United States Patent [19J
`Cleare et al.
`
`[11]
`
`[45]
`
`4,140,707
`Feb. 20, 1979
`
`[75]
`
`[54] MALONATO PLATINUM ANTI-TUMOR
`COMPOUNDS
`Inventors: Michael J. Oeare; James D.
`Hoeschele; Barnett Rosenberg;
`Loretta L. Van Camp, all of East
`Lansing, Mich.
`[73] Assignee: Research Corporation, New Yotk,
`N.Y.
`[21] Appl. No.: 778,955
`Mar. 18, 1977
`[22] Filed:
`
`Related U.S. Application Data
`[63) Continuation of Ser. No. 260,989, Jun. 8, 1972,
`abandoned.
`Int. 0.2 .............................................. C07F 15/00
`[51]
`[52] u.s. a ................................. 260/429 R; 424/245;
`424/287; 546/4
`[58] Field of Search .................................... 260/429 R
`
`[56]
`
`References Cited
`PUBLICATIONS
`Ward et al., Cancer Treatment Reports, vol. 60, No. 11,
`1675-1678 (1976).
`Rosenberg Plat. Metal Rev. 15, pp. 42-47 (1971).
`Leh et al., J. Pharmaceutical Sciences, 65, No.3 (1976),
`pp. 315-320.
`Rosenberg et al., Nature 222, 385-386 (1969).
`
`Primary Examiner-Helen M. S. Sneed
`Attorney, Agent, or Firm-Dennis P. Clarke
`
`ABSTRACT
`[57]
`Malonato platinum coordination compounds and a
`method of treating malignant tumors comprising the
`parenteral administration to an affected animal of a
`solution of the compound.
`
`4 Oaims, No Drawings
`
`NEPTUNE GENERICS 1006- 00001
`
`APOTEX 1006-0001
`
`

`
`1
`
`4,140,707
`
`MALONATO PLATINUM ANTI-TUMOR
`COMPOUNDS
`
`The invention described herein was made in the
`course of work under a grant or award from the De(cid:173)
`partment of Health, Education and Welfare.
`This is a continuation, of application Ser. No.
`260,989, filed June 8, 1972, now abandoned.
`
`BACKGROUND OF THE INVENTION
`The present invention relates to novel malonate plati(cid:173)
`num coordination compounds and to their use in cancer
`chemotherapy.
`
`SUMMARY OF THE INVENTION
`The invention provides platinum coordination com(cid:173)
`pounds having the formula:
`
`2
`num (IV) forms d2sp3 coordination compounds which
`have an octahedral arrangement in space.
`The coordination compounds of the invention in(cid:173)
`clude the cis and trans isomers of platinum (II) and
`5 platinum (IV) which contain the bidentate malonate
`ligand which may be substituted or unsubstituted. The
`malonate ligand may contain substituents selected from
`the group consisting oflower alkyl, (e.g., methyl, ethyl,
`n-propyl, isopropyl, n-butyl, etc.); aryl, (e.g., phenyl;
`10 lower alkyl-, lower alkenyl-, halo-, nitro-, lower alkoxy(cid:173)
`substituted phenyl and naphthyl); aralkyl, (e.g., phenyl(cid:173)
`methyl (benzyl), 2-(1-naphthyl)methyl); alkenyl, (e.g.,
`4-amino-1-butene, allyl); cycloalkyl, (e.g., cyclopropyl,
`cyclohexyl, etc.); cycloalkenyl, (e.g., 2-cyclopenten-
`15 1-yl, 2-cyclohexen-1-yl); alkoxy, (e.g., methoxy, ethoxy,
`etc.), and hydroxy. Also suitable are the ·1,1-cycloalk(cid:173)
`ylenedicarboxylic
`acids,
`(e.g.,
`1,1-cyclo(cid:173)
`propanedicarobxylic acid, 1, 1-cyclobutanedicarboxylic
`acid, etc.) and the 1,1-cycloalkenyldicarboxylic acids,
`20 (e.g.,
`1,1-cyclopropenedicarboxylic
`acid,
`l, 1-
`cyclobutenedicarboxylic acid, etc.)
`The coordination compounds of the invention also
`contain two monodentate ammonia or primary or heter-
`ocyclic amine ligands, i.e., when x in the above formula
`wherein:
`x = 1 or 2;
`25 is 2 or one bidenta:te amine ligand, i.e., when x is l.
