Analogues of vitamin d3

 

(57) Abstract:

The described compounds of formula I:

where R1is hydrogen, R2- C1-C4alkyl, or R1- C1-C4alkyl, and R2is hydrogen; And CH2-CH2group; R3- C1-C4alkyl, hydroxy1-C4alkyl or fluorine WITH1-C4alkyl, and R4- C1-C4alkyl, hydroxy1-C4alkyl or fluorine WITH1-C4alkyl; compounds suitable for the treatment of breast cancer, prostate cancer, myeloid leukemia, benign tumors of the prostate, alopecia and osteoporosis. 3 C.p. f-crystals, 8 PL.

The invention relates to a compound of the formula

where

R1means hydrogen or alkyl group

R2means hydrogen or alkyl group, or

R1, R2and C20together represent cyclopropyl;

And means

R3means alkyl, hydroxyalkyl or foralkyl and

R4means alkyl, hydroxyalkyl or foralkyl.

It was found that the compounds of formula I cause inhibition of proliferation in cell lines tumors of the prostate gland is part of the gland, breast cancer and for the treatment of leukemia.

It was also found that the compounds of formula I possess antiandrogenna activity. Therefore, the compounds of formula I are suitable for the treatment of benign tumors of the prostate, alopecia and prostate cancer.

It was also found that the compounds of formula I have activity, which make them suitable for the treatment of diseases of the sebaceous glands, such as acne or seborrheic dermatitis.

It was also found that the compounds of formula I have activity makes these compounds suitable for the treatment of osteoporosis.

Detailed description of the invention

Used herein, the term "alkyl group" means an alkyl group with straight or branched chain containing 1-4 carbon atoms, for example methyl, ethyl, propyl, isopropyl, butyl, tert-butyl and the like groups. The term "hydroxyalkyl" means an alkyl group with a substituent in the form of hydroxyl at any of the carbon atoms of the alkyl group. The term "foralkyl" means an alkyl group containing one, two or three fluorine atom as substituents at any of the carbon atoms of the alkyl group.

In these formulas, R is shaped solid line (), pointing to the Deputy located above the plane of the molecule, and a wedge-shaped dotted line () indicating a Deputy under the plane of the molecule.

Preferably R1means hydrogen and R2means alkyl or R1means alkyl and R2means hydrogen. More preferably R1means hydrogen and R2means methyl, or R1means methyl and R2means hydrogen.

Preferably R3and R4means independently alkyl, hydroxyalkyl or triptorelin. More preferably R3and R4mean independently from each other methyl, hydroxymethyl or trifluoromethyl.

Preferably And means

The most preferred compound of formula I is 1,25-dihydroxy-16-ene-5,6-TRANS-cholecalciferol.

The compounds of formula I receive, as described below, in full accordance with the below chemical scheme.

The chemical scheme

R5means hydrogen or trimethylsilyl.

In the above chemical scheme, where Ph means phenyl, the compound of formula II, which is [3S-(1E, 3, 5)]-[2-[3,5-[[bis(1,1-(dimethylethyl)dimethylsilane]oxy]-2-methyltrichlorosilane]aticio carried out at a temperature from -60°C to -90°C in a polar aprotic organic solvent, as, for example, simple anhydrous ether or more preferably anhydrous tetrahydrofuran, in the presence of a strong base, such as, for example, litearcy, such as utility.

The protective group of compounds of formula IA can be removed by reaction with fluoride salts, such as tetrabutylammonium fluoride in a polar organic solvent, for example in a simple ether or more preferably tetrahydrofuran, and receive a corresponding compound of formula I.

The compound of formula II get as hereafter described in examples 1-6 or, alternatively, as described in examples 7 and 8.

The compounds of formula III are known (for example, from U.S. patent No. 5087619) or they can be obtained in accordance with known methods.

The useful activity of compounds of the formula I as tools for the treatment of prostate cancer, breast cancer and for the treatment of leukemia can be demonstrated by the following research methods, well known in this field.

The materials selected for use here and the techniques applied were as follows:

Cell line. Line of breast cancer cells (MCF-7), line of prostate cancer cells (LNCaP) and cell line m is the red Needle (DMEM) with 10% amniotic calf serum (FCS); LNCaP and HL-60 were cultured in medium RPMI 1640 with 10% FCS. All three cell lines were maintained in an incubator with 37°C containing 5% carbon dioxide.

Derivatives of vitamin D3. Derivatives of vitamin D3was dissolved in absolute ethanol at a concentration of 10-3M, such solutions were used as initial solutions, which were stored at -20°C and protected from light. For in vitro use of the compounds were diluted in DMEM or RPMI medium. For use in vivo, the compounds were diluted in phosphate buffered saline (SFR). An aliquot was used only once and LNCaP cells were subjected to the action of trypsin. Washed unicellular suspension of cells was counted and placed in 24-cell tablet with a flat bottom with a total of 110-3cells/cell in a total volume of 400 μl/cell. Feeding layer was obtained with the agar, which was balanced at 42°C. Before this stage in connection with a pipette and placed in a cell. After incubation were counted colonies. All experiments were performed at least three times using triple tablets for each experimental point.

The levels of serum calcium in vivo. Twenty-eight male Balb/c mice aged 8 to 9 weeks coderay were intraperitoneally injected with every day (except Saturday and Sunday) or a derivative of vitamin D3or diluent (100 μl/mouse) for 3 weeks. Doses of 1,25-dihydroxy-16-ene-5,6-TRANS-D3consisted of 0.1, 0.5, 1.0, and 2.0 µg. Doses of 1,25-dihydroxy-D3was 0.1 μg/mouse. Control mice were injected with 100 μl SFR. Calcium in serum was measured weekly using a quantitative colorimetric determination.

