This material may be protected by Copyright law (Title 17 U.S. Code)
`
`Merck 2022
`TWi v Merck
`IPR2023-00049
`
`

`

`DISPOSITION OF MITOXANTRONE IN CANCER PATIENTS
`
`HO
`
`ql
`
`*
`
`*
`
`H
`
`NCHjCHaNCH2CH20H
`
`- 2HC!
`
`Hf
`
`*
`x
`NEHZCHANCHZCHSOH
`HO
`Chart 1. Chemical structure of mitoxantrone (1,4-dihydroxy-5,8-bis|{{2-{(2-hy-
`droxyethyl)amino}ethy!}amino}}-9,10-anthracenedione dihydrochloride). *, positions
`of “C radiolabels.
`
`um Fluoroporefilter (Millipore Corp., Bedford, MA) prior to use. Aqueous
`solvents for HPLC use werefiltered through a 0.45-um cellulose acetate
`filter prior to use. Ammonium acetate (HPLC grade) was obtained from
`Fisher Scientific Co. (Fair Lawn, NY). t-Ascorbic acid was obtained from
`GrandIsland Biological Co. (Grand Island, NY). Human albumin and y-
`globulin were obtained from Sigma ChemicalCo.(St. Louis, MO), and 3
`N methanolic-HCl was obtained from Supelco (Bellefonte, PA).
`HPLC Sample Cleanup Procedure. A VAC-ELUT system equipped
`with BOND-ELUT 1-ml Cig cartridges (Analytichem International, Harbor
`City, CA) was used for sample cleanup. One- to 2-mi plasma samples
`were passed through a cartridge which was sequentially preconditioned
`by washing with 10 ml of methanol and 5 ml of H2O. After the plasma
`had passed throughthe cartridge, it was washed with 5 ml of H2O, and
`300 ul of 0.5 nN methanolic-HCl were used to elute the drug. The eluate
`wascollected, vortexed, and kept at —20° C for HPLC analysis.
`HPLC Apparatus. HPLC was performed with an apparatus consisting
`of a Model 660 solvent programmer, 2 Model 6000A solvent delivery
`systems, a Mode! U6K injector, a Mode! 440 detector (Waters Associ-
`ates, Milford, MA), and a Model A-25 recorder (Varian, Palo Alto, CA). A
`Waters Associates C1s-uBondapak (3.9-mm inside diameter x 30 cm)
`reversed-phase column preceded by a 7-cm x 2.1-mm (inside diameter)
`guard column packed with CO:PELL ODS (Whatman Inc., Clifton, NJ)
`wasused forall analyses. The guard column packing was changed every
`2 weeks or whenever there wassignificant back pressure buildup.
`HPLC Assay Chromatographic Conditions. Mitoxantrone waseluted
`isocratically at ambient temperature with a solvent composition of 25%
`CH3CN and 75% ammonium acetate (0.2 m, pH 4.0) at a flow rate of 1.5
`ml/min. Mitoxantrone was detected at 658 nm using a Waters Associates
`Model 440 fixed-wavelength detector.
`HPLC Assay Quantitation. Quantitation of mitoxantrone was done by
`the external standard method of analysis. Plasma standard curves were
`obtained by plotting the resulting peak heights against the known con-
`centration of standards added to the plasma samples.
`Tissue Preparation for Quantitation of “C-Labeled Mitoxantrone
`Equivalents. Plasma, urine, and saliva samples were prepared in a
`similar way. In each case, 0.5 ml of sample was added to 0.5 or 1 ml of
`solubilizer plus 10 ml of Aquasol, mixed, and counted in a scintillation
`counterfor 10 min. Whole bone marrow, whole blood, and RBC (0.1 to
`0.2 ml) were added to 0.5 ml of concentrated acetic acid in a scintillation
`vial and mixed until RBC were solubilized; 0.5 mi of hydrogen peroxide
`(30%) was added, and the solution was mixed well and heated to 60° C
`for 15 min until decolored. The samples were then cooled at room
`temperature, each added to 10 ml of Aquasol, and counted for 10 min
`in the scintillation counter. Each sample wasthen runin at least duplicate.
`Two ml of water were added to each 1 g of fecal sample and sonicated.
`One mi of the fecal mixture was then added to 10 ml of solubilizer in a
`scintillation vial and heated at 60° C for 30 min. Then 0.1 ml of the
`digested material was added to 10-mi Ready-Solv-HP, mixed well, and
`counted for 10 min in the scintillation counter.
