Br. J. Cancer (1983), 47, 035-042
`
`Localisation and toxicity study of a vindesine-anti-CEA
`conjugate in patients with advanced cancer
`Ford'*,
`C.E. Newman'*, J.R. Johnson1,
`C.S. Woodhouse',
`C.H.J.
`Reeder2t, G.F. Rowland3 & R.G. Simmonds3
`'Surgical Immunology Unit, Clinical Oncology, 2the Dept. of Nuclear Medicine, Queen Elizabeth Hospital,
`Birmingham, and 3the Lilly Research Centre, Erl Wood Manor, Windlesham, Surrey.
`
`T.A.
`
`Summary Safety of administration of a vindesine (VDS)-anti-CEA conjugate and its ability to localise after
`radiolabelling were investigated in patients with advanced metastatic carcinoma (4 colorectal and 4 ovarian).
`For imaging, patients received between 230 and 520pg of 13 1I labelled antibody. In 5, localisation of
`conjugate was demonstrated, in another it was equivocal and in 2 patients, undetectable. For assessment of
`safety each patient also received a single dose of conjugate increasing from 1.2 to 42mg antibody linked to 24
`to 1800 g VDS. The in vitro activity of the anti-CEA antibody and its ability to localise in vivo were
`preserved after conjugation. There was no obvious toxicity or hypersensitivity attributable to either the
`radiolocalisation or escalated doses of conjugate in any of the patients. The feasibility of the preparation and
`administration to patients of a vindesine-antibody conjugate has been demonstrated.
`
`The concept of targeting drugs on to malignant
`cells proposed by Ehrlich (1900) offers a potential
`improvement over one of the major limitations of
`cancer chemotherapy, viz. the limited selectivity of
`drugs for cancer cells. The use of antibodies as
`carriers was tested experimentally in an L1210
`leukaemia (Mathe et al., 1958) and clinically in
`malignant melanoma (Ghose et al., 1972).
`Despite
`of some success
`reports
`with
`this
`approach it has not been widely adopted. This has
`been partly due to the inability to demonstrate
`tumour-specific
`antigens
`in human
`targets
`as
`however,
`tumours;
`in
`order
`increase
`the
`to
`therapeutic index of a drug, differential expression
`of the target by the tumour compared to normal
`tissue may be sufficient. The well characterised
`tumour-associated
`antigens,
`carcinoembryonic
`antigen (CEA) and alpha fetoprotein (AFP), offer
`such potential targets in man. Another reason for
`caution with this approach has been scepticism
`about the in vivo stability of the drug and antibody
`conjugate. It was demonstrated in the mouse EL4
`lymphoma model, that drug and antibody were
`interactive and more effective than a conjugate of
`the two (Davies & O'Neill, 1973). This led to a
`pilot study of drug and antibody interaction in
`patients
`with
`bronchogenic
`resected
`carcinoma
`
`*Present
`address:
`Oncology
`Research,
`Memorial
`University and Newfoundland Cancer Clinic,
`Health
`Sciences Centre, St. John's, Newfoundland, Canada AlB
`3V6.
`$Present address: Medical Physics Department, North
`Tees General Hospital, Stockton-on-Tees, Cleveland.
`
`Received 29 June 1982; Accepted 23 September 1982.
`
`0007-0920/83/010035-08 $01.00
`
`(Newman et al., 1977) and to studies of a variety of
`drug and antibody conjugates and intermediate
`carriers (Ghose & Blair, 1978; Lee & Hwang, 1979;
`Dullens & De Weger, 1980, and Rowland, 1982).
`The demonstration
`of enhanced
`toxicity
`of
`vincristine for a CEA-secreting lung cancer cell line
`in the presence of anti-CEA-immunoglobulin (Ig)
`(Johnson et al., 1980) encouraged us to investigate
`direct conjugation of vinca alkaloids to antibody.
`The toxicity of vindesine for a CEA-secreting cell
`line was found to be greatly
`increased when
`conjugated to an anti-CEA-Ig (Johnson et
`al.,
`1981). The aims of the present study were to
`investigate the ability to localise and safety of
`administration of this conjugate in patients with
`advanced metastatic adenocarcinomas refractory to
`established forms of treatment.