Experiments on pulse. Cells MCF-7 were incubated in liquid culture with 10-71,25-dihydroxy-D3or 1,25-dihydroxy-16-ene-5,6-TRANS-D3with different duration. After incubation, these cells were carefully washed twice SFR and viable cells were counted and placed in 24-cell tablet for analysis of colonies in soft agar as described above.

The cell cycle analysis using flow cytometry. The cell cycle analysis was performed on MCF-7 cells, incubated for 4 days with either 1,25-dihydroxy-D3or 1,25-dihydroxy-16-ene-5,6-TRANS-D3at a concentration of 10-7M. the Cells were fixed in chilled methanol overnight before staining with 50 μg/ml of propyliodide, 1 mg/ml of ribonuclease (100 units/ml) and 0.1% NP40. Analysis was performed immediately after staining. All experiments Provo is riteria the student.

Analysis of Western blot testing. Cells were twice washed SFR, suspended in buffer for lysis (50 mm Tris, pH 8.0, 150 mm such as NaCl, 0.1% sodium dodecyl sulfate, 0.5% desoxycholate sodium, 1% NP40, 100 μg/ml fluoride phenylmethylsulfonyl, 2 μg/ml Aprotinin, 1 μg/ml of pepstatin and 10 μg/ml leupeptin) and placed on ice for 30 minutes. After centrifugation at 15000 g for 20 minutes at 4°C and collected the supernatant. Quantitatively estimated protein concentration. All lysates (15 μg) were separated using polyacrylamide gel with 15% sodium dodecyl sulfate, carried on a fixed membrane from polyvinylidenedifluoride and subjected to analysis using anti-p27kiplrabbit polyclonal antibodies and mouse monoclonal antibodies against actin. The blots showed.

Telomerase activity. To determine the relative telomerase activity of the tested protocols amplification of the telomeric sequence (TRAP). For the human telomerase reverse transcriptase (hTERT) has allocated all of the RNA from HL-cells that were treated or 1,25-dihydroxy-D3or 1,25-dihydroxy-16-ene-5,6-TRANS-D3(10-9, 10-8and 10-7M) for 4 days using a single-phase solution of phenol and isotis the random review of the basic structures. cDNA was amplified using seed-specific hTERT gene or gene GAPDH, was used as control. The seed used for hTERT, were: 5’-CGGAAGAGTGTCTGGAGCAA-3’ (sense) /sequence identified No. 1, (Ate. ID. No. 1)/ 5’-GGATGAAGCGGAGTCTGGA-3’ (antisense) (Th. ID. No. 2). Thermal cycles were performed at 94°C for 90 seconds, then 33 cycles 95°C for 20 seconds, 68 PP. for 40 seconds and 72°C for 30 seconds. The seed for gene GAPDH were 5’-GGGGGGGGG-3’ (sense) (Th. ID. No. 3) and 5’-CAAAGTTGTCATGGATGACC-3’ (antisense) (Th. ID. No. 4). Conditions of GAPDH amplification were: 94°C for 2 min, 26 cycles with a temperature of 94°C for 30 seconds, 62°C for 40 seconds, 72°C for 60 seconds, then 72°C for 4 minutes. The products of polymerase chain reaction were subjected to electrophoresis on 1% agarose gel and stained with ethidiumbromid.

The effect of analogues of vitamin D3on analysis of clonogenicity. Cells LNCaP, MCF-7 and HL-60 were cloned in soft agar in the presence of analogues of vitamin 3at a concentration of from 10-11up to 10-7M Was obtained in a dose-dependent curves and was found to be the effective dose (ED50) that caused 50% inhibition of the radio three cell lines depending on the dose. ED501,25-dihydroxy-16-ene-5,6-TRANS-D3was 1,410-9M for LNCaP cells, 4,310-9M for MCF-7 cells and 3,010-11M for cells HL-60, this compound was approximately 10-100 times more active than 1,25-(dihydroxy)-D3.

The levels of serum calcium in vivo. Because hypercalcemia is the main form of toxicity associated with derivatives of vitamin D3, calciumcalcium effects of 1,25-dihydroxy-D3compared with those for 1,25-dihydroxy-16-ene-5,6-TRANS-D3. All mice survived the study within three weeks. All mice treated with 0.1 μg 1,25-dihydroxy-D3, hypercalcemia was observed with levels of calcium in the serum of approximately 12 mg/DL (norm 8.5-10.5 mg/DL). Conversely, in mice treated with 1,25-dihydroxy-16-ene-5,6-TRANS-D3(0.1 to 2.0 µg/mouse) was found almost the same level of calcium (8-10 mg/DL) as that of the control mice (8-9 mg/DL).

Experiments on pulse. To study whether the count of clonogenic inhibition of proliferation analogues of vitamin D3reversible, experiments were carried out on the pulse. Cells MCF-7 were subjected to either 1,25-dihydroxy-16-ene-5,6-TRANS-D3or 1,25-dihydroxy-D3with various Pro is steverivonia. About forty and thirty percent count of clonogenic cells inhibited on the 4th day of exposure to 1,25-dihydroxy-16-ene-5,6-TRANS-D3and 1,25-dihydroxy-D3respectively.

