RU2603959C1 - {3-[4-(7H-PYRROLO[2,3-d]PYRIMIDIN-4-YL)PYRAZOLE-1-YL]-1-ETHYLSULFONYL-AZETIDINE-3-YL}-ACETONITRILE DICHLOROACETATE AS JANUS KINASE INHIBITOR - Google Patents

Abstract

FIELD: pharmaceutics.

SUBSTANCE: invention relates to a novel salt - dichloroacetate {3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)pyrazole-1-yl]-1-ethylsulfonyl-azetidine-3-yl}-acetonitrile, which is Janus kinase inhibitor (JAK), wherein it exhibits selective action on JAK3. Compounds can be used for treating autoimmune disease or disorder, such as rheumatoid arthritis, psoriatic arthritis or juvenile arthritis. {3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)pyrazole-1-yl]-1-ethylsulfonyl-azetidine-3-yl}-acetonitrile dichloroacetate has formula (1):

.

Compound of formula 1 can be obtained by reaction of {3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)pyrazole-1-yl]-1-ethylsulfonyl-azetidine-3-yl}-acetonitrile with dichloroacetic acid.

EFFECT: invention also relates to pharmaceutical composition effective for treating JAK-related diseases in form of tablets, capsules or injections, placed in pharmaceutically acceptable pack, and method of treating.

9 cl, 2 tbl, 6 ex

Description

The present invention relates to dichloroacetate of substituted {3 - [(7H-pyrrolo [2,3-d] pyrimidin-4-yl) -azolyl] azetidine, as well as its compositions, methods of use and preparation. This compound and its compositions are inhibitors of Janus kinases (JAK) and can be used in the treatment of JAK-associated diseases, including, for example, inflammatory and autoimmune disorders, as well as cancer.

Oncogenic protein kinases are one of the largest and most attractive groups of protein targets for drug development. Janus kinases play an important role in the cytokine-dependent regulation of proliferation and function of cells involved in the immune response. Currently, four members of the JAK family in mammals are known: JAK1, JAK2, JAK3 (also known as leukocyte Janus kinase; JAKL; L-JAK) and TYK2 (also known as protein tyrosine kinase 2). Blocking signal transmission at the level of JAK kinases is promising for the development of methods for treating inflammatory diseases, autoimmune diseases, myeloproliferative and human cancers. Inhibition of JAK kinases is believed to have a therapeutic effect in patients suffering from immune disorders of the skin, such as psoriasis and skin sensitization. Effective JAK kinase inhibitors are of undoubted interest in the development of new drugs. Some JAK inhibitors, including pyrrolopyrimidines, are presented in WO 2009/114512. In this series of compounds, Baricitinib is the most advanced drug candidate. Baricitinib showed fast (after 6 months of treatment) and sustained efficacy in the treatment of rheumatoid arthritis in three phase III clinical trials [NCT02340104, NCT02263911, NCT01710358. http://www.medpagetoday.com/MeetingCoverage/EULAR/52084.http://www.fiercepharma.com/press-releases/lilly-and-incyte-announce-positive-top-line-results-phase-3- trial-baricitin].

The search for new improved JAK inhibitors is one of the main directions for developing new, more effective drugs for the treatment of rheumatoid arthritis, cancer and other diseases. The compounds and methods described herein are intended to meet these needs.

The following are definitions of terms that are used in the description of this invention.

"Active component" (drug substance, drug substance, drug-substance) means a physiologically active substance of synthetic or other (biotechnological, plant, animal, microbial and other) origin, having pharmacological activity and is the active principle of the pharmaceutical composition used for the manufacture and manufacture medicinal product (means).

“Medicinal product (preparation)” - a substance (or a mixture of substances in the form of a pharmaceutical composition) in the form of tablets, capsules, injections, ointments and other formulations intended to restore, correct or alter physiological functions in humans and animals, as well as for treatment and disease prevention, diagnosis, anesthesia, contraception, cosmetology and more.

