EA040999B1 - 4-[3-CHLORO-4-(CYCLOPROPYLCARBAMOYLAMINO)PHENOXY]-7-METHOXYQUINOLINE-6-CARBOXAMIDE METHANSULFONATE Q CRYSTAL FORM, METHOD FOR ITS PRODUCTION AND PHARMACEUTICAL COMPOSITION FOR CANCER TREATMENT - Google Patents

Description

The field of technology to which the invention belongs

The invention relates to a new, previously unknown, crystalline form Q of 4-[3-chloro-4(cyclopropylcarbamoylamino)phenoxy]-7-methoxyquinoline-6-carboxamide methanesulfonate (lenvatinib mesylate), an antitumor drug from the group of kinase inhibitors.

Background Art Information

The present invention relates to a new crystalline form Q of 4-[3-chloro-4(cyclopropylcarbamoylamino)phenoxy]-7-methoxyquinoline-6-carboxamide methanesulfonate (lenvatinib mesylate; lenvatinib methanesulfonate; INN - lenvatinib), which is characterized by the following structural formula:

Empirical formula: C21H ₁₉ ClN ₄ O ₄ -CH ₄ O ₃ S.

Molecular weight 522.96 (mesylate), 426.86 (base).

Lenvatinib is an oral multi-receptor tyrosine kinase inhibitor active against the vascular endothelial growth factor (VEGF) receptors VEGFR1 (FLT1), VEGFR2 (KDR), and VEGFR3 (FLT4). It also inhibits other kinases involved in pathological angiogenesis, tumor growth and progression, such as fibroblast growth factor receptors, FGFR1, 2, 3, and 4, as well as platelet-derived growth factor alpha receptors, such as KIT and RET. In 2015, the US Food and Drug Administration approved the use of lenvatinib for the treatment of patients with locally relapsing or metastatic, progressive, radioactive iodine-resistant thyroid carcinoma.

For the first time, crystalline forms of various lenvatinib salts are described in the patent family with a priority date of December 25, 2003 under the application JP2003-430939, including RF patent No. 2328489 C2 (publ. /10, A61R 17/06, A61R 27/02, A61R 29/00, A61R 35/00). Crystal form A was obtained by mixing lenvatinib base, a suitable solvent (methanol, ethanol) and methanesulfonic acid, followed by heating and cooling the mixture. In another variant, lenvatinib base, acetic and methanesulfonic acids were mixed, heated until dissolved, then the solvent (methanol, ethanol) was added and the mixture was slowly cooled until the corresponding crystals precipitated. Crystal form B of lenvatinib methanesulfonate was obtained by drying with aeration of the solvate of lenvatinib mesylate with acetic acid. Crystalline Form C was prepared by heating lenvatinib mesylate solvate with dimethyl sulfoxide followed by slow cooling to room temperature. In this method, the use of a solvent is optional. All of the above crystalline forms were characterized by differential scanning calorimetry and powder x-ray diffraction (see table).

U.S. Patent No. 10246418 B2 (publ. 04/02/2019, IPC: A61K 31/47, A61P 35/00, C07D 215/48, C07C 255/03) describes a method for obtaining crystalline form M of lenvatinib mesylate. Methanesulfonic acid was added dropwise to a suspension of lenvatinib base in acetonitrile and allowed to stir. The precipitate that formed was the crystalline form M of lenvatinib mesylate. The patent contains data from powder X-ray diffraction (see table), differential scanning calorimetry, thermogravimetric analysis, and IR spectroscopy. The greater solubility of the crystalline form M in comparison with the form C in water and physiological media was also demonstrated.

The method for obtaining crystalline form 1 is disclosed in the international publication WO 2018196687 A1 (publ. 11/01/2018, IPC: A61K 31/47; A61P 35/00; C07D 215/48). Example 1 describes the preparation of lenvatinib mesylate from the corresponding base in acetonitrile or acetonitrile-water in the presence of methanesulfonic acid. Example 2 shows the possibility of rapid transformation of the known crystalline form M under high humidity conditions into the claimed form 1.