Analysis of the cell cycle. Determined the effect of 1,25-dihydroxy-16-ene-5,6-TRANS-D3and 1,25-dihydroxy-D3(10-74 days) on the cell cycle of MCF-7 cells. A significant increase (P0,05) the number of cells in the phase of G0-G1occurred with a concomitant decrease in the relative amount of cells in phase s

Analysis of Western blot testing. Cyclin-dependent kinase inhibitors, known as P21wafland p27kiplable to inhibit the activity cilincing and, thus, slow down the development of cells within the cell cycle. Control cells MCF-7, as it had a medium level of expression of p21wafland p27kiplas determined by analysis of Western blot testing. Impact in one day 1,25-dihydroxy-16-ene-5,6-TRANS-D3(10-7M) increased the expression of P21wafland p27kiplapproximately 3.2-3.5 times, while in the cultures with 1,25-dihydroxy-D3(10-7M) observed the increased expression of p21wafland p27kiplabout 1.6-1.8 times. The effect on the cells MCF-7 1,25-dihydroxy-16-ene-5,6-is and p27kiplrespectively. 1,25-Dihydroxy-D3(10-7M, 3 days) increased the expression of p21wafland p27kipl4.8 times and 3.3 times, respectively.

Investigated the dose-dependent effect of derivatives of vitamin D3on the expression of p27kiplcells HL-60. And 1,25-dihydroxy-D3and 1,25-dihydroxy-16-ene-5,6-TRANS-D3increased regulation of the expression of p27kipland these levels were significantly increased after incubation with 1,25-dihydroxy-16-ene-5,6-TRANS-D3(4 days, 10-9M). The levels of p27kiplwas significantly increased when cells HL-60 were cultured with 10-8-10-7M 1,25-dihydroxy-D3.

Telomerase activity. Applying the analysis TRAP, estimated the effect of 1,25-dihydroxy-16-ene-5,6-TRANS-D3and 1,25-dihydroxy-D3(10-9-10-7M, 4 days) on telomerase activity. Telomerase activity was significantly decreased in cells HL-60, cultivated with either 1,25-dihydroxy-16-ene-5,6-TRANS-D3, 10-9M, or 1,25-dihydroxy-D3, 10-7M

The effect of analogues of vitamin D3on the expression of hTERT in cells HL-60 was evaluated using RT-PCR. And 1,25-dihydroxy-16-ene-5,6-TRANS-D3and 1,25-dihydroxy-D3inhibited the expression of hTERT mRNA, depending on the dose with almost full ing is hydroxy-D3.

The useful activity of compounds of the formula I as tools for the treatment of benign prostate tumors can be demonstrated in the following ways research, known in this field.

In castrated stimulated by testosterone, the male Syrian hamsters were evaluated antiandrogenna activity of 1,25-dihydroxy-16-ene-5,6-TRANS-D3. Studies have shown that 1,25-dihydroxy-16-ene-5,6-TRANS-D3 significantly suppressed in these animals induced by androgen hypertrophy of seminal vesicles and ventral prostate, whereas 1,25-dihydroxy-D3was inactive in non-toxic doses (1 μg).

Castrated male Syrian hamsters were daily injected subcutaneously with 20 μg of testosterone propionate and 1 μg 1,25-dihydroxy-16-ene-5,6-TRANS-D3for 14 consecutive days. Both compounds were injected in each case in the media of 0.2 ml of sesame oil. An autopsy was performed on the 15th day. The prostate gland and seminal vesicles were removed, drained and weighed. Analyzed the data for statistical significance using student's criterion, and the results were expressed as percentage inhibition of stimulated response.

Ingoio hamsters are presented in table.1

Materials and methods

Animals and treatment:

Used the three-month adult female rats. After a week of adaptation, the animals were grouped according to body weight and were treated by oral administration of 1 ml/kg/day of 1,25-dihydroxy-16-ene-5,6-TRANS-D3in several concentrations. On the seventh day of dosing, animals were bled and determined the level of calcium in the serum.

Preparation of compounds

The compound was dissolved in 200 ml of ethanol installed the fortress to obtain a concentration of 100 ág/ml Media in the form of sesame oil and a solution of the compound in ethanol was prepared to the highest dose concentration, then was evaporated in a rotary evaporator at 37°C to remove the ethanol. The amount of dose was calculated by considering the average body weight in the group, which is the dosing. Serial cultivation dissolved in the media connections are made to the appropriate dose concentrations.

Collection of serum and determination

Blood (1.5 ml) was collected from each animal by puncture of the eye with anesthetic ether on the seventh day of dosing. Blood was collected in separator tubes for serum, centrifuged at 2000 rpm for 1 is colorimetrically analysis.

Results

The levels of serum calcium are shown in table.2.

The levels of serum calcium were within normal range for groups treated with the dose of 2 mcg/kg/day in the treatment of 1,25-dihydroxy-16-ene-5,6-TRANS-D3.

Materials and methods

Animals and treatment:

Three-month adult female rats were subjected to bilateral oophorectomy or simulated surgical intervention. After a week of adaptation, the animals were grouped according to body weight and were treated by oral administration of 1 mg/kg concentration.

The amount of dose was calculated on a weekly basis, considering the average body weight in the group. Dosing was started 17 days after surgery and continued for 19 days. On the 20th day animals were exposed painless death by inhalation of carbon dioxide and dissected the left thigh.

Total calcium thighs:

When the autopsy was dissected left thigh animals from all groups and was removed with a soft cloth. The bones were measured and cut in half in the middle of the diaphysis; then the distal section cut in half lengthwise after removal of the epiphysis. The bone marrow was washed and calcitrate analyzed, using quantitative colorimetric determination of calcium. The results were expressed as the average total calcium in the bones in mg/distal half of the femur (DHF)±serum.

Collection of serum and determination

Blood (1.5 ml) was collected from each animal at the puncture of the eye under ether anesthesia on day 7 and day 18 of dosing. Blood was collected in separator tubes for serum, centrifuged at 2000 rpm for 15 minutes and then was selected aliquot of serum samples for determinations of calcium. Calcium in serum were determined using colorimetric analysis.

