XENOBIOTICA. 1973, VOL. 3, NO.

11, 711-714

The Metabolism of Tamoxifen” (I.C.I. 45,474)

Part 11 : In Female Patients
JOHN M. FROMSON, STUART PEARSON and
SARA BRAMAH (nee MEEK)
Imperial Chemical Industries Ltd., Pharmaceuticals Division, Alderley Park,
P.O. Box 25, Macclesfield, Cheshire, U.K.

(Received 2 April 1973)

1. The metabolism of orally administered [“Cltamoxifen was investigated in

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four women. Most of the 14C was slowly excreted in the faeces, only small
amounts appearing in the urine.
2. The labelled material in the faeces was present mainly as conjugates.
Unchanged drug and hydroxylated metabolites accounted for less than 30% of
the faecal radioactivity.
3. Blood levels of total radioactivity following oral doses of about 0.3 mg/kg
reached peak values of 0.06-0.14 pg/ml 4-7 h after dosing. At this time only
20-30% of the radioactive material was present as tamoxifen.
4. The prolongation of blood levels and faecal excretion is thought to be due
to enterohepatic circulation.
For personal use only.

Introduction
The metabolism of tamoxifen in laboratory animals has been discussed
(Fromson, Pearson & Bramah, 1973). For human studies, women with mam-
mary carcinoma and women already receiving the compound for the treatment of
dysfunctional uterine bleeding were, with their informed consents, each given
a single oral dose of 14C-labelled tamoxifen. I n some of the latter patients the
proportion of the labelled dose which became bound to uterine tissue was investi-
gated following hysterectomy (Fromson & Sharp, unpublished observations).
The present studies were directed towards obtaining data concerning the absorp-
tion, metabolism and excretion of tamoxifen in these patients.

Materials and methods

[14C]Tamoxifen of specific activity 24.6 pCi/mg was used throughout the
study, The preparation of biological samples for chromatographic and radio-
activity counting was similar to that described in the preceding paper (Fromson
et al., 1973) except where otherwise stated.
Dosage of drug, collection and treatment of samples
All patients received doses of the l’C-labelled drug in gelatine capsules (20 mg
tamoxifen containing 40 pCi [l‘C]tamoxifen). I n the patients who underwent
hysterectomy the capsule was dosed approximately 5 h before operation.
Complete 24 h urine and faeces specimens were collected from one subject
for 8 days and from another for 13 days, following administration of the radio-
active drug.
‘ NOLVADEX ’, Trade Mark, the property of Imperial Chemical Industries Limited.
3E2
 712 J. M. Fromson et al.
Isolation and identification of metabolites
Urine. It was not possible to obtain significant results from the t.1.c.
examination of urine due to the low amounts of 14C-labelled material excreted by
this route.
Faeces. Individual faecal extracts were examined by t.1.c. in solvent systems
A and B (see previous paper) and the separated radioactive components quanti-
tated by scintillation counting of the segments. The 14C-material in the com-
bined extracts was partially separatedfrom non-radioactive material by percolating
through an Amberlite XAD-2 column. T h e material was then purified by
preparative t.1.c. and the resultant fractions investigated by g.1.c.-mass spectro-
metry.
Serum. In an attempt to assess the relative proportions of unchanged drug
and metabolites in the serum, samples from Patient 2 were combined as indicated
in Table 1. Methanol was added and the precipitated protein extracted ex-
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haustively with more methanol until all the radioactivity was removed. After
centrifugation the extracts were combined, evaporated and investigated by t.1.c.
in solvent system A.

Table 1. Percentages* of lT-labelled components in human serum following a

single oral dose of [14C]tamoxifen
Serum samples combined % % %
(hours) Unchanged drug Metabolite B Origin material
For personal use only.

1 and 4 33 9 30
6, 8 and 10 8 15 42
13, 19, and 25 11 21 28
37, 53 and 75 4 20 33
99,123 and 172 4 18 47
* Results from t.1.c. plates developed in solvent system A and expressed as a percentage
of the total radioactivity present.

Results
Excretion of 14C-labelled material
Owing to very slow excretion of the compound, non-quantitative recoveries
were obtained from both patients. Over an 8 day sampling period one patient
excreted 9% of the dose in the urine and 26% in the faeces. Sampling for 13 days
from a second patient resulted in recoveries of 14% in the urine and 51% in the
faeces. I n the latter patient, post-operative constipation prevailed for the first
5 days. From the data available it was possible to calculate that material was
eliminated between the fifth and eighth day with a half-life of about 4 days,
increasing to about 9 days by day 13.
Serum levels and half-lives
T h e blood level profile from Patient 2 is illustrated in the figure. Levels are
expressed as pg tamoxifen/ml serum assuming the material present to have a
molecular weight similar to that of tamoxifen. The maximum serum levels were
achieved 4 h after dosing. Levels then decreased with an initial half-life of 11 h.
There were indications of smaller peaks at 4 days and 10 days after dosing which
may be attributable to enterohepatic circulation. At 14 days a serum level
equivalent to 0.013 f 0.002pg tamoxifen was observed.
 Metabolism of Tamoxifen by Humans 713
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0 1 2 3 4 5 6 7 8 9 1011 1213 14
Time (days)
Levels of total radioactivity in serum after oral administration of [14C]tam~xifen
to a female
patient.
Dose was 0.37 mg/kg.

