Document:

Exhibit 4.2(f)

 

“CONFIDENTIAL
TREATMENT REQUESTED. CONFIDENTIAL PORTIONS OF THIS DOCUMENT HAVE BEEN OMITTED
AND HAVE BEEN SEPARATELY FILED WITH THE COMMISSION.  CONFIDENTIAL TREATMENT HAS BEEN REQUESTED
WITH RESPECT TO THE OMITTED PORTIONS.”

 

RESEARCH
SERVICES (SUBCONTRACTING) AGREEMENT

 

AGREEMENT dated 26th June 2000

 

BETWEEN

 

AUTOGEN
RESEARCH PTY LIMITED ACN 074 636 847 of 210 Kings Way, South Melbourne, Victoria 3205 (“Autogen
Research”)

 

AND

 

DEAKIN
UNIVERSITY a body
corporate and politic established pursuant to the Deakin
University Act 1974 of Geelong, Victoria 3217 (“Deakin
University”)

 

RECITALS

 

A.                                   Autogen Research, IDI and Deakin University are
parties to the Research Agreement setting out the terms and conditions for
research (Stage 1 research) to be carried out with the participation of the
parties.

 

B.                                     Pursuant to the Research Agreement, Autogen
Research has granted to Lipha S.A.:

 

•                  a
licence to carry out further (Stage 2) research in respect of an identified
novel gene product arising out of Stage 1 research; and

 

•                  a
licence to use and commercialise the intellectual property emerging from Stage
1 and Stage 2 research.

 

C.                                     Lipha has requested that Autogen Research carry
out specific studies as part of the Stage 2 research into an identified novel
gene product.

 

D.                                    Autogen Research has requested that Deakin
University conduct, and Deakin University agrees to conduct, certain of the
studies referred to in Recital C as a subcontractor and upon the terms and
conditions set out in this Agreement.

 

AGREEMENT

 

1.                                      DEFINITIONS AND INTERPRETATION

 

1.1                               Definitions

 

Unless
an express term of this Agreement or the context otherwise requires, the
following words and expressions have the meanings indicated in this Agreement
(including the Recitals):

 

“Business Day”
means a day on which trading banks are open for general banking business in
Melbourne;

 

“Commencement Date” means the date set out at Item 3 of the Schedule;

 

“Confidential Information” means information that:

 

(a)                                  is by its nature confidential;

 

 

(b)                                 is designated by Autogen Research as
confidential; or

 

(c)                                  Deakin University knows or ought to know is
confidential,

and
includes:

 

(d)                                 information comprised in or relating to the Pre
Stage 2 Results and the Stage 2 Results;

 

(e)                                  information which relates to or affects or may
affect the financial position or reputation of Autogen Research or Lipha;

 

(f)                                    information relating to the
internal management of Autogen Research or Lipha or to the personnel, policies
and strategies of either of them;

 

(g)                                 information of Autogen Research or Lipha to
which Deakin University has access (other than information referred to in
paragraphs (d), (e) and (f)) that has any actual or potential commercial value
to Autogen Research or Lipha;

 

(h)                                 information in the possession of Deakin
University relating to clients or suppliers of Autogen Research or Lipha, and
like information; and

 

(i)                                     information of Autogen Research or Lipha
disclosed to Deakin University before or after the Commencement Date.

 

“Contract
Services” means those
parts of the Stage 2 Research which are to be performed by Deakin University
and which are detailed at Item 7 of the Schedule;

 

“GST” means any goods and services tax which is
levied on the value of goods and services supplied;

 

“IDI” means International Diabetes Institute ACN 007
342 412;

 

“Lipha” means Lipha S.A., a company incorporated under
the laws of France, of 37 rue Saint Romain, 69379 Lyon, CEDEX 08, France;

 

“Novel
Gene Product” means the
novel gene product named or described at Item 2 of the Schedule;

 

“Party” means Autogen Research or Deakin University
and “Parties” means both of them;

 

“Pre-Stage
2 Results” means the
patents and patent applications, and the technical, commercial and other
information, owned by or licensed to Autogen Research related to the Research
Field (other than the Stage 2 Results);

 

“Research
Agreement” means the
Research Agreement dated 28 February 1997 between Autogen Research (then
called Autogen Pty Ltd), Deakin University and IDI as that agreement has since
been renewed and amended;

 

“Research
Field” means the
research field named at Item 1 of the Schedule;

 

“Services
Fee” means the fee
payable to Deakin University for performance of the Contract Services, being
the amount set out at Item 5 of the Schedule;

 

 

“Stage 2
Results” means the
patents and patent applications, and the technical, commercial and other information
relating to or arising out of Stage 2 Research;

 

“Stage 2
Research” means certain
research in respect of the Novel Gene Product which Lipha is undertaking or
will undertake, and in respect of which Lipha has appointed Autogen Research to
carry out specific studies, and of which the Contract Services are a component;

 

“Term” means the 12 month period beginning on the
Commencement Date.

 

1.2                               Interpretation

 

In
this Agreement, headings are for ease of reference only and do not affect the
meaning of this Agreement, and unless the contrary intention appears:

 

(a)                        the singular includes the plural and vice versa and words importing a gender include other genders;

 

(b)                       other grammatical forms of defined words or
expressions have corresponding meanings;

 

(c)                        a reference to a clause, paragraph, schedule or
annexure is a reference to a clause or paragraph of, or schedule or annexure
to, this Agreement and a reference to this Agreement includes any schedules and
annexures;

 

(d)                       a reference to a Party includes its successors
and permitted assigns;

 

(e)                        words and expressions importing natural persons
include partnerships, bodies corporate, associations, governments and governmental
and local authorities and agencies; and

 

(f)                          a reference to dollars and “$” means the lawful
currency of Australia.

 

2.                                      TERM OF AGREEMENT

 

2.1                                 Subject to any early termination under clause
9, this Agreement will be effective during the Term.

 

2.2                                 Autogen Research and Deakin University may at
any time extend the Tein-1 for any further period or periods by
agreeing upon the Contract Services to be provided by Deakin University during
such extended term and the Services Fee payable by Autogen Research during the
extended term.

 

3.                                      PROVISION OF CONTRACT SERVICES BY
DEAKIN UNIVERSITY

 

3.1                                 During the Term Deakin University will provide
the Contract Services to or at the direction of Autogen Research.

 

3.2                                 Deakin University will provide the services of
a suitably qualified and experienced person or persons to supervise and direct
the scientific staff engaged in the conduct of the Contract Services.

 

3.3                                 The Contract Services will be carried out at
the School of Nutrition and Public Health, Deakin University, Pigdons Road,
Geelong, Victoria, or at such other place as Deakin University may nominate,
and Autogen Research may agree, from time to time.

 

 

3.4                                 Deakin University shall employ or engage on
contract such scientific staff as are necessary to provide the Contract
Services.

 

3.5                                 Deakin University may subcontract the Contract
Services only with the prior written consent of Autogen Research, such consent
to be given or withheld at Autogen Research’s absolute discretion and subject
to such conditions as Autogen Research thinks fit.

 

3.6                                 Deakin University shall at all times use its
best endeavours to ensure that completion of each part of the Contract Services
is achieved generally in a timely and efficient manner and particularly in
accordance with any timetable or “milestone” dates specified at Item 7 of the
Schedule.

 

3.7                                 Deakin University shall at all times indemnify,
hold harmless and defend Autogen Research and its officers, employees and
agents (in this clause 3.7 referred to as the “Indemnified Parties”) from and
against any loss (including reasonable legal costs and expenses) or liability
reasonably incurred or suffered by any of the Indemnified Parties arising from
any suit, action or proceeding by any person against any of the Indemnified
Parties where such loss or liability was caused by any wilful, unlawful or
negligent act or omission of Deakin University, its employees, agents or
sub-contractors in connection with this Agreement.

 

3.8                                 Deakin University covenants and undertakes that
the Contract Services, including any tests and trials carried out as part of
the Contract Services, will at all times be conducted to the highest possible
professional standards and in accordance with all applicable rules, regulations
and conditions and in particular will procure that no actions, suits or
proceedings will be made against Autogen Research in respect of or in
connection with the conduct of the Contract Services.

 

3.9                                 Deakin University shall be responsible for
obtaining necessary regulatory approval (if any) by any and all government
agencies for conducting the Contract Services in Australia. Deakin University
shall deliver copies of all such approvals (if any) to Autogen Research within
7 days of receipt of such approvals by Deakin University.

 

4.                                      REPORTING BY DEAKIN UNIVERSITY

 

4.1                                 Deakin University will prepare and submit a
report to Autogen Research every 3 months during the Term which sets out in
reasonable and informative detail:

 

(a)                        the progress of the Contract Services during
the period to which the report relates, and whether all milestones which ought
to have been reached during that period have been reached and best professional
standards maintained;

 

(b)                       any material advances or developments;

 

(c)                        any material delays or unforeseen problems in the
conduct of the Contract Services;

 

(d)                       any recommendations on changes to the Contract
Services; and

 

(e)                        any other relevant information relating to or
affecting the Contract Services.

 

4.2                                 Reports required under clause 4.1 will be
provided by Deakin University to Autogen Research on the same dates and in
relation to the same periods as the reports to be provided

 

 

under clause 5.3 of the
Research Agreement, unless Autogen Research nominates otherwise in writing to
Deakin University.

 

4.3                                 In addition to the reports under clause 4.1,
Deakin University will immediately advise Autogen Research in writing of any
material advancements or developments which arise out of the performance of the
Contract Services.

 

5.                                      PAYMENT

 

5.1                                 Services Fee

 

(a)                        Autogen Research will pay to Deakin University
the Services Fee in such instalments and at such times as are specified at Item
6 of the Schedule..

 

(b)                       Deakin University will invoice Autogen Research
for each instalment of the Services Fee which is payable under paragraph (a) not
less than 30 days before the due date for such payment.

 

5.2                                 Goods and Services Tax

 

(a)                        If at any time during the Term there is any new
GST introduced in Australia, or if there is any change to any GST then in force
or the method of its imposition, then representatives of the Deakin University
and Autogen Research will meet to renegotiate the prices applying to provision
of the Contract Services.

 

(b)                       The tem-is of this Agreement
(including, but not limited to, the Services Fee) will continue to apply until
the renegotiation is completed and the Parties agree in writing to any
amendment of those terms.

 

(c)                        In any renegotiation of the Services Fee which
occurs pursuant to paragraph (a), the Parties will apply the following
objectives:

 

(i)                                     Deakin University should recover from Autogen
Research amounts equal to any liability to remit GST incurred by Deakin
University because of the provision of the Contract Services to Autogen
Research; and

 

(ii)                                  Autogen Research should recover from Deakin
University amounts equal to any reduction in the cost to Deakin University of
providing the Contract Services to Autogen Research where such reduction is a
result of the GST or of the associated abolition or change in rate or method of
imposition of any other tax, excise, duty or impost.

 

(d)                       If within 30 Business Days of the first meeting
of representatives under paragraph (a) Autogen Research and Deakin
University have not been able to agree on the Services Fee
which will apply to the provision of the Contract Services, then either Party
may, by giving notice of not less than 30 Business Days to the other, terminate
this Agreement.

 

6.                                      [NOT USED]

 

7.                                      CONFIDENTIALITY

 

7.1                                 All information relating to the Contract
Services which is supplied by or on behalf of Autogen Research or which relates
to or arises from the Contract Services shall be treated by Deakin University
as confidential and shall be used solely during the Term and in

 

 

accordance with this Agreement
to enable Deakin University to carry out its obligations under this Agreement.

 

7.2                                 Prior to disclosing any of the Confidential
Information to any of its employees, related entities, consultants or
sub-contractors, Deakin University will ensure that the person in question
executes a deed, in form and substance acceptable to Autogen Research,
undertaking to keep secret and confidential the Confidential Information and
acknowledging the interest of Autogen Research therein.

