United States Patent [191
`Cleare et a1.
`
`[11]
`[45]
`
`4,140,707
`Feb. 20, 1979
`
`[54] MALONATO PLATINUM ANTI-TUMOR
`COMPOUNDS
`[75] Inventors: Michael J‘ Clem-e; James D‘
`Hoeschele; Barnett Rosenberg;
`Lo
`L v
`81] f Eas
`L mt.”
`a; Camp’
`0
`t
`ansmg’ 1c '
`[73] Assignee: Research Corporation, New York,
`N-Y-
`
`[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 eta1., J. Pharmaceutical Sciences, 65, No. 3 (1976),
`pp. 315-320.
`Rosenberg et al., Nature 222, 385-386 (1969).
`
`[21] Appl' No‘: 778’955
`[22] Filed:
`Mar. 18, 1977
`
`Primary Examiner-Helen M. S. Sneed
`Attorney, Agent, or Firm-Dennis P. Clarke
`
`[63]
`
`Related US. Application Data
`Continuation of Ser. No. 260,989, Jun. 8, 1972,
`abandoned.
`[51] Int. 01.2 ............................................ .. C07F 15/00
`[52] US. Cl. .............................. .. 260/429 R; 424/245;
`
`ABSTRACT
`[57]
`Malonato Platinum coordination compounds and a
`method of treating malignant tumors comprising the
`foalret‘ftemlf :gmm‘s‘mm’; t° an affected
`°f ‘’
`“ ‘°“ ° ° °°mp°un -
`
`424/287; 546/4
`[58] Field of Search .................................. .. 260/429 R
`
`'
`
`-
`4 Claims, No Drawings
`
`Wockhardt Exhibit 1006 - 1
`
`

`
`1
`
`MALONATO PLATINUM ANTI-TUMOR
`COMPOUNDS
`
`The invention described herein was made in the
`course of work under a grant or award from the De
`partment of Health, Education and Welfare. -
`This is a continuation, of application Ser. No.
`260,989, ?led June 8, 1972, now abandoned.
`
`BACKGROUND OF THE INVENTION
`The present invention relates to novel malonato plati
`num coordination compounds and to their use in cancer
`chemotherapy.
`SUMMARY OF THE INVENTION
`The invention provides platinum coordination com
`pounds having the formula:
`
`10
`
`15
`
`20
`
`25
`
`30
`
`4, 140,707
`2
`num (IV) forms-dzsp3 coordination compounds which
`have an octahedral arrangement in space.
`The coordination compounds of the-invention in
`clude the cis and trans isomers of platinum (II) and
`platinum (IV) which contain the bidentate malonato
`ligand which may be substituted or'unsubstituted. The
`malonato ligand may contain substituents selected from
`the group consisting of lower alkyl, (e.g., methyl, ethyl,
`n-propyl, isopropyl, n-butyl, etc.); aryl, (e.g., phenyl;
`lower alkyl-, lower alkenyl-, halo-', nitro-, lower alkoxy
`substituted phenyl and naphthyl); aralkyl, (e. g., phenyl
`methyl (benzyl), 2-(l-naphthyl)methyl); alkenyl, (e.g.,
`4-amino-l-butene, allyl); cycloalkyl, (e.g., cyclopropyl,
`cyclohexyl, etc.); cycloalkenyl, (e.g., 2-cyclopenten
`l-yl, 2-cyclohexen-l-yl); alkoxy, (e.g., methoxy, ethoxy,
`etc.), and hydroxy. Also suitable are the ‘Ll-cycloalk
`ylenedicarboxylic
`acids,
`(e. g.,
`l,l-cyc1o
`propanedicarobxylic acid, l,l-cyclobutanedicarboxylic
`acid, etc.) and the l,l-cycloalkenyldicarboxylic acids,
`(e.g.,
`l, l-cyclopropenedicarboxylic
`acid,
`1, 1
`cyclobutenedicarboxylic acid, etc.)
`1
`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
`is 2 or one bidentate amine ligand, i.e., when x is 1.
