/176.40

--

--

Form 1-Melting

Endo

157.8/160.9

---

--

------

Endo

161.7/162.9

---

--

------

Endo

174.36/175.85

---

---

------

Form-1 Form 1Melting

Form-2

Form-3

--

--

Dehydration

--

Form-2→3 transition Form-3 melting Melt→Form-1 Recrystallization

----

Form-3 melting Melt→ Form-1 Recrystallization Form 1-Melting

3.4.5. Vibrational Spectroscopy Vibrational spectroscopy in conjunction with other solid state techniques is considered extremely valuable for a compound existing in multiple polymorphic forms.44-46 Infrared spectra of Form 1, Form-2 and Form-3 of ceritinib are shown in Figure -8 and 9and detailed peak assignments are tabulated in Table 1. This molecule contains NH, CH3, O=S=O, C-Cl and aromatic C=C groups, hence a variety of molecular vibrational modes may be expected. It is obvious to expect a vibrational band around 3600 and 3100 cm-1for hydrates. The peaks appearing around 3616 cm1

for Form-2 is attributed the water present in the crystal lattice. Between anhydrates (Form-1 and

Form-3), sharp peaks were observed in the region 3450-3200 cm-1 attributing to NH stretching vibrations. The weak to medium peaks appearing between 3100-3000 cm-1 can be attributed to aromatic H-C=C stretching, 1600-1400 cm-1 attributing to aromatic C=C-C stretching. Moreover, a series of vibrations between 900-650 cm-1 (See Table-6) were found to display pronounced differences between polymorphs which are attributed to the CH out of plane vibrations. This information could potentially point out to the fact that the anhydrous forms are differing in the



stacking as well as H-bonding interactions. Furthermore, the anhydrous Form-3 peak positions resemble Form-2 hydrate that may perhaps indicate Form-3 to have a channel anhydrate structure. Fourier Transform Infrared Spectroscopy along with thermal analysis and XRPD study confirms that ceritinib exist in two anhydrous forms along with a hydrate state. FT-IR vibrations both stretching and bending in Form-1, 2 and 3 are tabulated in table-6.

2.0

Form-3 Anhydrate

1.8 1.6

1.0 0.8 0.6 0.4 0.2

NH Region

1.2

Crystalline Water Peaks

1.4

Absorbance

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60

Page 20 of 33

NH bending and Aromatic C=C Region

Form-2 Hemi-hydrate

0.0 -0.2 -0.4

Form-1 Anhydrate -0.6 -0.8 -1.0

3500

3000

2500

2000

1500

1000

500

Wavenumber

Figure-8 FT-IR spectra of Form-1, 2 and 3.


Form-3 Form-2 Form-1

3418

0.5

Absorbance

(i)

3319 3318

0.4

3310

3440 3361 3440

0.3

3616

(ii)

0.22 0.21

3186


3035 3082 3099

3203

3118

0.20

3051

0.19 0.18

0.2 0.17 3700

3600

3500

3400

3300

3200

3100

3200 3180 3160 3140 3120 3100 3080 3060 3040 3020

Wavenumber


0.40 0.35 0.30

Wavenumber

1571

(iV) 1.0

2798 2730

0.25 0.20

1563

1508

1445

1408

1484

Absorbance

(iii) Absorbance

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60


Absorbance

Page 21 of 33

0.8

1598

1411

1465


1540

0.6 0.4 1476

0.2 0.15 0.0 2950 2900 2850 2800 2750 2700 2650 2600

1650

1600

Wavenumber

1550

1500

1450

1400

Wavenumber

Figure-9 FTIR finger print plots for ceritinib Form-1, 2 and 3 (i) wave number range from 37003100 cm-1,(ii) wave number range from 3200-3020 cm-1,(iii)wave number range from 30002600cm-1,(iv)wave number range from 1700-1400 cm-1.