`All samples were run in at least duplicate. Organ samples were
`weighed and oxidized using a Packard Tri-Carb Mode! B306 sample
`oxidizer. The “CO2 formed during oxidation was trapped in 4 ml of
`Carbosorb (Packard Instrument Co.). Twelve ml of Beta-Phase (West
`
`Chem Co.) scintillation fluid were added. The resultant cocktail was
`counted on a Beckman LS100 liquid scintillation counter for 10 min. The
`data obtained were expressed as ng/g, wet weight, of tissue.
`Each tumor biopsy was disaggregated into a single cell suspension.
`The cell suspension was then washed and counted. Aliquots of the cell
`suspension were added to a glassscintillation vial containing 10 ml of
`Aquasol, and were then counted for 10 min in a Beckman LS230
`scintillation counter. All counts were corrected for background and
`counting efficiency. The data were expressed as ng/million cells.
`The solubilizer used for these experiments was BTS-450 from Beck-
`man, and the Aquaso! was type NEF-934 from New England Nuclear.
`The Ready-Solv-HP wasalso obtained from Beckman. The scintillation
`counter was used at Channel B “‘C with a gait of 260 and a preset error
`of 0.2%.
`Plasma Mitoxantrone Pharmacokinetic Analysis Techniques. The
`mitoxantrone concentration versus time plasma data werefit to the
`multiexponential equation
`
`C = Ae“ + Be-™ + Cem
`
`using BMDP 3R, a nonlinear regression computer program. A weighting
`function of 1/y? was used in order to fit the later time points. Initial
`parameter estimates are required for nonlinear regression and were
`obtained using CSTRIP. F-Tests were carried out to determine the
`number of compartments needed. Since there were insufficient data
`points during the infusion period to adequately define the area under this
`portion of the curve, for the pharmacokinetic analysis, the infusion data
`were adjusted to represent the equivalent of a bolus injection, using the
`mathematical equations described by Gibaldi and Perrier (7).
`
`RESULTS
`
`Patient Demography and Diagnosis. Eight patients were
`enrolled into this study. The demographics and diagnostic char-
`acteristics of the patient population are presented in Table 1.
`There were 3 male and 5 female patients with a mean age of
`57.9 years (range, 25 to 74). The mean body surface area was
`1.61 sq m (range, 1.37 to 1.90), and the mean weight was 60.8
`kg (range, 45.7 to 77.0). All patients enrolled in the study had
`advanced,soft-tissue primary disease (4 head and neck cancers,
`2 renal cancers, 1 ovarian cancer, and 1 melanoma). None had
`primary bone involvement or leukemia, although one patient
`(Patient 2) had skeletal metastases. Prior to pharmacokinetic
`study, 3 patients (Patients 5, 6, and 7) had received 2, 2, and 12
`doses of 12 mg mitoxantrone/sq m, respectively. These 3 pa-
`tients were all receiving mitoxantrone once every 3 weeks.All
`other patients were studied during the first dose administration.
`Plasma and Blood Cellular Drug Concentrations. The
`plasma mitoxantrone (HPLC) and ['*C]mitoxantrone (expressed
`as mitoxantrone equivalents ng/ml) concentrations were ob-
`tained in 7 of the 8 patients. No plasma samples were taken
`from Patient 8 because ofdifficulty with adequate peripheral vein
`access. Plasma mitoxantrone and ['*C]mitoxantrone concentra-
`tions decreased rapidly during the first 1 to 2 h after infusion
`completion. Thereafter, the decrease was slower. During the
`initial time period, plasma radioactivity concentrations were ap-
`proximately equal to plasma mitoxantrone concentrations mea-
`sured by HPLC. The decrease in plasmaradioactivity concentra-
`tions was best described by dividing the patients into 2 groups.
`In Group 1 (Patients 1, 2, and 4), plasma radioactivity concentra-
`tions approximated plasma mitoxantrone concentrations for 5 to
`20 min. At the later time periods, plasma radioactivity concentra-
`tions decreased less rapidly than plasma mitoxantrone concen-
`
`CANCER RESEARCH VOL. 45 APRIL 1985
`1880
`
`

`

`DISPOSITION OF MITOXANTRONE IN CANCER PATIENTS
`
`4
`
`Ovarian adenocarcinoma
`
`166
`
`475
`
`1.42
`
`67
`
`F/C
`
`18
`
`No
`
`Table 1
`Demographic and disease status characteristics of patients entered into pharmacokinetic study of mitoxantrone
`Total
`Prior mitox-
`dose
`Sex/
`Age
`BSA
`Wt
`Height
`Site of metastasis
`Disease type
`Patient
`
`
`
`(cm) (Mxm)—(yr)(kg) race (mg) antrone
`
`
`1
`Renal leiomyosarcoma
`None
`156
`51.5
`142
`58
`FIC?
