ANTITUMOR PLATINUM COMPLEXES

Abstract: The present invention relates to platinum (II) complexes showing antitumor activity and having the formulae and wherein the stereoisomerism of 1,2-diaminocyclohexane is cis-, trans(+) or trans(-) and R¹ and R² when taken together represents a thymidylato-, phosphonoacetato-, or hydroxymethylphosphonato group. The invention also relates to a process for preparing these compounds and to pharmaceutical compositions containing such a complex.

Description: This invention relates to novel platinum (II) complexes having antitumor activity. More particularly, it relates to novel antitumor platinum (II) complexes of 1,2-diaminocyclohexane and 1-aminomethylcyclooctylamine. Various platinum (II) complexes of 1,2-diaminocyclohexane, 1-aminomethylcyclooctylamine and 1,2-diamino-2,4-dimethylpentane have been disclosed in the scientific and patent literature as having antitumor activity. With respect to platinum (II) complexes of 1,2-diaminocyclohexane, the original antitumor complex appears to have been dichloro(1,2-diaminocyclohexane)platinum (II) reported, for example, in Bioinorg. Chem. 2: 187-210 (1973). The chloride ligands in this complex were then replaced by various inorganic and organic ligands such as nitrato, methanesulfonato, tartrato, sulfato, methylmalonato, orthophosphato, acetylacetonato, pyruvato, phthalato, oxalato, glycerophosphato, gluconato, glycerato etc. (see, for example, Cancer Treat. Rep., 61(8): 1519 - 1525 (1977). These and other analogs are disclosed in U.S. Patent 4,169,846, J. Med. Chem. 21(12): 1315 - 1318 (1978), J. Clin. Hematol. Oncol. 7(4): 867-876 (1977) - see bis(monobromoacetato) analog, J. Clin. Hematol. Oncol. 7(1): 231 - 241 (1977); U.K. Patent Application 2,003,468A - see 4-carboxyphthalato analog, J. Clin. Hematol. Oncol. 7(1): 220 -230 (1977) - see analogs of ligands such as ketomalonate, 1,2-dibromomaleate, 1,2-dibromosuccinate, J. Clin. Hematol. Oncol. 8(2): 44 - 50 (1978) - note disclosure of analog of the formula U.S. Patent 4,115,418, U.S. Patent 4,256,652 and U.S. Patent 4,284,579 - note preparation of cis-dihydroxy(1,2-diaminocyclohexane)platinum (II) and analog with N-phosphonoacetyl-L-asparatato ligand. J.Inorg. Biochem. 11: 139 - 149 (1979) discloses preparation of 1-aminomethylcyclooctylamine and 1,2-diamino-2,4-dimethylpentane (starting materials for certain complexes of the present invention) as well as complexes of such amine ligands with ligands such as Cl⁻, SO₄ = , CH₃(CCOO⁻)₂ and (BrCH₂COO⁻)₂. U.S. Patent 4,359,425 discloses antitumor organo-platinum complexes of the general formula wherein X and Y each or taken together represent a mono- or bifunctional ligand selected from the group consisting of halogenato, nitrato, sulfonato, monocarboxylato, sulfato and dicarboxylato and each of n and m is an integer of 1 or 2. European Patent Application 55,300 discloses as antitumor agents platinum (II) complexes of the formula wherein -B-B- represents or in which R₁ and R₂ are the same or different and each is hydrogen, an alkyl group or an aryl group, and n, m and 1 are 0 or an integer of from 1 to 3, at least one of A₁ and A₂ is (hereinafter referred to as a ligand of D-gluconic acid) and the other is the ligand of D-gluconic acid, Cl⁻, Br⁻, I⁻, F⁻, XCH₂COO⁻ (in which X is a halogen atom), NO₃⁻, SO₄⁻², H₂PO₄⁻ or H₂O or, when taken together, A₁ and A₂ may form a ring together with Pt(II), in the latter case -A₁-A₂- being the ligand of D-gluconic acid. U.K. Patent Application 2,093,845 discloses as antitumor agents platinum (II) complexes of the formula wherein either R₁ and R₂ independently are hydrogen or a substituted or unsubstituted alkyl, cycloalkyl, aryl or aralkyl group or R₁ and R₂ when taken together may be a substituted or unsubstituted cycloalkyl group, R₃ and R₄ are each independently hydrogen or a substituted or unsubstituted alkyl, aryl or aralkyl group and X is a chlorine, bromine or iodine atom, a sulfate radical, a substituted or unsubstituted carboxylate radical and Y independently from X is a chlorine, bromine or iodine atom, a hydroxyl group, a nitrate group or a carboxylate group. U.K. Patent Application 2,024,823A discloses as antitumor agent platinum (II) complexes of the formula in which R₁ and R₂ are each independently a hydrogen atom or an optionally substituted alkyl, cycloalkyl, aryl or aralkyl group, or R₁ and R₂ together are an optionally substituted cycloalkyl group, R₃ and R₄ are each independently a hydrogen atom or an optionally substituted alkyl, aryl or aralkyl group, and X is an anionic group. The amine ligand 1,2-diaminocyclohexane