1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60

Page 22 of 33

Table-6: FT-IR vibrations (stretching/bending) in Form-1, 2 and 3. Band Origin

Wavelength Range

Form-1

H-OH

Form-2

Form-3

3616

N-H

~3450

3440 -

3418 1597 1593 (sh)

3440 3418 1597 1593 (sh) 3118 (vw) 3099 sh 3082 3052 3035

N-H

1650-1550

1598

C=C-H

3130-3070

3118 3051 3035

3118 (vw) 3082

1563

1563 (sh)

1563 (sh)

C=C-C

1615-1580 -

1571

1571

C=C-C

1510-1450

1504

1508

1508

S=O

1350-1300

1313

1314

1313

S=O

1160-1120

1141

1139

1142

Description Channel hydrate, OH Stretch Aromatic 2° Amine Stretch Aromatic 2° Amine Bend

Aromatic C-H Stretch

Aromatic Ring Stretch

Aromatic Ring Stretch Anti-symmetric sulphone stretch Symmetric sulphone stretch

Note: Sh- Shoulder peak,vw-Very weak peak

3.5. Physico-chemical properties evolution 3.5.1. Solubility and powder dissolution Solubility: An important goal of drug development protocol is to improve physico-chemical properties and enhance the drug efficacy. Solubility remains a major challenging factor for BCS class IV drugs (low solubility, low permeability) because of this poor dissolution leading to limited bioavailability.23Ceritinib is a BCS class IV drug having an aqueous solubility of 0.001 mg/ml. We have conducted apparent solubility measurement in water at pH-1.2 (0.1N HCl), pH2.0 (0.01N HCl), pH 4.5 (Acetate buffer) and pH 6.8(Phosphate buffer). It is interesting to note that Form-2 (0.9 mg mL-1) and Form-3 (0.1 mg mL-1) are having a higher aqueous solubility compared to Form-1(0.001 mg mL-1). However ceritinib Form-1 (107.1 mg mL-1) has the


Page 23 of 33

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60


advantage on solubility in acidic pH-1.2 (0.1N HCl) over Form-2 (3.9 mg mL-1) and Form-3 (0.9 mg mL-1). Also, similar solubility trend was observed in pH-2.0 (0.01N HCl) media where Form1 (10.1 mg mL-1) has advantages over Form-2 (4.4 mg mL-1) and Form-3 (1.5 mg mL-1). Further at pH-4.5 in acetate buffer, Form-2 (84.6 mg mL-1), Form-3 (82.1 mg mL-1) have slightly higher solubility as compared with Form-1 (54.1 mg mL-1). As like pH-4.5 in pH-6.8 (phosphate buffer) Form-2 (0.004 mg mL-1) and Form-3 (0.004 mg mL-1) show a slight increase in solubility compared to Form-1(0.02 mg mL-1). Apparent solubility order for different solid form of ceritinib in water at pH-4.5 and 6.8 buffers follows the trend like: Form-2>Form-3>Form-1 and at pH-2.0, Form-1>Form-2>Form-3, after 24 hrs. Solubility of ceritinib Form-1, 2 and 3 are pictorially represented in Figure-10. Solubility and different solid form stability at different pH is tabulated in Table-7.

Figure-10 Solubility profiles of ceritinib Form-1, 2 and 3 (a) After 15 min, (b) After 60 min, (c) After 180 min and (d) After 1440 min.



1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60

Page 24 of 33

Table-7 Solid form stability and apparent solubility at different pH.

Initial Form Form-1

Form After 24 Hrs.

