Unassociated Document



  
















Cobra Biomanufacturing Plc

The
Science Park,

Keele,

U.K.

ST5
5SP











 





















   

   Executive
Summary




Advaxis
has developed a recombinant attenuated L.
monocytogenes for
vaccination against HPV E7 expressing tumors. Advaxis will require the
production of approximately [ * ] cfu of
L.
monocytogenes non-cGMP
preclinical material and [ * ] cfu for
clinical trial use produced following cGMP guidelines. The program will involve
the following phases:

Phase
I

	·  	
      Transfer
      of current Listeria
      culture and analysis methods 



	·  	
      Two
      month feasibility study and process development
program



	·  	
      Animal
      component free growth media recommendation



	·  	
      Analytical
      methods development and host characterization methods




Phase
II

	·  	
      cGMP
      Master Cell Bank production



	·  	
      Toxicology
      material



	·  	
      Manufacture
      of clinical material



	·  	
      Development
      of product stability tests 



	·  	
      Quality
      assurance review 



	·  	
      Bulk
      product release for fill/finish






To
achieve these goals, Advaxis will require the collaboration of a partner
with:

	·  	
      Specialized
      facilities for plasmid DNA manufacture



	·  	
      Experience
      in plasmid DNA manufacture according to cGMP 



	·  	
      Successful
      track record producing material for clinical trials in the
    USA



	·  	
      Experience
      in meeting regulatory requirements in facilities and
      documentation






Cobra
has both the expertise and the facilities available to meet the project
deliverables required by Advaxis in the timescales attached.




Cobra
Biomanufacturing Plc

Cobra is
a full
service, world class Contract Manufacturing Organization that manufactures and
supplies DNA-based therapeutics for the pharmaceutical and biotechnology
industries. Cobra provides contract services spanning pre-clinical to early
Phase III scale production and supply of biological products. These services
include the cGMP manufacture of DNA, recombinant protein, viruses, mammalian
cell products and cell banking.  




Currently,
four clinical trials in the USA are being conducted using products manufactured
at Cobra. Additionally, there are clinical trials in Europe, Africa, China, and
Australia using products manufactured at Cobra.




Cobra has
a Type II Drug Master File (DMF) lodged with the FDA covering DNA manufacture.
We were last inspected by the MCA in August 2002 and found to be cGMP compliant.
Cobra provides a comprehensive analytical and documentation package for
regulatory filing.

 


 


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Introduction




Cobra
Biomanufacturing
is a full service, world class Contract Manufacturing Organization that
manufactures and supplies DNA-based therapeutics for the pharmaceutical and
biotechnology industries. Cobra provides contract services spanning pre-clinical
to early Phase III scale production and supply of biological products. These
services include the cGMP manufacture of DNA, recombinant protein, viruses,
mammalian cell products and cell banking. The company also undertakes process
development programs for recombinant protein and gene therapy products.





Company
History




Cobra was
founded in 1992 as a start up biotechnology company specializing in gene therapy
and has been operating from facilities at the Keele Science Park for the past 8
years. The manufacturing division was originally established in order to
expedite Cobra Therapeutics own R & D programs. Investments were made in
cGMP manufacturing facilities and the development of technology for scaleable
manufacture of DNA and protein based pharmaceuticals. In 1998 the manufacturing
division began to offer cGMP manufacturing services to the pharmaceutical
industry. Cobra Therapeutics became a wholly owned subsidiary of ML Laboratories
in 2000. 




In June
2002, following a successful IPO on London’s AIM, Cobra Biomanufacturing was
established as an independent company
with an exclusive focus on custom manufacturing of bio
therapeutics.




Cobra’s
corporate objective is to continue to grow as a major contract supplier of DNA,
virus and protein based therapeutics for clinical trials and of licensed
biopharmaceutical products for commercial sale.




Manufacture
of DNA Therapeutics for Clinical Trials




Cobra has
established a worldwide reputation in the manufacture of plasmid DNA
therapeutics and is supporting clinical trials in the USA, Europe, Africa,
China, and Australia.




Every
project undergoes a technology transfer of your existing expression system to
utilize our scale-up expertise before initiating the cGMP manufacturing program.
During the evaluation stage genetic stability and relative productivity will be
determined in shake flask experiments. The fermenter productivity of the
transformed host strain will then be evaluated in a scale down (5L)
evaluation.
This
initial optimisation is to achieve the maximum productivity in the fermentation,
ensure yield and product purity throughout the purification process. It is
essential for the identification
of potential difficulties with your plasmid. The process development work is
necessary because in our experience 





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there can
often be a 5-10 fold difference in productivity between strains (even with
similar plasmid backbones) and genetic instability is observed with some
plasmids. By assessing the plasmid at the beginning of the program, we can
accurately estimate the yields of clinical material that you will
obtain.

