Pharmaceutical compositions, compounds, methods of producing compounds

 

(57) Abstract:

Describes a new pharmaceutical composition for treatment of a tumor in an animal host that contains an effective amount of-L - enantiomer of formula (I), where R1and R2selected from the group consisting of hydrogen, acyl and C1-C18alkyl, f represents H or F, which is at least 95% free of the corresponding D-enantiomer, or its pharmaceutically acceptable salt in a pharmaceutically acceptable carrier. The composition may find use for the treatment of tumors, including cancer, and other conditions with abnormal or undesirable cell proliferation in animals, including humans. 8 C. and 41 C.p. f-crystals, 5 Il., 3 table.

This invention relates to the field of medical chemistry and, in particular, relates to (-)-(2S, 4S)-1-(2-hydroxymethyl-1,3-dioxolane - 4-yl)cytosine (also referred to as (-)-OddC) or its derivative and their use for the treatment of cancer in animals, including humans.

Background of the invention.

The tumor is a condition with unregulated, disorganized proliferation of cells. The tumor is malignant, or cancerous, if it has the properties of invasiveness and metaalunie membrane, which define the boundaries of tissues, thus, often penetrating into the circulatory system of the body. The concept of metastasis refers to the propensity of the tumor to migrate to other parts of the body and form a zone of proliferation removal from the initial occurrence.

Now cancer is the second leading cause of death in the United States. Cancer is diagnosed in more than 8000000 people in the United States, and it is expected that in 1994 will be delivered 1208000 new diagnoses. Every year in this country from this disease die more than 500,000 people.

The nature of cancer at the molecular level is not fully clear. It is known that the effect on the cells of a carcinogen, such as some viruses, certain chemicals or radiation, leads to changes in DNA that inactivate "suppressive" gene or activate the "oncogene". Suppressive genes are genes that regulate growth, which occurs when mutations are no longer able to restrain cell growth. Oncogenes are initially normal genes (so-called proanagen), which occurs when mutations or when conditions change expression become badcat various normal cellular genes can become oncogenes upon the occurrence of genetic changes. Transformed cells differ from normal cells in many ways, including cell morphology, cell interaction, the content of membrane structures, structure of the cell skeleton protein secretion, gene expression and mortality (transformed cells can grow indefinitely).

All the different cells of the body can be transformed into cells of benign or malignant tumors. The most common place of occurrence of the tumor is a lung, followed by optoceramics region, mammary gland, prostate, bladder, pancreas and then the ovary. Other common types of cancer include leukemia, malignant tumors of the Central nervous system, including malignant brain tumor, melanoma, lymphoma, erythromelas, cervical cancer and cancer of the tissues of the head and neck.

Now the cancer is treated with one or a combination of three types of treatment: surgery, radiotherapy and chemotherapy. Surgical intervention includes volumetric excision of the affected tissue. While surgical treatment is sometimes effective in removing tumors localized in certain kalibawang in other areas such as the spine, as well as in the treatment of disseminated neoplastic conditions such as leukemia.

Chemotherapy involves the interruption of replication of cells or cellular metabolism. It often is used as in the treatment of leukemia and cancer of the breast, lung and testis.

There are five main classes of chemotherapy drugs currently used to treat cancer: natural products and their derivatives; anthracyclines; alkylating agents; antiproliferative funds (also called antimetabolites) and hormonal drugs. On chemotherapeutic agents is often referred to as antineoplastics tools.

It is believed that alkylating agents act by alkylation and cross-linking guanine and possibly other bases in DNA, stopping cell division. Typical alkylating agents include ethyleneimine compounds, alkyl sulphates, cisplatin and various nitrosoanatabine. The disadvantage of these compounds is that they affect not only cancer cells but also other cells, divided naturally, such as bone marrow cells, skin, mucous membrane of the gastro - intestinal of trecento or compounds, which intervene in the process of replication, translation and transcription of nucleic acids otherwise.

It was found that several synthetic nucleosides exhibit anticancer activity. Well-known nucleoside derivative with strong anticancer activity is 5-fluorouracil. 5-Fluorouracil used in the clinic for the treatment of malignant tumors, including, for example, carcinoma, sarcoma, skin cancer, cancer of the digestive organs, and breast cancer. However, 5-fluorouracil causes serious adverse reactions such as nausea, alopecia, diarrhea, stomatitis, leukocyte thrombocytopenia, anorexia, pigmentation and puffiness. Derivatives of 5-fluorouracil with anticancer activity are described in U.S. Patent N 4336381 and in Japanese patent publications NN 50-50383, 50-50384, 50-64281, 51-146482 and 53-84981.

In U.S. Patent N 4000137 described that the product of peroxidation of inosine, adenosine or cytidine with methanol or ethanol has activity against lymphocytic leukemia.

Containerised (which is also referred to as Cytarabine, AGAS and Cytosar) is a nucleoside analogue of deoxycytidine, which was first synthesized in 1950 and introduced into clinical medicine in 1963. In the d against acute lymphocytic leukemia and to a lesser extent, applicable in the treatment of chronic myeloid leukemia and non-Jackinsky lymphoma. The main action of AGAS is the inhibition of DNA synthesis in the nucleus. Handschumacher, R. and Cheng, Y., "Purine and Pyrimidine Antimetabolites", Cancer Medicine, Chapter XV-1, 3rd edition, edited by J. Holland, et al., Lea and Febigol, publishers.

5-Azacytidine is similar to cytidine, which is used primarily for the treatment of acute myeloid leukemia and myelodysplastic syndrome.

2-Tardenois-5'-phosphate (Fludara, which is also referred to as the FaraA) is one of the most active agents for the treatment of chronic myeloid leukemia. The compound acts by inhibiting DNA synthesis. Treatment of cells F-araA is associated with the concentration of cells on the boundary of G1/S-phase and S-phase; thus, it is a medication specific for the S phase of the cell cycle. The inclusion of the active metabolite, F-agaath inhibits the elongation of the DNA chain. F-araA is also a potent inhibitor ribonucleotides, the key enzyme responsible for the formation of dATP.

2-Chloromethoxypropyl used for the treatment of B - cell neoplasms with a low degree of differentiation, such as leukemia, non-Jackinsky lymphoma and hairy cell Le is for the repair of DNA in the remaining cells.

Although a number of chemotherapeutic agents discovered and is currently used for cancer treatment, searching for new tools that would be effective and which would show low toxicity to healthy cells.

Thus, the aim of the present invention is to develop compounds that possess anti-tumor and, in particular, anti-cancer activity.

Another objective of the present invention are pharmaceutical compositions for the treatment of cancer.

A further purpose of the present invention is a method of treatment of cancer.

The essence of the invention.

Describes a method and composition for treating tumors and, in particular, cancer in humans and other animal hosts, which is the introduction of an effective amount of (-) -(2S, 4S)-1-(2-hydroxymethyl-1,3-dioxolane-4-yl) cytosine (also referred to as (-)-OddC, L-OddC or (-)-L-OddC), its pharmaceutically acceptable derivative, including the 5' or N4-alkilirovanie or acylated derivative, or its pharmaceutically acceptable salt, optionally in a pharmaceutically acceptable carrier.

In an alternative implementation described here, the connection could the school or unwanted cell proliferation. Examples include skin diseases such as hyperkeratosis (including ichthyosis, keratoderma, planus and psoriasis, warts, including genital warts, and bullous lesions, as well as any state with abnormal or undesirable cell proliferation, which can be cured with methotrexate. The active compounds described herein may also be used to stimulate or encourage abortion.

In the preferred implementation of (-)-(2S, 4S)-1-(2-hydroxymethyl-1,3-dioxolane-4-yl)cytosine provided in the form of the specified enantiomer (L-enantiomer) and, essentially, in the absence of its corresponding enantiomer (i.e., in enantiomerically enriched, including enantiomerically pure form).

