Note: This is the text of an FDA report on Searle

EIR 4/25/77 to 8/4/77                      Searle Laboratories
JSA/DME/JT/LF                              Div. G.D. Searle & Co.
                                           4901 Searle Parkway
                                           Skokie, Illinois 60076

SUMMARY OF FINDINGS

Authentication of this study was performed primarily by com-
paring available raw data with the submission to FDA.  This
was a problem, at times, due to the lack of some data and
difficulty in locating other material.  The majority of
material relating to Aspartame was already under FDA  seal
at Searle.  However, during this investigation we discovered
various documents and notebooks that were not.

In some cases original data could be recorded in several areas,
making it difficult, and sometimes impossible to determine
which was actually the original.  This was a particular problem
in dealing with dates of deaths, as some conflicted on the
"source" documents.  Many of the responsible individuals invol-
ved with the study, including stability testing of DKP, are no
longer employed by Searle.  Dr. K.S. Rao, Study Monitor,
the only individual who could have possible answered some quest-
tions, had left Searle.  He was contacted, but permission for
an interview was refused by his attorney.  Due to the absence
of various individuals it was not always possible to a accurately
determine methods used in some analyses and operations carried
out in conducting this study.  In a number of areas, including
chemistry, statistics, diet preparation and feeding, it was
necessary to use assumptions, or information supplied by current
employees who were not involved with the study.

At the beginning of this investigation , Mr. James R. Phelps,
Vice-President and General Counsel for G.D. Searle & Co., advised
us that an attorney and scientific coordinator would have to be
present at all times to protect their interest in the data.
This did not present any insurmountable problems, but on several
occasions an attorney would question our request for data,
stating that it was not relevant for authentication.  At no time
did we make any statement to the effect that our goal was to authen-
ticate the study.  Two memos were discovered dealing with reaction
of animals to the diet.  This was a significant factor in the
study.  Permission to copy them was initially refused, but
finally granted after Searle was contacted by FDA General Coun-
sel.  We were not allowed to make xerox copies of any documents
for about two and one-half weeks, due to Searle's concern over
confidentiality.  This was eventually reconciled between Searle
and FDA General Counsel.

                           (1)

The major discrepancies concerning Study PD 988S73, SC-19192:
115 Week Oral Tumorigenicity Study in the Rat, are as follows:

A.  Design and Conduct of Study

1)  Control and treated animals were randomly distributed
    on the same rack. (See diagram of housing group
    attached as exhibit 7.)

2)  No ear clips or other methods of uniquely identifying
    each animal were used.  Identification consisted of two
    types of cards attached to the front of each cage.

3)  Compound inventory cards were deficient in that only one of
    18 such cards stated the purpose (study 988S73) for with-
    drawing the compound from inventory.  Three of the cards
    did not include the date withdrawn, amount withdrawn, or
    signature of requestor.  Therefore it was impossible to
    Reconcile the amount withdrawn and the amount used.
    (See exhibit #28.)

4)  Food jars were not individually identified, yet all the
    filled jars for a given housing group (control, low, mid,
    and high dose) were placed on a mobile cart, which was
    wheeled to the housing rack.  The position of the jar
    (in rows) on the cart was the only means of identifying
    the proper dose level.  The arrangement of the food cups
    on the cart is shown in exhibit #8.

5)  A total of 79 "observations for drug effects" records were
    not signed or initialed.

6)  Observation records indicated that animal A23LM was alive at
    week 88, dead from week 92 through week 104, alive at week
    108, and dead at week 112.

7)  Records indicated that at the scheduled 104 week bleeding,
    animal E2CM was substituted for animal A11CM.  Records also indica-
    ted that animal A11CM was alive on this date and therefore
    should have been bled as scheduled.

8)  Records indicated that penicillin was administered to four
    rats beginning on May 16, 1973, and continuing daily through
    May 28, 1973.  This third occurrence of infections disease
    and penicillin administration was not reported in the sub-
    mission to FDA.

                           (2)

9)  In many cases the actual number of tissues embedded was less
    than the 24 (control and high dose) or 19 (low and mid dose)
    specified in the final histology lab protocol dated 1/21/74.

10) Ophthalmoscopic examination records were present for animals
    H26MF and J29CM, yet the findings were not reported in the
    submission to FDA.  Two other discrepancies of this type
    were noted.

11) Records indicate that a tissue mass measuring 1.5 x 1.0 cm
    was excised from animal B3HF on 2/12/72, and that a "skin
    incision over mass" was performed on animals C22LM and G25LM
    on Feb. 10, 1972.

B.  Stability and Homogeneity of DKP in Diet Mixtures

1)  There were no batch records to show the quantities of DKP
    and basal diet weighted, type of mixer used, mixing time,
    dates, or names of individuals performing the weighing and
    blending operations.