Usage:

To confirm the effect of oophorectomy on the content of calcium in the bones were compared using student's criterion groups subjected to simulated surgery /sham/ and undergone oophorectomy /ovx/ processing filler. Group after oophorectomy were compared using one-way analysis of variance (ANOVA) with subsequent processing by the method of least squares Fisher for comparing each treated group with the treated filler of the group when the overall effect was statistically significant.

Results

Impact on cal is a useful activity of compounds of the formula I as tools for the treatment of diseases of the sebaceous glands, as demonstrated by the following testing methods and techniques known in this field.

Was dissolved with 200 μl of 1,25-dihydroxy-16-ene-5,6-TRANS-D3in propylene glycol, was daily injected (5 days per week) with food male Golden Syrian hamsters. Animals were killed after 4 weeks and ears were subjected to processing for histological evaluation. The area of the sebaceous glands were histologically measured on the obtained cross sections of the ear using image analysis.

The results are shown in table.5.

Also was investigated total lipid fraction from one of the ears (extraction with an organic solvent, followed by weighing the remaining lipid material).

The results are shown in table.6.

The compounds of formula I can be administered orally for the treatment of breast cancer, prostate cancer or leukemia people who need such treatment. More specifically, the compounds of formula I can be introduced to such treatment oral adult in doses ranging from about 1 to 20 μg per day.

The compounds of formula I can be administered orally for the treatment of benign tumors of the prostate and alopecia people who flax adult doses, within about 1 to 20 μg per day.

The compounds of formula I can be administered orally for the treatment of osteoporosis in humans at a dose of about 1 to 20 μg per day.

The compounds of formula I can be applied topically for the treatment of alopecia in humans in need of such treatment. More specifically, the compounds of formula I can be applied to such treatment topically in dosages that are in the range from about 5 to 50 μg per gram dosage forms for topical use on the day.

The compounds of formula I can be administered orally for the treatment of diseases of the sebaceous glands in humans at a dose of about 1 to 20 μg per day.

Oral dosage forms comprising compounds of the formula I according to the invention can be incorporated into capsules, tablets and similar forms together with a pharmaceutically acceptable substances-carriers.

Examples of pharmaceutically acceptable carriers that may be incorporated into capsules and similar forms are the following: a binder, as, for example, tragakant, acacia, corn starch or gelatin; an excipient, such as, for example, secondary acidic calcium phosphate; crumbling tool, such as corn KRA is tion; sweetener, such as sucrose, lactose or saccharin; corrigent, such as peppermint, oil of Grushenka or cherry. Various other substances may be present as a shell or to otherwise modify the physical form of the standard dose. For example, tablets may be coated with shellac, sugar or both. A syrup or elixir may contain the active compound, sucrose as a sweetener, methyl and propyl esters of parenting acid as preservatives, a dye and flavoring, giving, for example, the taste of cherry or orange.

Dosage forms for topical application, containing the compounds of formula I according to the invention include ointments and creams containing a finished dosage form that has oily, absorbable, water-soluble basics and fundamentals of emulsion type, such as petrolatum, lanolin, polyethylene glycols and the like compounds.

Lotions are liquid preparations and differ from simple solutions by the presence of water or giprospirtvino preparations containing finely ground substances. Lotions can contain suspendresume or dispersing means, such as derivatives of cellulose, cadient in the filler, composed of water, alcohol, glycerol and the like substances.

Gels are semi-solid preparations prepared by thickening the solution or suspension of the active ingredient in the raw filler. The fillers, which may be aqueous or anhydrous, get thick when using a freezing gel means, as, for example, carboxypolymethylene, and neutralized to a proper gel consistency when using alkali, such as sodium hydroxide, and amines, for example polyethyleneamine.

Used here, the term "local" refers to the use of the active ingredient incorporated in a suitable pharmaceutical carrier, and applied at the site of inflammation for the provision of local action. Accordingly, compositions for topical application include those pharmaceutical forms in which the substance is used externally by direct contact with the skin. Dosage forms for topical application include gels, creams, lotions, ointments, powders, aerosols and other common forms of drug application on the skin, obtained by mixing compounds of the formula I with known pharmaceutical carriers for local use.Example 1

(2R,3S,5S,7S)-2-[5.7-Bis[(1,1-dimethylethyl)dimethylsilane]oxy]-4-methylene-1-oxaspiro[2.5]octane-2-acetate

To stir magnetic stirrer, a solution of 18.5 g (0,0523 mol) (2R, 3S, 5S, 7S)-5-hydroxy-4-methylene-7-[(1,1-dimethylethyl)dimethylsiloxy]-1-oxaspiro-[2,5]octane-2-acetate (Y. Kiegiel, P. M. Wovkulich and M. R. Uskokovic, Tetrahedron Letters, 32. pgs.6057-6060 (1991)) and 6.8 g (0,099 mole) of imidazole in 50 ml of dimethylformamide was added in an argon atmosphere of 9.8 g (0,065 mol) tert-butyldimethylsilyloxy.

The reaction mixture was stirred for 5 hours, stopped the reaction by adding 5 ml of water, stirred for 30 minutes and poured into 400 ml of water. Then the mixture was extracted with hexane (2500 ml) and ethyl ether (2500 ml). The organic layers were combined, washed with 300 ml of water, dried over sodium sulfate and evaporated. After chromatography on a column of silica gel received 22,31 g (92%) specified in the title compounds as colorless oils.