`17
`No
`2
`Renal adenocarcinoma
`Bone (multiple)
`168
`49.8
`137
`40
`F/C
`16
`No
`3
`H and N squamouscell
`Lung; submandibular
`176
`73.5
`190
`74
`M/C
`23
`No
`lymph node
`Cul-de-sac; supracia-
`vicular lymph node
`Lung
`
`5
`
`172
`
`Salivary gland adenocarci-
`noma
`Lung
`Tongue squamouscell
`6
`Yes(2)*
`21
`M/C
`71
`1.75
`69.0
`164
`Lung
`Submandibular gland adeno-
`7
`
`
`
`
`
`175 FIC=22770 1.80 58 Yes (12)*
`cystic carcinoma
`Melanoma
`
`727
`
`185
`
`70
`
`M/C*
`
`22
`
`Yes (2)*
`
`8
`
`Skin (multiple)
`
`154
`
`45.7
`
`1.40
`
`25
`
`F/C
`
`18
`
`No
`
`Mean
`Minimum
`Maximum
`
`1688
`156
`172
`
`608
`457
`770
`
`1.61
`137
`19
`
`57.9
`25
`74
`
`19.6
`16
`23
`
`* C, Caucasian of European ancestry; C*, Mexican; *, number of prior courses of mitoxantrone.
`
`10
`
`100
`
`e@prasmalclaiTROXANTRONE EQUIVALENTS
`© PLASMA HPLC-MITOXANTRONE
`o RBC4c]MITOXANTRONE EQUIVALENTS
`
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`2
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`z
`bq ps
`2
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`<
`‘
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`wee eee
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`CONCENTRATION(g/m!)
`o eeeeeeesLe,
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`e prasma|'4qmitoxaNTRONE EQUIVALENTS
`O PLASMA HPLC-MITOXANTRONE
`corse ['4duitoxaNTRONE EQUIVALENTS
`
`0
`
`20
`
`40
`TIME (HOURS)
`
`60
`
`80
`
`100
`
`TIME (HOURS)
`
`Chart 2. Concentration time data for mitoxantrone measured by HPLC and '*C-
`radioactivity in plasma and in RBC.['*C]Mitoxantrone (specific activity, 8.85 uCi/
`mg) was administered i.v. over a 30-min period in a dose of 12 mg/sq m to Patient
`4 (Tables 1 and 2).
`
`Chart 3. Concentration time data for mitoxantrone measured by HPLC and '4C-
`radioactivity in plasma and in RBC. ['C]Mitoxantrone (specific activity, 8.85 uCi/
`mg) was administered i.v. over a 30-min period in a dose of 12 mg/sq m to Patient
`6 (Tables 1 and 2).
`
`trations (e.g., Chart 2). The difference persisted for the remainder
`of the study, approaching a 5-fold difference when the plasma
`mitoxantrone concentration had reached 2 ng/ml. In the other
`group (Group 2) of patients (Patients 3, 5, 6, and 7), the decline
`with time after administration of plasma radioactivity concentra-
`tions paralleled the decline of the plasma mitoxantrone concen-
`trations with little difference between the 2 (e.g., Chart 3).
`The ['*C]mitoxantrone content in blood FE was also deter-
`mined following separation of plasma. The concentration of
`radioactivity in the FE was consistently greater than that in the
`plasma. The FE/plasmaratio of radioactivity ranged from 2/1 to
`10/1 at various sampling times after mitoxantrone administration
`(e.g., Charts 2 and 3).
`Plasma Pharmacokinetic Parameters.In 5 of the 7 evaluable
`patients, the plasma disappearance of mitoxantrone concentra-
`
`tions measured by HPLC were best described by a 3-compart-
`ment model. The mean pharmacokinetic parameters for these 5
`patients are shown in Table 2. Because the data from the
`remaining 2 patients (Patients 4 and 5; Tables 1 and 2) were
`adequately described by a 2-compartment model, they have not
`been included in this analysis. In Patient 4, the terminal-phase
`(y) plasmahalf-life of mitoxantrone wassimilar to that determined
`for Patients 1, 2, 3, 6, and 7, but her t,, was an average of the
`a- and B-phases of the 3-compartmentfit. In Patient 5, the
`plasma concentrations of mitoxantrone were below the sensitiv-
`ity level of our HPLC assay(i.e., <1 ng/ml plasma) within 7 h of
`drug administration. Thus,
`it was not possible to determine
`accurately a plasma terminal elimination phase in this patient.