`
`Materials and methods
`Patients
`Eight patients with advanced metastatic carcinoma
`refractory to previous treatment were entered into
`study;
`this
`all gave informed consent.
`Patients
`selected had tumour types likely to express CEA
`and to localise anti-CEA-antibodies (Goldenberg et
`al., 1978a; Dykes et al., 1980; Van Nagell et al.,
`1980). Four had disseminated ovarian carcinomas
`and 4 had disseminated colorectal carcinomas.
`Before injection of radio-labelled antibody, patients
`tested
`for
`immediate
`delayed
`and
`were
`hypersensitivity to sheep Ig (0.1 ml of mg ml- 1).
`When there was a delay of more than a week
`between the first dose of conjugate and the next
`dose sensitivity
`testing was repeated. To block
`
`The Macmillan Press Ltd., 1983
`
`IMMUNOGEN 2297, pg. 1
`Phigenix v. Immunogen
`IPR2014-00676
`
`

`

`36
`
`C.H.J. FORD et al.
`
`thyroid uptake of 131 I, potassium iodide tablets
`(180mgday-1) were given, beginning
`1-3 days
`before
`administration
`of
`the
`iodinated
`conjugate/antibody and continuing for 8-12 days
`thereafter.
`Patients were admitted to hospital 1-3 days prior
`to the study for complete physical examination,
`laboratory
`investigations
`baseline
`and
`hypersensitivity testing. Venous blood was sampled
`full
`for
`blood
`electrolytes,
`and
`count,
`urea
`biochemical profile and GTs. Twenty-four hour
`creatinine
`clearances
`performed.
`also
`were
`pulse
`blood
`Temperature,
`and
`pressure
`were
`half-hourly
`recorded
`during
`and
`for
`4-6 h
`immediately after infusion of conjugate and every
`4 h thereafter. Subjective toxicity was monitored
`daily by the attending physician (JRJ) and routine
`follow-up investigations performed every 1-2 days
`for the first 10-14 days. Particular attention was
`paid to evidence of hypersensitivity reactions and
`neurological status on clinical examination. Patients
`were then followed up in the clinic
`at weekly
`intervals for a minimum of 1 month, and in most
`cases 2-3 months.
`
`Antibody
`Immunoglobulin (Ig) was prepared from sheep anti-
`CEA serum by ammonium sulphate precipitation
`and
`by
`provided
`A.R.
`Bradwell,
`Dr.
`was
`Immunodiagnostics
`Laboratory,
`Research
`University of Birmingham, U.K. The antibody had
`been absorbed with normal liver, colon, lung and
`spleen. In fused rocket immunoelectrophoresis it did
`not recognise the CEA cross-reacting determinants
`shared with the non-specific cross-reacting antigen
`(NCA). Localisation of this antibody to human
`gastrointestinal tumour deposits has been reported
`(Dykes et al., 1980). In our hands it localised on
`sections of a formalin-fixed CEA-secreting colonic
`carcinoma in an indirect immunoperoxidase test at
`of
`titre
`1/80,000.
`a
`However,
`in
`the
`immunocytochemical tests there was still residual
`antibody activity to shared NCA determinants as
`demonstrated
`by staining
`of
`chronic
`myeloid
`leukaemia cells and splenic myeloid cells (Ford et
`al., 1981).
`
`Ig
`
`Conjugate
`Vindesine
`(VDS)-anti-CEA
`conjugates
`were
`Lilly
`prepared
`Research
`at
`from
`Centre
`Ltd.
`desacetylvincaleucoblastine
`acid hydrazide under
`aseptic
`by
`conditions
`modification
`the
`of
`a
`procedure described for vindesine-BSA (Conrad et
`al., 1979) and purified by gel filtration. Four batches
`were prepared with initial conjugation ratios of 4.1,
`5.4, 4.3 and 11 moles vindesine per mole IgG. An
`
`iodinated aliquot of Batch I was used to scan
`patients 1 and 2. Iodinated Batch II was used to
`scan patients 3-6. Patients 7 and 8 were scanned
`with an iodinated aliquot of the sheep anti-CEA Ig
`used to prepare Batch IV.