`y = 1 or 2:
`Suitable monodentate amine ligands include lower
`z = o, 1 or 2,
`alkyl amines, (e.g., methyl-, ethyl-, n-propyl-, isopropyl-
`, n-butyl- amines, etc.), aryl amines, (e.g., aniline), aral-
`provided that when y = 2, z = 0 and when y = 1, z is
`greater than O;
`kyl amines, (e.g., benzylamine), hydroxy lower alkyl
`R and R 1 are selected from the group consisting of H, 30 amines, (e.g., ethanolamine, propano1amine; etc), by-
`lower alkyl, aryl, aralkyl, alkeny1, cycloalkyl, cy-
`droxylamine, lower alkoxy amines (e.g., methoxyla-
`cloalkenyl, alkoxy, OH, or are combined with the car-
`mine, etc.), alkoxyalkylamines (e.g., methoxymethyla-
`bon atom to form a cycloalkyl or cycloalkenyl group,
`mine, etc.), and heterocyclic amines (e.g., pyridine and
`and substituted derivatives thereof;
`aziridine). Also included are the amino acids, i.e.,
`when x = 1, A is HR2N-CHR3-CH~-NRsH 35 R7-CHNH2-COOH wherein R7 is H, lower alkyl
`and when x = 2, A is H2NR6a heterocyclic amine or an
`(e.g., methyl, isopropyl, etc.), hydroxy lower alkyl (e.g.,
`hydroxymethyl, hydroxyethyl, etc.), aralkyl (e.g., ben-
`amino acid, wherein R2, RJ, R4 and Rs are the same or
`zy~t ei~c)~o be understood that the coordination com-
`different and are selected from the group consisting of
`H, CH3, C2Hs, hydroxy and lower alkoxy provided that 40 pounds of the invention may include two identical or
`different monodentate ligands.
`R2 and Rs may also be aryl or aralkyl, and each R6 is the
`same or different and is selected from the group consist-
`Suitable bidentate amine ligands include the substi-
`ing of H, lower alkyl, aryl, aralkyl, hydroxy lower
`tuted and unsubstituted primary and secondary
`alkyl, hydroxyl and alkoxyl amines, alkoxylalkylamines
`ethylenediamines. One or both of the carbon atoms of
`wherein all of said alkyl groups are lower alkyls and 45 the ethylenediamine may contain substituents such as
`heterocyclic substituents including said N as a ring
`lower alkyl (e.g., methyl, ethyl), hydroxyl, alkoxy (e.g.,
`member;
`methoxy, ethoxy, etc). Secondary ethylenediamines
`when z = 1, L ~s a bidentate anioni~ li~~d, and
`wherein one or more of the amine groups contains sub-
`when z = 2, L IS a monodentate antomc hgand.
`stituents such as listed above for the carbon atoms ofthe
`The inventio~ also ~elates to a c?m~sition and so primary amine and aryl (e.g., phenyl) and aralkyl(, e.g.
`benzyl) may also be utilized.
`m:t~od for treatmg mal~~ant. tumors m ~mals com-
`p~smg pa~enterally adm1mster:tng to an. a?1mal aff:cted
`The Pt (II) coordination compounds specified herein
`w1th a. ma~1gnant tumor a solution contam!ng a plat!num
`do not exist as geometrical isomers; however, the Pt
`coordmation 7ompound as define_d heremabove m an
`(IV) compounds exist as cis and trans isomers. It is to be
`amount sufficient to cause regression of the tumor.
`55 further understood that the invention is inclusive of the
`cis and trans isomers.
`DETAILED DESCRIPTION OF THE
`The Pt (IV) coordination compounds may also con(cid:173)
`INVENTION
`tain two monodentate or one bidentate anionic ligand
`Platinum coordination compounds and methods for
`where only one malonato ligand is present, i.e., where y
`= 1 in the above formula.
`their production are described by J. C. Bailar, Jr., The 60
`Chemistry of the Coordination Compounds, Reinhold
`Suitable monodentate anionic ligands include chlo(cid:173)
`Publishing Corp., N.Y., 1956, Chap. 2; J. Lewis et al,
`ride, bromide, iodide, nitrite, hydroxide, nitrate, sulfa(cid:173)
`Modern Coordination Chemistry: Principles and Methods,
`mate, etc. Among the bidentate anionic ligands which
`Interscience Publishers, Inc., N.Y., 1960 and Kauffman
`may be present are oxalate, pyrophosphate, dithioxa-
`Inorganic Synthesis, 7, McGraw-Hill Book Co., Inc.,
`65 late.
`N.Y., 1963.
`It is to be understood that the invention includes
`Platinum (II) forms dsp2 coordination compounds
`those coordination compounds containing mixed mono(cid:173)
`dentate anionic ligands;,
`which have a square planar arrangement in space. Plati-
`
`[Pt(lllAx(OOCh-CRRtl or
`
`cis or trans[Pt(IV)Ax(OOC)z-CRR2)yLz]
`
`NEPTUNE GENERICS 1006-00002
`
`APOTEX 1 006 - 0002
`
`

`
`4,140,707.
