Complex platinum compounds, methods for their preparation and pharmaceutical composition

 

(57) Abstract:

Describes the new complex platinum compound of formula I, where X represents a halogen atom, is independently a hydroxyl group or a carboxylate group containing 1-6 carbon atoms, and a represents a group-NH2-R, where R is a tricyclic hydrocarbon fragment containing 10 to 14 carbon atoms, which may be optionally substituted in the tricyclic ring by one or two alkyl group(s), each containing 1-4 carbon atoms, a complex including the said platinum complex with beta - or gamma-cyclodextrin, which may be optionally substituted hydroxyalkyl groups containing 1-6 carbon atoms, a method of obtaining complex compounds of the formula I, based on the oxidation of the complex of bivalent platinum formula II with hydrogen peroxide and an optional substitution of hydroxyl groups in the resulting product carboxylate groups under the action of Alliluyeva agent, and a pharmaceutical composition. Disclosed complexes can be used as such or as part of a pharmaceutical composition for the treatment of cancer. 5 S. and 3 C.p. f-crystals, 12 PL.

Background of the invention

Complex compounds of platinum, effective as cytotoxic agents have been introduced into medical practice in the late seventies of this century. The first pharmaceutical product on the basis of complex compounds of platinum was cisplatin (CIS-diamin-dichloroplatinum (II)). During subsequent development of these complex compounds were synthesized and investigated dozens of platinum complex compounds; among them, carboplatin was the connection that received the highest value in Oncology. However, both the above-mentioned compounds should be administered only parenterally, and none of them are suitable for oral administration. In the publication J. Med. Chem. (1995), 38 (16), 3016-24 disclosed hexacoordination all-TRANS platinum complexes with halogen, hydroxyl or carboxylate groups and amino groups. The difference between these complexes and so on istochitsya to the complexes of platinum (IV), having a TRANS, TRANS, TRANS-geometry and having cyclohexylamino. The complexes CIS, CIS, TRANS actually getting under special conditions to a very limited extent only, for example, using a long acetylation carried out under the action of light irradiation. Otherwise, the complexes TRANS, TRANS, TRANS get in a normal acetylation. As for oral administration, in European patents EP 328274 and 423707 revealed some complex compounds of tetravalent platinum as suitable. These complexes of tetravalent platinum have CIS, TRANS, CIS-geometry and contain, in addition to four halogen or carboxylate ligands, two asymmetric groups, one of which is Eminem, and the second is a substituted alkyl - or cycloalkylation.

Currently still held the search of complex compounds of platinum, which will be higher antitumor efficacy compared with known complex compounds of platinum.

Now, in the framework of the present invention found some new complex compounds of platinum, which have a higher antitumor efficacy of ablaut the basis of the present invention.

Brief description of the invention

According to the first aspect of the invention provides a complex platinum compound of the formula (I):

< / BR>
where X represents a halogen atom,

In independently represent a halogen atom, a hydroxyl group or a carboxylate group containing 1-6 carbon atoms, and

A represents a group-NH2-R, where R is a tricyclic hydrocarbon fragment containing 10 to 14 carbon atoms, which may be optionally substituted in the tricyclic ring by one or two alkyl group(s), each containing 1-4 carbon atoms.

According to the second aspect of the invention provides a complex of the inclusion complex of platinum with oxidation state IV formula I

< / BR>
where X represents a halogen atom,

In independently represent a halogen atom, a hydroxyl group or a carboxylate group containing 1-6 carbon atoms, and

A represents a group-NH2-R, where R is a tricyclic hydrocarbon fragment containing 10 to 14 carbon atoms, which may be optionally substituted in the tricyclic ring by one or two alkyl group(s), each containing 1-4 carbon atoms,

with beta - or gamma-cyclodextrin, to the

Particularly advantageous complex platinum compounds of the present invention are complexes of the formula (I) where a represents adamantylamine, and X and have videopreteen values, and furthermore, their inclusion complexes with beta - or gamma-cyclodextrin, which may be optionally substituted as disclosed above.

Other predominant complex platinum compounds of the present invention are complexes of the formula (I), where a is 3,5-dimethyladamantane, and X and have videopreteen values, and furthermore, their inclusion complexes with beta - or gamma-cyclodextrin, which may be optionally substituted as disclosed above.

In another aspect the present invention provides a method of obtaining a complex platinum compounds of the formula (I), characterized in that the complex of bivalent platinum formula (II)

< / BR>
where X and a have the abovementioned meanings, are oxidized on the platinum atom hydrogen peroxide in the formation of dihydrochalcones platinum (IV), and optionally hydroxyl groups of the specified complex replaces the carboxylate groups under the action of Alliluyeva agent.

