MXPA98000129A - 2-aminocarbonil-1, 2-bis (metilsulfonil) -1- (substitute) hydrains antitumora - Google Patents

Abstract

The present invention relates to novel 2-aminocarbonyl-1,2-bis (methylsulfonyl) -1- (2-chloroethyl) hydrazines and 2-aminocarbonyl-1,2-bis (methylsulfonyl) -1-methylhydrazines, and their use for treat malignant tumors The agents are especially useful in the treatment of cancers of animals and humans. The two preferred agents in this class, especially for use in the treatment of tumors are 2-aminocarbonyl-1,2-bis (methylsulfonyl) -1- (2-chloroethyl) -2- (2-chloroethyl) aminocarbonylhydrazine and 1, 2-bis (methylsulfonyl) -1- (2-chloroethyl) -2-methylaminocarbonylhydrazine. These agents are characterized by the following: they are incapable of inactivation by the proposed denitrosation mechanism for the inactivation of the CNU, they are incapable of generating a hydroxyethylation species by the mechanism proposed for the CNU, and they are capable of chloroethylation or methylation and carbamoylation

Description

2-AMIN0CARBTNIL-1, 2-BI S (METÍLSULFO IL) - 1 - (SUBSTITUTE) ANTI-TUMOR HYDRAINS.

FIELD OF THE INVENTION The present invention relates to 2-aminocarbonyl-1,2-bis (methylsulfonyl) -1- (substituted) hydrazines which exhibit antitumor activity in mammals. Methods for treating neoplasia, especially including solid tumors, are additional aspects of the present invention. This work was supported by a grant from the National Institutes of Health. The Government of the United States of North America maintains certain rights in the invention.

BACKGROUND OF THE INVENTION The search for improved anti-neoplastic activity display compounds is focused on some attention to nitrosourea compounds such as 1,3-bis (2-chloroethyl) -1-nitrosourea (BCNU) and related agents. Several N- (2-chloroethyl) -N-nitrosoureas (CNUs) have been evaluated in a clinical manner and shown to have activity REF: 26672 significant antineoplastic against brain tumors, colon cancer and liposomes (see, DeVita, et al, Cancer Res. 1965, 25, 1876-1881; Nissen, et al., Cancer 1979, 43, 31-40). The characterization of the decomposition products of the clinically used BCNUs, such as BCNU and 1- (2-chloroethyl) -3-cycloexyl-1-nitrosourea (CCNU), has resulted in the identification of several reactive products, including species of chloroethylation, carbamoylation and hydroxytylation (see, for example, Montgomery, er al., J. Med. Chem. 1967, 10, 668-674; Montgomery, et al., J. Med. Chem. 1975, 18, 568-571. ein am and Lin, J. Med. Chem. 1979, 22, 1193-1198, and Brundrett, RB, J. Med. Chem. 1980, 23, 1245-1247). The antitumor activity of CNUs has been suggested to result from chloroethylation and subsequent DNA cross-linking (see, Kohn, KW in Recent Results in Cancer Research (Eds. Carter, SK, Sakarai, Y., and Umeza a, H. ), vol 76, p.141, Springer, Berlin (1981)). A support of this observation is that many chloroethylating agents without carbamoylation activity (see, clomesone, as discussed by Shealy, et al., J. Med. Chem. 1984, 27, 664-670) possess excellent antineoplastic activity. In addition, replacement of the chloro group in the CNU for a hydroxyl group has resulted in a considerable decrease in antineoplastic activity (Montgomery, JA, personal communication, cited by Gibson, et al., Cancer Res. 1986, 46, 553-557). Additionally, there is some evidence that hydroxyethylation of DNA is a carcinogenic and / or mutagenic case (Pelfrene, et al., J. Nati, Cancer Inst. 1976, 56, 445-446; and Swenson, et al., J. Nati. , Cancer Inst. 1979, 63, 1469-1473). While hydroxyethylation does not seem to have a salutary effect on the antineoplastic activity of the CNU, there seems to be some uncertainty regarding the role played by the carbamoylation species (ie isocyanate). The isocyanate generated from the CNU reacts with the thiol and amine functionalities in the proteins and inhibits the DNA polymerase (Baril, et al., Cancer Res. 1975, 35, 1-5.), Repair of the strand of DNA breaks (Kann, et al., Cancer Res. 1974m 34m 398-402), and the synthesis and processing of RNA (Kann, et al., Cancer Res. 1974, 34, 1982-1988).

In addition, BCNU has been shown to be glutathione reductase, ribonucleotide reductase and thioredoxin reductase (Schallreuter, et al., Biochim Biophys. Acta 1990, 1054, 14-20). Although it is believed by many that some of these properties contribute to the toxic side effects of CNU (Colvin, et al., Biochem Pharmacol, 1976, 25, 695-699; heeler, et al., Cancer Res. , 34, 194-200; and Panasci, et al., Cancer Res. 1977, 37, 2615-2618), is entirely possible, as speculated by Gibson and Hickman (Gibson and Hickman, Biochem. Pharmacol., 1982, 31 , 2795-2800) in their study of the effects of BCNU on the TLX tumor in mice, that the intracellular release of isocyanates plays a role in modulating the biological activity of CNU against some specific tumor types. Caracemide, a research antitumor agent, developed by Dow Chemical Company (New an and Farquhar, Invest, New Drugs 1987, 5, 267-271 and Slatter, et al., Chem. Re. Toxicol., 1993, 6, 335-340) it is thought to act as a latent form of methyl isocyanate. This agent was shown to be active in a number of tumor models from the National Cancer Institute, including the human breast cancer MX-1 and CX-1 human cenoingertos, implanted in subreal capsules of athymic mice (clinical journal "Carace ide"). NSC 253272", Division of Cancer Treatment, National Cancer Institute, 1983).

