Note : Les descriptions sont présentées dans la langue officielle dans laquelle elles ont été soumises.
~9~354
BACKGROUND OF TH~ INVENTION
__ _ _ _
1. Field of the Invention.
.
This invention relates to stabilized aqueous
solutions of a chemical (cisplatin) used by injection in
the chemotherapy of cancer.
2. Description of the Prior Art.
The platinum compounds are a unique group of
compounds in the antineoplastic group of agents. They
were ~irst noted to have an antibiotic e~fect by Rosenberg
and his colleagues in 1965 and have since been ~ound to be
potent antitumor agents in animals.l'2
Structurally they represent a co~plex formed by
a central atom o~ platinum and surrounded by various
arrangements o~ chlorine atoms or ammonia groups in elther
a cis or trans planar relationship. Two of the more com-
monly studied platinum compounds are diagrammed belGw:
Cl
~ Cl ~NH3
Cl NH3 Cl
Cis-Platinum (II) Cis-Platinum (IV)
Diamminedichloride Di~mminetetrachloride
As can be seen, the platinum compound, cis-
platinum (II) diamminedichloride, selected for clinical
trials by the Natlonal Cancer Institute has the chloride
and amino groups only in the horizontal plane. The cis form
o~ the diamminedichloride complex has been synthesized accord-
ing to the following reaction:
'
--2--
,
NH4Cl
K2[PtC14] ' 2N ~ ~cis-[Pt(N ~ )2C12] + 2KCl
1. Rosenberg~ B., VanC~mp, L. and Krigas, T.,
Inhibition of cell division in Escherichia coli by elec-
trolysis products from a platinum electrodeG Nature
(London) 205: 698_699, 1965.
2. Rosenberg, B., VanCamp~ L.,Trosko, J.E. and
Mansour, V.H., Platinum compounds: A new class of potent
antltumor agents. Nature (London) 222: 385-386, 1969.
3~ Kauf~man, G.B. in J. Kleinberg (Ed.),
Inorganic Synthesis, McGraw-Hill Book Co., Inc., New York,
196~.
The National Cancer Institute has been conduct-
ing clinical tr}als in cancer chemotherapy of the cnemical
for which the United States Adopted Name (USAN) 1s now
cisplatin. Certain information regarding its chemistry and
its pharmaceutical formulation are given in the publication
titled Cllnical Brochure, CIS-PLATINUM (II) DI~MMINEDICHLORIDE
(NSC-119875), H~ Handelman et al., Investiga-tional Drug Branch,
Cancer Iherapy Evaluation Program, Division of Cancer Treat-
ment~ Natlonal Cancer Institute (Revised, August, 1974), on
pages 1~5 and 31-32. The last two pages of Handelman et al.
concern the ~ormulation of cisplatln supplied gratis by the
N C I to clinicians for their clinical evaluation in the
chemotherapy of cancer and read as follows:
s~
PHARMACEUTICAL DATA SHEET
NSC-119875 Cis-Diamminedichloroplatinum (II)
Dosage Formulation
10 mg./vial : The contents of each 20 ml. flint vial
appears as an off~white lyophilized
cake. Each vial contains 10 mg. of
NSC-119875; 90 mg. of Sodium Chloride;
100 mg. of Mannitol and H~drochloric
acid for pH adjustment.
Solution Preparation
10 mg./vial : When reconstltuted with 10 ml. of Sterile
Water ~or Injection, USP3 each ml. of the
resulting solution will contain 1 mg. of
NSC~119875, 10 mg. of Mannitol, and 9 mg.
of Sodium Chloride having a pH range of
3.5-4-5-
Stora~e : The dry~ unopened vials should be stored
at refrigeration temperatures (4-8 C.).
Stability : Intact vials have a provisional stability
of one year when stored at refrigeration
temperature (4-8 C.). Stability recom-
mendations may be adjusted pending com-
pletion of a two-year shelf life study.
Reconstitution as recommended results in
a pale, yellow solution which is stable
for not more than one hour a-t room tem-
perature (22 C.) when exposed to normal
room illuminatlon and not more than eight
hours at room temperature (22 C ) when
protected from light. Reconstituted so-
s~
lutions may form a precipitate after
one hour at re~rigeration temperature
(4 8 C )
Caution : me lyophilized dosage formulations
contain no preservatives and therefore
it is advised to discard solutions
eight hours after reconstitution.