`Suitable monodentate amine ligands include lower
`alkyl amines, (e.g., methyl-, ethyl-, n-propyl-, isopropyl
`, n-butyl- amines, etc.), aryl amines, (e.g., aniline), aral
`kyl amines, (e.g., benzylamine), hydroxy lower alkyl
`amines, (e.g., ethanolamine, propanolamine, etc), hy
`droxylamine, lower alkoxy amines (e.g., methoxyla
`mine, etc.), alkoxyalkylamines (e.g., methoxymethyla
`mine, etc.), and heterocyclic amines (e.g., pyridine and
`aziridine). Also included are the amino acids, i.e.,
`R7-—CHNH2-—COOH wherein R7 is H, lower alkyl
`(e.g., methyl, isopropyl, etc.), hydroxy lower alkyl (e.g.,
`hydroxymethyl, hydroxyethyl, etc.), aralkyl (e.g., ben
`zyl, etc).
`It is to be understood that the coordination com
`pounds of the invention may include two identical or
`different monodentate ligands.
`Suitable bidentate amine ligands include the substi
`tuted and unsubstituted primary and secondary
`ethylenediamines. One or both of the carbon atoms of
`the ethylenediarnine may contain substituents such as
`lower alkyl (e.g., methyl, ethyl), hydroxyl, alkoxy (e.g.,
`methoxy, ethoxy, etc). Secondary ethylenediamines
`wherein one or more of the amine groups contains sub
`stituents such as listed above for the carbon atoms of the
`primary amine and aryl (e.g., phenyl) and aralkyl (, e. g.
`benzyl) may also be utilized.
`The Pt (II) coordination compounds specified herein
`do not exist as geometrical isomers; however, the Pt
`(IV) compounds exist as cis and trans isomers. It is to be
`further understood that the invention is inclusive of the
`cis and trans ‘isomers.
`.
`The Pt (IV) coordination compounds may also con
`tain two monodentate or one bidentate anionic ligand
`where only one malonato ligand is present, i.e., where y
`= l in the above formula.
`Suitable monodentate anionic ligands include chlo
`ride, bromide, iodide, nitrite-hydroxide, nitrate, sulfa
`mate, etc. Among the bidentate anionic ligands which
`may be present are oxalate, pyrophosphate, dithioxa
`
`wherein:
`x = l or 2;
`y = l or 2:
`z = 0, l or 2,
`provided that when y = 2, z = 0 and when y = 1, z is
`greater than 0;
`R and R1 are selected from the group consisting of H,
`lower alkyl, aryl, aralkyl, alkenyl, cycloalkyl, cy
`cloalkenyl, alkoxy, OH, or are combined with the car
`bon atom to form a cycloalkyl or cycloalkenyl group,
`and substituted derivatives thereof;
`35
`when x
`l, A is HR2N—CHR3—CHR4—NR5H
`and when x = 2, A is H2NR6a heterocyclic amine or an
`amino acid, wherein R2, R3, R4 and R5 are the same or
`different and are selected from the group consisting of
`H, CH3, C2H5, 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 consist
`ing of H, lower alkyl, aryl, aralkyl, hydroxy lower
`alkyl, hydroxyl and alkoxyl amines, alkoxylalkylamines
`wherein all of said alkyl groups are lower alkyls and
`heterocyclic substituents including said N as a ring
`member;
`.
`when 2 = l, L is a bidentate anionic ligand, and
`when 2 = 2, L is a monodentate anionic ligand.
`The invention also relates to a composition and
`method for treating malignant tumors in animals com
`prising parenterally administering to an animal affected
`with a malignant tumor a solution containing a platinum
`coordination compound as de?ned hereinabove in an
`amount sufficient to cause regression of the tumor.
`
`40
`
`45
`
`50
`
`55
`
`DETAILED DESCRIPTION OF THE
`INVENTION
`Platinum coordination compounds and methods for
`their production are described by J. C. Bailar, Jr., The
`Chemistry of the Coordination Compounds, Reinhold
`Publishing Corp., N.Y., 1956, Chap. 2; J. Lewis et al,
`Modern Coordination Chemistry: Princi'pIesand Methods,
`lnterscience Publishers, Inc., N.Y., 1960 and Kauffman
`Inorganic Synthesis, 7, McGraw-Hill Book Co., Inc.,
`N.Y., 1963.
