BRPI1010493A2 - antineoplastic pharmaceutical compositions containing substituted nitroaromatic compounds - Google Patents

Abstract

ANTINEOPLASTIC PHARMACEUTICAL COMPOSITIONS CONTAINING SUBSTITUTED NITROAROMATIC COMPOUNDS. The present invention describes antineoplastic pharmaceutical compositions comprising substituted nitroaromatic compounds. The compositions may be used in the treatment of early-stage neoplasms or in combination with other antitumor drugs at later stages of the disease.

Description

"ANTINEOPLASTIC PHARMACEUTICAL COMPOSITIONS CONTAINING SUBSTITUTED NITROAROMATIC COMPOUNDS"

The present invention describes antineoplastic pharmaceutical compositions comprising substituted nitroaromatic compounds. The compositions may be used in the treatment of early-stage neoplasms or in combination with other antitumor drugs at later stages of the disease.

Solid tumors, such as lung, colon and breast carcinomas, are the main types of cancer in men. There is considerable evidence that the presence of hypoxic cells present in solid tumors may limit the effectiveness of radiotherapy. These same cells may also be resistant to many of the commercially available chemotherapeutic agents. However, this apparent obstacle can be explored for the design of agents with selective cytotoxicity for hypoxic cells (Cerecetto, H., Gonzalez, M., Lavaggi, ML Development of Hypoxia Selective Cytotoxins for Cancer Treatment: An Update. Med. Chem. 2, 315, 2006. Papadopoulou, V., Bloomer, WD Exploiting hypoxia in solid tumors with DNA-targeted bioreductive drugs Drugs Fut 29, 807, 2004. Wouters1 BG, Weppler, S.A, Koritzinsky, M., Landuyt, W., Nuyts, S., Theys, J., Chiu, RK, Lambin, P. Hypoxia as a target for combined modality treatments (Eur. J. Cancer. 38, 240, 2002).

In 1972, Lin and colleagues hypothesized that hypoxic cell regions could have a greater capacity for reduction than well-oxygenated cell regions. By analogy, hypoxic cells in solid tumors could exist in a microenvironment that would allow reductive processes to occur. Therefore, it was concluded that these characteristics of hypoxia cells could be explored in the development of chemotherapeutic agents, which would only become cytotoxic after metabolic activation. Thereafter, the concept of bioreductive activation of substances in hypoxia cells has been extensively studied (Dai, J., Liu, Y., Zhou, Y., Nagle, DG Hypoxia-selective antitumor agents: norsesterterpene peroxides from the marine sponge. Diacamus Ieyii preferentially suppresses growth of tumor cells under hypoxic conditions J. Nat. Prod. 70, 130, 2007. Lalani1 AS, Alters, SE, Wong1 A., Albertella1 MR, Cleland, JL, Henner1 WD Selective tumor targeting by the hypoxia-activated prodrug AQ4N blocks tumor growth and metastasis in preclinical models of pancreatic cancer Cancer Clin Res 13, 2216, 2007. Yamazakil, Y., Kunimpto, S., Ikeda1 D. Rakicidin A: A Hypoxia-Selective Cytotoxin. Biol. Pharm. Bull. 30, 261, 2007. Anderson1 RF, Shinde1 SS, Hay, MP, Gamage, SA, Denny1 WA Radical properties governing the hypoxia-selective cytotoxicity of 3-amino-1,2,4-benzotriazine antitumor 1 (4-dioxides, Org. Biomol. Chem. 3, 2167, 2005).

The ability of nitroaromatics to act as bioreducible agents is already well established and therefore these compounds can be used as hypoxia cell selectivity prodrugs (Abreu, FC; Ferraz, PAL; Goulart, MOFJ. Braz. Chem. Soe 13, 19, 2002; Hay1, P., et al., J. Med. Chem., 38, 1928, 1995).