Initial pH

Form-1

Form-2

Form-2

Form-3

Form-3

pH after 24 Hrs.

pH=1.2

pH=2.0

pH=4.5

pH=6.8

Water

1.21

2.01

4.53

6.83

6.87

6.48

6.17

5.62

7.22

6.93

2.00

6.52

6.80

8.33

6.47

1.86

6.73

6.98

8.52

6.90

Powder Dissolution: In vitro powder dissolution studies were conducted for ceritinib Form-1, 2 and 3 -2.0 (0.01N HCl) at 37°C. In order to get uniform particle sizes during the dissolution study, all ceritinib forms (Form-1, 2 and 3) were sieved under 20 size mesh. The dissolution profiles of Form-2 and Form-3 are comparable to Form-1(Figure-8).There is a minor difference in the beginning stage of the release profile of Form-1. In case of Form-2 and Form-3, 90% of the drug gets released in the first five minutes. However, in case of Form-1 it is only 75% and it took 20 minutes to reach a 90% drug release for Form-1. In Vitro powder dissolution profiles of Ceritinib Form-1, 2 and 3 are shown in Figure-11.


Page 25 of 33

120

100

80

% Drug released

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60


60

40

20 Form-1

Form-3

Form-2

0

0

10

20 30 Time in minutes

40

50

60

Figure-11 In Vitro powder dissolution profile of ceritinib Form-1, 2 and 3. 3.5.2. ICH Stability testing Polymorph stability All the forms of ceritinib were stable at ambient conditions of Hyderabad (35°C and 40% RH) for more than six months. Ceritinib Forms-2 and 3 are subjected to stability testing in accordance with US-FDA stability protocol-Q1A (R2) (Stability stating of new drug substance and products).47 Both Form-2 and Form-3 are subjected to stress [Photo Stress (254 nm/ 365 nm), Accelerated stress(40°C ± 2°C, 75% RH ± 5%RH), Thermal stress (60°C ± 2°c) and compression stress],Conditions, testing intervals and observation for stress study are tabulated in Table-8. The results of stability study indicate that both forms are stable, physically and chemically under above said stress conditions. Hygroscopic study was carried out by exposing the material to a humidity of RH = 90% ± 10% in open conditions at room temperature. Conditions, testing intervals and observation of relative humidity study are tabulated in Table-9.



1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60

Page 26 of 33

From the results, it is evident that none of the phases (Form-2 and 3) show any kind of transformations during the study. Accelerated stability (40°C ± 2°C, 75% RH ± 5%RH), long term stability (25°C ± 2°C, 60% RH ± 5%RH) and low temperature stability (2 to 8°C) in packed condition were conducted for both Form-2 and Form-3. These two forms have shown remarkable stability (Physically and chemically) under the above said conditions for six months (Table-10).

Table-8 Stress study data for ceritinib Form-2 and 3. Form-2 Stress study & storage

Water content

%HPLC

%w/w

purity

Photo Stress

Initial 3rd day

2.59 3.02

99.77 99.41

(254 nm/ 365 nm)

10th day

2.60

98.73

Accelerated Stress (40°C±2°C 75%RH±5RH)

3rd day

3.84

98.35

10th day

3.15

99.78

Thermal Stress (60°C ± 2°c)

3rd day 10th day

3.14 2.21

99.63 98.94

Compressed stress

After 1hr of compression

2.42

99.82

Water content

%HPLC

%w/w

purity

0.62 1.18

99.77

conditions Initial

Testing intervals

PXRD Form-2

Matches with initial Form-2

Form-3 Stress study & storage conditions Initial Photo Stress (254 nm/ 365 nm)

Testing intervals Initial 3rd day

1.02 1.44

98.68 99.60

10th day

0.89

99.66

Thermal Stress (60°C ± 2°c)

3rd day 10th day

0.12 0.79

99.57 98.65

Compressed stress

After 1hr of compression

1.06

99.62


Form-3

99.59

10th day 3rd day

Accelerated Stress (40°C±2°C 75%RH±5RH)

PXRD

Matches with initial Form-3

Page 27 of 33

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60


Table-9 Hygroscopic study for ceritinib Form-2 and 3. Form-2 Open Condition: Exposed to air at Room temperature Water content (%w/w) Solid Phase Stability with PXRD Initial 2.59 Form-2 1st day 4.17 Matches with initial 3rd day 4.07 Matches with initial Open Condition: Exposed to humidity (90%, ±10%) Testing intervals Water content (%w/w) Solid Phase Stability with PXRD Initial 2.59 Form-2 1st day 8.65 Matches with initial 3rd day 7.47 Matches with initial Form-3 Testing intervals