It is believed that (-)-(2S, 4S)-1-(2-hydroxymethyl-1, 3-dioxolane-4 - yl)cytosine is the first example L - nucleoside, which exhibits antitumor activity. (-)- (2S, 4S) -1- (2-hydroxymethyl-1, 3-dioxolane-4-yl) cytosine has a structure represented by formula I

< / BR>
It was found that (-)-(2S,4S)-1-(2-hydroxymethyl-1,3 - dioxolane-4-yl)cytosine shows significant activity against cancer cells and has low toxicity to healthy cells of the host. Neoprene egcogi, optoceramics region, breast, prostate, bladder, nasopharynx, pancreas, ovary, leukemia, lymphoma, cancer of the tissues of the head and neck, malignant tumors of the Central nervous system (including malignant brain tumor, cervical carcinoma, melanoma, and liver cell cancer.

In an alternative implementation of the described method and composition for treating tumors and, in particular, cancer or other abnormal or undesirable proliferative conditions in humans and other host animals, that includes introducing an effective amount of a derivative of L-OddC formula:

< / BR>
where R is F, Cl, -CH3, -C(H)=CH2, -Br, -NO2, -CCH, or-CN, and R1represents hydrogen, alkyl, acyl, monophosphate, diphosphate or triphosphate, or its pharmaceutically acceptable derivative is not necessarily in a pharmaceutically acceptable carrier, preferably, in enantiomerically enriched form.

Although the preferred implementation of this invention is the application of the active compounds or their derivatives or their salts in not occurring configuration (L-configuration), the compounds described herein, or their proactices mixture.

Any compound described herein for use in the treatment of tumors, can be administered in combination or alternation with other anticancer pharmaceuticals in order to increase the effectiveness of treatment. Examples include natural products and their derivatives; anthracyclines; alkylating agents; antiproliferative funds (also called antimetabolites) and hormonal drugs. Specifically, the funds include, but are not limited to, nitrogen mustards (connection chloralkali group at the end of the nitrogen group), ethyleneimine compounds, alkyl sulphates, cisplatin, nitrosamine, 5-fluorouracil, citizenoriented, 5-azacytidine, 2-Tardenois-5'-phosphate, 2-chloromethoxypropyl, tamoxifen, actinomycin, amsacrine, bleomycin, carboplatin, carmustine, cyclophosphamide, cyclosporine, daunorubicin, doxorubicin, interleukin, lomustin, mercaptopurine, methotrexate, mitomycin, tioguanin, vinblastine, growth factors, including G-CSF, GM-CSF, and platelet growth factor; adriamycin, WP-16, hydroxyurea, etoposide; -, - and-interferons and vincristine. Methods of administration of effective amounts of these funds easily defined or described, for example, in The Physician''s Desk Reference, latest edition, published by Medical Economics Data Prod is arranged in the prescribed manner to optimize the effectiveness of concomitant or alternating therapy.

A brief description of the drawings.

Fig. 1 shows EID50(-)- OddC and the combination of (-)-OddC + TSU (tetrahydrouridine, inhibitor titidindezaminaza) for cancer cells of the colon. This graph shows the growth inhibition as a percentage of control growth of the concentration (μm). The graph data separately for (-)-OddC presents) and data (-)-OddC + TSU presents

Fig. 2 is a graph showing the growth of the tumor mass for murine carcinoma (Colon 38), treated twice a day (-)- OddC, doses of 25 mg/kg of body weight. This graph shows the growth of the tumor as a percentage of the initial tumor weight by the number of days. Treatment of mice was performed in 1, 2, 3, 4 and 5 days. The graph data for the control (without the introduction of (-)-OddC presents () and data (-)-OddC presents

Fig. 3 shows the survival rates of leukemic mice with P388, which were treated with (-)-OddC. The graph displays the percentage of survival from the days of treatment. Treatment of mice was performed at 1, 2, 3, 4 and 5 day. This graph shows the survival rate of the control (without the introduction of (- )-OddC) is presented ), the survival rates of mice that were injected (-)-OddC 25 mg/kg of body weight twice a day, presents (----), and the survival rates of mice that were injected (-)-OddC 50 mg/kg mA the values of certain lines of cancer cells to the (-)-OddC based IR. The columns are elongated to right, represent the sensitivity of the cell line to the (-)-OddC, exceeding the average sensitivity of all investigated cell lines. Since the scale of the column is logarithmic, column 2 units to the right means that the connection reaches IR on cell lines in a concentration equal to one-hundredth of the average concentration required for other cell lines, and, thus, the cell line is unusually sensitive to (-)-OddC. The columns are elongated to the left, respectively, mean sensitivity lower than the average.

Fig. 5 is a graph showing inhibition of growth of human tumor (-)-OddC. Three to six mice NCr nude subcutaneously vaccinated 2106cells HepG2 or DU-145 on each side. Treatment was started when tumors were in the progressive stage of growth. Medication was administered twice daily from 0 to 4 inclusive days and the tumor size was measured on the indicated days. Curves a and b show drug effects on HepG2 tumor and tumor DU-145, respectively (-O-Control; Ara --C 25 mg/kg, intraperitoneally; -- (-)-OddC, 25 mg/kg; oral; -- (-)-OddC, 25 mg/kg, intraperitoneally). Each data point represents the average FROM 10 tumors on the chart and six tumors for whom no other method and composition for treating tumors and, in particular, cancer in humans or other host animals, that is the introduction of an effective amount of (-)- (2S, 4S) -1- (2-hydroxymethyl-1, 3-dioxolane-4-yl) cytosine-derived compounds, specified hereinafter, including 5-substituted or 5' or N4-alkilirovanie or acylated derivative or its physiologically acceptable salt, optionally in a pharmaceutically acceptable carrier.

On (-) - (2S, 4S) -1- (2-hydroxymethyl-1, 3 - dioxolane-4-yl)cytosine referred to as "L-nucleoside. Since 2 and 5 carbon dioxolane ring are chiral, non-hydrogen substituents (CH2OH or casinowe base, respectively) can be present as CIS (on the same side) or TRANS (on opposite side) with respect to the system dioxolane ring. Thus, the following configurations are four optical isomers (when dioxolane moiety is oriented in the horizontal plane, so that the oxygen at the 3-position is ahead): CIS (with both groups "up there" that corresponds to the configuration of the natural nucleosides, referred to as the "D"-nucleoside), CIS (with both groups "down", which is not found in nature, configuration, and TRANS (when Deputy when C2 "bottom", the Deputy when C5 "upstairs"). It is believed that (-)-(2S, 4S)-1-(2 - hydroxymethyl-1, 3-dioxolane-4-yl) cytosine or a derivative represents the first example of L-nucleoside, which exhibits antitumor activity. This is unexpected in light of the fact that this configuration is L-nucleoside does not exist in nature.

As used here, the term "enantiomerically enriched" refers to a nucleoside composition that includes at least about 95% and preferably about 97%, 98%, 99% or 100% of an individual enantiomer of this nucleoside. In the preferred implementation of (-)-(2S, 4S)-1-(2 - hydroxymethyl-1, 3-dioxolane-4-yl) cytosine or a derivative or salt presented in nucleoside composition that essentially consists of a single enantiomer, i.e., in the form specified enantiomer (L-enantiomer) and, essentially, in the absence of the corresponding D-enantiomer (i.e., in enantiomerically enriched, including enantiomerically pure form).

The active compound can be introduced, so as any derivative, so that the application of the recipient it was capable of directly or indirectly give the original connection (-)-L-OddC or 5-substituted derivative as otherwise determined selem salt (alternative referred to as "physiologically acceptable salts") (-)- OddC, 5-derivatives, as shown above, and 5' and N4- acylated or alkylated derivatives of the active compounds (for which alternative referred to as "physiologically active derivatives"). In one implementation of the acyl group is an ester of carboxylic acid (-C(O)R), in which decarbonising radical of the ether group selected from an unbranched, branched or cyclic alkyl (C1to C18and more usually from C1to C5), alkaryl, aralkyl, alkoxyalkyl, including methoxymethyl, aralkyl, including benzyl, alkyl or alkoxy from C1to C4; sulfonic esters, such as alkyl - or aralkylamines, including methanesulfonyl, mono-, di - or trifosforny ether, trityl or monomethacrylate, substituted benzyl, trialkylsilyl (for example, dimethyl-tert-butylsilane) or diphenylmethylsilane. Aryl group of esters optimally contain a phenyl group.