Example 2

[3S-(1E,3,5)]-2-[3,5-Bis[[(1,1-dimethylethyl)dimethylsilane]oxy]-2-methylene-cyclohexylidene]acetate

In a three-liter three-neck flask equipped with inlet for argon, mechanical stirrer and thermometer, was loaded with 500 ml of tetrahydrofuran and cooled to -60°C in the bath with dry Lido is the temperature -60°C, and then carried out a rapid dropwise addition of 200 ml of 1.6 M n-butyl lithium in hexane, maintaining the temperature below -45°C (about 5 minutes). Bath with dry ice in acetone was replaced by a bath of water and ice, allowing to rise the temperature to 5°C. the Color changed from blue to khaki until black color with a reddish tint. After 30 minutes was added at 5°C rapidly dropwise within 3 minutes the solution 22,31 g (0,04884 mol) (2R, 3S, 5S, 7S)-2-[5,7-bis[(1,1-dimethylethyl)-dimethylsilane]oxy]-4-methylene-1-oxaspiro[2,5]octane-2-acetate in 50 ml of hexane. After 4 hours the reaction mixture was diluted with 2 liters of hexane, filtered through a layer of silica gel, which was washed 3500 ml of hexane-ethyl acetate 9:1, and evaporated. The residue was chromatographically on the column with 75 g of silica gel, received 22,56 g technical product. The medium pressure chromatography on a column of silica gel and elution with a mixture of 100:1 dichloromethane-ethyl acetate led to 17,42 g (80,1%) specified in the title compound and a small amount of relevant 1Z-epimer.

Example 3

[3S-(1E,3,5)]-2-[3,5-Bis[[(1,1-dimethylethyl)dimethylsilane]oxy]-2-methyltrichlorosilane]acetate (E-diene)

[3S-(1Z,3,5)]-2-[3,5-bis[[(1,1-dimethylethyl)dimethylsilane]oxy]-2-methylenecycloartanol>6(blue solution) was then added 337,5 ml 1.43 M n-utility in hexane (with such speed that the temperature in the reaction mixture did not rise above -20°C). Then the mixture was allowed to warm to room temperature. Was added dropwise a solution of 24.5 g (53.6 mmol) of (2S, 3R, 5S, 7S)-2-[5,7-bis[(1,1-dimethylethyl)-dimethylsilane]oxy]-4-methylene-1-oxaspiro[2,5]octane-2-acetate in 65 ml of THF and the mixture was stirred for 4 hours. The mixture was diluted with 600 ml of pentane and filtered through a layer of silica gel in 4 cm, rinsing with hexane/ethyl acetate (19:1), was obtained after evaporation of the volatiles under reduced pressure of 27 g of the crude mixture of dienes. Further purification using chromatography with elution with hexane/ethyl acetate (25:1) resulted in a 21.6 g (91%) of a mixture (2:3) Z/E-dienes (mentioned in the title compound), which was separated by chromatography on silica gel (hexane/ethyl acetate 40:1).

Z-diene,1H-NMR : 0,052 (s, 6H), is 0.06 (s, 6H), of 0.87 (s, 9H), to 0.89 (s, 9H), 1,74-1,90 (m, 2H), 2,04 (s, 3H), of 2.20 (dd, J=6,0, to 12.8 Hz, 1H), 2,41 (d, J=11,1 Hz, 1H), 4,19 (m, 1H), 4,42 (m, 1H), br4.61 (dd, J=7,3, to 12.1 Hz, 1H), and 4.68 (dd, J=7,3, to 12.1 Hz, 1H), 4,80 (s, 1H), 5,20 (s, 1H), vs. 5.47 (t, J=7,2 Hz, 1H). []25D=+1,2° (c=0.4, EtOH). Analysis calculated for C23H44O4Si2: 62,67; N 10,06; found: 62,55; N 10,33.

E-diene,1H-NMR : 0,046 (s, 3H), 0,057 (s, 3H), 0,065 (s, 6H), 0.88 to, 2,9 Hz, 1H) and 4.65 (dd, J=7,2, and 12.9 Hz, 1H), 4.95 points (s, 1H), 4,99 (s, 1H), 5,69 (t, J=7,2 Hz, 1H). []25D=+8,0° (C=0,5, tO). Analysis calculated for C23H44O4Si2: 62,67; N 10,06; found: 62,42; N 10,01.

Example 4

[3S-(1E,3,5)]-2-[3,5-Bis[[(1,1-dimethylethyl)dimethylsilane]oxy]-2-methylene-cyclohexylidene]ethanol

To stir magnetic stirrer, the solution 10,84 g (0,0246 mole) of [3S-(1E,3,5)]-2-[3,5-bis[[(1,1-dimethylethyl)dimethylsilane]oxy]-2-methyltrichlorosilane]-acetic acid ethyl ester in 100 ml of methanol was added in an argon atmosphere 3.5 g of pellets of sodium hydroxide and the reaction mixture was stirred under argon for 3 hours. Then the mixture was evaporated in vacuum to a volume of 50 ml, diluted with 500 ml of water, was extracted with 2500 ml of hexane-diethyl ether 1:1. The organic layer was washed with water, dried over sodium sulfate and evaporated. Received 9.80 g (100%) specified in the title compound in the form of a solid white color.