The hydroxyethylation species generated from the CNU, 2-hydroxyethyldiazohydroxide, is thought to be formed of 4,5-dihydro-1,2,3-oxadiol, which, in turn, has been hypothesized to be the result of an internal cyclization reaction comprising the N-nitroso group (Brundrett, RB, J. Med. Chem. 1980, 23, 1245-1247). The N-nitroso group is also included in the enzymatic inactivation of the CNU. For example, BCNU can be inactivated by denitrozation by the nitrosomic enzymes of the liver in the NADPH-dependent reaction, with the formation of 1,3-bis (2-chloroethyl) urea (Hill, et al., Cancer Res. 1975 , 35, 296-301 and Lin and Weinkam, J. Med. Chem. 1981, 24, 761-763). The denitrozation reaction is catalyzed by NADPH P450 reductase: cytochrome in the case of CCNU (Potter and Reed, Arch. Biochem. Biophys., 1982, 216, 158-169 and Potter and Reed, J. Biol. Chem. 1983, 258, 6906-6911). The BCNU has also been shown to undergo glutathione-dependent denitrozation catalyzed by the glutathione S-transferan BCNU isoenzymes of the rat (Smith, et al., Cancer Res. 1989, 49, 2621-2625) and human (Berhane, et al. ., Cancer Res. 1993, 53, 4257- Since denitrozation catalyzed by tumor cells could be a potential mechanism of resistance to the CNU, the present purpose was to synthesize a series of compounds that (a) were capable of generating a species of chloroethylation or methylation, (b) they were capable of forming a kind of carbamoylation, and (c) they were devoid of hydroxyethylation activity, and (d) they were free of the structural characteristics that would make them highly prone to metabolic inactivation. It is believed that the 2-aminocarbonyl-l, 2-bis (methylsulfonyl) -l- (substituted) hydrazines (I) could satisfy the above conditions for the following reasons: (a) catalyzed elimination based on the compounds I would result in the formation of a chloroethylation or methylation species and a carbamoylating agent as shown below. * CH3ßOa-H-M80aCH3 - > OT3B02IMp- + RHCO + CH3SOaH C tf-B II O H I (Y is methyl or 2-chloroethyl) (b) at least three classes of prodrugs of the species, ie 1,2-bis (methylsulfonyl) -1- (2-chloroethyl) hirazine, 1- (2-cycloethyl) -1,2,2-tris (methylsulfonyl) hydrazine (Shyam, et al., J. Med. Chem. 1990, 33, 2259-2264), and l-acyl-1,2-bis (methylsulfonyl) -2- (2-chloroethyl) hydrazine (Shyam, et al., J. Med. Chem. 1993, 36, 3496-3502), with potent antitumor activity, have been identified. (c) the formation of an intermediate of 4,5-hydroxy-1,2,3-oxadiazole can be prevented by the absence of a portion of N-nitroso. In this way, in turn, the formation of a 2-hydroxyethylation agent can be prevented. The absence of an N-nitroso group can also make the compounds less prone to metabolic inactivation.

OBJECTS OF THE INVENTION It is an object of the invention to provide effective antineoplastic agents for the treatment of numerous cancerous conditions, including solid tumors in animals and humans. It is another object of the invention to provide antineoplastic agents that are capable of generating a chemical species of carbamoylation and chloroethylation. It is still a further object of the invention to provide effective antineoplastic agents that have been less prone to metabolic inactivation than the compounds of the related structure. It is a further object of the invention to provide pharmaceutical compositions based on the use of the new antineoplastic agents. It is still another object of the invention to provide methods for treating neoplasia, including solid tumors, in animals and humans. These and other objects of the invention can easily be assembled from the description of the invention as follows.

BRIEF DESCRIPTION OF THE INVENTION The present invention relates to 2-aminocarbonyl-l, 2-bis (methylsulfonyl) -1- (substituted) hydrazines compounds of the formula: CH3S02N (Y) N (CONHR) S02CH3 where Y is -CH3 or -CH2CH2C1; and R is alkyl of 1 to 7 carbon atoms, cyclohexyl, methylcyclohexyl, CH2CH = CH2, -CH2CH2CI, -CH2CH2CH2C1, -CH2C00C2H5, CH (CH3) COOC2H5 or -CH (CH2C6H5) C00C2H5.

In the preferred compounds according to the present invention, Y-CH2CH2C1 and R is -CH2CH2C1, -CH2CH = CH2 or -CH3. R is more preferably CH2CH2C1, -CH3. The alkyl substituent of 1 to 7 carbon atoms is preferably selected from the group consisting of methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, t-butyl, n-pentyl, isopentyl, n-hexyl , isohexyl and substituted hexyl. The compounds according to the present invention are produced by synthetic methods which are readily known to those skilled in the art which include the chemical synthetic methods described. The present invention also relates to pharmaceutical compositions comprising an antineoplastic effective amount of the compound of 2-ami ocarbonyl-l, 2-bis (methylsulfonyl) -1- (substituted) hydrazine as set forth above.

These pharmaceutical compositions preferably include a pharmaceutically acceptable additive, carrier or excipient. The present invention relates to a method for treating mammalian neoplasia comprising administering an antineoplastic effective amount of the compound of 2-aminocarbonyl-1,2-bis (methylsulfonyl) -1- (substituted) hydrazines to a patient suffering from cancer. . The treatment of solid malignancies comprising administering to a patient an antitumor effective amount of 1 or more of these agents is a preferred embodiment of the present invention. The treatment of leukemias, lung carcinomas, melanoma, reticulum cell sarcoma among several other related disease states can also be performed using the compounds of the present invention.