August, 1974
Clinical Drug Distribution Section
Drug Development Branch
Mention of this formulation and additional infor-
mation on its clinical use is given, for example, in Cancer
Chemotherapy Reports, Part 1, Vol. 57, No. 4, pages 465-471
(197~)-
Cancer 30: 1451-1456 (1972) in reference to
cisplatin states that
"The drug material used in this study was
manu~actured by Ben Venue Laboratories Inc.,
Bedford, Ohio. It was supplied by the Cancer
Therapy Evaluation Branch of the National Cancer
Institute in vials containing 10 mg. of cis-
diamminedichloroplatinum, 10 mg. (sic) o~ mannitol
and 9 mg. (sic) of NaCl. The resulting yellowish
white powder dissolved readily ln 8-10 ml. o~
sterile water and was injected immediately
after preparation."
Annals of Internal Med. 86: 803-812 (1977) re-
fers to cisplatin as "DDP" and states that
" m e drug DDP is presently available as
an investigational drug only to qualified
specialists through the Investigational Drug
--5--
3~i~
Branch of the Cancer m erapy Evaluation Programg
National Cancer Institute. The product is
supplied as a white lyophylized powder in vials
containing 10 mg. of DDP, 90 mg. o~ sodium
chloride, 100 mg. of mannitol (U.S.P.), and
hydrochloric acid ~or pH adjustment. When re-
constituted with 10 ml. o~ sterile water for in-
aection (U.S.P.), each ml. of the resulting solu-
tion will contain 1 mg. o~ DDP, 10 mg. of manni-
tol, and 9 mg. of NaCl. The pH of the resulting
solution will be 3.4 to 4.5. At 22 C., the recon-
stltuted solution is stable for at least 8 h."
Thus the formulations described above are sta-ted
to require re~rigeration (4-8 C.) while in vials in the
solid state (i.e.~ before reconstitution), they are difficult
to reconstitute and they have a useful life of only about
twenty hours at room te~perature (22 C.) following reconsti-
tution. The verg act o~ reconstitution can cause problems if
i~properly performed and is better avoided. In addition, be-
cause the aqueous solubility of cisplatin is only about
1 mgm./ml., the cost of preparing dosage forms containing
more than 25 mgm./vial by lyophilization becomes prohibitive.
It was the obâect o~ the present invention to pro-
vide a stable~ therapeutically acceptable, intravenously
in~ectable dosage form o~ cisplatin which would not require
lyophilization and reconstitution, which would not require
re-~rigeration during shipment and storage, and which could
be supplied in dosages o~ 50 mg. or larger.
These obaectives were achieved by the present in-
vention as described in detail below.
5~
SUMMARY OF THE INVENTICN
There is provided by the present invention a stable,
sterile aqueous solution of cisplatin in a sealed container such
as an ampul or ~ial in unit dosage ~orm suitable ~or intra-
venous administration to man, said solutlon having a con- -
centration of cisplatin between about 0.1 and about 1.0
mgm~/ml. and preferably of about 1.0 mgm./ml. and a pH in
the range of 2.0 to 3.0 and preferably in the range of 2.3
to 2.7 and most preferably a pH of about 2.5, said pH
being caused by the presence of the appropriate amount of
a nontoxic~ pharmaceutically and therapeutically accept-
able acid, said acid pre~erably being a strong mineral
acid and most preferably being hydrochloric acid; said
solution optionally containing in addition a nontoxic,
pharmaceutically acceptable, inorganic source o~ chloride
ions in a concentration equi~alent to that produced by the
presence of sodium chloride in a concentration in -the range
of 1 to 20 mgm./m-. and most pre~erably about 9 mgm./ml.;
said solution optionally also being either ~ree of any
other added chemicals or also containing a customary, harm-
less, physiologically acceptable excipient, which is prefer-
ably mannitol, in a concentratlon in the range o~ 2 to
150 mgm./ml. and preferably a concentration o~ about 10
mgm./ml., said solution exhibiting less than 10~ (and
usually less than 4~) loss of potency as measured by high
per~ormance liquid chromatography (HPIC) upon storage ~or
one month at 56 C. Preservati~es can be added i~ desired.