`For assessment of safety, patients received doses
`of 1.2-42mg conjugate, containing 24-1800pg VDS,
`injected i.v. in 100ml of 1% human serum albumin
`in saline (HSA-saline) over a 30-60min period. All
`conjugates were 0.22pm filtered before dilution in
`sterile, HSA-saline.
`Radiolabelling of conjugate/antibody
`Aliquots of batches of conjugate (or antibody) were
`iodinated with 1311 using a modified chloramine-T
`method (Garvey et
`al.,
`1977). Free iodine was
`removed on a Sephadex G-25 column which was
`eluted with HSA-saline. Fractions containing the
`protein peak as determined by y-counting were
`pooled and sterile filtered through a 0.22,pm filter.
`Radiolabelled conjugate (or antibody) was injected
`i.v. in 100ml of sterile HSA-saline over a 30-60min
`period.
`All solutions were tested for pyrogenicity and
`sterility.
`lodinated
`conjugates
`tested
`were
`at
`6pgIgkg-1 body weight in rabbits and uniodinated
`conjugates
`tested
`from
`were
`45-198 pg Ig kg-1
`body weight
`depending on
`the
`dose
`to
`be
`administered to patients. All batches were negative.
`Photoscanning
`Between 230-520pgIg conjugate (6-14 pg of VDS)
`containing 541-1014pCi protein bound 131I was
`administered to patients 1-6. Patients 7 and 8 each
`471 pg of unconjugated Ig
`received
`containing
`1056 pCi 1311.
`Each patient
`received
`i.v.
`99mTc-pertechnetate
`(500 pCi) 30 min before each scan, and 99mTc-
`labelled human serum albumin (500 pCi)
`5 min
`before each scan. These distribute similarly to free
`iodide and radiolabelled antibody respectively in
`the blood pool. Images of the chest and abdomen
`were obtained with a gamma camera (Searle LFOV
`with medium energy collimator) initially at 4, 24
`and 48 h after injection of iodinated material. The
`4 h scans were discontinued for patients 3-8. The
`camera was linked to a DEC PDP1 1/40 computer
`with a dual isotope facility and a colour scale visual
`display unit. The data were stored and displayed in
`a 64 x 64 matrix. After normalising over the cardiac
`area, subtraction of the technetium component from
`the iodine component was performed to visualise
`areas of selective uptake of conjugate.
`
`Measurement of anti-CEA activity
`A modified enzyme-linked immunosorbent assay
`
`IMMUNOGEN 2297, pg. 2
`Phigenix v. Immunogen
`IPR2014-00676
`
`

`

`A VINDESINE ANTI-CEA CONJUGATE IN VIVO
`
`37
`
`(ELISA) (Woodhouse et
`1982) was used to
`al.,
`measure anti-CEA activity of antibody, conjugate
`and iodinated conjugate. Briefly, disposable cuvettes
`were coated with purified CEA, washed, blocked by
`incubation with BSA, washed and the test sample
`added, before incubation at 35°C for 3 h. Following
`further
`anti-sheep
`washing,
`rabbit
`a
`IgG
`horseradish peroxidase conjugate (Nordic, U.K.)
`was added. The cuvettes were incubated at 35°C for
`washed
`3 h,
`then ABTS (2.2-azino-di-(3-
`and
`ethylbenzthiazoline sulphonic acid) (Sigma)) was
`added. Absorbance at 405 nm was measured using a
`Gilford PR-50 processor-reader.
`
`Measurement of serum CEA levels
`CEA measurements were performed by Dr.
`P.
`Gosling,
`Birmingham
`Hospital,
`using
`East
`a
`modified double antibody technique (Booth et al.,
`1973). Serum samples were perchloric acid-extracted
`before assay.
`
`Results
`
`The dosages of radiolabelled conjugate/antibody
`and uniodinated conjugate received by each patient
`are given in Table I.
`Details of the patients in this study and the
`scanning
`are summarised in Table
`results
`II.
`Patients
`radiolabelled
`received
`conjugate,
`1-6
`followed within 4-54 days by unlabelled conjugate.