DETAILED DESCRIPTION OF THE INVENTION The term "neoplasia" is used throughout the specification to refer to the pathological process that results in the formation and growth of a cancerous or malignant neoplasm, that is, abnormal tissue that grows by cell proliferation, often faster than normal and continues to grow after stimuli initiate the sensation of new growth. Malignant neoplasms show partial or complete lack of structural organization and functional coordination with normal tissue and in the majority it invades the surrounding tissues, extending to several sites, and will probably recur after the proposed removal and will cause the death of the patient. unless properly treated. As used herein, the term "neoplasia" is used to describe all cancerous disease states and encompasses or encompasses the pathological process associated with hematogenous, ascites, and solid, malignant tumors. The term antineoplastic effective amount is used throughout the specification to describe an amount of the present compounds that is used to treat a patient suffering from a cancerous tumor to prevent further growth of the neoplasms, bringing that growth under control and preferably, it produces a remission of the tumor. The term "therapeutically effective amount" is used throughout the specification to describe that the amount of the compound according to the present invention that is administered to a mammalian patient especially including a human patient, suffering from cancer, to reduce or inhibit the growth or spread of hematogenous, solid ascitic tumor. Preferably, the compounds according to the present invention will result in remission of the hematogenous, ascites or solid, malignant tumor. In the case of solid tumors, the compounds according to the present invention will inhibit further growth of the tumor tissue and decrease the existing tumor. The present invention is directed to 2-aminocarbonyl-l, 2-bis (methylsulfonyl) -1- (substituted) hydrazine compounds of the formula: CH3S02N (Y) N (CONHR) S02CH3 where Y is -CH3 or -CH2CH2C1; and R is alkyl of 1 to 7 carbon atoms, cyclohexyl, methylcyclohexyl, CH2CH = CH2, -CH2CH2C1, -CH2CH2CH2Cl, -CH2COOC2H5, CH (CH3) COOC2H5 or -CH (CH2C6H5) COOC2H5.

In the preferred compounds according to the present invention, Y is -CH2CHC1 and R is -CH2CH2C1, or -CH3. R is more preferably -CH2CH2C1 or -CH3.

Where R is alkyl of 1 to 7 carbon atoms, R is selected from the group consisting of methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, t-butyl, n-pentyl, isopentyl, n- hexyl, isohexyl, or substituted hexyl. These compounds, which contain a 2-aminocarbonyl group, exhibit improved activity against a broad spectrum of neoplastic disease states, including, for example, numerous solid tumors. In in vivo screening or screening tests, these agents have exhibited broad spectrum activity against a wide range of neoplastic disease states. In one case, where R is CH2CH2C1, this compound exhibited unexpectedly greater antineoplastic activity than mitomycin C or cyclophosphamide, among the most effective commercial antineoplastic alkylating agents. The present compounds represent prodrug forms of intermediates that are believed to exhibit their activity through the mechanisms of chloromethylation, methylation and / or carbamoylation. The compounds according to the present invention are mainly useful for their antineoplastic activity, including their activity against solid tumors. In addition, these compositions may also find use as intermediates in the chemical synthesis of other useful antineoplastic agents which, in turn, are useful as therapeutic agents for other purposes. The compounds according to the present invention are synthesized by the adaptation of techniques that are well known in the art. The 2-aminocarbonyl-1,2-bis (methylsulfonyl) -1- (substituted) hydrazines (I, Y is -CH3 or -CH2CH2C1) are synthesized by reacting 1,2-bis (methylsulfonyl) -1-methylhydrazine or , 2-bis (methylsulfonyl) -1- (2-chloroethyl) hydrazine with the appropriate isocyanate (where R is of the indicated structure or a related alkyl structure) in dry acetonitrile in the presence of triethylamine as shown below. The synthesis of the isocyanate derivative suitable for use in this reaction scheme is well known in the art and uses normal chemical techniques. CH3S02N (Y) NHS02CH3 + RNCO (where Y is -CH3 or -CH2CH2C1) Net3 CH3CN CH3S 02N (Y) N (CONHR) S 02CH3 I I I I. R = -CH2CH2C 1 IV. R = -CH3 V. R = -CH2CH = CH2 VI. R = -CH2CH2CH2C 1 VI I I. R = -CH (CH3) C00C2H5 IX. R = _CH (CH2C6H5) C00C2H5 X. R = --C2C7 alkyl, cyclohexyl or methylcyclohexyl. After the synthesis, the residue is triturated in general, washed with dilute acid, dried, triturated again and recrystallized from a suitable solvent, for example, ethanol or ethanol / petroleum ether. The modification of the described chemical synthesis methods can be easily done by those skilled in the art in order to provide alternative synthetic routes to the present compounds. The present invention also relates to pharmaceutical compositions comprising a therapeutically effective amount of a compound of 2-aminocarbonyl-l, 2-bis (methylsulfonyl) -1- (substituted) hydrazines as set forth above.

A therapeutically effective amount of one or more of these compounds is that amount that can be used to treat patients suffering from cancer such as a malignant tumor. These pharmaceutical compositions also preferably include a pharmaceutically acceptable additive, a carrier or an excipient. In the pharmaceutical compositions according to the present invention which relate to the treatment of malignant solid tumors, those compositions comprising an amount of one or more 2-aminocarbonyl-1 compounds, 2-bis (methylsulfonyl) -1- (substituted) hydrazines as set forth above effective to inhibit the growth of the treated tumor and in certain cases, to actually minimize the treated tumor. One skilled in the art will recognize that a therapeutically effective amount of the compounds according to the present invention that will be used to treat malignant tumors will vary with the disease state or condition to be treated, its severity, the regimen of the treatment to be used, the desired result (remission, tumor shrinkage in combination with surgical techniques or radiation), the type of administration used to distribute or administer the compounds, the in-drug, the compounds used, as well as the patient ( animal or human) treated.

In the pharmaceutical aspect according to the present invention, one or more compounds according to the present invention are preferentially formulated in admixture in a pharmaceutically acceptable additive, carrier or excipient. In general, it is preferred to administer the pharmaceutical composition in the form that can be administered parenterally (preferably, intravenously), but consideration must be given to other formulations administered intramuscularly, transdermally, buccally, cutaneously, suppository, orally or other route. Of course, one skilled in the art can modify the formulations within the teachings of the specification, to provide numerous formulations for a particular route of administration without reverting to the unstable compositions of the present invention or compromising the therapeutic activity. For example, modification of the present compounds to render them more soluble in water or other vehicle, for example, can be easily achieved by minor modifications (salt formulation, esterification, etc.) that are well within ordinary skill in the art. It is also within the ordinary skill to modify the route of administration and regimen and dose of a particular compound in order to manage the kinetics of the present compounds for a maximum beneficial effect in the patient to be treated. Sustained and / or controlled release forms of the pharmaceutical compositions are also contemplated by the present invention. The present compounds are prodrug forms of reactive intermediates. In certain dosage dosage forms, the present compounds can be modified to other forms of prodrug to take advantage of a particular route in the administration of the active compounds. One skilled in the art will recognize how to readily modify the present compounds to alternative forms of prodrugs to facilitate the distribution of the active compounds to a selected site within the patient. The individual of ordinary skill will also take advantage of the favorable pharmacosyne parameters of the prodrug forms, where applicable, in the distribution of the present compounds to a selected site within the patient to minimize the proposed anti-neoplastic effect of the compound.