There is further provided by the present inventlon
a method ~or inhibiting the growth o~ a malignant tumor
susceptible to cisplatin which comprises intravenously ad-
ministering to a human af~licted with such a malignant
tumor an amount ef~ective to inhibit the growth of said tumor o~
~ 5~
a stable, sterile aqueous solution of cisplatin in a sealed
container such as an ampul or vial in unit dosage form suitable
for intravenous admlnistration to man~ said solution havin~ a
concentration of cisplatin between about 0.1 and about 1.0
mgm./ml. and preferably of about 1.0 mgm./ml. and a pE in
the range of 2.0 to ~.O and preferably in the range of 2.3
to 2.7 2nd most preferably a pE o~ about 2.5, said pH
being caused by the presence of the appropriate amount of
a nontoxlc~ pharmaceutically and therapeutically accept-
able acid~ ~id acid pre~erably being a ~trong mineral
acid and most preferably being hydrochloric acid; said
solution optionally containing in addition a nontoxic,
pharmaceutically ~cceptable, inorganic source of chloride
ions in a concentration equivalent to that produced by the
presence of sodium chloride in a concentration in the range
of 1 to 20 mgm./ml. and most preferably about 9 mgm./ml.,
said solution option~lly also being either free of any
other added chemicals or also contalning a customa:ry, harm-
less, physiologically acceptable excipient~ which is prefer-
ably mannitol, in a concentration in the range of 2 to
150 mgm./ml. and preferably a concentration of about 10
mgm./ml.~ said solution exhibiting less th~n 10~ (and
usually less than 4%) loss of potency as measured by high
perform~lce liquid chromatography (HPLC) upon storage for
one month at 56 C.
_8--
~995~
DESCRIPTION OF THE PREFERRED EMBODIMENTS
EXAMPLE 1
~ For~ula
: Per Ml. Per Liter
~ Cis-diamminedichloro- A A
: platinum (II) . . ~ . . . 0.0010 g. l.OOO g.
; Sodium Chloride, U.S.P... . 0.0090 g. 9~000 g.
Mannitol, U.S.P. ~ . . . . 0.0100 g. lO.OOO g.
Hydrochloric Acid~
Conc. U.S.P.. . . . . . ~ q.s. to pH q.s. to pH
2.0-3.0 2 0-3 0B
Water ~or Injection,
U.S.P.. . . . . . . . . . q.s. l.O Ml. q.s. lOOO.O Ml.
NOTES:
A. lOO~ basis, adjust weight based on reported purity to
provlde l.O g. lOO~ Cis-diammlnedichloroplatinum (II)
per liter
B. Approximately 0.035 0.050 ml. of 37~ hydrochloric acid
required per gram of sodium chloride to obtaln a p~ of
approximately 2.5.
PRECAUTlONS
Cis-diamminedichloroplatinum (II) is a toxic sub-
stance and is listed on page 942 in the 1976 edition of the
"Registr~ of Toxlc E~ects o-f Chemical Substances~" The
OSHA Standard of Time Weighted A~erage (TWA) ls 2 mcg./m3.
_g _
Consult the above, listed references, pertinent local pub-
lications and regulations and such publications as the
National Cancer Institute Safety Standards for Research
Involving Chemical Carcinogens and the National Institute
of Health Specifications for a Class II type 1 Safety
Cabinet. Any cis-dia~minedichloroplatinum (II) weighing,
the working surface of the batching vessel, the filling,
stoppering9 and sealing must be provided with such protec-
tion.
All personnel invol~ed with compounding of this
product must be protected with full nylon head/face cover,
coveralls, rubber gloves and a respirator equivalent to the
MSA Ultra Filter Respirator rated for environments contaml-
nated with dusts, fumes and mists having a TWA rating of
less than 50 mcg./m~ During the sterile liquid filling
the sterile head/face cover, surgeon's gauze mask and goggles
can replace the respirator. Any uniforms grossly contami-
nated due to spills, etc. should be stored in a closed meta
container until burned.