`Localisation of radioactivity which equated with
`clinically detectable disease was seen in patients 1,
`2, 3, 4 & 6. The localisation picture for patient 5
`was equivocal. Figures 1 and 2 illustrate localisation
`images. Figure 1
`is the 48h subtraction scan for
`patient 2 who had a large abdomino-pelvic mass
`
`which was confirmed on CT scans. Localisation of
`isotope occurred in the mass and in the right
`kidney. Subsequent investigation of this patient by
`intravenous
`pyelography
`indicated
`right
`a
`hydronephrosis due to compression of the right
`by
`the
`Impaired
`excretion
`ureter
`tumour.
`apparently resulted in an accumulation of isotope in
`the right kidney.
`Figure 2 is the 48 h subtraction scan for patient 6
`who had an extensive pelvic tumour mass and
`palpable abdominal masses.
`central
`Ultrasound
`examination confirmed the presence of enlarged
`para-aortic and coeliac axis nodes. Localisation of
`isotope coinciding with these, and the pelvic mass
`at the primary site, can be seen in Figure 2.
`For practical reasons, the last 2 patients received
`uniodinated
`conjugate
`and
`then
`radiolabelled
`unconjugated antibody. There was no evidence of
`localisation in these patients (see Discussion).
`The relative anti-CEA activities
`of the non-
`iodinated conjugates in ELISA compared to the
`original antibody, when tested within 14 days of
`conjugation, were: Batch I, 98%; Batch II, 100%;
`Batch III, 80%. In the case of Batch IV it was not
`possible to obtain a relative anti-CEA value. After
`iodination the values were: Batch I, 48%; Batch II,
`72%. Batches III and IV were not iodinated and the
`radiolabelled antibody used for patients 7 and 8
`had 70% activity.
`With the exception of patient 6, all had raised
`(>15 ngml- 1)
`pretreatment
`serum CEA levels
`(Table 2). We noted no significant decline in CEA
`levels in any of the patients after the radiolocalising
`dose. One day after administration of 11.06mg
`VDS-Ig conjugate to patient 3, there was a fall from
`31-17ngml-' which was sustained for
`days.
`3
`Similarly, for patient 6, who developed a raised
`CEA level of 37ngml-1 from a pre-treatment value
`
`Table I Summary of dosages
`
`VDS
`ug
`
`6
`6
`6.25
`6.5
`14.1
`14.1
`
`Patient
`
`1
`2
`3
`4
`5
`6
`7
`8
`
`Iodinated antibody
`Ig
`9g
`300
`300
`230
`240
`520
`520
`471
`471
`
`1311*
`pCi
`
`996
`541
`1014
`581
`633
`633
`1056
`1056
`
`Non-iodinated antibody
`VDS
`Ig
`pg
`mg
`
`24.5
`30.3
`300.8
`300.8
`722
`722
`924.4-
`1849t
`
`1.2
`1.5
`11.06
`11.06
`33.4
`33.4
`20.9-
`41.8t
`
`*protein-associated radioactivity.
`Idose given was within this estimated range-see Discussion.
`
`IMMUNOGEN 2297, pg. 3
`Phigenix v. Immunogen
`IPR2014-00676
`
`

`

`38
`
`C.H.J. FORD et al.
`
`Case
`
`Origin of primary
`
`1
`
`2
`
`3
`
`4
`
`5
`
`6
`
`7
`
`8
`
`Colon, Duke's C, well-
`differentiated
`adenocarcinoma
`
`Ovarian, FIGO IV
`mucinous
`cystadenocarcinoma
`
`Ovarian, FIGO III
`moderately well-
`differentiated
`adenocarcinoma
`Ovarian, FIGO
`III-IV
`adenocarcinoma
`
`Ovarian, FIGO III
`papillary
`cystadenocarcinoma
`
`Recto-sigmoid, Duke's
`C, well-differentiated
`adenocarcinoma
`
`Rectum, Duke's C,
`mucinous
`adenocarcinoma
`Caecum, Duke's C,
`adenocarcinoma
`
`*See text for details.