The concept of bioreductive activation of substances in hypoxia cells has been extensively studied (Dai, J. et al. J. Nat. Prod. 70, 130, 2007; Lalani, AS et al. Clin. Cancer Res. 13, 2216 Yamazaki Y. et al Hypoxia-Selective Cytotoxin Biol Pharm Buli 30,261 2007 Cerecetto H. Gonzalez M Lavaggi ML Med Chem 2 315 2006 ) and it is noteworthy that currently two substances, tirapazamine and banoxantrone (AQ4N) are in the final stages of clinical screening studies (Novacea. About AQ4N. Available at: http: //www.redorbit.com/news/ health / 1255929 / novaceas_proofofprinciple_st udy_of_aq4n_in_solid_tumors_published_in / index.html Accessed March 28, 2009, Marcu, L.; Olver, I. Curr. Clin. Pharmacol. 1, 71, 2006). Antitumor activity for hypoxic cells of nitrogen mustards derived from 2,5-dinitrobenzamide is also under investigation, with promising results (Atwell, GJ et al. J. Med. Chem. 50, 1197, 2007). bromomethyl-3-nitrobenzoic is widely used as a substrate in syntheses. Zhang et al described the use of 4-bromomethyl-3-nitrobenzoic acid as a key precursor of benzodiazepine-2-3-dione through a four step sequence including nucleophilic displacement, acylation, simultaneous cyclization reduction and alkylation (Zhang Jinfang; Lou Boliang; Saneii Hossain Application of polymer-bound 4- (bromomethyl) -3-nitrobenzoic acid for synthesis of trisubstituted 1,4-benzodiazepine-2,3-diones Molecular Diversity 6, 13, 2003.

Sun et al reported solid phase synthesis of 3,4-hydro-2 (1 H) -

quinazolinones and 3,4-dihydro-1H-quinazoline-2-thione from Rink resin, acylation of 4-bromomethyl-3-nitrobenzoic acid and amination with primary amines, tin chloride reduction and cyclization (Sun, Q .; Zhou, X.; Kyle, DJ Solid-phase synthesis of 3,4-dihydro-2 (1H) -quinazolinones and 3,4-yl dihydro-1H-quinazolin-2-thiones (Tetrahedron Lett. 42, 4119, 2001) .

Additionally, Oliveira et al evaluated the activity of aromatic nitrocomposites against Trypanossama cruzi, including the trypanocidal activity of 4-bromomethyl-3-nitrobenzoic acid (RB Oliveira, APF Passos, RO Alves, AJ Romanha, MAF Prado, J. Dias de Souza Filho and RJ Alves, Mem. Inst. Oswaldo Cruz, 98, 141, 2003).

Currently we find some patents available for the invention:

EP 866709 discloses a parenteral pharmaceutical composition containing tirapazamine for treating cancer, especially solid tumors, used alone or in combination with radiotherapy or other chemotherapeutic agents.

Patent application W02008118150 discloses a method for treating, preventing or ameliorating hyperproliferative disorders by determining the level of nitric oxide synthase in body fluids and the subsequent administration of bioreducible substances, including banoxantrone. Treatment of cancer patients using hypoxic cell-selective bioreducible substances is under investigation, but these substances are not yet commercially available. New options for substances showing selectivity for hypoxia tumor cells are important in an attempt to overcome the disadvantages of currently investigated options such as normal cell toxicity, inadequate physicochemical properties, and the need for association with classic antitumor drugs.

BRIEF DESCRIPTION OF THE FIGURES

Figure 1: Structural formula of substituted nitroaromatic compounds Figure 2: 4-Bromomethyl-3-nitrobenzoic acid (ANB) structure

Figure 3: DSB (Differential Exploratory Calorimetry) Curve of Pure ANB Figure 4: DSC (Differential Exploratory Calorimetry) Curve of the ANB: β-CD Complex.