"Pharmaceutical composition" means a composition comprising an active component and at least one of the components selected from the group consisting of pharmaceutically acceptable and pharmacologically compatible excipients, solvents, diluents, carriers, excipients, distributing and perceiving means, delivery vehicles such as preservatives, stabilizers, fillers, grinders, moisturizers, emulsifiers, suspending agents, thickeners, sweeteners, perfumes, flavors, antibacterial Gent, fungicides, lubricants, and prolonged delivery controllers, the choice and suitable proportions of which depend on the nature and way of administration and dosage. Examples of suspending agents are ethoxylated isostearyl alcohol, polyoxyethylene, sorbitol and sorbitol ether, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar-agar and tragacanth, as well as mixtures of these substances. Protection against the action of microorganisms can be achieved using a variety of antibacterial and antifungal agents, for example, such as parabens, chlorobutanol, sorbic acid and the like. The composition may also include isotonic agents, for example, sugars, sodium chloride and the like. The prolonged action of the composition can be achieved using agents that slow down the absorption of the active principle, for example aluminum monostearate and gelatin. Examples of suitable carriers, solvents, diluents and delivery vehicles are water, ethanol, polyalcohols, and also mixtures thereof, vegetable oils (such as olive oil) and injectable organic esters (such as ethyl oleate). Examples of excipients are lactose, milk sugar, sodium citrate, calcium carbonate, calcium phosphate and the like. Examples of grinders and distributors are starch, alginic acid and its salts, silicates. Examples of lubricants are magnesium stearate, sodium lauryl sulfate, talc, and high molecular weight polyethylene glycol. The pharmaceutical composition for oral, sublingual, transdermal, intramuscular, intravenous, subcutaneous, local or rectal administration of the active principle, alone or in combination with another active principle, can be administered to animals and humans in a standard administration form in the form of a mixture with traditional pharmaceutical carriers. Suitable unit dosage forms include oral forms such as tablets, gelatine capsules, pills, powders, granules, chewing gums and oral solutions or suspensions, sublingual and buccal administration forms, aerosols, implants, topical, transdermal such as ointments and creams, subcutaneous , intramuscular, intravenous, intranasal or intraocular administration forms and rectal administration forms.

An object of the present invention is to provide a novel JAK inhibitor for the treatment of rheumatoid arthritis, cancer and other diseases.

This goal is achieved by the dichloroacetate of {3- [4- (7H-pyrrolo [2,3-d] pyrimidin-4-yl) pyrazol-1-yl] -1-ethylsulfonyl-azetidin-3-yl} -acetonitrile of the formula 1 or its solvate.

The subject of the present invention is a new JAK inhibitor, which is the dichloroacetate of {3- [4- (7H-pyrrolo [2,3-d] pyrimidin-4-yl) -pyrazol-1-yl] -1-ethylsulfonyl-azetidin-3 -yl} -acetonitrile (1) or a solvate thereof.

The compounds of the present invention may also include all isotopes of atoms present in the intermediate or final compounds. Isotopes include atoms having the same atomic number but different mass numbers. For example, hydrogen isotopes include tritium and deuterium.

The new compound of formula (1) has a significantly higher solubility in aqueous media than Baricitinib, which is its undoubted advantage. Thus, the solubility of the compound of formula (1) in water at pH7 (7.10 mg / ml) is 177 times higher than the solubility of Baricitinib (0.004 mg / ml). The solubility of compound (1) at pH 2 is also higher (7.3 times) than the solubility of Baricitinib (Table 1).

The inhibitory activity of the compound of formula (1) with respect to JAK1 and JAK2 is slightly higher than the corresponding activity of Baricitinib, and with respect to JAK3 the inhibitory activity of the compound of formula (1) is slightly lower than the corresponding activity of Baricitinib. As a result, the selectivity of the compound of formula (1) with respect to JAK3 is approximately two times higher than the selectivity of Baricitinib (table 2), which is very important for JAK1 and JAK2 inhibitors. The inhibitory activity of the compounds of formula (1) and Baricitinib, shown in table 2, was obtained as a result of parallel studies under identical conditions. As a prototype, Baricitinib was selected, which is the most advanced JAK inhibitor and is currently undergoing phase III clinical trials [NCT02340104, NCT02263911, NCT01710358; J.D. Clark, M.E. Flanagan, J.-B. Telliez. Discovery and Development of Janus Kinase (JAK) Inhibitors for Inflammatory Diseases. J. Med. Chem. 2014, 57, 5023-5038].

The present invention relates to a process for the preparation of {3- [4- (7H-pyrrolo [2,3-d] pyrimidin-4-yl) pyrazol-1-yl] -1-ethylsulfonyl-azetidin-3-yl} -acetonitrile dichloroacetate (1) the interaction of {3- [4- (7H-pyrrolo [2,3-d] pyrimidin-4-yl) -pyrazol-1-yl] -1-ethylsulfonyl-azetidin-3-yl} -acetonitrile with dichloroacetic acid .

The compound of formula (1) can be obtained in accordance with the synthetic protocol described below in the examples section.

The {3- [4- (7H-pyrrolo [2,3-d] pyrimidin-4-yl) pyrazol-1-yl] -1-ethylsulfonyl-azetidin-3-yl} -acetonitrile dichloroacetate (1) of the present invention inhibits JAK activity. The term "inhibit" means the ability to reduce the activity of one or more members of the JAK family. Accordingly, a compound of formula (1) can be used in methods of inhibiting JAK activity.

