Cambrex Profarmaco Milano S.r.l.

Cambrex Profarmaco Milano S.r.l.

EMTRICITABINE
ACTIVE SUBSTANCE MASTER FILE

Ed. 01.00/2012-ASMF
NEW SUBMISSION OCTOBER 2012
EMTRICITABINE - DOSSIER Ed. 01.00/2012-ASMF INDEX (CTD - FORMAT)

MODULE 1 - ADMINISTRATIVE INFORMATION AND PRESCRIBING INFORMATION 1.4.1 Annex I Information about the expert quality Environmental risk assessment
MODULE 2 - COMMON TECHNICAL DOCUMENT SUMMARIES 2.3 2.3 Quality overall summary (AP) Quality overall summary (RP)*
MODULE 3 - QUALITY 3.2.S SECTION A 3.2.S.1 3.2.S.1.1 3.2.S.1.2 DRUG SUBSTANCE GENERAL INFORMATION Nomenclature Structure Structural formula Molecular formula Molecular weight 3.2.S.1.3 General properties A 1/3 A-2 A-2 A-2 A-2 A-2 A-3
SECTION B1
3.2.S.2
MANUFACTURE 3.2.S.2.1 Manufacturer 3.2.S.2.2 Description of manufacturing process and process controls 3.2.S.2.2.1 Scheme of the synthesis 3.2.S.2.2.2 Scheme of the manufacturing process* 3.2.S.2.2.3 Brief description of the synthesis* 3.2.S.2.2.4 Flow-sheet* 3.2.S.2.2.5 Detailed description of the manufacturing process* 3.2.S.2.2.6 Reprocessing* 3.2.S.2.2.7 In-process controls*
B1 1/30 B1 - 2 B1 - 3 B1 - 3 B1 - 4 B1 5 B1 6 B1 - 8 B1 - 20 B1 - 21 B2 1/17 B2 - 2 B2 - 3 B2 - 3 B2 - 5 B2 - 16 B2 - 17
SECTION B2
3.2.S.2.3 Control of materials* 3.2.S.2.3.1 Raw materials list* 3.2.S.2.3.2 Raw materials sampling procedure* 3.2.S.2.3.3 Raw materials specifications and test methods* 3.2.S.2.4 Control of critical steps and intermediates* 3.2.S.2.5 Process validation* 3.2.S.2.6 Manufacturing process development*
Emtricitabine, 10/2012 Ed 01.00/2012-ASMF
Index - 1/3
SECTION C
3.2.S.3
CHARACTERIZATION 3.2.S.3.1 Elucidation of structure and other characteristics 3.2.S.3.1.1 Spectral analysis - NMR Spectrum - IR Spectrum - Mass Spectrum - Elemental analysis 3.2.S.3.1.2 Potential isomerism 3.2.S.3.1.3 Polymorphism 3.2.S.3.2 Impurities 3.2.S.3.2.1 Related substances and degradation products 3.2.S.3.2.2 Description of impurities 3.2.S.3.2.3 Characterization of impurities
C 1/73 C-2 C-2 C-2 C - 12 C - 16 C - 25 C - 28 C - 30 C - 43 C - 43 C - 52 C - 55
SECTION D
3.2.S.4
CONTROL OF DRUG SUBSTANCE 3.2.S.4.1 3.2.S.4.2 3.2.S.4.3 3.2.S.4.4 3.2.S.4.5 Specification Analytical procedures Validation of analytical procedures Batch analysis Justification of specification
D 1/7 D-2 D-2 D-2 D-3 D-7
SECTION E
3.2.S.5
REFERENCE STANDARD
E 1/3
SECTION F
3.2.S.6
CONTAINER CLOSURE SYSTEM 3.2.S.6.1 Specifications 3.2.S.6.2 Labeling
F 1/45 F-3 F - 45
SECTION G
3.2.S.7
STABILITY 3.2.S.7.1 Stability summary and conclusions 3.2.S.7.1.1 General stability protocol 3.2.S.7.1.2 Summary of long term stability studies 3.2.S.7.1.3 Summary of accelerated stability studies 3.2.S.7.1.4 Summary of stress stability studies 3.2.S.7.2 Post approval stability protocol and stability commitment 3.2.S.7.3 Stability data 3.2.S.7.3.1 Stability specifications 3.2.S.7.3.2 Long term stability studies 3.2.S.7.3.3 Accelerated stability studies 3.2.S.7.3.4 Stress stability studies
G 1/115 G-2 G-2 G-4 G-4 G-4 G-5 G-6 G-6 G-7 G-7 G-9
Index - 2/3
ATTACHMENTS Attachment 1 Section D Attachment 2 Section D Attachment 3 Section B2 Attachment 4 Section G Attachment 5 Section D Attachment 6 Section D Attachment 7 Section D Attachment 8 Section D Attachment 9 Section B2 *) Restricted part Specifications Analytical procedures Raw material specifications* Stability specifications and Stability data Copper determination by atomic absorption - validation Validation report for assay by HPLC Enantiomeric purity and chiral identity by hplc-Validation Related substances by HPLC Validation Starting material qualification*
Index - 3/3
Module 1 - Annex I ENVIRONMENTAL RISK ASSESSMENT
Annex I - Environmental risk assessment - 1/3
ENVIRONMENTAL RISK ASSESSMENT
Name of the applicant: CAMBREX PROFARMACO MILANO S.r.l. Address: Registered Office and Manufacturing Facility Cambrex Profarmaco Milano S.r.l. Via Curiel, 34 20067 Paullo (Milano) Italy
Identification of chemical substance EMTRICITABINE