`
`4
`3
`(to avoid "bumping") on the hot plate until white crys-
`The preferred compounds are those wherein Rand
`R 1 in the above formula are H, methyl or ethyl, i.e.,
`tals of the product started to form in great quantity. The
`mixture was then cooled to room temperature and the
`malonatoplatinum, methylmalonatoplatinum and ethyl-
`product filtered off. The filtrate was reheated for 5-10
`malonatoplatinum coordination compounds. The most
`preferred Pt (II) compounds are those malonato- 5 minutes and cooled to o• C. to collect a further crop.
`The crude yield at this stage was 20.5g (93% ).
`platinum (II) compounds of the above formula wherein
`x = 1 and R2, R3, ~and Rs are each H, i.e., malonatoe-
`The product was recrystallized by dissolving in boil-
`(II), methylmalonatoe-
`ing or near boiling. water. The above yield (20.5g) re-
`thylenediamine
`platinum
`(II) and
`ethylma1onatoe-
`quired about 3 liters of boiling water for complete disso-
`thylenediamineplatinum
`thylenediamineplatinum (II); and wherein x = 2 and 10 lution. Malonic acid 1g/L was dissolved in the water to
`suppress any hydrolysis. • The fJltered solution was
`each R6 is H, i.e., malonatodiammineplatinum (II), me-
`to o• C.
`and
`ethyl-
`cooled
`to give white fluffy needles
`thylmalonatodiammineplatinum
`(II)
`(18.25g-83%).
`.
`malonatodiammineplatinum (II).
`.
`•u. V./via spectral and conductivity studies have shown that hydrolysis
`The preferred Pt (IV) compounds are those wherem
`x = 2, each R6 is H and y = 2, i.e., bismalonato (or 15 is negligible.
`bismethylmalonato or bisethylmalonato) diammine plat-
`The crystals decompose between 185"-190" C. The
`inurn (IV).
`structure of the product was verified cia an i.r. spec-
`The coordination compounds of the invention may be
`trum. Solubility ofthe product is low in cold water, i.e.,
`prepared by one of a variety of well-known methods. A
`20 mg/100 mls at 20" C. and 43 mg/100 mls at 37' C.,
`general method of prepamtion of the Pt (II) coordina- 20 but higher in near boiling water (90" -1 oo• C.)-65g/100
`tion compounds is as follows: Starting compounds hav-
`ml.
`ing the formula cis-[Pt A(Halh] wherein Hal is I, Cl or
`The empirical composition was verified by elemental
`Br and A is one bidentate or two monodentate amine
`analysis:
`ligands (prepared by the method of S. C. Ohara, Indian
`Malonatodiammineplatinum(II) [Pt(NH3)2(C3H204))
`J. Chern., Vol 8, p. 193 (1970)) are reacted with silver 25
`Calculated for C3H8N20 4Pt.C.: 10.88; H: 2.43; N:
`nitrate to form the diaquo complex. The latter is then
`8.46: Pt 58.9; Found C: 10.67; H: 2.35; N: 8.54; Pt 58.7.
`reacted with the malonate ion to form the coordination
`compounds of the invention. This method is repre-
`sented by the following reaction scheme:
`cis-[Pt ACiu + 2AgN03 + 2H20) __,. cis-[Pt
`A(H20) 2l(N03)2 + 2AgCI
`
`30
`
`EXAMPLE 2
`[Pt (en) (CA04)] (en= H2N(CH2)2NH2; CA042-
`= 02C CH(CH3) C022-)
`Silver nitrate (3.64g) was dissolved in 20 rn1 of water
`and added to [Pt(NH2MCH2hCL2] (3.5g) suspended in
`water (30 ml.) in a conical flask. The mixture was stirred
`on a warm hot plate for 5-lO minutes until all the yel-
`35 low platinum complex had dissolved to give a yellow
`liquor plus a copious white silver chloride precipitate.
`The mixture was flltered through a fine pore r.tter and
`the precipitate washed twice with small volumes of hot
`water. The clear filtrate plus washings was added to an
`40 aqueous solution of methylmalonic acid (2g in 20mls)
`which had been adjusted to pH 5-6. The mixture was
`heated to about so· c. for five minutes and then cooled
`to 0~ C. The shiny white crystals which formed were
`filtered and washed with cold water and acetone (Yield
`45 2.65g). The mother liquor plus aquoous washings was
`reduced to about half its original volume (- 30 rnls) to
`yield a second crop on cooling to o• C. (Yield 0.85g).