THE IMPORTANCE OF PROTECTING PERSONNEL DURING T~E
HANDLlNG, MANUFACTURING AND ASSAYING OF THIS PRODUCT IN
_ _ _ . _ _ _ _ _ _
ACCORDANCE WITH THE ABOVE CANNOT BE O-~EREMPHASIZED.
_
The prime bulk cis-dia~minedichloroplatinum (II)
must be protected from light. The processing and filling of
vials described herein was conducted under diffused
natural/fluorescent light.
Batching ~essel
Glass-lined, agitated, pressure ~essel A ~16 SS
agitator is permissible. Working volume must be consistent
with batch size. A dipstick and calibration cur~e of tank
--10--
J~
~ 5 ~
volume is requlred for determining volume.
Milli~ore Mem~rane Filter Holders
316 SS. Fllter area with necessary pre-filters
and 0.22 micron final sterilizing f~lter.
Transf
* *
Teflon or Tygon.
All stainless s-teel contacts should meet 316 SS
requirements. All other equipment should be appropriate to
produce a sterlle, non-pyrogenic, particulate-free product.
Manufacturing Instructions
A. m ese instructions are written for an eight-
hour batchin~ to filling operation. Storage o~ this product
before filling has not been in~estigated at this writing.
B. Maintain 27 C. ~ 2 C. temperature conditions
throughout entire batching and filtering operations.
1. Place 80~ of the batch volume of Water for Injec-
tion, U.S.P. in a suitable vessel.
2. With agita-tion add the sodium chloride. Agitate
ten minutes or until dissolved.
~ . With good agitation carefully ad~ust the pH of the
sodium chloride solution to 2.0-3.0 (pre~erably 2.5) with
concentrated hydrochloric acid. See estimated amount under
note "B" on formula sheet. Agitate ~or ten minutes a~ter
last addition. Recheck pH~
4. With good agitation add the mannitol and agita-te
ten mlnwtes or untll dissolved.
5~ With good agitation and tak~ng special precautions
against dusting and exposure, add the cis-diamminedichloro-
*Trade Marks
platinum (II). Rinse its container sufficiently wi-th an
appropriate amount of water for injection and add to the
batch.
6. Agitate until completely dissolved. Approximately
60-go minutes will be required for co~plete dissolution.
Monitor pH and add additional concentrated hydrochloric acid
if required to maintain at 2.0-3.0 (optim~n 2.5).
7. Carefully adjust volume to theoretical batch ~olume
wlth water for injection. Make final pH check.
8. PasS the solution through a clean, sterile 0.22
micron Millipore Filter into the sterile filling line.
9. Fill as directed below for the following products:
10 Mg.~Vial
Sterile, Type I ~mber, 15 ml. vial, with a 10-ml. fill.
Stopper with red, 20mm Teflon-~aced stoppers and seal with
aluminum seals. Numbered as K93, 100 and
107 with nitrogen overlay and K94, 101 and 108 without
nitrogen overlay.
2~ ~ ial
Sterile, Type I amber, 50 ml~ vial with a 25-ml. fill.
Stopper with red, 20mm Teflon faced stoppers and seal with
aluminum seals. Numbered as K95, 102 and 109
with nitrogen overlay and K96, 103 and 110 without nitrogen
overlay.
50 M~./Vial
5terile, Type I amber~ 50 ml. vial with a 50-ml. fill.
Stopper with red, 20mm Teflon-faced stoppers and seal with
aluminum seals Numbered as K97, 104 and 111
with nitrogen overlay and K9~, 105 and 112 without nitrogen
overlay.
54
I~ese formulations were prepared in ~ive groups
(with the final pH given ln parentheses) as follows:
K9~-96 (pH 2~4)
K97-98 (pH 2.5)
K100-103 (pH 2.3)
K104-105 (p~ 2.4)
K107~110 (pH 2.~)
K111-112 (p~ 2.4)
Original potencies by HPLC assay were in the range o~ 0.99
to 1.00 mgm./ml.
m e percentage loss in potency after storage ~or
one or two months at the indicated -temperature was ~ound by
; HPIC assay to be as follows:
: 56 C. - 45o C
One Two One
Month Months Month
K93 -~F~5~
K94 2.0 4.0
K95 o,o -1.0*
K96 1.0 4.0 0.0
K97 .3
K98 4.0 2.0
K100 3.0
K101 ~.0 5.0
K102 3.0
K103 5.0 4-
K104 0,0 -I.O*
K105 0.0 5~
K107 1.0 0.0
K108 2.0 5.0 1.0
K109 3.0
KllO 3.0
Klll 7.0
K112 7.0
*Negative sign means assay showed 1.0~ increase in potenc~.