 

Cobra has
substantial experience working with Kanamycin and Tetracycline resistant
plasmids and has also developed and been granted patents covering an
antibiotic-free plasmid DNA manufacturing process, the Operator Repressor
Titration (ORT) System. 

A Type II
DMF (Drug Master File) has been lodged with the FDA covering DNA
manufacture.




Facilities




Cobra has
over 11,000 square feet of space used for a process development facility,
separate QA/QC laboratories, and dedicated cGMP manufacturing facilities. The
existing cGMP manufacturing facility includes 4,500 square feet of EU Grade C
Clean Room space required for key stages in the manufacture of biopharmaceutical
products. The cGMP facility has two microbial production suites. The [ * ]
fermenter suite ([ * ] working volume) is for Phase I and II clinical trial
material. There is a [ * ] fermenter ([ * ] working volume) that is used to
provide an inoculum for the larger fermenter. The [ * ]fermenter suite ([ * ]
working volume) is for Phase II and III clinical trial material. 




Additionally,
there are two virus production suites with [ * ] and [ * ] fermenters ([ * ] and
[ * ] working volume) utilising adenovirus and baculovirus expression systems
for manufacture of Phase I and II clinical trial material.




Quality




Cobra is
committed to conducting its manufacturing activities in accordance with
appropriate current Good Manufacturing Practice (cGMP) and Good Control
Laboratory Practice (GLP) regulations and/or guidelines. The latest inspection
from the UK Medicines Control Agency (MCA) was in August 2002 with a compliance
statement received several weeks later. Cobra’s QA group ensures that the
products manufactured by the division meet appropriate standards of safety,
quality, and efficacy. The QA group oversees manufacture at all stages and is
responsible for testing, release, storage, and arranges shipment of the drug
product. Overseas shipping and safe passage 

through
Customs is easily co-ordinated and sub-contracted to BioCair, Inc or World
Courier.




Key
Personnel 




Cobra’s
belief that quality individuals result in quality products is reflected in the
key personnel that will be involved in the manufacture of Advaxis’ L.
monocytogenes for
clinical trials.





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    Julian
      Hanak B.Sc.
      (Hons), MSc., Director
      of Production
      

  




After
gaining an honours degree in Biochemistry at University College London, Julian
obtained an MSc at the University College of North Wales and then trained in
cell culture and microbial fermentation at the National Institute of Medical
Research. He then moved to the Bioproducts Laboratory (Elstree) where his duties
involved the pilot scale production of human monoclonal antibodies for clinical
trials. He was also responsible for running a sterile fill operation and
supervising the commissioning of a new cGMP production suite.




In 1992,
Julian moved to Zeneca Pharmaceuticals where he was involved with the process
development of several immunotherapy products and the development of virus
expression systems for protein production. He joined Cobra in 1994 and took over
responsibility for production in 1995.




Geoff
Sharpe BSc.,
PhD., CChem, MRSC.,
Director
of Quality
Assurance 




Geoff has
over 25 years experience in pharmaceutical biotechnology with over 12 years
experience in Quality Assurance. After having gained a degree in Applied
Chemistry at Liverpool, Geoff trained as a research chemist working for ICI
Corporate Laboratory. He later worked at the ICI Corporate Bioscience Group and
went on to complete a PhD in molecular biology at Leicester University.





In 1991
he transferred to ICI Pharmaceuticals (now AstraZeneca) where he was involved
with the cloning and expression of recombinant proteins and managed the
corporate DNA sequencing laboratory. In 1993, he moved to Zeneca
Pharmaceuticals. In the pharmaceutical department Geoff managed a team involved
in the development, manufacture, and release of both small molecule and
biotechnology based therapeutics. In 1996 he joined Cobra as their Quality
Assurance Manager and has been trained as a Qualified Person under Article 23 of
Directive 75/319/EEC.




Amanda
Weiss BSc.,
MSc.,
Section
Head Fermentation




Amanda
was trained at the University of Birmingham, Centre for Biochemical Engineering
before joining Cobra in 1996 as a fermentation scientist. Amanda has expertise
in microbial and mammalian cell culture, scale-up design and large-scale


manufacture
of biopharmaceuticals. She was also involved with the exemplification and
publication of Cobra’s ORT technology. Amanda has successfully managed the
fermentation aspect of Cobra’s manufacturing operations for over 5
years.