I n three other patients dosed at the same level ( 0.3 mg/kg) maximum serum
N
For personal use only.

concentrations of 0.06-0.1 pg tamoxifen/ml were reached 4-7 h after administra-

tion. After initial half-lives of 7-14 h serum levels decayed with secondary half-
lives greater than 7 days.
Examination of serum
The percentages of unchanged drug, metabolite B and origin material in the
methanolic extracts of the combined groups of serum samples from Patient 2 are
shown in Table 1.
Examination of faecal extracts
The percentages of material with R, values similar to tamoxifen, metabolite
B, metabolite F (see preceding paper for structures) and origin material (rep-
resenting polar conjugates) are given in Table 2. None of the other metabolites
found in the animal experiments were observed.

Discussion
The prolonged blood levels and long excretion half-life of total radioactivity
found in laboratory animals (Fromson, et al., 1973) are also observed in the pre-
sent human studies. Two weeks after receiving a labelled dose only 65% of the
dose could be accounted for, although radioactivity was still detectable in the
blood.
An analogous situation was encountered when [14C]clomiphene citrate was
administered orally to human subjects (Schreiber, Johnson, Plotz & Wiener,
1967). Over 5 days only 51% of the dose was excreted and radioactivity was
found in the faeces up to 6 weeks after administration.
Most of the labelled material in the faeces was present as polar conjugates.
The amount of unchanged drug in the faeces appeared to decrease over the period
 714 -7.M . Frontson et al.
Table 2. Percentages* of l4C-1abelledcomponents in extracts of human faeces
following a single oral dose of [14C]tamoxifen

Time of faecal % % % %
sample (days)? Unchanged drug Metabolite B Origin material Metabolite F
~

6 (morning) 16 8 52 7
(evening) 11 6 60 . 5
7 20 3 52 7
8 12 4 54 8
11 4 2 67 7
13 4 3 61 6

* Results from t.1.c. plates developed in solvent systems A and B and expressed as a
percentage of the total radioactivity present.
t Faecal samples from Patient 2.
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of study as did that of the monohydroxylated metabolite (metabolite B), whereas

the percentages of conjugated material and metabolite F remained about the same.
The presence of metabolite F was perhaps surprising as previously it had only
been found in rat bile. None of the other metabolites isolated in the animal
studies was detected.
T h e speed with which the compound is metabolized is reflected in the con-
centrations of tamoxifen in the serum relative to the amounts of total radioactivity.
I n the first few hours 330/, of the radioactivity was attributable to the parent
compound but levels decreased until only 4% of the radioactivity was present
For personal use only.

as unchanged drug 7 days after dosing. Variable amounts (28-47%) of origin

material were obtained over this period.
In animals enterohepatic circulation of radioactivity was evident and it was
postulated that the circulating material largely consisted of hydroxylated meta-
bolites (Fromson, et al., 1973). It is probable that a similar situation exists in
humans, as suggested by the prolongation of blood levels and the presence of some
of the hydroxylated metabolites in the faeces. Some of these compounds have
been shown to possess anti-oestrogenic activity. The corresponding glucuronides
may represent a storage form of anti-oestrogenic activity in the gastrointestinal
tract from which the hydroxylated metabolites can be liberated enzymically.
Thus, biliary recirculation could prolong the duration of anti-oestrogenic activity
and may contribute to the clinical efficacy of the compound in mammary
carcinoma.

Acknowledgments
T h e authors wish to acknowledge Dr. I. D. H. Todd (Christie Hospital,
Manchester), Dr. A. Klopper (Foresterhill Hospital, Aberdeen) and Drs. R. W.
Burslem and D. S. Sharp (Withington Hospital, Manchester) for their co-opera-
tion in the study.
We are also indebted to the following I.C.I. personnel : Mr. J. Burns and
Mr. D. Potts for the preparation of [14C]tamoxifen,Mr. J. Webster for g.1.c.-mass
spectral determinations and Mr. T. Helliwell for technical assistance.

References
J. M., PEARSON,
FROMSON, S. and BRAMAH,
S. E. (1973). Xenobiotica, 3,693.
E., JOHNSON, J. E., PLOTZ,
SCHREIBER, E, J. and WIENER,
M. (1967). Clin. Res., 14, 287.