Figure 5: DSC (Differential Exploratory Calorimetry) curve of the physical mixture ANB: ΗΡ-β-CD

Figure 6: ΗΡ-β-CD DSC (Differential Exploratory Differential Calorimetry) Curve Figure 7: ΑΒΝ Solubility Diagram: ΗΡ-β-Οϋ

Figure 8: Comparison graph of tumor growth between control group and ANB at a dose of 50 mg / kg (n = 5) and initial tumor volume ~ 250 mm3. Figure 9: Comparison graph of tumor growth between control group and ANB at a dose of 50 mg / kg (n = 6) and initial tumor volume ~ 400 mm3. Figure 10: Comparison chart of tumor growth between control group and ANB complex: ΗΡ-β-CD at a dose of 50 mg / kg (n = 4) and initial tumor volume ~ 400 mm3.

DETAILED DESCRIPTION OF TECHNOLOGY

The present invention describes substituted nitroaromatic containing antineoplastic pharmaceutical compositions having the structural formula of Figure 1.

The substituent "X" of Figure 1 being selected from the group comprising COOH 1 SO 3 H, tetrazoyl, CHO, CH 3, CH 2 OH, CN, COOR 1 CONHR, SONHR 1 NHSO 2 R, NHCOOR, where R may be H, alkyl (C-2 to C-30, with or without branching); aryl (aromatic or heteroaromatic); alkyl aryl (C-2 to C-30, with or without branching, aromatic or heteroaromatic).

And the "Y" substituent of Figure 1 is selected from the group comprising H, F, Cl, Br, I, OH, N3, OPO (OR) 2, NHR, NR2, NR3, OSO2R, OSO2Ar, OAr, OCON1 SH1 SR , SAr; where R may be H, (C-2 to C-30 alkyl, with or without branching); aryl (aromatic or heteroaromatic); alkyl aryl (C-2 to C-30, with or without branching, aromatic or heteroaromatic).

The compositions of the present invention are characterized by the use of substituted nitroaromatic combined with pharmaceutically acceptable excipients. Standard compositions may be liquid, solid or semi-solid. The liquid preparations may be in solution, suspension, emulsion, parenteral or oral form. Semisolids in the form of gels, ointments, creams or pastes and solids in the form of capsules, tablets, dragees or lozenges.

Examples of excipients include methylcellulose, hydroxypropylcellulose, hydroxyethylcellulose, carboxymethylcellulose, polymers derived from acrylic and methacrylic acid, polyethylene glycols, solid vaseline, solid paraffin, lanolin, vegetable oils, mineral oil, cetyl alcohol, sterile alcohol, cetostearyl alcohol, glyceryl monostearate, wax of cetyl esters, nonionic and anionic self-emulsifying wax and sodium lauryl sulfate for semi-solid pharmaceutical forms.

Disintegrating binders1, diluents, lubricants, surfactants such as cellulose, lactose, starch, mannitol, magnesium stearate, talc, colloidal silicon dioxide, magnesium oxide and kaolin for solid preparations.

For liquid dosage forms solubilizers and surfactants such as glycerine, propylene glycol, sucrose, sodium lauryl sulfate and polysorbates may be used. For injections, water for injections may be used. Excipients may also contain minor amounts of additives such as isotonicity and chemical stability enhancing substances such as preservatives, chelators and stabilizers, examples of such substances include phosphate buffer, bicarbonate buffer and Tris buffer, thimerosal, m- or o-cresol, formalin, alcohol benzyl, parabens, EDTA1 BHA, BHT; in addition to sweeteners, colorings and flavorings. Such compositions may be administered intramuscularly,

intravenous, topical, oral, inhalation or as devices that may be implanted or injected. The compositions may be used in the treatment of early-stage neoplasms or in combination with drugs with established antitumor activity at later stages of the disease.

The present invention may be better understood by the following non-limiting examples of the technology:

Example 1: Preparation and Characterization of ANB Inclusion Complexes: HP-β-CD

A) Preparation of inclusion complexes

The ANB: HP-p-CD inclusion complex was prepared by mixing HP-β-CD and ANB (Figure 2) in water and acetone at a 1: 1 molar ratio. The mixture was stirred for 2 hours at room temperature. Subsequently, the acetone was evaporated and the resulting mixture was lyophilized. The lyophilized powder was kept in a desiccator under vacuum.