 

7.3                                 Deakin University hereby undertakes that,
except for such disclosure as is prudent and reasonably necessary for the
purposes of this Agreement, no part of the Confidential Information given to it
pursuant to this Agreement will be disclosed to anyone who is not an employee
or subcontractor (as approved by Autogen Research under clause 3.5) of Deakin
University except with Autogen Research’s prior written approval, which if
given shall be on the basis that the recipient of any part of the Confidential
Information shall first be bound to Deakin University and Autogen Research by
contract to maintain the same in confidence. Without prejudice to the
foregoing, Deakin University shall use its best endeavours to prevent the
Confidential Information from passing into the public domain.

 

7.4                                 Nothing stated herein shall be construed as
restricting or creating any liability for the disclosure or communication of
Confidential Information which:

 

(a)                        is now or becomes publicly known through no
wrongful act of Deakin University;

 

(b)                       is received from a third party without
restriction and without breach of this Agreement;

 

(c)                        is now or comes to be contained in any
published patent or published or otherwise generally known to the trade through
no wrongful act of Deakin University; or

 

(d)                       is disclosed pursuant to governmental,
legislative or judicial requirement, including disclosure by Autogen Limited
ACN 000 248 304 (the parent company of Autogen Research) pursuant to its
obligations under the Corporations Law or the Stock Exchange listing rules.

 

7.5                                 The obligations set out in this clause 7 shall
remain in full force and effect, and shall continue to bind each Party,
notwithstanding that this Agreement may have been terminated for any reason.

 

8.                                      OWNERSHIP AND USE OF INTELLECTUAL
AND INDUSTRIAL PROPERTY

 

8.1                                 Deakin University acknowledges that:

 

(a)                        in relation to the Pre-Stage 2 Results:

 

(i)                                     Autogen Research is the owner or the exclusive
licensee of the Pre-Stage 2 Results and has granted to Lipha (among other
rights) a licence of the Pre-Stage 2 Results for research purposes, such
licence being exclusive in the Research Field; and

 

(ii)                                  Lipha has granted to Autogen Research a
non-exclusive royalty free right to use the Pre-Stage 2 Results for the purpose
of performing the Stage 2 Research; and

 

 

(b)                       in relation to the Stage 2 Results - the Stage
2 Results will belong solely to Lipha, and Lipha has granted to Autogen
Research (among other rights) a licence to use the Stage 2 Results for the
purpose of performing the Stage 2 Research.

 

8.2                                 Autogen Research grants to Deakin University a
non-exclusive royalty free light to use:

 

(a)                                  the Pre-Stage 2 Results; and

 

(b)                                 the Stage 2 Results,

 

for the sole purpose of
providing the Contract Services and otherwise carrying out Deakin University’s
obligations under this Agreement.

 

8.3                                 Deakin University may grant a sublicence of the
rights referred to in clause 8.2 to any subcontractor approved by Autogen
Research in accordance with clause 3.5.

 

9.                                      INDEMNITIES AND INSURANCE

 

9.1                                 Autogen Research will indemnify, defend and
save harmless Deakin University, its council, officers and employees from and
against all liabilities, damages, losses, penalties, demands, suits, costs,
expenses (including reasonable lawyers’ fees) and proceedings of any nature or
claims by any person arising out of or in connection with this Agreement, but
only to the extent to which the same is caused by or contributed to by the
negligent or intentional act or omission of Autogen Research or its directors,
officers, employees or agents.

 

9.2                                 Deakin University will indemnify, defend and
save harmless Autogen Research, its directors, officers and employees from and
against all liabilities, damages, losses, penalties, demands, suits, costs,
expenses (including reasonable lawyers’ fees) and proceedings of any nature or
claims by any person arising out of or in connection with this Agreement, but
only to the extent to which the same is caused by or contributed to by the
negligent or intentional act or omission of Deakin University or its directors,
officers, employees or agents.

 

9.3                                 At all times during the Term each Party must,
at their own expense, establish and keep current, and Deakin University must
ensure that any person to whom the Contract Services are subcontracted pursuant
to clause 3.5 establishes and keeps current, the insurance policies of standard
form, and subject to exclusions and sub-limits appropriate for this Agreement,
and otherwise as specified in clauses 9.4 and 9.5.

 

9.4                                 The policies of insurance required under clause
9.3 are as follows:

 

(a)                        public liability insurance with a limit of
liability of not less than $20 million for any one occurrence;

 

(b)                       product liability insurance with a limit of
liability of not less than $20 million for any one occurrence and in the annual
aggregate;

 

(c)                        workers’ compensation insurance in accordance
with the laws in force in Victoria; and

 

(d)                       professional indemnity insurance with a limit
of liability of not less than $15 million for any one claim made. Professional
Indemnity cover is to be maintained throughout the term and until 7 years after
the end of the term.

 

9.5                                 Each policy of insurance referred to in this
clause 9 must:

 

 

(a)                        be made available for inspection by each Party
at any time during the Term upon the reasonable request of the other Party,
together with certificates evidencing those policies, the payment of premiums
and other reasonable documentation which confirms that the policies are valid,
current and meet the requirements of this Agreement;

 

(b)                       name Lipha, Autogen Research and their
employees, contractors and agents as coinsured persons so that the policy will
respond as if there is a separate contract of insurance between each co-insured
person and the insurer;

 

(c)                        include a clause enabling one insured person to
claim against the insurer where another insured person would have been entitled
to claim against the insurer but is precluded from doing so for any reason
including, but not limited to, a breach of the policy by that other insured
person; and

 

(d)                       include a cross-liability clause enabling one
insured person to claim against the insurer even if the person making the claim
against that insured person is also insured under the policy.

 

9.6                                 If a Party (“Defaulting Party”)
fails to effect or to keep in force any of the insurances which it is required
by this Agreement to effect and maintain, the other Party (“Other
Party”) may, but shall not be obliged to:

 

(a)                        effect and keep in force any such insurance and
pay such premiums and other moneys as may be necessary for that purpose and may
recover as a debt due from the Defaulting Party on demand the amount so paid
plus an administration charge of 10%; or

 

(b)                       refuse to perform its obligations under this
Agreement until the insurance policies and a receipt for the payment of
premiums are made available by the Defaulting Party for inspection.

 

Notwithstanding
that the Other Party may effect such insurances, the Defaulting Party will be
deemed to have indemnified the Other Party against all claims, demands,
proceedings, costs, charges and expenses which may arise as a result of
the Defaulting Party’s failure to so insure.

 

10.                               TERMINATION

 

10.1                           The occurrence of any of the following events
with respect to Autogen Research or Deakin University constitutes a material
default (“Material Default”) in respect of that
Party:

 

(a)                        failure by the Party to make when due any
payment under this Agreement if that failure is not cured within 20 Business
Days after notice of that failure is given to the Party by the other Party;

 

(b)                       failure by the Party to comply with any
obligation (other than an obligation to pay) under this Agreement if that
failure is not remedied or its effect mitigated to the reasonable satisfaction
of the other Party within 20 Business Days after notice of that failure is
given to the Party by the other Party, unless the Party has commenced to remedy
the failure during that period of 20 Business Days and continues to use its
best efforts in that regard to the reasonable satisfaction of the other Party;

 

(c)                        the Party:

 

 

(i)                                     becomes insolvent or is unable to pay its debts
when they fall due;

 

(ii)                                  is dissolved, wound up, placed under official
management, liquidated or other administration or arrangement is made or
passed;

 

(iii)                               seeks or becomes subject to the appointment of a receiver, receiver and
manager, liquidator, provisional liquidator, trustee, administrator, custodian
or similar person (“Appointee”),
unless the Appointee is removed within 5 Business Days or is appointed to
assets of the Party which do not comprise a substantial part of its assets as a
whole; or

 

(iv)                              makes a general assignment, arrangement or
composition with or for the benefit of its creditors;

 

(d)                       the Party or any validly appointed Appointee disaffirms
or repudiates this Agreement.

 

10.2                           If a Material Default with respect to a Party (“Defaulting Party”) has occurred and is continuing, then
the other Party may give a notice to the Defaulting Party:

 

(a)                        specifying the relevant Material Default; and

 

(b)                       designating a. day not less than 10 Business
Days from the date the notice is given as the termination date.

 

10.3                           If a notice is given pursuant to clause 10.2
then this Agreement will terminate on the date designated in accordance with
clause 10.2(b) if the Material Default is not remedied or the Defaulting
Party has not commenced to remedy the Material Default and continues to use its
best efforts to do so to the reasonable satisfaction of the other Party.

 

11.                               INCORPORATION OF FURTHER TERMS BY
REFERENCE

 

Clauses
8.4, 9 (except for clause 9.5), 11 (except for clause 11.4), 12, 13 and 14 of
the Research Agreement are incorporated into and read together with this
Agreement as if they were duplicated herein, save that:

 

(a)                        each reference therein to “Autogen” is deleted
and replaced with a reference to “Autogen Research”;

 

(b)                       each reference therein to “the Project” is
deleted and replaced with a reference to “the Contract Services”; and

 

(c)                        in clause 14.1(b)(ii), the number for facsimile
transmission of a notice to Autogen Research is deleted and replaced with “9234
1255”.

 

12.                               ASSIGNMENT

 

This
Agreement is personal to the Parties and neither Party may transfer, assign,
subcontract or encumber, in whole or in part, its interest in this Agreement
without the prior mitten consent of the other, which consent may be given or
withheld in the absolute discretion of the other Party, and upon such
conditions as the other Party may think fit.

 

13.                               GENERAL

 

13.1                           Entire Agreement

 

 

Subject
to any applicable provisions of the Research Agreement, this Agreement
constitutes the entire agreement of the Parties in relation to the Contract
Services and all prior representations, communications and agreements in
relation to the subject matter of this Agreement are merged in or suspended by
this Agreement.

 

13.2                           Amendment

 

This
Agreement may only be amended or supplemented in writing signed by the Parties.

 

13.3                           Survival of indemnities and other provisions

 

Each
indemnity in this Agreement and each provision which is expressed to survive
termination of this Agreement is a continuing obligation which survives
termination of this Agreement.

 

13.4                           Counterparts

 

This
Agreement, and any amendment in respect of it, may be executed and delivered in
counterparts, including by facsimile transmission.

 

13.5                           Nature of Agreement

 

This
Agreement does not create any relationship of agency, partnership, employment,
joint venture or legal representation between the Parties. Autogen Research and
Deakin University acknowledge that they are independent contracting parties and
do not have any authority or power for or on behalf of each other to enter into
any contract, incur debt, accept money, assume obligations or make any
warranties or representations.

 

	
  EXECUTED as an Agreement

  	
   

  	
   

  
	
  THE
  COMMON SEAL of

  	
  )

  	
   

  
	
  AUTOGEN
  RESEARCH PTY

  	
  )

  	
   

  
	
  LIMITED
  ACN 074 636 847 was

  	
  )

  	
   

  
	
  hereunto
  affixed in accordance with

  	
  )

  	
   

  
	
  its
  Constitution in the presence of:

  	
  )

  	
   

  
	
   

  	
   

  	
   

  
	
   

  	
   

  	
   

  
	
   

  	
   

  	
   

  	
   

  
	
  Secretary

  	
   

  	
  Director

  
	
   

  	
   

  	
   

  
	
  THE
  COMMON SEAL of

  	
  )

  	
   

  
	
  DEAKIN
  UNIVERSITY  was

  	
  )

  	
   

  
	
  hereunto
  affixed on the 26th day of June

  	
  )

  	
   

  
	
  by
  direction of theVice-Chancellor

  	
  )

  	
   

  
	
  in
  the presence of.