The amount of the compound included within the therapeutically active formulations according to the present invention is an amount effective to treat the malignant tumor. In general, a therapeutically effective amount of the compound according to the present invention in the dosage form usually varies from less than about 0.05 mg / kg to about 500 mg / kg of body weight of the patient to be treated, or considerably more , depending on the compound used, the type of tumor to be treated, the ability of the active compound to localize in the tissue to be treated, the route of administration and the pharmacokinetics of the compound in the patient. In the case of treatment of solid tumors, the compound is preferably administered in amounts ranging from about 0.05 mg / kg to about 250 mg / kg or more at one time. This dose range generally produces and effective blood level concentrations of the active compound ranging from about 0.01 to about 500 micrograms per ml of blood in the patient to be treated. The duration of treatment may be for one or more days or may last for several months or considerably longer (years) depending on the condition of the disease treated. Administration of the active compound may vary from continuous intravenous drip (to intramuscular, for several oral administrations per day (eg, QID) and may include parenteral administration, including intravenous and intramuscular, oral, topical, subcutaneous, transdermal (which may include a penetration enhancing agent), oral and with suppository among other administration routes. To prepare the pharmaceutical compositions according to the present invention, a therapeutically effective amount of one or more compounds according to the present invention is preferably intimately mixed with a pharmaceutically acceptable carrier according to conventional pharmaceutical mixing techniques to produce a dose. A carrier can take a variety of forms depending on the form of preparation desired for administration,. for example, parenteral or oral. For parenteral formulations the carrier may comprise sterile water or aqueous sodium chloride solution in combination with other ingredients that aid dispersion, such as ethanol and other pharmaceutically acceptable solvents, including DMSO, among others. Of course, where solutions are to be used and maintained as sterile, the compositions and carriers must also be sterile. Injectable suspensions may also be prepared, in which case suitable liquid carriers, suspending agents and the like may be employed. In the preparation of pharmaceutical compositions in the oral dosage form, any of one or more of the usual pharmaceutical media can be used. Thus, for liquid oral preparations such as suspensions, elixiris, solutions, carriers and carrier additives including water, glycols, oils, alcohols, flavoring agents, preservatives, coloring agents and the like can be used. For solid oral preparations such as powders or tablets, capsules and for solid preparations such as suppositories, suitable carriers and additives may be used including starches, sugar carriers, such as manure, mannitol, lactose and carriers, diluents, granulating agents, Lubricants, binders, disintegrating agents and the like, related. If desired, the tablets or capsules may be coated with enteric coating or with a sustained release coating by standard techniques. The compounds and compositions according to the present invention are used to treat cancer in mammals, including humans. In general, to treat malignancies, the compositions will be administered in the form of parenteral, preferably intravenous, doses in amounts ranging from about 25 micrograms to about 500 mg or more one or four times per day. The present compounds are preferably administered parenterally, but may also be administered in an alternative manner, for example, orally even topically or in the form of a suppository. The compounds according to the present invention can be administered alone or in combination with other agents, especially including other compounds of the present invention. In addition, the administration of one or more compounds according to the present invention with other anti-bioplasmic agents, in combination with chemotherapy, such as anti- etabo litos, hetopóxido, dexorubicina, taxol, vincristina, cyclophos famida or mitomycin C, between numerous others, is contemplated by the present invention. While not limited to theory, it is believed that the compounds according to the present invention mainly induce their therapeutic effect in the treatment of malignant tumors by functioning as combined chloroethylation and carbamoylation agents, without providing hydroxyethylation activity. The present invention is now described, completely by way of illustration, in the following examples. It will be understood by a person skilled in the art that these examples are not in any way limiting and that variations of detail can be made without departing from the spirit and scope of the present invention.

EXAMPLES Experimental Section Synthesis. The melting points were determined in capillary tubes in a Thomas-Hoover melting point apparatus and were not corrected.

The 1E NMR spectra were recorded on a varian EM-390 spectrometer with tetramethylsilane as an internal standard. Elemental analyzes were performed by Baron Consulting Col., Orange, CT and the data were within ± 0.4% of the theoretical values for the 2-aminocarbonyl-1,2-bis (methylsulfonyl) -1- (2-chloroethyl) hydrazines.

EXAMPLE 1 Synthesis of 1,2-bis (methylsulfonyl) -1- (2-chloroethyl) -2- (2-chloroethyl) -aminocarbonylhydrazine (III) To a stirred solution of 1,2-bis (methylsulfonyl) -1- (2-chloroethyl) hydrazine (Shyam, et al., J. Med. Chem., 1990, 33, 2259-2264) (2.5kg, 0.010 mol) and 2-chloroethyl isocyanate (1.2 g, 0.011 mol) in dry acetonitrile (100 mL) triethylamine (1.1 g, 0.011 mol) was added at room temperature. After an additional 10 minutes, the reaction mixture was evaporated to dryness in vacuo. The residue was triturated twice with 15 mL quantities of petroleum ether and the petroleum ether layer was discarded each time. The residue was then taken in ethyl acetate (150 mL) and washed with dilute hydrochloric acid (3 x 15 mL). The ethyl acetate layer was dried over anhydrous magnesium sulfate and filtered. Upon evaporation of the solvent, semi-solid residue was obtained which, on trituration with absolute ethanol, gave a white solid. Recrystallization from ethanol gave 1.5 g (42.5%) of the title compound: m.p. 96-97.5 ° C; NMR XH (acetone-d6) 7.0 (br, 1 H, NH), 3.7-4.2 (m 4H, S02NCH2CH2CL), 3.5-3.7 (m, 4H, C0NHCH2CH2C1), 3.5 and 3.3 (2 s, 6H, 2CH3). Anal. (C7Hi5C12N3? 5S2) C, H, N.