Example 5

1R-(1,3,5 E)-[[5-(2-Chloraniline)-4-methylene-1,3-cyclohexanediyl]bis(oxy)]bis(1,1-dimethylethyl)dimethylsilane

To stir the solution to 6.67 g (0,050 mole) N-chlorosuccinimide in 150 ml of dichloromethane, cooled to 2°C in the bath with ice in acetone, in an atmosphere of argon was added dropwise over 2 minutes 4 the Dom in acetone and established the temperature of the reaction mixture to -20°C. Was added a solution of 9.8 g (0,0246 mole) of [3S-(1E,3,5)]-2-[3,5-bis[[(1,1-dimethylethyl)-dimethylsilane]oxy]-2-methyltrichlorosilane]ethanol in 60 ml of dichloromethane. After 15 minutes the cooling bath was removed and the reaction mixture was stirred for 50 minutes and then transferred into a separating funnel containing 500 ml of water. The mixture was extracted 2350 ml of hexane. The organic layer was washed with 500 ml of water, dried over sodium sulfate and evaporated, got 10,49 g of product as a yellow liquid. After purification using flash chromatography got cleaned specified in the title substance as a colorless oil.

NMR (CDCl3) : 0,03 (s, 3H), of 0.05 (s, 3H), of 0.07 (s, 6H), of 0.87 (s, 9H), to 0.89 (s, 9H), 1.70 to was 1.94 (m, 2H), 2,34 (m, 2H), 4,13 (m, 2H), 4,28 (m, 1H), 4.53-in (m, 1H), 5,00 (m, 1H), to 5.03 (m, 1H), 5,78 (tm, J=8 Hz, 1H).

Example 6

[3S-(1E,3,5)]-[2-[3,5-Bis[[(1,1-dimethylethyl)dimethylsilane]oxy]-2-methylene-cyclohexylidene]ethyl]diphenylphosphine

In a three-neck flask with a volume of 1 l, equipped with inlet for argon, thermometer and mechanical stirrer, was loaded solution 10,26 g (0,0246 mole) of 1R-(1,3,5 E)-[[5-(2-chloraniline)-4-methylene-1,3-cyclohexanediyl]bis(oxy)]bis-(1,1-dimethylethyl)dimethylsilane in 100 ml of fresh anhydrous tetrahydrofuran and cooled in a bath of dry ice and Aceto color required 60 ml of reagent. After stirring for 1 hour at-65S were added 10 ml of water and removed the bath for cooling. The reaction mixture was bleached. Then there was added 200 ml of dichloromethane and then quickly 200 ml of an aqueous solution containing 10 ml of 30% hydrogen peroxide. After 1 hour was added 13.5 g of sodium sulfite, 100 ml of brine and 200 ml of dichloromethane. After thorough shaking, the phases were separated and the aqueous phase is washed with 200 ml of dichloromethane. The organic phase was washed with 200 ml brine. The combined organic layers were dried over sodium sulfate, filtered and evaporated, got 16,33 g technical product. This technical product was purified using medium pressure chromatography (silica gel G-60), received 12,54 g (87%) specified in the title compound as white crystals.

NMR (CDCl3) : 0,02 (s, 3H), of 0.07 (s, 6H), and 0.08 (s, 3H), 0,84 (s, 18H), to 1.76 (m, 2H), 2,98 is 3.15 (m, 2H), 4,06 (m, 1H), 4,37 (m, 1H), 4.53-in (m, 1H), 4.72 in (m, 1H), 4,79 (m, 1H), 7,45 (m, 6H), 7,72 (m, 4H).

Example 7

Methyl ether E-(3S,5R)-[3,5-bis(tert-butyldimethylsilyloxy)-2-methyltrichlorosilane]acetic acid (Ro 65-8821)

A solution of 4.45 g (0,01043 mole) of the methyl ester of Z-(3S, 5R)-[3,5-bis(tertbutyl-dimethylsiloxy)-2-methyltrichlorosilane]acetic acid (X) (A. Mowrino et al., Tetrahedron Letters, 38, pgs. 4%) specified in the title compounds as colorless oils.

NMR (CDCl3) : 0,06 (s, 3H), of 0.07 (s, 9H), of 0.85 (s, 9H), to 0.89 (s, 9H), to 1.76 (m, 1H), of 1.84 (m, 1H), 2,70 (m, 1H), 3,36 (m, 1H), 3,70 (s, 3H), 4.26 deaths (m, 1H), 4,58 (m, 1H), 5,07 (m, 2H), 5,91 (ush. s, 1H).

Example 8

[3S-(1E,3,5)]-2-[3,5-Bis[[(1,1-dimethylethyl)dimethylsilane]oxy]-2-methylene-cyclohexylidene]ethanol

To a solution of 4.25 g (0,00996 mole) methyl ester E-(3S,5R)-[3,5-bis(tert-butyl-dimethylsiloxy)-2-methyltrichlorosilane]acetic acid in 100 ml of toluene at -78°C was added dropwise 25 ml of 1.25 M diisobutylaluminium (0,03 mole) and the reaction mixture was stirred for 1 hour. After addition of 5 ml of methanol, the reaction mixture was allowed to warm to room temperature. The mixture is then diluted with 150 ml of 2 M aqueous solution of potassium-sodium salt of tartaric acid and intensively stirred. The organic phase was separated, dried over sodium sulfate and evaporated to dryness. Technical product was purified using flash chromatography using hexane-ethyl acetate 8:2, was obtained 2.8 g (70%) specified in the title compounds as a colorless waxy solid.

NMR (CDCl3) : of 0.05 (s, 3H), of 0.07 (s, N), to 0.88 (s, 9H), of 0.91 (s, N), of 1.74 (m, 1H), 2.06 to (m, 1H), 2.26 and (dm, J=13,6 Hz, 1H), 2.40 a (dd, J=13,6, and 5.2 Hz, 1H), 4,30-40,11 (m, 3H), 4.53-in (m, 1H), to 4.98 (m, 1H), 5,00 (m, 1H), 5,80 (tm, J=7 Hz, 1H).