`Platinum (II) forms dsp2 coordination compounds
`which have a square planar arrangement in space. Plati
`
`65
`
`late.
`
`'
`
`.
`
`It is to be understood that the invention includes
`those coordination compounds containing mixed mono
`dentate anionic ligands,
`
`1
`
`Wockhardt Exhibit 1006 - 2
`
`

`
`3
`The preferred compounds are those wherein R and
`R1 in the above formula are H, methyl or ethyl, i.e.,
`malonatoplatinum, methylmalonatoplatinum and ethyl
`malonatoplatinum coordination compounds. The most
`preferred Pt (II) compounds are those malonato
`platinum (II) compounds of the above formula wherein
`x = l and R2, R3, R4and R5 are each H, i.e., malonatoe
`thylenediamine platinum (II),
`methylmalonatoe
`thylenediamineplatinum (II) and ethylmalonatoe
`thylenediamineplatinum (II); and wherein x = 2 and
`each R6 is H, i.e., .malonatodiammineplatinum (II), me
`thylmalonatodiammineplatinum
`(II)
`and
`ethyl
`malonatodiammineplatinum (II).
`The preferred Pt (IV) compounds are those wherein
`x = 2, each R6 is H and y = 2, i.e., bismalonato (or
`bismethylmalonato or bisethylmalonato) diammine plat
`inum (IV).
`The coordination compounds of the invention may be
`prepared by one of a variety of well-known methods. A
`general method of preparation of the Pt (II) coordina
`20
`tion compounds is as follows: Starting compounds hav
`ing the formula cis-[Pt A(Hal)2] wherein Hal is 1, C1 or
`Br and A is one bidentate or two monodentate amine
`ligands (prepared by the method of S. C. Dhara, Indian
`J. Chem., Vol 8, p. 193 (1970)) are reacted with silver
`nitrate to form the diaquo complex. The latter is then
`reacted with the malonate ion to form the coordination
`compounds of the invention. This method is repre
`sented by the following reaction scheme:
`
`30
`
`35
`
`wherein A is one bidentate amine ligand or two mono
`dentate amine ligands.
`The following non-limiting examples are illustrative
`of the methods for preparing the compounds of the
`invention.
`
`EXAMPLE 1
`Malonatodiammineplatinumal)
`[PtmH3)2(C3H2O4)]
`
`Reactions:
`
`[Pt(NH3)z(H20)?(N0s)2 + CsHz04Z.— —>
`[mmnpgqnzonl + 2N0; * +2n2o.
`
`50
`
`11
`
`55
`
`Silver nitrate (22.55g — slightly less than the stoi
`chiometric amount in order to avoid silver contamina
`tion) was dissolved in water (50 ml.) and added to
`[Pt(NH3)2Cl2] (20g) in a 250 ml. conical ?ask. The con
`tents were warmed (60° C.) on a hot plate with rapid
`stirring until the silver chloride precipitation was com
`plete and the mother liquor was almost colorless. The
`silver chloride was ?ltered off using a ?ne pore sintered
`glass ?lter and the precipitate was washed several times
`with hot water to give a total ?ltrate volume of 100-200
`ml.
`Malonic acid (13g — a twofold excess) was dissolved
`in water (30 ml.) and neutralized with a solution of 65
`KOH (~ 13g in 30 ml.) to pH 5-6. The resulting potas
`sium malonate solution was added to the platinum con
`taining ?ltrate and the mixture was carefully warmed
`
`4,140,707 ~
`
`4
`(to avoid “bumping”) on the hot plate until white crys
`tals of the product started to form in’ great quantity. The
`mixture was then cooled 'to, room temperature and the
`product ?ltered off. The ?ltrate was reheated for 5-10
`minutes and cooled to 0° C. to collect a further crop.
`The crude yield at this stage was 20.5g (93%).
`The product was recrystallized by dissolving in boil
`ing or near boiling ‘water. The above yield (20.5g) re
`quired about 3 liters of boiling water for complete disso
`lution. Malonic acid lg/L was dissolved in the water to
`suppress any hydrolysis.‘ The ?ltered solution was
`cooled to. 0° C. to give white fluffy needles
`
`(l8.25g-83%). -
`.