`in the 5 patients shown in Table 2, the initial phase of the
`decrease in plasma mitoxantrone concentrations was rapid
`
`CANCER RESEARCH VOL. 45 APRIL 1985
`
`1881
`
`€
`

`

`DISPOSITION OF MITOXANTRONE IN CANCER PATIENTS
`
`Table 2
`Individual pharmacokinetic parameters
`12-mg/sq m dose
`
`Patient
`1
`2
`3
`6
`7
`Mean+SD
`
`a (Hh)
`8.7
`9.6
`8.5
`3.7
`6.2
`7.342.
`
`B(h")
`0.67
`0.67
`0.72
`0.45
`0.93
`0.694017
`
`¥ (bh)
`0.03
`0.01
`0.01
`0.01
`0.028
`0.02+0.01
`
`t* (h)
`0.078
`0.072
`0.082
`0.187
`0.112
`0.10640.048
`
`Ch (liters/
`min/sqm)
`t(h)
`t* (h)
`0.94
`20.8
`1.04
`0.65
`51.7
`1.05
`0.30
`69.3
`0.95
`0.47
`46.2
`1.54
`0.50
`25.0
`0.75
`1.0740.29 42641996 057+0.24
`
` Cl_ (ml/
`—min/sq m)
`85.8
`55.4
`18.5
`27.1
`38.6
`45422
`
`V. (liters/
`sqm)
`9.7
`12.4
`3.9
`24.6
`10.3
`12184762
`
`Vey (liters/sq
`m)
`1695
`2930
`1797
`1878
`1072
`1875+670
`
`Table 3
`
`Table 4
`
`Mean urinary recovery of mitoxantrone and [‘“C]mitoxantrone
`%of administered dose
`
`Time (h)
`
`0-4
`4-8
`8-16
`16-24
`
`0-24
`24-48
`48-72
`72-96
`96-120
`Mean total recovery (0
`to 120 h)
`Range
`
`No.of
`samples
`6
`6
`5
`5
`
`{"4C]Mitoxantrone
`29
`1.2
`0.6
`0.6
`
`No. of
`samples Mitoxantrone
`6
`3.7
`6
`1.2
`5
`0.4
`5
`0.5
`
`7
`7
`7
`7
`4
`
`5.6
`1.4
`1.0
`0.8
`0.9
`10.1
`
`7
`6
`7
`6
`4
`
`5.9
`0.5
`0.3
`0.3
`0.2
`65
`
`6.2-23.5
`
`5.2-7.9
`
`Fecal [“*C]mitoxantrone recovery
`No. of bowel
`movements
`;
`1
`7
`4
`5
`4
`1
`
`Patient
`>
`3
`4
`5
`6
`7
`8
`
`% of dose recovered
`(0-120 h)
`ve
`0
`15.4
`13.6
`20.0
`24.8
`2.2
`
`Mean %" of administered
`dose (0-120 h)
`13.6-24.8
`Range
`* Includes onty those patients who had more than one bowel movement.
`
`18.3
`
`Table 5
`Saliva [‘‘C]mitoxantrone concentrations in Patient 8
`Time postinfusion
`Concentration equivalents
`(h)
`(ng/ml)
`0.25
`3.8
`oe
`1.00
`2.00
`rer
`8.00
`12.00
`oe
`96.00
`120.00
`
`25
`43
`a8
`75
`11.4
`oe
`40
`3.6
`
`(meanty, = 0.1 h). The second phase (t,,.) was somewhat longer
`with a mean half-life of 1.1 h. The mean terminal half-life (ty.) was
`42.6 h. The mean apparent volume of the central compartment
`(V.) was 12.2 liters/sq m, while the mean apparent volume of
`distribution (Va,) was 1875liters/sq m. The mean plasmaclear-
`ance(C/;) was 0.57liters/min/sq m, and the mean renal clearance
`(Cla) was 45 mli/min/sq m, which represents 12%of the plasma
`clearance.
`Urinary Recovery of Mitoxantrone and [“C]Mitoxantrone.