In accordance with the present invention, the compound of formula (1) can act as an inhibitor of one or more JAKs.

According to the present invention, a compound of formula (1) can be used to inhibit the activity of JAK kinases in a patient who needs such modulation of the receptor by administering an inhibitory amount of a compound of formula (1).

An object of the present invention is a method of treating diseases or disorders in patients associated with JAK by administering to a patient in need of such treatment a therapeutically effective amount of a compound of the present invention or a pharmaceutical composition thereof.

A JAK-related disease can include any disease, disorder, or condition directly or indirectly associated with JAK expression or activity, including overexpression and / or an abnormal level of activity. A JAK concomitant disease may also include any disease, disorder or condition that can be prevented, ameliorated or cured by modulating JAK activity.

The subject of the present invention is a method for treating JAK-associated autoimmune diseases such as multiple sclerosis, rheumatoid arthritis, juvenile arthritis, psoriatic arthritis, type I diabetes, lupus, psoriasis, inflammatory bowel disease, ulcerative colitis, Crohn’s disease, myasthenia gravis, immunoglobulin nephropathy, and immunoglobulins, thyroid disease and the like.

According to the present invention, a compound of formula (1) can be administered to a patient in the form of pharmaceutical compositions. These compositions can be obtained by a method well known in the pharmaceutical field, and can be administered in various ways depending on whether local or systemic treatment is required and on the area to be treated. The compositions may be in the form of tablets, pills, powders, lozenges, wafers, elixirs, suspensions, emulsions, solutions, syrups, aerosols (as a solid or in a liquid medium), ointments containing, for example, up to 10% by weight of the active compound, soft and hard gelatin capsules, suppositories, sterile injectable solutions and sterile packaged powders. The compositions may be formulated in unit dosage form, each dose containing from about 5 mg to about 1000 mg, usually from about 100 mg to about 500 mg, of the active ingredient. The term "unit dose" refers to physically discrete units suitable as unit doses for humans and other mammals, each unit containing a predetermined amount of active material, calculated to produce the desired therapeutic effect, in combination with a suitable pharmaceutical excipient.

Below the invention will be described in more detail using specific examples, which are presented for purposes of illustration and are not intended to limit the invention in any way. Specialists in the art will easily understand the difference in non-critical parameters that can be changed or modified to obtain the same results.

Example 1. Dichloroacetate {3- [4- (7H-pyrrolo [2,3-d] pyrimidin-4-yl) -pyrazol-1-yl] -1-ethylsulfonyl-azetidin-3-yl} -acetonitrile (1) . To a solution of 133 mg (0.36 mmol) {3- [4- (7H-pyrrolo [2,3-d] pyrimidin-4-yl) -pyrazol-1-yl] -1-ethylsulfonyl-azetidin-3-yl} - of acetonitrile in 10 ml of tetrahydrofuran, 46.2 mg (0.36 mmol) of dichloroacetic acid are added. The mixture was evaporated to dryness on a rotary evaporator. The product is obtained as a white solid. ¹ H-NMR (DMSO-d ₆ , 400 MHz): 1.24 (t, J = 7.2Hz, 3H), 3.19-3.28 (m, 2H), 3.69 (s, 2H), 4.23 (d, J = 8.8Hz , 2H), 4.6 (d, J = 8.8Hz, 2H), 6.68 (s, 1H), 7.06-7.11 (m, 1H), 7.6-7.65 (m, 1H), 8.48 (s, 1H), 8.71 ( s, 1H), 8.93 (s, 1H), 12.16 (br.s, 1H).

Example 2. Determination of thermodynamic solubility of the compounds of formula (1) and Baricitinib. 5 mg of the test compound was mixed with 1 ml of universal buffer (pION) with pH 2.0 or pH 7.0 for 15 min at 25 ° С. Additional amounts of substances were added until the solution became cloudy. Vials with solution were incubated with stirring for 24 hours at 25 ° C to achieve equilibrium between solution and precipitate at saturation. After equilibration, 200 μl of the solution (in 2 repetitions) was filtered through a 96-well filter plate (Millipore) to separate the precipitate. The concentration of compounds in the filtrate was determined spectrophotometrically using a standard calibration curve. The optical absorption spectrum of the substance was measured and a calibration curve was constructed at the selected wavelength (usually corresponding to the maximum absorption of the substance λmax). The concentration of the substance in the filtrate (i.e. solubility) was calculated by the formula below:

Where

OD _λmax filtrate is the optical density of the filtrate,

OD _λmax blank - optical density of a blank solution without a substance,

Slope - slope of the calibration curve,

1.67 - dilution factor of the filtrate with acetonitrile,

Filtrate dilution - factor dilution of the filtrate with a buffer.