EXAMPLES 2-7 SYNTHESIS OF 2-AMINOCARBONIL-1, 2-BIS (MET I LSULFONIL) - 1- (2-CHLOROETHYL) HYDRAZINES The following compounds are prepared using procedures similar to one described in Example 1 for compound III. 1, 2-Bis (met i lsul foni 1) -1 - (2-chloroethyl) -2-methylaminocarbonylhydrazine (IV) was synthesized according to the method of Example 1. Compound IV was recrystallized from ethanol: yield 42.4%, m.p. 146-147.5 ° C; 1K NMR (acetone-d6) 6.7 (br, 1H, NH), 3.7-4.2 (m, 4H, CH2CH2C1), 3.5 and 3.3 (2 s, 6H, 2CH3), 2.9 (d, 3H, NCH3). Anal. (C6H14C1N305S2) C, H, N. According to the method of Example 1, 2-α-laminocarbonyl 1- 1, 2-bis (methylsulfonyl) -1- (2-chloroethyl) hydrazine (V) was synthesized. Compound V was recrystallized from ethanol: yield of 42.2%; p.f. 105-106 ° C; NMR XH (acetone-d6) 6.9 (br, 1H, NH), 5.6-6.1 (m, 1H, CH = C), 5.4, 5.2 and 5.1 (3 d, 2H, C = CH2), 3.7-4.2 (, 6H, NHCH2 and CH2CH2C1), 3.5 and 3.3 (2s, 6H, 2CHA, Anal. (C8H16C1N305S2) C, H, N. In accordance with the method of Example 1, 1,2-bis (methylsulfonyl- 1- ( 2-Chloroethyl) -2- (3-chloropropyl) aminocarbonylhydrazine (VI) Compound VI was recrystallized from ethanol: yield 35.2%, mp 85-86 ° C; XH NMR (acetone-dy) 6.8 (br. , 1H, NG), 3.5-4.2 (m, 4H, S02NCH2CH2C1), 3.4-3.8 (, 6H, CH2CH2CH2C1), 3.5 and 3.3 (2s, 6H, 2CH3) Anal. (C8H17C12N305S2) C, H, N. + It was synthesized according to the method of Example 1 1,2-bis (methylsulfonyl) -1- (2-chloroethyl) -2- (ethoxycarbonylmethyl) aminocarbonylhydrazine (VII). Compound VI was recrystallized from ethanol: Yield 42.2%; p.f. 121-122 ° C; NMR l H (acetone-d6 7.1 (br, 1H, NH), 3.7-4.4 (m, 8H, 0CH2, NHCH2 and CH2CH2C1), 3.5 and 3.3 (2s, 6H, 2 CH3), 1.2 (t, 3H, CCH3 Anal. (C9H? 8ClN307S2) C, H, N. According to the method of Example 1, 1, 2-bis (met i lsul foni 1) - 1 - (2-chloroethyl) -2- ( 1-ethoxycarbonylethyl) aminocarbonylhydrazine (VIII) Compound VIII was recrystallized from ethanol: Yield 28.0%, mp 111-112 ° C; H-NMR (acetone-dβ) 6.9 (br, 1H, NH), 3.7-4.6 ( m, 7H, 0CH2, NHCH and CH2CH2C1), 3.5 and 3.3 (2s, 6H, 2CH3), 1.4 (d, 3H, CHCH3), 1.2 (t, 3H, CH2CH3) Anal. (C? oH2oClN3? 7S2) C , H, N. The 1,2-bis (methylsulfonyl) -1- (2-chloroethyl) -2- (1-ethoxycarbonyl-2-phenylethyl) aminocarbonylhydrazine (IX) was synthesized according to the method of Example 1. Compound IX was recrystallized from ethanol-petroleum ether: Yield 12-8%, mp 106-107 ° C; XH NMR (acetone-d6) 7.1-7.3 (m, 5H, C6H5), 6. 8 (br, 1H, NH), 4.6 (m, 1H, NHCH), 3.6-4.3 (m, 6H, OCH2 and CH2CH2C1), 3.5 (s, 3H, CH3S02), 3.0-3.3 (s, m, 5H, CH2C6H5, CH3S02), 1.2 (t, 3H, CH2CH3).

Anal. (C16H24C1N307S2) C, H, N.

Compounds of 2-aminocarbonyl-1,2-bis (methylsulfonyl) -1-methyldhydrazine containing the same aminocarbonyl substituents are prepared by analogy by following the synthesis protocols described above.

EXAMPLE 8 ANTITUMOR ACTIVITY Antitumor activity was tested in several cell lines: L1210 leukemia, B16F10 melanoma, M5076 reticulum cell sarcoma, M107 lung carcinone and LX-1 lung carcinoma.

Leukemia test L1210 L1210 leukemia cells were obtained from the Frederick Cancer Research Facility, Division of Cancer Treatment Tumor Repository of the National Cancer Institute, and tissue culture was maintained by serial passage. Every 8 weeks, tumor cells were injected intraperitoneally into five CD2F? 8 to 10 weeks of age and allowed to grow for 7 days. The peritoneal fluid was removed and the suspension was centrifuged for 5 minutes at 16000 g. The supernatant was decanted and 10 5 cells / L IN 10 Ml of RPMI 1640 medium supplemented with 10% fetal bovine serum and 1% L-glutamine were again seeded in culture. To assess the antimioplastic strip, i.p. 0.1 mL of the cell suspension containing 105 L1210 leukemia cells in each of the recipient mice. The test compounds were administered over a wide range of dose levels, beginning 24 hours after tumor implantation, and continuing once daily for 6 consecutive days. All drugs were administered i.p. as a solution of 100% dimethylsulfoxide (DMSO), in a volume not exceeding 0.025 ml). in each experiment, the animals were distributed in groups of five mice of comparable weight and were maintained throughout the course of the experiment in Chow pellets from Laboratorio Purina and Agua ad 1 ibi tum. Animals that have control tumor given comparable vehicle volumes were included in each experiment. The mice were weighed during the course of the experiments, and the percentage change in body weight from the beginning to the end of the therapy was used as an indication of the drug's toxicity; the determination of the sensitivity of the neoplasms to these agents was based on the prolongation of the survival time given by the drug treatments.