Tony
Hitchcock BSc,
Section
Head Microbial Products




Tony has
over 19 years’ experience in the large-scale manufacture of biopharmaceuticals.
Tony has held positions in the Blood Products Laboratory (Elstree) and at Zeneca
Pharmaceuticals in the protein process development department. Tony was a
founding staff
member of Cobra and has been responsible for the development of much of Cobra’s
DNA manufacturing technology. Tony has published several papers in the field and
is an inventor on two families of Cobra’s process patents.


 



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Roy
Cowell BSc.
(Hons), CChem, MRSC,
Section
Head Quality
Control 




Roy has
16 years’ experience of analytical development and quality control of
pharmaceuticals within the associated regulatory framework. Ten years employed
by Zeneca (now AstraZeneca) Pharmaceuticals working on new chemical entities and
candidate biotherapeutics and six years employed at Cobra working on candidate
DNA products. Roy is currently undergoing training leading to eligibility for
Qualified Person status.




Joy
Manley BSc, Senior
QA Microbiologist




Joy is
currently responsible for developing, validating, and applying suitable testing
regimes that help to assure clean room suitability and equipment cleanliness.
New test methods are designed and validated for plant systems and for cleaning
as required. Both standard and novel microbiological methods are developed and
used to characterize cell banks. She has experience in working with
microorganisms from both pharmaceutical and clinical backgrounds, previously
working for Fisons and The Public Health Laboratory.




David
Thatcher, Chief Executive




David was
trained as a protein chemist at the Universities of Newcastle on Tyne and
Edinburgh. In 1981 he moved to Biogen SA in Geneva where he worked on the
isolation of recombinant cytokines. In 1985 he became Director of Process
Development of Biogen, Inc. in Cambridge, MA, where he was responsible for the
development of large-scale processes for the production of gamma interferon,
GM-CSF and several other products. 




In 1988
he left Biogen and joined Zeneca Pharmaceuticals as head of their Protein
Production Lab where he was responsible for the production of a number of
biopharmaceutical products for clinical evaluation. In 1994 he joined Cobra and
has been responsible for managing the evolution of Cobra’s manufacturing
technology and developing the contract manufacturing business into an
independent company with a successful initial public offering.





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Quotation
O422




Description        Price




Phase
I




[
* ] 







Phase
II




[
* ]







Notes:

Stage
I





	1.	Execution
      of Material Transfer Agreement. Advaxis methods for recombinant
      Listeria
      culture and analysis will be transferred to Cobra. This will include
      plasmid isolation, plasmid and host identity, plasmid and host stability,
      cryopreservation, and protocol for plasmid
isolation.







	 2.	Characterization
      and strain history of the untransformed L.
      monocytogenes
      will be addressed by Advaxis. Advaxis will also be responsible for plasmid
      sequence and/or detailed restriction maps. Host and plasmid information is
      required for the GMO risk assessment. A letter from Dr. Paterson
      addressing the mobility of L.
      monocytogenes is
      requested.








	3.	Advaxis
      will supply [ * ] vials of a transformed research cell bank (mid log
      phase) of L.
      monocytogenes
      with documentation sufficient to make the research bank suitable for
      generation of the cGMP Master Cell Bank.








	4.	A
      two-month feasibility study will be undertaken to determine the growth
      kinetics of Listeria
      (latest
      harvest point) in various growth media. The study will also involve
      bioreactor growth, analysis of log phase, determination of yield, and
      number of cell doublings in vivo before maintenance of virulence is lost.
      We suggest running Stage II at the same time as Stage I to reduce the
      timeline to cGMP manufacture. 






Stage
II





	4.	An
      animal component free growth media will be recommended following
      evaluation
      of the existing media formulations with suitable alternatives. The media
      evaluated will be from published references for media used in Listeria
      culture.








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	5.	Development
      of a cryopreservation media suitable for administration to
    patients.







	6.	Analytical
      methods will be developed to meet FDA regulatory requirements for a
       live
      attenuated bacterial vaccine. Methods developed will include:







	o  	
      host
      identity



	o  	
      plasmid
      identity (restriction mapping or sequencing)



	o  	
      culture
      purity (monosepsis)



	o  	
      viable
      count.