The invention is Stryn, which may be optionally substituted hydroxyalkyl group containing 1-6 carbon atoms, specified the method is characterized by mixing a solution of a platinum complex of the formula (I) in an organic solvent with an aqueous solution of beta - or gamma-cyclodextrin, which is optionally substituted hydroxyalkyl groups containing 1-6 carbon atoms, and at the next stage, the solvent is evaporated from the resulting solution.

Still another aspect of the invention is a platinum complex of the formula (I) above, or its inclusion complex with beta - or gamma-cyclodextrin for use as a pharmaceutical product.

The last aspect of the invention provides a pharmaceutical composition for the treatment of cancer, characterized in that it contains, as active compound, at least one platinum complex of the above formula (I) or its inclusion complex with beta - or gamma-cyclodextrin, which may be optionally substituted as specified above, and at least one pharmaceutical excipient.

Complex platinum compounds of the present invention are new chemical compounds, since of the properties have not been characterized, no way of obtaining them was not disclosed. The applicability of these compounds as active substances for the treatment of cancer is also new and inventive, because it was impossible to ascertain from the prior art, obvious way that the presence of the ligand primary tricyclic amine complexes of tetravalent platinum will lead to a significant enhancement of antitumor activity of new compounds of the present invention.

The main advantages of platinum complex compounds according to the present invention in comparison with hitherto known complexes of platinum and, namely, in comparison with the complex platinum compounds, disclosed in European patents EP 328274 and EP 423707 based not only on their higher efficiency by oral administration and low toxicity, namely on the broader spectrum of their antitumor activity.

Hereinafter the invention will be described in more detail by using examples of specific embodiments. You must understand that these examples are given for illustrative purposes and in no way limit the scope of the invention, which is defined by the claims.

Detailed OPI is Tina (IV) (hereinafter called "LA 12") [(OS-6-43)-bis(acetato) (1-adamantanamine)Emmendingen (IV)]

Mix 6.25 g (13.3 mmol) of b-(1-adamantylamine)-Amin-de-dichloro-AF-dihydroxylation (IV) [(OS-6-43)-(1-adamantanamine)immundiagnostica (IV)] at room temperature with an excess of acetic anhydride (50.2 ml, 532 mmol). During this from the original solution is continuously precipitated solid. After deposition, the solid is filtered and washed it with a small amount of acetic anhydride and simple ether. After drying in a vacuum dryer receive 4,28 g (58.2 per cent) of the target product.

The identity of the obtained product is confirmed by the range of1H and13Nuclear magnetic resonance and infrared spectrum and its purity determined by high-performance liquid chromatography.

Elemental analysis of the product for C14H26Cl2N2O4Pt:

Found, %: C 30,24; H 4,75; N 4,99; Cl 12,81.

Calculated, %: C 30,44; H 4,74; N 5,07; Cl 12,84.

Example 2

Synthesis of complex AF-bis (acetato)-b-(1-amino-3,5-dimethyl-adamantane)-Amin de dichloroplatinum (IV)) (hereinafter called "LA-15") [(OS-6-43)-bis(acetato)(1-amino-3,5-dimethyl-adamantane) Emmendingen (IV)]

Mix 0.96 g (1/93 mmol) of b-(1-amino-3,5-dimethyladamantane) with a room temperature with an excess of acetic anhydride (8 ml, 84,7 mmol). After dissolution, the reaction mixture is added 10 ml of a simple ester. Stirring is continued until the ends of the sediment. The precipitated solid is filtered, washed with simple ether and dried in a vacuum dryer. The output is 0,72 g (64,3%) of the target product.

The identity of the obtained product is confirmed by the range of1H and13Nuclear magnetic resonance and infrared spectrum and its purity determined by high-performance liquid chromatography.

Elemental analysis of the product for C16H30Cl2N2O4Pt:

Found, %: C 32,88; H To 5.21; N 4,75; Cl 12,31.

Calculated, %: C 33,11; H To 5.21; N A 4.83; Cl 12,22.

Example 2A

Synthesis of b-(1-adamantylamine)-Amin-de-dichloro-AF-dihydroxylation (IV)) (hereinafter called LA-11)

Suspended 8,01 g (18,44 mol) complex CIS-(1-adamantylamine)-Amin-dichloroplatinum (II) in 120 ml of water at room temperature. To the suspension is added a stoichiometric excess (20 ml), 30% (weight/weight) aqueous solution of hydrogen peroxide, the reaction mixture is heated at 80oC for 1 hour and then cooled to room temperature. The solid is separated by filtration, prominade of dimethylformamide is removed by simple washing with ether. After drying in a vacuum dryer output is 6,45 g, i.e. 74.6% of theoretical (with respect to the original complex of platinum (II)).

The identity of the product confirm infrared spectral analysis and the purity determined by high-performance liquid chromatography.

