ASMF 01-00-2012- AP

Cambrex Profarmaco Milano S.r.l.

EMTRICITABINE

ACTIVE SUBSTANCE MASTER FILE

Ed. 01.00/2012-ASMF

NEW SUBMISSION

OCTOBER 2012


Emtricitabine, 10/2012 Ed 01.00/2012-ASMF Index - 1/3

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

MODULE 1 - ADMINISTRATIVE INFORMATION AND PRESCRIBING INFORMATION

1.4.1 Information about the expert quality Annex I Environmental risk assessment

MODULE 2 - COMMON TECHNICAL DOCUMENT SUMMARIES

2.3 Quality overall summary (AP) 2.3 Quality overall summary (RP)*

MODULE 3 - QUALITY 3.2.S DRUG SUBSTANCE SECTION A 3.2.S.1 GENERAL INFORMATION A 1/3

3.2.S.1.1 Nomenclature A - 2

3.2.S.1.2 Structure A - 2 Structural formula A - 2 Molecular formula A - 2 Molecular weight A - 2

3.2.S.1.3 General properties A - 3 SECTION B1 3.2.S.2 MANUFACTURE B1 1/30 3.2.S.2.1 Manufacturer B1 - 2 3.2.S.2.2 Description of manufacturing process and

process controls B1 - 3 3.2.S.2.2.1 Scheme of the synthesis B1 - 3

3.2.S.2.2.2 Scheme of the manufacturing process* B1 - 4 3.2.S.2.2.3 Brief description of the synthesis* B1 5 3.2.S.2.2.4 Flow-sheet* B1 6 3.2.S.2.2.5 Detailed description of the manufacturing process* B1 - 8 3.2.S.2.2.6 Reprocessing* B1 - 20 3.2.S.2.2.7 In-process controls* B1 - 21

SECTION B2 3.2.S.2.3 Control of materials* B2 1/17

3.2.S.2.3.1 Raw materials list* B2 - 2 3.2.S.2.3.2 Raw materials sampling procedure* B2 - 3

3.2.S.2.3.3 Raw materials specifications and test methods* B2 - 3

3.2.S.2.4 Control of critical steps and intermediates* B2 - 5 3.2.S.2.5 Process validation* B2 - 16 3.2.S.2.6 Manufacturing process development* B2 - 17



SECTION C 3.2.S.3 CHARACTERIZATION C 1/73

3.2.S.3.1 Elucidation of structure and other characteristics C - 2 3.2.S.3.1.1 Spectral analysis C - 2

- NMR Spectrum C - 2 - IR Spectrum C - 12

- Mass Spectrum C - 16 - Elemental analysis C - 25

3.2.S.3.1.2 Potential isomerism C - 28 3.2.S.3.1.3 Polymorphism C - 30

3.2.S.3.2 Impurities C - 43 3.2.S.3.2.1 Related substances and degradation

products C - 43 3.2.S.3.2.2 Description of impurities C - 52 3.2.S.3.2.3 Characterization of impurities C - 55

SECTION D 3.2.S.4 CONTROL OF DRUG SUBSTANCE D 1/7

3.2.S.4.1 Specification D - 2 3.2.S.4.2 Analytical procedures D - 2 3.2.S.4.3 Validation of analytical procedures D - 2 3.2.S.4.4 Batch analysis D - 3 3.2.S.4.5 Justification of specification D - 7 SECTION E 3.2.S.5 REFERENCE STANDARD E 1/3 SECTION F 3.2.S.6 CONTAINER CLOSURE SYSTEM F 1/45

3.2.S.6.1 Specifications F - 3 3.2.S.6.2 Labeling F - 45

SECTION G 3.2.S.7 STABILITY G 1/115

3.2.S.7.1 Stability summary and conclusions G - 2 3.2.S.7.1.1 General stability protocol G - 2