International publication WO 2018122780 A1 (publ. 07/05/2018, IPC: A61K 31/47; C07D 215/00; C07D 215/48) describes the crystalline form of VN1 and the amorphous form of lenvatinib mesylate. To obtain the VN1 form, methanesulfonic acid was added to lenvatinib (as base) in methyl isobutyl ketone and stirred for 24 h. The product was isolated by filtration. The amorphous form was obtained by spray drying from a solution of lenvatinib mesylate in methanol. The claimed forms are characterized by X-ray powder diffraction and differential scanning calorimetry data.

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International patent publication WO 2019228485 A1 (publ. 05.12.2019, IPC: A61K 31/47; A61P 35/00; C07D 215/48) discloses a method for obtaining crystalline form X of lenvatinib mesylate, which includes preparing a suspension of lenvatinib base in aqueous methanol, adding methanesulfonic acid (dropwise), heating until dissolved and subsequent crystallization of the product from a solution cooled to room temperature.

According to the description of the Chinese patent application CN 110229103 A (publ. 09/13/2019, IPC: C07C 303/32; C07C 309/04; C07D 215/48), the crystalline form B of lenvatinib mesylate can be obtained from the base of lenvatinib and methanesulfonic acid in the environment isopropanol, n-propanol, tetrahydrofuran and methyl ethyl ketone. In the patent application, the powder diffraction pattern is given without indicating the numerical values of the angles 2θ. Powder diffraction patterns in the same form for other crystalline forms of lenvatinib mesylate are contained in Chinese patent applications CN 110256341 A and CN 111689897 A.

EN 2328489 С2 EP 3287444 A1 (US10246418B2) WO 2018196687 A1 WO 2019228485 A1 Form Form Form Form M Form I Form 7 Shape X A IN WITH 7.21 6.54 5.72 11.30 7.40 5.30 22.84 8.25 9.66 9.64 6.10 14.80 9.60 20.16 8.93 10.64 10.14 15.20 24.80 10.70 9.46 9.20 11.38 10.50 17.90 26.60 16.80 14.52 9.91 12.22 11.32 23.50 20.40 18.50 24.86 10.43 12.64 11.48 21.80 25.80 19.30 18.92 10.93 13.10 13.26 7.90 9.70 20.80 26.82 12.24 14.48 13.84 10.10 22.80 22.40 8.68 13.72 15.02 15.28 22.80 23.30 24.30 10.32 15.09 15.42 15.62 15.37 16.74 16.44 15.70 17.02 17.06 16.55 17.30 17.62 18.58 17.70 19.16 19.23 18.38 19.80 19.93 18.88 20.34 20.33 19.40 20.76 20.97 19.96 21.46 22.01 20.34 22.08 22.41 20.82 22.55 22.97 21.38 23.14 23.44 22.18 23.84 24.11 22.90 24.94 24.54 23.18 25.78 24.99 23.42 26.80 25.52 24.08 28.30 25.79 24.82 29.90 26.28 25.48 31.04 26.88 25.88 31.16 27.40 26.40 32.76 27.71 26.74 33.56 28.01 27.00 34.44 28.55 27.64 28.86 28.32 29.40 28.60 30.49 29.22 30.88 29.68 31.28 29.96

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31.76 30.30 32.10 31.80 32.92 32.66 33.12 32.94 33.71 33.36 34.29 35.40 34.64 36.55 34.94 37.24 36.08 38.32 36.73 38.70 37.60 38.14 38.60 39.40

CN 110903239A CN 110563644A CN 109988112A EP 3620453 Al CN 1115743 59A Form 6 4.16 4.19 8.00 5.70 9.30 5.73 10.15 11.20 6.30 11.20 6.20 12.40 12.60 8.00 13.40 8.20 16.54 16.10 11.70 13.90 10.13 18.93 17.20 12.70 20.60 10.93 20.23 17.80 15.30 22.80 11.27 22.05 18.10 16.00 24.10 11.51 23.49 18.80 17.00 25.20 12.20 26.07 19.70 19.10 26.40 12.54 29.26 20.20 19.60 26.60 13.52 20.60 20.20 28.20 14.88 21.90 20.40 16.40 22.10 23.20 17.68 22.90 24.10 18.35 23.30 25.10 19.15 23.70 25.50 19.86 24.20 26.30 20.50 25.00 27.50 21.01 25.40 29.10 21.59 26.30 22.21 27.00 22.58 28.30 23.10 28.80 23.92 29.40 24.80 25.33 26.29 27.10 27.97 29.76 31.60 33.43 34.42 35.76 37.34 38.34 39.18