Specific examples of pharmaceutically acceptable derivatives of L-O-ddC include, but are not limited to:

< / BR>
where R is F, Cl, -CH3, -C(H)=CH2, -CCH, or-CN, -Br, -NO2, a R1and R2independently selected from the group consisting of hydrogen, alkyl and acyl, specifically including, but opentel, amyl, tert-pentyl, 3-methylbutyryl, hydrosylate, 3-chlorobenzoate, cyclopentyl, cyclohexyl, benzoyl, acetyl, pivaloyl, mesilate, propionyl, butyryl, valeryl, Caproic, Caprylic, capric, lauric, myristic, palmitic, stearic, oleic acids and amino acids, including but not limited to, alanyl, valinol, Latini, isoleucine, Proline, phenylalanine, tryptophanyl, methionine, glycinyl, serinol, threonine, cysteine, tyrosine, asparagine, glutamine, aspartyl, glutamyl, lisini, arginine and histidine. In the preferred implementation derived are presented in the form of L-enantiomer and, essentially, in the absence of its corresponding enantiomer (i.e., in enantiomerically enriched, including enantiomerically pure form).

L-OddC or its derivative may be provided in the form of pharmaceutically acceptable salts. As used here, the term "pharmaceutically acceptable salts or complexes" refers to salts or complexes of L-OddC or its derivatives, that retain the desired biological activity of the parent compound and, if manifest, the minimum toxic effects. Non-limiting examples of such salts are (a) a salt of the added acid, education phosphoric acid, nitric acid and the like), and salts formed with organic acids such as acetic acid, tartaric acid, succinic acid, malic acid, ascorbic acid, benzoic acid, tannic acid, amoeba acid, alginic acid, polyglutamine acid, naphthalenesulfonate, naphthalenedisulfonate and polygalacturonase acid; (b) add salt bases formed with polyvalent metal cations such as zinc, calcium, bismuth, barium, magnesium, aluminum, copper, cobalt, Nickel, cadmium and the like, or an organic cation formed from N,N - dibenziletilendiaminom, ammonia or Ethylenediamine; or (C) a combination of (a) and (b); for example, salt tannate zinc and the like.

Modification of the active compounds, especially N4- and 5'-0-positions, can have a strong influence on the solubility, bioavailability and rate of metabolism of the active ingredients, thus providing the ability to control the transport of active ingredients. Further, modifications can affect the anticancer activity of the compounds, in some cases increasing the activity compared to the reference compound. This can be easily estimated by obtaining the derivative and the leaves in this area.

Thus, the present invention includes the following objects of the invention:

(a) (-)-(2S, 4S)-1-(2-hydroxymethyl-1, 3-dioxolane-4-yl) cytosine and its derivatives and salts;

(b) (+)-(2S, 4S)-1-(2-hydroxymethyl-1, 3-dioxolane-4-yl) cytosine and its derivatives and salts;

() (- /+)- (2S, 4S)-1-(2-hydroxymethyl-1, 3-dioxolane-4-yl) cytosine and its derivatives and salts;

(g) (-)-(2S, 4S) -1- (2-hydroxymethyl-1, 3-dioxolane-4-yl) cytosine and its derivatives and salts or its (+)-enantiomer or racemic mixture and their pharmaceutically acceptable derivatives and salts for use in medical treatment, for example, for the treatment or prevention of tumors, including cancerous tumor;

(d) application of (-)-(2S, 4S)-1-(2-hydroxymethyl-1, 3-dioxolane-4-yl)cytosine and its pharmaceutically acceptable derivatives and salts or its (+) -enantiomer or racemic mixtures and their pharmaceutically acceptable derivatives and salts in the production of drugs for the treatment of tumors, including cancerous tumor;

(e) pharmaceutical compositions containing (-)-(2S, 4S)-1-(2-hydroxymethyl-1, 3-dioxolane-4-yl) cytosine and its pharmaceutically acceptable derivatives and salts or its (+)-enantiomer or racemic mixture or pharmaceutically acceptable derivatives and salts together with fosina, which is:

(1) in the interaction do not necessarily protected cytosine with 1,3 - dioxolane of the formula AND

< / BR>
where R1ais hydrogen or a hydroxyl protecting group, including acyl group, a L is a leaving group; and optionally removing any hydroxyl protective group.

(2) in the interaction of compounds of formula IN

< / BR>
(where R1aspecified above) with an agent serving to convert the carbonyl group in the 4-position brazilero ring in the amino group; and remove any remaining protective groups, which gives the desired product;

(C) the method of obtaining (-)- or (+)- enantiomer (2S, 4S)-1-(2-hydroxymethyl-1, 3-dioxolane-4-yl) cytosine, which is that the compound or its derivative (for example, 5'-ether complex) in a mixture of (-) and (+)- enantiomers are subjected to conditions or conduct interaction with reagents (for example, with a suitable enzyme, serving for the separation of enantiomers and, if necessary, transform derived in the original connection. Alternatively, the mixture can be filtered through a column for chiral liquid chromatography, which separates enantiomers of a given type.

(and) the way to obtain (2S, 4S)-1-(2-hydroc is R>
protected casinowin base, which is optionally substituted in the 5-position, using a Lewis acid, which is not racemized product, such as trimethylsilyltriflate.

With regard to method g) (1), hidroxizina group includes protective groups are described in detail below, including acyl (e.g. acetyl), arlacel (for example, benzoyl or substituted benzoyl), trityl or monomethacrylate, benzyl or substituted benzyl, tizamidine silyl, including trialkylsilyl (for example, dimethyl-tert-Boticelli) or diphenylmethylsilane. Casinowe the connection can be optionally protected tizanidine silyl groups. Protective groups can be removed in the traditional way. Leaving group L is one of the leaving groups known in the chemistry of nucleosides, for example, halogen, such as chlorine, fluorine or bromine, Totila, mesilim, triflate, alkoxy, such as methoxy, ethoxy; or acyl, such as acetyl or benzoyl.

Interaction by the method g) (1) may be carried out in an organic solvent (such as 1,2-dichloroethane or acetonitrile) in the presence of a Lewis acid such as SnCl4, titanium chloride or trimethylsilyltriflate.

Connection aimogasta the compounds of formula C.

< / BR>
(where R1adefined above) with a reducing agent, for example, sociallyengaged, followed by treatment with a suitable conventional reagent to produce the desired intermediate product, for example, acylation with a carboxylic acid anhydride, in particular, acetanhydride for acylation gloriously or romirowsky reagents for halogenation, or alkylating agents.

The connection formulas can be obtained by reacting the compounds of formula D, where R1ais H,

< / BR>
D, or E

with HOCH2CO2H at elevated temperature.

The compound of formula E, where R1adenotes R, can be obtained by ozonolysis allyl simple ester or a complex ester having the formula CH2= CH-CH2-OR, or simply diapir or complex diapir 2-butene-1,3-diol having the formula ROCH2-CH=CH-CH2OR, in which R is a protective group, such as alkyl, silyl or acyl group.

As for how W) 2), the compound of formula C can be treated with 1,2,4-triazole in conjunction with 4-chlorophenyltrichlorosilane with the formation of the corresponding 4-(1,2,4-triazoline) connection, which then convert the by-products of formulas b and C can be obtained, for example, by reacting a suitable (optionally protected) with a compound of the formula And method similar to that described in method g) 1). Uracil and cytosine can be supplied by Aldrich Chemical Co., Milwaukee, W1 53233, USA.

Li-OddC or its derivative can be converted into a pharmaceutically acceptable ester by reacting with a suitable etherification agent, for example, galogenangidridy or carboxylic acid anhydride. L-OddC or its pharmaceutically acceptable derivative can be transformed into its pharmaceutically acceptable salt in the usual way, for example, by treatment with a suitable base. Ester or salt can be converted into the original connection, for example, by hydrolysis.

An alternative implementation of the compounds described herein may be used to treat conditions, specifically different from tumors or cancer, which include abnormal or undesirable cell proliferation. Examples include skin diseases such as hyperkeratosis (including ichthyosis, keratoderma, flat lichen planus and psoriasis, warts, including genital warts, and bullous lesions, as well as any abnormal condition can be used to stimulate or encourage abortion.