may be used as a mixture of cis-, trans-d-and trans-l-isomers to produce the corresponding isomer mixture derived from such starting material. Alternatively and preferably, the 1,2-diaminocyclohexane may be separated into the individual cis-, trans-d- and trans-l-isomers by the procedure described, for example, in U.S. patent 4,169,846. While all of these individual isomers may be utilized to prepare the complexes of the present invention, the preferred starting material is the trans-l-isomer, hereinafter referred to below as the trans(-)-isomer. This isomer has previously been reported to confer the highest activity in other 1,2-diamino-cyclohexane platinum (II) complexes (see, for example, Gann 67: 921 - 922, 1976 and J. Clin. Hematol. Oncol. 8(2): 44 - 50, 1978). While a large number of platinum (II) complexes have been disclosed as having some antitumor activity, there remains a need for new platinum (II) complexes which will have more advantageous properties, i.e. greater activity, broader spectrum, less toxicity etc. Especially desirable are platinum complexes having significant advantages over cisplatin, the only platinum complex presently being marketed as an anticancer agent. The present invention provides novel platinum (II) complexes of the formula wherein the stereoisomerism of 1,2-diaminocyclohexane is cis(-), trans(+)- or preferably, trans(-)- and R¹ and R² when taken together represent a group of the formula and of the formula: The complexes provided according to the present invention demonstrate high antitumor activity when tested against experimental mouse tumors. The complexes of the present invention are prepared from the corresponding dichloro platinum (II) complexes which have been described in the literature. Synthesis of the dichloro complexes of 1-aminomethylcyclooctylamine and 1,2-diamino-2,4-dimethylpentane is disclosed, for example, in J. Inorg. Biochem. 11: 139 - 149 (1979) while synthesis of cis-, trans(+)- and trans(-)-dichloro(1,2-diaminocyclohexane)platinum (II) is described, for example, in U.S. patent 4,169,846. As mentioned above, while any of the cis-, trans(+)- or trans(-)-isomers of the dichloro(1,2-diaminocyclohexane)platinum (II), or the mixture of isomers, can be used as the starting material to prepare the novel complexes of 1,2-diaminocyclohexane disclosed herein, it is preferred to use the trans(-)-starting material since the biological activity of the end-products is somewhat higher using this material. Synthesis of the complexes of the present invention using the dichloro platinum (II) complexes is carried out by conventional procedures used to make other platinum (II) complexes known in the art. In one suitable procedure, for example, the appropriate dichloro complex is reacted with the silver salt of the desired ligand. Typically about 1 mmol of the dichloro complex is reacted with about 0,95 mmol of the appropriate disilver salt in water. AgCl precipitates from solution and is filtered off. The desired complex is then isolated from the filtrate as by evaporation at low temperature in vacuo. The product, if desired, may be washed with an appropriate organic solvent (in which it is substantially insoluble) prior to drying in vacuo. This procedure is used, for example, in Examples 6, 8 and 9 below. Another suitable procedure involves first forming in situ the dinitrato platinum (II) complex and then reacting this intermediate with an alkali metal salt of the desired ligand. Typically about 1 mmol of the dichloro complex is reacted with about 1.95 mmol AgNO₃ in water in the dark at 20 - 70°C for 1 - 3 hours. The reaction mixture is then generally stirred at room temperature for about 4 hours, chilled (e.g. to about 4°C) and filtered. To the filtrate is added an aqueous solution of about 1 mmol (in the case of a bidentate ligand) or 2 mmol (in the case of a monodentate ligand) of the desired ligand. If sufficiently insoluble in water, the desired complex generally precipitates on heating to about 70 - 80°C and gradual cooling to 4°C. The precipitated complex can then be recovered by filtration. This procedure is appropriate if the desired complex is of low aqueous solubility and is illustrated in Example 3 below. An especially preferred procedure involves conversion of the dichloro starting material to the dihydroxy complex and then reaction of this intermediate with the desired ligand. Typically an aqueous solution of the dichloro complex is reacted with Ag₂SO₄ and stirred in the dark at room temperature for about 24 hours. The resulting solution is filtered. The absence of silver ions in the