`The mean urinary recoveries of mitoxantrone and ["*C]mitoxan-
`trone at intervals of 4 to 120 h following drug administration are
`shownin Table 3. Only 6.5% (range, 5.2 to 7.9%) of the total
`Table 6
`mitoxantrone dose was recovered as unchanged drug over 5
`["*C]Mitoxantrone equivalents in biopsy and autopsy specimens
`days (120 h). The majority (90%) of drug was recovered during
`Time after|Concentration®
`the first 24 h with the first 4 h contributing the largest quantity
`["*C]mitoxan-
`equivalents
`(62%). The HPLC chromatograms of patient urine samples
`trone
`(ngfliter x 10°
`showed consistently up to 3 more polar metabolite peaks than
`(h)
`cells)
`that which identified the parent compound.
`6
`Meanrecovery of '*C-labeled material during the 120-h collec-
`tion period was 10.1%(range, 6.2 to 23.5%) of the administered
`dose. Fifty-five % was recovered during the first 24 h. The
`recovery of ‘C-labeled material for 7 of 8 patients (excluding
`Patient 2) ranged from 0.8- to 1.6-fold the parent compound (i.e.,
`mitoxantrone) recovery. Patient 2 excreted 23%of the adminis-
`tered dose as ['*C]mitoxantrone of which only 7.9% (as a
`percentage of dose administration) proved to be parent com-
`pound.
`Fecal Recovery of Mitoxantrone and [‘*C]Mitoxantrone.
`Feces werecollected from 7 patients during the 5 daysof study.
`Dueto the health status and dietary restrictions of the patients,
`fecal output was variable, ranging from 0 to 9 collections over
`120 h (Table 4). In those patients who had morethan one bowel
`
`Patient
`2
`
`3
`
`4
`
`8
`
`Tissue
`Bone marrow
`Whole
`RBC only
`WBC only
`Squamouscell, tumor nodule,
`neck
`Adenocarcinoma, supracla-
`vicular lymph node
`Metastatic melanoma nodule,
`wrist
`
`* 1x 10° cells = ~1 mg.
`
`0.06
`0.05
`1.13
`0.06
`
`1.32
`
`0.03
`
`§.75
`
`6
`
`22.75
`
`movement during the 5-day collection period, the mean percent-
`age of recovery of ‘C-labeled material was 18.3% (range, 13.6
`to 24.8%) of the administered dose.
`[“C]Mitoxantrone in Saliva. Saliva was collected from pa-
`tients to determine if radiolabeled drug was secreted via this
`
`CANCER RESEARCH VOL. 45 APRIL 1985
`1882
`
`

`

`2,000,000
`
`500,000
`
`
`
`TOTALNG/ORGAN
`
`$0,000
`
`25,000 8000
`
`DISPOSITION OF MITOXANTRONE IN CANCER PATIENTS
`
`pathway. Only 1 to 3 samples were collected from mostpatients;
`however, Patient 8 provided several additional specimens
`throughout the 120-h collection period. Drug recoveries from
`saliva obtained from Patient 8 are shownin Table 5.
`Tissue Concentrations of [“C]Mitoxantrone Equivalents.
`As shownin Table 6, tumor biopsies were obtained 5 to 22 h
`following ["*C]mitoxantrone administration in 3 patients (Patients
`3, 4, and 8). Only pg quantities of mitoxantrone equivalents per
`10° cells were recovered from a squamouscell tumor nodule
`(Patient 3) and a melanoma nodule (Patient 8) in 2 nonresponding
`patients. Of interest was the observation that metastatic tumor
`tissue from one of these patients (Patient 8) had proven ex-
`tremely sensitive in vitro to mitoxantrone (i.e., less than 1%
`survival of tumor colony-forming units at 10 ng/ml for a 1-h
`exposure) as measured by a human tumor clonogenic assay
`(14). A metastatic supraclavicular lymph node aspirated 6 h after
`mitoxantronein a patient with ovarian cancer(Patient 4) revealed
`considerably higher amounts of ['*C]mitoxantrone equivalents
`(i.e., >1 ng/10® tumor cells). Although this patient was evaluated
`as a nonresponder to mitoxantrone, the lymph node disease did
`decrease in volume by greater than 25% during therapy.