O O HO S N F N NH2
Molecular weight: 247.3 Molecular formula: C8H10FN3O3S Description Cambrex Profarmaco Milano S.r.l. is seeking approval for a Dossier to manufacture EMTRICITABINE at its facility located in Paullo, a small town in the hinterland of Milan, Italy. EMTRICITABINE is intended for use in human health and will be distributed as a powder in bulk. During the manufacturing process we will generate three different types of wastes: low contaminated water, highly contaminated liquids and gaseous emissions. All these wastes will be treated on site: - the wastewater in our ecological department using a typical chemical, physical and biological process, from which we generate clean water and a sludge classified as non hazardous solid waste that is disposed off-site by authorised companies. - the highly contaminated liquids will be burned in our state of the art incinerator. - all gaseous emissions will be washed through a scrubber system and sent to the incinerator as combustion air. Our production site is subject to Italian safety, health and environmental laws and regulations enforced by different state and local authorities:
D. Lgs. 81 of 9/4/2008 D.L. 334 of 17/8/1999 A.I.A./IPPC of 7/9/2007 D.P.R. 203 of 24/5/1988 D.L. 152 of 11/5/1999 D.L. 22 of 5/2/1997 94/67CE of 16/12/1994
(health and safety) (risk activities) (environmental integrated authorization of the site) (air) (water) (wastes) (incineration of hazardous wastes)
Cambrex Profarmaco Milano is in compliance with all environmental laws and believes that the approval of this Dossier will have not effect on the current air emission permits or wastewater treatment parameters. Cambrex Profarmaco Milano is in compliance with all occupational, safety and health laws as well, and believes that the approval of this Dossier will have no significant effects on the current situation. The production of EMTRICITABINE has been assessed from a safety point of view.
Fate of emitted substances in the environment The liquid wastes and gaseous emissions are burned in our incinerator in compliance with the Italian law; the composition of the emitted air is monitored regularly by an external lab and the local authority, and is constantly within the enforced limits. The non hazardous solid waste generated by the wastewater treatment plant is disposed off-site by authorised companies.
Environmental effects of released substances We don't see any significant effects on the environmental quality related to the production of EMTRICITABINE
Mitigation measures Cambrex Profarmaco Milano is well equipped to prevent safety and ecological effects on the environment. The all facility has been designed and built according to the Italian laws and to the good engineering practices. In our organisation we have dedicated staff working full time on safety and environmental issues. Cambrex Profarmaco Milano follows the Responsible Care principles. We think the production of EMTRICITABINE will not require the implementation of additional mitigation measures beyond the ones we have already in place.
MODULE 1 ADMINISTRATIVE INFORMATION AND PRESCRIBING INFORMATION
1.4.1 - INFORMATION ABOUT THE EXPERT QUALITY ANNEX I - ENVIRONMENTAL RISK ASSESSMENT
Module 1 - Information about the expert - 1/3
1.4.1 INFORMATION ABOUT THE EXPERT - QUALITY
Name of the expert: Address:
Dr. Fulvio Leone Piselli Via Sangro, 11 Milan ITALY
EMTRICITABINE is not described in any pharmacopeias.
All analytical specifications and test methods are sufficient to control the purity of the drug substance. The specifications for the starting material used in the synthesis route are proper for the control of the quality. Data from the analysis of all scale production batches indicate the ability to manufacture the drug substance complying with the specifications. Stability studies on the product manufactured by Cambrex Profarmaco Milano are on going. I consider the quality, the specifications and the analytical procedures employed completely satisfactory.
Curriculum Vitae
Dott. FULVIO LEONE PISELLI Born Address Employed Milan, November 9 1933 Via Sangro 11 , Milan ITALY Profarmaco from January 1958 to December 1999 Currently as Technical Advisor
1957 Industrial Chemistry Degree Universit degli Studi di Milano 1958 Employed as Responsible of Laboratories: - Research and Development - Quality Control 1969 Manager of Research and Development and Quality Control 1983 Manager of Research & Development , Quality Control, Quality Assurance, Regulatory Affairs, Public Relations. 1991 Profarmaco Technical Director Manager of Health, Safety, Environment 1994 Technical Director Responsible of Service Prevention and Protection 2000 Retirement Technical Advisor
MODULE 2 COMMON TECHNICAL DOCUMENT SUMMARIES
2.3 - QUALITY OVERALL SUMMARY
APPLICANT PART
APPLICANT PART
Module 2 QOS - 1/12
2.3 QUALITY OVERALL SUMMARY APPLICANT PART

2.3.S DRUG SUBSTANCE 2.3.S.1 GENERAL INFORMATION
EMTRICITABINE
O O HO S N F N NH2
Molecular formula: C8H10FN3O3S Molecular weight:

Chemical name:
247.3
(2R,5S)-4-Amino-5-fluoro-1-(2-hydroxymethyl-[1,3]oxathiolan-5-yl)-1H-pyrimidin-2-one (IUPAC) (2R-cis)-4-Amino-5-fluoro-1-[2-(Hydroxymethyl)-1,3-oxathiolan-5-yl]-2(1H)-pyrimidinone (USP) 5-fluoro-1-[(2R,5S)-2-(Hydroxymethyl)-1,3-oxathiolan-5-yl]cytosine (USP)
C.A.S Registry N Cambrex Profarmaco Milano code
143491-57-0 194
APPLICANT PART
Module 2 QOS - 2/12
General properties
Physical Appearance Emtricitabine is a white or almost white crystalline powder.
Solubility Soluble in Water and Methanol Soluble in wet Acetone (KF 5%) 2% at room temperature Sparingly soluble in Chloroform, Methylene chloride and Acetone.
Melting range Emtricitabine melts at about 136-140C (Merck index). Optical rotation []D25 = - 133.60 (c = 0.23 in Methanol) (Merck index)
APPLICANT PART
Module 2 QOS - 3/12
2.3.S.2 MANUFACTURE
EMTRICITABINE is supplied by:
Cambrex Profarmaco Milano S.r.l Via Curiel, 34 20067 Paullo (Milan Italy)
Starting materials The starting materials for the synthesis of Emtricitabine are: FLUOCY (5-Fluorocytosine) HYDOXA
Both starting materials are outsourced and the analytical specifications included in Attachment 3. The qualification documentation for the starting materials is reported in Attachment 9. Final purification solvent: Isopropyl alcohol
APPLICANT PART
Module 2 QOS - 4/12
Scheme of the synthesis of EMTRICITABINE (paragraph 3.2.S.2.2.1)
O O S
HYDOXA
O OH
O HN
NH2
F
FLUOCY
SOCl2
Hexamethyldisilazan
O O O S
Chloro solution
N N
H N Si F
O Cl
Si
Silylcytosine solution
Diisopropylethylamine
O O HO S N
NH2
1) NaBH4 2) Maleic Acid
O O S O
O N
NH2
F
(Maleate salt 1:1) FTC maleate
FTC Menthylester
Triethylamine Isopropanol
O O HO S N
NH2
XX Wet isopropanol CRYSTALLIZED EMTRICITABINE (FTC BASE)
EMTRICITABINE (FTC BASE)
APPLICANT PART
Module 2 QOS - 5/12
2.3.S.3 CHARACTERIZATION
Evidence of chemical structure (paragraph 3.2.S.3.1.1) The elucidation of the chemical structure of Emtricitabine is based on 1H-NMR, 13C-NMR, MS and IR spectra, and on Elemental Analysis data. Potential isomerism (paragraph 3.2.S.3.1.2) The molecule has two stereocenters indicated in the picture below. The isomer of interest is the cis2R,5S. The specifications limits for all the isomers (epimers and enantiomer) are reported in the analytical specifications conform to US pharmacopoeia (pending draft) . The limits for all the isomers by HPLC are : NMT 0.3% Emtricitabine Enantiomer NMT 0.2% Emtricitabine 1st 5-EPIMER + Emtricitabine 2nd 5-EPIMER
Polymorphism (paragraph 3.2.S.3.1.3) The study has demonstrated that Cambrex Profarmaco Milano current production of Emtricitabine is consistent from the point of view of the polymorphic form: the IR spectra, the DSC thermograms and the X-ray diffraction patterns are perfectly superimposable. See paragraph 3.2.S.3.1.3 for the detailed data.
APPLICANT PART
Module 2 QOS - 6/12
Specified identified impurities of EMTRICITABINE (paragraph 3.2.S.3.2) The specified impurities for Emtricitabine are the following: 1) MALEIC ACID 2) 4-Amino-5-fluoropyrimidin-2-(1H)-one (FLUOROCYTOSINE) 3) cis-5-[4-amino-5-fluoro-2-oxopyrimidin-1(2H)-yl]-1,3-oxathiolane-2-carboxylic acid (EMTRICITABINE ACID) 4) 5-Fluoro-1-[(2R,3RS,5S)-2-(hydroxymethyl)-3-oxo-1,3-oxathiolan-5-yl]cytosine (EMTRICITABINE SULFOXADES) 5) (2)(-)-1-[(2R,5S)-2-(Hydroxymethyl)-1,3- oxathiolan-5-yl]cytosine (LAMIVUDINE) 6) 5-Fluoro-1-[(2R,5S)-2-(hydroxymethyl)-1,3-oxathiolan-5-yl]uracil (FLUOROURACIL ANALOG) 7) L-Menthyl-5-(2R,5S)-[4-amino-5-fluoro-2-oxopyrimidin-1(2H)-yl]-1,3-oxathiolane2-carboxylate (MENTHYL ESTER)
APPLICANT PART
Module 2 QOS - 7/12
2.3.S.4 CONTROL OF THE DRUG SUBSTANCE
Specification for EMTRICITABINE (paragraph 3.2.S.4.1):
APPLICANT PART
Module 2 QOS - 8/12
Justification of specifications (paragraph 3.2.S.4.5) The Active Ingredient Emtricitabine is not described in any Pharmacopoeia. All the tests included in the Specification for Final Product have been developed internally by Cambrex Profarmaco.
Batch analysis (paragraph 3.2.S.4.4) The certificates of analysis of three news standard production batches are reported in Paragraph 3.2.S.4.4. The results enclosed show that EMTRICITABINE complies with the specifications for the final product.
APPLICANT PART
Module 2 QOS - 9/12
2.3.S.5 REFERENCE STANDARD
EMTRICITABINE Working Standard: lot N 140512 Emtricitabine WS has been prepared by Cambrex Profarmaco R& D laboratory. It is a standard manufacturing batch produced according to the manufacturing process described in section 3.2.S.2.2 Manufacture and is maintained at room temperature. Emtricitabine working standard was not additionally re-crystallized. The characterisation of the working standard has been presented in paragraph 3.2.S.3.1.1 Spectral analysis. The characterisation of the working standard has been presented in paragraph 3.2.S.3.1.1 Spectral analysis. The certificate of analysis is included is reported in Paragraph 3.2.S.5.
2.3.S.6 CONTAINER CLOSURE SYSTEM
The commercial packaging system for EMTRICITABINE, composed by plastic drums, closure systems, polyethylene bags, is fully-described in this section. The specifications of the suppliers, reporting the general characteristics of drums and polyethylene bags and certifying their food-grade, are enclosed. The specifications of the commercial packaging system are enclosed in Paragraph 3.2.S.6.1.
APPLICANT PART
Module 2 QOS - 10/12
2.3.S.7 STABILITY
General stability protocol: For the general protocol for stability studies see paragraph 3.2.S.7.1.1 For the post-approval stability protocol see paragraph 3.2.S.7.2. Stability specifications (paragraph 3.2.S.7.3.1):
These specifications are used to test the product during the Accelerated and the Long term stability studies. The tests methods refer to the release specifications for the active ingredient.
APPLICANT PART
Long term stability studies (paragraph 3.2.S.7.3.2)

Long term stability studies of EMTRICITABINE are on-going on the following batches:
Batches 010102 010304 010506 Batch size (Kg) 25 Kg 32 Kg 34 Kg Date of manufacture 12/2012 12/2012 12/2012
These lots have been stored under the following conditions:

LONG TERM STABILITY (LTS)
Temperature Relative humidity 30C 2C 75% 5%
Accelerated stability studies (paragraph 3.2.S.7.3.3)

Accelerated stability studies of EMTRICITABINE are on-going on the following batches:

ACCELERATED STABILITY (AS)
APPLICANT PART
MODULE 3 - QUALITY
3.2.S - DRUG SUBSTANCE
Module 3
3.2.S.1 GENERAL INFORMATION
Section A
3.2.S.1
GENERAL INFORMATION 3.2.S.1.1 3.2.S.1.2 Nomenclature Structure 3.2.S.1.3 Structural formula Molecular formula Molecular weight
General properties
General information A - 1/3
3.2.S.1.1 - NOMENCLATURE
Chemical name (2R,5S)-4-Amino-5-fluoro-1-(2-hydroxymethyl-[1,3]oxathiolan-5-yl)-1H-pyrimidin-2-one (IUPAC) (2R-cis)-4-Amino-5-fluoro-1-[2-(Hydroxymethyl)-1,3-oxathiolan-5-yl]-2(1H)-pyrimidinone (USP) 5-fluoro-1-[(2R,5S)-2-(Hydroxymethyl)-1,3-oxathiolan-5-yl]cytosine (USP)
C.A.S Registry N Cambrex Profarmaco Milano code
143491-57-0 194
3.2.S.1.2 - STRUCTURE
Structural formula
O O HO S N F N NH2
Molecular formula: C8H10FN3O3S Molecular weight: 247.3
Therapeutic category: Antiviral
3.2.S.1.3 - GENERAL PROPERTIES
Physical Appearance Emtricitabine is a white or almost white crystalline powder.
Solubility Soluble in Water and Methanol Soluble in wet Acetone (KF 5%) 2% at room temperature Sparingly soluble in Chloroform, Methylene chloride and Acetone.
Melting range Emtricitabine melts at about 136-140C (Merck index). Optical rotation []D25 = - 133.60 (c = 0.23 in Methanol) (Merck index)
3.2.S.2 - MANUFACTURE
Section B1
3.2.S.2 MANUFACTURE 3.2.S.2.1 3.2.S.2.2 Manufacturer Description of manufacturing process and process controls 3.2.S.2.2.1 Scheme of the synthesis 3.2.S.2.2.2 Scheme of the manufacturing process * 3.2.S.2.2.3 Brief description of the synthesis* 3.2.S.2.2.4 Flow-sheet* 3.2.S.2.2.5 Detailed description of the manufacturing process * 3.2.S.2.2.6 Reprocessing* 3.2.S.2.2.7 In-process controls*
Section B2
3.2.S.2.3
Control of materials* 3.2.S.2.3.1 Raw materials list* 3.2.S.2.3.2 Raw materials sampling procedure* 3.2.S.2.3.3 Raw materials specifications and test methods*
3.2.S.2.4 3.2.S.2.5 3.2.S.2.6
Control of critical steps and intermediates* Process validation* Manufacturing process development*
Attachments list of Section B2 Attachment 3 Attachment 9 Raw materials specifications* Starting material qualification*
* Restricted part
Manufacture B1 - 1/30
SECTION B1
3.2.S.2.1 - MANUFACTURER
ADMINISTRATIVE ADDRESS Cambrex Profarmaco Milano S.r.l. Via Accademia 29 20131 Milano (Italy) Telephone: +39 02 345988.1 Fax: +39 02 33105730 / +39 33105606 REGISTERED OFFICE AND MANUFACTURING FACILITY Cambrex Profarmaco Milano S.r.l. Via Curiel, 34 20067 Paullo, Milano Italy Telephone : +39 02 9062601 Fax : +39 02 90630995 RESPONSIBLE OFFICIAL Dr. Paolo Russolo President Via Curiel, 34 20067 Paullo, Milano Italy US AGENT Mr. Hal Lipton GYMA LABORATORIES of AMERICA INC. 135 Cantiague Rock Road Westbury, N.Y. 11590 IN-HOUSE STABILITY TESTING LABORATORY Cambrex Profarmaco Milano S.r.l Via Curiel, 34 20067 Paullo (Milano) Italy ALTERNATIVE STABILITY TESTING LABORATORY (*) PRC (Pharma Research Centre) TICINUM LAB Via Bovio, 6 28100 NOVARA, Italy
(*) The description of the external stability testing laboratory is included in Attachment 5.
3.2.S.2.2 - DESCRIPTION OF MANUFACTURING PROCESS AND PROCESS CONTROLS

3.2.S.2.2.1 Scheme of the synthesis EMTRICITABINE
O O S
HYDOXA
O OH
O HN
NH2
F
FLUOCY
SOCl2
Hexamethyldisilazan
O O O S
Chloro solution
N N
H N Si F
O Cl
Si
Silylcytosine solution
Diisopropylethylamine
O O HO S N
NH2
1) NaBH4 2) Maleic Acid
O O S O
O N
NH2
F
(Maleate salt 1:1) FTC maleate
FTC Menthylester
Triethylamine Isopropanol
O O HO S N
NH2
XX Wet isopropanol CRYSTALLIZED EMTRICITABINE (FTC BASE)
EMTRICITABINE (FTC BASE)
3.2.S.3 - CHARACTERIZATION
Section C
3.2.S.3
CHARACTERIZATION 3.2.S.3.1 Elucidation of structure and other characteristics 3.2.S.3.1.1 Spectral analysis - NMR Spectrum - IR Spectrum - Mass Spectrum - Elemental analysis 3.2.S.3.1.2 Potential isomerism 3.2.S.3.1.3 Polymorphism 3.2.S.3.2 Impurities 3.2.S.3.2.1 Related substances and degradation products 3.2.S.3.2.2 Description of impurities 3.2.S.3.2.3 Characterization of impurities
Characterization
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3.2.S.3.1 - ELUCIDATION OF STRUCTURE AND OTHER CHARACTERISTICS