`'U.V./via spectral and conductivity studies have shown that hydrolysis
`is negligible.
`The crystals decompose between l85°—l90° C. The
`structure of the product was veri?ed cia an i.r. spec
`trum. Solubility of the product is low in cold water, i.e.,
`20 mg/ 100 mls at 20° C. and 43 mg/ 100 mls at 37° C.,
`but higher in near boiling water (90°-l00° C.)~65g/ 100
`ml.
`The empirical composition was veri?ed by elemental
`analysis:
`Malonatodiammineplatinum?l) [Pt(NH3)2(C3H2O4)]
`Calculated for C3H8N2O4Pt.C.: 10.88; H: 2.43; N:
`8.46: Pt 58.9; Found C: 10.67; H: 2.35; N: 8.54; Pt 58.7.
`
`EXAMPLE 2
`[Pt (en) (C4H404)] (en = H2N(CH2)2NH2; C4H404 _
`= 02c omens) 00221
`Silver nitrate (3.64g) was dissolved in 20 ml of water
`and added to [Pt(NH2)2(CH2)2CL2] (3.5g) suspended in
`water (30 ml.) in a conical ?ask. The mixture was stirred
`on a warm hot plate for 5_—l0 minutes until all the yel
`low platinum complex had dissolved to give a yellow
`liquor plus a copious white silver chloride precipitate.
`The mixture was ?ltered through a ?ne pore ?lter and
`the precipitate washed twice with small volumes of hot
`water. The clear ?ltrate plus washings was added to an
`aqueous solution of methylmalonic acid (2g in 20mls)
`which had been adjusted‘ to pH 5-6. The mixture was
`heated to‘ about 80° C. for ?ve minutes and then cooled
`to 01C. The shiny white crystals which formed were
`?ltered and washed with cold water and acetone (Yield
`2.65g). The mother liquor plus aqueous washings was
`reduced to about half its original volume (~30 mls) to
`yield a second crop on cooling to 0'’ C. (Yield 0.85g).
`Total Crude yield was 3.50 'gms'(88%). The complex
`was recrystallized from a minimum volume of boiling
`water (around 250 mls) with ?ltration through a ?ne
`pore ?lter prior to cooling to 0° C.
`Yield of ‘shiny white lea?ets 2.96g (74%).
`Calculated for C6H12N2O4Pt; C:19.4l H 3.26 N:7.55;
`Found C:l9.ll H 3.61 N_:7.89.
`A second crop (0.33g—8%) was obtained by reducing
`the bulk of the mother liquor. -
`
`EXAMPLE 3
`
`Halls-[PI IV(NH3)2‘(ma1)2]
`_
`.
`Silver nitrate (5.45g) was dissolved in water (30 ml)
`and added to trans [Pt(NH3)2Cl4] (3g) suspended in
`water (30 mls) containingconcentrated nitric acid (3
`ml).- Thecontents were warmed on a hot plate (70°—80°
`C.) and stirred for‘at least one hour. The mixture was
`?ltered through a ?ne pore sintered glass ?lter to re
`move the silver chloride. The precipitate was washed
`twice with a small volume of hot water. The clear ?l
`
`Wockhardt Exhibit 1006 - 3
`
`

`
`0
`
`5
`
`4, 140,707
`6
`5
`following standard protocols for this testing as set by
`[rate 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(l962)).
`ent. (If the test is positive, suf?cient 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 sacri?ced
`cipitated.) The solution was re?ltered and the ?ltrate
`mouse was disected free of superfluous tissue and out
`under sterile conditions into approximately 10 milli
`reduced to 20-30 mls in volume and cooled to 0° C. to
`yield
`plate yellow crystals (presumably trans
`gram size pieces. These tissue pieces were then im
`planted by trocar in the left axillary region, subcutane
`(Pt(NH_~,)2(NO3)4]). These were washed with a little
`cold water and then acetone (Yield 1.8g). A portion of
`ously, in new mice. The mice were, on the average,
`approximately four weeks old and weighed 18-20
`this yield (lg) was dissolved in a minimum of hot water
`grams. Taking day 0 as the day of implant, the animals
`to which sodium nitrate (0.2g) had been added. This
`solution was ?ltered into an aqueous solution of malonic
`were sacri?ced on day 10. The tumors were excised and
`weighed and the ratio of the weights of the tumors in
`acid (0.5g — a slight excess) which had been adjusted to
`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 ?l
`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.