Open Condition: Exposed to air at Room temperature Water content (%w/w) Solid Phase Stability with PXRD Initial 0.62 Form-3 1st day 1.41 Matches with initial 3rd day 1.38 Matches with initial Open Condition: Exposed to humidity (90%, ±10%) Testing intervals Water content (%w/w) Solid Phase Stability with PXRD Initial 0.62 Form-3 1st day 2.45 Matches with initial 3rd day 2.13 Matches with initial Testing intervals



1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60

Table-10: Six-month stability data for ceritinib Form-2 and 3.

Period Initial 1st Month 2nd month 3rd month 6th month Period Initial 1st Month 2nd month 3rd month 6th month Period Initial 15th Day 1st Month 45th day 2nd month 3rd month 6th month

Period Initial 1st Month 2nd month 3rd month 6th month Period Initial 1st Month 2nd month 3rd month 6th month Period Initial 15th Day 1st Month 45th day 2nd month 3rd month 6th month

Form-2 Storage Condition-2 to 8°C (Low temperature stability) %Water content %HPLC purity Solid phase stability by PXRD 1.89 99.84 Form-2 1.96 99.89 1.91 99.79 Matches with initial 1.90 99.78 1.61 99.81 Storage Condition-25°°C ± 2°°C, 60% RH ± 5%RH (Long term stability) %Water content HPLC purity Solid phase stability by PXRD 1.89 99.84 Form-2 1.75 99.88 1.81 99.79 Matches with initial 1.58 99.69 1.09 99.87 Storage Condition-40°°C ± 2°°C, 75% RH ± 5%RH (Accelerated stability) %Water content %HPLC purity Solid phase stability by PXRD 1.89 99.84 Form-2 1.83 99.93 1.78 99.81 1.62 99.83 Matches with initial 1.72 99.79 1.13 99.58 1.09 99.87 Form-3 Storage Condition-2 to 8°C (Low temperature stability) %Water content %HPLC purity Solid phase stability by PXRD 1.27 99.75 Form-3 1.59 99.83 1.55 99.72 Matches with initial 1.18 99.68 1.5 99.88 Storage Condition-25°°C ± 2°°C, 60% RH ± 5%RH (Long term stability) %Water content HPLC purity Solid phase stability by PXRD 1.27 99.75 Form-3 1.79 99.84 1.39 99.75 Matches with initial 1.31 99.68 1.2 99.84 Storage Condition-40°°C ± 2°°C, 75% RH ± 5%RH (Accelerated stability) %Water content %HPLC purity Solid phase stability by PXRD 1.27 99.75 Form-3 1.55 99.89 1.68 99.81 1.71 99.78 Matches with initial 1.56 99.7 1.11 99.49 1.06 99.87


Page 28 of 33

Page 29 of 33

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60


4.0. Conclusions: Two novel forms of ceritinib are identified during the course of polymorph screening. The scientific rationale behind this discovery is clearly nailed down by the integration of experimental and theoretical approach. The anhydrous and hydrate Forms of ceritinib are isolated by following pH modulation crystallization. Both the polymorphs have advantages in terms of stability, solubility and dissolution rate compare to Form-1. The present discovery has demonstrated a new approach of crystallization with scientific rationale. We hope the discovered forms may be useful on the downstream of product development by following appropriate formulation strategies.

Acknowledgements: The authors would like to thank Dr. Reddy’s laboratory for encouraging and providing facility to do fundamental research. We would also like to thank Prof T. N. Guru Row for providing the access to the SXRD data collection.