3.2.S.3.1.1 Spectral analysis The elucidation of the chemical structure of Emtricitabine is based on 1H-NMR, 13C-NMR, MS and IR spectra and on Elemental Analysis data. NMR SPECTRA Nuclear Magnetic Resonance spectra 1H and 13C of Emtricitabine WS .
Characterization
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Characterization
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Characterization
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Characterization
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Characterization
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Characterization
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Characterization
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Characterization
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Characterization
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Characterization
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IR SPECTRUM The FT-IR spectrum of Emtricitabine WS was recorded on a JASCO FT-IR 420 Spectrometer.
Characterization
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Characterization
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Characterization
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Characterization
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MS SPECTRUM The MS analysis of Emtricitabine WS was performed using an Agilent 6310. The fragmentation peaks conform to the proposed structure.
Characterization
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Characterization
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Characterization
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Characterization
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Characterization
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Characterization
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Characterization
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Characterization
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Characterization
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ELEMENTAL ANALYSIS The results of the elemental analysis of Emtricitabine WS agree with the proposed structure, as reported in the following table.
Characterization
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Characterization
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Characterization
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3.2.S.3.1.2 Potential Isomerism
The molecule has two stereocenters indicated in the picture below. The isomer of interest is the cis2R,5S. The specifications limits for all the isomers (epimers and enantiomer) are reported in the analytical specifications conform to US pharmacopoeia (pending draft) . The limits for all the isomers by HPLC are : NMT 0.3% Emtricitabine Enantiomer NMT 0.2% Emtricitabine 1st 5-EPIMER + Emtricitabine 2nd 5-EPIMER
Characterization
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3.2.S.3.1.3 Polymorphism
Characterization
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Characterization
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Characterization
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Characterization
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Characterization
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Characterization
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Characterization
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Characterization
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Characterization
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Characterization
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Characterization
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Characterization
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Characterization
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Characterization
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Characterization
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Characterization
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Characterization
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Characterization
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Characterization
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Characterization
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Characterization
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3.2.S.3.2 - IMPURITIES
3.2.S.3.2.1 Related substances and degradation products
The specified impurities for Emtricitabine are the following: 1) MALEIC ACID 2) 4-Amino-5-fluoropyrimidin-2-(1H)-one (FLUOROCYTOSINE) 3) cis-5-[4-amino-5-fluoro-2-oxopyrimidin-1(2H)-yl]-1,3-oxathiolane-2-carboxylic acid (EMTRICITABINE ACID) 4) 5-Fluoro-1-[(2R,3RS,5S)-2-(hydroxymethyl)-3-oxo-1,3-oxathiolan-5-yl]cytosine (EMTRICITABINE SULFOXADES) 5) (2)(-)-1-[(2R,5S)-2-(Hydroxymethyl)-1,3- oxathiolan-5-yl]cytosine (LAMIVUDINE) 6) 5-Fluoro-1-[(2R,5S)-2-(hydroxymethyl)-1,3-oxathiolan-5-yl]uracil (FLUOROURACIL ANALOG) 7) L-Menthyl-5-(2R,5S)-[4-amino-5-fluoro-2-oxopyrimidin-1(2H)-yl]-1,3-oxathiolane2-carboxylate (MENTHYL ESTER)
The standard HPLC purity profile for Emtricitabine manufactured by Cambrex Profarmaco Milano is the following:
Characterization
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Residual solvents The solvents used during the synthesis of Emtricitabine are:
Isopropyl alcohol (Solvent of purification) Acetone Acetonitrile Methylene chloride Dimethyldormammide Methylisobutylketone Toluene
Only Isopropyl alcohol included in the release certificate of analysis.
Characterization
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Metal catalysts In the process are not used metal catalysts.
Characterization
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3.2.S.3.2.2 - Description of impurities
1) cis-butenedioic acid MALEIC ACID
This substance [as Raw material] is utilized in the process to isolate crude FTC as the Maleate salt during the synthesis of Emtricitabine. Maleic acid is removed as triethylamonium salt by treatment with triethylamine.
2) 4-Amino-5-fluoropyrimidin-2(1H)-one FLUOROCYTOSINE
N HN NH2 F
This impurity is the starting material of the manufacturing process. It was never observed during the stress stability studies (see paragraph 3.2.S.7.3.4) and therefore it is not considered a potential degradation impurity.
3) cis-5-[4-amino-5-fluoro-2-oxopyrimidin -1(2H)-yl]-1,3-oxathiolane-2-carboxylic acid EMTRICITABINE ACID

O HO O O S N F N NH2
This impurity could originate through hydrolysis of FTC Menthylester in the synthesis of Emtricitabine due to the moderate alkaline reaction conditions. It is generally not observed, probably because the reaction mixture is buffered to pH < 9.5.
Characterization
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4) 5-Fluoro-1-[(2R,3RS,5S)-2-(hydroxymethyl)-3-oxo-1,3-oxathiolan-5-yl] cytosine EMTRICITABINE SULFOXIDES

O HO O S O
+
N N
NH2 F
This is a potential degradation impurity that is observed in oxidative stress and, to a lesser extent, in acidic and alkaline stress.
5) (2)(-)-1-[(2R,5S)-2-(Hydroxymethyl)-1,3-oxathiolan-5-yl]cytosine LAMIVUDINE
O HO O S N N NH2
This impurity can originate through replacement of a fluorine atom by an hydride ion in the synthesis of Emtricitabine (reduction of FTC-Menthylester by sodium borohydride).
Characterization
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6) 5-Fluoro-1-[(2R,5S)-2-(hydroxymethyl)-1,3-oxathiolan-5-yl]uracil FLUOROURACIL ANALOG
O HO O S N
H N
O F
This is a potential degradation impurity observed in acidic and alkaline stress.
7) L-Menthyl 5-(2R,5S)-[4-amino-5-fluoro-2-oxopyrimidin-1(2H)-yl]-1,3-oxathiolane-2carboxylate MENTHYL ESTER
O O O O S N
NH2 F
This substance is an intermediate of the process, isolated in the synthesis of Emtricitabine. Its carry over appeared to be very low.
Characterization
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3.2.S.3.2.3 - Characterization of impurities
The impurities Maleic acid, Fluorocytosine, Lamivudine, are purchased and the relevant certificates of analysis are included in the following pages. The impurities Emtricitabine Acid and Menthyl Ester have been characterized as follows : Emtricitabine Sulfoxides and Fluorouracil Analog standards are not available from the market nor from the Pharmacopoeia. According to USP method, these impurities are identified by retention time.
Characterization
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Emtricitabine Acid
O HO O O S N
NH2 F
The elucidation of the chemical structure of the impurity Emtricitabine acid is based on 1H-NMR.
Characterization
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Characterization
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Characterization
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Characterization
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L-Menthyl 5-(2R,5S)-[4-amino-5-fluoro-2-oxopyrimidin-1(2H)-yl]-1,3-oxathiolane2-carboxylate ( MENTHYL ESTER)
O O O O S N
NH2 F
The elucidation of the chemical structure of the impurity Menthyl ester is based on 1H-NMR,
Characterization
C - 61/73
Characterization
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Characterization
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Characterization
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Characterization
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cis-butenedioic acid MALEIC ACID
Characterization
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Characterization
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Characterization
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Characterization
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4-Amino-5-fluoropyrimidin-2-(1H)-one
(FLUOROCYTOSINE)
Characterization
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Characterization
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(2)(-)-1-[(2R,5S)-2-(Hydroxymethyl)-1,3-oxathiolan-5-yl]cytosine LAMIVUDINE
O HO O S N
NH2
Characterization
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Characterization
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3.2.S.4 - CONTROL OF DRUG SUBSTANCE
Section D
3.2.S.4
CONTROL OF DRUG SUBSTANCE 3.2.S.4.1 3.2.S.4.2 3.2.S.4.3 3.2.S.4.4 3.2.S.4.5 Specification Analytical procedures Validation of analytical procedures Batch analysis Justification of specification
Attachments list of Section D Attachment 1 Attachment 2 Attachment 6 Attachment 7 Attachment 8 Specifications Analytical procedures Assay by HPLC - validation Enantiomeric purity and chiral identity by hplc Related substances by HPLC Validation
Control of drug substance
D - 1/7
3.2.S.4.1 - SPECIFICATIONS
The specifications for the analysis of Emtricitabine are reported in Attachment 1.
3.2.S.4.2 - ANALYTICAL PROCEDURES