`
`Table II Summary of clinical localisation data
`
`Summary of disease
`at entry
`
`Widespread intra
`pulmonary metastases;
`pelvic and hepatic
`metastases
`Palpable abdomino-
`pelvic mass;
`left axillary
`nodes; left
`cervical nodes
`Mass in right
`groin; large
`left pelvic mass
`
`Left malignant
`pleural effusion;
`malignant peritoneal
`seedlings
`Pelvic recurrence
`
`Extensive pelvic mass
`(biopsy proven
`adenocarcinoma)
`and central
`palpable
`abdominal masses
`Perineal recurrence
`biopsy proven
`
`Pre-treatment
`Scan localisation
`serum CEA
`level (ngml-1) findings
`
`>3,550
`
`375
`
`29
`
`23
`
`32
`
`13
`
`Uniform liver (4 h);
`two small abdominal
`areas (48 h)
`
`Liver (4 h; 24 h);
`left axillar;
`right kidney;
`abdomino-pelvic
`mass (48 h)
`Central lower
`abdomen (48 h)
`
`Left chest (48 h);
`scattered areas
`in abdomen
`and pelvis (48 h)
`Medial to upper
`part of stomach
`scattered abdominal
`areas (48 h)
`Central abdominal
`and pelvic
`localisation (48 h)
`
`43
`
`*No localisation
`
`Retroperitoneal tumour,
`biopsy proven
`
`749
`
`*No localisation
`
`of 13 ng ml-1, one day after receiving 33.4mg VDS-
`Ig conjugate this level fell to 22ngml-'. Five days
`later the CEA level began to increase. In the other
`patients no change was observed.
`None of the patients had immediate or delayed
`hypersensitivity reactions to normal sheep Ig, either
`before the first or second dose of conjugate, nor any
`to the conjugate. The period between
`reaction
`conjugation and administration of the localising
`dose was 6-20 days; between conjugation and
`administration of escalated dose was 3-25 days, and
`
`the time between localising and escalated doses was
`4-54 days. Patients 7 and 8 received unconjugated
`antibody 8 and 6 days respectively before they
`received conjugate.
`In none of the 8 patients was there any toxicity
`or derangement of biochemical,
`renal
`or liver
`function which had not been present at entry into
`the investigation and which could be attributed to
`of conjugate.
`the administration
`Liver function
`became increasingly abnormal during follow-up in
`patient 3. Patient 6 developed obstructive jaundice
`
`IMMUNOGEN 2297, pg. 4
`Phigenix v. Immunogen
`IPR2014-00676
`
`

`

`A VINDESINE ANTI-CEA CONJUGATE IN VIVO
`
`39
`
`48 h abdominal subtraction scan for Patient 2, showing accumulation of isotope in the right kidney
`Figure 1
`(k) and in the abdomino-pelvic tumour mass (-+). Accumulation of isotope, in the form of free iodide, can be
`seen in the stomach at the top of the scan.
`
`48 h abdominal subtraction scan for Patient 6, showing accumulation of isotope in the pelvic
`Figure 2
`tumour mass (p) and in the central abdominal masses (-+). Accumulation of isotope, in the form of free iodide,
`can be seen in the stomach at the top of the scan.
`
`IMMUNOGEN 2297, pg. 5
`Phigenix v. Immunogen
`IPR2014-00676
`
`

`

`40
`
`C.H.J. FORD et al.
`
`and deranged liver function between the first and
`second administration of conjugate which was due
`to enlarged metastatic lymph nodes in the porta
`para-aortic
`and
`hepatis
`Progressive
`areas.
`gastrointestinal obstruction was noted in patients 1
`and 3 and was attributable to neoplastic adhesions
`which were confirmed at laparotomy for patient 1.
`Progressive pelvic recurrence was noted in patient 5
`during the study. Five of the patients were anaemic
`(3, 4, 5, 7 and 8) and only in the last patient was
`this progressive. Two patients (4 and 8) also had a
`short-lived thrombocytosis. In no case was the
`abnormal renal function in patients 1, 3, 4 and 5
`further impaired by administration of conjugate.
`peripheral sensory
`cis-platinum-related
`Also,
`the
`neuropathy
`patient
`in
`improved
`apparently
`4
`during the investigation.
`Three days after receiving radiolabelled conjugate
`patient 1 underwent sigmoidoscopy and biopsies of
`tumour and normal rectal tissue were obtained.
