LV11542B - Substituted 1,3,9,11-tetraoxa-4,5,7,8-tetraazaundecadien-4,7-dioxides-5,7, preparation and use thereof - Google Patents

Abstract

The present invention relates to new chemical compounds - substituted 1,3,9,11-tetraoxa-4,5,7,8-tetraazaundecadiene-4,7-dioxides-5,7 with general structure F1, to methods of their preparation, and to pharmaceutical compositions, containing these compounds. Specifically, such compounds can act as potential donors of nitric oxide NO in living organisms, and as strong cytostatic agents they can be employed in cancer chemotherapy.

Description

In particular, these compounds may function as potential donors of nitric oxide NO in organisms and, as potent cytostatic agents, may be used in cancer chemotherapy.

Grounds for the application

Malignancies continue to be one of the leading causes of mortality in most developed countries. It is therefore necessary to develop new, more effective treatments for malignancies, including new treatments. One possible solution here is the use of chemical compounds that are capable of releasing nitric oxide NO, a unique cytotoxic biomediator in the body (Hibbs JB et al., 1988, Biochem. Biophys. Res. Comm. 157, no. ., 1, pp. 87-94.).

From the closest structural analogues of F1 compounds in the scientific and patent literature, nitric oxide NO and nucleophilic adducts of general formula F2 have been proposed as nitric oxide NO donors, cytostatic agents and compounds with a broad spectrum of pharmacological activity, wherein .1 is a radical attached to the active moiety through a heteroatom. , but M is metal and so on. c. cation (Maragos C.M. et al., WO

O

N> 0

X \ X

X N

M +

F2

93/20806, publ. October 28, 1993). Chemically, they are the products of NO addition of nitric oxide to primary and secondary amines, etc. etc.

The other group of known related compounds are alkylation products of the compounds represented by the formula F2, substituted N-oxides of alkoxydialkyltriazenes of the general formula

F3, in which R, - R ₃ are different organic radicals (Keefer LK et. Al., WO 93/07114, publ. 15.04.1993.).

^R 2

N / \ R ₁ N

F3

Compounds of similar structure to 1,7-dioxa-2,3,5,6tetraazaheptadiene-2,5-dioxides-3,5 (see F4)

F4 are known as chemicals (RHflOBCKiiii Β. Η. M flp.,> K. Opr. Xmm., 1980, τ. 16, Bbin. 5, οτρ. 933-936.), But to the best of our knowledge , biomedical information about them has not been published. For the analogues described (see F4) R 1 = R ₄ and these are the lower alkyls, any further conversion being made to the central carbon atom.

Compounds with the general structure F5 are described (Tsien R. Y. et al., WO 94/27957) as photolytically activated nitric oxide NO donors.

F5

In this case, A is a nitrogen or oxygen substituent and B is a photolytic labile group.

The compounds of the general formulas F2 and F3 have several significant disadvantages:

1. The decomposition of compounds F2 and F3 into one of the decomposition products results in the formation of potent carcinogens, N-nitrosamines, as confirmed by the authors themselves (Saavedra JE et al., J. Org. Chem. 1992, vol.57, no. .23., Pp. 6134-6138.).

2. The stability of the compounds F2 and F3 is insufficient for obtaining a medicinal product. The half-lives of the adducts F2 described in the literature in buffer solution at pH = 7.4 and 37 ° C temp. ranging from a few minutes to half an hour (Keei'er, L.K. et al., J. Med. Chem. 1991, vol. 34, no. 11, pp. 3242-3247). This is in line with the authors' idea of direct NO donors, but significantly complicates the use of these agents and their galenical forms for medical use.

To the best of our knowledge, no biomedical information has been published on compounds of general formula F4.

For compounds of general structure F5, a photolytic labile group is provided as a substitute for B (see F5), which significantly limits their potential medical use.

To address the above disadvantages, we provide substituted 1,3,9,1 l-tetraoxa-4,5,7,8tetraazaundecadiene-4,7-dioxides-5,7 as anticancer agents and potential nitric oxide NO donors (see Fl ). Their advantages over the known F2 and F3 are as follows:

1. The possibility of formation of N-nitrosamine in the decay of the compound itself is ruled out, since the molecules F1 do not contain any of the three interconnected nitrogen atoms.