filtrate indicates the reaction is complete and that the sulfato complex has been formed in solution. There is then added to the filtrate Ba(OH)₂ and the solution is stirred in the dark for another 24 hours. The solution is then filtered to remove BaSO₄. At this point the aqueous filtrate containing the dihydroxy platinum (II) intermediate in solution is then reacted with the desired ligand to give the corresponding end-product which can be isolated from aqueous solution, for example by evaporation. This procedure is illustrated in Examples 1 - 2, 4 - 5, 7 and 10 - 14 below. Product complexes obtained according to the present invention generally require no further purification. However, if desired, the complexes may be purified by conventional procedures, for example by dissolving in a suitable solvent (e.g. water, diemthylformamide, ethanol etc.), and precipitating from solution by addition of a large excess of a solvent in which the desired complex is only slightly soluble (e.g. ethanol, acetone, diethyl ether, etc.). Products can then be dried over anhydrous silica gel in vacuo in the dark. Complexes may be tested for identity and homogeneity by IR spectroscopy and thin layer chromatography using the TLC systems described by Hall, et al in J. Inorg. Biochem. 11: 139 - 149 (1979). Biological Properties of Complexes The antitumor activities of the complexes provided by the present invention have been determined against L-1210 leukemia in the mouse and, for certain of the complexes, against other mouse tumor systems. Results of antitumor testing against the transplantable mouse tumors L1210 leukemia, cisplatin-resistant L-1210 leukemia, B16 melanoma, Madison 109 lung carcinoma and colon 26 carcinoma are shown below in Tables 1 and 2. In Table 1 are shown the maximum % T/C values obtained for each platinum compound versus L-1210 leukemia using both single and 9 d 1 → 5 dosing. Also shown in Table 1 are testing results against a cisplatin resistant line of L-1210 leukemia. Table 2 shows the maximum % T/C values obtained for each of the selected compounds against ip B16 melanoma, C26 carcinoma and M109 tumors. Also shown are the concomitant maximum effects obtained for cisplatin. A ratio of each compounds maximum effect relative to that of cisplatin is also included. The results to date from side-effects (toxicity) testing of compounds of the present invention are summarized in Table 3 with more detailed data shown in Tables 4 and 5. LD₅₀ values (single does ip) were determined in BDF₁ mice for certain complexes, i.e. those of Example 3 and 4. The other compounds were in such short supply that they were tested for effects on BUN values using the optimum dose (O.D.) from the L-1210 test for dose selection. Results of emesis testing are shown in Table 5 for four compounds. The method used to evaluate the platinum complexes for antitumor activity and toxicity in animals are summarized below. These methods have been published elsewhere in more detail (Rose, et al, Cancer Treat. Rep. 66, 135, 1982; Florczyk, et al, Cancer Treat. Rep. 66: 187, 1932). Animals Male and female BDF₁, CDF₁, DBA/2, BALB/c and C57BL/6 mice (17 - 21 gm) were used for the antitumor evaluations. The toxicity studies were done in male BDF₁ mice (25 - 29 gm) and castrated male Fitch ferrets (1 - 1,5 kg). Tumors The parent line of L1210 leukemia (L-1210) was maintained in DBA/2 mice by ip transplantation of 10⁵ cells at weekly intervals. A subline of L-1210 resistant to cisplatin (L-1210/CDDP) was maintained in the same manner but the passage mice were treated ip with cisplatin, 4 mg/kg, on Day 4 post-implant. B16 melanoma (B16) was maintained by sc transplant in C57BL/6 mice every two weeks. Madison 109 lung carcinoma (M109) and colon 26 carcinoma (C26) were maintained by sc transplant in BALB/c mice every two weeks. Antitumor Testing The level and route of tumor inoculum, host, group size, and drug treatment schedule and route for each tumor type are summarized below in Table 6. Each complex was evaluated initially against L-1210 at 4 dose levels per schedule selected on the basis of lethality data provided by the Wadley Institute. In subsequent experiments with the other tumors the complexes were evaluated at a minimum of 3 dose levels selected on the basis of maximum tolerated levels in the L-1210 experiments. Cisplatin-treated groups and a saline-treated (or untreated) tumor control group were included in each experiment. The L-1210 experiments were terminated on Day 30 whereas the B16, C26 and M109 experiments were terminated on Day 60 (some C26 experiments were terminated on Day 