B) Characterization of inclusion complexes by differential exploratory calorimetry (DSC)

The HP-p-CD: ANB 1: 1 inclusion complex, the physical mixture and the

Pure substances were characterized by differential exploratory calorimetry (DSC).

The pure ANB DSC curve (Figure 3) shows an endothermic event at 134.6 ° C corresponding to the ANB fusion peak and the exothermic event at 183.07 ° C corresponding to its degradation product, indicating that The substance is unstable at temperatures above ~ 150 ° C (temperature at which substance begins the degradation process). The DSC curves of the complex (Figure 4) and the physical mixture (Figure 5) were very similar. Figure 6 shows the C-β-CD DSC curve. In addition to the endothermic event at - 42 ° C, related to water loss, only exothermic events above 180 ° C are observed, corresponding to the formation of degradation products. The intensity of these peaks is lower in the DSC curve of the complex (Figure 4), which may indicate greater protection of the substance within the DC cavity. C) Solubility isotherm measurement

The concentration of soluble ANB at different concentrations of ΗΡ-β-CD is shown in Table 1.

Table 1: Concentration of soluble ANB at different concentrations of ΗΡ-β-CD

[ΗΡ-β-CD] mol / L [ANB] mol / L [ANB] soluble mol / L 0.0 0.02 0.00069 0.004 0.02 0.00096 0.008 0.02 0.00127 0.010 0.02 0.00136 0.015 0.02 0.00171 0.02 0.02 0.00192 0.04 0.02 0.00242 0.06 0.02 0.00250 0.08 0.02 0.00238 0.01 0 .02 0.00246

The ANB: HP-p-CD inclusion complex presented type A solubility diagram, ie when the substrate solubility increases with increasing CD concentration (Figure 7). A linear increase in soluble ANB concentration is observed with the increase in ΗΡ-β-CD concentration. This type of diagram is characteristic of forming soluble inclusion complex.

Example 2: Evaluation of ANB's in vivo antitumor activity and its inclusion complex

A) Induction of Ehrlich ascites tumor in mice

For ascites tumor induction, three Swiss mice were used.

females weighing between 25 and 30 grams for each experiment. Ehrlich cells that were conserved in liquid nitrogen were thawed in a water bath at 37 ° C. They were transferred to a previously cleaned Falcon tube and 0.9% saline was slowly and gradually added thereto until the total cell suspension volume was approximately 20 ml. The suspension was immediately centrifuged for 5 minutes at 5 ° C and 3000 RPM speed. The medium in which the cells were conserved was removed as supernatant. The cells were resuspended with the aid of a Pasteur pipette in about 1.5 ml of 0.9% saline and then the cell concentration in the suspension was determined using the Neubauer chamber. The suspension was adequately diluted and 1x106 cells were injected intraperitoneally into each mouse in a total volume of 0.5 ml.

B) Ehrlich solid tumor induction in mice

Initially, 2 χ 106 Ehrlich tumor cells were taken from ascitic tumor mice and implanted in female Swiss mice, weighing 20 to 25 grams subcutaneously, dorsolaterally. After 10 days, the tumors were measured and the samples to be tested were administered intratumorally.

C) Evaluation of antitumor activity in mice

ANB was administered to a group of 11 animals at a dose of 50

mg / kg, solubilized in saline containing PEG 400 (polyethylene glycol) (40%), 5 in tumors with volumes of 250 mm3 and 6 with volumes of 400 mm3. The same number of animals was used in the control group. The 50 mg / kg ANB: HP-p-CD complex was administered to 4 animals and the same number of animals were used as controls.