  	
  )

  	
   

  
	
   

  	
   

  	
   

  
	
   

  	
   

  	
   

  
	
   

  	
   

  	
   

  	
   

  
	
  Vice-Chancellor

  	
   

  	
  Vice-President
  (Administration)

  

 

 

SCHEDULE

 

	
  ITEM 1 -
  The Research Field:

  	
   

  	
  Obesity

  
	
   

  	
   

  	
   

  
	
  ITEM 2 -
  The Novel Gene Product:

  	
   

  	
  Beacon

  
	
   

  	
   

  	
   

  
	
  ITEM 3 -
  The Commencement Date:

  	
   

  	
  TERM
  1

  	
  1
  April 1999 

  
	
   

  	
   

  	
   

  	
   

  
	
   

  	
   

  	
  TERM
  2:

  	
  1
  April 2000

  
	
   

  	
   

  	
   

  	
   

  
	
  ITEM 4 -
  NOT USED

  	
   

  	
   

  	
   

  
	
   

  	
   

  	
   

  	
   

  
	
  ITEM 5 -
  The Research Services Fee:

  	
   

  	
  TERM
  1

  	
  $[*]

  
	
   

  	
   

  	
   

  	
   

  
	
   

  	
   

  	
  TERM
  2:

  	
  $[*]

  
	
   

  	
   

  	
   

  	
   

  
	
  ITEM 6 -
  Payment of Research Services Fee:

  	
   

  	
  by
  4 equal quarterly instalments in advance, the first instalment being due and
  payable on the Commencement Date in respect of each Term and subsequent
  instalments being due and payable 3 months, 6 months and 9 months after the
  Commencement Date in respect of each Term

  
	
   

  	
   

  	
   

  
	
  ITEM 7 -
  The Contract Services:

  	
   

  	
  Refer
  to the following 23 pages

  

 

 

Stage 2       1999-2002

 

OBESITY
GENES RESEARCH PLAN

 

 

Metabolic
Research Unit

 

 

DEAKIN
UNIVERSITY

 

 

Research
Head:

Professor
Greg R. Collier

 

 

OBESITY
GENES RESEARCH PLAN

 

The
current Stage I obesity research plan is aimed at the discovery of new genes
involved in the development of obesity. From this program it is anticipated
that 3 key genes will proceed to Stage II research over the next 3 years, the
first to begin Stage II will be beacon.

 

Stage
II research will involve research at Deakin University as well as Lipha. Stage
II includes determining an appropriate cell model and animal model for
screening beacon antagonists, high throughput screening, clinical development
and clinical trials.

 

It
is anticipated that initial studies that include localization of site of
cell/tissue action and development of the animal model and the in vitro cell
system for screening will be conducted at Deakin University.

 

These
beacon Stage II studies with Deakin are anticipated to be completed in 18
Illol7ths. However, gene 2 and gene 3 studies will begin in April 2000 and
April 2001, respectively. The budget proposed will be an annual budget for
3 years and will cover the studies for all of the 3 genes proposed. We will
select genes closely related to beacon or expressed in hypothalamus so that
existing technology will be utilised for all 3 genes.

 

It
is anticipated that Lipha will he responsible for all high throughput
screening, clinical development and clinical trials. Deakin will provide Lipha
with results for beacon after 18 months and the next 2 genes at 18 month
intervals for future studies.

 

 

BEACON STAGE 11 RESEARCH PLAN

 

	
  April 1999

  	
   

  	
  April 2000

  	
   

  	
  October 2000

  	
   

  	
  October 2001

  	
   

  	
  July 2002

  

 

 

	
  In vivo studies – Deakin

  	
   

  	
  Lipha Studies

  	
   

  	
   

  	
   

  
	
   

  	
   

  	
   

  	
   

  	
   

  	
   

  
	
  A

  	
   

  	
  Miniaturization
  of cellular test

  	
   

  	
   

  	
   

  
	
  B

  	
   

  	
   

  	
   

  	
   

  	
   

  
	
  C

  	
   

  	
  High throughput screening

  	
   

  	
   

  	
   

  
	
   

  	
   

  	
   

  	
   

  	
   

  	
   

  
	
   

  	
   

  	
  Confirmed hits

  	
   

  	
   

  	
   

  
	
  If
  successful at. this point, we will have an animal model suitable
  for screening antagonist and potentially an

  	
   

  	
   

  	
   

  	
   

  	
   

  
	
  obesity
  diagnostic test~

  	
   

  	
  In vivo studies.

  	
   

  	
   

  	
   

  
	
  In
  vitro studies - Deakin .

  	
   

  	
  In vitro studies

  	
   

  	
   

  	
   

  
	
   

  	
   

  	
   

  	
   

  	
   

  	
   

  
	
  D

  	
   

  	
   

  	
   

  	
   

  	
   

  
	
  E

  	
   

  	
   

  	
   

  	
   

  	
   

  
	
  F

  	
   

  	
  Leads

  	
   

  	
   

  	
  lead
  compound

  
	
  G

  	
   

  	
   

  	
   

  	
   

  	
   

  
	
   

  	
   

  	
   

  	
   

  	
   

  	
   

  
	
   

  	
   

  	
  if
  successful at this stage we will have a cell system available for in vitro
  testing of either beacon promoter, partner or receptor antagonist

  	
   

  	
   

  
										

 

 

COMPLETE STAGE II DEAKIN - 3 YEAR OBESITY PLAN

 

	
  Apri1
  1999

  	
   

  	
  April 2000

  	
   

  	
  April 2001

  	
   

  	
  April 2002

  

 

Beacon
gene

 

In
vivo studies

 

In vitro studies

 

New obesity gene A

 

It
is important to. note that the introduction of new genes into Stage
II will complement the technology already established with beacon. This will
mean that obesity genes isolated in the hypothalamus .(and possibly related to
beacon) will be given priority.. This will ensure the annual budget for Stage
11 will be the same over the 3 years.

 

In vivo studies

 

In vitro-studies

 

	
   

  	
  New obesity gene B

  	
  

  
	
   

  	
  In vivo studies

  
	
   

  	
   

  
	
   

  	
  In vitro-studies

  

 

 

BEACON
RESEARCH PLAN 1999 - 2000

 

Aims:

 

Animal
Studies

 

A.                To
find a suitable animal model to screen beacon antagonists.

 

B.                  To
find tissue sites of beacon action and to find intracellular-
localization/action.

 

C.                  To
establish whether beacon is a circulating protein.

 

Molecular and Cellular Studies

 

D.                 To find the beacon receptor from existing cDNA
libraries.

 

E.                   To identify beacon receptors in tissue culture
cell lines.

 

F.                   To identify a beacon ‘partner’ rather than a
receptor.

 

G.                  To
find the beacon promoter sequence and construct a cell system for regulating
promoter activity.

 

 

STAGE II - ANIMAL STUDIES —
DEAKIN

 

A.
Beacon action in other animal models.

 

Aim: To find a suitable animal model to screen beacon. ISR model not
suitable for large screening studies due to difficulty in animal supply
handling. It will also give more information on beacon in other animal models.          .         .

 

	
  (a)

  	
   

  	
  Check
  beacon hypothalamus-gene expression in, many animal models and also sequence
  beacon gene (ob/ob, db/db, NZO, ZDF, SHR, Sprague-Dawley). Priority will be
  given to animal models commonly available.

  
	
   

  	
   

  	
   

  
	
  (b)

  	
   

  	
  Sprague-Dawley
  rats

  	
  

  	
  ICV/IP/IV

  	
  

  	
  NO
  EFFECT

  	
  

  	
  Go to
  other animal models, see

  	
   

  
	
   

  	
   

  	
   

  	
   

  	
  

  	
   

  	
  

  	
  

  	
   

  
	
   

  	
   

  	
   

  	
   

  	
  EFFECT

  	
   

  	
   

  	
   

  	
  NO EFFECT

  	
   

  
	
   

  	
   

  	
   

  	
   

  	
  

  	
   

  	
   

  	
   

  	
   

  	
   

  
											

 

IDENTIFIED SUITABLE ANIMAL MODEL
FOR SCREENING POTENTIAL BEACON ANTAGONISTS (it is anticipated that the
Sprague-Dawley and screening animal model studies will take 3 months and 12
months, respectively)

 

 

B.                                    Beacon
localization/sites of action.

 

Aim: to find  tissue sites of beacon action and to find intracellular
localization/action

 

	
  (a) Israeli Sand rats (A  and C animals)

  	
   

  	
  I125-beacon

  	
  

  	
  localization (in vivo and in vitro)/tissue uptake

  
	
   

  	
   

  	
   

  	
   

  	
   

  
	
  * IF NO HYPOTHALAMUS UPTAKE*

  	
  

  	
  DECISION

  	
  

  	
  ACTIVITY OF I125 BEACON?

  
								

 

	
  C. Western blots for quantitation of protein levels

  	
   

  	
   

  
	
  

  	
  

  	
   

  	
   

  
	
  Human protein medleys (Clontech)

  	
   

  	
  ISR tissues (compare to mRNA)

  
				

 

* WILL PROVIDE
INFORMATION 0N TISSUES IN WHICH TO STUDY BEACON ACTION *

 

	
  (c)  Israeli Sand rats (A and C animals)

  	
   

  
	
  

  	
  

  	
   

  
	
  Immunohistochemistry

  	
  In situ hybridization

  
				

 

*USED AS A QUICK SCAN TO IDENTIFY TISSUES OF
INTEREST? *

 

 

C.                                    Is
beacon a circulating protein?

 

Aim: to establish whether beacon is
circulating or not

 

(a)               In vitro cell
production — test both media and cell lysates by Western

 

(b)              Sera from humans and
animals — test by Western and establish ELISA

 

	
  Yes, beacon is circulating

  	
   

  	
  No, beacon is not circulating

  
	
   

  	
   

  	
   

  
	
  

  	
   

  	
  

  
	
   

  	
   

  	
   

  
	
  Continue with receptor search!

  	
   

  	
  Should we continue with beacon receptor work?

  

 

 

STAGE II — MOLECULAR AND CELLULAR STUDIES — DEAKIN

 

D.                                    Identification
of beacon receptor

 

Aim: These studies are based on the
assumption that beacon is a circulating protein and binds to its own receptor.
The aim is to find and clone the beacon receptor from cDNA libraries by
expression cloning.

 

(a)                                  Receptor
panning and sib selection

 

	
  NO EXPRESSION

  	
  

  	
  human brain cDNA library

  	
   

  	
   

  
	
  

  	
  

  	
  

  	
   

  
	
   

  	
  ISR hypothalamus cDNA library

  	
  

  	
  RECEPTOR EXPRESSION

  	
   

  
	
   

  	
  

  	
   

  	
  

  
	
  Wrong tissue?