	7.	Host
      characterization methods will be developed for the
following:






	o  	
      phenotype
      auxotrophies and markers



	o  	
      morphology






	o  	
      specific
      media for identification



	o  	
      gram
      strain









	8.	Cobra
      will supply Dr. Paterson with [ * ] of log phase culture for a hemolysin
      assay. Additionally, Cobra will supply Dr. Paterson with three samples of
      [ * ] for a mouse tumor challenge to study maintenance of
    virulence.









	9.	The
      following documentation will be provided:






Technical
Report





	10.	Confirmation
      of price estimates for cGMP manufacture at this point, dependent upon
      successful technology transfer, feasibility study and process development.
      






Stage
III





	
      11.
	
      Cell
      banking will only proceed based upon the feasibility study achieving cell
      densities of at least [ * ] viable
      cells per litre of culture. The Master Cell Bank will be manufactured
      under cGMP in accordance with the latest CPMP guidelines and MCA guidance.
      A Type II Drug Master File has been lodged with the FDA covering these
      procedures. Cells will be cryopreserved in mid log growth at a density of
      between [ * ] to [ * ] cfu/ml. 







Pricing
for the Working Cell Bank is based on production immediately following the
Master Cell Bank. Characterization for the Working Cell Bank is free of charge
if concurrent with Master Cell Bank testing.





	
      12.
	
      A [
      * ]-vial cGMP Master Cell Bank and Working Cell Bank will be released
      according to the agreed program. 









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      13.
	
      The
      Master Cell Bank and Working Cell Bank will be characterised using the
      following range of tests:







	·  	
      Confirmation
      of species (API Listeria)



	·  	
      Confirmation
      of strain by partial genotyping 



	·  	
      Plasmid
      stability by serial sub-culture



	·  	
      Counter
      selection for monosepsis



	·  	
      Plasmid
      identity by restriction digest



	·  	
      DNA
      sequence of the plasmid (to be invoiced
separately).







	14.	The
      non-cGMP material for use in toxicology studies, stability testing, and
      quality control
      lot release will produced at the [ * ] scale with a yield of [ * ]
      based
      upon the feasibility study achieving cell densities of at least [ * ]
      viable cells per litre of culture.







	15.	The
      following documentation will be provided to support a Regulatory
      filing:






Certificates
of Analysis
Analytical
Reports

 

Stage
IV





	16.	cGMP
      Manufacture:
      Prices are estimates without knowledge of the results of the Phase I
      Feasibility Program and may require variances to this proposal. If the
      productivity of the strain cannot be developed to achieve cell densities
      of at least [ * ] viable cells per litre of culture the delivery of [ * ]%
      of final bulk material cannot be guaranteed. The expected quantity of bulk
      and scale required will be advised as soon as it is determined during the
      Phase I Feasibility and Development Program and prior to initiation of the
      Phase II cGMP manufacturing program. If cell densities of [ *
      ] cells
      per litre are obtained in the feasibility study then a [ * ] fermentation
      should yield the requested [ * ] clinical material. If the desired cell
      densities of [ * ] cells per litre are not achieved, then the cGMP
      manufacturing program will be renegotiated.






Stage
V





	17.	Product
      Stability Testing will be required, but will be negotiated as a separate
      contract once the methods have been developed and the protocol agreed by
      Advaxis after FDA discussions. Stability tests for genetic stability; cell
      bank stability and bulk drug stability will be developed once a program is
      agreed upon. The figure provided is for budgetary
purposes.






Stage
VI






	18.	Documentation.







The
following documentation will be provided:






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Certificate
of Analysis

Technical
Summary to support regulatory filing

A copy of
the completed BMR will be provided. 





	19.	Specifications






Cobra
warrants that upon delivery of the Product to Advaxis, Inc. the Product
shall:

	·  	
      Have
      been manufactured in accordance with cGMP. 



	·  	
      Be
      in conformity with the provisional draft specifications as attached to
      this document.






	·  	
      That
      Cobra will provide Product of sufficient quality for human clinical
      use.



	·  	
      In
      the event the Product fails to meet any of the specification described
      above, the final determination as to the suitability of the product for
      human clinical use shall be determined by Advaxis, Inc., who may consult
      with the appropriate offices of the US FDA or other regulatory
      agencies.






Stage
VII





	
      20.
	
      Fill/Finish
      will be subcontracted to BioReliance. A quote cannot be provided until the
      type of container, number of vials and other variables have been
      determined. The figure provided is for budgetary purposes.
  








	20.	The
      costs of consumables have not been included in this quotation and will be
       
      billed directly to the customer without additional charge.








	21.	The
      cost of subcontracted work has not been included in this quotation and
      will be billed
      directly to the customer (plus a [ * ]% handling
charge).