75). Mice alive at the termination day were autopsied and considered cured if no tumor was visible. Mice were observed daily and comparative antitumor activity relative to cisplatin was determined based on the fraction of mice cured and % T/C, defined as: Median survival time of treated mice Median survival time of untreated mice x 100 A complex was considered active if it produced a % T/C value ≧ 125. Toxicity Testing Single-dose ip LD₅₀ values were determind in BDF₁ mice for the indicated complexes. The mice, 5-10/dose, were observed for 30 days after dosing and the LD₅₀ value calculated by the Weil method (Biometrics 8: 249, 1952). The complexes that were active against L-1210 were evaluated for nephrotoxicity by determining their effects on blood urea nitrogen (BUN) levels in mice. Groups of 40 - 50 mice/experiment were bled from the retro-orbital plexus and individual BUN values determined by a modified urease method using a Centrifichem System 400. The test complex, or saline for controls, was given at several dose levels (5-10 mice/dose) in single ip injections. The highest dose of the complex usually corresponded to its LD₅₀ value or 1,5 times its optimal single dose in the L-1210 experiment. At least two lower doses were included to evaluate dose-related effects. The BUN values were measured 4 and 7 days after dosing. BUN values ≧ 30 mg% were considered indicative of drug-induced nephrotoxicity. Selected complexes were evaluated for emetic effects in ferrets. The test drug was administered iv as a bolus and the animals observed for 6 hours. The time to the first emetic episode (retching and expulsion of vomitus) and the number of episodes were recorded. The initial test dose of each complex was selected based on its potency, relative to cisplatin, to cause lethality in mice. Subsequent test doses were selected based on the emetic response and lethality of the initial dose. As illustrated by the data shown above, the platinum compounds of the present invention exhibit inhibitory activity against malignant tumors in mammals. The compounds of the invention are suitable for therapeutically treating a mammalian host affected by a malignant tumor by administering to said host an effective tumor-inhibiting dose of a compound of the present invention, i.e. the products described in the Examples. According to another aspect, a pharmaceutical composition is provided which comprises a tumor-inhibiting amount of a compound of the present invention in combination with a pharmaceutically acceptable carrier or diluent. The platinum compounds of the present invention are preferably administered parenterally in the same manner as other antitumor platinum complexes known in the art, e.g. cisplatin. Preparations for parenteral administration include sterile aqueous or non-aqueous solutions, suspensions or emulsions. They may also be manufactured in the form of sterile solid compositions which can be dissolved in sterile water, physiological saline or some other sterile injectable medium prior to administration. It will be appreciated that the actual preferred dosage amounts used will vary according to the particular compound being used, the particular composition formulated, the mode of application and the particular situs, host and disease being treated. In general the compounds are injected intraperitoneally, intravenously, subcutaneously or locally. Many factors that modify the action of the drug will be taken into account by those skilled in the art, for example, age, body weight, sex, diet, time of administration, route of administration, rate of excretion, condition of the host, drug combinations, reaction sensivities and severity of the disease. Administration can be carried out contineously or periodically within the maximum tolerated dose. Optimal application rates for a given set of conditions can be readily ascertained by those skilled in the art using conventional dosage determination tests. Illustrative methods for preparing the compounds of the present invention are provided below. These examples, however, are not meant to limit the scope of the invention to the specific procedures employed. Volume ratios used to describe solvent systems are volume/volume. Example 1 Preparation of thymidylato trans(-)-1,2-diaminocyclohexane platinum (II) A mixture of dichloro (1,2-diaminocyclohexane) platinum (II) (760 mg) and 1,08 g of disilver thymidine monophosphate was suspended in 30 ml of water. The reaction mixture was stirred in the dark at room temperature for 36 hours, silver chloride was filtered off and the reaction mixture and then evaporated to