  	
   

  	
   

  	
   

  	
   

  	
   

  
	
   

  	
  

  	
   

  	
   

  	
   

  	
   

  	
   

  	
   

  
	
   

  	
   

  	
   

  	
  Check

  receptor

  expression

  in cell

  lines

  	
   

  	
  Transfect

  receptor

  into CHO

  cells

  	
   

  
	
   

  	
   

  	
   

  	
   

  	
  

  	
   

  	
  Screen

  	
   

  
	
   

  	
   

  	
   

  	
   

  	
   

  	
  

  	
   

  
	
   

  	
   

  	
   

  	
   

  	
   

  	
   

  	
   

  
	
   

  	
   

  	
  CELL MODEL AVAILABLE FOR SCREENING

  BEACON RECEPTOR ANTAGONISTS

  	
   

  
											

 

 

E.                                      Cell
culture

 

Aim: to identify and clone beacon receptor in
cell lines and to also study signal transduction mechanisms

 

 (a)                               Cell
lines include 3T3, GT1-7, HepG2, primary cultures from ISR

 

	
   

  	
   

  	
   

  	
  Effect of modulation of beacon expression
  in cell lines (page

  11)

  	
   

  
	
   

  	
   

  	
   

  	
   

  	
   

  
	
  Treat cells with beacon ± NPY, beacon ± leptin

  	
  

  	
  NO EFFECT

  	
  

  	
   

  	
   

  
	
   

  	
  

  	
   

  	
   

  	
   

  
	
   

  	
  OBSERVED EFFECT

  	
   

  	
   

  
	
   

  	
   

  	
  

  	
   

  	
   

  
	
   

  	
   

  	
  Macro/microarrays

  	
   

  	
   

  
	
   

  	
   

  	
  

  	
   

  	
   

  
	
   

  	
  Receptor cloning from cDNA

  	
   

  	
   

  	
  Select 3-5 genes (if GT-1-7 cells, test for GnRH)

  
	
   

  	
   expression library of cell line

  	
   

  	
  

  	
   

  	
  

  	
   

  
	
   

  	
   

  	
   

  	
   

  	
  Real-time PCR

  

  	
   

  
	
   

  	
   

  	
   

  
	
   

  	
  Signal transduction studies

  	
   

  	
   

  	
   

  	
   

  	
   

  	
   

  	
   

  	
   

  	
   

  
	
   

  	
   (Ca2+, cAMP, kinases, phosphatase)

  	
   

  	
  

  	
  YES

  	
  

  	
  Time
  course experiments

  	
  

  	
  NO

  	
  

  	
  2 hybrid system

  
	
  

  	
   

  	
   

  	
   

  	
   

  	
   

  	
  (page 12) and

  
	
   

  	
   

  	
   

  	
   

  	
   

  	
   

  	
  beacon promoter (page 13)

  
	
   

  	
  STOP STUDIES LISTED UNDER D., F.
  AND G

  	
   

  	
   

  	
   

  	
   

  	
   

  
														

 

 

	
  IF NO EFFECT IN CELL LINES WITH BEACON TREATMENT

  
	
  

  	
   

  	
   

  	
   

  	
  

  	
   

  
	
  Overexpression of beacon in cells by transfection (BacMan,
  adenovirus)

  	
   

  	
   

  	
  Neutralisation of antisense treatment

  	
   

  
	
  

  	
   

  	
   

  	
  

  
	
   

  	
  Find leptin sensitive cell line

  	
   

  
	
   

  	
  

  	
   

  
	
   

  	
  Immunoneutralisation of beacon

  	
   

  
	
   

  	
  

  	
   

  
	
   

  	
  Back to macro/microarrays (section E,

  page 10)

  	
   

  
	
   

  	
   

  	
  

  	
   

  	
   

  
	
   

  	
   

  	
  Select 3-5 genes

  	
   

  	
   

  
	
   

  	
   

  	
  

  	
   

  	
   

  
	
   

  	
   

  	
  Time course experiments

  	
  

  	
  NO EFFECT

  	
  

  	
  Beacon

  
	
   

  	
   

  	
  

  	
   

  	
   

  	
   

  	
  promoter

  (page 13)

  
	
   

  	
   

  	
  EFFECT

  	
   

  	
   

  	
   

  	
   

  
	
   

  	
   

  	
  

  	
   

  	
   

  	
   

  	
   

  
	
   

  	
  SECONDARY CELL SYSTEM FOR SCREENING

  ANTAGONISTS OF BEACON PARTNERS AND

  MODULATORS OF THE BEACON PROMOTER

  	
   

  
											

 

 

F.                                      2 hybrid system

 

Aim: if beacon is not secreted to look for beacon “partner rather than
receptor

 

	
   

  	
   

  	
  Yeast two-hybrid system

  	
   

  	
   

  
	
   

  	
   

  	
  

  	
   

  	
   

  
	
  If partner is unknown

  	
  

  	
  Identify beacon partner

  	
   

  	
   

  
	
  

  	
   

  	
  

  	
  

  	
   

  	
   

  
	
  Beacon promoter?

  (page 13)

  	
   

  	
  Transfect into CHO cells

  	
   

  	
   

  	
  If the partner is a known protein implicated in signal transduction
  pathways

  
	
   

  	
   

  	
  Screen

  	
   

  	
   

  	
  

  
	
   

  	
   

  	
   

  	
   

  
	
   

  	
   

  	
   

  	
   

  	
  In vitro tests

  
	
   

  	
  CELL SYSTEM FOR SCREENING

  ANTAGONISTS OF BEACON

  PARTNERS

  	
   

  	
   

  	
   

  
									

 

 

	
  G.

  	
   

  	
  Beacon
  promoter

  
	
   

  	
   

  	
   

  
	
   

  	
   

  	
  Aim:
  to identify and clone the beacon promoter and construct a cell line model for
  screening/regulating promoter

  
	
   

  	
   

  	
   

  	
   

  	
   

  	
   

  	
   

  
	
  (a)

  	
   

  	
  Promoter
  cloning and sequencing

  	
   

  	
  

  	
   

  	
  sequence
  analysis

  
	
   

  	
   

  	
   

  	
   

  	
   

  	
   

  	
   

  
	
   

  	
   

  	
   

  	
   

  	
  

  	
   

  	
  promoter
  activity

  
	
   

  	
   

  	
   

  	
   

  	
   

  	
   

  	
   

  
	
  (b)

  	
   

  	
  Reporter
  gene assay

  	
   

  	
   

  	
   

  	
   

  
	
   

  	
   

  	
   

  	
   

  	
   

  	
   

  	
   

  
	
  (c)

  	
   

  	
  Transfect
  different cell lines

  	
  

  	
   

  	
  CELL
  MODEL FOR SCREENING THE REGULATION OF BEACON PROMOTER

  

 

 

From page 10

 

EFFECT

 

Receptor cloning

Signal Transduction studies

 

CELL SYSTEM AVAILABLE FOR SCREENING

ANTAGONISTS OF BEACON RECEPTOR

 

 

In Vivo Studies

 

1.                          Tissue
distribution of 1251-beacon.

 

In these studies, lean and obese Israeli Sand rats will be used. It is
anticipated that 125I- labelled beacon will be injected
intracerebroventricularly, intravenously and intra peritoneally.

 

These studies will provide information on uptake and clearance of
beacon protein. It is anticipated that these studies will be repeated when the
full-length recombinant beacon protein becomes available.

 

Beacon tissue distribution and plasma
clearance (method modified from Hill et al. (1997)

 

125I-beacon (Sx106c.p.ln)
with a specific radioactivity of 160μCi/μg, (prepared by the iodogen method)
will be injected intracerebroventricularly, intravenously and intra
peritoneally into lean and obese Israeli Sand Rats. The jugular vein of each
rat will be canulated and blood samples (200μ1) will be collected at regular
intervals between t=0min to t=180min. The heparinised blood samples will then
be centrifuged and plasma stored at -20°C.

 

At the end of the experiment rats will be killed by cervical
dislocation and the following tissues collected, weighed and frozen in liquid
nitrogen: brain, hypothalamus, spleen, liver, pancreas, kidneys, heart, lungs,
reproductive organs, mesentery, small intestines, fat (perirenal, epididymal,
sub scapular), and muscle (gastrocnemius, soleus, plantaris). Tissues will then
be stored at -20°C. Urine will be obtained from the bladder and
stored at -20°C.

 

Total and trichloroaceteic acid (TCA)-precipitable material, which is
representative of undegraded beacon, will be measured in all samples. Duplicate
portions (25 μ1) of the plasma (from each time interval) and urine samples will
be mixed with 475 μ1 ice-cold 10% trichloroacetic acid (TCA), incubated for 2 h
at 0°C and centrifuged. The radioactivity in both the TCA-soluble
and TCA-insoluble fractions will be measured. The frozen tissue samples will be
homogenised in 10% (w/v) TCA, kept on ice for 1h to precipitate proteins,
centrifuged (3000g, 20min, 4°C) and total precipitable radioactivity
counted.

 

Beacon: localisation in the brain

 

In vivo autoradiography:

 

Two, 10 or 30min after ICV injection of 125I-beacon (5x106c.p.m)
lean and obese rats will be decapitated and the brains removed. The intact
brains will be immediately frozen in liquid nitrogen and then stored for 24 h
at -70°C. Frozen brains will then be warmed to -15°C, coronally
s1iced at 20μm and mounted on gelatin coated slides.

 

The mounted sections will be desiccated for 24 11 at 4°C.
Upon warming to room temperature the slides will be put on Hyper film 3H for 5
days. The film will then be developed with D-19 developer.

 

Relative optical densities of the autoradiographic images will be measured
with an MCID image analysis system and the mean diffusion distances of 125I
-beacon determined.

 

In vitro autoradiography:

 

Frozen coronal plane sections of brain at the hypothalamus level are
thaw-mounted onto glass slides. Sections are incubated for 90 min at 40C
with O.5nM I-125 labeled protein in incubation medium containing
50mM Tris-HC1, 0.2% BSA, O.1mM phenylmethylsulfonyl fluoride, and 0.1 mM

 

 

phenatroline at pH 7.5. Non specific binding was determined by
incubation of adjacent sections in the presence of O.5uM unlabeled protein
added to the labeled protein in the incubation medium. After two 5 min washes
with cold incubation medium and one rinse in distilled water, the sections were
exposed together with I-125  labeled
plastic standards (Amersham) to Hyperfilm (Amersham) for 2 weeks.

 

2.                           Is
beacon a circulating protein?

 

For these studies, it is essential that a specific beacon antibody is
available. Currently the antibody is being produced in a collaborative contract
with Groupe Lipha.

 

Blood samples from lean and obese Israeli Sand rats will be collected
in the fed state and following a 24-hour fast. These blood samples will be
screened for binding to beacon antibody.

 

It is anticipated that these studies will take two months.

 

In Vitro Studies

 

1.                           Cellular
localization of beacon

 

Tissues including hypothalamus, liver, muscle, adipose tissue and
pancreas from lean and obese animals will be collected for these studies.

 

In situ hybridization:

 

Beacon is cloned into the Bluescript plasmid that allows for the
generation of both riboprobes (antisense and sense). For generation of sense
and antisense 35S-labeled riboprobes, the plasmids are linearized
and then subjected to in vitro transcription with T7 or T3 polymerase according
to the manufacturer’s instructions (Stratagene).

 

Tissue cryostat sections are mounted onto poly(L-lysine)-coated slides
and stored at - 70°C in dessicated boxes. After fixation in 4%
paraformaldehyde in 0.1M phosphate buffer, and acetylation in 0.1M triethanolamine/0.25% acetic anhydride for 10
min, hybridization is performed.  35S
-labeled riboprobes are used at concentrations of 1.5 - 2x107 cpm/m1.
Riboprobes are added to a hybridization solution of 50% formamide, 10mM TrisHCI
(pH 8), 0.05% transfer RNA, 10mM dithiothreitol, 10% dextran sulfate, 0.3M
NaCI, 1mM EDTA (pH 8) and 1 x Denhardt’s solution. Hybridization solution and a
glass coverslip are applied to each slide and slides are incubated o/n at 55-60°C.
After hybridization, coverslips are removed and the sections treated with RNase A, desalted, with a final high stringency
wash in 0.1x standard saline citrate at 6O°C.

 

Finally, the sections are dehydrated before being exposed to film
autoradiography or coated with LM-1 autoradiographic emulsion (Amersham).

 

 

Immunohistochemistry:

 

The beacon antiserum is a rabbit anti-beacon antiserum that binds to
the epitope that corresponds to beacon amino acids 1-33. Sections are prepared
as for in situ hybridization. Non-specific binding is blocked by incubating the
sections in normal serum. Tissue sections are incubated overnight at 4°C
with primary antiserum directed against beacon (1-33). Sections are washed with                                                PBS
buffer and then incubated with a biotinylated secondary antibody followed by an
avidin:biotinylated peroxidase complex (ABC) according to manufacturer’s
instructions (Vectastain Elite ABC kit, Vector Laboratories). The tissue
antigen is localized by incubation with a substrate for the enzyme. Colored end
product is visualised with Sigma Fast DAB peroxidase substrate kit. Sections
are counterstained with either neutral red or toluidine blue. Specificity of
the immunoreaction is confirmed by (i) omission of the primary antibody
and (ii) pre-absorption of the beacon antibody with synthetic beacon (1-33).

 

2.                           Signal
transduction pathways and beacon action

 

In these studies, both in vitro and in vivo experiments will be
performed.

 

In vitro experiments

 

RNA will be extracted from beacon-treated 3T3 cells in vitro and then a
number of commercially available microarrays will be used to determine the
effect of beacon treatment on gene expression.