2)  There was no evidence that any tests had been done to deter-
    mine the blending characteristics of the mixer, or to vali-
    date the mixing time.

3)  No homogeneity tests were performed on any batches of diet
    used in the study, and to stability study assay reports
    (A7738 and A7739) indicated that samples were not homogene-
    ous.  (See exhibit #29.)

4)  A stability study was conducted with DKP
    in 1972.  However, the 115 week rat stud employed
    Basal Diet from week 2 to its conclusion, and to stability
    studies had been conducted with Basal Diet.

5)  Methods of assay for DKP in the diet were deficient in that:
    The titration method was discontinued after 1 week of the
    stability study.  Some of the TLC photographs showed no DKP
    reference standards and photographs also showed that there
    was something in the basal diet itself producing a spot
    on the TLC plate which had an Rf value corresponding to
    DKP.  Only one solvent system was  used for development of
    the TLC Plates.  Some of the chromatograms showed poor separa-
    tion.

                           (3)

6)  No reserve samples of any of the lots of DKP used in this study
    were retained by Searle.

7)  Three different sets of specifications for DKP were found, and
    Searle could not determine with any degree of certainty which
    of the three were applicable to the 7 lots of DKP used in the
    study.

8)  The analytical records for DKP lots IR through 5R refer to
    reference standard IR #3701.  None of the three sets of DKP
    specifications lists reference #3701.  No data was made avail-
    able as to dates, methods of preparation and authentication
    of DKP reference standards.

9)  Analytical records A-9129 for DKP lot 5R showed an assay of
    1000%.  Examination of laboratory notebooks showed
    that eleven (11) samples had been analyzed from this lot, and
    the analytical record only reflected an average of the last
    three of theses.  The other assays (not reported) ranged from
    87.93% to 114.83%.

C.  Dosage, Body Weight and Food Consumption

1)  Examination of the raw data sheets revealed the following
    discrepancies:

   a.  Empty feed cup weights were missing for the D hous-
       ing group at the 12th week, in the raw data sheets.  (See
       exhibit #75.)

    b. In several instances, the dietary concentration shown
       on the weight sheets did not agree with the concentra-
       tion listed for the same level in the other housing
       groups.  (For example; C group Males, mid & high levels
       for week 13,; A group Males, high levels for week 99.)

2)  Comparison of the Searle submission and the independent FDA
    analysis of the raw body weight and food consumption data
    revealed the following discrepancies:

    a. We found a total of 15 differences of 1 gram or more in
       the average body weight and of 0.1 percentage points
       or more in weight gain.  (See table 1.)

                           (4)

    b. We found approximately 82 discrepancies of one gram or
       or more in the food intake when expressed in grams/day.
       (See  table 2.)

    c. We found approximately 40 errors of 5 or more grams in
       food intake when expressed in grams/kg./day.  (See
       table 2.)

    d. Most of our dosage calculations differed from Searle's
       dosage calculations by 10 or more mg., when the dosage
       is expressed as mg/kg/day.  (See table 2.)

D.   Gross and Microscopic Pathology

1)  98 of the 196 animals that died during the study were fixed
    in toto and autopsied at some later date, in some cases
    more than one year later.

2)  A total of 20 animals were excluded from the study due to
    excessive autolysis.  Of these, 17 had been fixed in toto
    and autopsied at a later date.

3)  Records indicated that animal F6HF, a high dose female, was
    found dead at 787 days of treatment and the gross pathology
    sheet reported a tissue mass measuring 5.0 X 4.5 X 2.5 cm.
    The submission to FDA reported no tissue mass and the
    animal was excluded from the study due to marked autolysis.

4)  Records for approximately 30 animals showed substantial
    differences between gross observations on pathology sheets,
    when compared with the gross observations on pathology sheets
    submitted to FDA.  A detailed description of 10 of these is
    included in the report.  Copies of all the gross pathology
    sheets, and the pathology summaries submitted to FDA are
    attached as exhibits.

5)  Dr. Charles H. Frith, D.V.M., Ph.D., Directory, Pathology
    Services, NCTR, examined slides for a total of 150 animals,
    or about 42 percent of the animals on study.  He noted
    the following discrepancies:

    a. The reporting of a mass (by Searle) as missing which was
       actually present (animal M1LF.)

                           (5)

    b. The finding of a polyp of the uterus which was not
       diagnosed by Searle (animal K9MF).  The finding of
       this additional uterine polyp by Dr. Frith increases
       the incidence in the midi dose to 5 of 34. (15 percent.)

    c. The finding of ovarian neoplasms in animals H19CF, H19C,
       and H7HF, and the finding of diffuse hyperplasia in animal
       D29CF, which were not diagnosed by Searle.

    d. The finding of additional inconsistencies in 21 animals.