Doses were administered intratumorally twice a week for 3 weeks. Tumor volumes (mm3) were calculated from measurements of their size (T) and width (L). Tumor volume was then determined using the formula T (L) 2/2 (Viale, M., Vannozzi MO, Merlo, F., Cafaggi, S., Parodi, B., Esposito, M. Cisplatin Combined with the New Cisplatin-Procaine Complex DPR: In Vitro and In Vitro Studies (Eur. J. Cancer 32A, 2327, 1996). The animals had free access to water and feed and were kept in an environment with light cycle control.

Pure ANB showed significant antitumor activity when compared to the control group, as can be seen in the graphs of figures 8 and 9. It was also observed that treatment is more effective when started in animals with a smaller tumor volume (compare figures 8 and 9). 9). Animals treated with ΑΝΒ: ΗΡ-β-00 showed total tumor reduction (Figure 10).

Claims (8)

1. - SUBSTITUTED NITROAROMATICS characterized by the following structural formula: <formula> formula see original document page 11 </formula> - X being selected from the group comprising COOH, SO3H1 tetrazoyl, CHO, CH3, CH2OH1 CN, COOR1 CONHR1 NHSO2R1 NHCOOR1 where R may be H1 alkyl (C-2 to C-30, with or without branching); aryl (aromatic or heteroaromatic); (C-2 to C-30 alkyl aryl, with or without branching, aromatic or heteroaromatic); - Y being selected from the group comprising H, F, Cl, Br, I, OH1 N3, OPO (OR) 2, NHR1NR2lNR3 OSO2R1 OSO2Ar, OAr, OCOR1 30, with or without branch); aryl (aromatic or heteroaromatic); alkyl aryl (C-2 to C-30, with or without branching, aromatic or heteroaromatic). 2. ANTINEOPLASTIC PHARMACEUTICAL COMPOSITION, characterized in that it comprises substituted nitroaromatics having the following structural formula: <formula> formula see original document page 11 </formula> - X being selected from the group comprising COOH1 SO3H, tetrazoil, CHO, CH3iCH2OH, CN, COOR1 CONHR1 SONHR, NHSO 2 R 1 NHCOOR, where R may be H, alkyl (C-2 to C-30, with or without branching); aryl (aromatic or heteroaromatic); (C-2 to C-30 alkyl aryl, with or without branching, aromatic or heteroaromatic); Y being selected from the group comprising H, F, Cl, Br, I, OH1 N3, OPO (OR) 2, NHR1NR2lNR3, OSO2R1 OSO2Ar1 OAr1 OCOR1 SH1 SR, SAr where R may be H, alkyl (C-2 to C -30, with or without branching); aryl (aromatic or heteroaromatic); (C-2 to C-30 alkyl aryl, with or without branching, aromatic or heteroaromatic); and at least one pharmaceutically and physiologically acceptable excipient or adjuvant. 3. ANTINEOPLASTIC PHARMACEUTICAL COMPOSITION according to claim 2, characterized in that the substituted nitroaromatic compounds are used in their free form included or combined with cyclodextrins. 4. ANTINEOPLASTIC PHARMACEUTICAL COMPOSITION according to Claim 3, characterized in that the cyclodextrins are selected from the group comprising a-cyclodextrin, β-cyclodextrin and γ-cyclodextrin and their derivatives. 5. ANTINEOPLASTIC PHARMACEUTICAL COMPOSITION according to Claim 3, characterized in that the cyclodextrin is preferably hydroxypropyl-β-cyclodextrin. 6. ANTINEOPLASTIC PHARMACEUTICAL COMPOSITION according to Claims 1 to 5, characterized in that the substituted nitroaromatic compounds are used individually or in combination with other antineoplastic agents. 7. ANTINEOPLASTIC PHARMACEUTICAL COMPOSITION according to claims 1 to 6, characterized in that it is administered by oral, subcutaneous, intramuscular, intravenous, intraperitoneal, intratumoral, transdermal routes or as devices which may be implanted or injected. 8.- USE OF SUBSTITUTED NITROAROMATIC COMPOUNDS, characterized by being in the preparation of antitumor drugs.