`For the ?rst set of tests the coordination compound
`(Yield 0.7g — 30-40%).
`Calculated for C6H10N2O8Pt C:l6.63 H 2.33 N:6.47;
`was freshly dissolved in sterile distilled water and in
`jected intraperitoneally on day 1 into each of the test
`Found C:l6.60 H 2.64 N:6.80.
`mice. The volume of the injection was usually 5 ml. In
`GENERAL STRUCTURE CONFORMATION
`some cases, in order to get an active dose into the ani
`The malonate group is shown to be coordinated to
`mal where the chemical was not soluble in this amount
`the platinum by the observed change in the electronic
`of solvent, a ?ne dispersion was prepared of the dose
`spectra on going from the aquo to the malonate species.
`needed for the test. Thus, some of our test results were
`Thus, structures such as [Pt(NI-I3)2(H2O)2]2(H2C3O4)]
`obtained on animals where a slurry of the compound
`are ruled out con?rming the analytical data. Similarly,
`was injected. These are so noted in Table I below. In
`zero-time conductivity measurements support a neutral
`addition, for some of the compounds, there was injected
`compound. The in spectra show the presence of coor
`about 1 m1 of solution, either in one single injection, or
`dinated carboxyl groups (1600-1650 cm_1 and 1400
`in 2 injections given a few hours apart of 21, ml each.
`cm“) with no COZH groups (which would show at
`These injections were initially given in 4 different dose
`1700-1750 cm). Finally the carboxyl group vibrations
`levels for each new compound with 6 mice in each dose
`are compatible with a chelated structure as compared to
`level. The tests covered a dose range from a low inef
`oxalate complexes of known structures.
`fective dose, to an upper dose level which produced
`The compounds of the invention were tested for
`some deaths within the time period of the experiment.
`anti-tumor activity using our standard screening tumor,
`The results are set forth in Table I.
`solid sarcoma 180 turner in female Swiss white mice,
`
`35
`
`TABLE I
`Tests of Antitumor Activity of Malonato and Substituted Malonato
`Coordination Complexes of Platinum.
`
`Animal-Female Swiss white mice
`Tumor - Sarcona 180
`Single injection on days noted, intraperitoneally
`
`Coordination Complex
`Malonatodiammineplatinum
`(II) (slurry in H20)
`
`Day of
`Injection
`l
`
`Dose Level
`10 mg/kg
`
`No. of
`T/C Deaths
`76 0
`
`(solution in H20)
`
`Daily for
`days 1-10
`
`Methylmalllonastoldiammine]:I O)
`latinum
`o ution in
`p
`( )(
`2
`
`malonatoethylenediamine-
`platinum (II)
`
`ethylmalonatoethylenediamine-
`platinum (II)
`~
`
`malonato-1,2 propylenediamine-
`platinum (ll)
`
`1
`
`l
`
`l
`
`l
`
`15 mg/kg
`20 mg/kg
`25 mg/kg
`30 mg/kg
`40 mg/kg
`50 mg/kg
`60 mg/kg
`4 mg/kg
`
`2
`m g
`7 mg/kg
`3O g/k
`m g
`40 mg/kg
`50 mg/kg
`60 mg/kg
`70 mg/kg
`80 mg/kg
`60
`80
`100
`120
`40
`60
`80
`90
`100
`110
`120
`45
`60
`75
`9O
`
`38 0
`64 0
`31 0
`7 V6