2. For our compounds, the half-life of Fl in buffer solution at pH = 7.4 and 37 ° C. greater than 24 hours, which is sufficient for infusion.

3. Compared to F2 and F3, compounds of the general formula F1 have additional modification possibilities. In particular, the choice of the substituents R [-R ₃ and R ₆ -R _g (see Structure F1) can render the molecule asymmetric with respect to the central carbon atom, allowing for a broader variation in the biological activity of the compounds thus obtained.

Compounds F2 have known cytostatic activity (WO 93/20806); A-375-C6 man. in melanoma cells, they inhibit ³ H-thymidine incorporation in DNA by 50% at 24 - 280 μΜ.

In our studies of the structure-activity regularities of diazene oxides, compounds with general structure F1 exhibit unexpectedly high cytostatic activity. Its compound with R 1 = R ₈ = methyl, R ₅ = ethyl, R ₂ = R ₃ = R ₄ = R ₆ = R ₇ = H (see structure Fl) at a concentration of 4 μΜ shows 27% inhibition of ³ H-thymidine incorporation. DNA of murine lymphocytic leukemia P-388 cells. Reference substance N-methyl-N-nitrosourea this model concentration 1 μΜ showed no ^J H-thymidine activation inhibition.

The results obtained together with the aforementioned advantages of Fl allow the compounds of the general formula Fl to be considered as promising anti-cancer agents.

The physico-chemical properties of the claimed compounds Fl (see experimental part) allow a wide variety of applications to obtain the most appropriate therapeutic forms. Oral dosage forms include:

Solutions containing compounds F1 in therapeutically effective amounts in suitable solvents such as water, ethanol, aqueous ethanol, dimethylsulfoxide, oils, and the like. etc., to which other pharmacologically significant components may be added, as well as stabilizers and preservatives.

2. Tablets, capsules or powders containing the compounds F 1 in therapeutically effective amounts. Various fillers, dyes, preservatives, etc. are acceptable in these forms. etc. the use of pharmaceutical form modifying components such as microcrystalline cellulose, starch, sucrose, lactose, mannitol, gelatin, silica gel, as well as combination with other pharmacologically active components.

3. Aerosols containing therapeutically effective amounts of compounds F1 sprayed with a nebulizer or compressed gas such as freon, propane, etc. etc.

Formulations for parenteral administration include sterile injectable solutions of F1 in aqueous / non-aqueous solvents containing a therapeutically effective amount, as well as microcrystalline suspensions of F1 compounds which may additionally contain preservatives, bacteriostatic agents, buffers, antioxidants, salts, and other pharmaceutical ingredients. for the preparation of preparations. Also included are solutions and suspensions for injection prepared ex tempore.

Experimental part

1. Example.

1,1-dimethyl-1,3,9,1-tetraoxa-4,5,7,8-tetraazaundecadiene-4,7-dioxide-5,7. (A) In the formula Fl R ₁ = R _s = methyl; R ₂ = R ₃ = R ₄ = R ₅ = R ₆ = R ₇ = H.

A saturated solution of EtONa in absolute EtOH is purged with argon, then gaseous NO is bubbled through for several hours until no more precipitate is formed. The resulting precipitate is filtered off, washed with absolute EtOH and dried.

2.0 g of the dry intermediate thus obtained are suspended in 50 ml of acetonitrile, 5 ml of methoxymethyl chloride are added and the suspension is stirred for 6 hours at room temperature. The mixture is then filtered and the filtrate is immersed in a vacuum. The remaining oil crystallizes. The product is recrystallized several times from methyl ethyl ketone-heptane 1/1.

0.25 g of colorless crystals are obtained.