Thus, the invention also includes (-)-(2S, 4S)-1-(2 - hydroxymethyl-1,3-dioxolane-4-yl)cytosine and its derivatives and salts or its (+)-enantiomer or racemic mixture and their pharmaceutically acceptable derivatives and salts for use in medical therapy, for example, for the treatment or prevention of conditions with abnormal or undesirable cell proliferation; and the use of (-)-(2S, 4S)-1-(2-hydroxymethyl-1, 3 - dioxolane-4-yl)cytosine and its derivatives and salts or its (+)- enantiomer or racemic mixtures and their pharmaceutically acceptable derivatives and salts when getting medicines to treat conditions of abnormal or undesirable cell proliferation.

II. Getting active compounds.

(-)-L-OddC and its derivatives can be obtained as described above, according to the method described in International PCT Publication N WO 92/18517, published on 29 October 1992, according to the method described in Scheme 1 (see the end of the description) and in the working examples 1-7 below, or by any other method known to specialists in this field. These methods or other known methods can be adapted to obtain derivatives of L-OddC, given by way of example.

4subsequent reduction with NaBH4that no allocation was made in isopropylidene derivative (4). Benzoylation to (5), delete, protect up to (6) and oxidation of the diol (6) gave the acid (7). Oxidative decarboxylation of (7) with Pb(OAc)4in dry THF gave the acetate (8), a key intermediate product in good yield. Acetate are condensed with the desired pyrimidine (for example, sililirovany thymine and N-acetylcytosine) in the presence of TMCOTf, which allowed to obtain a mixture, which was separated on a column of silica gel, which gave the individual isomers (9 and 10). Dibenzoylmethane methanolic ammonia gave the desired (-)-OddC (11).

Example 2: Obtaining (-)-1,6-anhydrous- --L- -holophernes(2).

A mixture of L-gulose (1) (33 g, to 0.127 mol) and 0.5 N. HCl (330 ml, 0,165 mol) was heated under reflux for 20 hours. The mixture was cooled and neutralized to pH 6 using resin (Dovex-2, HCO3-form) while bubbling air. The resin was regenerable and the resin was washed with water (500 ml). The combined filtrate was concentrated to dryness and dried in vacuum over night. The residue was purified using a column (height 5 cm, silica gel, porous, CHCl3-CH3HE, 10: 1) gave a pale yellow solid, which was recrystallized from absolute alcohol, which gave a colorless solid (2) [Rf= 0,43 (CHCl3-CH3HE 5:1), and 7.3 g, 35,52%] . The obtained L-gulose (Rf= 0,07, 11 g) again recyclization that gave (2) (5 g, total yield 60%): so pl. 142,5-145oC;1H NMR (DMCO-d6) 3,22-3,68 (m, 4H, H-2, -3, -4, and-6a), 3,83 (L, J6b,6a= to 7.25 Hz, 1H, Hb-6), 4,22 (pseudo t, J5,6 a= br4.61 and 4,18 Hz, H, H-5), to 4.46 (d, J2-OH,2= 6,59 Hz, 1H, 2 - HE is able to exchange with D2O) to 4.62 (d, J3-OH,3= 5,28 Hz, 1H, 3-OH, is able to exchange with D2O) 5,07 (l, J4-OH,4= 4,84 Hz, 1H, 4-HE is able to exchange with D2O) 5,20 (l, J1,2= 2,19 Hz, 1H, H-1). []2D5-50,011 (s, 1,61, CH3IT).

Example 3: Obtaining (-)-(1'S,2S,4S)-4-(1,2-dihydroxyethyl - 1,2-O-isopropylidene)-2-hydroxymethyl)dioxolane (4).

The solution NaIO4(22,36 g, 0.1 mol) in water (300 ml) was added dropwise to a solution of (2) (11.3 g, 0.07 mol) in methanol (350 ml), cooled to 0oC, for 10 minutes. The mixture was mechanically stirred for 15 minutes. To this mixture d the th substance was filtered and the solid washed with methanol (300 ml). The combined filtrate was neutralized with a 0.5 N. HCl (~200 ml) and concentrated to dryness. The residue was dried in vacuum over night. Syrupy residue triturated with methanol-acetone (1:5, 1200 ml), using a mechanical stirrer (5 hours), and white solid (the first) was filtered. The filtrate was concentrated to dryness and the residue was dissolved in acetone (500 ml) and then p-toluensulfonate (6,63 g, 0.035 mol). After stirring for 6 hours the mixture was neutralized with triethylamine, solid (second) was filtered and the filtrate was concentrated to dryness. The residue was dissolved in ethyl acetate (350 ml) and washed with water (50 ml x 2), dried (MgSO4), filtered and evaporated, to obtain raw (4) (3.6 g) as a yellowish syrup. The aqueous layer was concentrated to dryness and dried in vacuum. The obtained solid substance (first and second) combined with the dried substance from the aqueous layer and was recycliable by stirring for 1 hour in 10% methanol-acetone (900 ml) and p-toluensulfonate acid (16 g, 0,084 mol) and got raw (4) (5.6 g). Raw (4) was purified by dry column of silica gel, CH3OH-CHCl3, 1-5%) to obtain (4) [Rf= 0,82 (CHCl3-CH3HE, 10:1), 8.8 g, 61,84%] in the form of a colorless oil.1H NMR= 6,0 Hz, 1H, CH2OH, is able to exchange with D2O) 4,85 (t = 3.96 Hz, 1H, H-2). []2D5- 12,48 (s, 1,11, CHCl3), Anal. est. for C9H16O5: 52,93; H OF 7.90. Doc.: WITH 52,95; H 7,86.

Example 4; Obtaining (+)-(1'S, 2S,4S)-4-(1,2-dihydroxyethyl - 1,2-O-isopropylidene)-2-(O-benzoyloxymethyl) dioxolane (5).

To a solution of (4) (8.5 g, 0,042 mol) in pyridine-CH2Cl2(1:2, 120 ml) at 0oC dropwise added benzoyl chloride (6.5 ml, 0,056 mol) and the temperature was raised to room temperature. After stirring for 2 hours the reaction was suppressed by the addition of methanol (10 ml) and the mixture was concentrated to dryness in vacuo. The residue was dissolved in CH2Cl2(300 ml) and washed with water (100 ml x 2), brine, dried (MgSO4), filtered and evaporated, to obtain a yellowish syrup, which was purified by chromatography on a column of silica gel (EtOAc-hexane 4-30%), and received (5) [Rand=0,45 (hexane-EtOAc, 3: 1), 10.7 g, 83.4% of] as a colourless oil.1H NMR (CDCl3) of 1.35 and 1.44 (2 x s, 2 x 3H, isopropylidene), 3,3-of 4.35 (m, 6H, H-4, -5, -1' -2'), of 4.44 (d, J= 3,96 Hz, 2H, CH2-OBz), from 5.29 (t, J=3,74 Hz, 1H, H-2), 7,3-7,64, 8,02-8,18 (m, 3H, 2H, -OBz). []2D5+ of 10.73 (s, 1,75, CH3IT). Anal. est. for C16H20O6: 62,33; H 6,54. Doc.: WITH 62,39; H 6,54.

The residue was purified by chromatography on a column of silica gel (EtOAc-hexane 10 - 33%), with (6) [Rf=0,15 (hexane-EtOAc, 1:1), to 4.92 g, 99.2 percent] in the form of a colorless syrup.1H NMR (DMSO-d6) of 3.43 (m, 2H, H-2'), 3,67 of 4.1 (m, 4H, H-4, -5 and -1'), 4,32 (d, J=to 3.73 Hz, 2H, CH2-OBz), 4,60 (t, J= 5,72 Hz, 2'-HE is able to exchange with D2O), 5,23 (t, J=3,96 Hz, 1H, H-2), 7,45-7,7, 7,93-8,04 (m, 3H, 2H, -OBz), []2D5+ 9,16 (c, 1,01, CHCl3), Anal. est. for C13H16O6: 58,20; H 6,01. Doc.: WITH 58,02; H 6,04.

Example 6: Obtaining (-)-(2S,4S)- and (2S,4R)-4-acetoxy-2-(O - benzoyloxymethyl)dioxolane (8).