Results of the L1210 Test The tumor inhibitory properties of compounds III-IX were determined in initial tests by measuring their effect on the survival time of mice having the L1210 leukemia implanted intraperitoneally (ip); The results of these tests are summarized in Table 1, below. With the exception of compound C, all the synthesized agents produced "cures" defined as implants after the tumor 60 days tumor free (in 100% of the mice that have the leukemia LS1210 in at one or more of the dose levels examined after It is conceivable that compound VI failed to do so alone, because it was not evaluated at daily dose levels greater than 15 mg / kg given for 6 consecutive days. Partial healing of the mice that have tumor at the highest dose level examined Compounds III and IV seem to have much better therapeutic potential than those derived from the amino acid ester, that is, compounds VI, VIII, and IX. , the methylurea derivative (IV) produced a cure rate of 40% that mice that have tumor at 5 mg / kg administered for 6 consecutive days without loss of body weight. 100% of mice having lso L1210 leukemia at 10 and 15 mg / kg x 6 with less than a 6% loss of body weight. The derivative (III) of 2-chloroethylurea, which can be considered as a structural analogue of BSU, cured 80 to 100% of leukemic mice at 10 A 20 mg / kg by 6, even at higher doses examined, 3 ie mg / kg appeared to be highly toxic, as silenced by a loss of 10.4 of body weight. The derivative (V) of arilurea was also more highly effective against this tumor, curing 100% of the mice that received a daily dose of 15 mg / kg given for 6 consecutive days. Amino acid ester derivatives, (VII-IX), in general, appear to be considerably less potent than compounds III-V that require daily dose levels in the range of 25 to 100 mg / kg to achieve optimal cure rates , and the early deaths of treated mice occurred at higher doses in each case.

Table 1 Effects of 2-aminocarbonyl-l, 2-bis (methylsulfonyl) -1- (2-chloroethyl) hydrazine. In the survival time of mice that have L1210 leukemia Compound Dosage of% of T / Cc in the Surviving Daily change of 60 days mg / kga Average Weight III 10 -4.7 100 15 4.0 216 80 20 -10.4 239 80 IV 85 + 9.9 234 40 10 - 5.6 100 15 - 2.1 100 Table 1 (Continued) Compound Two is% of T / C in the Survival Daily change 60 days g / kg 'Weight Average V 5 - 1.6 184 20 10 - 2.1 394 40 15 - 2.9 100 VI 5 - 2.8 111 10 - 5.6 187 185 - 8.8 192 20 VII 5 - 1.5 151 10 - 1.4 202 20 15 - 0.5 202 20 - 3.7 191 20 25 - 0.5 100 50 - 2.5 138 80 75 - 1.5 119 60 100 - 4.0 115 Table 1 (Continued) Compound Dosage of T / Cc in the Daily Survival Day of the 60th day mg / kg Weight P romed.iob VIII 5 - 2.0 170 10 - 1.4 178 20 15 - 0.5 185 20 -1.5 227 25 -2.5 239 60 50 -3.3 100 IX 75 - 2.5 125 80 100 - 2.5 118 - . 5 - 2.5 147 10 - 0.5 165 15 - 1.9 160 20 - 1.9 174 25 - 0.9 177 50 - 2.0 225 60 100 - 2.0 100 150 - 5.0 169 20 a Administered ip once daily for six consecutive days, starting 24 hours after the implantation of the tumor, with 5-10 mice that were used per group. B Exchange in average percent in body weight from the beginning of the termination of therapy. % T / C = average survival time for those treated in the control beam x 100; Cures (survivors> of day 60) are listed separately and are not included in this calculation.

Melanoma B16F10 test, Cell Sarcoma of Reticulum M5086, M109 lung carcinoma and LX-1 lung cancer.

The B16F10 melamoma cells were cultured in vitro as in monolayers in minimal essential medium with Hanks salts supplemented with 10% fetal bovine serum and 1% 200 mM L-glutamine solution. Solid tumors were produced in female C57BL / 6 mice from 12 to 14 weeks of age by intradermal injection into the right flank of each mouse of 0.1 mL of a cell suspension containing 10 6 cells B16F10 / mL of freshly treated tripcin cultures. After 10-12 days, animals having tumors of approximately 100 mm3 were treated ip with compound III or IV dissolved in 100% DMSO for 6 consecutive days, and tumor volumes were measured on alternate days until reaching 1000 mm3. Reticulum cell sarcoma M5076 was passed every two weeks by transfer of C57BL / 6 tumor fragments, and lung carcinoma M109 was passed in a similar manner in BALB / c mice. LX-1 human lung carcinoma was passed every two to three weeks in BALB / C-nude antecedent mice (nu / nu). For use in these systems, compound III was dissolved in: (a) 100% DMSO and administered by iv injection in a fixed volume of 10 microliters; or (b) DMSO dissolved with saline at a final concentration of 10% DMSO and administered iv in a volume of 0.01 milliliter / g body weight. These different modes of formulation resulted in differences in the optimal effective dose found in the various tumor systems. Mitomycin C and cyclophosphamide were dissolved and administered in saline. BCNU and MeCCNU were dissolved in ethanol and diluted 1: 9 (v / v) with water before administration. Five mice were used per group of experiments with melanoma B16F10, and 8 mice per group with sarcoma M5076, carcinoma M109 and carcinoma LX-1. A minimum of two dose levels per compound was included in each evaluation and drug therapy was started 24 hours after tumor implantation for M5076 sarcoma and M109 carcinoma. In the LX-1 experiment, the mice having tumor were selected and classified into treatment and control groups on day 6 after tumor implantation such that all tumor weights ranged from 50-100 mg and the average weights of tumor per group were reasonably similar. Therapeutic results were presented in terms of: (a) increases in the time of life reflected by the mean time of relative survival (MST) of the groups treated against the control (ie,% of T / C values) and by long-term survivors, and (b) inhibition of primary tumor growth (ie, TC values) determined when calculating relative mean times for treated (T) and non-treated mice. control treaties (C) to grow tumors of a size of 0.5 g for LX-1 carcinoma or a size of 1 g for murine neoplasms. The tumor weights were interchangeable with the tumor size at the base of 1 mm3 = 1 mg. The activity criterion for the increased life time was T / c of >; 125%. The activity criterion for tumor inhibition was a delay in tumor growth consistent with a logarithmic death cell (LCK). The absolute T-C value needed to achieve this level of efficiency varies from experiment to experiment and depends on the timing of the tumor volume volume of the control mouse in each study. Treated mice that died before day 10 in the M109 ip experiment, or that died before their tumors achieved 0.5 g for LX-1 carcinoma or 1 g in size for all other tumor models were considered to be they have died from the toxicity of the drug. Groups of mice with more than one death due to drug toxicity were reflected in the evaluation of antitumor efficacy. Statistical evaluations of the data were performed using Wilcoxan's generalized test (Gehan, biometrika, 1965, 52, 203-233).