Elemental analysis of the obtained product (for C10H22Cl2N2O2Pt):

Found, %: C 25,75; H Amounts To 4.76; N 5,94; Cl 15,10.

Calculated, %: C 25,65; H 4,74; N 5,98; Cl 15,14.

Example 3

Synthesis of complex inclusion compounds LA-12 with hydroxypropyl-beta-cyclodextrin (hereinafter called "dosage form include LA-12")

Connection LA-12 was dissolved in acetone to obtain a final concentration of 20 g/l To the resulting solution was added hydroxypropyl-beta-cyclodextrin (164 g/l). To this mixture is added slowly buffered aqueous phase of 100 mm Hepes with pH of 7.3 with stirring at room temperature until the final volume ratio of acetone and the aqueous phase becomes equal to 1:10. Dissolved cyclodextrin dissolves quickly even after the first additions of the aqueous phase (up to 10% of the total volume of the aqueous phase). Finally, acetone and water are removed from the solution of the complex include liofilizirovannam of these compounds in the in vivo studies in animals with experimental tumors, such as murine leukemia L1210 in ascitic form (IP L1210) or in solid form (SC-L1210) grown on DBA2 mice, Gardner lymphosarcoma in a solid and ascitic form (LsG), grown in mice SN, tumor Ehrlich (Ehrlich) in solid (STE) and ascitic form (ATE) grown on NMRI mice, the tumor MS (transplantable adenocarcinoma mammals), grown in DBA1 mice, the metastatic melanocarcinoma B16 in solid and ascitic form (SC-B16, IP-B16), grown in mice SW, and metastatic carcinoma, Lewis lung (LL), grown in mice SV, namely according to the method of V. Jelinek (Neoplasma 12, 469 (1965), ibid 7,146 (1960)).

Estimated time of survival and expected optimal dose, in addition to point definitions, the estimated t-Student test, proportional risk model Coke (SOH) and the method of Carter (Carter W. H. et al.: Cancer Res. 42, 2963 (1982)). Curves activity obtained by this method of calculation allows to calculate the optimal dose and to establish the toxic dose. This method allows to evaluate not only the effect with monotherapy, but also the General toxicity and the influence of the components of the combination therapy. Therefore, the relationship between dose and activity were modeled and evaluated in some instances also by this method.

For C is of biological interest, having a tumor, namely the inhibition of tumor growth and inhibition of reduction of time of survival of a biological object. For the tumor growth can be observed both clinically and in experiments in vivo, i.e. in experimental animals. Evaluation of tumor growth can be made by weighing the tumor tissue or by measurement of tumor size.

Such a positive therapeutic effect of the compounds of the present invention on mice female strain SN has been proven to reduce tumor mass in animals with a solid form of lymphosarcoma Gardner (SC-LsG), with STE, SC-B16, SC-LL and S, it is by oral administration of the compounds of this group of chemicals. It has been shown statistically significant (P0,05) or even highly reliable (P0,01) reduction in the average weight of the tumors compared with the control, i.e., the untreated group.

Also, the compounds of the present invention prolong the survival time of the relevant biological objects, for example, mice with tumors IP L1210, MS, SC-L1210, IP-LsG and ATE, and also extremely with IP-B16 and IP LsG, it is by oral administration. Because, by their nature visionportfolio test systems lead to a fatal outcome, the antitumor effect of the compounds on izopet is treated control animals. In typical experiments (see examples) one experimental group usually consisted of 10 animals, and the animals of the treated groups survived longer statistically compared with the control, i.e., untreated animals.

To use their antitumor activity of the disclosed compounds can enter the relevant biological objects, namely mammals, in usable form, and the usual routes of administration. They can be administered alone or, preferably, as the active substances together with any suitable non-toxic pharmaceutical carrier, dissolved or suspended, for example, water, buffer, saline solution, in the solution of methylcellulose, polyethylene glycol, polypropyleneglycol etc. Oral administration is the most preferred. The optimal dose depends on the type of tumor that is being treated, the type of biological object that is treated, from its weight and/or body surface, the location of the tumor, on its morphological type from the schema doses, etc. From the biological experiments it is obvious that, for example, an effective single oral dose of injected LA-12 is only 10 mg/kg (30 mg/m2for IP doses of LA-12, the optimal dose for DBA2 mice with leukemia L1210 was 22.9 mg/kg orally 3 (of 68.7 mg/m23) and with the continuous introduction (1-9 day) the optimal dose for the same mice is 9.6 mg/kg oral 9 (28,8 mg/m29).

The toxicity of the compounds according to the invention is low: the value LD50for NMRI mice may be above 600 mg/kg orally.

It is obvious that we can expect useful therapeutic effects at doses that are absolutely non-toxic to mammals. According to research conducted biological tests can be expected that portable and effective single oral dose for humans would be at the level of 30 mg/m2.