Supporting information files: The supporting Information is available free of charge on the ACS Publications website. The Crystallographic information files for all crystalline forms can be downloaded from Cambridge structure data base with the CCDC No:1563222, PXRD of Form-1 to 3 are given in Figure S1 to S3; DSC of Form-1 to 3 are given in Figure S4 to S8;TGA of Form-1 to 3 is given in Figure S7 to S9;Moisture content (MC) of Form-1 to 3 are given in Figure S10 to S12;Table-S1: Excess enthalpy calculation for ceritinib with different category of solvents; Table S2: The experimental trails and their results for hydrate screening.



1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60

Page 30 of 33

References: 1. Thayer, A. M. Chem. Eng. News, 2010, 13, 88. 2. Babu, N. J.;Nangia, A. Cryst. Growth Des. 2011, 11, 2662. 3. Berge,S. M.;Bighley, L. D.;Monkhouse, D. C. J. Pharm. Sci., 1977, 66, 1. 4. Serajuddin, A. T. M. Adv. Drug Delivery Rev., 2007, 59, 603. 5. Banerjee, R.; Bhatt, P. M.; Ravindra, N. V.; Desiraju, G. R. Cryst. Growth Des. 2005, 5, 2299. 6.

Thakuria, R.; Nangia, A. CrystEngComm,2011, 13, 1759.

7. Almarsson, Ö.; Zaworotko, M. J. Chem. Commun., 2004, 1889. 8. Cherukuvada, S.; Babu, N. J.; Nangia, A.J. Pharm. Sci., 2011, 100, 3233. 9. Trask, A. V.;Motherwell,W. D. S.;Jones, W.Cryst. Growth Des., 2009, 5, 1013. 10. Thomas, S. P.; Nagarajan, K.;Row, T. N. G.Chem. Commun.,2012, 48, 10559–10561. 11. Swapna, B.; Suresh, K.; Nangia, A. Chem. Commun.,2016, 52, 4037–4040. 12. Joseph, S.; Sathishkumar, R.; Mahapatra, S.; Desiraju, G. R. Acta. Cryst.2011, B67, 525534. 13. Remenar, J. F.; Morissette, S. L.; Peterson, M. L.; Moulton, B.; MacPhee, J. M.;Guzmán, H. R.; Almarsson,Ö. J. Am.Chem. Soc., 2003, 125, 8456. 14. Sanphui, P.; Goud, N. R.; Khandavilli, U. B. R.; Nangia, A.Cryst. Growth Des., 2011, 11, 4135. 15. Banerjee, R.; Bhatt, P. M.; Ravindra, N. V.; Desiraju, G. R. Cryst. Growth Des., 2005, 5, 2299. 16. Good,D. J.; Rodríguez-Hornedo, N. Cryst. Growth Des., 2009, 9, 2252. 17. Chennuru, R.; Muthudoss, P.; Voguri, R. S.; Ramakrishnan, S.; Peddy V.;Babu, R. R. C.;Mahapatra, S. Cryst. Growth Des., 2016, 17(2), 612-628. 18. Long, S. H.; Zhou, P. P.; Theiss, K. L.; Siegler, M. A.; L.Li, T.CrystEngComm, 2015, 17, 5195–5205. 19. Chennuru, R.; Muthudoss, P.; Ramakrishnan , S.; Mohammad, A. B.; Babu, R. R. C.; Mahapatra , S.; Nayak, S. K. Journal of Molecular Structure, 2016, 86, 1120. 20. Kulkarni, C.; Kelly, A.; Kendrick, J.; Gough, T.; Paradkar, A. Cryst. Growth Des., 2013, 13, 5157–5161.