The Analytical procedures for Emtricitabine are reported in Attachment 2.
3.2.S.4.3 - VALIDATION OF ANALYTICAL PROCEDURES

The HPLC Assay method has been developed in-house and validated. The HPLC Related Substances method has been developed in-house and validated.
All these methods are routinely used on all batches.
The validation data are reported in the following attachments:

Validation report for assay by HPLC Enantiomeric purity and chiral identity by hplc Validation Related substances by HPLC Validation Attachment 6 Attachment 7 Attachment 8
D - 2/7
3.2.S.4.4 - BATCH ANALYSIS

The standard batch size of the final API is about 35 Kg. We attach the analysis of the three validation batches representative of the standard production of Emtricitabine. EMTRICITABINE
Lot number Manufacturing date Batch size 010102 12/2012 25 Kg 010304 12/2012 32 Kg 010506 12/2012 34 Kg
D - 3/7
D - 4/7
D - 5/7
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3.2.S.4.5 - JUSTIFICATION OF SPECIFICATION

The Active Ingredient Emtricitabine is not described in any Pharmacopoeia. All the tests included in the Specification for Final Product have been developed internally by Cambrex Profarmaco.
D - 7/7
3.2.S.5 - REFERENCE STANDARD
Section E
Reference standard
E - 1/3
3.2.S.5 - REFERENCE STANDARD
EMTRICITABINE Working Standard: lot N 140512 Emtricitabine WS has been prepared by Cambrex Profarmaco R& D laboratory. It is a standard manufacturing batch produced according to the manufacturing process described in section 3.2.S.2.2 Manufacture and is maintained at room temperature. Emtricitabine working standard was not additionally re-crystallized. The characterisation of the working standard has been presented in paragraph 3.2.S.3.1.1 Spectral analysis. The certificate of analysis is included in the following page.
Reference standard
E - 2/3
Reference standard
E - 3/3
3.2.S.6 CONTAINER CLOSURE SYSTEM
Section F
3.2.S.6 CONTAINER CLOSURE SYSTEM 3.2.S.6.1 Specifications 3.2.S.6.2 Labeling
Container closure system - 1/45
3.2.S.6 - CONTAINER CLOSURE SYSTEM

All the final APIs are packed into plastic drums with double polyethylene bags inside. Each bag is closed with a plastic lace 25 cm long with our trademark, not removable without tampering. The top lid of the drum is sealed with a yellow plastic seal (red for Benzodiazepines) with our trademark, not removable without breaking. We use plastic drums of different capacities, as reported in the following table:
Capacity 6 (*) 8 17(*) 28 50 60 110 125 150
U.M. liters liters liters liters liters liters liters liters liters
The drums are stored in a proper room protected from contamination and polyethylene bags are stored in well-closed containers into sealed packets containing about 100-200 bags. Every plastic drum and polyethylene bag is checked for cleanliness by the operator before use. Every lot of polyethylene bags is checked by Quality Control Laboratory for identity (IR/GC method). The drums, containing the final API and coming from the finishing department, are weighed in a proper room protected from contamination, opened for visual inspection and the labels are replaced with the official ones (one internal on the polyethylene bag and one external on the drums body). PLASTIC DRUMS Characteristics: Body Lid Gasket Closure ring HDPE HMW HDPE Natural gum / Polyurethanic bi-component gum in zinc steel, with sealing lever
(*) The plastic drums of 6 and 17 liters are closed with a screw top lid and sealed with a plastic lace with our trademark non removable without tampering.
POLYETHYLENE BAGS Characteristics: The bags used are made of polyethylene of low density (LDPE) and are food-grade.
CLOSURE SYSTEMS Characteristics: The plastic seals for closing bags are plastic laces about 25 cm long with our trademark, not removable without tampering. The top lid for plastic drums is sealed with a yellow plastic lace (red for Benzodiazepines) with our trademark in order to better avoid tampering.
3.2.S.6.1 Specifications
The specifications of the commercial packaging system are enclosed in the following pages.
Plastic drums (HDPE)
Polyethylene bags
Polymer components: ExxonMobil LD150AC / LD150BW
Polyethylene bag Primary packaging Analytical specifications
IR Polyethylene bag Primary packaging
Antistatic additives
Antistatic additives: Polybatch VLA66 or CESA-Stat 3101
Anti-tampering seals
3.2.S.6.2 Labeling
Label used for the European market
Label used for the USA market
3.2.S.7 - STABILITY
Section G
3.2.S.7 STABILITY 3.2.S.7.1 Stability summary and conclusions 3.2.S.7.1.1 3.2.S.7.1.2 3.2.S.7.1.3 3.2.S.7.1.4 General stability protocol Summary of long term stability Intermediate conditions studies Summary of accelerated stability studies Summary of stress stability studies
3.2.S.7.2 3.2.S.7.3
Post approval stability protocol and stability commitment Stability data 3.2.S.7.3.1 3.2.S.7.3.2 3.2.S.7.3.3 3.2.S.7.3.4 Stability specifications Long term stability Intermediate conditions studies Accelerated stability studies Stress stability studies
Attachments list of Section G Attachment 4 Stability specifications and Stability data
Stability G - 1/115
3.2.S.7.1 STABILITY SUMMARY AND CONCLUSIONS