In subsequent tests, which will show the antitumor activity of compounds of the present invention, as a standard, we used the following compounds of the prior art: the complex AF-bis(acetato)-b-Ammin-CD-dichloro-s- (cyclohexylamine) platinum (IV) [(OS-6-43)-bis(acetato)] ammendale (cyclohexylamine) platinum (IV)] (JM216, Johnson Matthey Technology Centre, Reafing, Berkshire, Great Britain, Kelland et al., 1993), referred to as compound LA-2 and the complex CIS-diamminedichloroplatinum (II), which is the active substance Medi complex AF-bis (acetato)-b-(1-adamantylamine)-Amin de dichloroplatinum (IV) (compound (I), A=1-adamantylamine, "LA-12") in mice after a single oral administration and comparison with complex AF-bis (acetato)-b-Ammin-CD-dichloro-s-(cyclohexylamine) platinum (IV) (compound (I), And=cyclohexylamine, code JM216-Johnson Matthey Technology Centre, "LA-2") and with the complex CIS-diamminedichloroplatinum (II) (NSC 119875, the active drug substance of Platydema 10 injecting dry)

150 mice female DBA2 strain, with an estimated mass of 18 g were divided into fourteen groups: one control group (21 animal) and 13 experimental groups (9-10 animals). All the animals intraperitoneally was inoculable lethal dose of ascitic fluid from leukemia L-1210. Animals 10 experimental groups were treated with compounds LA-12 and LA-2 in aqueous suspensions prepared ad hoc immediately before use. The suspension had the following number of relevant connections that experimental animals received one day after inoculation of the tumor dose of 160, 80, 40, 20 and 10 mg/kg, respectively, in amounts of 0.2-0.4 ml of Platinium animals were injected three groups subcutaneously in the form of an isotonic aqueous solution, prepared by dissolving liofilizirovannogo powder in water for injection immediately before use. The animals were monitored to determine survival time. Dependence snowline time points in response to biological reactions was evaluated test the equivalence of the two middle values (t-test Student) under the assumption of logarithmic/normal nature of the distribution of time values and the assumption about the possible unknown variations (Roth et al., 1962). The geometric mean value was calculated from the individual values of survival time. The average values of the criterion test, the difference exceeding a critical value at a significance level of 5% was evaluated as statistically significant.

Observed that animals treated with LA-12, there was a statistically significant (t-test, P0,05) higher average survival time in the group with a dose of 10 mg/kg oral, namely 55% compared with the untreated control group. Platinum, at the dose of 5 mg/kg subcutaneously, increased the average survival time by 32% in comparison with controls, but this difference in values was not statistically significant in comparison with the control group. The effect has not been proven to LA-2.

The results are summarized in table. 1.

Mouse DBA2, females, 16,6-19,1, the Transplantation of the tumor intraperitoneal introduction, inoculate 106tumor cells. Start of treatment 1 day after transplantation (1 oral, Platinium 1 subcutaneously 1 day).

In table. 1 lists relative to the corresponding doses of the average values of survival time, the confidence intervals of the geometric mean for P=1-=0.95 and is suitable is>/BR>Illustration of antitumor activity of complex AF-bis(acetato)-b-(1-adamantylamine)-Amin de dichloroplatinum (IV) (compound (I), A=1-adamantylamine, "LA-12") in mice after oral fractional introduction and comparison with the complex AF-bis(acetato)-b-Ammin-CD-dichloro-s-(cyclohexylamine)platinum (IV) (compound (I), And=cyclohexylamine, code JM216 "LA-2"-Kelland et al., 1993) and with the complex CIS-diamminedichloroplatinum (II) (NSC 119875, the active drug substance of Platydema 10 injecting dry)

In a similar experiment on animals with leukemia L1210 compound (I) (A=adamantylamine, LA-12) was administered repeatedly in three daily doses, namely, 1-, 4 - and 9-th day after inoculation, together with LA-2 oral and Platydema subcutaneously according to the same scheme.

Observed that animals treated with LA-12 was statistically significant (t-test, P0,05) higher average survival time in the group with a dose of 4 mg/kg/day orally 3 (148%) in comparison with the untreated control group. Connection LA-2 showed the same effect (148%) only in double dose, i.e. the dose of 8 mg/kg/day oral 3. The optimal dose of LA-12, designed by Carter (Carter), amounted to only 4,55 mg/kg/day orally 3, while for LA-2, it was equal 10,96 mg/kg/day orally effektivnye, than a typical connection of the above-mentioned patents on its optimal therapeutic scheme.

The results are summarized in table. 2.

Mouse DBA2, females, 19,8-21,2, the Transplantation of the tumor intraperitoneal introduction, inoculate 106tumor cells. Start of treatment 1 day after transplantation (3 oral, Platinum subcutaneously 3 1-, 5 - and 9-th day).