`A bone marrow aspirate obtained 6 h after mitoxantrone
`administration to a patient (Patient 2) with skeletal metastases
`secondary to a renal cell cancer showed approximately a 20-fold
`greater concentration of '*C-drug equivalents in the WBC versus
`RBCfraction (as separated by a Ficoll-Hypaque gradient) (Table
`6). The ratio of whole marrow to plasma (adjusted for a hema-
`tocrit of 30%) was approximately 8/1. This patient experienced
`life-threatening leukopenia (i.e., neutrophil count <500/cu mm)
`10 days following mitoxantrone therapy.
`Organ specimens were obtained from one patient (Patient 2)
`who died of progressive kidney cancer 35 days after mitoxan-
`trone administration. Even at 35 days, the liver, pancreas, thy-
`roid, spleen, and heart contained relatively high mitoxantrone
`equivalents per g of tissue (wet weight) (Chart 4). On the basis
`of ['*C]mitoxantrone distribution per whole organ, the liver con-
`tained the highest amountof drug followed by the bone marrow,
`heart, lungs, spleen, kidney, and thyroid glands in that order
`(Chart 5). Adding the total amounts of mitoxantrone retained in
`each of these 7 organs as much as 15%of the administered
`dose could be accounted for in these tissues at 35 days.
`
`DISCUSSION
`
`We have shownthat the plasma disappearance of mitoxan-
`trone measured by HPLC can be described in the majority of
`
`
`
`0:
`
`Chart 4. ['*C]Mitoxantrone equivalents in ng per g of tissue (wet weight) ob-
`tained at autopsy 35 days after iv. administration in a dose of 12 mg/sq m to
`Patient 2 (Tables 1 and 2).
`
`Chart 5. [*C]Mitoxantrone equivalents in ng per whole organ (wet weight)
`obtained at autopsy 35 days after i.v. administration in a dose of 12 mg/sq m in
`Patient 2 (Tables 1 and 2).
`
`patients by a 3-compartment model with a prolonged terminal
`elimination phase half-life of approximately 43 h. Previous inves-
`tigations have reported either shorter (8) or similar (15, 16)
`durations of this important pharmacokinetic parameter. Our
`highly sensitive HPLC assay (12) has allowed us to measure
`mitoxantrone plasma concentrations for up to 72 h after drug
`administration, and thus accurately determine the duration of the
`terminal elimination phase half-life. While the estimate of this
`half-life (i.e., range of 20.8 to 69.3 h)is longer than that reported
`by others(8),it is likely that the true elimination half-life is much
`longer based on the body content of drug 35 days after dosing.
`These data provide a pharmacological rationale for the use of
`mitoxantrone on an intermittent dosing schedule.
`Mitoxantrone appears to distribute into a deep tissue com-
`partment from which it is slowly released as evidenced byits
`prolonged plasma terminal-phase half-life, extremely large vol-
`ume of distribution (Vo), and the relatively large amount of
`mitoxantrone (>15%of administered dose) which appeared to
`be retained in autopsy tissues 35 days after dosing. Even though
`mitoxantrone (and/or mitoxantrone-related material) may persist
`in the body for prolonged periods, repeat dosing at 3-week
`intervals for as many as 12 courses had no noticeable effect on
`the calculated pharmacokinetics.
`Considerable evidence exists to suggest that mitoxantrone
`undergoes extensive metabolism, probably in the liver. (a) The
`mean area under the plasma disappearance curvefor “C-labeled
`material wassignificantly greater than that determined by HPLC.
`However, in Patients 3, 5, 6, and 7 (Tables 1 and 2; Chart 3),
`the decline with time after administration of [‘*C]mitoxantrone
`plasma concentrations paralleled the decline of the plasma mi-
`toxantrone concentrations measured by HPLC withlittle differ-
`ence between them.It is possible that, in this group of patients,
`for unknown reasons,there was a lesser degree of metabolism
`
`CANCER RESEARCH VOL. 45 APRIL 1985
`1883
`
`

`

`DISPOSITION OF MITOXANTRONE IN CANCER PATIENTS
`
`REFERENCES
`
`depends on adequate drug uptake into target tissue. Drug
`disposition studies of this type can provide important insights
`when interpreting the results of Phase II clinical trials.
`
`than occurred in Patients 1, 2, and 4. There was no evidence of
`liver dysfunction in any of these patients which could have
`explained differences in the pattern of mitoxantrone plasma
`clearance. (b) The recovery of ‘C-labeled material in the 5-day
`urine collections wassignificantly greater than that of the HPLC-
`measured parent compound. (c) Urine HPLC chromatograms
`1. Aapro, M., and Alberts, D. S. Phase Il trial of mitoxantrone in head and neck
`revealed up to 3 polar metabolites, which appeared identical to
`cancer. Invest. N. Drugs, 2: 329-330, 1984.