Page 31 of 33

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60


21. Mahapatra, S.; Nayak, K. S.; Prathapa, S. J.; Guru Row, T. N. Crystal Growth and Design, 2008, 8(4), 1223. 22. Trask, A. V.; Shan, N.; Motherwell, W. D.; Jones, W.; Feng, S.; Tan, R. B.; Carpenter, K. J. Chem. Commun.,2005, 880–882. 23. Maher, A.; Croker, D. M.; Seaton, C. C.; Rasmuson, Å. C.; Hodnett, B. K. Cryst. Growth Des., 2014, 14, 3967–3974. 24. Girard, J.; Fromm, K. CrystEngComm, 2012, 14,6487–6491. 25. Mahapatra, S.; Azim.Y.; Desiraju, G. R. journal of molecular structure. 2010, 976(13),200. 26. Eckert, F.; Klamt, A. AIChE J.2002, 48 (2), 369–385. 27. Eckert, F.; Klamt, A. KG, Leverkusen, Ger.2006. 28. Klamt, A. Wiley Interdiscip. Rev. Comput. Mol. Sci.2011, 1 (5), 699–709. 29. Klamt, A. COSMO-RS: From Quantum Chemistry to Fluid PhaseThermodynamics and Drug Design; Elsevier, 2005. 30. Campeta, A. M.; Chekal, B. P.; Abramov, Y. A.; Meenan, P. A.; Henson, M. J.; Shi, B.; Singer, R. A.; Horspool, K. R. J. Pharm. Sci.2010, 99 (9), 3874–3886. 31. Newman, A. Org. Process Res. Dev.2013, 17 (3), 457–471. 32. Cui, Y.; Yao, E. J. Pharm. Sci. 2007, 97, 2730. 33. Sistla, A.; Wu, Y.; Khamphavong, P.; Liu, J. Pharm. Dev. Technol.2011, 16, 102. 34. Ten Brink, T.; Exner, T. E. J. Comput. Aided. Mol. Des.2010, 24 (11), 935–942. 35. Etter, M. C. Acc. Chem. Res. 1990, 23, 120–126. 36. Boultif, A.; Louër, D. J. Appl. Crystallogr.1991, 24 (6), 987–993. 37. Neumann, M. A. J. Appl. Crystallogr.2003, 36 (2), 356–365. 38. Boultif, A.; Louër, D. J. Appl. Crystallogr.1991, 24 (6), 987–993. 39. Pawley, G. S. J. Appl. Cryst. 1981, 14, 357-361. 40. Stahly, G. P. Cryst. Growth Des.2007,7, 1007. 41. Vippagunta, S. R.; Brittain, H. G.; Grant, D. J. W. Adv. Drug Deliv. Rev.2001, 48 (1), 3– 26. 42. Ford, J. L.; Timmins, P. Pharmaceutical thermal analysis: techniques and applications; Halsted Press, 1989. 43. Grzesiak, A. L.; Lang, M.; Kim, K.; Matzger, A. J. J. Pharm. Sci. 2003, 92, 2260.



1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60

44. Stuart, B. Infrared spectroscopy; Wiley Online Library, 2005. 45. Smith, E.; Dent, G. Modern Raman spectroscopy: a practical approach; John Wiley & Sons, 2013. 46. Bugay, D. E.; Brittain, H. G.; Cogdill, R. P.; Drennen, J. K.; Brittain, H. Taylor & Francis, New York 2006. 47. US-FDA stability protocol-Q1A (R2) -Industry, G. for. Ich 2003, No. November, 1–22.


Page 32 of 33

Page 33 of 33

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60


TABLE OF CONTENTS USE ONLY

In-Situ Metastable Form: A Route For The Generation Of Hydrate And Anhydrous Form Of Ceritinib Ramanaiah Chennuru#$, Ravi Teja Koya#, Pavan Kommavarapu#, Saladi Venkata Narasayya#,Prakash Muthudoss#, R. Ravi Chandra Babu$* and Sudarshan Mahapatra#*

Synopsis: Two novel forms of ceritinib are identified during the course of polymorph screening. The scientific rationale behind this discovery is clearly nailed down by the integration of experimental and theoretical approach. The anhydrous and hydrate Forms of ceritinib are isolated by following pH modulation crystallization. Both the polymorphs have advantages in terms of stability, solubility and dissolution rate compare to Form-1. The present discovery has demonstrated a new approach of crystallization with scientific rationale.


In Situ Metastable Form: A Route for the Generation of Hydrate and

Recommend Documents