In table. table 2 lists, in relation to the relevant doses, the average time of survival, the confidence intervals of the geometric mean for P= 1-=0.95 and the corresponding values of the average time of survival in the percentage of those in the control group.

Example 6

Illustration of antitumor activity of complex AF-bis(acetato)-b-(1-adamantylamine)-Amin de dichloroplatinum (IV) (compound (I), A=1-adamantylamine, "LA-12") in mice with leukemia L1210 after continuous and fractional oral administration and comparison with complex AF-bis(acetato)-b-Ammin-cd-dichloro-s-(cyclohexylamine) platinum (IV) (compound (I), And=cyclohexylamine, code JM216 "LA-2"-Kelland et al., 1993) and with the complex CIS-diamminedichloroplatinum (II) (NSC 119875, the active drug substance of Platydema 10 injecting dry)

In the experiment on animals with leukemia L1210, CH cases in 9 daily doses namely, 1-9 th day after inoculation, and also fractional 1-, 4 - and 9-day together with LA-2 oral so that the total dose were similar for both schemes. Platinum was administered subcutaneously.

Connection LA-12 statistically significantly increased the value of the survival time compared with the control group at a dose of 6 mg/kg oral 9, namely, 130%, and it has been assessed as having antitumor activity similar to Placidia at doses of 8 and 4 mg/kg subcutaneously 1. Connection LA-2 was not increased statistically significantly the survival time compared with the control group in any of the groups, doses, and, therefore, it was assessed as not having antitumor activity.

When fractional scheme (1-, 5 - and 9-th day after inoculation) effect has not been proven by point definitions.

Assessment according to the dose/effect of LA-12 showed that continuous treatment is more predominant when the same total dose.

The results are summarized in table. 3.

Mouse DBA2, females, 19,3-21,4, the Inoculation of the tumor intraperitoneal introduction, inoculate 106tumor cells. Start of treatment 1 day after inoculation (9 oral, 1-9-nd and 3 1-, 5 - and 9-day catfish doses, mean values of survival time, the confidence intervals of the geometric mean for P= 1-=0.95 and the corresponding values of the average time of survival in the percentage of those in the control group.

Example 7

Illustration of antitumor activity of complex AF-bis(acetato)-b-(1-adamantylamine)-Amin de dichloroplatinum (IV) (compound (I), A=1-adamantylamine, "LA-12"), dosage forms include LA-12 and complex AF-bis(acetato)-b-(1-amino-3,5-dimethyladamantane)-Amin de dichloroplatinum (IV) (compound (I), A=1-amino-3,5-dimethyladamantane, "LA-15") in mice with leukemia L1210 after continuous oral administration and comparison with complex AF-bis(acetato)-b-Ammin-cd-dichloro-s- (cyclohexylamine)platinum (IV) (compound (I), And=cyclohexylamine, code JM216 "LA-2"-Kelland et al., 1993) and with the complex CIS-diamminedichloroplatinum (II) (NSC 119875, the active drug substance of Platydema 10 injecting dry)

In the experiment on animals with leukemia L1210 carried out analogously to examples 4, 5 and 6, the connection LA-12, new dosage form, the compound LA-15 was introduced repeatedly in nine daily doses, namely 1-9-th day after inoculation, together with LA-2 orally, according to the same scheme. Platinum was administered subcutaneously.

Connect the STU compared with the control group used in the dose namely, 130%, and they were assessed as having antitumor activity similar to Placidia at a dose of 4 mg/kg subcutaneously 1 (123%) and LA-2 at a dose of 3 mg/kg/day orally 9 (114%).

The results are summarized in table. 4.

Mouse DBA2, females, 18,0-19,7, the Inoculation of the tumor intraperitoneal introduction, inoculate 1,2106tumor cells. Start of treatment 1 day after inoculation (9 oral 1-9 day, Platinium 1 subcutaneously 1 day).

In table. 4 lists, in relation to the relevant doses, the average time of survival, the confidence intervals of the geometric mean for P= 1-=0.95 and the corresponding values of the average time of survival in the percentage of those in the control group.

