JPS6254430B2 - - Google Patents

Description

[Detailed description of the invention]

The present invention relates to a coprecipitate obtained from a neotissue formation-inhibiting metal complex compound and a hydrophilic polymer, a method for producing the coprecipitate, and a sterile aqueous solution containing the coprecipitate. Recently, a drug containing a cis-diaminedichloroplatinum () complex compound has been commercially available as an anticancer chemotherapeutic agent. This compound, called Cisplatin, has been found to be a very effective anticancer agent, especially in the treatment of testicular tumors, but also, for example, ovarian tumors and autocellular carcinoma. The disadvantage of cisplatin is its relatively high toxicity. In particular, its effects leading to nephrotoxicity and persistent hearing loss are particularly serious. Kidney damage and hearing damage were already observed with a significant frequency after administration of only one therapeutic dose. In addition to the nephrotoxic and hematotoxic effects, the persistent severe nausea and associated nausea are extremely unpleasant for the patient. Recently, a large number of platinum complex compounds (West German Patent Application No. 2445418, West German Patent Application No. 2837237, West German Patent Application No. 2626559) have been developed.
No. 2,539,179) and other transition metal complexes have been proposed as cytostatic agents. West German Patent Application No. 2801355 describes a brown amorphous complex compound obtained by the reaction of ascorbic acid, a titanium () compound and a copper () compound (molar ratio 36:1:6). It has been described as having curative and prophylactic effects against leukemia. It has been reported that titanocene, zirconocene and hafnocene dichloride have an inhibitory effect on Ehritz's condensate tumor in mice. [P. Ko¨pf-Maier, B. Hess, H.
Ko¨pf: J. Cancer Res. Clin. Oncol.
96 , 43 (1980)]. Dialkyl-tin dihalogen. A phenanthrin- or -bipyridyl-complex compound was described to have an inhibitory effect on P388 leukemia in mice [AJ Crowe and PJ Smith: Chemistry and Industry;
1.Ma¨rz 1980, P.200] By the way, the general formula: [R ¹ (CH ₂ ) _n C(O)CR ³ C(O)R ² ] _2+o MX _2-o
() [In the formula, M represents titanium, zirconium or hafnium, and R ¹ is 4-chromium, 4-brome, 3
-nitro, 4-methoxy, 3,4-dimethoxy,
Represents a phenyl group optionally substituted with 4-methyl or 2,4,6-trimethyl, R ² represents a methyl group or R ¹ , R ³ represents hydrogen or chlorine, and X represents fluorine, chlorine or represents bromine, but not fluorine when M represents zirconium or hafnium, m is 1 or 0, and n is 0 or 1 or 0 when M represents zirconium or hafnium]. It was found that the complex compound shown has a cytostatic effect comparable to, but much less toxic than, cisplatin. Medicaments containing one or more of the compounds are suitable for cancer disease control. In particular, dichlorobis(benzoylacetonato)
Preference is given to agents containing titanium (), dichlorobis(benzoylacetonato)zirconium () and/or dichlorobis(benzoylacetonato)hafnium (). The drug is administered intravenously, but also intramuscularly, intraperitoneally, subcutaneously, rectally or orally. External applications are also possible. Administration by intravenous injection or infusion is preferred. If the pharmaceutical formulation containing the complex compound contains pharmaceutical excipients in addition to the complex compound (active substance), the active substance content of this formulation is equal to or greater than that of the total formulation.
0.1-99.5% by weight, preferably 0.5-95% by weight. The active substance may be applied in any suitable form, provided that its sufficient content is ensured and maintained. For example, it can be achieved by oral or parenteral administration of appropriate doses. Advantageously, the pharmaceutical formulation is in the form of unit doses adapted to the desired administration. The unit dose may be, for example, a fixed volume of a tablet, dragee, capsule, suppository or powder, particle, solution, emulsion or suspension. "Unit dose" as used above means a physiologically specific unit containing a single amount of the active ingredient together with pharmaceutical excipients, where the active substance content in the unit dose is a fraction of a single therapeutic dose. Or double the amount. A single dose is preferably administered at the time of application. Contains an amount of active substance that usually corresponds to all, half, one-third or one-quarter of the daily dose. Pharmaceutical formulations are present in unit doses, e.g. when specified for human application, about 0.1