`— 6/6
`1
`5/6
`— 6/6
`54 0
`
`22 s
`12 O
`39 o
`26 0
`35
`0
`6 0
`124 3/6
`— 6/6
`80 0
`138 O
`85 O
`50 0
`72 0
`81
`0
`79 0
`47 0
`55
`1
`41 0
`58 0
`50 0
`9
`l
`16
`3
`— 5
`
`Wockhardt Exhibit 1006 - 4
`
`

`
`7
`-cont1nued
`Day of
`Injection
`l
`
`l
`
`l
`
`1
`
`l
`
`l
`
`Coordination Complex
`malonato-l,3 propylenediamine-
`platinum (II)
`
`methylmalonatoethylene-
`diamineplatinum (ll)
`(solution in H20)
`
`ethylmalontodiammine-
`platinum(ll)
`(solution in H20)
`
`malonatoethylenediamine-
`platinum (11)
`(solution in H20)
`
`l,l-cyclobutanedicarboxylate
`diammineplatinum (II)
`
`malonatobis(methylamine)
`platinum (II)
`1
`
`4,140,707
`
`8
`
`Dose Level
`20
`40
`6O
`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 mg/kg
`160 mg/kg
`180 mg/kg
`
`No. of
`T/C Deaths
`69 0
`79 0
`21
`0
`35
`l
`78 0
`
`80 0
`51 0
`26 0
`20
`l
`4 l
`57 0
`
`43 0
`47 0
`39 0
`17 0
`16 0
`88 0
`
`53 0
`18 0
`49 0
`35 0
`38 O
`l5
`3/6
`24 3/6
`71 0
`60 0
`38 O
`42 0
`69 O
`18 O
`62 4
`58 O
`53 0
`28 0
`25 0
`17
`l
`19
`1
`
`_
`
`In addition to the day 1 IHJCCUOHS described above, in 35
`a number of cases lnjectlOnS were delayed unt1l day 8 of
`tumor growth. In these cases the tumor was usually at
`-
`-
`least larger than 58.31, as estlmated 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 t e anima to sur
`vive for this time period. Such test results are described
`.
`m TABLE H below‘
`TABLE II
`Tests of Large Sarcoma 180 Regressions by Malonato
`Coordination Complexes of Platinum.
`Tumor-Sarcoma 180
`Animal-Female Swiss white mice
`Single injections on
`Day 8 intraperitoneally in H2O solutions
`
`TABLE III
`Con?rmatqry Tests of Amimmo, Activity
`lrhlcnatgvdialrnmggzpgtinumm)
`A I
`umor: 3 Cl‘
`arcmosarcoma- nima -Rat
`Single injection Day 1 in Oil, lntraperitoneally
`Dose
`% Inhibition
`10 mg/kg
`100
`20 mg/kg
`100
`80 mg/kg
`_
`¥alonat$tlglen2e€6iaglineplatinum(ll)
`umor: a er
`arcinosarcoma - Animal - Rat
`Single injection Day 1 in Oil, Intraperitoneally
`Dose
`% Inhibition
`m
`20
`40
`80
`160
`gumlorg ADJ/"3A -2_r.;\nin(i)all -IMouse
`in
`i , ntra ritonea
`mg e injectlon ay
`Dose
`% Inhibitig?
`4
`‘
`20
`~3
`100
`194
`500
`0°
`_'
`
`Deaths
`0
`0
`0
`
`0
`
`Deaths
`
`0
`0
`0
`23!
`
`n
`y Deaths
`
`0
`0
`0
`3"
`
`40
`
`l
`25
`
`_
`
`_
`
`45
`
`50
`
`55
`
`.
`
`.
`
`-
`
`.
`
`i
`
`-
`
`h
`
`.
`
`l
`
`_
`
`-
`~
`Coordination Complex
`malonatodiammine-
`platinum(l1)
`
`malonatoethylene-
`diamineplatinum?l)
`
`Dose
`14 mg/kg
`16 mg/kg
`18 mg/kg
`20 mg/kg
`40 mg/kg
`45 mg/kg
`2g 25::
`
`Total Number
`-
`of Regressions
`2
`3
`4
`5
`3
`1
`g
`
`Deaths
`4
`3
`2
`1
`3
`5
`‘;
`
`Samples of the malonato dlammme and malonato
`ethylene diamlno complexes of platinum(II) were sub
`mltted to the Drug Research and Development Branch
`The results described in Tables I and II indicate that 60 of the National cf‘mcer Instltute for “We”? for anmu
`.
`.