To a solution of (6) (3.04 from g to 0.011 mol) in CCl4:CH3CN (1:1, 160 ml) was added a solution of NaIO4(10,18 g 0,048 mol) in water (120 ml) followed by the addition of hydrate RUO Li2(0.02 g). After the reaction mixture was stirred for 5 hours solid 2CI2(100 ml) and the aqueous layer was extracted with addition of CH2Cl2(100 ml x 2). The combined organic layer was washed brine (50 ml), dried (MgSO4), filtered, evaporated to dryness in a vacuum for 16 hours, to obtain raw (7) (2.6 g, 91%).

To a solution of crude (7) (2.6 g, 0.01 mol) in dry THF (60 ml) was added Pb(SLA)4(5,48 g, 0,0124 mol) and pyridine (0,83 ml, 0,103 mol) in an atmosphere of N2. The mixture was stirred for 45 minutes in an atmosphere of N2and a solid substance was removed by filtration. The solid is washed with ethyl acetate (60 ml) and the combined organic layer evaporated to dryness. The residue was purified by chromatography on a column of silica gel (hexane-EtOAc, 2:1) and received (8) [Rf= to 0.73 and 0.79 (hexane-EtOAc, 2: 1), 1,9 g, 69,34%] in the form of a colorless oil.1H NMR (CDCl3) 1,998, 2,11 (2 x s, 3H, -SLA), 3,93-to 4.33 (m, 2H, H-5), 4,43, 4,48 (2 x d, J=to 3.73, 3,74 Hz, 2H, CH2OBz), 5,46, 5,55 (2 x t, J= 4,18, 3,63 Hz, IH, H-2), 6.42 per (m, 1H, H-4), 7,33-7,59, 8,00-8,15 (m, 3H, 2H, -OBz). []2D5- of 12.53 (s, 1,11, CHCl3). Anal. est. for C13H14O6: 58,64; H 5,30. Doc.: C 58,78; H 5,34.

Example 7: Obtaining (-)-(2S,4S)-1-[2-(benzoyl)-1,3 - dioxolane-4-yl]cytosine (9) and (+)-(2S, 4S)-1-[2-(benzyloxy)-1,3 - dioxolane-4-yl]loa (10).

A mixture of N4-azet the mini quantity) was heated under reflux for 4 hours in nitrogen atmosphere. The obtained clear solution was cooled to room temperature. This Siciliana acetylcytosine solution was added (8) (1.0 g, 3,76 mmol) in dry dichloroethane (10 ml) and f (1,46 ml of 7.55 mmol). The mixture was stirred for 6 hours. Was added a saturated NaHCO3(10 ml) and the mixture was stirred for 15 minutes and filtered through a pad of Silica. The filtrate is evaporated and the solid was dissolved in EtOAc and washed with water and brine, dried, filtered and evaporated, to obtain the crude product. This crude product was purified on a column of silica gel (5% CH3HE/CHCl3) and received clean-mixture (9) and (10) (0.40 g, 30%) and a mixture (13) and (14) (0,48 g, 40%). Compound (14) was again etilirovany to separate, combined, the mixture was divided into long column of silica gel (3% CH3HE/CHCl3) and received (9) (0,414 g, 30.7) and (10) (0,481 g, 35.6 percent) in the form of foam. These foam triturated with CH3HE received a white solid. 9: UV (CH3OH)max298 nm; Anal. (C17H17N3ABOUT8) C, H, N. 10: UV (CH3OH)max298 nm.

Example 8: Obtaining (-)-(2S,4S)-1-(2-hydroxymethyl-1,3 - dioxolane-4-yl)cytosine (11).

The solution (9) (0.29 grams, 0,827 mmol) in CH3HE/NH3(50 ml, saturated at 0oC) was stirred at whom kapelevich plates (20% CH3HE/CHCl3with getting oil. He was led out of CH2Cl2/hexane and was obtained (11) (0,136 g, with 77.7%) as a white solid. UVmax278,0 nm ( 11967) (pH 2), 270,0 nm ( 774) (pH 7), 269,0,0 nm ( 8379) (pH 11; Anal. (C8H11N3ABOUT4) C, H, N).

II. Farmacevtska composition.

People, horses, dogs, cows and other animals, in particular, mammals, suffering from tumors and, in particular cancer, can be treated by injecting the patient an effective amount of (-)-OddC or its derivative or its pharmaceutically acceptable salt, optionally in a pharmaceutically acceptable carrier or diluent, either by itself or in combination with other known anti-cancer pharmaceuticals. This treatment can also be combined with other conventional cancer therapies, such as radiation therapy or surgical treatment.

These compounds can be administered by any appropriate route, e.g. orally, parenterally, intravenously, intradermally, subcutaneously, or topically in the form of a liquid, ointment, gel or solid substance or in the form of an aerosol. The active compound is included in the composition with a pharmaceutically acceptable carrier IPO desired indication, without causing the patient undergoing treatment, serious toxic effects. The preferred dose of a compound for the treatment of all these conditions ranges from about 10 ng/kg to 300 mg/kg, preferably from 0.1 to 100 mg/kg / day, more typically from 0.5 to about 25 mg per kilogram of body weight of the recipient per day. The usual dose for local application will range from 0.01 to 3% by weight in a suitable carrier.

The connection is easy to enter in any suitable standard dosage form, including, but not limited to, a standard dosage form containing from 1 to 3000 mg, preferably from 5 to 500 mg of active ingredient in the standard dosage form. When oral administration is usually convenient dose of 25-250 mg

The active ingredient preferably is introduced in order to obtain the maximum concentration of active compound in the plasma of about 0,00001-30 mm, preferably about 0.1-30 microns. This can be achieved, for example, by intravenous injection of a solution or composition of the active ingredient, optionally in saline solution or in aqueous medium or in the form of a bolus of the active ingredient.

The concentration of the active compounds in addition to the a, as well as other factors known to specialists in this field. It should also be noted that the levels of dosages will also vary depending on the severity of the condition to be relieved. Next note that for each individual subject of a special scheme of administration of a drug must be installed on the lapse of time in accordance with individual need and the professional judgment of the person typing or watching the introduction of the compositions, and that the intervals of the concentrations set forth herein are presented only as examples and are not intended to limit the scope or practice of the claimed composition. The active ingredient may be administered at once or may be divided into a number of smaller doses to be introduction over varying intervals of time.

Compositions for oral administration will generally include an inert diluent or edible carrier. They can be enclosed in gelatin capsules or compressed into tablets. In order therapeutic oral administration of the active compound or its proletarienne derivative may be mixed with excipients and used in the form of tablets, pistillate in the composition as part of the composition.

Tablets, pills, capsules, lozenges and the like can contain any of the following ingredients, or compounds of a similar nature: a binder such as microcrystalline cellulose, resin tragakant or gelatin; an excipient such as starch or lactose, a dispersing agent, such as alginic acid, Primogel, or corn starch; lubricating substance, such as magnesium stearate or Sterotes; a moving substance, such as colloidal silicon dioxide; sweetening agents such as sucrose or saccharin; or corrigent, such as peppermint, methyl salicylate or corrigent with orange. When the standard dosage form is a capsule, it may contain in addition to materials of the above type, a liquid carrier, such as non-volatile oil. In addition, the standard dosage forms can contain various other materials, which change the physical form of the dosage unit, for example, a shell of sugar, shellac or intersolubility substances.

The active compound or its pharmaceutically acceptable salt may be administered as a component of an elixir, suspension, syrup, capsules, chewing gum, and the like. The syrup can krasiteli and corrigentov.

The active compound or its pharmaceutically acceptable salt can also be mixed with other active materials that do not reduce the desired action, or with materials that support the desired action, such as other anti-cancer remedies, antibiotics, antifungal, anti-inflammatory or antiviral compounds.

Solutions or suspensions used for parenteral, intradermal, subcutaneous, or topical application can include the following components: a sterile diluent such as water for injection, saline solution, fixed oils, polyethylene glycols, glycerine, propylene glycol or other synthetic solvents; antibacterial agents such as benzyl alcohol or methylparaben; antioxidants such as ascorbic acid or sodium bisulfite; chelating agents such as ethylenediaminetetraacetic acid; buffers such as acetates, citrates or phosphates and agents for regulating toychest, such as sodium chloride or dextrose. The preparation for parenteral administration can be enclosed in ampoules, disposable syringes or vials containing multiple doses, made of glass or the th saline or phosphate saline buffer (FSB).