`Total Crude yi,eld was 3~50 gms (88%). The complex
`was recrystallized from a minimum volume of boiling
`50 water (around 250 mls) with fdtration through a fine
`pore filter prior to cooling to Q" C.
`Yield of shiny white leaflets 2.96g (74% ).
`. Calculated for C~12N20~: C:l9.41 H 3.26 N:7.55;
`Found C:19.11 H 3.61 N:7.89 ..
`A second crop (0.33g-8%) was obtained by reducing
`the bulk of the mother liquor.
`
`cis-[Pt A(H20)u(N03h + H2C-(C00)2 __, [Pt
`A(OOC)2-CHu + 2N03 - + 2H20
`
`wherein A is one bidentate amine ligand or two mono-
`. .
`.
`.
`dentate amine_ ligands. .
`The followmg non-hmitu?-g examples are Illustrative
`of the methods for preparmg the compounds of the
`invention.
`
`EXAMPLE 1
`Malonatodiammineplatinum(II)
`[Pt(NH 3)2(C3H204)]
`
`Reactions:
`
`[Pt(NHJhCI:z] + 2AgN03 + 2H20) __,.
`[Pt(NH3h(H20)u(N03) + 2AgCI
`
`[Pt(NH3h(H20)u(N03h + C3H2042- __,.
`[Pt(NH3)2(C3H204)] + 2N03- +2H20.
`
`11
`
`slightly less than the stoi(cid:173)
`Silver nitrate (22.55g -
`chiometric amount in order to avoid silver contamina(cid:173)
`tion) was dissolved in water (50 mi.) and added to 55
`[Pt(NH3)2Cl2] (20g) in a 250 mi. conical flask. The con(cid:173)
`tents were warmed (60" C.) on a hot plate with rapid
`stirring until the silver chloride precipitation was com(cid:173)
`plete and the mother liquor was almost colorless. The
`silver chloride was filtered off using a fine pore sintered 60
`glass filter and the precipitate was washed several times
`with hot water to give a total filtrate volume of 100-200
`ml.
`Malonic acid (13g- a twofold excess) was dissolved
`in water (30 mi.) and neutralized with a solution of 65
`KOH ( -13g in 30 mi.) to pH 5-6. The resulting potas(cid:173)
`sium malonate solution was added to the platinum con(cid:173)
`taining filtrate and the mixture was carefully warmed
`
`EXAMPLE3
`traris-[Pt IV(NH3)2(mal)2)
`Silver nitrate (5.45g) was dissolved in water (30 rnl)
`and added to trans [Pt(NH3)2C14] (3g) suspended in
`water (30 mls) containing. concentrated nitric acid (3
`ml), The. contents were warmed on a hot plate (70' -so·
`C.) and stirred for at least one hour. The mixture was
`filtered through a fine pore sintered glass filter to re(cid:173)
`move the silver chloride. The precipitate was washed
`twice with a small volume of hot water. The clear fd-
`
`NEPTUNE GENERICS 1006-00003
`
`APOTEX 1 006 - 0003
`
`

`
`4,140,707
`
`5
`6
`following standard protocols for this testing as set by
`trate plus washings was tested with a drop of 1M KCl
`the National Cancer Institute. (Cancer Chemotherapy
`solutions to determine if excess silver chloride was pres-
`Rep., 25(1962)).
`ent. (If the test is positive, sufficient KCl is added drop-
`wise to the bulk solution until no silver chloride is pre-
`For these tests an S 180 tumor taken from a sacrificed
`cipitated.) The solution was refiltered and the filtrate 5 mouse was disected free of superfluous tissue and cut
`reduced to 20-30 mls in volume and cooled to 0' C. to
`under sterile conditions into approximately 10 milli-
`yield plate yellow
`crystals
`(presumably
`trans
`gram size pieces. These tissue pieces were then im-
`(Pt(NH3~2(N03)4]). These were washed with a little
`planted by trocar in the left axillary region, subcutane-
`cold water and then acetone (Yield 1.8g). A portion of
`ously, in new mice. The mice were, on the average,
`this yield (lg) was dissolved in a minimum of hot water 10 approximately four weeks old and weighed 18-20
`to which sodium nitrate (0.2g) had been added. This
`grams. Taking day 0 as the day of implant, the animals
`solution was filtered into an aqueous solution of malonic
`were sacrificed on day 10. The tumors were excised and
`acid (0.5g- a slight excess) which had been adjusted to
`weighed and the ratio of the weights of the tumors in
`pH 5-6 with sodium hydroxide. White nucro-crystals of mice in the treated animals to the control set of animals
`the complex quickly form on cooling. These were fil- 15 was obtained. This ratio, multiplied by 100, is given as
`the T /C ratio in Table I.