~500mg, advantageously 10-200mg and especially 50-150mg
Contains active substances. Generally, in human medicine, in order to achieve the desired effect when administered parenterally, the active substance should be administered in daily doses of about 0.1 to about 5 mg/kg body weight, preferably 1 to 3 mg/kg body weight; Several single doses, advantageously 1~
It has been found advantageous to administer it in the form of three single doses. A single dose contains the active substance from approximately 0.1 to kg body weight
It is contained in an amount of about 5 mg, preferably 1 to 3 mg. Therapeutic administration of the pharmaceutical formulation is carried out at fixed or variable times 1 to 4 times a day, e.g. after meals and/or
Or you can do it in the evening each time. However, deviations from these doses may be necessary depending, inter alia, on the nature of the patient being treated, his weight and age, the nature and severity of the disease, the method of preparation and application of the drug and the period of time during which the administration takes place. Sometimes. Thus, in some cases it may be sufficient to make do with less than the above-mentioned amount of active substance, while in other cases the above-mentioned amount of active substance must be exceeded. In order to reduce the risk of side effects, treatment with the agents according to the invention may also be combined with the administration of other cytostatic agents with different levels of action, as is customary in the case of medical tumor therapy. good. It may also be advantageous to carry out treatment with periodic cytostatic therapy. In this case, each treatment is followed by a recovery period. This utilizes the experience that healthy tissue in most organs regenerates more rapidly than malignant tissue. The optimal dose of the active substance required in each case and the method of application can be readily determined by a person skilled in the art on the basis of his or her common knowledge. Pharmaceutical preparations generally consist of a complex compound and a pharmaceutically acceptable non-toxic medicinal excipient which is used in solid, semi-solid or liquid form as an additive or diluent for the therapeutically active ingredient. or as a coating, for example in the form of capsules, tablet shells, bags or other containers. Excipients can also be used, for example, as enhancers of drug absorption by the body, as formulation aids, sweeteners, flavoring agents, colorants or preservatives. For parenteral application of the drug, dispersants or wetting agents and/or pharmacologically acceptable diluents such as propylene or butylene glycol, and/or solubilizing agents such as Tween, glycerin-polyethylene glycol ricinoleate [cremophor ( Cremo phore EL)] or sterile injectable aqueous dispersions, isotonic salt solutions, or other solutions containing polyvinylpyrrolidone are used. In particular, after mixing a solution of the complex compound in a water-free organic solvent with a solution of a hydrophilic polymer, such as polyvinylpyrrolidone (PVP) or polyoxyethylene sorbitan fatty acid ester (trade name Tween) and removing the solvent. Advantageously, the remaining residue is administered as an aqueous solution. Suitable organic solvents include, for example, chloroform or methylene chloride, which is made anhydrous in the usual manner before use. The hydrophilic polymer is 5 to 30 times larger than the complex compound, preferably 10 to 30 times larger than the complex compound.