	
      22.
	
      Cobra
      will take responsibility for shipment. The price of shipment of bulk,
      dosage forms and samples and insurance thereof is excluded from this
      contract. Shipping will be arranged in consultation with the customer and
      will be billed directly to the customer (plus a [ * ]% handling
      charge).








	23.	Cobra
      and/or Advaxis, Inc. may wish to issue a press release relating to this
      contract. However, prior to any information being disclosed written
      approval must be obtained from the other party.



 


	24.	The
      Customer agrees to pay reasonable travel expenses connected with Cobra
      staff attending
      meetings, other than those on Company premises and requested by the
      Customer.
      








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      25.
	
      Cobra
      Bio-Manufacturing plc’s O422 Phase I Terms and Conditions and O422 Phase
      II Terms and Conditions apply to this work and acceptance of this
      quotation implies acceptance of these Terms and Conditions.
    







How
to Proceed




Please
return a signed copy of the enclosed contract with your formal Purchase Order to
Cobra Biomanufacturing. 




Cobra
Biomanufacturing Plc

The
Science Park

Keele,
United Kingdom ST5 5SP

Phone:
011 44 1782 714181

Fax: 011
44 1782 714168




When
timing is critical a faxed version is acceptable, but an original must be signed
and returned within fourteen days. Upon receipt Cobra will notify the client of
acceptance within 72 hours.









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Advaxis,
Inc.

212
Carnegie Center, Suite 206

Princeton,
NJ 08540 USA




July 7,
2003




Contract
O422

Determination
of Manufacturing Parameters, Process Development and cGMP Manufacture
of L.
monocytogenes

  

(a)
Phase I: Two-month Feasibility Study and Development
Program




Price 

(Line
items: a + b + c):




Total:       $[
* ]




Terms
of Payment




The
following payment terms will apply: On receipt of a signed copy of the Contract,
Cobra Biomanufacturing Plc. will begin Phase I. Upon commencement of the work
program, Advaxis will be invoiced for $[ * ] net 30 days and will be invoiced
the remaining $[ * ] appropriately on a monthly basis for the length of the
program. The final invoice will be sent before [ * ]. 




This
phase of the program is governed by the Terms and Conditions set out in the
attached document “ O422 Phase I Terms and Conditions”.




Phase
II: Pre-Clinical and GMP Manufacture




Price
Estimate is based on [ * ] cGMP manufacture: 

(Line
items: d + e + f + g + i +k + l + m + n + o)




Total:       $[
* ] (excluding Fill/Finish)




Terms
of Payment




The
following payment terms will apply: On receipt of a signed copy of the Contract,
Cobra Biomanufacturing Plc. will hold a slot for Advaxis without a deposit.
Advaxis will be notified of any request for the slot and may reserve the slot
with a [ * ]% deposit. Receipt of this payment will reserve the production slots
as per the agreed program. On commencement of the work program, [ * ]% of the
cost will be appropriately invoiced on a monthly
basis for the length of the program, with the remaining [ * ]% due upon the
delivery and acceptance of the Certificate of Analysis by the Customer’s QA
Department (less
deposit if required). There is an intention by Advaxis and Cobra to agree on %
royalties of final commercial products utilizing the current Listeria
monocytogenes platform and variations thereof for various indications in
exchange for a reduction in price of the Phase II cGMP manufacturing campaign of
proposal 0422.






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This
phase of the program is governed by the Terms and Conditions set out in the
attached document “ O422 Phase II Terms and Conditions”.







Contract
O422




Determination
of Manufacturing Parameters, Process Development and cGMP Manufacture
of L.
monocytogenes

  




For
Advaxis, Inc.                For Cobra
Biomanufacturing Plc




Accepted
by: J. Todd Derbin          
David R.
Thatcher




Signature:
/s/ J. Todd Derbin   /s/ David
R. Thatcher




Date:
7/7/03  
8th July
2003









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CTL.0422.S.1.1

Provisional
Draft Specification







                      







 







	Test	 Method  Specification
	 	 
	[ * ]	 
	 	 
	
      Identity
	 
	 	 
	
      API
      Listeria
	
      Profile
      number conforms, typically >0.95

	 	 
	
      Growth
      on selective media
	
      Good
      growth

	 	 
	
      Gram
      strain
	
      Gram
      positive

	 	 
	
      Colony
      morphology
	
      Complies
      with that for L. monocytogenes








Quantity




[ *
]




Purity




[ *
]

 

 



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