 

Cells will be exposed to beacon via two methods:

 

(i)                                                 addition
of beacon to culture medium

 

(ii)                                              intracellular
introduction -liposome transfection/cell permeability/microinjection.

 

From a single experiment it is possible to get quantitative gene
expression data for up to 10,000 gene sequences.

 

In vivo experiments

 

Tissues will be collected from lean and obese Israeli Sand rats as well
as animals treated intracerebroventricularly with various beacon doses.

 

Initially, RNA will be extracted from the hypothalamus of 4-6 Israeli
Sand rats in each of the groups described above. These RNA samples will be
pooled and then hybridized to Clontech neurotransmitter specific microarrays.

 

These experiments will identify neurotransmitters and known signal
transduration genes that are up- or down-regulated following beacon treatment.
Following identification of key genes, ‘real-time’ PCR will be used to
accurately quantitate gene expression after beacon treatment.

 

These studies will identify signal transduction pathways involved in
beacon action. It is anticipated that the studies will be completed in six
months.

 

3.                           Beacon
binding to know receptors

 

These studies are being coordinated by Groupe Lipha. under supervision
of Dr Guy Augert. Studies examining whether beacon binds to approximately 20
known receptors involved in energy balance have been contracted to an
independent research company.

 

 

4.                           Identification
of beacon receptors

 

These experiments will be aimed at discovery and localization of beacon
receptors. As preliminary experiments examining beacon treatment in Israeli
Sand rats produced significant physiological effects, we anticipate beacon is
acting via a specific cellular receptor.

 

In addition, cDNA expression libraries will be prepared from Israeli
Sand rat tissue samples in three months for approximately $15,000.

 

In an attempt to identify the beacon receptor, we will utilize three
approaches:

 

(i) Surface Receptor Panning (method modified from Guo-Xi et al. (1992))

 

These studies will require a. beacon antibody and beacon fusion
protein. cDNA expression libraries transfected into target cells will be
screened for the production of cell surface expressed proteins capable of binding
to immobilized. Beacon will either be immobilized or tagged with a reporter
gene to aid detection of the interaction.

 

Immobilization of Anti-beacon

 

Petri dishes (60 mm diameter) will be coated with anti-beacon (301.μg
per dish in 3 ml of 50 mM Tris-HCI buffer, pH 9.5) for 2 h then washed three
times with 0.15 M NaCI. Phosphate buffered saline (3 ml PBS containing 0.1 %
bovine serum albumin (BSA)) will be added and the dishes incubated at 4°C
overnight.

 

Transfection and Panning

 

cDNA from the commercially produced cDNA expression library will be
transfected into COS cells. Exponentially grown cells will be plated (5x105
per 10 cm dish) in Dulbecco’s modified Eagles medium (DMEM) with 10% fetal calf
serum (FCS). After 24 h in a 37°C (5% C02/95% air) incubator cells
will be washed twice with Iscove’s modified Dulbecco’s medium (IMDM) which
contains L-glutamine and 25mM Hepes (pH 7.4.). Plasmid DNA (10μg), DEAE-dextran
and chloroquine (1001μM) will be mixed in 4 ml of IMDM, added to the dish and
returned to the incubator for 5 h. The cells will be washed with IMDM and then
with DMEM containing 5% FCS. The cells will be cultured in 10 ml DMEM with
10%  FCS for 3 days in the incubator for
maximal expression. Transfected cells will be detached with PBS containing 0.5
mM EDTA and .0.02% NaN3. After washing the cells twice with Krebs-Hepes buffer (KHB)
cells will he suspended in 1 ml KHB containing
0.5 M EDTA, 0.1% BSA and beacon and incubated 1.5 h at room temperature and
then 30 min on ice. After three washes to remove free beacon, cells will be
resuspended in 1 ml PBS containing 0.5 mM EDTA and 0.1 % BSA and plated on
antibody coated dishes. After 2 hours at room temperature, the dishes will be
washed gently three times with 3 ml PBS. Then cells remaining on the dishes
will be lysed and plasmid DNA recovered by the HIRT method and amplified in E.coli. The resulting transformants will be reintroduced
into COS cells by electroporation. The expression-panning-rescue cycle will be
repeated twice after which individual E.coli colonies
will be picked and plasmid DNA prepared by alkaline lysis miniprep procedure.

 

Beacon Binding Assay

 

Transfected cells will be incubated in KHB (106 cells per
m1) containing 125I-beacon and varying concentrations of unlabelled
beacon in a total volume of 1 ml for 45 min at room temperature. Incubations
were terminated by the oil floatation method and specific binding of
radiolabelled beacon to the COS cells measured using a gamma counter.

 

 

Sequence Analysis

 

Cloned cDNA will be sequenced using the ABI Prism Big Dye Terminator
Cycle Sequencing Ready Reaction Kit (PE Applied Biosystems).

 

(ii)                              Yeast
Two-hybrid’ System

(using the ProQuest Two-Hybrid System available from Life Technologies)

 

•                  Clone
the beacon gene in frame with the GAL4 sequence which encodes the DNA binding
domain in the vector pDBLeu.

 

•                  Construct
a cDNA expression library in the GAL4 activation domain of the yeast vector
pPC86. Life Technologies have a range available including; human (brain and
foetal brain), mouse (embryo 8.5 or 10 days, liver, brain and lymph node),
C.elegans, HeLa cell and rat (liver and brain). A custom library (Pssamomys obesus) can be created for this system by Life
Technologies within 8-12 weeks.

 

•                  Transform
the yeast strain MaV203 with the pDBLeu-beacon plasmid construct.

 

•                  Test
for self activation of the pDBLeu-beacon fusion protein and determine the
concentration of 3-Amino-1,2,4-Triazole (3AT) required to titrate basal HIS3
expression levels. HIS3 encodes imidazole glycerol phosphate deydratase, an
enzyme involved in histidine biosynthesis. This enzyme can be specifically
inhibited in a dose-dependent manner by 3AT. To maximise sensitivity of the
HIS3 reporter gene, strain MaV203 expresses a basal level of HIS3. By
determining the threshold of resistance to 3AT and including that concentration
of 3AT in plates lacking histidine, even slight increases in HIS3 reporter call
be detected, enhancing the likelihood of detecting even weak protein:protein
interactions.

 

•                  Transform
MaV203 cells containing the pDBLeu-beacon construct with pC86 library using
antibiotic resistance (ampicillin and kanamycin) to select for cells that
contain both plasmids and induce the HIS3 reporter gene.

 

•                  Purify
cells containing candidate interacting proteins then patch isolated colonies
onto a masterplate

 

•                  Replica
plate from the master plate onto selective plates to determine whether the
three reporter genes are induced.

 

•                  For
cells inducing the reporter genes confirm that DB-beacon and AD-fusion protein
(from the cDNA library) interact when AD-fusion protein is retested with fresh
DB-beacon plasmid by either a retransformation assay or a version of plasmid
shuffling.

 

•                  Protein:protein
interactions detected need to then be confirmed by biological assays such as
DNA sequencing of the cDNA clone to determine if the interacting protein has
been previously identified. This fusion protein can also be expressed in E.coli
and coprecipitation experiments using antibodies raised against beacon or
monoclonal antibodies raised against the GAL4-AD domains (protein tags derived
from the expression vectors). Other methods independent of the ProQuest system
will also be required to determine if these proteins truly interact these would
include surface receptor panning and fusion protein fishing.

 

 

(iii)Fusion Protein Fishing

 

Fusion protein fishing:

 

A full length beacon sequence will be cloned into the bacterial
expression plasmid pGEX (Pharmacia Biotech). pGEX vectors allow for inducible,
high level intracellular expression of genes as fusions with Schistoma
japonicum glutathione S-transferase
(GST). Induced bacterial cultures expressing pGEX-beacon will be lysed by
sonication in 50mM Tris-HCl (pH 7.4) containing 1% Triton X-100, 1% Tween 20,
2mM EDTA, 0.2mM phenylmethylsulfonyl fluoride, and l0μg/ml aprotinin. Affinity
resins for the isolation of beacon binding proteins are prepared by
immobilizing GST-beacon onto glutathione-Sepharose 4B beads (Pharmacia).
Similar resins can be prepared using GST alone to act as a control for the specificity
of interaction. Affinity resins are incubated in the presence of brain lysates
(or other tissue lysates of interest) and after extensive washing, proteins
bound to the resins are released either by boiling in SDS sample buffer or by
elution with Tris-HCl (pH 7.4) containing 0.5% Triton X-100. Binding proteins
are then separated by SDS-PAGE and visualised by silver staining or Coomassie
Blue staining. Regions of the gel containing beacon binding proteins are
excised and the gel slices digested and purified by anion-exchange and reverse
phase HPLC prior to amino acid sequencing.

 

It is necessary to use three different approaches to beacon receptor
identification. This will increase our chances of success in the shortest
possible time and to cover al1 possibilities that the putative beacon receptor
may be intracellular, extracellular or a membrane anchored protein. We
anticipate that these studies will take approximately 12 months.

 

5.                           Human
beacon gene studies

 

These studies will have two approaches.

 

(i)                                              Sequencing
of human beacon gene

 

In these studies, a sample of human beacon genes collected from obese
and lean individuals will be sequenced for beacon gene mutations and
polymorphisms. This will operate as a pilot study for eventual identification of
sequence variations associated with obesity.

 

DNA will be extracted from blood samples obtained from appropriately
selected donors. Detailed phenotypic will be simultaneously collected. The
Beacon gene will be PCR amplified from the extracted genomic DNA using primers
identified to reproducibly generate clean DNA bands. The amplified fragments
will be checked for contaminating bands, cleaned and sent out for DNA
sequencing by the Australian Genome Research Facility by contract.

 

(ii)                                          Upstream sequencing

 

In this study, we will attempt to sequence approximately 5 kb upstream
of the human beacon gene. This will allow study of the transcriptional control
elements that make up the human beacon gene promoter region. Analysis of this
region may identify variations that could be associated with the obese
phenotype through aberrant beacon gene expression control. Knowledge of these
elements may also provide a basis on which to externally control beacon gene
expression (described below in part iii). Before these studies begin, a genomic
library will be constructed in collaboration with a commercial company. This is
anticipated to take approximately two months. The upstream sequencing will then
follow.

 

Method

 

Standard methods of hybridisation will be employed to identify large
insert genomic clones (eg, cosmid, BAC, PAC) from a library containing the
Beacon gene. The library will either be generated by ourselves following
standard procedures or obtained from commercial sources.

 

 

Once identified the Beacon insert will be sub - cloned to a suitable
size for direct DNA Sequencing. Approximately 5 kb upstream of the Beacon gene
initiation codon will be sequenced to identify the primary components of the
promoter. The sequence will be analysed and compared to other mammalian gene
promoters for known transcription factor binding sites. Additionally the
sequence will be evaluated for whether further DNA sequence determination would
be required to fully encompass the extremities of the promoter region. Such
regions lying a greater distance from the beacon coding region may include
enhancer elements.

 

(iii)                                      Beacon Promoter Studies

 

This section will analyse the promoter region discovered above in
part(ii). If the promoter region appears to contain defined elements that may
be suitable for external modulation (eg control through transcription factor
expression), then a decision will be made to continue with the project.

 

This project would then involve the development of a tissue culture
based beacon gene promoter reporter system that could be used for screening of
gene expression modulating compounds. Such a system may then be adopted by
Merck-Lipha for a large scale compound screening program.

 

Method

 

The promoter region will be subcloned into an appropriate vector system
that would be capable of controlling expression of an easily detected reporter
gene such as luciferase or green fluorescence protein (GFP). The system would
be tested in a transient transfection assay for operation using culture human
target cells. Should this prove successful, then stably transfected cell lines
will be established, characterised and cultivated that would then form the
basis of a large-scale screening indicator system.