3.2.S.7.1.2 Summary of long term stability studies G - 4 3.2.S.7.1.3 Summary of accelerated stability studies G - 4 3.2.S.7.1.4 Summary of stress stability studies G - 4

3.2.S.7.2 Post approval stability protocol and stability commitment G - 5

3.2.S.7.3 Stability data G - 6 3.2.S.7.3.1 Stability specifications G - 6 3.2.S.7.3.2 Long term stability studies G - 7 3.2.S.7.3.3 Accelerated stability studies G - 7 3.2.S.7.3.4 Stress stability studies G - 9



ATTACHMENTS

Attachment 1 Section D Specifications Attachment 2 Section D Analytical procedures Attachment 3 Section B2 Raw material specifications* Attachment 4 Section G Stability specifications and Stability data Attachment 5 Section D Copper determination by atomic absorption - validation Attachment 6 Section D Validation report for assay by HPLC Attachment 7 Section D Enantiomeric purity and chiral identity by hplc-Validation Attachment 8 Section D Related substances by HPLC Validation Attachment 9 Section B2 Starting material qualification*

*) Restricted part


Emtricitabine, 10/2012 Ed 01.00/2012-ASMF Annex I - Environmental risk assessment - 1/3

Module 1 - Annex I

ENVIRONMENTAL RISK ASSESSMENT



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

S

O N

NO

F

NH2

OH

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 (health and safety) D.L. 334 of 17/8/1999 (risk activities) A.I.A./IPPC of 7/9/2007 (environmental integrated authorization of the site) D.P.R. 203 of 24/5/1988 (air) D.L. 152 of 11/5/1999 (water) D.L. 22 of 5/2/1997 (wastes) 94/67CE of 16/12/1994 (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.


Emtricitabine, 10/2012 Ed 01.00/2012-ASMF Module 1 - Information about the expert - 1/3

MODULE 1

ADMINISTRATIVE INFORMATION AND

PRESCRIBING INFORMATION

1.4.1 - INFORMATION ABOUT THE EXPERT QUALITY

ANNEX I - ENVIRONMENTAL RISK ASSESSMENT



1.4.1 INFORMATION ABOUT THE EXPERT - QUALITY Name of the expert: Dr. Fulvio Leone Piselli

Address: 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 Milan, November 9 1933 Address Via Sangro 11 , Milan ITALY Employed 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


Emtricitabine, 10/2012 Ed 01.00/2012-ASMF APPLICANT PART Module 2 QOS - 1/12

MODULE 2

COMMON TECHNICAL DOCUMENT SUMMARIES

2.3 - QUALITY OVERALL SUMMARY

APPLICANT PART



2.3 QUALITY OVERALL SUMMARY APPLICANT PART

2.3.S DRUG SUBSTANCE 2.3.S.1 GENERAL INFORMATION

EMTRICITABINE

S

O N

NO

F

NH2

OH

Molecular formula: C8H10FN3O3S Molecular weight: 247.3 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 143491-57-0

Cambrex Profarmaco Milano code 194



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)



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



Scheme of the synthesis of EMTRICITABINE (paragraph 3.2.S.2.2.1)

O

O

S

OOH

O

O

S

OCl

O

O

S

O N

NO

F

NH2

NH

NO

F

NH2

SOCl2

HYDOXA

S

O N

NO

F

NH2

OH

S

O N

NO

F

NH2

OH

N

NO

F

NH

Si Si

Hexamethyldisilazan

Diisopropylethylamine

Chloro solution

1) NaBH42) Maleic Acid

EMTRICITABINE (FTC BASE)

FTC Menthylester

FLUOCY

Silylcytosine solution

FTC maleate

XX Wet isopropanol

Triethylamine Isopropanol

(Maleate salt 1:1)

+

CRYSTALLIZED EMTRICITABINE(FTC BASE)



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 cis-2R,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.



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-oxathiolane-

2-carboxylate (MENTHYL ESTER)



2.3.S.4 CONTROL OF THE DRUG SUBSTANCE Specification for EMTRICITABINE (paragraph 3.2.S.4.1):



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.



