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pound that was designed to be delivered in a multilamellar liposome by substituting the chloride
for neodecanoato leaving groups, which results
in an enhanced affinity for biological membranes.
Free NDDP is devoid of in-vivo antitumor activity. 11 In the current L-NDDP formulation, the liposome is composed of dimyristoylphosphatidyl
choline (DMPC) and dimyristoylphosphatidyl
glycerol (DMPG) in a 7:3 molar ratio and the
lipid-to-NDDP ratio is 15:1, whose acidic phospholipid DMPG component is essential for antitumor activity. 12 The resulting multilamellar liposome measures 13 m m in diameter and acts
as a vehicle and slow-release system for the drug.
The organ distribution of platinum species is
typically determined by first collecting fluids or
tissues and then subjecting them to atomic absorption (AAS) for elemental analysis or to liquid scintillation counting (LSC). Although other
techniques are available, such as X-ray fluorescence (XRF) and inductively coupled plasma
mass spectrometry (ICP-MS), their use is limited
by, either insufficient detection limits (in the case
of XRF), or expense and labor (in the case of
ICP-MS). Neutron activation analysis (NAA) has
the best detection limits among assays for platinum. The comparative platinum limits for the
various methodologies are 0.08 m g/mL for ICPMS, 20 m g/mL for inductively coupled plasmaoptical emission spectrometry (ICP-OES), 4
m g/mL for graphite furnace AAS (GFAAS), 100
m g/mL for flame AAS (FAAS) 13 , 0.005 m g/mL
for NAA, and 50000 m g/mL for XRF.
According to the literature, most assays for
platinum radioisotopes start with isotopically
pure Pt-195m or with Pt-191. The enriched form
of Pt is then processed into the synthesized compound of interest. Akaboshi et al.14 used enriched
Pt-195m-radiolabelled trans and cis forms of
diaminodichlorop latinum (II) to compare the
killing efficiency of the two isomers in HeLa
cells. Hoeschele et al.15 intravenously injected
enriched Pt-195m -labeled cis and trans dichlorodiammine platinum (II) to study the tissue distribution. Afterward, the organs were harvested and
analyzed by LSC. LSC was also used by Desimone et al.16 to determine the distribution of Pt195m-labeled cisplatin. Bnard et al.17 also used
Pt-195m-labeled cisplatin, but opted for autoradiography to determine the tissue distribution.
Robins et al.18 injected accelerator produced Pt191 ethylenediamine dichloride platinum (II) into
rats and assessed its activity using LSC.
In this work, we used NAA to activate the plat254
Figure 1.
RESULTS
DISCUSSION
The objective of our study was to determine the
feasibility of radioisotopically activating the platinum-containing compound NDDP, a DACHplatinum compound currently undergoing clinical studies as part of a liposomal formulation.
All currently used platinum anticancer agents
undergo intracellular activation by acidic hydrolysis, loss of the leaving groups, and formation of
aquated species, which are thought to be the ac-
Elemental Analysis
Upon analysis by HPGE, various radioisotopes
of platinum could be seen, along with potassium
and sodium . Figure 3 shows the spectrum of activated NDDP counted for 1 hour on an HPGE
detector after a 3 hour irradiation and a 5 day decay period. Silver and iodine may have been present, but were not seen at 5 days because either
they were minor constituents or they decayed
away prior to analysis at 5 days.
Stability of NDDP Powder
The heat of the reactor core caused a partial
breakdown of NDDP, as evidenced by a color
change of NDDP from white to tan. The initial
HPLC data appeared to indicate that 65% of the
NDDP had degraded away during irradiation,
shipping, and decay prior to the analysis (NDDP
elutes at 8.59.0 minutes). Fractions collected
and analyzed by AAS gave only 33% of the platinum signal expected, indicating that water may
have worked its way into the supposedly sealed
vials during irradiation.
255
Figure 3. Spectrum of 4 mg of activated NDDP counted for 1 hour on an HPGE detector after 3 hours of irradiation and a 5day decay period. (Top) 50-225 keV region indicating the abundant emission of X-rays by iridium, platinum , gold, and mercury.
(Bottom) 50-600 keV region, indicating the abundant emission of photons by platinum -191.
Half-life
Time 0
24 hours
5 days
10 days
Actual
activity at
5 days
2.96 days
4.33 days
50 years
4.02 days
95.4 min
18.3 hours
13.5 sec
30.8 min
4.93 sec
10.6 days
3.2 days
42.6 min
2910
198
0.474
886
11700
25700
27300
339000
2910
0.474
0
0
2310
168
0.474
745
0.333
10400
0
0
2310
0.474
1870
1870
904
88.8
0.474
374
0
273
0
0
904
0.474
722
722
280
39.9
0.474
158
0
2.9
0
0
280
0.474
256
256
8.88
356
37.4
139
109.5
184
204
Anticipated Activity
Isotope
191-Pt
193m-Pt
193-Pt
195m-Pt
197m-Pt
197-Pt
199m-Pt
199-Pt
191m-Ir
193m-Ir
199-Au
199m-Hg
Isotopes
seen
Energy
(keV)
Decayed
isotopes
Na-24
K-40
1368.6
1460
Pt-191
82.4
96.3
129.4
172.2
351.2
368.7
409
456.5
539
624.1
135.5
98.9
129.8
191.4
129.4
158.4
208.2
129.4
Pt-197
Pt-197m
Pt-19m
Pt-199m
Pt-193m
Pt-195m
Pt-197
Ir-191m
Au-199
Hg-199m
resistance to platinum compounds may be secondary to reduced or slow intracellular drug activation.
Currently, abundant natural isotopic platinum
Isotopes with
low potential
Pt-193
K X-rays of Pt, Ir, Hg at 60 to 80 keV also seen from the decay of the numerous platinum
species.
For Na-24 the 2754 keV was not seen; upper threshold of detector set at 2000 keV
257
4. Perez-Soler R, Khokhar AR, Lopez-Bereste in G. Treatment and prophylaxis of experimental liver metastases
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5. Perez-Soler R, Lopez-B erestein G, Lauterszt ain J, et
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improved stability and preserved antitumor activity with
complexes containing linear alkyl carboxylato leaving
groups. Cancer Chemother Pharmacol 1994; 33: 378.
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Khokhar AR. Increased cytotoxicity and reversal of resistance to cis-diammined ichloroplatinum (II) with entrapment of cis-bis-neodecanoa to-trans-R,R-1 ,2-diamminocyclohexane platinum (II) in multilam ellar lipid
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K, Miyahara T. Determination of the target volume of
HeLa cells treated with platinum -195m radiolabeled
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259