`-
`mor activity against the Ll2l0 tumor in mice. The re
`the compounds of the invention are very potent antltu-
`.
`.
`_
`-
`sults obtained on this tumor system are shown 11'! Table
`,
`.
`.
`.
`mor agents against the S 180 tumor in Swiss white mice.
`Iv The con?rm the activit of the com ound of th
`con?rmatory tests of antitumor activity against the
`inv'entio:
`y
`P
`S
`C
`Walker 256 Circinosarcoma in rats, and the ADJ/P- 65
`'
`TABLE IV
`C6A tumor in mice were conducted. The initial test
`Con?rmatory Tests of Antitumor Activity at the
`results are shown in Table III and con?rm the potent
`National Cancer Institute.
`action of the compounds of the invention against these
`Animal - Mice
`Tumor: Ll2l0
`Daily injectons Days l-9, Intraperitoneally
`other tumor systems.
`
`Wockhardt Exhibit 1006 - 5
`
`

`
`9
`TABLE IV-continued
`cmmma'm?azgi'zliga‘zgéf‘izmligmw a‘ 'he
`_
`h
`% Increase
`Coordination Complex
`in Lifespan
`Malonatodiammineplatinum?l)
`163
`l”
`
`Malonatoethylenediamineplatinum?l)
`
`. m
`
`Dose
`50 mg/kg
`ii §““"‘)i
`g g
`so mg/kg
`25 mg/kg
`mg/niig
`(repeal ‘e50
`Animal _ Mice
`Tumor: U210
`Daily injectons Days 1-9, lntraperitoneally
`
`Coordination Complex
`
`Dose
`25 Ins/kg
`16.5 mg/kg
`ll mg/kg
`
`% Increase
`in Lifespan
`196
`160
`I45
`
`4, 140,707
`
`10
`
`15
`
`101
`160
`
`We claim:
`'
`1. Platinum coordination compounds having the for
`mula:
`
`5
`
`)2_CRR|)]
`
`2
`
`'
`
`15
`
`[PtuDAxa
`h -
`2_
`w elfml
`X " or i
`_
`,
`-
`10 R and R1 are selected from the group consisting of H,
`lower alkyl, aryl, aralkyl, alkenyl, cycloalkyl, cy
`cloalkenyl, alkoxy, OH, or combine with the car
`,bon atom to form a cycloalkyl or cycloalkenyl
`group.
`.
`’
`when x = 1, A is HR2N--CHR3-—CHR4-NR5H
`and when x = 2, A is H2NR6 or an amino acid,
`wherein R2, R3, R4and R5 are the same or different
`and are selected from the group consisting of H,
`CH3, CZHS, hydroxy and lower alkoxy, provided
`that R; 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, my], “alkyl,
`hydmxy lower alkyl, hydroxyl_ and alkoxylamines,
`and alkoxyl alkyl amines.
`2. The compound of claim 1 having the formula:
`
`The malonatoplatinum coordination compounds of
`the invention are preferably dissolved or suspended in
`water or other pharmaceutically acceptable carrier
`liquids. The parenterally administerable composition
`should preferably contain from about 0.5mg to about
`10mg per ml., 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-
`tion are preferably administered parenterally to an ani
`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 suf?cient. It is to be understood, how
`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
`tered orally at the same dosage levels.
`
`[P:(Il)A,((00C)2-CH2)]
`
`wherein:
`x = l, and
`R2, R3, R4 and R5 are each H.
`3- The compound of claim 1 having the formula:
`
`wherein:
`x = 2, and each R6 is H.
`4. Malonato diammine platinum (II).
`$ $ $ ‘ i
`
`45
`
`50
`
`55
`
`65
`
`Wockhardt Exhibit 1006 - 6
`
`

`
`UNITED STATES PATENT AND TRADEMARK OFFICE
`
`CERTIFICATE EXTENDING PATENT TERM
`UNDER 35 U.S.C. 156
`
`Patent No.
`
`Dated
`
`:
`
`:
`
`4,140,707
`
`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.