It has been found advantageous to use a 20 times excess weight. It is also possible to introduce the polymer itself into the solution of the complex compound. The residue remaining after removal of the solvent is preferably freed of solvent residues as thoroughly as possible under high vacuum. Depending on the type and amount of hydrophilic polymer used, solid crystalline or glassy residues or liquid or sticky residues are obtained. The latter residue is generally converted by cooling into a solid, often waxy product. These residues should be referred to as coprecipitates. Aqueous solutions are prepared by treating the coprecipitate with water. PVP coprecipitates generally dissolve already at room temperature. For the coprecipitate with polyoxyethylene sorbitan fatty acid ester, the coprecipitate and water are heated at 25-60℃, preferably 30-40℃, before combining.
It can advantageously be dissolved by heating to .degree. The complexes according to the invention can be prepared by reacting metal tetrahalides with the corresponding diketones in a manner known per se in an inert solvent such as benzene, n-hexane, diethyl ether, methylene chloride or chloroform with careful removal of water. Manufactured by [Zirkon-und Hafnium complexe (Zirkon-und Hafnium complex)
Hafniumkomplexe): M.Cox, J.Lewis and RS
Nyholm, J. Chem. Sos. 1964 , 6113; Zinnkomplexe: W. Dilthey, J.
Prakt. Chemie [2] 111 , 147 (1925); W.
Dilthey, Ber.Deutsch.Chem.Ges. 36 , 923
(1903); GTMorgan, HDKDrew, J.Chem.
Soc. 1924 , 373; WH Nelson, Inorgan.Chem.
6 , 1509 (1967); RWJones, Jr.uRCFay,
Inorgan.Chem. 12 , 2599 (1973); Titanium complexe: N. Serpone and RC
Fay, Inorgan. Chem. 6 , 1835 (1967)]. The reaction is advantageously carried out in a dry protective gas atmosphere, for example nitrogen. The solid metal tetrahalide is present as a suspension in the same solvent in which the diketone is dissolved, and the diketone solution is added dropwise thereto.
In the reaction of liquid metal tetrahalides, the diketone is preferably dissolved in an inert solvent and the metal tetrahalide is added dropwise. It has been found advantageous to add the diketone in up to a 20% excess, preferably about 10% excess, over the theoretically required amount. Depending on its strength, the reaction is carried out at room temperature, for example with cooling or heating under reflux. It may be necessary to heat the reaction mixture under reflux for 1 to 3 days to complete the reaction. It has proven advantageous to drive off the hydrogen halide formed during the reaction by passing dry nitrogen through the reaction mixture.
After the reaction is complete, unreacted solid residues of the tetrahalide or undissolved hydrolysis products are optionally removed, and the reaction solution is finally concentrated or cooled to precipitate the product. 1,3-diketones are known and can be produced by methods known per se. For example, the same compounds can be prepared by ester condensation of the corresponding arylmethylkene and ethyl acetate or of the corresponding arylacetic ethyl ester and sodium amide as condensing agent (JT
Adams, CRHauser.J.Amer.Chem.Soc. 66 ,
1220 [1944]). 1,3-diketones can also be obtained by adding the corresponding arylmethylketone dissolved in acetic acid ethyl ester to a suspension of sodium in benzene or toluene (DWBrown, SFDyke, M.Sainsbury, G.
Hardy, J. Chem. Soc. (c) 1971 , 3219). Also,
Arylmethyl ketones and acetic anhydride can also be reacted in the presence of boron trifluoride. [HG
Walker, CR Hauser, J. Amer. Chem. Soc. 68 ,
2742 (1946)]. 2-Chlor-1,3-diketones are prepared by chlorination of the corresponding 1,3-diketones with sulfuryl chloride in ether [Macbeth, J.Chem.Soc. 123 , I, 1129]
(1923)]. 1-benzyl-1,3-diketone is
It is prepared by condensing the corresponding phenylacetic acid ethyl ester with the corresponding methyl ketone in the presence of sodium amide [A. Becker, Helv.
Chin, Acta 149 , 1114 (1949)]. Next, the present invention will be explained by reference examples and examples, but these examples do not limit the present invention. "Ether" means diethyl ether. Production example (reference example) 1 Dibromobis(benzoylacetonato)zirconium () Zirconium tetrabromide 10 in 200 ml of dry ether
to a suspension of 100 g of dry ether under a stream of dry nitrogen.