Example 9

1,25-Dihydroxy-16-ene-5,6-tsili]oxy]-2-methyltrichlorosilane]ethyl]diphenyl-phosphine oxide in 10 ml of anhydrous tetrahydrofuran was treated at -78°C by adding dropwise under argon 0,83 ml (0,00133 mol) of 1.6 M n-utility in hexane. To the thus obtained red solution was added dropwise under argon for 10 minutes a solution of 280 mg (0,000803 mole) of [3aR-[1(R),3A,7a]]-1-[1,5-dimethyl-5-[(trimethylsilyl)oxy]hexyl]-3,3 and,5,6,7,7 and hexahydro-7a-methyl-4H-inden-4-it is in 5 ml of tetrahydrofuran. The reaction mixture was stirred at -78°C for 90 minutes, then the reaction was stopped by adding 40 ml of a mixture (1:1) 2 N. potassium tartrate of sodium and 2 n potassium bicarbonate and gave the reaction mixture to warm to room temperature. The reaction mixture was extracted (3100 ml) ethyl acetate. The organic layers were washed three times with water/salt solution, dried over sodium sulfate and evaporated to dryness. This technical product was purified using flash chromatography on a column of 40 mm 6’ silica gel with elution with hexane-ethyl acetate 40:1, received 278 mg traileraverage derivative specified in the title compound and 140 mg of the original ketone.

A solution of 278 mg (0,000389 mole) of this traileraverage intermediate product in 8 ml of anhydrous tetrahydrofuran was treated with 1.9 ml (1.9 mmole) of 1 M solution of tetrabutylammonium in tetrahydrofuran under argon for 17 hours. The reaction was stopped by adding 6 ml of water and paramasivam. The organic layers were washed four times with water/salt solution, dried over sodium sulfate and evaporated to dryness. Technical product, 192 mg, was purified using flash chromatography on a column of silica gel, which was pre-washed with 1% solution of triethylamine in ethyl acetate (300 ml); elution was performed with ethyl acetate. When this was received 152 mg crystalline compounds specified in the title. A sample, recrystallized from tetrahydrofuran: methylformate (0,3:7), had tPL95-100°C. []25D+160,5° (EtOH, c=0,20).max272/3 nm ( 20600).

Example 10

Soft gelatin capsule

Part I is presented in table.7.

Method of preparation:

1. Preparing a suspension of OSH, BGA in miglioli 812. Heated to approximately 50°C and stirred until dissolution.

2. Dissolve 1,25-dihydroxy-16-ene-5,6-TRANS-D3in solution from operations 1 at 50°C.

3. Cool the solution from operations 2 to room temperature.

4. Fill soft gelatin capsules solution from operations 3.

Note: All operations of the cooking is performed in the nitrogen atmosphere, protecting from light.

Example 11

Soft gelatin capsule

Part II of t the e 812. Heated to approximately 50°C and stirred until dissolution.

2. Dissolve 1,25-dihydroxy-16-ene-5,6-TRANS-D3in solution from operations 1 at 50°C.

3. Cool the solution from operations 2 to room temperature.

4. Fill soft gelatin capsules solution from operations 3.

Note: All operations of the cooking is performed in the nitrogen atmosphere, protecting from light.

1. The compound of the formula I

where R1means hydrogen, and R2means1-C4alkyl; or

R1means1-C4alkyl, and R2means hydrogen;

And means-CH2-CH2group

R3means1-C4alkyl, hydroxy1-C4alkyl or fluorine WITH1-C4alkyl; and

R4means1-C4alkyl, hydroxy1-C4alkyl or fluorine WITH1-C4alkyl.

2. Connection on p. 1, where r1means hydrogen and R2means methyl, or R1means methyl and R2means hydrogen.

3. Connection on p. 2, where R3and R4mean independently from each other methyl, hydroxymethyl or trifluoromethyl.

4. Connection on p. 3, pre

 

Same patents:

The invention relates to a derivative of vitamin D formula (1):

Formula 1

where X = -O - or-S- ; m = 1, 2, 3; R1and R2-H or alkyl; R4-H; R5Is H, OH or R4and R5together form a double 16, 17 connection, R3- YR8where Y is-O - or-S-,

R8-H, alkyl, possibly substituted by F or cycloalkyl, or-NR9R10where R9and R10-H, alkyl, possibly substituted by F or cycloalkyl, R6-OH, possibly substituted, R7-H or a protective group

The invention relates to new derivatives of vitamin D General formula I

< / BR>
where Y1- OH, C1-12alkanoyloxy or optionally substituted benzoyloxy, Y2- H, C1-12alkanoyl or optionally substituted benzoline group, R1and R2together ekzoticheskaya methylene group, R3and R4independently - H, C1-4alkyl, Q - C1-3alkylen, possibly substituted inoris HE in the group, which, in turn, can be etherification, R5and R6at the same time C1-4alkyl, or R5and R6together with carbon atom C-25 form cyclopropyl group, Z is a 5-6-membered aromatic Carbo - or heterocycle, such as phenyl, oxazole, thiazole, furan, thiophene, pyrrole, isoxazol, pyrazole, triazole, pyridine, pyrimidine, possibly substituted C1-12alkylen

The invention relates to novel vitamin D analogues of the formula (I), where X is hydrogen, hydroxy or protected hydroxy; R1and R2is hydrogen, methyl or ethyl; Q3-C6hydrocarbide where hydrocarbide means biradical obtained after removal of two hydrogen atoms from a straight or branched, saturated or unsaturated hydrocarbon, in which any one of CH2group can be optionally replaced by an oxygen atom or carbonyl group such that the carbon atom (C-22), directly associated with the C-20 was a hybridized carbon atom is sp2or sp3i.e