6)  No microscopic worksheets or other "raw data" relating to
    microscopic pathology could be found for this study.

7)  A mammary tumor found in animal F27CF was described as a
    papillary cystadenoma on the pathology summary sheet, (page
    105, Vol. II of the submission) and as an adenocarcinoma on
    summary table 12 (p. 95, Vol. I of the submission).

8)  In several instances the histopathology technician made notes
    at the bottom of the gross pathology sheet to indicate that
    certain organs were not present in the bottle of fixative
    (and were therefore not available for sectioning).  Yet, in
    three of these instances (animals A4CM, K23CF, and J3CM) a
    diagnosis appears in the submission to FDA.

E.   Organ Weights

1)  Organ weights were entered on the gross pathology sheets at
    the time of autopsy.  We compared all of the individual organ
    weights on appendix table 5 in the submission to FDA (Vol. 1,
    pgs. 222-226) with the original data on the gross pathology
    sheets.  A total of eleven (11) errors were noted in transcrib-
    ing the raw data from the pathology sheets to the tables in the
    submission to FDA.

F.  Survival

1.  We were  unable to determine the exact method used by Searle in
    constructing the survival table in the submission to FDA.
    We constructed a survival table using the body/feeder weight
    Teletype sheets.  A Life Table Analysis was constructed from
    our survival table by Dennis Wilson, FDA Department of Mathe-
    matics.  The female control population differed from the high
    level population (p 0.05) and the mail control population
    differed from the mid and high level population (p 0.05).  In
    all cases the differences are due to higher mortality in
    the controls.

                           (6)

G.  Clinical Laboratory Procedures

1. Laboratory records of one sort or another for all assays report-
   ted in the submission were obtained.  In some cases data sheets
   were noted with results of assays carried out at treatment days
   not indicated in the protocol or protocol amendment.  For example,
   serum cholesterol determinations were done at days 796 and 798
   (terminal bleeding) but not included in the submission to FDA.
   Because the submission to FDA (Vol. 1 p. 286) reported a signi-
   ficant decrease in serum cholesterol that was more perceptible
   towards the end of the study, and may have been related to
   compound administration, the omitted data is of some importance.

2.  No data was seen for two assays (serum insulin and serum ornit-
   hine carbamyl transferase) which were called for in an amend-
   mend to the protocol.

3.  Original data was not always available for authentication of
   results or examination of procedures for conversion of raw
   data into the calculated values submitted to FDA.

4.  Data pages for clinical chemistry and urinalysis were initialled
   by a technician who transcribed data but apparently was not
   directly involved in the assays indicated.  He stated in an
   interview that Dr. Rao told him to initial the data sheets.

5.  The methodology as referenced in the submission to FDA is incom-
   plete and not always an accurate reflection of the methodology
   actually used in the study.  The fact that more than one method
   was sometimes used for a particular assay during different times
   of the study was not indicated in the submission to FDA.

6.  A total of 21 disparities between individual clinical laboratory
   analysis values appearing in the submission Volume I and those
   values appearing in data sheets and/or laboratory notebooks were
   found.

7.  A  total of 49 disparities were noted  between statistical
   computations reported by Searle in the submission and
   those calculated by FDA.  The disparities are constituted
   by the values for 6 means, 23 standard errors, and 20 signi-
   ficant differences (as measured by T tests).

8.  Some of the data sheets for urinalysis had erroneously
   labeled the phenylketones test values as "phenylalanine".

                           (7)

PURPOSE OF INVESTIGATION

Assignment memo dated may 16, 1977 from Donald Healton, Acting
Director of Regional Operations, confirmed an earlier oral
assignment to Chicago District for a directed inspection of cer-
tain non-clinical studies submitted to FDA in  support of a food
additive petition for the sweetener aspartame.

The investigation began on 4/25/77, and encompassed the authen-
tication of all data, both raw and summary, relating to the
studies jointly chosen for review by Bureau of Foods and EDRO.
Two studies actually done at G.D. Searle were selected for initial
coverage, and a decision to expand the investigation to a third
study was made at a later date.

Following are the titles of the three studies selected for review:

1.)  E-5 (P.T. #851S70), Evaluation of the Embryotoxic and Teatogenic
    Potential in the Rat, conducted with SC-18862 (aspar-
    tame).

2.)  E-89 (PT #1218S75), An evaluation of the Embryotosic and Tetato-
    genic Potential in the Mouse, conducted with SC-18862 (aspar-
    tame).