24 g of benzoylacetone dissolved in ml are added dropwise, the reaction mixture boiling under reflux. The reaction mixture is heated under reflux for 24 hours. The precipitate that forms is washed twice with 100 ml each of ether and dissolved in about 250 ml of chloroform. Concentrate the solution until it begins to become cloudy. A colorless crystalline sludge precipitates on cooling to room temperature. Melting point 220-221 after three times recrystallization
1 g of the title compound (theoretical value) at °C (red melt)
8.5%). 2 Bromotris(benzoylacetonato)zirconium () Zirconium tetrabromide 11 in 200 ml of dry benzene
13 g of benzoylacetone dissolved in 100 ml of dry benzene are added dropwise under a stream of nitrogen and at boiling reflux. The reaction mixture is boiled under reflux until the evolution of hydrogen bromide has ceased. Pale brown crystals precipitate from the reddish-brown solution that forms upon cooling. This crystal,
Treat with ether under reflux until the ether no longer exhibits a yellowish color. After drying the crystals in an oil pump vacuum, 10 g (57% of theory) of the title compound with a melting point of 156° C. are obtained. 3 Dichlorobis(benzoylacetonato)hafnium () 5 g hafnium tetrachloride in 200 ml dry ether
15.2 g of benzoylacetone dissolved in 100 ml of dry ether are added dropwise to the suspension at reflux boiling under a stream of nitrogen. After boiling under reflux for 24 hours, the precipitate formed was collected and washed twice with 100 ml of ether each time.
Dissolve in approximately 300 ml of chloroform. Concentrate until cloudy, filter, and leave the solution at -5°C. The colorless precipitate is recrystallized from chloroform. 3 g (33.7% of theory) of the title compound are obtained, melting point 231 DEG -232 DEG C. (red melt). 4 Dibromobis(benzoylacetonato)hafnium () 2 g hafnium tetrabromide in 100 ml dry ether
2 g of benzoylacetone dissolved in 100 ml of dry ether are added to the suspension under reflux under a stream of nitrogen. After the evolution of hydrogen bromide has ended, the red precipitate is removed and extracted five times with ether under reflux for 2 hours each time. 1 g (25% of theory) of the title compound is obtained. 5 Dibromobis(benzoylacetonato)titanium () Titanium tetrabromide dissolved in 40 ml of dry benzene
Add 8.0 g (0.050 mol) of benzoylacetone, 7.0 g (0.019 mol) dissolved in 100 ml of dry benzene. The reaction mixture is heated under reflux for 30 minutes, a stream of dry nitrogen being passed through the solution. After distilling off about 40 ml of solvent, the brown-red product is collected, recrystallized twice from a benzene/hexane mixture and dried at 80° C. under reduced pressure. Melting point 210-222℃. 6 Difluorobis(benzoylacetonato)titanium () 17.4 g (0.108 mol) of benzoylacetone dissolved in 85 ml of methylene chloride was gradually dissolved with stirring into 5.40 g of titanium tetrabromide in about 350 ml of methylene chloride.
(0.0436mol) into a suspension. A slow flow of dry nitrogen is passed through the reaction mixture until the evolution of hydrogen fluoride has ceased. This lasts about 20 hours. The filtered yellow-orange solution is then concentrated under reduced pressure to approximately half the volume. On cooling, a yellow product crystallizes out, which is filtered off and dried under vacuum (yield 11.0 g, corresponding to 62% of the theoretically calculated amount). Melting point: 196~
198.5℃. 7 Dichlorobis(benzoylacetonato)titanium () 2 ml (0.018 mol) of freshly distilled titanium tetrachloride are dissolved in 0.8 g (0.05 mol) solution. During this time the reaction mixture is heated and a dense fog of hydrogen chloride escapes. At the same time, the reaction mixture gradually acquires a deep red color.
Once the reaction has slowed down, the solution is boiled at reflux until all hydrogen chloride gas is removed (approximately 2.5 hours). Orange crystals precipitate on cooling. After decanting the upper layer, the crystals are washed with benzene and dried under reduced pressure at room temperature. Yield: 11.68 g of yellow-orange crystals. Melting point: 209
~210℃ (decomposition). Identification can be done spectroscopically, e.g.
This is done by NMR. Furthermore, the following table shows other preparation examples with analytical values and melting points:

【table】

[Table] Example Co-precipitation of dichlorobis(benzoylacetonato)titanium () 100 mg of dichlorobis(benzoylacetonato)titanium () is dissolved in about 10 ml of anhydrous methylene chloride. To this clear solution is added a solution of 1 g of PVP30BT or 2 g of Tween 60 in approximately 30 ml of anhydrous methylene chloride. Stir the resulting solution for several minutes. It is also possible to dissolve an appropriate complex compound in a large amount of solvent and introduce the polymer as it is into the solution.
The solvent is removed in a rotary evaporator. Solvent residues are removed by applying high vacuum. When using PVP30BT, a red-yellow crystalline material is obtained, which is about 8
Soluble in water to mg/ml. When using Tween, a red sticky substance is obtained. To produce an aqueous solution, the coprecipitate and water should be combined for 30 to 40 minutes before combining.
It is advantageous to heat to .degree. The coprecipitate dissolves to about 17 mg/ml of water. Pharmacological effects of complex compounds (reference) A Tumor type 1 Sarcoma 180 tumor type Female NMRI mice, approximately 6 weeks old and weighing 18 to 20 g, were given approximately sarcoma 180 tumor cells in 0.2 ml of physiological saline solution.
16 ⁶ each inoculated intraperitoneally (ip). The same tumor is passed in mice of the same strain. Tumor cells are harvested from freshly killed mice immediately before transplantation. Mice are randomly selected for inoculation. Six mice are used per dose. Cisplatin and cyclophosamide are used as control substances. The number of control groups (untreated animals) is selected to correspond to the square root of the total number of groups. The substance is injected intraperitoneally 24 hours after implantation, in each case as a suspension in a customary solubilizer, such as Tween (a polyoxyethylene derivative of the sorbitan ester). 2 L1210 Leukemia Tumor Type in NMRI Mice Female NMRI mice, approximately 6 weeks old and weighing 18-20 g, are each inoculated intraperitoneally with approximately ¹⁶⁶ freshly harvested tumor cells in 0.2 ml of physiological saline solution. The tumor is passed in mice of the same strain. The number of animals, number of control groups and administration of the substance are the same as those of the Sarcoma 180 tumor model described in 1. 3 Walker 256 carcinoma type Female SD rats weighing approximately 150 g are each inoculated intraperitoneally with approximately 10 ⁶ tumor cells in 0.5 ml of physiological saline solution. The same tumor is passed through rats of the same strain.
The number of animals and administration of the substance are the same as for the sarcoma 180 tumor type described under 1. Six animals are used per dose. B. Experimental Results The following table summarizes the results obtained from the Sarcoma 180 tumor model described in A.1. The stated dose was applied once at the beginning of the experiment as described above. The factor T/C, expressed in %, represents the percentage increase in the median survival time of treated animals relative to the median survival time of untreated control animals. In part of the table, the cure rate, ie the number of cured animals relative to the number of treated animals, is listed in parentheses after the factor T/C. The experiment is stopped as soon as the median survival time T/C of treated animals reaches 300% of the median survival time of untreated animals. To calculate the median survival time, animals that are still alive at the end of the experiment are taken over those that are dead at the end of the experiment. The comparison compound cisplatin is therapeutically effective at a dose range of about 8-10 mg/Kg, and the animals are mostly cured. The median survival time of animals treated with cisplatin is already shorter than the same time of untreated control animals at a dose of approximately 20 mg/Kg.

【table】

【table】

[Table] Figure 1 shows the compounds cyfluorobis(benzoylacetonato)titanium ( ) [curve a], dichlorobis(benzoylacetonato)titanium ( ) [curve b] and dibromobis(benzoylacetonato)titanium [curve b] with respect to the L1210 leukemia tumor type. These are the results of a comparative experiment using curve c]. The median survival time (days) is plotted on the ordinate and the single dose administered at the beginning of the experiment is plotted on the abscissa. The median survival time (18 days) of untreated control animals is marked as a horizontal line (d). Furthermore, the cure rate (number of cured animals per group of 6 animals) is also given in parentheses for the individual doses. [For example (0/6), (1/6),
(2/6) etc.] The experiment was stopped after 60 days. As a comparative substance, cisplatin was administered at a dose of 8 mg/Kg. Figure 2 shows the compounds difluorobis(benzoylacetonato)titanium (curve a), dichlorobis(benzoylacetonato)titanium (curve b) and dibromobis(benzoylacetonato)titanium (curve b) for the Walker 256 carcinosarcoma tumor type. The results of a comparative experiment using ( ) [curve c] are shown.
The median survival time (days) is plotted on the ordinate and the single dose administered at the beginning of the experiment is plotted on the abscissa. The median survival time of untreated control animals (4.5 days) is marked as a horizontal line. These experiments show that the test complexes are significantly more effective than cisplatin. At the same time, the toxicity of these compounds is much lower than that of cisplatin (LD ₅₀ of 12 mg/Kg in mice and rats). The nephrotoxicity known to exist in the case of cisplatin could not be observed in the case of the test complex. Pharmacological effects of coprecipitate.
5×10 ⁶ sarcoma 180 cells were implanted subcutaneously. Six of the same animals were treated with Tween in an aqueous solution using dichlorobis(benzoylacetonato)titanium () 4 mg/Kg twice a week for 4 weeks starting from the second week after transplantation.
Treat intravenously as a 60 coprecipitate (1:20). The treated animals were cured after treatment, whereas the untreated animals had sarcomas about the size of walnuts, most of which were already centrally gangrene and ulcerated in several areas. In another experiment, zero-day-old female SD rats were given
5×10 ⁶ Yoshida sarcoma cells were subcutaneously transplanted. Dichlorobis(benzoylacetonato)titanium () 4mg/Kg twice in the second week Tween 60 in aqueous solution
(trade name) was administered intravenously as a coprecipitate (1:20). The tumor regressed. After a 3-week break, two to three animals developed recurrent tumors the size of snails. One was cured. Recurrent tumors were treated intravenously twice weekly at the original dose for 4 weeks. Two animals were cured. Control animals showed enlarged tumor growth at this point and died after a short period of time.