`These were weighed and the radioactivity measured
`in a y-counter. The ratio of normal:tumour (N:T)
`counts was 1:1.2. The tissues were then macerated
`with a scalpel blade, weighed, washed 3 x with
`RPMI 1640 and the counts remeasured. The N:T
`after
`ratio
`One
`week
`receiving
`1:4.6.
`was
`radiolabelled
`conjugate
`this
`had
`patient
`a
`laparotomy and was found to have numerous
`adhesions and liver secondaries. A colostomy was
`performed and biopsies taken of "normal" and
`metastatic
`hepatic
`tissue.
`Following
`the same
`procedure as before the N:T ratio was 1:0.6.
`
`Discussion
`
`One of the aims of this study was to determine
`"3'1-labelled VDS-
`whether radio-localisation
`of
`anti-CEA could be achieved in human tumours.
`This was clearly demonstrated in 4/8 patients (2, 3,
`4 & 6). Overall, the scanning results for Patient 1
`were also consistent with localisation. However, the
`uniform uptake in the liver at 4 h may have been
`secondaries,
`alternatively,
`due to
`liver
`or,
`to
`deposition of anti-CEA/CEA immune complexes
`(pre-treatment CEA level of >3,550ngml-1). The
`latter possibility is strengthened by the N:T ratio of
`radioactivity in the liver of 1:0.6. The two isolated
`showing
`abdominal
`localisation
`48 h
`areas
`at
`equated with the
`neoplastic adhesions
`seen
`at
`laparotomy. The pelvic and pulmonary metastases
`did not show localisation. In patient 5 localisation
`was equivocal.
`given
`Patients
`7 and 8
`were
`conjugate first, then scanned with radiolabelled
`antibody,
`unconjugated
`showed
`and
`neither
`convincing localisation. There are a number of
`Firstly,
`possible explanations
`for
`this.
`that
`the
`
`patients' tumours were not producing CEA. The
`pretreatment
`serum CEA levels
`43
`and
`of
`749 ng ml-' would argue against this. Secondly,
`avascularity could have reduced access
`of the
`conjugate to the tumour, resulting in false negativity
`suggested by others
`al.,
`(Dykes
`1980).
`as
`et
`However, we favour the third possibility which is
`that the CEA binding sites had been saturated by
`the administration of conjugate prior to receiving
`the radiolocalising dose. Both patients received up
`to 42mg of conjugated Ig 8 and 6 days respectively
`before their radiolocalising dose (Table 1).
`Our results suggest that drug conjugation has
`activity of the anti-CEA
`destroyed neither
`the
`antibody in vitro, nor its ability to localise in vivo.
`Most of the antibody activity was retained after
`conjugation as demonstrated by ELISA and the
`doses of conjugate required for localisation were
`similar to doses of unconjugated antibody reported
`by others (Goldenberg et al., 1978a; Dykes et al.,
`1980; Mach et al., 1980). Furthermore, there was an
`N:T radioactivity ratio of 1:4.6 at 3 days in a biopsy
`of the colonic tumour from patient 1.
`There was no obvious toxicity or hypersensitivity
`attributable
`administration
`of
`either
`to
`radiolocalising or escalated doses of conjugate in
`any of the eight patients. There were abnormalities,
`e.g. in liver function, during the course of the study,
`but none could be directly
`attributed
`the
`to
`conjugate. Most could be explained by the fact that
`patients
`the
`disease
`had
`advanced
`had
`and
`previously undertaken several different treatment
`programmes. The maximum dose of conjugated
`drug we administered was 0.9-1.8 mg on a single
`occasion, which is
`less than
`the
`conventional
`therapeutic dose of vindesine of 3-4 mg m 2 every 1-
`2 weeks (Yap et al., 1981; Cobleigh et al., 1981), or
`4-5mgm-' every 2 weeks (Valdivieso et al., 1981b).
`However, since the conjugate had been shown to be
`25 times as potent as free VDS against lung
`-%
`cancer cells in vitro (Johnson et al., 1981; Rowland
`et al., 1982a), we felt justified in taking a cautious
`approach when investigating it in patients.