Example 8

Illustration of antitumor activity of complex AF-bis(acetato)-b-(1-adamantylamine)-Amin de dichloroplatinum (IV) (compound (I), A=1-adamantylamine, "LA-12"] in mice with Gardner lymphosarcoma after a single oral administration and comparison with complex AF-bis(acetato)-b-Ammin-cd-dichloro-s-(cyclohexylamine) platinum (IV) (compound (I), And=cyclohexylamine, code JM216 "LA-2"-Kelland et al., 1993) and with the complex CIS-diamminedichloroplatinum (II) (NSC 119875, the active drug substance Platinumgroup, namely one control and nine experimental groups, each containing 20 animals. All animals transplanted lethal dose of tumor homogenate from Gardner lymphosarcoma. Animals of the experimental groups were treated with compounds LA-12 and LA-2 in aqueous suspensions prepared ad hoc immediately before use. The suspension contained such amounts of the respective compounds, which experimental animals were received on the 5th day after inoculation of the tumor dose is 32.8 and 2 mg/kg, respectively, in amounts of 0.2-0.4 ml; Platinum used a single dose of 8.4 and 2 mg/kg subcutaneously. Half of the animals in the group with each dose were killed on the 14th day after inoculation under ether anesthesia and tumors were surgically removed from the body. When the weighting was determined by the tumor weight of each animal. The second half of the animals were left for monitoring survival time.

Observed that animals treated with LA-12 was statistically significant (t-test, P0,05) lower average tumor weight (77%) at a dose of 32 mg/kg orally in comparison with the control group (100%). The effect was not observed with LA-2, the connection of the prior art.

The results are summarized in table.5 .

Mouse SN, females, 20,7-28,3, Transplantation, Platinium 1 subcutaneously 5th day).

In table. 5 lists, in relation to the relevant doses, the average weight of tumors, confidence intervals arithmetic mean of the values for P=1-=0.95 and the corresponding values of the average tumor mass in the form of a percentage from that in the control group.

Example 9

Illustration of antitumor activity of complex AF-bis(acetato)-b-(1-adamantylamine)-Amin de dichloroplatinum (IV) (compound (I), A=1-adamantylamine, "LA-12") in mice with adenocarcinoma MS mammals after repeated continuous oral administration and comparison with complex AF-bis(acetato)-b-Ammin-CD-dichloro-s-(cyclohexylamine)platinum (IV) (compound (I), And=cyclohexylamine, code JM216 "LA-2"-Kelland et al., 1993) and with the complex CIS-diamminedichloroplatinum (II) (NSC 119875, the active drug substance of Platydema 10 injecting dry)

130 mice females of strain DBA1 weighing approximately 20 g were divided into 12 groups, namely one control and eleven experimental groups, each containing 10 animals. All animals were subcutaneously inoculable lethal dose of tumor homogenate. Animals of the experimental groups were treated with compounds LA-2, LA-12 and LA-15 in aqueous suspensions prepared ad hoc directly who's received a dose of 12.6 and 3 mg/kg/day orally 9 respectively in amounts of 0.2-0.4 ml; Platinum used a single dose of 8 and 4 mg/kg subcutaneously in a volume of 0.2 and 0.4 ml on the fifth day after inoculation.

Animals from the control group and from each dose group were weighed and determined the sizes of the tumors on the 14th day after inoculation. Half of the animals from the control group were killed under ether anesthesia and tumor surgically removed from the body. The dependence of the tumor mass from its size was estimated by regression analysis. The tumor weight of each animal was determined by this regression function. The weight of tumors was evaluated in the case of point definitions test the equivalence of the two middle values (t-test Student) under the assumption of normal distribution of mass values and the assumption about the possible unknown variations. The arithmetic average value was determined for the individual values of the tumor mass. The differences in average values at which the value of the test criterion was exceeded the critical value at a significance level of 5% was considered statistically significant.

The dependence of the survival time from the used dose was evaluated in comparison with untreated control. Time was evaluated similarly as in the cases of leukemia and of ascitic tumor is under the assumption of logarithmic/normal nature of the distribution of time values and the assumption about the possible unknown variations (Roth Z., Josifko M. , V. Maly, Trcka V.: Statisticke metody v experimentalni medicine, SZN Prague, 1992, p. 278). The geometric mean was calculated for the individual values of time of survival. The average values of the criterion test, differences which exceeded the critical value at a significance level of 5% was evaluated as statistically significant.

Observed that animals treated with LA-12 was statistically significant (t-test, P0,05) lower average weight of tumors (86%) at the dose of 3 mg/kg/day orally 9. In addition, they were statistically significant (t-test, P0,05) higher mean values of survival time (128%) in comparison with the untreated control group (100%). Other tested compounds (except Platydema) were not statistically significant differences in point definitions survival time in comparison with the control group.

Connection LA-12 and Platinum, was assessed as having antitumor activity. The effect was not observed with LA-2, the connection of the prior art.

The results are summarized in table. 6-1.

Mouse DBA1, females, 20,4-21,8, the Inoculation of the tumor subcutaneous injection of 0.2 ml of homogenate tumors, dilution 1:1. Start of treatment with 5-th day after InoculateIT doses, mean values of survival time, the confidence intervals of the geometric mean for P=1-=0.95 and the relative values of the average time of survival in the percentage of those in the control group.