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.



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.



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 Batch size (Kg) Date of manufacture 010102 25 Kg 12/2012 010304 32 Kg 12/2012 010506 34 Kg 12/2012

These lots have been stored under the following conditions: LONG TERM STABILITY (LTS)

Temperature 30C 2C Relative humidity 75% 5% Accelerated stability studies (paragraph 3.2.S.7.3.3) Accelerated stability studies of EMTRICITABINE are on-going on the following batches:

Batches Batch size (Kg) Date of manufacture 010102 25 Kg 12/2012 010304 32 Kg 12/2012 010506 34 Kg 12/2012

These lots have been stored under the following conditions: ACCELERATED STABILITY (AS)

Temperature 40C 2C Relative humidity 75% 5%


Emtricitabine, 10/2012 Ed 01.00/2012-ASMF Module 3

MODULE 3 - QUALITY

3.2.S - DRUG SUBSTANCE


Emtricitabine, 10/2012 Ed 01.00/2012-ASMF General information A - 1/3

3.2.S.1 GENERAL INFORMATION

Section A

3.2.S.1 GENERAL INFORMATION 3.2.S.1.1 Nomenclature 3.2.S.1.2 Structure

- Structural formula - Molecular formula - Molecular weight

3.2.S.1.3 General properties



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 143491-57-0

Cambrex Profarmaco Milano code 194

3.2.S.1.2 - STRUCTURE

Structural formula

S

O N

NO

F

NH2

OH

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)


Emtricitabine, 10/2012 Ed 01.00/2012-ASMF Manufacture B1 - 1/30

3.2.S.2 - MANUFACTURE

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*

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*

Attachments list of Section B2

Attachment 3 Raw materials specifications* Attachment 9 Starting material qualification*

* Restricted part



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

OOH

O

O

S

OCl

O

O

S

O N

NO

F

NH2

NH

NO

F

NH2

SOCl2

HYDOXA

S

O N

NO

F

NH2

OH

S

O N

NO

F

NH2

OH

N

NO

F

NH

Si Si

Hexamethyldisilazan

Diisopropylethylamine

Chloro solution

1) NaBH42) Maleic Acid

EMTRICITABINE (FTC BASE)

FTC Menthylester

FLUOCY

Silylcytosine solution

FTC maleate

XX Wet isopropanol

Triethylamine Isopropanol

(Maleate salt 1:1)

+

CRYSTALLIZED EMTRICITABINE(FTC BASE)


Emtricitabine, 10/2012 Ed 01.00/2012-ASMF Characterization C - 1/73

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



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 .



IR SPECTRUM

The FT-IR spectrum of Emtricitabine WS was recorded on a JASCO FT-IR 420 Spectrometer.



MS SPECTRUM

The MS analysis of Emtricitabine WS was performed using an Agilent 6310. The fragmentation peaks conform to the proposed structure.



ELEMENTAL ANALYSIS

The results of the elemental analysis of Emtricitabine WS agree with the proposed structure, as reported in the following table.



3.2.S.3.1.2 Potential Isomerism The molecule has two stereocenters indicated in the picture below. The isomer of interest is the cis-2R,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



3.2.S.3.1.3 Polymorphism



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-oxathiolane-

2-carboxylate (MENTHYL ESTER)

The standard HPLC purity profile for Emtricitabine manufactured by Cambrex Profarmaco Milano is the following:



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.



Metal catalysts

In the process are not used metal catalysts.



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

NH

NO 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

OH

S

O N

NO

F

NH2

O

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.



4) 5-Fluoro-1-[(2R,3RS,5S)-2-(hydroxymethyl)-3-oxo-1,3-oxathiolan-5-yl] cytosine EMTRICITABINE SULFOXIDES

OH

S+

O N

NO

F

NH2

O

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

OH

S

O N

NO 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).