[Brief explanation of the drawing]

FIG. 1 is a graph showing the results of comparative experiments using compounds according to the invention on the L1210 leukemia tumor type, and FIG. 2 is a graph showing the results of said experiments on the Walker 256 carcinosarcoma tumor type. a...Difluorobis (benzoylacetonate)
When using titanium (). b...When using dichlorobis(benzoylacetonato)titanium (). c...When using dibromobis(benzoylacetonato)titanium (). d...Untreated control animal.

Claims (1)

[Claims] 1 General formula: [R ¹ (CH ₂ ) _n C(O)CR ³ C(O)R ² ] _2+o MX _2-o
() [In the formula, M represents titanium, zirconium or hafnium, and R ¹ is 4-chromium, 4-brome, 3
-nitro, 4-methoxy, 3,4-dimethoxy,
Represents a phenyl group optionally substituted with 4-methyl or 2,4,6-trimethyl, R ² represents a methyl group or R ¹ , R ³ represents hydrogen or chlorine, and X represents fluorine, chlorine or represents bromine but not fluorine when M represents zirconium or hafnium, m is 1 or 0 and n is 0 or 1 or 0 when M represents zirconium or hafnium A coprecipitate obtained from a complex compound represented by ] and a hydrophilic polymer. 2. The coprecipitate according to claim 1, wherein the hydrophilic polymer is polyvinylpyrrolidone. 3. The coprecipitate according to claim 1, wherein the hydrophilic polymer is polyoxyethylene sorbitan monofatty acid ester (Tween). 4 The hydrophilic polymer is polyoxyethylene (20)
The coprecipitate according to claim 3, which is sorbitan monostearate (Tween 60). 5. The coprecipitate according to claim 1, comprising dichlorobis(benzoylacetonato)titanium () and polyoxyethylene (20) sorbitan monostearate (Tween 60). 6 General formula: [R ¹ (CH ₂ ) _n C(O)CR ³ C(O)R ² ] _2+o MX _2-o
() [In the formula, M represents titanium, zirconium or hafnium, and R ¹ is 4-chromium, 4-brome, 3
-nitro, 4-methoxy, 3,4-dimethoxy,
Represents a phenyl group optionally substituted with 4-methyl or 2,4,6-trimethyl, R ² represents a methyl group or R ¹ , R ³ represents hydrogen or chlorine, and X represents fluorine, chlorine or represents bromine but not fluorine when M represents zirconium or hafnium, m is 1 or 0 and n is 0 or 1 or 0 when M represents zirconium or hafnium ] In producing the coprecipitate obtained from the complex compound and the hydrophilic polymer, the coprecipitation method is characterized in that a solution of the complex compound and the hydrophilic polymer dissolved in an inert organic solvent is concentrated and dried. How things are manufactured. 7 General formula: [R ¹ (CH ₂ ) _n C(O)CR ³ C(O)R ² ] _2+o MX _2-o
() [In the formula, M represents titanium, zirconium or hafnium, and R ¹ is 4-chromium, 4-brome, 3
-nitro, 4-methoxy, 3,4-dimethoxy,
Represents a phenyl group optionally substituted with 4-methyl or 2,4,6-trimethyl, R ² represents a methyl group or R ¹ , R ³ represents hydrogen or chlorine, and X represents fluorine, chlorine or represents bromine but not fluorine when M represents zirconium or hafnium, m is 1 or 0 and n is 0 or 1 or 0 when M represents zirconium or hafnium A sterile aqueous solution containing a coprecipitate obtained from a complex compound represented by ] and a hydrophilic polymer.