PMR (CDC1 ₃ -TMS):

3.49 (s, 6H, CH ₃ -O) 5.36 (s, 4H, O-CH ₂ -O) 5.89 (s, 2H, N-CH ₂ -N)

Item. analysis

Ο, ΗηΝ, Ο, calc. % N 24.99 Found N 24.73

M = 224.175 C 26.79 H 5.40

C, 26.91; H, 5.41

2. Example.

1,1'-dimethyl-6-ethyl-1,3,9,1-tetraoxa-4,5,7,8-tetraazaundecadiene-4,7-dioxide-5,7. (B)

In the formula F ₁ R ₁ = R _g = methyl; R ₅ = ethyl; R ₂ = R ₃ = R ₄ = R ₀ = R ₇ = H

A saturated solution of sodium n-butylate in n-butanol is purged with argon, then gaseous NO is bubbled through for several hours until no more precipitate is formed. The resulting precipitate is filtered off, washed with absolute n-butanol and dried.

2.0 g of the dry intermediate thus obtained are suspended in 50 ml of acetonitrile, 5 ml of methoxymethyl chloride are added and the suspension is stirred for 6 hours at room temperature. The mixture is then filtered and the filtrate is immersed in a vacuum. The resulting oil does not crystallize, so add a few ml of heptane-CHCl ₃ 1: 1 and leave at -18 ° C. The next day the crystals are filtered. 0.2g.

The product is finally purified by crystallization from absol. EtOH.

Colorless crystals, incl. kuš. 41 ° C.

PMR (CDCl ₃ -TMS) 1.07 (t, 3H, CH ₃ -C)

2:51 (m, 2H, C-CH ₂ -C)

3.51 (s, 6H, CH ₃ -O-)

5.35 (s, 4H, O-CH ₂ -O) 5.93 (t, 1H, CH central)

Item. analysis

Est. % Found

C ₇ H ₁₆ N ₄ O ₆

N, 22.21 N, 21.95

M = 252.229 C, 33.33; H, 6.39

C, 33.34; H, 6.35

Example 3. Cytostatic activity.

Mouse lymphocytic leukemia P-388 cells, mouse line DBA / 2.Cell suspension was prepared in RPMI-1640 medium with 5% serum concentration 10 ⁵ cells / ml. Incubation with preparations 2 h, incubation with Ή-thymidine (lpCi / ml medium) - 1 h.

conc (M) inhibition% A 4.5 x 10 ' ⁴ 89 4.5 x 10 ' ⁶ 0 B 4x 10 ' ⁴ 48 4x 10 ' ⁶ 27th NMU 10 ' ⁴ 35 10 ' ⁶ 0

NMU = N-methyl-N-nitrosourea

References:

1. Hibbs J. B. et. al., Biochem. Biophys. Res. Comm., 1988, vol. 157, no. 1, p. 87-94.

2. Maragos C.M. et. al., WO 93/20806, Publ. October 28, 1993

3. Keefer L. K. et al. al., WO 93/07114, Publ. 4/15/1993

4. RHflOBCKHiŽi Β. H. n flp., X. Opr. Xmm., 1980., τ. 16., βηπ. 5., οτρ. 933936.

5. Tsien R. Y. et. al., WO 94/27957

6. Saavedra J.E. et. al., J. Org. Chem., 1992, vol.57, no. 23, p. 6134-6138.

7. Keefer L. K. et. al., J. Med. Chem., 1991, vol. 34, no. 11, p. 3242-3247.

Claims (8)

PATENT FORMULA We recommend: 1. Chemical compounds substituted by 1,3,9,1-l-tetraoxa-4,5,7,8-tetraazaundecadiene-4,7-dioxides-5,7 of the general formula F1: 2. Pharmaceutical preparations for human and veterinary medicine containing The therapeutically effective amount of the chemical compounds referred to in paragraph 1. 3 1. The appropriate chemical compounds in which the group R _r R ₈ is H or lower alkyl C1-C4. 4. Pharmaceutical preparations for human and veterinary medicine containing The therapeutically effective amount of the chemical compounds referred to in paragraph 3. 5. The chemical compound of claim 3, wherein R 1 and R ₈ are methyl, R ₂ , R ₃ , R ₆ and R ₇ are H and R ₄ and R ₅ are H or lower alkyl ClC 4. 6. Pharmaceutical preparations for human and veterinary medicine containing a therapeutically effective amount of the chemical compounds referred to in point 5. 7. A compound according to claim 5 wherein R ₄ = H and R ₅ is H or ethyl. 8. Pharmaceutical preparations for human and veterinary medicine containing a therapeutically effective amount of the chemical compounds referred to in point 7.