`tered off and washed with cold water and acetone.
`(Yield 0.7g- 30-40%).
`For the first set of tests the coordination compound
`Calculated for C6H 10N20 8Pt C:l6.63 H 2.33 N:6.47;
`was freshly dissolved in sterile distilled water and in-
`Found C:16.60 H 2.64 N:6.80.
`jected intraperitoneally on day 1 into each of the test
`20 mice. The volume of the injection was usually ! ml. In
`GENERAL STRUCTURE CONFORMATION
`The malonate group is shown to be coordinated to
`some cases, in order to get an active dose into the ani-
`the platinum by the observed change in the electronic
`mal where the chemical was not soluble in this amount
`of solvent, a fine dispersion was prepared of the dose
`spectra on going from the aquo to the malonate species.
`Thus, structures such as [Pt(NH3)z(H20)2h(H2C30 4)]
`needed for the test. Thus, some of our test results were
`are ruled out confirming the analytical data. Similarly, 25 obtained on animals where a slurry of the compound
`zero-time conductivity measurements support a neutral
`was injected. These are so noted in Table I below. In
`compound. The i.r. spectra show the presence of coor-
`addition, for some of the compounds, there was injected
`dinated carboxyl groups (1600-1650 cm- 1 and 1400
`about 1 ml of solution, either in one single injection, or
`in 2 injections given a few hours apart of ! ml each.
`em- 1) with no C02H groups (which would show at
`1700-1750 em). Finally the carboxyl group vibrations 30 These injections were initially given in 4 different dose
`are compatible with a chelated structure as compared to
`levels for each new compound with 6 mice in each dose
`oxalate complexes of known structures.
`level. The tests covered a dose range from a low inef-
`The compounds of the invention were tested for
`fective dose, to an upper dose level which produced
`anti-tumor activity using our standard screening tumor,
`some deaths within the time period of the experiment.
`solid sarcoma 180 turner in female Swiss white mice, 35 The results are set forth in Table I.
`
`TABLE I
`Tests of Antitumor Activity of Malonato and Substituted Malonato
`Coordination Complexes of Platinum.
`
`Tumor-Sarcona 180
`Animal-Female Swiss white mice
`Single injection on days noted, intraperitonea11y
`
`Day of
`Injection
`
`Dose Level
`10 mglkg
`
`No. of
`TIC Deaths
`76 0
`
`Coordination Complex
`Malonatodiammineplatinum
`(II) (slurry in H20)
`
`(solution in H20)
`
`Daily for
`days 1-10
`
`Methylmalonatodiammine(cid:173)
`platinum(ll) (Solution in H20)
`
`malonatoethylenediamine(cid:173)
`platinum (II)
`
`ethylmalonatoethylenediamine(cid:173)
`platinum (II)
`
`malonato-1 ,2 propylenediamine(cid:173)
`platinum (II)
`
`15 mg/kg
`20 mglkg
`25 mg/kg
`30 mg/kg
`40 mg/kg
`50 mg!kg
`60 mg/kg
`4 mglkg
`
`5 mg/kg
`6 mglkg
`7 mg/kg
`
`30 mglkg
`40 mg/kg
`SO mglkg
`60 mglkg
`70 mg/kg
`80 mg/kg
`60
`80
`100
`120
`40
`60
`80
`90
`100
`110
`120
`45
`60
`75
`90
`
`38 0
`64 0
`31
`0
`7 1/6
`6!6
`5/6
`6/6
`54 0
`
`56 0
`23
`0
`12 0
`
`39 0
`26 0
`35
`0
`6 0
`124 3/6
`6/6
`80 0
`138 0
`85
`0
`50 0
`72 0
`81
`0
`79
`0
`47
`0
`I
`55
`41
`0
`58
`0
`50 0
`I
`9
`16
`3
`5
`
`NEPTUNE GENERICS 1006-00004
`
`APOTEX 1 006 - 0004
`
`

`
`7
`-continued
`Day of
`Injection
`
`Coordination Complex
`malonato-1,3 propylenediamine-
`platinum (II)
`
`methylmalonatoethylene-
`diamineplatinum (II)
`(solution in H20)
`
`ethylmalontodiammine-
`platinum(II)
`(solution in H20)
`