In one implementation, the active compounds are prepared with carriers that will protect the compound against rapid withdrawal from the body, for example, in the form of a controlled release formulation, including implants and microencapsulation delivery system. Can be applied amenable to biological degradation, biocompatible polymers, such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen, complex poliorcetes, polylactic acid. Methods of obtaining such compositions will be evident to the experts in this field.

Liposomal suspensions can also serve as pharmaceutically acceptable carriers. They can be obtained by methods known to experts in this field, for example as described in U.S. Patent N 4522811 (which is incorporated herein by reference in full). For example, liposomal formulations can be obtained by dissolving suitable lipid(s) (such as stearoylethanolamine, stearoylethanolamine and cholesterol) in an inorganic solvent that is then evaporated, and the surface of the vessel there is a thin film of dried lipid. An aqueous solution of active compounds contribute then to the vessel. The vessel then the enemy is Zuya, thus, the liposomal suspension.

III. Biological activity.

Among specialists in this field a large number of biological assays used and is generally accepted for the evaluation of anticancer activity of compounds. Each of these methods can be applied to evaluate the activity of compounds described herein. One generally accepted method of evaluation activity is the use of panels for testing lines of cancer cells from the National Cancer Institute ("NCI"). These tests evaluate the anticancer activity of individual compounds in vitro and are of prognostic data in relation to the use of these compounds in vivo. Other tests include assessing the in vivo effect of the compounds on the human or murine tumor cells implanted in mice or grafted to nude mice. (-)-OddC examined for anti-cancer activity in vivo on line leukemia cells P388 and lines of cancer cells from the colon, C38. In examples 9 and 10 presents the details of the experiments and the results of these studies.

Example 9. Treatment of leukemia cells P388 (-)-OddC in vivo.

The BDF1 mice, obtained from Southern Research Institute, Alabama, implanted intraperitoneally 1 is n days after the implantation of tumor cells. Using this Protocol, it was shown that 75 mg/kg/dose is toxic to mice.

In Fig. 3 and table. 1 shows the results of these studies. In Fig. 3 (a) presents data for control animals not subjected to treatment), (----) represents the survival rates of animals that were administered (-)- OddC 25 mg/kg twice a day, and (O) represents the survival rates of mice that were injected (-)-OddC once a day 50 mg/kg of body weight. Of the six mice, which were treated, 25 mg/kg/dose (-) -OddC, one mouse lived a long time, and the life of the remaining five mice increased by 103%.

Example 10. Treatment of tumor cells 38 of the large intestine (-)-OddC in vivo.

Tumor cells 38 of the large intestine implanted subcutaneously to mice BDF1. (-)-OddC was administered to mice twice a day for five days at a dose of 25 mg/kg/dose. The growth of tumor cells from the large intestine was stopped, as shown in Fig. 2. In Fig. 2 (a) presents data for the control animals, and () represents the data on mice, which were treated with (-)- OddC.

Example 11. The study of (-)-OddC in vitro.

(-)-OddC was assessed by the program NCI cancer screening. The test allows to measure the inhibition of various cancer lines cleto>Table. 2 also gives the concentration at which in the tested cell lines was observed IR and feast. IR, PIR and LC50 are values representing the concentrations at which PR (percent inhibition of growth), as defined below, is equal to +50, 0, and -50, respectively. These values were determined by interpolation curves dose-response obtained for each cell line are shown as functions ETC from log10the concentration of (-)-OddC.

PR was measured by the effect of (-)-OddC on the cell line and was calculated by one of the following two expressions:

If (Average ODtest- The average ODtol) 0,

then

PR = 100 (Average ODtest- The average ODtol)/(Mean ODcounter- The average ODtol).

If (Average ODtest- The average ODtol) < 0,

then

PR = 100 (Average ODtest- The average ODtol)/(Mean ODtol).

Where:

The average ODtol= Measured average optical density of the color that gives SRB, right before the impact on the cells of the tested compounds.

The average ODtest= Measured average optical density of the color that gives SRB, after 48 cbradney optical density color which gives SRB, after 48 hours in the absence of effects on cells of the tested compounds.

In table. 2 the first two columns describe the Subpanel (e.g., leukemia) and cell line (e.g., CCRF-CEM), which was treated with (-)-OddC. Column 3 shows log10where there is IR, and column 4 shows log10where there is a feast. If these options are the answer cannot be obtained by interpolation, the value given for each answer option, represents the highest tested concentration, and it is preceded by the icon ">". For example, if all values of IR at all concentrations of (-)-OddC, data for individual cell lines exceed +50, this parameter can not be obtained by interpolation.

Fig. 4 is a graph which shows the relative selectivity (-)-OddC in relation to a specific cell line. The columns are elongated to right, represent the sensitivity of the cell line to the (-)-OddC greater than average sensitivity of all tested cell lines. Since the scale of the column is logarithmic, column 2 units to the right means that the connection reaches IR on cell lines at concentrations equal what I am unusually sensitive to (-)-OddC. The columns are elongated to the left, respectively, mean sensitivity lower than the average. Data cell lines can be easily determined from the table. 2, since the value of log10concentration will be preceded by ">".

In Fig. 4 you can see that at least one cell line from each tested type of cancer cells showed sensitivity to (-) -OddC. Certain cell lines, prostate cancer, leukemia cell lines and cell lines from colon exhibit extreme sensitivity to (-)-OddC.

Example 12. Comparison of (-)-OddC and araC.

As discussed in the Assumptions of the invention, citizenoriented (which is also referred to as Cytarabine, araC and Cytosar) is a nucleoside analogue of deoxycytidine used for the treatment of acute myeloid leukemia. It also shows activity against acute lymphocytic leukemia and, to a lesser extent, applies in the case of chronic myeloid leukemia and non Hodgkin's lymphoma. The main mechanism of action of araC is the suppression of DNA synthesis in the nucleus. It was interesting to compare the toxicity of (-)-OddC and AraC in relation to tumor cells.

Cells in logarithmic growth phase were sown with the PLO maintained for three generations. At the end of this time tested with methylene blue and/or number of cells was counted directly. Methylene blue is a dye that is stoichiometrically binds to proteins of viable cells and can be used for indirect counting the number of cells (Finlay, 1984). IC50 values were determined by interpolation of data on the graph. Each displayed value is the average standard deviation of five experiments, each data point is found in two copies.

In all cell lines tested (-)-OddC was more toxic than AraC. (-)-OddC was significantly more effective than AraC, in the case of cell line KB carcinoma of the nasopharynx and in two lines DU-145 and PC-3 carcinoma of the prostate. The HepG2 cells originate from hepatic cell carcinoma, and the line 2.2.15 comes from HepG2 cells, transfected with a copy of the genome of hepatitis C CEM Cells originate from cells of acute lymphoblastic leukemia. (-)-OddU, the compound that will be formed by the deamination of (-)-OddC, showed no toxicity against any of the tested cell lines. Enzymatic studies show that in contrast to the AraC,s is the substrate desaminase.

It was determined that (-)-OddC can be phosphorylated to the mono-, di - and triphosphorylated in vivo. It turns out that (-) -OddC shows cellular toxicity in phosphorylated form, as the cells are not able to phosphorylation connection, much less sensitive to the compound. The first enzyme responsible for favorilerine is a human deoxycytidylate. Enzymatic in vitro studies show that (-) -OddC can be phosphorylated by this enzyme.

Unlike araC, (-) -OddC is not subject to deamination by titidindezaminazoy. The presence titidindezaminaza in the tissues of solid tumors may be a key contributing factor responsible for the absence of araC activity in solid tumors. This may partly explain why (-)-OddC shows activity against HepG2 cells in nude mice, whereas araC such activity does not show. This also explains why (-)-OddC has a spectrum of antitumor activity than that of araC. In addition, the presence titidindezaminaza in the gastrointestinal tract may play an important role in the fact that araC cannot be taken orally.

Biochemical studies of (-)-OddC.