`those observed previously in rat bile.* (¢) Ehningeretal. (5) have
`2. Aapro, M., Mackel, C., Alberts, D., and Woolfenden, J. Phase II cardiotoxicity
`study of mitoxantrone hydrochloride using exercise radionuctide evaluation of
`shownrecently, using an isolated perfused rat liver model and
`the left ventricular cardiac ejection fraction (LVEF). Proc. Am. Soc.Clin. Oncol.,
`1: 14, 1982.
`thin-layer chromatography assay techniques, that mitoxantrone
`is actively metabolized with up to 3 more polar compounds
`3. Alberts, D. S., Griffith, K. S., Goodman, G. E., Herman,T. S., and Murray,E.
`Phase|clinical trial of mitoxantrone: a new anthracenedione anticancer drug.
`appearing on the thin-layer plates. The identity and potential
`Cancer Chemother. Pharmacol., 5: 11-15, 1980.
`activity of these putative metabolites have not been determined
`4. Alberts, D. S., Peng, Y-M., and Davis, T. P. Comparative pharmacology of
`mitoxantrone and bisantrene. Clin. Pharmacol. Ther., 37: 199-200, 1982.
`due to their relatively small quantities and insensitivity of the gas
`5. Ehninger, G., Proksch, B., Hartmann, F., Garther, H-V., and Wilms, K. Mitox-
`chromatography-mass spectrometry techniques thus far applied
`antrone metabolism in the isolated perfused rat liver. Cancer Chemother.
`to these studies.
`Pharmacol. 12(1): 50-52, 1984.
`6. Estey, E. H., Keating, M. J., McCredie, K. B., Bodey, G. P., Freireich, E. J.
`The most important route of mitoxantrone elimination appears
`Phase II trial of dihydroxyanthracenedione in acute leukemia. Proc. Am. Asoc.
`to be fecal. Total drug-related material recovered in urine plus
`Cancer Res., 23: 113, 1982.
`7. Gibaldi, M., and Perrier, D. in: Pharmacokinetics. New York: Marcel Dekker,
`stool averaged 28% of the administered dose in 5 days; 10.1%
`Inc., 1975.
`in the urine (6.5% as mitoxantrone and an additional 3.6% as
`8. Neidhart, J., Stabus, A., Young, D., and Malspeis, L. Pharmacokinetic studies
`C-labeled material) and 18% in the feces. Because of the
`of dihydroxyanthracenedione (DHAD, NSC 301,739)with clinical correlations.
`Proc. Am. Assoc. Cancer Res., 22: 363, 1981.
`relatively low urinary excretion of mitoxantrone and its metabo-
`9. Neidhart, J. A., and Roach, R. W. A randomized study of mitoxantrone (M)
`lites, it is unlikely that the standard drug dose must be reduced
`and adriamycin (A) in breast cancer patients failing primary therapy. Proc. Am.
`Soc. Clin. Oncol., 1: 86, 1982.
`in the presence of compromised renal function. On the other
`10. Ostroy, F., Gams, R. A. An HPLC method for the quantitative determination of
`hand, since the drug appears to be metabolized in the liver,
`1,4-dihydroxy-5,8 bis((2+(2-hydroxyethyl) amino)ethyl)amino) 9,10-anthrace-
`future studies must be carried out to determine the effectofliver
`nedione (DHAD, Led. Labs., CL 232,315, NSC 301739)in serum. J. Liquid
`Chromatogr., 3: 637-644, 1980.
`dysfunction on the disposition and toxicity of mitoxantrone.
`11. Peng, Y-M., Davis, T. P., and Alberts, D. S. High performance liquid chroma-
`Indeed, Savaraj et al. (15) have shownthat patients treated with
`tography of a new anticancer drug, ADCA-physicochemical properties and
`mitoxantrone who had either abnormalliver function or a third
`pharmacokinetics.Life Sci., 29: 361-369, 1981.
`12. Peng, Y-M., Ormberg, D., Alberts, D. S., and Davis, T. P. Improved high-
`space appeared to havesignificantly prolonged terminal-phase
`performance liquid chromatography of the new antineoplastic agents bisan-
`plasmahalf-lives and decreased rates of total body clearance of
`trene and mitoxantrone. J. Chromatogr. Biomed. Appl., 233: 235-247, 1982.
`the parent compound.