Mouse DBA1, females, 20,4-21,8, the Inoculation of the tumor subcutaneous injection of 0.2 ml of homogenate tumors, dilution 1:1. Start of treatment with 5-th day after inoculation (9 oral, 5-13 day, Platinium 1 subcutaneously 5th day).

In table. 6-2 lists, in relation to the relevant doses, the average weight of tumors, confidence intervals arithmetic mean of the values for P=1-=0.95 and the corresponding values of the average tumor mass in the form of a percentage from that in the control group.

Example 10

Illustration of antitumor activity of complex AF-bis(acetato)-b-(1-adamantylamine)-Amin de dichloroplatinum (IV) (compound (I), A=1-adamantylamine, "LA-12") in mice with ascitic tumor Ehrlich (Ehrlich) after repeated oral administration and comparison with complex AF-bis(acetato)-b-Ammin-CD-dichloro-s-(cyclohexylamine)platinum (IV) (compound (I), And=cyclohexylamine, code JM216 "LA-2"-Kelland et al., 1993) and with the complex CIS-diamminedichloroplatinum (II) (NSC 119875, the active drug substance of Platydema 10 Indo control and nine experienced, each containing 20 animals. All the animals intraperitoneally was inoculable lethal dose of ascitic fluid from the ATE. Animals of the experimental groups were treated with compounds LA-12 and LA-2 in aqueous suspensions prepared ad hoc immediately before use. The suspension had the following number of relevant connections that experimental animals received doses of 12, 6 and 3 mg/kg/day orally 9 respectively in amounts of 0.2-0.4 ml; Platinum used a single dose of 8, 4 and 2 mg/kg subcutaneously in a volume of 0.2 and 0.4 ml on the first day after inoculation. Half of the animals in each dose group were killed on the 10th day after inoculation under ether anesthesia, after laparotomy was aspirated ascitic fluid and determined the mass of tumors in each animal by difference in weight before and after extraction of the ascitic fluid. The remaining animals were left for monitoring survival time.

Observed that the treated animals had lower average tumor weight compared with the untreated control (table. 7-1). However, the increase in the average survival time was statistically highly significant (P0,01) compared with the untreated control only for LA-12 and Platydema, but not LA-2 (PL. 7-2).

Mouse ICR, female, with 23.7-25, after inoculation (9 oral, 1-9 day, Platinium 1 subcutaneously 1 day).

In table. 7-1 lists, in relation to the relevant doses, the average weight of tumors, confidence intervals arithmetic mean of the values for P=1-=0.95 and the corresponding values of the average tumor mass in the form of a percentage from that in the control group.

Mouse ICR, female, with 23.7-25,4, the Inoculation of the tumor intraperitoneal introduction, inoculate 0.2 ml of ascitic fluid. Start of treatment 1 day after inoculation (9 oral, 1-9 th days, Platinium 1 subcutaneously 1 day).

In table. 7-2 lists, in relation to the relevant doses, the average time of survival, the confidence intervals of the geometric mean for P= 1-=0.95 and the corresponding values of the average time of survival in the percentage of those in the control group.

Illustration of antitumor activity of complex AF-bis(acetato)-b-(1-adamantylamine)-Amin de dichloroplatinum (IV) (compound (I), A=1-adamantylamine, "LA-12"), dosage forms include LA-12 and complex AF-bis(acetato)-b-(1-amino-3,5-dimethyladamantane)-Amin de dichloroplatinum (IV) (compound (I), A=1-amino-3,5-dimethyladamantane, "LA-15") in mice ATE after continuous and fractional oral is And=cyclohexylamine, code JM216, "LA-2"-Kelland et al., 1993) and with the complex CIS-diamminedichloroplatinum (II) (NSC 119875, the active drug substance of Platydema 10 injecting dry)

In the experiment on the animals ATE, which was set analogously to example 10, compound LA-12, its dosage form of inclusion and connection LA-15 was introduced repeatedly in nine daily doses, namely 1-9-th day after inoculation with LA-2 oral according to the same scheme. Platinum was administered subcutaneously.

After the application, half of the animals in each dose group were killed on the 10th day after inoculation under ether anesthesia, after laparotomy ascitic fluid was aspirated and has determined the mass of the tumor in each animal according to the difference of mass before and after the extraction of the ascitic fluid. Determined antitoxic in ascitic fluid of individual animals to the centrifuge micromethod determine hematocrit capillary tubes with heparin length of 75 mm Is "common antitoksica", corresponding to the mass of cells in the ascitic fluid was determined by the percentage of antitoksica and tumor mass. The remaining animals were kept for monitoring survival time. In these animals, the survival time was controlled every day. The dependence of the values of survival time from opalocka, as a biological response, evaluated test the equivalence of the two middle values (t-Student test). The geometric mean value was determined for the individual values of time of survival, the arithmetic mean value was calculated from the individual values of the tumor mass and total antitoksica. The difference between the mean values was assessed by a test of the equivalence of the two middle values (t-test Student) under the assumption of a normal probability distribution of mass values and values shared antitoksica and logarithmic/normal distribution of time values, all at the suggestion of a possible unknown variations. The differences between the average values, whose values criterion test exceeded the critical value at a significance level of 5 and 1%, were evaluated as statistically significant and highly statistically significant, respectively.