The results are presented in table 2.

Example 3. Compounds were tested in accordance with Life Technologi's standard Lyte screening protocol. Test compounds are tested in 1% DMSO (final) in the well. 100 nl 100x test compounds in 100% DMSO add 2.4 μl kinase buffer (50 mM HEPES, pH 6.5, 0.01% BRIJ-35, 10 mM MgCl ₂ , 1 mM EGTA, 0.02% NaN ₃ ) in conditions of low volume NBS, black 384 - Well plate (Corning Cat. # 4514). After that, 5 μl of the 2X peptide / kinase mixture (Tyr06 / JAK1, final concentrations of 21.2-91.5 ng JAK1 and 2 μm Tyr 06) and 2.5 μl of 4 × ATP (final concentration of 75 μM) are added to the kinase buffer. After 30 seconds on a shaker, the kinase reaction was incubated for 60 min at room temperature. After that, 5 μl in 1: 128 dilution of the development of reagent A was incubated for another 60 min at room temperature. Fluorescence was measured upon excitation at 400 nm and emission at 445 nm and 520. The degree of phosphorylation of the FRET peptide can be calculated from the emission ratio. The emission factor will remain low if the Lada peptide is not phosphorylated (i.e., not kinase inhibition) and will be high if the Lada peptide is non-phosphorylated (i.e., kinase inhibition),% phosphorylation is calculated using the formula shown on page 6 in Z′-LYTE standard screening protocol (Life Technologies).

The concentration curve of the kinase activity versus the concentration of the tested substances was constructed using the sigmoid model.

The activity data of the compounds of general formula 1 in relation to JAK1 are presented in table 2.

Example 4. Obtaining a drug in the form of tablets. 1600 mg of starch, 1600 mg of crushed lactose, 400 mg of talc and 1000 mg of inhibitor (1) were mixed and pressed into a block. The resulting bar is crushed into granules and sieved through sieves, collecting granules with a size of 14-16 mesh. The granules obtained were tabletted into tablets of suitable form weighing 500 mg each.

Example 5. Obtaining a drug in capsule form. The inhibitor (1) was thoroughly mixed with lactose in a ratio of 2: 1. The resulting powdery mixture was packaged in 600 mg in a suitable size gelatin capsule.

Example 6. Obtaining a medicinal product in the form of compositions for intramuscular, intraperitoneal or subcutaneous injection. 500 mg of inhibitor (1), 300 mg of chlorobutanol, 2 ml of propylene glycol and 100 ml of water for injection were mixed. The solution was filtered and placed in 1 ml ampoules, which were sealed.

From the above description, various modifications of the invention are apparent to those skilled in the art in addition to those described herein.

Claims (9)

1. Dichloroacetate {3- [4- (7H-pyrrolo [2,3-d] pyrimidin-4-yl) pyrazol-1-yl] -1-ethylsulfonyl-azetidin-3-yl} -acetonitrile of the formula (1)

2. An active component having the properties of a JAK inhibitor for the preparation of pharmaceutical compositions and dosage forms, which is a compound of formula (1) according to claim 1. 3. A drug having the properties of a JAK inhibitor, which is a compound of formula (1) according to claim 1. 4. A pharmaceutical composition suitable for the treatment of JAK-related diseases, comprising, as an active ingredient, a compound of formula (1) in a therapeutically effective amount and a pharmaceutically acceptable carrier and / or solvent. 5. The pharmaceutical composition according to claim 4 in the form of tablets, capsules or injections, placed in a pharmaceutically acceptable package. 6. A method of treating JAK-related diseases or disorders, comprising administering a therapeutically effective amount of a compound of formula (1) according to claim 1, or an active component according to claim 2, or a drug according to claim 3, or a pharmaceutical composition according to claim 4 or 5. 7. The method of claim 6, wherein the JAK-related disease or disorder is an autoimmune disease. 8. The method of claim 6 or 7, wherein the autoimmune disease is rheumatoid arthritis, psoriatic arthritis, or juvenile arthritis. 9. A method of obtaining a dichloroacetate of {3- [4- (7H-pyrrolo [2,3-d] pyrimidin-4-yl) pyrazol-1-yl] -1-ethylsulfonyl-azetidin-3-yl} -acetonitrile of the formula (1 ) according to claim 1, by the interaction of {3- [4- (7H-pyrrolo [2,3-d] pyrimidin-4-yl) pyrazol-1-yl] -1-ethylsulfonyl-azetidin-3-yl} -acetonitrile with dichloroacetic acid.