`13. Reynolds, D. L., Sternson,L. A., and Repta,A.J. Clinical analysis for the anti-
`neoplastic
`agent
`1,4-dihydroxy-5,8-bis{ {{2-[(2-hydroxyethy!)amino]ethyl}-
`Thelimited salivary secretion data obtained in this study show
`amino}} 9,10-anthracenedione dihydrochloride (NSC 301739) in plasma. J.
`prolonged, but relatively low concentrations of mitoxantrone in
`Chromatogr., 222: 225-240, 1981.
`14. Salmon, S. E., Hamburger, A. W., Soehnien, B., Durie, B. G. M., Alberts, D.
`saliva for up to 120 h after drug administration. In that there
`S., and Moon,T. E. Quantitation of differential sensitivity of human tumor stem
`appeared to be mitoxantrone activity against a salivary gland
`Cells to anticancer drugs. N. Engl. J. Med., 298: 1321-1327, 1978.
`cancer in one Phase II clinical trial (1), these distribution data
`15. Savaraj, N., Lu, K., Manuel, V., and Loo, T. L. Pharmacology of mitoxantrone
`in cancer patients. Cancer Chemother. Pharmacol., 8: 113-117, 1982.
`provide a rationale for carrying out additional trials of mitoxan-
`16. Savaraj, N., Lu, K., Valdivieso, M., Burgess, M., Umsawadi, T., Benjamin, R.
`trone in the treatment of these relatively rare tumors. Because
`S., and Loo, T. L. Clinical kinetics of 1,4-dihydroxy-5,8-bis((2-(2-hydroxy-
`ethyl)amino) ethyl}amino)-9,10-anthracenedione. Clin. Pharmacol. Ther., 37:
`the concentrations of mitoxantronein saliva were quite low,it is
`312-316, 1982.
`unlikely that further monitoring of this elimination route would
`17. Sparano, B. M., Gordon,G., Hall, C., latropoulos, M. J., and Nobel, J. F. Safety
`prove useful in the evaluation of mitoxantrone pharmacokinetics.
`assessment of a new anticancer compound, mitoxantrone, in beagle dogs:
`comparison with doxorubicin II. Histologic and ultrastructural pathology. Can-
`The mitoxantrone tissue disposition data obtained in 4 of our
`cer Treat. Rep., 66: 1145-1158, 1982.
`patients suggest a direct relationship between the degree of
`18. Stuart-Harris, R. C., and Smith,
`|. E. Mitoxantrone: a phase II study in the
`treatment of patients with advanced breast carcinoma and other solid tumours.
`drug uptake into tumor or normal tissue cells and biological
`Cancer Chemother. Pharmacol., 8: 179-182, 1982.
`effect. Although metastatic melanoma cells obtained prior to
`19. Unverferth, D. V., Underferth, B. J., Balcerzak, S. P., and Neidhart, J. A.
`therapy in one of our patients had proven exquisitely sensitive
`Cardiac evaluation of mitoxantrone. Cancer Treat. Rep., 67: 343-350, 1983.
`20. Van Echo, D. A., Shulman, P. N., Ferrari, A., Budman, D., Markus, S. D., and
`to mitoxantrone in vitro (14), treatment was associated with
`Wierik, P. H. A phase II trial of mitoxantrone (DHAD, NSC301739)in adult
`progressive disease and only pg quantities of ‘C-labeled drug
`acute leukemia (AL). Proc. Soc. Clin. Oncol., 1: 132, 1982.
`equivalents per 1 x 10° cells were recovered approximately 23
`21. Von Hoff, D. D., Pollard, E., Kuhn, J., Murray, E., Coltman, C. A., Osbome,C.
`K. Phase ! clinical
`investigation of 1,4-dihydroxy-5,8-bis}{{2-{(2-hydroxy-
`h after drug administration. Thus, despite evidence of inherent
`FeoeeepharinolOst Camnrecanedione, Cancer Res., 40: 1516-
`1518, 1
`is
`tumor sensitivity to mitoxantrone, clinical response obviously
`22. Yap, H-Y., Blumenschein, G. R., Schell, F. C., Buzdar, A., Valdivieso, M., and
`Bodey, G. P. Dihydroxyanthracenedione: a promising new drug in the treat-
`ment of metastatic breast cancer. Ann. Intern. Med., 95: 694-697, 1981.
`
`*D. Woodward (American Cyanamid Co., Pearl River, NY), personal communi-
`cation.
`
`CANCER RESEARCH VOL. 45 APRIL 1985
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