,.
~995~
The above-described solutions, with and without
nitrogen cover, thus have shown 7~ or less loss in potency
after storage at 56 C. and 45 C. with the majority show-
ing a loss of potency of ~ or less. The pH of the solu-
tions remained between 2.4 and 2.7.
Physically, no change is apparent at 56 C. or
45 C. after one month. Solutions remain clear and
colorless. Initial Klett readings averaged 8-12, one-
month 56 C. and 45 C. readings averaged 6-15 No
changes or differences are noted between samples with or
without N ~.
One sample at each temperature station for all
products was tested inverted exposing the solution to the
Teflon-coated plug stopper. Samples from inverted products
were assayed from 56 C. at one month with and without N2
exposure. Stability was not affected at one month 56 C.
as samples showed only 1-2~ loss of potency.
At two weeks 4 C. samples were observed for
crystallization of cis-di~mminedichloroplatinum (II). No
crystals were obser~ed until one month and it was only
noted randomly, not in every sample. Only one lot o~ the
10 mg~/~ial and 25 mg./vial products show some crystals
forming rand~mly at 4 ~. Crystallization is noted through-
out all lots of 50 mg./vial products but again not in all
samples One sample from 4 C. with crystals could not be
redissolved by warming the solu-tion to 37 C. with agita-
tion. Only partial success was obtained. It appears these
products cannot be stored under refrigerated cond.itions as
even redissolving of crystallized products was difficult.
Cis-platinum (II) diamminedichloride (NSC 11987~)
is an inorganic compound ~irst noted to prevent replication
of E. coli and subsequently ~ound to possess antitumor
_.
activity. The drug exerts its effect of interfering with
DNA synthesis by causlng cross-linking of complementary
strands o~ DNA. It has activity in a variety of tumor sys-
tems including L1210, Sarcoma 180, Walker 256 carcinosarcoma~
DMBA induced mammary tumors and ascitic B16 melanosarcoma.
The compound is especially interesting in that i-t exhibits
synergism with a large number of currently-used chemothera-
peutic agents Large animal toxicology studies showed renal
tubular necrosis, enterocolitis~ bone marrow hypoplasia and
lymphoid atrophy. Phase I studies have demonstrated the fol-
lowîng toxicities: myelos~ppression, renal insufficiency,
high frequency ototo~icity and GI intolerance. Currently
u~ed dosages with mild to moderately acceptable toxicity are
in the range o~ 60-100 mg/m2 IV as à single dose or divided
over 3-5 days, to be repeated at four-week intervals. Early
clinical trials show some responses to the drug in germinal
cell tumors, lymphomas, sarcomas, breast and head and neck
carcinomas.
A dosage of 60 mgm/m2 is roughly equal to 1.5 mgm/kg
which in turn is roughly equal to 105 mgm/patient weighing
70 kg.
The solutlons of the present invention are used in
the same manner and for the same purpose as stated above and
in the other publications and in the volumlnous medical
literature on this sub~ect. As stated therein, frequent use
is made of concurrent therapy with other chemotherapeutic
agents for best results. ~hen desired~ the solu-tions of -the
prese~t invention may be added immediately before use to a
-15-
95~
sterile, pharmaceutically acceptable aqueous diluent such
as glucose or saline. Administration is either by direct
intravenous injection or by intravenous infusion.
:
-16-
- ' '
35~L
cis-Diamminedichloro~latinum.
Method
Cis-diammined~chloroplatin~ is chromatographed
on a Water's ~-Bondapak-NH2 column using a loop inaection
technique. Detection is achieved by monitoring the U.V.
absorbance at 313 r~ and quantitation is acco~plished by
peak height measurement with external calibration, This
method is applicable to bulk powders and solid dose for-
mulations containing NaCl and mannitol. Specificity has
been demonstrated by separation o~ the cis and trans isomers,
and apparent degradations (moisture~ acid, base, heat and
accelerated light).