3.)  E-77/78 (PT #988S73), 115 Week Oral Tumorigenicity Study in the
     Rat, conducted with SC-19192 (diketopiperazine).

This report is concerned only with study E-77/78.  The report of
E-5 and E-89 was submitted separately.

HISTORY OF BUSINESS

G. D. Searle & Co. provides a wide range of health care products
and services on a worldwide basis.  Its business is divided
among three principal areas:  pharmaceuticals, medical instru-
ments and optical products, and hospital and laboratory products.
The firm's corporate offices are located in Skokie, Illinois,
with various branches and facilities throughout the world.

Effective June 1, 1977, Donald H. Rumsfeld assumed duties as
President and Chief Executive Officer.  Mr. Daniel C. Searle,
formerly Chief Executive Officer is now Chairman of the Board,
while William L. Searle and Wesley M. Dixon, former Chairman
and President respectively, are now Vice-Chairmen.

                           (8)

Effective March 1, 1977, the firm underwent a major realignment,
shifting to a managerial system based on product lines.  This
resulted in the establishment of four main product-line groups,
which are:  Pharmaceutical/Consumer Products, Diagnostics,
Hospital Supplies and Optical Products.  Each group is headed
by a President who will report to Searle's Executive Vice-Presi-
dent for Operations, Dr. James A. Buzard.  A copy of the G. D.
Searle & Co. annual report for 1976 which is attached as Exhibit
#1 further expands on the firm's operations and lists Corpor-
ate Officers.

Mr. O. B. Parrish is President of the Pharmaceutical/Consumer
Products Group and also a Corporate Vice-President.  An organi-
zational chart for this group is attached as Exhibit #2.  Mr.
Guy Labrosse is now Group Executive Vice-President for U. S.
Commercial Pharmaceutical Operations.  In the U. S., this is known
as Searle Laboratories.  The facility at 4901 Searle Parkway,
Skokie, Illinois is a part of the U. S. Operations, e. g. Searle
Laboratories, yet houses the majority of the Research and
Development Group.

Worldwide Pharmaceutical Research and Development is also a
part of the Pharmaceutical/Consumer Products Group, but not of
Searle Laboratories.  The Research and Development OF aspartame
is a function of this group.  Copies of organizational charts
for this group are attached as Exhibit #3.  Dr. Robert A. Moe
recently resigned and his position is temporarily being filled
by George V. O'Bleness, Corporate Vice-President for Compliance
and Administration.

Commercial aspects of Aspartame are being handled by an "aspar-
tame Division", under the direction of Elwood H. Ensor, Corpor-
ate Vice-President.  There is no longer a division entitled
"New Ventures".

PERSONS INTERVIEWED

Credentials were shown and a written Notice of Inspection was
issued to Dr. William M. Merino, Directory, Domestic Pharmaceu-
tical Products, Regulatory Affairs Department on April 25,
1977.  The following Searle personnel were present at the
initial meeting on 4-25-1977.

                           (9)

Robert A. Moe, PhD. - Executive Vice-President
George Clay, PhD. - Group Leader, CNS Pharmacology
Robert Bost, PhD. - Director of Food Products,
                   Regulatory Affairs
Holly Ru Probst - Director, Corporation Information
                  Management Group.
Dave Britton - Director Corporation Information
               Department
William Merino, PhD. - Director, Domestic Pharmaceutical
                Products
Richard Viktora - Attorney
James Phelps - Vice-President, General Counsel
Elwood H. Ensor, PhD. - Vice-President
Paul Klimstra, PhD. - Vice-President Pre-Clinical
                      Research and Development
Roger Thies - Attorney

During the course of our investigation one or more of the
following Searle personnel were present in the Conference Room
which we used for our data review.

  Richard Viktora - Attorney
  Roger Thies - Attorney
  George Clay, PhD. - Group Leader, CNS Pharmacology
  Robert Bost, PhD. - Director of Food Products
                      Regulatory affairs
  Don Cook, PhD. - Associate Director, Department of
                   Bio Research
  Dick Aspinol, PhD. - I. I. D. Group Leader
  Bill Jenkins, PhD. - Director, Product Affairs
  Fred McIlreath, PhD. - Director, Regulatory Affairs
  Paul Landefeld, - Attorney

Most of the time one attorney (R. Viktora or R. Thies) and
one scientist were present.  During our initial meeting with
Searle personnel, James Phelps stated that a Searle monitor
must be with us at all times during our data review in order
to "protect the data".

During the course of our investigation, various individuals
were interviewed in an attempt to obtain all available raw
data and reconstruct the manner in which the study was con-
ducted, as accurately as possible.  Since many employees
involved in the study or support areas are no longer employed
at Searle, others were interviewed who had general knowledge
of such parameters as statistics and chemistry.



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