`malonatoethylenediamine-
`platinum (II)
`(solution in H20)
`
`l,l-cyclobutanedicarboxylate
`diammineplatinum (II)
`
`malonatobis(methylamine)
`platinum (II)
`
`4,140,707
`
`8
`
`Dose Level
`20
`40
`60
`80
`30 mg/kg
`
`40 mg/kg
`50 mg/kg
`60 mg/kg
`70 mg/kg
`90 mg/kg
`30 mg/kg
`
`40 mg/kg
`50 mg/kg
`60 mg/kg
`70 mg/kg
`80 mg/kg
`10 mg/kg
`
`20 mg/kg
`40 mg/kg
`45 mg/kg
`50 mg/kg
`55 mg!kg
`60 mg/kg
`80 mg/kg
`20 mg!kg
`
`40 mg/kg
`60 mg/kg
`80 mg/kg
`100 mg!kg
`120 mg!kg
`160 mg/kg
`80 mg!kg
`100 mg/kg
`120 mg/kg
`140 mglkg
`160 mg/kg
`180 mg/kg
`
`No. of
`Deaths
`0
`0
`0
`I
`0
`
`T/C
`69
`79
`21
`35
`78
`
`80
`51
`26
`20
`4
`57
`
`43
`47
`39
`17
`16
`88
`
`58
`18
`49
`35
`38
`15
`24
`71
`
`60
`38
`42
`69
`18
`62
`58
`53
`28
`25
`17
`19
`
`0
`0
`0
`I
`I
`0
`
`0
`0
`0
`0
`0
`0
`
`0
`0
`0
`0
`0
`3/6
`3/6
`0
`
`0
`0
`0
`0
`0
`4
`0
`0
`0
`0
`I
`I
`
`In addition to the day 1 injections described above, in 35
`a number of cases injections were delayed until day 8 of
`tumor growth. In these cases the tumor was usually at
`least larger than l gm, as estimated by palpation. The
`animals were then injected and observed for a period of
`approximately 60 days. Activity was measured by the 40
`number of animals whose tumors had regressed to the
`vanishing point, while still allowing the animal to sur(cid:173)
`vive for this time period. Such test results are described
`in TABLE II below.
`TABLE II
`Tests of Large Sarcoma 180 Regressions by Malonate
`Coordination Complexes of Platinum.
`Animal-Female Swiss white mice
`Tumor-Sarcoma 180
`Single injections on
`Day 8 intraperitoneally in H.O solutions
`
`45
`
`50
`
`TABLE III
`Confirmatory Tests of Antitumor Activity
`Malonatodiammineplatinum(II)
`Tumor: Walker 256 Carcinosarcoma- Animal- Rat
`Single injection Day I in Oil, Intraperitoneally
`Dose
`% Inhibition
`10 mg/kg
`100
`20 mg/kg
`100
`40 mg/kg
`100
`80 mg/kg
`-
`Malonatoethylenediamineplatinum(II)
`Tumor: Walker 256 Carcinosarcoma- Animal- Rat
`Single injection Day I in Oil, Intraperitoneally
`% Inhibition
`Dose
`
`Deaths
`0
`0
`0
`0
`
`Deaths
`0
`0
`0
`0
`all
`
`Dose
`14mg/kg
`
`!6mg/kg
`18 mg/kg
`20mg/kg
`40mg/kg
`
`Total Number
`of Regressions Deaths
`4
`2
`
`3
`4
`5
`3
`
`3
`2
`I
`3
`
`Coordination Complex
`malonatodiammine-
`platinum(II)
`
`malonatoethylene-
`diamineplatinum(II)
`
`I
`2
`3
`
`5
`4
`3
`
`45 mg/kg
`50 mg/kg
`60 mg/kg
`The results described in Tabl~s I and II indicate that
`the compounds of the invention are very potent antitu(cid:173)
`mor agents against the S 180 tumor in Swiss white mice.
`Confirmatory tests of antitumor activity against the
`Walker 256 Circinosarcoma in rats, and the ADJ/P(cid:173)
`C6A tumor in mice were conducted. The initial test
`results are shown in Table III and confirm the potent
`action of the compounds of the invention against these
`other tumor systems.
`
`55
`
`60
`
`65
`
`4
`20
`100
`500
`
`1.3
`94
`100
`
`Deaths
`0
`0
`0
`all
`
`Samples of the malonato diammine and malonato
`ethylene diamino complexes of platinum(II) were sub(cid:173)
`mitted to the Drug Research and Development Branch
`of the National Cancer Institute for screening for antitu(cid:173)
`mor activity against the L1210 tumor in mice. There(cid:173)
`sults obtained on this tumor system are shown in Table
`IV. They confirm the activity of the compounds of the
`invention.
`
`TABLE IV
`Confirmatory Tests of Antitumor Activity at the
`National Cancer Institute.