NH7 Cl Cl NH
/ \ / 3
Pt Pt
/\' /\
NH3 Cl 3 Cl
Cis-(NH3)2 C12 Pt II ( 3)2 2
r,~ rD-
Column - Water's Micro Bondapak-NH2 (300 MM X 4.0 MM ID)
027386 or equivalent.
obile Phase - Ethyl acetate/methanol/dimethyl~ormamide/
distilled water (25/16/5/5). Use Burdick
and Jackson distilled in glass spectro-
quality reagents. ~egas the wa-ter prior
to use and the solution after mixing.
etector - Water's Model 440 Absorbance Detec~orO
avelength - 313 ~m (U.V.).
-17-
~ 5
Sensitivity - O.1 AUFS.
In~ector - A 20 microliter loop injector.
The Loop Injector - A Valco 7000 psi stainless steel
valve (CV -6-UHPa-C20).
Injection Volume - 20 mlcroliter.
Solvent Delivery System - Water's Model 6000A pump.
Flow 2.0 ml~/minute,
Retention - 2.8 minutes (appro~.).
Recorder Heath Model SR-2~B.
Chart Speed - O.5 inch~mln.
Range - 10 milli~olt.
Using the conditions above, obtain chromatograms
o~ the standard and sample preparations in duplicate.
Re~erence Standard of cis diamminedichloro-
platinum (DDP):
Lot No, = 78F7 (Matthey Bishop Lot No. AM7702)
Asslgned Purity = 99.8~
501vents - Burdick and Jackson (distilled in glass) spec-
troquality.
Standard - Weigh accurately 25 mg. of cis-diamminedichloro~
platinum (DDP) into a 25-ml. ~olumetric ~lask.
Dissolve in and dilute to volume with dimethyl-
~ormamide.
L~ophll~zed In~ection - Reconstitute vial contents with
10.0 ml. o~ dimethylformamide ~ld mechanically
shake ~or 5 minutes (alternately a sonic bath
may b~ used for 2 minutes). Filter 5.0 ml. of
the sample solution (Millipore Filter Kit or
*Trade Mark
-18-
~1
95~
equivalent) discarding the first ml.
ontent Uniformity - Prepare 10 vials as described above
and assay,
Calculations
__
STANDARD FACTOR (SF) = AV~RAGE PEAK HEIGHT ST~NDARD
MG DDP/GRAM = SF X AVERAGE PEAK ~EIGHT SAMPLE X 25
MG. DDP/VI~L = SF X AVERAGE PEAK HEIGHT SAMPLE X 10
For assay, average results obtained for 10 vials
from content uniformity test.
Me
The method of assay for cisplatin described above
is applied to aqueous solutions (dose formulations) con
taining NaC1 and mannitol after making the changes indicated
below.
~PLC Conditions
_ _ _obile Pha~e - Acetonitrile/distilled water (75/25, v/v).
Use Burdick and Jackson distilled in gla~s spec-
troquality reagents. Degas the water prior to
use and the solution after mixing.
Injector ~ A100 microliter loop injector.
Injection Volume - 100 microliter.
Retention - 2.0 minutes (approx.).
H ~
Sta~dard - Weigh accurately 25 mg. of cis-diamminedichloro-
platinum (DDP)~ 225 mg. sodium chloride and 250 mg.
-19-
S4
mannitol into a 25-ml. volumetric flask. Dissolve
in and dilute to vol~me with distilled water,
Pipet 5 . O ml, of the resulting solution into a
25,-ml. volumetric flask, add 2.0 ml, of dis-
tilled water and take to volume with ac etonitrile .
Sample (1 mg./ml.) Pipet 5.0 ml. of the sample into a
25 ml. volumetric flaskg add 2.0 ml. of d~stilled
water and take to ~olume with acetonitrile.
Calculations
____ ,
STANDARD FACTOR (SF) =
MG, DDP/ML. = SF X AVERAGE PEAK HEIGHT SAMPIE X 25
This invention is capable of industrial ~pplication.
c.O~