Results of the B16F10 Malanoma Test, Reticulum Cell Sarcoma M5076, M109 Lung Carcinoma and LX-1 Lung Carcinoma Test One of the most active and potent compounds in the series as tested in the L1210 system as described above, compound III, was also evaluated against several other transplanted tumors (Table 2, below). When administered at the high-dose, examined, that is, tre ip dose of 50 mg / kg given at 4-day intervals in the M109 lung carcinoma model implanted by ip this compound produced a% T / C of 267 In the same system, but in the different experiment, the acetyl derivative (X) produced a comparable% T / C of 241 at the highest dose level, examined (60 mg / kg per injection), when the drug was administered ip using the same program (Shyam, et al., J. Med. Chem 1993, 36, 3496-3502). CH3? 02N (CH2CH2C1) N (COCH3) S02CH3 X TABLE 2 Brief description of the optimal antitumor effects of 1,2-bis (meth i sulfo 1) -l- (2-chloroethyl) -2- (2-chloroethyl) aminocarbonylhydrazine (III) in tumors M109, M5076 and LX-1 Tumor, Effective Dosage Schedule% of T / C site Optimum treatment, (Route pathway mg / kg / total injection), and / or tion [T-C, days] M109, iP q4dx3; d.la; 50 a 'c 267 iP M109, se Q4dx3; d.la; 50c a) 115 [8.3] iv 24 [32] c'e b) 143 [9.3] q4dx4; d.la; 64f b) d145 [17.8 IV] M5076, q2dx5; d.la; 48f > 157 (6 / B) iv LX-1, SC q2dx5; d.6a; 40f [14.5] iv a Initial day treatment. Higher dose tested. c administered in 10% DMSO in saline. d Each letter (a, b) means a different experiment. e the dose between brackets that produces the maximum T-C obtained. f administered 100% DMSO. Compound III was also evaluated against lung carcinoma M109 implanted subcutaneously (se). In the initial trial using this model, a dose of 50 mg / kg per injection of this compound was administered intravenously (iv) in 10% DMSO in saline every fourth day for a total of three injections. While the% maximum T / C achieved (115) was not considered to be an active result, a significant delay in tumor growth (T-C) of 8.3 days was observed under these conditions. Mitomycin C, used as a reference drug, produced an effect of maximum% T / C of 103 and a delay in tumor growth of 10 days (data not shown). A subsequent evaluation of compound III was performed using four different doses in a slightly different program, ie, 24, 32, 48 and 64 mg / kg administered every third day for four total injections, and two vehicles, 10% DMSO in solution saline and 100% DMSO. When 10% DMSO was administered in saline, compound III yielded a maximum% T / C of 143 and a maximum delay in tumor growth of 9.3 days at 24 and 32 mg / kg per injection, respectively; the next highest dose evaluated, 48 mg / kg per injection, was excessively lethal. At the highest level evaluated, 64 mg / kg per injection, made possible by the use of 100% DMSO as the vehicle, compound III achieved a maximum% T / C of 145 and a delay in tumor growth of 17.8 days, without causing any of the leanings associated with the treatment. The most recent antitumor effect was statistically superior (p <0.01) to the best T-C value achieved with this compound in 10% DMSO in saline. The cyclophos famida and mitomycin C were induced as reference drugs in this last experiment. The above compound produced a maximum% T / C of 143 and a tumor growth delay of 8.8 days, while mitomycin produced a maximum% T / C of 134 and a CT value of 9.3 days (data not shown) ). As reported above, compound X achieved a maximum% T / C of 136 and a maximum T-C value of 14.5 days against this tumor (Shyam, et al., J. Med. Chem., 1993, 36, 3496). Compound III was also evaluated against reticulum cell sarcoma M5076 implanted. When iv was administered at a level of 48 mg / kg per injection in 100% DMSO every other day for five days, compound III cured 6 of the 8 mice consequently, no mean time (T-C value) reached tumors with a size of 1 gram was expressed for this group. Tumor growth in mice receiving only 100% DMSO was indistinguishable from that of untreated control animals. 1- (2-Chloroethyl) -3- (4-methylcyclohexyl) -1-nitrosourea (MeCCNU) and BCNU were included in this experiment for comparison. The above achieved a maximum of% T / C del28 and a delay in tumor growth from 33.5 days to 16 mg / kg per injection administered iv every fourth day for three total injections, while BCNU, administered iv in the same program treatment, produced a% T / C in excess of 157, with 2 out of 8 cures, and a CT of >; of 62 days (data not shown). Since compound X, a chloroethylating agent without carbamoylation activity, is much less active than compound III or BCNU against this tumor, it seems likely that the generation of an isocyanate intermediate contributes to the anti-bioplasic properties of the compounds. chloroethylation agents against sarcoma M5076. The human lung tumor, LX-1, was grafted onto acrylic mice, was also used to examine the antineoplastic potential of compound III. The treatment was started on day 6 after implantation when the average weight of the tumors was approximately 100 mg. A dose of 40 mg / kg per injection of compound III administered iv in 100% DMSO on a daily schedule for a total of five injections was optimal; this regimen produced an average delay of 14.5 days in the growth of this tumor or an objective size of 0.5 g. This activity level, 1.6 LCK, compared favorably to that obtained with BCNU in the same experiment, which produced a CT of 11.8 days (1.3 LCK) at the optimal dose of 20 mg / kg per injection when administered iv each quarter day for a total of three injections. In addition, both compounds III and IV were evaluated in 100% DMSO against B16F10 melanoma implanted intradermally (id) in mice (Table III). In an initial experiment, compound IV produced a T-C of 15.5 days at a daily dose level of 20 mg / kg administered once daily for six consecutive days beginning on day 10 after implantation. In the same experiment, using the same treatment program, a growth delay of 11 days was obtained with compound III. In the second experiment, when the daily dose of compound IV was reached at 30 mg / kg, a more substantial growth retardation of 25.5 days was achieved while compound III at the same daily dose of 30 mg / kg is less active, with the TC value obtained, which is 13.5 days. In this way, the aminocarbonyl component in this class of agents influenced the magnitude of the antitumor effects obtained against the B16F10 malanoma.