The AraC cytotoxicity, (-)-OddC and and rats with immune Taconic) were inoculated subcutaneously on each side 2106cells HepG2 or DU-145 and tumors were allowed to grow. Treatment was started when tumors reached 100-250 mg, which was determined by measurement using stangen-compass and was calculated by the formula:

The tumor weight (mg) = length (mm) x width (mm) - 2

Medication was given in the dose in days from 0 to 4 inclusive and the sizes of tumors were measured every few days. Growth curves of the tumors were built as described in Bell et al. Cancer (phila.)/ 36:2437-2440 (1975), and shown in Fig. 5 (a) and 5(b).

Toxicity was evaluated by body weight changes.

Although AraC toxicity in vitro was similar to that of (L)- OddC, AraC has been ineffective in this animal model. Enzymatic analysis of the extract of the tumor showed that it was not due to increased activity DCD or reduced activity DCK, but could be the result of active metabolism AraC in the liver, which has high levels DCD. Unlike AraC (L)- OddC was effective, as in the case of HepG2 xenograft and in the case of xenograft DU-145. The net cell death (log 10), calculated for HepG2 tumors, amounted to 0.67 and 0.87 for intraperitoneal and oral treatment, respectively. Tumor DU-145 decreased in size, with half of them fully A drugs but the growth was again stopped after 47 days. 60 day animals were killed and tumors removed. The tumor was necrotic morphology with a very small number of cells, is able not to include tripney blue. In addition, this tissue was impossible to determine any enzymatic activity. The administered dose AraC and (L)-OddC were toxic equally, indicating that the loss of animal weight, and preliminary experiments on the toxicity suggest that 25 mg/kg twice a day may be the maximum tolerated dose for five days of continuous treatment. It may be preferable such scheme of the experiment, when the medicine is injected intermittently.

Data obtained in vitro and in vivo, shown here, show that (L)-OddC has significant anticancer activity and can in many settings to exceed the currently available analogs of deoxycytidine. He is not only the first L-nucleoside analogue, which is shown having anticancer activity, but is also the first true terminator, is able to suppress tumor growth, although its unnatural stereochemistry does not prevent the activation of (L)-OddC metabolic enzymes or its inclusion in NAM the m relation, he is active in solid tumors, which are usually not receptive to therapy nucleoside analogues. Drug 2', 2'-diverticulitis (gemcitibine), which is currently undergoing clinical trials for the treatment of solid tumors, is still sensitive to inactivation DCD (16). Because increased levels DCD is the mechanism by which cells acquire resistance to the analogues dcid, such as AraC (17), (L)-OddC can be used in the treatment of patients who have become resistant to these drugs.

IV. The use of (-)-OddC in oligonucleotide and antisense technology.

The concept of antisense technology, in General, relates to modulation of gene expression through a process in which synthetic oligonucleotides hybridized with complementary sequence of nucleic acid for inhibiting the transcription or replication (if the sequence of the target is DNA, inhibition of translation (if the sequence of the target is PHK) or inhibition processing (if the sequence of the target is pre-PHK). Using this technology, you can modulate a wide range of cellular activities. A simple example of the two is the first implementation of the synthetic oligonucleotide hybridized with specific gene sequence in double-stranded DNA with the formation of triplex complex (triplex), which inhibits the expression of a given gene sequence. Antisense oligonucleotides can also be used for indirect activation of gene expression by inhibition of biosynthesis of natural repressor or for direct activation by reducing the termination of transcription. Antisense oligonucleotide therapy (AOT) can be used for inhibiting the expression of abnormal genes, including genes that are involved in uncontrolled cell growth is benign or malignant tumors or who are involved in the replication of viruses, including HIV and hepatitis C.

Stability of oligonucleotides against nucleases is an important factor for applications in vivo. It is known that the 3'-and Exo-nuclease activity is responsible for much of the destruction of the unmodified antisense oligonucleotides in serum. Vlassov, V. V., L. A. Yakubov, in Prospects for Antisense Nucleic Acid Therapy of Cancers and AIDS, 1991, 243-266, Viley-Liss, Inc., New York; Nucleic Acids Res., 1993, 21, 145.

Substitution of the nucleotide on the 3'end of the oligonucleotide (-)-OddC or its derivatives may increase the stability of the oligonucleotide against the destruction of the 3'-ectonucleoside. Alternative or in addition, an internal nucleotide can be zames is lsua given a description here, the person skilled in the art will be able to apply (-)-OddC or its derivatives to stabilize a wide range of oligonucleotides against destruction by ectonucleoside and endonucleases, including nucleosides used in antisense oligonucleotide therapy. It is believed that all these implementation fall within the scope of this invention. Example 13 describes one non-limiting, example of the application of (-)-OddC to inhibit the activity of 3'-accoucheuse.

Example 14. Obstruction of the 3'-ectonucleoside activity by (-)-OddC.

Cytosolic ectonucleoside human activity in H9 human T-lymphocytic leukemia cells) was determined by securitysage analysis in the gel. Briefly, the 3' end of the substrate received from the primer DNA length from 20 to 23 bases with sequence 1 (see the end of the description).

The primers were labeled at the 5'-end with [Y32P] ATP, hybridized with complementary matrices PHK and terminali on the 3'-end of dTTP (20 mer), dCTP (23 mer) or (-)-OddCTP (23 mer) in the reaction catalyzed by reverse transcriptase of HIV-1. In these conditions, a 20 mer was terminials with dTMP (A) 23 mer was terminials dCMP (In) or (-)-O-ddCMP (C). Data single-stranded DNA substrates used for is 10 ál, containing 50 mm Tris-HCl, pH 8.0, 1 mm MgCl2, 1 mm dithiothreitol, 0.1 mg/ml bovine serum albumin, 0,18 µci/ml of 3' -terminal substrate and 2 ál ectonucleoside (0.03 units). Reactions were incubated at 37oC for the indicated periods of time, and stopped by adding 4 µl of 98% formamide, 10 mm EDTA and 0.025% bromophenol blue. Samples were denaturiruet under 100oC for 5 minutes followed by rapid cooling on ice. Unreacted substance, as well as the reaction products were separated on 15% polyacrylamide/urea Sequeira gels and visualized using autoradiography. Oligonucleotide (-)-OddC at the 3'-end remained resistant to 3'-exonuclease, at least five times longer than other oligonucleotides.

Modifications and variations of the present invention in the treatment of malignant tumors will be obvious to experts in this field from the previous detailed description of the invention. Have in mind that such modifications and variations fall within the scope of the attached claims.

1. Pharmaceutical composition for treatment of a tumor in an animal host that contains an effective amount of-L-enantiomer of the formula

< / BR>
where R1and R2selected ENISA least 95% free of the corresponding D-enantiomer,

or its pharmaceutically acceptable salt in a pharmaceutically acceptable carrier.

2. The composition according to p. 1, where R, R1and R2represent hydrogen.

3. The composition according to p. 1, where the alkyl group is selected from the group consisting of methyl, ethyl, propyl, butyl, pentyl, hexyl, isopropyl, isobutyl, sec-butyl, tert-butyl and isopentyl.

4. The composition according to p. 1, where the acyl group represents-C(O)R, where R is a C1- C5alkyl group, phenyl or benzyl.

5. Composition under item 1 or 2, where the animal owner is the man.

6. The pharmaceutical composition according to paragraphs.1 - 5, where the carrier is acceptable for oral administration.

7. The pharmaceutical composition according to paragraphs.1 - 5, where the media presents in the form of capsules.

8. The pharmaceutical composition according to paragraphs.1 - 5, where the media presents in the form of tablets.

9. The pharmaceutical composition according to paragraphs.1 - 5, where the carrier is acceptable for intravenous administration.

10. The pharmaceutical composition according to paragraphs.1 - 5, where the carrier is acceptable for local or percutaneous injection.

11. The pharmaceutical composition according to paragraphs.1 - 5, where the introduction is parentalistically the composition according to p. 12, where the cancer is a prostate cancer.