The invention relates to novel vitamin D analogues of formula I, where X Is H or HE, R1and R2- CH3or2H5or together with the carbon atom bearing the group X, can form WITH3-C5carbocycle, Q is a single bond or C1-C8-hydrocarbide, in which one of the groups-CH2- not associated with a group, optionally substituted by an IT group or replaced by an oxygen atom, Y is a single bond or C1-C8-hydrocarbide, or other derivative

The invention relates to vitamin D analogues of General formula I, where X is hydroxy; R1and R2- the same or different, H, CH3WITH2H5or cyclopropyl; Q is methylene, ethylene, tri -, or tetramethylene, optionally substituted hydroxy-group or group-or SIG3where R3Is h, methyl or ethyl; Y is a single bond or C1-C2- hydroxycarbonyl

The invention relates to new derivatives of vitamin D3formula (I), where Q is methylene, ethylene, tri -, or tetramethylene, -CH=CH-, -CH=CH-CH=CH-; Y is a single bond, - CH(OH), -O-(C6H4)-(meta) or-S-(C6H4)-(meta); R1and R2the same or different, is methyl, ethyl, or R1and R2together with the carbon atom marked with an asterisk in formula I bearing a group Z may form WITH3-C6carbocyclization ring; Z is hydrogen or hydroxy, provided that when Y is CH(OH)-, R1and R2- mately, Z is hydroxy, the configuration at C-20 cannot be R

The invention relates to vitamin D analogues of the formula I in which Q represents a C1-C8hydrocarbononly biradical; R represents a C1-C6hydrocarbide; expression hydrocarbide (hydrocarbide) denotes the radical (biradical) obtained after removal of the 1 (2) atom(s) of hydrogen from an unbranched, branched or cyclic, saturated or unsaturated hydrocarbon, or in which two groups of R, taken together with the carbon atom carrying the hydroxyl group can form WITH3-C8carbocyclic ring

The invention relates to a synthetic derivative of vitamin D General formula I, where Q is CH2- or-CC - and-C1-C6alkylene; R1-H, C1-C4alkyl group YR1where Y -,- SO-, C1-C4alkyl, Z is H or a hydroxyl

The invention relates to veterinary

The invention relates to the field of medicine

The invention relates to a peptide selected from the group having the formula: X-R1-R2-Asp-Ala-R5-R6-Thr-R8-R9-R10-Arg-R12-R13-R14-R15-R16-Leu-R18-R19-Arg-R21-R22-Leu-Gln-Asp-Ile-R27-R28-R29-NH2where X means PhAc, IndAc or Nac, R1means Tyr or His, R2mean D-Arg, R5means Ile or Val, R6means Phe or Phe(Cl), R8means Asn, Gln, Ala or D-Asn, R9means Arg, Har, Lys, Orn, D-Arg, D-Har, D-Lys, D-Orn, Cit, Nle, Tyr(Me), Ser, Ala or Aib, R10means Tyr or Tyr(Me), R12means Lys, R13mean Val or Nle, R14means Leu or Nle, R15means Gly, Ala, Abu, Nle, or Gln, R16means Gln or Arg, R18means Ser or Nle, R19means Ala, R21means Lys, R22means Leu, Ala or Aib, R27means Met, Leu, Nle, Abu, or D-Arg, R28means Arg, D-Arg or Ser, R29means Arg, D-Arg, Har or D-Har, provided that when9and R28mean Ser, R29is Arg or Har, and its pharmaceutically acceptable salts

The invention relates to medicine, namely to Oncology, and can be used in various types of cancer of the pancreas

The invention relates to medicine, namely to dermatology, and relates to a method of treatment of mycosis fungoides
The invention relates to medicine and can be used for head and neck tumors, biliodigestive zone, rectum

The invention relates to palletization derived bacteriochlorophyll formula I, I' or I"

where a is a HE, OR1, -O-(CH2)n-Y, -S-(CH2)n-Y, -NH-(CH2)n-Y, -O-(CH2)2-OH, -NH-(CH2)2-NH-BOC, or-N(CH2-CH=CH2)2where R1represents Na+, K+, (CA2+)0,5, (Mg2+)0,5Li+, NH+4,+NH3-C(CH2OH)3,+NH3-CH2-(CHOH)4-CH2OH,+NH2(CH3)-CH2(SNON)4-CH2OH, or+N(Cn'H2n'+1)4; R2represents N or C1-C12the alkyl for the compounds of formula I', and R2represents H, HE or COOR4for the compounds of formula I, where R4represents C1-C12alkyl or C3-C12cycloalkyl; R3represents N or C1-C12the alkyl for the compounds of formula I', and R3represents H, HE, or C1-C12alkyl or alkoxy with the R'3X-where R'1, R'2and R'3each independently represents-CH3or-C2H5; X represents F, Cl, Br or I, n is 1, 2, 3 or 4, and where * denotes an asymmetric carbon atom and --- represents a single saturated bond or an unsaturated double bond pharmaceutical composition having a capacity of detection or treatment of tumors containing at least one compound of formula I, I' or I", three methods for obtaining the compounds of formula I

The invention relates to derived-alanine General formula I:

in which Q1or Q3independently from each other represent CH or N, they cannot mean N; R1represents H, A, Ar or Hal, R2denotes H or A,

each of R4and R5independently denotes H, a or Hal, R6represents H or a, a denotes alkyl with 1-6 carbon atoms, Ar is an unsubstituted aryl, Hal denotes F, Cl, Br or I, n is 2, 3, 4, 5 or 6, m is 1, 2, 3 or 4, as well as their physiologically acceptable salt and solvate; two methods for their preparation and pharmaceutical product having the properties of integrins inhibitor

The invention relates to medicine, to methods of combination therapy for cancer patients
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