3.2.S.7.1.1 General stability protocol During the stability study the product is maintained in defined conditions of temperature and humidity and analyzed periodically to establish if changes in the physical and chemical characteristics occur. Stability studies are conducted in compliance with the ICH Guidelines Stability testing of new drug substances and products and GMP for Active Pharmaceutical Ingredients and are classified in two groups:
Temperature Relative humidity Minimum period
40C 2C 75% 5% 6 months
LONG TERM STABILITY INTERMEDIATE CONDITIONS(LTSI)
Temperature Relative humidity Minimum period Maximum period
30C 2C 75% 5% 12 months 5 years
1 Storage of stability samples The product is maintained in a similar packaging of the finished product. This means that the samples are stored in the same type of polyethylene bags and closed with laces. The plastic bags are then packaged into small plastic drums. 2 Lots to put under stability For new products, the first three lots manufactured are put under Accelerated stability and Long Term stability. Every year, one batch is added to the long term stability study, unless no production is done. 3 Retest and expiration dates The Accelerated stability study is conducted for 6 months, but can be extended if requested. The Long term stability study is conducted for at least 5 years. If after this time the product is stable, the expiry date can be fixed in 5 years. On the contrary, if the product is not stable the expiry date depends on the degradation trend of the product.
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4 Frequency of analysis For Accelerated stability (AS), the frequency of testing is the following: For all the lots under testing 0, 1, 2, 3, 6 months
For Long term stability Intermediate Conditions following: For the first 3 batches
(LTSI), the frequency of testing is the
0, 3, 6, 9, 12, 18 months, 2, 3, 4, 5 years
if degradation doesnt occur after 12 months For the additional batches 0, 1, 2, 3, 4, 5 years
Remark: according to the ICH Guidelines Stability testing of new drug substances and products and GMP for Active Pharmaceutical Ingredients, starting from 2000 we have changed the sampling frequency for the additional batches, as reported in the relevant table. Previously, the frequency was the same as for the first 3 batches.
Stability G - 3/115
3.2.S.7.1.2 Summary of long term stability studies The first three validation batches manufactured by Cambrex Profarmaco Milano have been put under long term stability study. For the Long term stability data see paragraph 3.2.S.7.3.2.
3.2.S.7.1.3 Summary of accelerated stability studies The first three validation batches manufactured by Cambrex Profarmaco Milano have been put under accelerated term stability study. For the Accelerated stability data see paragraph 3.2.S.7.3.3.
3.2.S.7.1.4 Summary of stress stability studies During the stress stability study of Emtricitabine, it was possible to identify other degradation products under basic and oxidative conditions. These degradation products were monitored using the three methods cited above and studied with LCMS. According to these studies it became evident that the method for the determination of the related substances is able to separate the isomers of Sulfoxide which are called as isomer 1 and isomer 2, successively. These impurities form principally under the oxidative condition. In addition to this a formation of another impurity was observed under basic condition which was identified as Emtricitabine Hydroxy Analogue, shortly called as Hydroxycytosine. Emtricitabine can be considered stable under solar light after exposure for 10 days and at 105C for 71 hours. Emtricitabine is slightly unstable in aqueous solution as is, in this condition the amount of Fluorouracil increased within the exposure time. Moreover Emtricitabine is unstable under acidic condition for 96 hours at 50C, the main degradation products are Fluorouracil and 5-Fluorocytosine, it is also unstable under basic condition for 96 hours at 50C, the major degradation products are Fluorouracil, an unknown impurity RRT 0.86 assigned as Hydroxycytosine thanks to LC-MS analysis, Sulfoxide Isomers 1 and 2 and 5-Fluorocytosine. The Emtricitabine is very unstable under oxidizing condition, after 48 hours at room temperature the assay has been already decreased to 43.77% and Sulfoxide 1 and 2 were formed in large amount (20.57%, 43.19%).
Stability G - 4/115
Considering the obtained results after the forced degradation study of Emtricitabine, it is demonstrated that the method Emtricitabine_Imp_ 001.001 is able to detect the impurities and it is stability indicating. Moreover as verified in the sequence 120920-P-194-H9 Met USP campioni stressati, comparing the impurity profile of stressed oxide sample applying the USP pending method and the Emtricitabine_Imp_ 001.001, it is evident that the CPM method can resolve the sulfoxide isomers assigned as 1 and 2, and also for the basic sample the critical resolution between Lamivudine and Hydroxycytosine (USP pending method) is improved applying the CPM method. As can be observed from the table 10, the Emtricitabine can be considered stable enantiomerically under all the stressed tested condition, only under basic condition after 96 hours at 50C Emtricitabine 1st 5-Epimer has been detected in a very low amount (0.013%).
Stability G - 5/115
3.2.S.7.2 POST APPROVAL STABILITY PROTOCOL AND STABILITY COMMITMENT
Cambrex Profarmaco Milano, as the manufacturer of Emtricitabine, is committed to perform stability tests not only as pre-approval studies, but also during the complete life of the product. According to our internal procedures, based on the current ICH guidelines: The first three batches of the active ingredients manufactured are put under Accelerated stability and Long Term stability Intermediate Conditions (30C / 75% RH). Every year, one batch is added to the long term stability study, unless no production is done. The first batches, usually 3, manufactured after a major change (submitted to authorities and customers) are put under Accelerated stability and Long Term stability Intermediate Conditions (30C / 75% RH).
The general stability protocol, applied also to this product, is reported in paragraph 3.2.S.7.1.1. Cambrex Profarmaco Milano is committed to withdraw from the market, batches that do not meet specifications and if it is a single occurrence to discuss the issue with proper parties.
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3.2.S.7.3 STABILITY DATA
3.2.S.7.3.1 Stability specifications
Emtricitabine is analyzed according to the Stability specifications enclosed in Attachment 4. The Test methods reported in the stability specification refer to those used for the release of the final active ingredient and resulted to be stability indicating.
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3.2.S.7.3.2 Long Term stability Intermediate Conditions studies The first three validation batches manufactured by Cambrex Profarmaco Milano have been put under Long Term stability Intermediate Conditions (30C / 75% RH) study.
For the Stability data see Attachment 4.
3.2.S.7.3.3 Accelerated stability studies
The first three validation batches manufactured by Cambrex Profarmaco Milano have been put under accelerated term stability study. For the Accelerated stability data see Attachment 4.
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3.2.S.7.3.4 Stress stability studies
Stability G - 9/115
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ATTACHMENT N. 1
SPECIFICATIONS
EMTRICITABINE TS 001.001
Attachment 1 - 1/2
Attachment 1 - 2/2
ATTACHMENT N. 2
ANALYTICAL PROCEDURES
EMBICITRABINE TS 001.001
Attachment 2 - 1/27
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ATTACHMENT N. 4
STABILITY SPECIFICATIONS AND STABILITY DATA
STABILITY SPECIFICATIONS
STABILITY DATA Long term stability data Accelerated stability data
Attachment 4 - 1/10
STABILITY SPECIFICATIONS
Attachment 4 - 2/10
Long term stability data Long term stability studies of EMTRICITABINE are on-going on the following batches:

LONG TERM STABILITY (LTS)
Attachment 4 - 3/10
Attachment 4 - 4/10
Attachment 4 - 5/10
Attachment 4 - 6/10
Accelerated stability data
Accelerated stability studies of EMTRICITABINE are on-going on the following batches:


Attachment 4 - 7/10
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ATTACHMENT N. 5
PRC-TICINUM LAB
Attachment 5 - 1/3
STABILITY TESTING PERFORMED BY PRC-TICINUM LAB
In May 2001, Cambrex Profarmaco Milano, in joint venture with another supplier of active pharmaceutical ingredients, decided to open a new laboratory named PRC TICINUM LAB, located in the Scientific and Technological Pole of Novara, Italy. The aim of this laboratory is to support Cambrex Profarmaco Milano in some analytical and research studies, with the exclusion of the approval and release of raw materials, intermediates and final active ingredients. In particular PRC TICINUM LAB is equipped to store and analyze stability samples. PRC TICINUM LAB is in fact equipped to store the samples into stability chambers automatically regulated and set to comply with the ICH suggested conditions (25C 2 and 60%5 RH), the same applied in Cambrex Profarmaco Milano. PRC TICINUM LAB is committed to follow our stability protocols and use the test methods reported in the current dossier. The new test facility has the capability to perform the intended testing and shows a satisfactory cGMP status. PRC TICINUM LAB has been fully validated by Cambrex Profarmaco Milano through inspection and permits to audit its facility for compliance to cGMP. The analytical methods used to perform the long term stability studies are those described in the current dossier and have been transferred to PRC TICINUM LAB according to a specific procedure (PRC 01/13/PF, Analytical technology transfer). Cambrex Profarmaco Milano is responsible for sending the samples packaged as defined and for handling and presenting the analytical data obtained from PRC TICINUM LAB. PRC TICINUM LAB is responsible for keeping the samples stored in accordance with the established stability protocol and for performing the tests at the agreed timing, by using only the analytical methods that have previously undergone the above mentioned methodology transfer procedure. PRC TICINUM LAB is not allowed to make any change to the official tests methods unless Profarmaco is informed and approves the changes.
Attachment 5 - 2/3
The address of the new laboratory is: PRC (Pharma Research Centre) TICINUM LAB Via Bovio, 6 28100 NOVARA, Italy The same building houses also the Chemical Department for Food, Pharmaceutical and Pharmacological Science of the Piemonte Orientale University. PRC TICINUM LAB is directed by: Dr. Luisa Zangirolami University degree in Pharmaceutical Chemistry and Technology University of Padua Dr Luisa Zangirolami has a very broad experience in Pharmaceutical Chemistry having covered, during previous professional experiences, the role of Quality Control and Regulatory Affair Director in pharmaceutical and API manufacturer companies, many of them FDA approved. All personnel of PRC TICINUM LAB owns adequate knowledge, experience and training to perform analytical tests, as requested by the cGMP. PRC TICINUM LAB is equipped with analytical instrumentation like Hplc, GC, IR, UV, DSC etc. (mostly supplied from Agilent) subjected to all the qualification and calibration procedures, as requested by the cGMP. PRC TICINUM LAB has developed, in cooperation with Cambrex Profarmaco Milano, an internal Quality System based on official standard operating procedures that are strictly followed by all analysts and supervisors. Cambrex Profarmaco Milano can therefore guarantee that all the activities performed by PRC TICINUM LAB are in compliance with the cGMP and the relevant ICH or FDA guidelines.
Attachment 5 - 3/3
Cambr rexProfarmacoMilanoS.r.l. .
Emtricitabine, 10 0/2012 Ed 01.00/2012-ASMF
Att tachment 6 - 1/68
Emtric citabine, 10/2012 Ed 01.00/2012-A ASMF
Atta achment 6 - 10/68
Emtric citabine, 10/2012 Ed 01.00/2012-A ASMF Atta achment 6 - 19/68
Emtric citabine, 10/2012 Ed 01.00/2012-A ASMF Atta achment 6 - 33/68
Emtricitabine, 10/ /2012 Ed 01 1.00/2012-A ASMF
Emtricitabine, 10/ /2012 Ed 01 1.00/2012-A ASMF Atta achment 7 - 20/126
Emtricit tabine, 10/2 2012 Ed 01.00/2012-AS SMF
Attac chment 7 - 100/126 1
Emtricit tabine, 10/2 2012 Ed 01.00/2012-AS SMF Attac chment 7 - 126/126 1
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Emtric citabine, 10 0/2012 Ed 01.00/2012-A ASMF
Emtric citabine, 10 0/2012 Ed 01.00/2012-A ASMF Attac chment 8 - 46/165 4
Emtric citabine, 10/ /2012 Ed 01 1.00/2012-A ASMF
Attach hment 8 - 100/165

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