Table 3 Antitumor activity of 1,2-bis (methylsulfonyl) -1- (2-chloroethyl) -2- (2-chloroethyl) aminocarbonylhydrazine (III) and 1,2-bis (methyl sulphonyl) -1- (2-chloroethyl) -2- (2-chloroethyl) -2-methylaminocarbonylhydrazine (IV) against Melanoma B16F10 to treatment in days started. B administered in 100% DMSO. c each letter (a, b) means a different experiment.

Summary In summary, 2-aminocarbonyl-l, 2- bis (methyl sulphonyl) -1- (2-chloroethyl) hydrazines against L1210 leukemia in mice. A representative agent of this class, compound III, was found to have substantial activity in several tumor models from more severe distant sites, which unexpectedly was better than or equal to some of the clinically active alkylating agents, better for comparison in these tests. Additionally, a comparison of compounds III and IV against melanoma B16F10 demonstrated that the aminocarbonyl substituent influenced the degree of neoplastic activity obtainable. It will be understood by those skilled in the art that the foregoing description and the Examples are illustrative in the practice of the present invention, but in no way limiting. The variations of the detail presented herein may be made without departing from the spirit and scope of the present invention as defined by the following claims.

It is noted that in relation to this date, the best method known by the applicant to carry out the present invention, is the conventional one for the manufacture of the objects to which it refers.

Having described the invention as above, the content of the following is claimed as property:

Claims (19)

1. A compound of the formula: CH3S02N (Y) N (CONHR) S02CH3 Characterized because Y is -CH3 or -CH2CH2C1; and R is alkyl of 1 to 7 carbon atoms, cyclohexyl, methylcyclohexyl, -CH2CH = CH2, -CH2CH2C1, -CH2CH2CH2C1, -CH2C00C2H5, -CH (CH3) C00C2H5 O -CH (CH2C6H5) COOC2H5.

2. The compound according to claim 1, characterized in that Y is -CH2CH2C1 and R is -CH2CH2C1, -CH2CH = CH2 or -CH3.

3. The compound according to claim 2, characterized in that R is -CH2CH2C1 or -CH34.

The compound according to claim 2, characterized in that R is CH2CH2C1.

5. The compound according to claim 1, characterized in that the alkyl of 1 to 7 carbon atoms is selected from the group consisting of methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, t-butyl, n -pentyl, isopentyl, n-hexyl, isohexyl or substituted hexyl.

6. A pharmaceutical composition for use in the treatment of malignant tumors in mammals, characterized in that it comprises a therapeutically effective amount of a compound of the formula: CH3S02N (Y) N (CONHR) S02CH3 wherein Y is -CH3 or -CH2CH2C1; and R is alkyl of 1 to 7 carbon atoms, cyclohexyl, methylcyclohexyl, -CH2CH = CH2, -CH2CH2C1, -CH2CH2CH2Cl, -CH2COOC2Hs, CH (CH3) COOC2H5 or -CH (CH2C6H5) COOC2H5.

7. The composition according to claim 6, characterized in that it also includes a pharmaceutically acceptable excipient, additive or carrier.

8. The composition according to claim 6, characterized in that Y is -CH2CH2C1 and R is -CH2CH2C1, -CH2CH = CH2 or -CH3.

9. The composition according to claim 8, characterized in that R is -CH 2 CH 2 Cl or -CH 3.

10. The composition according to claim 9, characterized in that R is -CH 2 CH 2 Cl.

11. The composition according to claim 6, characterized in that the alkyl of 1 to 7 carbon atoms is selected from the group consisting of methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, t-butyl, n -pentyl, isopentyl, n-hexyl, isohexyl and substituted hexyl.

12. The composition according to claim 7, characterized in that it is in the intravenous dose form.

13. The composition according to claim 7, characterized in that it is in the intramuscular dose form.

14. The composition according to claim 7, characterized in that it is in the oral dosage form.

15. Compositions for the use of the treatment of malignant tumors in mammals comprising administering a therapeutically effective amount of a compound of the formula: CH3S02N (Y) N (CONHR) S02CH3 wherein Y is -CH3 or -CH2CH2C1; and R is alkyl of 1 to 7 carbon atoms, cyclohexyl, methylcyclohexyl, -CH2CH = CH2, -CH2CH2C1, -CH2CH2CH2C1, -CH2C00C2H5, CH (CH3) C00C2H5 or -CH (CH2C6H5) COOC2H5.

16. The composition according to claim 15, characterized in that Y is -CH2CH2C1.

17. The composition according to claim 15, characterized in that the alkyl of 1 to 7 carbon atoms is selected from the group consisting of methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, t-butyl, n -pentyl, isopentyl, n-hexyl, isohexyl or substituted hexyl.

18. The composition according to claim 15, characterized in that Y is -CH2CH2C1 and R is -CH2CH2cl, -CH2CH = CH2 or -CH3.

19. The composition according to claim 18, characterized in that R is -CH2CH2C1 or -CH3.