 

Standard methods of recombinant DNA cloning will be used in addition to
tissue culture, transfection and transformation procedures.Exhibit 4.2(g)

 

“CONFIDENTIAL TREATMENT REQUESTED.
CONFIDENTIAL PORTIONS OF THIS DOCUMENT HAVE BEEN OMITTED AND HAVE BEEN
SEPARATELY FILED WITH THE COMMISSION. 
CONFIDENTIAL TREATMENT HAS BEEN REQUESTED WITH RESPECT TO THE OMITTED PORTIONS.”

 

RESEARCH AND DEVELOPMENT OPTION AGREEMENT

 

	
  Made on

  	
   

  	
  30  August 2004

  
	
   

  	
   

  	
   

  
	
   

  	
   

  	
   

  
	
  BETWEEN

  	
   

  	
  AUTOGEN RESEARCH PTY LTD
  ACN 074 636 847 of Pigdons Road,
  Waurn Ponds, in the State of Victoria, Australia 

  
	
   

  	
   

  	
  (‘Chemgenex’)

  
	
   

  	
   

  	
   

  
	
  AND

  	
   

  	
  VERNALIS (R & D) LIMITED of Oakdene Court, 613 Reading Road, Winnersh, United Kingdom 

  
	
   

  	
   

  	
  (‘Vernalis’)

  

 

BACKGROUND

 

A.           Vernalis is a
British biotechnology company with strengths across the drug discovery and
development value chain. In addition to its successful development and
marketing of frovatriptan, the company maintains research programmes exploiting
its structure based drug discovery and CNS biology capabilities, investigating
the potential of a variety of novel molecules and targets in the fields of
oncology, obesity, inflammation and depression.

 

B.            Vernalis has agreed
to partner, in accordance with the terms of this research and development
option agreement, the Chemgenex research programme for the discovery and
validation of novel genes and proteins involved in depression and anxiety, the
product of these genes being able to be used as therapeutic agents or as a
target for drug discovery.

 

C.            It is clearly
understood that should Vernalis wish to expand this partnership beyond the
terms of this research and development option agreement the parties will
negotiate in good faith to develop an appropriate license and collaboration
agreement to cover an expanded research partnership.

 

OPERATIVE PROVISIONS

 

1.             DEFINITIONS

 

In this Agreement, except to the extent the
context otherwise requires:

 

‘Agreement’
means this agreement and any and all schedules as may be varied from time to
time in accordance with this agreement.

 

‘Affiliate’ means any company, partnership or other business entity which
Controls, is Controlled by or is under common Control with, either Party.

 

‘Chemgenex
IP’ means background Intellectual Property Rights and Know How and
Materials pertaining to the Licensed Field that Chemgenex has developed or
which it owns or is licensed to use prior to the Commencement Date;

 

 

‘Commencement Date’ means the date on which the Services will
commence, as specified in Schedule 1;

 

‘Competent Authority’ means any national or local
agency, authority, department, inspectorate, minister, ministry official,
parliament or public or statutory person, (whether autonomous or not) of any
government of any country having jurisdiction over any activities contemplated
by this Agreement or Parties.

 

‘Confidential Information’ in the
case of obligations on Vernalis Confidential Information shall mean Chemgenex
IP, in the case of obligations on Chemgenex Confidential Information shall mean
Vernalis IP and Vernalis Materials and in the case of both Vernalis and
Chemgenex Confidential Information shall mean Results, Programme Intellectual
Property and all trade secrets and confidential information relating to the
business affairs or finances of the other Party supplied or otherwise made
available or coming into possession of the other party in relation to the
performance of this Agreement and the terms of this Agreement.

 

‘Control’ means the ownership either
directly or indirectly of more than 50% of the issued share capital or any
comparable equity or ownership interest with respect to a business entity or
the legal power to direct or cause the direction of the general management and
policies of the Party in question.

 

‘Documents’ means reports, notes, charts,
computations, analyses, recordings, books, tapes, discs, computer programmes,
samples of material, written data and any other media on which Know How can be
permanently stored.

 

‘Extended
Term’ means 6 months commencing on the conclusion of the Initial
Term;

 

‘Force
Majeure’ means a circumstance beyond the reasonable control of the
Parties which results in a Party being unable to observe or perform on time an
obligation under this Agreement.  Such
circumstances shall include but shall not be limited to:

 

(a)           acts of God, lightning
strikes, earthquakes, floods, storms, explosions, fires and any natural
disaster;

 

(b)           acts of war, acts of public
enemies, terrorism, riots, civil commotion, malicious damage, sabotage and
revolution; and

 

(c)           strikes;

 

‘Know How’ means all technical and other information which is not in the
public domain, including information comprising or relating to concepts,
discoveries, data, designs, formulae, ideas, inventions, methods, models,
assays, plans, procedures, designs for experiments and tests and results of
experimentation and testing (including results of research or development),
processes (including manufacturing processes, specifications and techniques),
laboratory records, chemical, pharmacological, toxicological, clinical,
analytical and quality control data, trial data, data analyses, reports,
manufacturing data or summaries and information contained in submissions to and
information from ethical committees and regulatory authorities.  Know How includes Documents containing Know
How, including but not limited to any rights including trade secrets,
copyright, database or design rights protecting such Know How.  The fact that an item is known to the public
shall not be taken to preclude the possibility that a compilation including the
item, and/or a development relating to the item, is not known to the public.

 

‘Initial
Term’ means the period 6 months from the Commencement Date;

 

‘Intellectual
Property Rights’ includes:

 

(a)           any rights in or relating
to an invention;

 

(b)           copyright, trade mark,
design, patent, semiconductor,  circuit
layout and/or plant breeders’ rights;

 

(c)           trade, business, company
and/or domain names;

 

 

(d)           any right to have
Confidential Information kept confidential;

 

(e)           Know How and/or

 

(f)            other similar proprietary
rights,

 

or any rights to registration of such rights
existing anywhere in the world, whether created before, on or after the date of
this Agreement.

 

‘Legal Requirement’ means any present or future law, regulation, directive,
instruction, direction or rule of any Competent Authority from time to
time in force.

 

‘Licensed
Field’ means the field of human therapeutic applications for the
treatment or prevention of depression and/or anxiety disorders including
without limitation obsessive compulsive disorders and any other disorders in
which SSRI (selective serotonin reuptake inhibitors) are effective.

 

‘License and
Collaboration Agreement’ means a future contract pursuant to which
Vernalis has the exclusive right to develop, use and otherwise exploit the
Results and the Programme Intellectual Property beyond the Extended Term;

 

‘Materials’
means any biological or chemical substance including any;

 

(a)           organic or inorganic element;

 

(b)           nucleotide or nucleotide
sequence including DNA and RNA sequences;

 

(c)           gene;

 

(d)           protein including any peptide or
amino acid sequence, enzyme, antibody or protein conferring targeting
properties and any fragment of a protein or a peptide enzyme or antibody;

 

(e)           drug or pro-drug;

 

(f)            assay or reagent;

 

(g)           data for the deivation of
molecular structures including NMR spectra, X Ray diffraction patterns and
other primary experimental information assignments and other calculations
required for determination of the structure, and co-ordinates of the derived
molecular structure;

 

(h)           computer programmes or
algorithms.

 

‘Option’ means the right granted to Vernalis to exclusively negotiate the
Licence and Collaboration Agreement under clause 3.1.

 

‘Party’
means either Chemgenex or Vernalis as the context dictates;

 

‘Payment #1’
means the GB£[*] payment to be made by Vernalis to Chemgenex pursuant to clause
4.1;

 

‘Payment #2’
means the  GB£[*] payment to be made by
Vernalis to Chemgenex pursuant to clause 4.2;

 

‘Programme’
means the programme described at Schedule 2;

 

‘Programme Intellectual
Property’ means any and all Intellectual Property Rights subsisting
in the Results

 

‘Results’
means all Know How, Materials and Documents created, resulting from or arising
in the course of carrying out the Programme or the Services.

 

‘Schedule’
means a Schedule to this Agreement;

 

‘Services’
means the research and development services to be provided by or on behalf of
Chemgenex as specified in Schedule 2 as may be amended from time to time
pursuant to this Agreement

 

2

 

‘Vernalis
IP’ means background Intellectual Property Rights and Know How
pertaining to the Licensed Field that Vernalis has developed prior to the
Commencement Date and all Vernalis Materials;

 

‘Vernalis Materials’ means those Materials owned by Vernalis that may be supplied to
Chemgenex under this Agreement.

 

2.             SERVICES

 

2.1           Chemgenex shall provide the
Services exclusively to Vernalis for the Initial Term and for the Extended
Term, subject to the provisions of clause 2.2, on the terms of this
Agreement.  For the avoidance of doubt
Chemgenex shall in provision of the Services achieve those milestones set out
in Schedule 2, as such milestones may be varied or amended in accordance
with the decisions of the Research Committee, in respect of the relevant term.

 

2.2           Vernalis may require and
Chemgenex shall be obliged to provide the Services for the Extended Term on the
provision by Vernalis of written notice to Chemgenex no later than 7 days prior
to expiry of the Initial Term that it wishes to extend the provision of
Services pursuant to this clause of the Agreement.  In the event that Vernalis does not exercise
its right to extend the provision of the Services for the Extended Term, the
provisions of Clause 9.4 shall apply.

 

2.3           Chemgenex shall perform the
Services during the Initial Term, and if relevant, the Extended Term;

 

(a)           in a good scientific manner and
with reasonable skill and diligence;

 

(b)           in accordance with all relevant
Legal Requirements and shall be responsible for obtaining all necessary
approvals therefore from any Regulatory Authority or applicable Competent
Authority

 

(c)           keep or cause to be kept
detailed written laboratory notebooks and other records and reports of the
Results and progress of the Programme and the Services (including in sufficient
detail for patent purposes);

 

(d)           but otherwise, shall exercise
its independent discretion as to the most appropriate and effective manner of
providing the Services.

 

2.4           In the event that Vernalis
agrees to provide to Chemgenex any Vernalis Materials, Chemgenex agrees that
such materials will be provided under terms of a separate material transfer
agreement with Vernalis.

 

2.5           Chemgenex agrees to promptly
disclose to Vernalis and provide Vernalis with access to all Results and
Programme Intellectual Property generated during or arising out of the
Programme and the Services and Vernalis shall have the right to make copies of
all such Results and Programme Intellectual Property as it may require.

 

3.             OPTION

 

3.1           Chemgenex hereby grants to
Vernalis an irrevocable exclusive option (“Option”) to negotiate in good faith
a Licence and Collaboration Agreement which will govern the terms of the future
relationship of the Parties and will include terms relating to, but not limited
to:

 

(a)           the grant to Vernalis of an
exclusive sub-licensable world-wide licence under Chemgenex’s interest in the
Programme Intellectual Property and Results and Chemgenex IP to develop, use
and otherwise exploit and commercialise the Programme Intellectual Property and
the Results;

 

(b)           timing and quantum of milestone
payments;

 

(c)           royalty share on products;

 

3

 

(d)           IP protection costs and rights;
and

 

(e)           Pro forma licence agreements

 

3.2           Vernalis may exercise the Option
from the date of expiry of the Extended Term and for a period of [*]
days thereafter (“The Option Period”). 
For the avoidance of doubt, the Option shall not be exercisable by
Vernalis if Vernalis has not exercised its right to extend the provision of
Services under this Agreement for the Extended Term.

 

3.3           In consideration of the grant of
the Option, Vernalis shall pay to Chemgenex [*] pounds (£[*])
on the date of exercise of the Option and a further [*] pounds (£[*])
on the date of execution of the License and Collaboration Agreement.

 

3.4           In the event;

 

3.4.1        that the Parties acting
reasonably and with good faith have not executed a Licence and Collaboration
Agreement within eighteen (18) months of the end of the Extended Term; or;

 

3.4.2        Vernalis does not exercise its
Option during the Option Period;

 

the Parties shall negotiate in good faith the
terms of a licence to Chemgenex of Vernalis’ joint interest in the Results and
Programme Intellectual Property.  For the
avoidance of doubt, unless and until the Parties agree on the definitive terms
of such licence, Chemgenex shall have no right to use, licence or otherwise
exploit in any way the Results or the Programme Intellectual Property without
Vernalis’ prior written consent except to the extent required to perform the
Services in accordance with this Agreement.