The optimal dose was calculated from the values of time of survival under proportional risk model Coke (SOH) (Carter et al., 1982). The distribution parameters Weinbull and logarithmic transformation of the preliminary data used to establish the basic and modified functions of survival from experimental data.

Observed that the treated animals had smaller average tumor mass compared to untreated control group (table. 8-2), but this difference was not statistically significant.

The average mass of the cellular fraction of the tumor (total ascitic) was statistically significantly lower only in the case of compound LA-15 at a dose of 6 mg/kg/day orally 9 (PL. 8-3), what is the evidence of antitumor activity of the compounds.

The results obtained are summarized in table. 8-1 and 8-2.

Mouse ICR, female, 30,1-33,7, the Inoculation of the tumor intraperitoneal introduction, inoculate 5102tumor cells in 0.2 ml of ascitic fluid. Start of treatment 1 day after inoculation (9 oral, 1-9 day, Platinium 1 subcutaneously 1 day).

In table. 8-1 lists, in relation to the relevant doses, the average time of survival, the confidence intervals of the geometric mean for P= 1-=0.95 and the corresponding values of the average time of survival in the form of proceedinas, the inoculate 5106tumor cells in 0.2 ml of ascitic fluid. Start of treatment 1 day after inoculation (9 oral, 1-9 day, Platinium 1 subcutaneously 1 day).

In table. 8-2 lists, in relation to the relevant doses, the average weight of tumors, confidence intervals arithmetic mean of the values for P=1-'=0.95 and the corresponding values of the average weight of tumors in the form of a percentage from that in the control group.

Mouse ICR, female, 30,1-33,7, the Inoculation of the tumor intraperitoneal introduction, inoculate 5106tumor cells in 0.2 ml of ascitic fluid. Start of treatment 1 day after inoculation (9 oral, 1-9 day, Platinium 1 subcutaneously 1 day).

In table. 8-3 lists, in relation to the relevant doses, the average weight of tumors, confidence intervals arithmetic mean of the values for P=1-=0.95 and the corresponding values of the average weight of tumors in the form of a percentage from that in the control group.

1. The complex compound of platinum with oxidation state IV formula I

< / BR>
where X represents a halogen atom;

In represent independently from each other hydroxyl group or a carboxylate group containing 1-6 atoms angle is Asim 10-14 carbon atoms, which may be optionally substituted in the tricyclic ring by one or two alkyl group(s), each containing 1-4 carbon atoms.

2. The complex compound of platinum under item 1 of the formula I, where a represents adamantylamine and y have the meanings given in paragraph 1.

3. The complex compound of platinum under item 1 of the formula I, where a is 3,5-dimethyladamantane and y have the meanings given in paragraph 1.

4. Complex inclusion complex of platinum with oxidation state IV formula I

< / BR>
where X represents a halogen atom;

In represent independently from each other hydroxyl group or a carboxylate group containing 1-6 carbon atoms;

A represents a group-NH2-R, where R is a tricyclic hydrocarbon fragment containing 10 to 14 carbon atoms, which may be optionally substituted in the tricyclic ring by one or two alkyl group(s), each containing 1-4 carbon atoms,

with beta - or gamma-cyclodextrin, which may be optionally substituted hydroxyalkyl groups containing 1-6 carbon atoms.

5. The method of obtaining complex platinum compounds of the formula I on p. 1, Otie in p. 1,

oxidized on the platinum atom hydrogen peroxide in the formation of dihydrochalcones platinum (IV) and, optionally, a hydroxyl group specified complex replaces the carboxylate groups by the action Alliluyeva agent.

6. The way to obtain the inclusion complex of platinum complex of formula I, beta - or gamma-cyclodextrin, which may be optionally substituted hydroxyalkyl groups containing 1-6 carbon atoms, under item 4, characterized in that the solution of the platinum complex of formula I in an organic solvent is mixed with an aqueous solution of beta - or gamma-cyclodextrin, which is optionally substituted hydroxyalkyl groups containing 1-6 carbon atoms, with subsequent evaporation of the solvent from the resulting solution.

7. Complex platinum compound of formula I or its inclusion complex with beta - or gamma-cyclodextrin under item 1 or 4 as a pharmaceutical product with antitumor activity.

8. Pharmaceutical composition for treatment of cancer, characterized in that it contains at least one complex of the platinum compound of formula I or its inclusion complex with beta - or ambient.

 

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