`Overall there was no significant decrease
`in
`circulating CEA levels due to the administration of
`either dose of conjugate. Whilst Patients 3 and 6 did
`show decreases (31-17ngml-1 and 37-22ngml-1
`respectively),
`fluctuations
`similar
`these were
`to
`noted at other times and did not appear to be
`treatment related.
`have
`Whilst
`clearly
`demonstrated
`we
`the
`feasibility of this approach a number of problems
`were noted. The most important was aggregation of
`Batches III and IV, forcing us to change our plan of
`investigation. For Batch III 80% relative anti-CEA
`activity was used to
`calculate
`the amount of
`conjugate given. However, for Batch IV it was not
`
`IMMUNOGEN 2297, pg. 6
`Phigenix v. Immunogen
`IPR2014-00676
`
`

`

`A VINDESINE ANTI-CEA CONJUGATE IN VIVO
`
`41
`
`possible to do this. We estimate the dose given was
`5O-100%
`within
`the
`The
`range.
`for
`reason
`aggregation
`unexplained
`remains
`present. A
`at
`possible
`factor
`have
`been
`the
`may
`higher
`conjugation ratio of 11:1 achieved with Batch IV. A
`single batch of conjugate was not used for the entire
`study in order
`to minimise the time between
`conjugation
`administration
`and
`of
`both
`radiolocalising and escalated doses, since a loss of
`anti-CEA activity of the conjugate had been noted
`in vitro after storage at 4°C for more than 31 days
`(Johnson et al., 1981). The longest period between
`conjugation and administration of conjugate in the
`present study was 25 days.
`CEA was chosen as a model for this investigation
`because, by immunocytochemistry,
`it has been
`shown to
`be
`expressed
`by 62% of
`colonic
`al.,
`carcinomas (Goldenberg et
`1978b), 62% of
`gastric carcinomas (Lee
`al.,
`1978), 63% of
`et
`invasive cancers of the cervix (Van Nagell et al.,
`1979), and 82% of lung cancers (Ford et al., 1981).
`Also, 100% of primary and 67% of metastatic
`ovarian carcinoma sites have been shown to localise
`anti-CEA antibodies in vivo (Van Nagell et al., 1980)
`and successes
`have
`achieved
`been
`with
`other
`tumours (Goldenberg et al., 1978a; Dykes et al.,
`1980; Mach et al., 1980), although, as in this study,
`not all tumour sites in a patient and not all patients
`have shown localisation. CEA, therefore,
`is
`a
`potential target applicable to a variety of human
`cancers.
`
`Vindesine
`is
`active
`in
`cell
`small
`anaplastic
`carcinoma of the lung (Osterlind
`al.,
`et
`1981),
`colorectal
`(Valdivieso
`1981a) and breast
`al.,
`et
`carcinomas (Yap et al., 1981; Cobleigh et al., 1981).
`Since it is feasible to make conjugates of vindesine
`with
`antibody,
`VDS-anti-CEA
`conjugates
`are
`candidates
`clinical
`for
`evaluation
`possible
`of
`therapeutic
`effect.
`Questions
`that
`also
`require
`resolution
`and
`which
`the
`subject
`of
`are
`investigation are whether the conjugate is stable in
`vivo and whether
`administration
`of conjugate
`actually results in increased tissue levels of drug in
`the target tissue. We are exploring this potential
`further in vitro and in vivo using polyclonal and
`monoclonal VDS-antibody conjugates (Rowland et
`al., 1982ab). Clinical use of monoclonal conjugates
`should be acceptable in view of the reports of
`successful administration of monoclonal antibodies
`to patients (Nadler et al., 1980; Miller & Levy,
`1981).
`
`We thank Mr. W. Smith and Mrs. C.H. Marsden for help
`with the preparation of the conjugates; Mr. J. Griffin for
`the preparation of CEA, and the following physicians for
`access to their patients: Professor F. Ashton, Mr. P.
`McMaster, Mr. J. Fielding, Mr. W. Bond and Dr. A.
`Banks. We are indebted to Mrs. Z. Drolc for allowing us
`access to the computerised subtraction facilities in the
`of Nuclear
`Department
`Medicine,
`Elizabeth
`Queen
`Hospital, and to Mr. V. Trend for performing the
`bacteriology tests.
`
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