 

4.             PAYMENT

 

4.1           Vernalis shall make Payment #1
to Chemgenex within 30 days of the date of an invoice from Chemgenex raised on
or after the Commencement Date.

 

4.2           If Vernalis exercises its right
to extend the provision of Services under this Agreement for the Exteneded Term
under Clause 2 it must make Payment #2 within 30 days of the date of an invoice
from Chemgenex raised on or after 32 weeks of the Commencement Date.

 

4.3           The Payments are exclusive of
taxes, duties and charges imposed or levied in Australia or overseas in
connection with the Services.  Without
limiting the foregoing, Vernalis shall be liable for any new taxes, duties or
charges imposed subsequent to the Commencement Date in respect of this
Agreement.   Chemgenex shall provide to
Vernalis all such assistance requested by Vernalis in reclaiming, minimising or
avoiding payment of any such taxes, duties or charges paid or payable by
Vernalis.

 

5.             LAWFUL DIRECTIONS

 

5.1           In the discharge of its duties
Chemgenex shall comply with all reasonable resolutions, regulations and
directions of Vernalis as may lawfully be given from time to time as to the
nature and scope of the Services to be provided.

 

5.2           Nothing in clause 5.1 shall
affect Chemgenex’s right to exercise its own judgement and to
utilise its skills as it considers most appropriate in order to achieve
compliance with the said resolutions, regulations and directions or otherwise
to comply with its obligations under this Agreement.

 

4

 

6.             RESEARCH COMMITTEE

 

6.1           The Parties agree to establish
not later than 14 days after the Commencement Date, a joint research committee
(‘Research Committee’) to oversee the progress of the Programme and Services.

 

6.2           The Research Committee will be
comprised of the following participants:

 

(a)           two representatives of
Chemgenex, being the Chief Executive Officer and the Senior Director of
Research and

 

(b)           two representatives of Vernalis,
being Director of Research and a senior Vernalis scientist.

 

Each Party will use reasonable endeavours to
achieve continuity of representation on the Research Committee, but shall be
entitled, on prior written notice to the other to replace its representative
with a person acceptable to the other Party acting reasonably.  The quorum for meetings of the Research
Committee shall be 2 representatives provided that there is one representative
from each of Vernalis and Chemgenex.

 

6.3           The purpose of the Research
Committee will be to monitor the Programme, discuss and agree the nature and
scope of the Services, and to make decisions in relation to the patenting of
inventions forming part of the Results. 
Except as regards variations or additions to the Services, the Research
Committee shall have no power to vary this Agreement except in accordance with
Clause 19.

 

6.4           The Research Committee will meet
on a monthly basis by videoconference and on a quarterly basis by face-to-face
meetings, unless otherwise agreed between the Parties.  Face to face meetings will alternate between
the Parties respective premises and each Party shall be responsible for all of
its own expenses of attending and participating in the Research Committee.

 

6.5           Minutes of each meeting shall be
prepared and issued within thirty (30) days thereafter and responsibility for
preparing such minutes shall alternate between the Party’s
representatives.  Minutes shall not be
deemed to be finalised until the Party’s representative who did not prepare
such minutes reviews and confirms the accuracy of the minute in writing.  Chemgenex will produce a quarterly report
detailing Results and progress of the Services to be issued within 14 days
prior to each of the aforementioned quarterly face-to-face meetings.

 

6.6           If unanimous agreement cannot be
reached by the Research Committee within ten (10) days, either Party may,
on giving notice to the other, refer the matter in dispute to the Chief
Executive Officers of each of the Parties who shall work in good faith to try
to resolve such dispute.

 

7.             CONSULTANT’S STATUS

 

7.1           Chemgenex is an independent
contractor without authority to bind Vernalis by contract or otherwise and
neither Chemgenex nor Chemgenex’s personnel are agents or employees of Vernalis
by virtue of this Agreement.

 

7.2           Chemgenex acknowledges it has
sole responsibility in relation to and shall indemnify Vernalis in respect of
any and all payment, if any, of superannuation, workers’ compensation and taxes
incidental to employment in respect of its own personnel. Chemgenex further
acknowledges that neither it nor its personnel have, pursuant to this
Agreement, any entitlement from Vernalis in relation to any form of employment
or related benefit.

 

8.             CONFIDENTIALITY

 

8.1           Each Party (the receiving Party)
undertakes and agrees to:

 

8.1.1        only use the Confidential
Information which comes into its possession pursuant to this Agreement for the
purposes envisaged under this Agreement and not to use the same for any other
purpose whatsoever;

 

5

 

8.1.2        ensure that only those of its
officers employees and consultants who are directly concerned with the carrying
out of this Agreement have access to the Confidential Information on a strictly
applied “need to know” basis; are informed of the secret and confidential
nature of it and are bound by confidential obligations no less strict than
those provided herein;

 

8.1.3        keep the Confidential
Information secret and confidential and not directly or indirectly to disclose
or permit to be disclosed, make available or permit to be made available the
same to any third party for any reason without the prior written consent of the
disclosing party;

 

Affiliates shall not
be regarded as third parties so long as they are bound by confidentiality
obligations which are not less strict than those set forth herein.

 

8.2           The disclosing Party shall
clearly identify the Confidential Information as confidential except for the
Results and Programme Intellectual Property which shall automatically be deemed
to be identified as Confidential Information.

 

8.3           The obligations of confidence
referred to in Clause 8.1 shall not extend to any Confidential Information
which:

 

8.3.1        is or becomes generally
available to the public otherwise than by reason of breach by the receiving
Party of the provisions of this Clause;

 

8.3.2        is known to the receiving Party
and is at its free disposal prior to its receipt from disclosing Party, or in
the case of Results prior to creation of such Results, or was generated
independently by the receiving Party or a third party in circumstances where it
has not be derived directly or indirectly from the Confidential Information
provided that evidence of such knowledge or independent generation is furnished
by the receiving Party to the disclosing Party within thirty (30) days of
receipt of that Confidential Information; or

 

8.3.3        is subsequently disclosed to the
receiving Party without obligations of confidence by a third party owing no
such obligations to the disclosing Party in respect of that Confidential
Information;

 

8.3.4        is required by law to be
disclosed (including as part of any regulatory submission or approval process)
and then only when prompt written notice of this requirement has been given to
the disclosing Party so that it may, if so advised, seek appropriate relief to
prevent such disclosure provided always that in such circumstances such
disclosure shall be only to the extent so required and shall be subject to
prior consultation with the disclosing Party with a view to agreeing timing and
content of such disclosure;

 

8.4           All Confidential Information of
a Party shall remain the property of that Party.  In the event that a court or Competent
Authority assumes partial or complete control over the assets of a receiving
Party based on the insolvency or bankruptcy of that Party, the receiving Party
shall;

 

8.4.1        promptly notify such court or
Competent Authority;

 

(a)           that Confidential Information received from or otherwise belonging
to the disclosing Party under this Agreement remains the property of the
disclosing Party; and

 

(a)          of the confidentiality
obligations under this Agreement; and

 

8.4.2        to the extent permitted by law,
take all steps necessary or desirable to maintain the confidentiality and
security of the disclosing Party’s Confidential Information and to ensure that
the court or Competent Authority maintains that Confidential Information in
confidence in accordance with this Agreement.

 

8.5           The obligations of the Parties
under Clause 8.1 to 8.4 shall survive the expiration or termination of this
Agreement for whatever reason for a period of ten (10) years.

 

6

 

8.6           Neither Party shall be permitted
to publish the Results or the Programme Intellectual Property without the prior
written consent of the other.

 

9.             INTELLECTUAL
PROPERTY AND COMMERCIALIZATION

 

9.1           Chemgenex owns all Chemgenex IP,
and licenses this to Vernalis for the term of this Agreement.

 

9.2           Vernalis owns all Vernalis IP,
and may choose to license parts of this to Chemgenex for the term of this
Agreement.

 

9.3           Subject to the provisions of
Clause 9.4 only, the Parties will jointly own in equal undivided shares all
Programme Intellectual Property and any modifications or improvements
discovered or made to Programme Intellectual Property and neither Party shall
have the right to themselves use or to licence to a third party the right to
use the Programme Intellectual Property except for the Programme and Services
or in the case of Vernalis to carry out internal research in the Field, or if
executed, in accordance with the terms of the Licence and Collaboration
Agreement, and neither Party shall during the period of this Agreement assign,
transfer, mortgage, charge or otherwise dispose of or encumber Chemgenex IP,
Vernalis IP or Programme Intellectual Property, except in accordance with the
terms of this Agreement.

 

9.4           [*]

 

9.5           Chemgenex shall promptly disclose to Vernalis all Results and Programme
Intellectual Property..  Each party may
propose to the other Party that a filing for patent rights claiming an
invention be made.  Provided that the
Parties have agreed that such filing be made and have approved the content of
such filing all costs and expenses associated with such patent rights incurred
shall be shared equally between the Parties and the Party responsible for such
filing shall provide the other Party with a quarterly invoice for fifty percent
of the costs and expenses incurred by the responsible party during such
quarter.  The Party responsible for the
filing prosecution and maintenance of such patent rights shall provide copies
of all material correspondence with the respective patent office to the other
Party and shall use reasonable efforts to keep the other Party informed of the
progress of such prosecution.  No filing
for or patent rights shall be abandoned by the responsible party unless both
Parties have agreed to such action after consultation with their respective
patent counsel.  In the event the Parties
fail to agree upon a filing for such patent right or the continued prosecution
and maitantenance of a filed patent right, the party desiring to file,
prosecute and/or maintain patent rights shall be permitted to make such filing
or continue such prosecution at its own expense provided that in such event the
invention or patent right in question shall cease to form part of Programme
Intellectual Property and shall become owned by that Party.  The Party declining to contribute to the cost
of such filing, prosecution or maintenance or to approve the content of such
filing (as the case may be) shall do all such acts and things and shall execute
all such deeds and documents as are necessary to vest such rights in the other
Party.

 

9.6           In the event of an infringement of a patent right forming part of
Programme Intellectual Property, the Parties shall decide the best way for the
Parties to proceed.

 

10.           IMPLIED TERMS

 

10.1         Each Party represents and
warrants to the other Party that it has the legal power authority and right to
enter into this Agreement free from any conflicting right owed to a third party
and to perform its obligations hereunder

 

10.2         Subject to Clause 10.1, any condition of
warranty which would otherwise be implied in this Agreement is hereby excluded.

 

7

 

10.3         Where legislation implies in
this Agreement any condition or warranty, and that legislation avoids or
prohibits provisions in a contract excluding or modifying the application of or
exercise of or liability under such condition or warranty, the condition or
warranty shall be deemed to be included in this Agreement.  However, the liability
of Chemgenex for any breach of such condition or warranty shall be limited, at
the option of Chemgenex to the supply of services again or the payment of the
cost of having such service supplied again.

 

10.4         Vernalis
and Chemgenex shall not disclose any terms or conditions of this Agreement to
any third party without the prior consent of the other party, except as
required by applicable law, stock exchange requirements or local regulations or
to a third party with whom Chemgenex or Vernalis has entered into or proposes
to enter into a business relationship, provided that such third party shall
enter into a confidentiality agreement with, or otherwise owe a duty or
confidentiality to Vernalis or Chemgenex, as applicable.

 

10.5         Except
as required by law, order or regulation of a governmental agency,stock exchange
or a court of competent jurisdiction, no other announcement, public release or
notice of any kind may be issued without the express written consent of both
Parties, which consent shall not be unreasonably withheld, provided, however,
the Parties shall prepare a joint press release announcing the transaction set
forth in this Agreement, the content and nature of which is subject to prior
written agreement between the Parties, to be issued promptly upon execution of
this agreement.