dryness. The title product obtained (698 mg)was a white material that is water soluble but alcohol-insoluble. When tested in the TLC systems indicated below, the following R f values were obtained: The IR was consistent for the indicated structure. Example 2 Preparation of phosphonoacetato trans(-)-1,2-diaminocyclohexane platinum (II) A mixture of cis-dihydroxy-1,2-diaminocyclohexane platinum (II) (646 mg) and phosphonoacetic acid (280 mg) in 15 ml of water was heated at 50°C for 1 hour. The reaction mixture was evaporated to dryness and washed with ethanol to remove any unreacted starting materials. The product obtained (510 mg) is water-soluble but alcohol-insoluble. IR indicated that phosphonoacetic acted as a bidentate ligand. When tested in the TLC systems indicated below, the following R f values were obtained: Example 3 Preparation of hydroxymethylphosphonato trans(-)-1,2-diaminocyclohexane platinum (II) A mixture of cis-dihydroxy-1,2-diaminocyclohexane platinum (II) (686 mg) and hydroxymethylphosphonic acid (224 mg) in 15 ml of water was heated for 1 hour at 50°C. The resulting solution was evaporated to dryness to give title product. The product is soluble in both water and methanol. The product was dissolved in water and precipitated with acetone to remove unreacted starting materials. Yield: 358 mg. When tested in the TLC systems indicated below, the following R f values were obtained: The IR was consistent for the indicated structure. Example 4 Preparation of phosphonoacetato 1-aminomethylcyclooctylamine platinum (II) A mixture of cis-dichloro-1-aminomethylcyclooctylamine platinum (II) (1,266 g) and 936 mg Ag₂SO₄ was suspended in 30 ml water and stirred in the dark at room temperature for 24 hours (the filtrate tested negative for Ag⁺) Ba(OH)₂ 8H₂O (945 mg) was added to the filtrate and the resulting solution was stirred in the dark for 24 hours. The solution was filtered to remove BaSO₄. The cis-dihydroxy-1-aminomethylcyclooctylamine platinum (II) derivative was not isolated. Phosphonoacetic acid (430 mg) was added to the filtrate and the solution was heated for 1 hour at 50°C. The color of the filtrate changed from yellow to colorless. Upon evaporation of the solution to dryness and washing of the resulting solid with methanol, there was obtained 0,70 g of title product as a light-greenish-white solid. The product is very water-soluble. When tested in the TLC systems indicated below, the following R f values were obtained: The IR was consistent for the indicated structure.

Claims: Claims for the following Contracting States : BE, CH, DE, FR, GB, IT, LI, LU, NL, SE 1. A compound of the formula wherein the stereoisomerism of 1,2-diaminocyclohexane is cis-, trans(+) or trans(-) and R¹ and R² when taken together represent a group of the formulae 2. A compound of claim 1 wherein the stereoisomerism of 1,2-diaminocyclohexane is trans(-). 3. The compound having the formula 4. A process for preparing the compounds of claims 1 to 3, characterized in that the appropriate dichloro platinum (II) complex is reacted with the silver salt of the desired ligand R¹ and R², or the dichloro platinum (II) complex is converted to the dinitrato platinum (II) complex which is then reacted with an alkali metal salt of the desired ligand R¹ and R², or the dichloro platinum (II) complex is converted to the dihydroxy complex which is then reacted with the desired ligand. 5. A pharamaceutical composition comprising at least one compound of claims 1 - 3 in combination with a pharmaceutically acceptable carrier or diluent. Claims for the following Contracting State : AT 1. A process for preparing a compound of the formula I wherein the stereoisomerism of 1,2-diaminocyclohexane is cis-, trans(+) or trans(-) and R¹ and R² when taken together represent a group of the formulae or the compound having the formula characterized in that the appropriate dichloro platinum (II) complex is reacted with the silver salt of the desired ligand R¹ and R², or the dichloro platinum (II) complex is converted to the dinitrato platinum (II) complex which is then reacted with an alkali metal salt of the desired ligand R¹ and R², or the dichloro platinum (II) complex is converted to the dihydroxy complex which is then reacted with the desired ligand. 2. The process of claim 1 wherein the stereoisomerism of the 1,2-diaminocyclohexane radical in the compounds of formula I is trans(-).

Inventors: SPEER, ROBERT JAMES, STEWART, DAVID PHILIP
Applicants: BRISTOL-MYERS SQUIBB COMPANY
Application year: 1985
Main classification: C07F15
IPC codes: C07F15, A61P35, A61K31, C07F9, C07C211, C07H23