14. The pharmaceutical composition according to p. 12, where the cancer is a leukemia.

15. The pharmaceutical composition according to p. 12, where the cancer is a colon cancer.

16. The pharmaceutical composition according to p. 12, where the cancer is a cancer of the bladder.

17. The pharmaceutical composition according to p. 12, where cancer is a liver cell cancer.

18. The pharmaceutical composition according to p. 12, where the cancer is a breast cancer.

19. The pharmaceutical composition according to p. 12, where the cancer is a lung cancer.

20. The pharmaceutical composition according to p. 12, where cancer is a nasopharyngeal cancer.

21. The pharmaceutical composition according to p. 12, where the cancer is a pancreatic cancer.

22. The pharmaceutical composition according to p. 12, where the cancer is an ovarian cancer.

23. The pharmaceutical composition according to p. 12, where the cancer is a lymphoma.

24. Pharmaceutical composition for treating cancer in an animal host that contains an effective amount of-L-enantiomer of the formula

< / BR>
where R1and R2selected from the group 95% free of the corresponding D-enantiomer,

or its pharmaceutically acceptable salt in a pharmaceutically acceptable carrier.

25. The pharmaceutical composition according to p. 24, where R is fluorine and R1and R2denote hydrogen.

26. The pharmaceutical composition under item 24 or 25, where the animal host is the man.

27. Pharmaceutical composition for p. 26, where the carrier is acceptable for oral administration.

28. Pharmaceutical composition for p. 26, where the media presents in the form of capsules.

29. Pharmaceutical composition for p. 26, where the media presents in the form of tablets.

30. Pharmaceutical composition for p. 26, where the introduction is parenteral.

31. Pharmaceutical composition for p. 26, where R denotes hydrogen.

32. The pharmaceutical composition according to p. 31 where the cancer is a prostate cancer.

33. The pharmaceutical composition according to p. 31 where the cancer is a leukemia.

34. The pharmaceutical composition according to p. 31 where the cancer is a colon cancer.

35. The pharmaceutical composition according to p. 31 where the cancer is a cancer of the bladder.

36. The pharmaceutical composition according to p. 31 where the cancer comes a breast cancer.

38. The pharmaceutical composition according to p. 31 where the cancer is a lung cancer.

39. The pharmaceutical composition according to p. 31 where the cancer is a nasopharyngeal cancer.

40. The pharmaceutical composition according to p. 31 where the cancer is a pancreatic cancer.

41. The pharmaceutical composition according to p. 31 where the cancer is an ovarian cancer.

42. The pharmaceutical composition according to p. 31 where the cancer is a lymphoma.

43. Pharmaceutical composition for treatment of a tumor in an animal host that contains an effective amount of the compounds of formula

< / BR>
where R1and R2selected from the group consisting of hydrogen, alkyl, acyl, monophosphate, diphosphate, and triphosphate,

or its pharmaceutically acceptable salt, optionally in a pharmaceutically acceptable carrier.

44. The pharmaceutical composition according to p. 43, where R1and R2denote hydrogen.

45. The method of obtaining 2-hydroxymethyl-5-(cytosine-1-yl)-1,3-dioxolane, including interaction of the compounds of formula

< / BR>
where R1ameans hydroxyamino group

with a reagent that converts oxoprop in 4th position brazilero ring aminopropoxy-4-yl)cytosine, characterized in that conduct protected interaction of 1,3-dioxolane of the formula

< / BR>
where R1a- benzoyl, R1B- acetyl,

protected, for example, siciliani, casinowin base with the use of a Lewis acid such as trimethylsilyltriflate, which is not ratemyserver target product.

47. (-)-(2S, 4S)-1-(2-Hydroxymethyl-1,3-dioxolane-4-yl)cytosine or its derivatives or salts, or (+)-enantiomer or racemic mixture and pharmaceutically acceptable derivatives and salts, as an active ingredient of a medicinal product for the treatment or prevention of tumors, including cancer, or to treat conditions with abnormal or undesirable cell proliferation.

48. (-)-(2S,4S)-1-(2-Hydroxymethyl-1,3-dioxolane-4-yl)cytosine, or its derivatives and salts, or (+)-enantiomer or racemic mixture and its pharmaceutically acceptable derivatives ingredient and salts as an active tool in drug therapy, for example, for the treatment or prevention of tumors, including cancer, or to treat conditions with abnormal or undesirable cell proliferation.

49. Pharmaceutical composition for the treatment of psoriasis containing hydrogen, alkyl, acyl, monophosphate, diphosphate, and triphosphate,

or its pharmaceutically acceptable salt, optionally in a pharmaceutically acceptable carrier.

Priority points:

06.09.94 on PP.1-46;

17.02.95 on PP.1-46;

05.09.95 on PP.47-49.

 

Same patents:

The invention relates to intermediate compounds used in the synthesis of CIS-nucleosides, their analogues and derivatives of nucleosides of high purity, the method of obtaining these intermediate compounds

The invention relates to new derivatives of benzofuranyl, possess valuable biological properties, in particular to derivatives of N-(3-benzofuranyl)urea, mixtures of their isomers, or individual isomers and their salts

The invention relates to benzofuran formula I

< / BR>
where R1denotes NH2, 1-piperazinil or 4-R3-piperazinil;

R2denotes H, Cl, Br, OH or OA;

R3denotes benzyl or itself known protective for the amine function group;

X denotes a CN, COON, COOA, COOPh, COOCH2Ph, COOPy, CONR4R5or CO-Het;

R4and R5each independently of one another denotes H, A or benzyl;

A denotes alkyl with 1-4 C-atoms;

Ph denotes phenyl;

Het represents imidazol-1-yl, triazole-1-yl or tetrazol-1-yl; and

Py denotes a pyridyl;

and their salts

The invention relates to organic chemistry, specifically to a method for producing 2-(furyl-2)-1,3-imidazolidine, which has astragalina punctigera and bactericidal activity

The invention relates to new substituted pyrimidinediamine or alkylating compounds, their pharmaceutically acceptable salts, hydrates, N-oxides and method for inhibition of reverse transcriptase of the virus

The invention relates to new biologically active compounds, methods of treating diseases with their use and pharmaceutical compositions based on these compounds

The invention relates to new sulfonamide of General formula I, where R1-R8A and B have the meanings indicated in the formula, which are inhibitors of endothelin and can be used for the treatment of diseases associated with the activity of endothelin, such as high blood pressure, as well as to pharmaceutical compositions based on

The invention relates to compounds intended for use in the pharmaceutical industry as active ingredients in the manufacture of medicines

FIELD: organic chemistry, medicine, pharmacy.

SUBSTANCE: invention relates to new derivatives of 5-phenylpyrimidine or their pharmaceutically acceptable acid-additive salts that elicit properties of antagonists of neuropeptide receptor neurokinin-1 (NK-1). This allows their applying for treatment of such diseases as Alzheimer's disease, cerebrospinal sclerosis, attenuating syndrome in morphine withdrawal, cardiovascular alterations and so on. Compounds of invention correspond to the general formula (I):

wherein R1 means hydrogen or halogen; R2 means hydrogen, halogen atom, (lower)-alkyl or (lower)-alkoxy-group; R3 means halogen atom, trifluoromethyl group, (lower)-alkoxy-group or (lower)-alkyl; R4/R4' mean independently hydrogen atom or (lower)-alkyl; R5 means (lower)-alkyl, (lower)-alkoxy-group, amino-group, hydroxyl group, hydroxy-(lower)-alkyl, -(CH2)n-piperazinyl substituted optionally with lower alkyl, -(CH)n-morpholinyl, -(CH2)n+1-imidazolyl, -O-(CH2)n+1-morpholinyl, -O-(CH2)n+1-piperidinyl, (lower)-alkylsulfanyl, (lower)-alkylsulfonyl, benzylamino-group, -NH-(CH2)n+1N(R4'')2, -(CH2)n-NH-(CH2)n+1N(R4'')2, -(CH2)n+1N(R4'')2 or -O-(CH2)n+1N(R4'')2 wherein R4'' means hydrogen atom or (lower)-alkyl; R6 means hydrogen atom; R2 and R6 or R1 and R6 in common with two ring carbon atoms can represent -CH=CH-CH=CH- under condition that n for R1 is 1; n means independently 0-2; X means -C(O)N(R4'')- or -N(R4'')C(O)-. Also, invention relates to a pharmaceutical composition.

EFFECT: valuable medicinal properties of compounds.

15 cl, 4 sch, 86 ex

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