 

11.           TERM AND TERMINATION

 

11.1         This Agreement shall commence on
the Commencement Date and shall expire at the end of the Initial Term unless
extended under the provisions of Clause 2 whereupon it will continue until the
earliest of execution of the Licence and Collaboration Agreement or conclusion
of a licence of Vernalis joint interest in Programme Intellectual Property
pursuant to Clause 3.3.

 

11.2         If the Agreement expires at the
end of the Initial Term, Vernalis shall not, for the avoidance of doubt, be
required to make Payment # 2.

 

11.3         Without limiting the generality
of any other clause in this Agreement, either Party (the ‘first Party’) may
terminate this Agreement immediately by notice in writing if:

 

(a)           the other Party (the ‘second
Party’) is in material breach of any term of this Agreement which in the case
of a breach capable of remedy is not remedied within thirty (30) days of it
being notified by of the breach the first Party;

 

(b)           the second Party becomes,
threatens or resolves to become or is in jeopardy of becoming subject to any
form of insolvency administration;

 

(c)           the second Party, being a
partnership, dissolves, threatens or resolves to dissolve or is in jeopardy of
dissolving;

 

11.4         Upon termination by Chemgenex
under Clause 11.3, Chemgenex shall have the right to continue Programme and
Vernalis hereby grants to Chemgenex an exclusive worldwide sub-licensable
licence to use and explicit Vernalis’ interest in the Programme Intellectual
Property subject to the Parties negotiation in good faith the consideration
payable to Vernalis for the grant of such licence

 

11.5         Upon termination by Vernalis
under Clause 11.3, Vernalis shall have the right to continue the Programme and
Chemgenex hereby grants to Vernalis an exclusive worldwide sub-licensable
licence to use and exploit the Chemgenex IP and Chemgenex’s interest in the
Programme Intellectual Property.  The
Parties shall negotiate in good faith the consideration payable to Chemgenex
for the grant of such licences.

 

11.6         If the Agreement expires
pursuant to Clause 11.2, the Parties agree that;

 

(a)           Vernalis shall deliver up any
Chemgenex property in its possession, custody or control;

 

(b)           Chemgenex shall retain any
moneys paid;

 

8

 

(c)           Chemgenex be regarded as
discharged from any further obligations under this Agreement; and

 

11.7         Upon termination by either Party
under Clause 11.3, the second Party (as defined in Clause 11.3) shall provide
or deliver all such assistance, information, Documents or Materials necessary
or desirable for effecting the licences set out in Clauses 11.4 and 11.5.

 

11.8         Termination of this Agreement
for whatsoever reason shall not affect the accorded rights of the Parties
arising in any way out of this agreement as at the date of termination and all provisions
which expressly or by implication survive termination of this Agreement shall
remain in full force and effect.

 

12.           FURTHER ASSURANCES

 

Each party must do all things and execute all
further documents necessary to give full effect to this Agreement and refrain
from doing anything that might hinder the performance of this Agreement.

 

13.           FORCE MAJEURE

 

13.1         Neither Party shall be liable
for any delay or failure to perform its obligations pursuant to this Agreement
if such delay is due to Force, Majeure.

 

13.2         If a delay or failure of a Party
to perform its obligations is caused or anticipated due to Force Majeure, the
performance of that Party’s obligations will be suspended.

 

13.3         If a delay or failure by a Party
to perform its obligations due to Force Majeure exceeds sixty (60) days, either
Party may immediately terminate the Agreement on providing notice in writing to
the other Party.

 

13.4         If this Agreement is terminated
pursuant to clause
13.3, Chemgenex shall refund moneys previously paid by Vernalis pursuant to
this Agreement for goods or services not provided by Chemgenex to Vernalis.

 

14.           SUB-CONTRACTS

 

Chemgenex shall not subcontract the
performance of any of its obligations other than to its existing collaboration
partners (Deakin University, the International Diabetes Institute and the
Southwest Foundation for Biomedical Research) without the prior written consent
of Vernalis.  In relation to any
authorised sub-contractor, Chemgenex shall ensure that the terms of its
agreements with such subcontractors impose restrictions and obligations which
are at least as restrictive as those contained in this Agreement (including
without limitation obligations of confidence and non-use) and retain or reserve
to Chemgenex all such rights as are necessary to fulfil its obligations to
Vernalis hereunder, including without limitation ownership of Intellectual
Property Rights.  Chemgenex further
agrees that it shall be responsible for all acts and omissions of such sub-contractor
as though they were part of Chemgenex.

 

15.           ENTIRE AGREEMENT

 

This Agreement constitutes the entire
agreement between the Parties and supersedes all prior representations,
agreements, statements and understandings, whether verbal or in writing.

 

16.           PRECEDENCE

 

16.1         The documents comprising this
Agreement shall be read in the following order of precedence:

 

(a)           the clauses of this Agreement;

 

(b)           the Schedules.

 

9

 

16.2         Where any conflict occurs
between the provisions contained in two or more of the documents forming this
Agreement, the document lower in the order of precedence shall where possible
be read down to resolve such conflict. 
If the conflict remains incapable of resolution by reading down, the
conflicting provisions shall be severed from the document lower in the order of
precedence without otherwise diminishing the enforceability of the remaining
provisions of that document.

 

17.           ASSIGNMENT AND
NOVATION

 

17.1         Neither Party shall without the
prior written consent of the other assign the benefit and/or burden of this
Agreement except that Vernalis may assign this Agreement to an Affiliate or to
any successor in connection with any merger, consolidation or sale or disposal
of all or substantially all of its assets and / or business, without the prior
consent of Chemgenex.

 

18.           WAIVER

 

18.1         No right under this Agreement
shall be deemed to be waived except by notice in writing signed by each Party.

 

18.2         A waiver made by either Party
pursuant by clause 18.1
will not
prejudice its rights in respect of any subsequent breach of the Agreement by
the other Party.

 

18.3         Subject to clause 18.1, any failure by
either Party to enforce any clause of this Agreement, or any forbearance, delay
or indulgence granted by either Party will not be construed as a
waiver
of that Party’s rights under this Agreement.

 

19.           VARIATION

 

The provisions of this Agreement shall not be
varied, except by agreement in writing signed by the Parties.

 

20.           SEVERABILITY

 

If any provision of this Agreement is held
invalid, unenforceable or illegal for any reason, the Agreement shall remain
otherwise in full force apart from such provisions which shall deemed deleted.

 

21.           SURVIVAL OF
AGREEMENT

 

21.1         Subject to any provision to the
contrary, this Agreement shall enure to the benefit of and be binding upon the
Parties and their successors, trustees, permitted assigns or receivers but
shall not enure to the benefit of any other persons.

 

21.2         The covenants, conditions and
provisions of this Agreement which are capable of having effect after the
expiration of the Agreement shall remain in full force and effect following the
expiration of the Agreement.

 

22.           GOVERNING LAW

 

This Agreement will be governed by and
construed according to the law of the jurisdiction specified in Schedule 1.  All disputes as set out in this Agreement
shall be subject to the jurisdiction of the High Court in London, England.

 

10

 

23.           NOTICES

 

23.1         Notices under this Agreement may
be delivered by hand, by mail or by facsimile to the addresses specified in Schedule 1.

 

23.2         Notice will be deemed given:

 

(a)           in the case of hand delivery,
upon written acknowledgment of receipt by an officer or other duly authorised
employee, agent or representative of the receiving Party;

 

(b)           in the case of posting, three
days after despatch; or

 

(c)           in the case of facsimile, upon
receipt of transmission if received on a business day or otherwise at the
commencement of the first business day following transmission.

 

24.           EXECUTION

 

This Agreement may be executed in counterparts by the respective Parties, each of which
when so executed shall be deemed to be an original and all of which
taken together shall constitute one and the same agreement, provided that this
Agreement shall be of no force and effect until the counterparts are exchanged.

 

25.           THIRD PARTY RIGHTS

 

No term of this Agreement shall be
enforceable under the Contract (Rights of Third Parties) Act 1999 by a person
not a party to this Agreement.

 

26.           INTERPRETATION

 

In this Agreement, unless the contrary
intention appears:

 

(a)           the clause headings are for
convenient reference only and have no effect in limiting or extending the
language of the provisions to which they refer;

 

(b)           a cross reference to a clause
number is a reference to its clauses;

 

(c)           words in the singular number
include the plural and vice versa;

 

(d)           words importing a gender include
any other gender;

 

(e)           a reference to a person includes
a partnership and a body, whether corporate or otherwise;

 

(f)            a reference to a clause is a
reference to a clause or subclause of this Agreement;

 

(g)           a reference to a clause is a
reference to a subclause of the clause in which that reference is made;

 

(h)           where a word or phrase is given a
particular
meaning, other parts of speech and grammatical forms of that word or phrase
have corresponding meanings;

 

(i)            a reference to a Schedule includes
a reference
to any part of that Schedule which is not physically annexed to this
Agreement but which is incorporated by reference;

 

(j)            the recitals to this Agreement
do not form part of the Agreement;

 

(k)           monetary references are
references to Australian currency; and

 

(l)            ‘includes’ is not a term of
limitation.

 

11

 

SCHEDULE 1

Contract details

 

	
  Commencement
  Date:

  	
   

  	
  1
  September 2004

  
	
   

  	
   

  	
   

  
	
  Governing
  Law:

  	
   

  	
  The law of
  England and the jurisdiction of the English Courts.

  
	
   

  	
   

  	
   

  
	
  Notices:

  	
   

  	
  Chemgenex

  
	
   

  	
   

  	
  Dr Greg Collier

  CEO and Managing Director

  PO Box 1069, Grovedale, Victoria 3216, AUSTRALIA

  Fax: +61 3 5227 1322

  
	
   

  	
   

  	
   

  
	
   

  	
   

  	
  Vernalis

  Dr. Mike Wood

  Director of Research

  Vernalis

  Oakdene Court, 613 Reading Road, Winnersh, RG41 5UA, UK

  Fax: +44 (0)118 989 9300

  

 

12

 

SCHEDULE 2

Programme & Services

 

The following research plan for the Initial
Term and the Extended Term is proposed, but it is the clear understanding of
the Parties that the programme is dynamic and may be redirected by the Research
Committee to maximise the value of the programme.

 

[*]

 

EXECUTED as an agreement in England.

 

13

 

	
  SIGNED on behalf of 

  	
  )

  	
   

  
	
  AUTOGEN RESEARCH PTY LTD

  ACN 074 636 847 by an Authorised Officer

  in the presence of:

  	
  )

  )

  )

  	
   

  
	
   

  	
  )

  	
   

  	
   

  
	
   

  	
   

  	
  Ù

  	
  Signature of Authorised
  Officer

  
	
   

  	
   

  	
   

  
	
   

  	
   

  	
   

  	
   

  
	
  Ù

  	
  Signature of witness

  	
   

  	
   

  
	
   

  	
   

  	
   

  	
   

  
	
   

  	
   

  	
   

  	
   

  
	
  Ù

  	
  Name of witness (print)

  	
   

  	
   

  
	
   

  	
   

  	
   

  	
   

  
	
   

  	
   

  	
   

  	
   

  
	
  SIGNED by 

  	
  )

  	
   

  
	
  VERNALIS (R & D) LIMITED by an
  Authorised

  Officer in the presence of:

  	
  )

  )

  )

  	
   

  
	
   

  	
  )

  	
   

  	
   

  
	
   

  	
   

  	
  Ù

  	
  Signature of Authorised
  Officer

  
	
   

  	
   

  	
   

  
	
   

  	
   

  	
   

  	
   

  
	
  Ù

  	
  Signature of witness

  	
   

  	
   

  
	
   

  	
   

  	
   

  	
   

  
	
   

  	
   

  	
   

  	
   

  
	
  Ù

  	
  Name of witness (print)

  	
   

  	
   

  

 

14

Source: [{"source": "alea-institute/alea-institute/kl3m-data-edgar-agreements/train-00086-of-00352.parquet"}, [{"source": "alea-institute/alea-institute/kl3m-data-edgar-agreements/train-00086-of-00352.parquet"}]]