`
`gyji? fix
`
`Jeffrey M. Samuels
`Acting Commissioner of
`Patents and Trademarks
`
`Wockhardt Exhibit 1006 - 7
`
`

`
`‘REEXAMINATION CERTIFICATE (1mm)
`United‘
`P81231111 [191
`[111 B1 4,140,707
`[45] Certi?cate lcsued
`Dec. ‘19, 1989
`
`‘ "PLATINUM ANTI-TUMOR
`
`(75] Inventors: Michael J. Clenre; James D.
`Heelcheler; Barnett Roeenberg;
`Lore“! L. Van Camp, all of East
`Lansing. Mich.
`
`[731 Assignee: Research Corporation Technologiec,
`Inc” Tucson, Ariz.
`Reeuguinntion Request:
`- No.‘90/00l.716. Feb. 14, 1989
`
`Reexamination Certi?cate for: ~
`Patent No.:
`4,140,707
`Issued:
`Feb. 20, 1919
`Appl. No.:
`778,955
`Filed:
`Mar. 18, 1977
`
`[63]
`
`Relnted [1.8. Application Data
`Continuation of Ser. No. 260,989, Jun. 3, I972, aban
`doned.
`
`[51] Int. CL‘ ............................................ .. ('JWF 15/00
`[52] US. CL ...................................... .. 556/137; 546/2;
`556/ I7
`[58] Field of Search ..................... .. 546/2; 556/ 17, 137
`[56]
`References Cited
`PUBLICATIONS
`“On the Stereochemisrry of Plato Salts 0V)", authored
`by A. A. Grunberg on Jan. 8, 1931 and published on
`May 2, 1931 in Helvetica Chimioa Acta XIV at pp.
`
`455-472.
`
`‘
`
`'
`
`Primary Examiner-A. McFarlnne
`[57]
`ABSTRACT
`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.
`
`'
`
`Wockhardt Exhibit 1006 - 8
`
`

`
`Bl 4,140,707
`2
`IPKIDMKOOQPCRRDI
`
`1
`REEXANQHA'I'ION CERTIFICATE‘
`ISSUEQIUNDER 35 U.S.C. 307
`THE PATENT IS HEREBY AMENDED AS
`INDICATED BELOW.
`
`AS A. RESULT OF REEXAMINATION, IT HAS
`BEEN DETERMINED THAT:
`
`Claims 3 and 4 ere cancelled.
`
`Claim 1 is determined to be patentnble as amended.
`
`Matter enclosed in heavy brackets [] lppeared in the
`patent, but has been deleted and is no longer a part of the
`potent; matter printed in italics indicates additions made
`to the patent.
`
`wherein:
`=l or 2;
`R and R1 are selected from the group consisting of H,
`lower alkyl, aryl, aralkyl, elkenyl. cycloalkyl. cy
`cloalkenyl, alkoxy, OH, or combine with the car
`bon atom in CRR] to form a cycloalkyl or cy
`cloalkenyl group;
`when x: l, A is HRzN-CHR3-CHR4-NR5H and
`when x=2, A is HzNR6 or an amino acid, wherein
`R2, R3, R4 and R5 are the same or different and are
`selected from the group consisting of H, CH3,
`CzH5, hydroxy and lower elkoxy, provided that
`R; and R5 [may also be] are also selected from the
`group consisting ofaryl or eralkyl and each R6 is the
`same or different and is selected from the group
`consisting of H, lower alkyl, eryl, arnlkyl, hydroxy
`lower alkyl, hydroxyl- and elkoxylamines, and
`alkoxyl elkyl amines. provided that when x=2 and A
`irHZNRG and R61‘: H, then R and R1 are not both H.
`5. The compound of claim‘ I wherein R and Ri taken
`Claim 2; dependent on an amended claim, is deter
`together with the carbon to which they are attached form 0
`mined to be patentable.
`loo] 1
`Newclsims5,6,nnd7areaddedanddeterminedto25cyc kygmp
`6. The compound of claim 5 wherein R and R1 taken
`together with the carbon atom to which they are attached
`be patentable.
`jbrm a cyclobutane.
`7. 1,1 cyclobutane dtbarboxylate diammine platinum
`
`1. Platinum coordination compounds having the for
`mule:
`
`30
`
`45
`
`55
`
`Wockhardt Exhibit 1006 - 9