6) 5-Fluoro-1-[(2R,5S)-2-(hydroxymethyl)-1,3-oxathiolan-5-yl]uracil

FLUOROURACIL ANALOG

OH

S

O N

NH

O

F

O

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-2-

carboxylate MENTHYL ESTER

O

S

O N

NO

F

NH2

O

This substance is an intermediate of the process, isolated in the synthesis of Emtricitabine. Its carry over appeared to be very low.



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.



Emtricitabine Acid

OH

S

O N

NO

F

NH2

O

The elucidation of the chemical structure of the impurity Emtricitabine acid is based on 1H-NMR.



L-Menthyl 5-(2R,5S)-[4-amino-5-fluoro-2-oxopyrimidin-1(2H)-yl]-1,3-oxathiolane-2-carboxylate ( MENTHYL ESTER)

O

S

O N

NO

F

NH2

O

The elucidation of the chemical structure of the impurity Menthyl ester is based on 1H-NMR,



cis-butenedioic acid MALEIC ACID



4-Amino-5-fluoropyrimidin-2-(1H)-one

(FLUOROCYTOSINE)



(2)(-)-1-[(2R,5S)-2-(Hydroxymethyl)-1,3-oxathiolan-5-yl]cytosine LAMIVUDINE

OH

S

O N

NO NH2


Emtricitabine, 10/2012 Ed 01.00/2012-ASMF Control of drug substance D - 1/7

3.2.S.4 - CONTROL OF DRUG SUBSTANCE

Section D

3.2.S.4 CONTROL OF DRUG SUBSTANCE

3.2.S.4.1 Specification 3.2.S.4.2 Analytical procedures 3.2.S.4.3 Validation of analytical procedures 3.2.S.4.4 Batch analysis 3.2.S.4.5 Justification of specification

Attachments list of Section D

Attachment 1 Specifications Attachment 2 Analytical procedures Attachment 6 Assay by HPLC - validation

Attachment 7 Enantiomeric purity and chiral identity by hplc Attachment 8 Related substances by HPLC Validation



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 Attachment 6 Enantiomeric purity and chiral identity by hplc Validation Attachment 7 Related substances by HPLC Validation Attachment 8



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 010102 010304 010506

Manufacturing date 12/2012 12/2012 12/2012

Batch size 25 Kg 32 Kg 34 Kg



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.


Emtricitabine, 10/2012 Ed 01.00/2012-ASMF Reference standard E - 1/3

3.2.S.5 - REFERENCE STANDARD

Section E



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.


Emtricitabine, 10/2012 Ed 01.00/2012-ASMF Container closure system - 1/45

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



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 U.M.6 (*) liters 8 liters 17(*) liters 28 liters 50 liters 60 liters 110 liters 125 liters 150 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 HDPE HMW Lid HDPE Gasket Natural gum / Polyurethanic bi-component gum Closure ring 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


Emtricitabine, 10/2012 Ed 01.00/2012-ASMF Stability G - 1/115

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 General stability protocol 3.2.S.7.1.2 Summary of long term stability Intermediate conditions 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 Intermediate conditions studies 3.2.S.7.3.3 Accelerated stability studies 3.2.S.7.3.4 Stress stability studies

Attachments list of Section G

Attachment 4 Stability specifications and Stability data



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:

ACCELERATED STABILITY (AS)

Temperature 40C 2C Relative humidity 75% 5% Minimum period 6 months

LONG TERM STABILITY INTERMEDIATE CONDITIONS(LTSI)

Temperature 30C 2C Relative humidity 75% 5% Minimum period 12 months Maximum period 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.



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 (LTSI), the frequency of testing is the following:

For the first 3 batches 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.



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%).



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%).



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.



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.



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.



3.2.S.7.3.4 Stress stability studies


Emtricitabine, 10/2012 Ed 01.00/2012-ASMF Attachment 1 - 1/2

ATTACHMENT N. 1

ASMF 01-00-2012- AP

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