`Animal- Mice
`Tumor: L1210
`Daily injectons Days 1-9, Intraperitoneally
`
`NEPTUNE GENERICS 1006-00005
`
`APOTEX 1 006 - 0005
`
`

`
`Coordination Complex
`Malonatodiammineplatinum(II)
`
`Malonatoethylenediamineplatinum(ll}
`
`Dose
`50 mglkg
`25 mglkg
`12.5 mg/kg
`
`50 mg/kg
`25 mglkg
`12.5 mglkg
`37.5 mglkg
`(repeat test}
`Animal- Mice
`Tumor: Ll210
`Daily injectons Days 1-9, Intraperitoneally
`
`%Increase
`in Lifespan
`163
`133
`ll5
`lot
`160
`151
`121
`
`%Increase
`in Lifespan
`196
`160
`145
`
`5
`
`10
`
`IS
`
`Coordination Complex
`
`Dose
`25 mg/kg
`16.5 mg/kg
`II mg/kg
`
`The malonatoplatinum coordination compounds of
`the invention are preferably dissolved or suspended in 20
`water or other pharmaceutically acceptable carrier
`liquids. The parenterally administerable composition
`should preferably contain from about O.Smg to about
`lOmg per mi., it being understood that the amount may
`vary greatly depending upon the particular compound 25
`employed and the animal to be treated.
`The platinum coordination compounds of the inven(cid:173)
`tion are preferably administered parenterally to an ani(cid:173)
`mal affected with a malignant tumor. The duration of
`treatment and the dose level, of course, will depend in 30
`each case upon the size of the host animal, nature and
`size of the tumor, etc. Generally, however, a dose level
`of from about 20 to about 200 mg!kg of body weight
`per day will be sufficient. It is to be understood, how(cid:173)
`ever, that the platinum coordination compounds com- 35
`pounded with a suitable pharmaceutical carrier in the
`same proportions as recited above may also be adminis(cid:173)
`tered orally at the same dosage levels.
`
`40
`
`45
`
`50
`
`ss
`
`60
`
`65
`
`9
`TABLE IV-continued
`Confirmatory Tests of Antitumor Activity at the
`National Cancer Institute.
`
`4,140,707
`
`10
`
`We claim:
`1. Platinum coordination compounds having the for(cid:173)
`mula:
`[Pt(II)Ax((OOC)z-CRR 1ll
`
`wherein:
`x = 1 or 2;
`R and R 1 are selected from the group consisting of H,
`lower alkyl, aryl, aralkyl, alkenyl, cycloalkyl, cy(cid:173)
`cloalkenyl, alkoxy, OH, or combine with the car(cid:173)
`bon atom to form a cycloalkyl or cycloalkenyl
`group;
`when x = 1, A is HR2N-CHR3-CHR4-NRsH
`and when x = 2, A is H2NR6 or an amino acid,
`wherein Rz, R3, ~and R 5 are the same or different
`and are selected from the group consisting of H,
`CH3, C 2H 5, hydroxy and lower alkoxy, provided
`that R2 and R5 may also be aryl or aralkyl and each
`R6 is the same or different and is selected from the
`group consisting of H, lower alkyl, aryl, aralkyl,
`hydroxy lower alkyl, hydroxyl- and alkoxylamines,
`and alkoxyl alkyl amines.
`2. The compound of claim 1 having the formula:
`
`[Pt(Il)Ax((00Ch-CH2)]
`
`wherein:
`x = 1, and
`R2, R3, ~and R5 are each H.
`3. The compound of claim 1 having the formula:
`
`[Pt(II}Ax((OOCh-CH2)]
`
`wherein:
`x = 2, and each R6 is H.
`4. Malonate diammine platinum (II).
`
`. . "' . .
`
`NEPTUNE GENERICS 1006-00006
`
`APOTEX 1 006 - 0006
`
`

`
`;
`
`UNITED STATES PATENT AND TRADEMARK OFFICE
`
`CERTIFICATE EXTENDING PATENT TERM
`UNDER 35 U.S.C. 156
`
`Patent No.
`
`4,140,707
`
`Dated
`
`February 20, 1979
`
`Inventor(s)
`
`Michael J. Cleare et al
`
`Patent owner
`
`Research Corporation Technologies, Inc.
`
`This is to certify that there has been presented to the
`
`COMMISSIONER OF PATENTS AND TRADEMARKS
`an application under 35 u.s.c. 156 for an extension of the
`patent term. Since it appears that the requirements of the law
`have been met, this certificate extends the term of the patent
`for the period of
`
`with all rights pertaining thereto as provided by 35
`usc 156 (b) •
`
`916 DAYS
`
`I have caused the seal of the Patent
`and Trademark Office to be affixed
`this 25th day of January 1990.
`
`{7~~ f<-L~/
`
`Jeffrey M. Samuels
`Acting Commissioner of
`Patents and Trademarks
`
`NEPTUNE GENERICS 1006-00007
`
`APOTEX 1 006 - 0007
`
`

`
`REEXAMINATION CER1'IFICATE (lt73rd)
`United States Patent £191
`[111 Bt 4,140,707
`[4SJ Certificate ISsued Dec. 19, 1989
`
`[S4]
`
`(7S)
`
`Inventors: Michael J. Cleare; James D.
`Heescbeler, BlrBett ROHabeq;
`Loretta L Vaa Camp. aU of East
`Lansing. Mich.
`
`(73] Assignee: Research Corporatfoa Teebnologies,
`Inc., Tucson, Ariz.
`
`Reeulllfutloa Request:
`No. 90/001,?16, Feb. 14, 1989
`
`Reen•l11tloa Certlfkate for.
`Patent No.:
`4,140,707
`Issued:
`Felt. 20, 1979
`AppL No.:
`778,955
`Mar. 18, 1977
`Filed:
`
`Related U.S. AppUcatfoa Data
`(63} Continuation of Ser. No. 260,989, Jun. 3, 1972, aban·
`doned.
`Int. Q.4 .............................................. C07F 15/00
`[S1]
`[52] u.s. a. ........................................ 556/137; 546/2;
`SS6/l1
`[58] Field of Search ....................... 546/2; 5!!6/17, 137
`Relereac:e~~ Qted
`{56]
`PUBLICATIONS
`"On the Stereochemistry of Plato Salts (IV)", authored
`by A. A. Grunberg on Jan. 8, 1931 and published on
`May 2, 1931 in Helvetica Chimica Acta XIV at pp.
`4!55-472.
`.
`Primary Examiner-A. McFarlane
`ABS:fRACI'
`[!!7]
`Malonato platinum coordination compounds and a
`method of treating malignant tumors comprising the
`parenteral administration to an affected animal of a
`solution of the compound.
`
`NEPTUNE GENERICS 1006-00008
`
`APOTEX 1 006 - 0008
`
`

`
`Bl 4,140,707
`
`1
`REEXAMINATION CERTIFICATE
`ISSUED(:UNDER 35 u.s.c. 307
`
`!-
`·,' ~ .
`THE PATENT IS HEREBY AMENDED AS
`INDICA TED BELOW.
`
`wherein:
`x=lor2;
`s R and Rt are selected from the group consisting of H,
`lower alkyl, aryl, aralkyl, alkenyl, cycloalkyl, cy·
`cloalkenyl, alkoxy, OH. or combine with the car(cid:173)
`bon atom in CRR 1 to form a cycloalkyl or cy-
`cloalkenyl group;
`when x= 1, A is HR2N-CHR3-CHlt4-NR5H and
`when x=2, A is H2~or an amino acid, wherein
`R2, R3, R4 and R, are the same or different and arc
`selected from the group conSisting of H. CH3,
`C2H, hydroxy and lower alkoxy, provided that
`R2 and R, [may also be] are also Rlected from the
`group consisting of aryl or aralkyl and each R6 is the
`same or different and is selected from the group
`consisting of H. lower alkyl. aryl, aralkyl. hydroxy
`lower alkyl, hydroxyl- and alkoxylamines, and
`alkoxyl alkyl amines. provided that when x=2 and A
`is HzNR6 and R4 is H. then Rand R tare not both H.
`.Claim Z. dependent on an amended claim. is deter-
`5. The compound ofc/4im 1 wherein Rand Ri taken
`together wi'th the carbon to which they are attached form a
`mmcd to be patentable.
`cycloalkyf group.
`New claims 5, 6, and 7 arc added and determined to 25 6. The compound of claim 5 wherein R and R 1 taken
`be patentable.
`together wi'th the carbon atom to which they are attached
`form a cyclobutane.
`7. 1,1 cyclobutane dicarboxylate diammine platinum
`(II).
`• • • • •
`
`Matter enclosed in heavy bl1lCkets [] appeared in the
`patent, but hu been deleted and is no longer a part of the 10
`patent; matter printed lD italics Indicates additions DWle
`to the patent.
`
`AS A RESULT OF REEXAMINATION, IT HAS
`BEEN DETERMINED THAT:
`
`15
`
`Claims 3 and 4 are cancelled.
`
`Claim 1 is determined to be patentable as amended.
`
`1. Platinum coordination compounds having the for(cid:173)
`mula:
`
`20
`
`30
`
`3'
`
`40
`
`4S
`
`so
`
`ss
`
`60
`
`6S
`
`NEPTUNE GENERICS 1006-00009
`
`APOTEX 1 006 - 0009