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WO 2018051792 A1 WO 2019092625 A1 (lenvatinib base) Form Form Form Form Form Form Form DMSO- DMSO- ASA-1 ASA-1 CHF-1 FOA-1 H2O-1 1 2 HT DRY 4.54 4.58 5.97 6.03 4.53 4.71 4.52 4.13 9.02 9.21 7.72 7.75 9.08 9.43 5.12 10.38 15.49 12.55 10.06 11.18 10.45 9.74 6.05 12.41 17.26 13.89 10.90 12.21 13.60 10.55 9.06 14.04 18.21 15.65 11.98 13.65 15.23 11.04 10.49 16.59 18.71 18.57 12.59 15.06 15.74 11.92 14.08 17.36 19.90 19.22 13.73 15.67 16.90 14.16 16.51 19.06 20.51 21.09 15.61 16.90 17.27 14.86 19.86 20.03 22.83 22.10 16.89 17.83 18.18 15.63 24.01 20.92 25.21 23.19 17.16 18.97 18.89 16.13 26.90 21.70 27.40 25.88 17.85 19.45 19.87 16.78 22.41 29.46 26.87 19.38 20.04 20.47 17.55 23.53 32.29 31.71 20.34 21.44 21.19 18.70 26.75 36.25 37.54 20.85 23.08 21.80 19.56 28.04 38.66 21.45 23.69 22.34 20.28 29.20 22.02 24.96 22.79 20.85 29.44 22.59 25.28 23.12 21.39 23.60 26.99 25.14 22.43 24.53 28.86 25.58 22.88 25.93 31.68 26.19 23.72 26.73 33.51 26.70 24.48 28.36 35.66 27.42 24.96 29.01 37.57 28.14 25.31 30.91 29.31 26.12 32.52 31.67 26.75 35.11 32.08 27.19 36.38 32.73 27.84 37.66 33.25 30.06 34.13 31.08 36.00 31.56 36.91 32.00 38.46 34.52 39.27 35.79 38.28 39.33

Lenvatinib mesylate, like many other arylureas, is known to have a tendency to absorb moisture and gel in aqueous media. It is also known that lenvatinib is prone to hydrolysis with the formation of a genotoxic degradation product. The developers failed to create a lenvatinib drug in tablet form due to the low stability of the substance under the appropriate technological conditions for its production, since this resulted in the accumulation of a genotoxic degradation product. As a result, the drug was released and registered in the form of capsules. Low-hygroscopic auxiliary ingredients were included in the composition of the Lenvima drug, which reduced moisture absorption and increased the stability of the active substance. Specifically, calcium carbonate has been used as an insoluble excipient that prevents lenvatinib from gelling. The finished drug is available in the form of hypromellose capsules, which are significantly less hygroscopic than comparable cheaper gelatin capsules. The primary packaging of the drug is Alu-Alu type blisters, which are able to provide maximum protection against atmospheric moisture and are the most expensive option.

- 4 040999 commercially available blister packs [EMA/250082/2015 - CHMP assessment report Lenvima. 2015]. International publication WO 2018185175 A1 (publ. 10/11/2018) describes a pharmaceutical composition based on lenvatinib mesylate and sodium carbonate, which has reduced degradation and gelation in an aqueous medium.

Thus, hygroscopicity, hydrolytic degradation to toxic products and gelling tendency are well known problems associated with the use of drugs containing lenvatinib mesylate. It is also obvious that the problem of hygroscopicity and stability of the active pharmaceutical substance lenvatinib mesylate (without excipients) remains not fully resolved today.

The essence of the invention

The claimed group of inventions relates to a new, previously unknown, crystalline form Q 4[3-chloro-4-(cyclopropylcarbamoylamino)phenoxy]-7-methoxyquinoline-6-carboxamide methanesulfonate (lenvatinib mesylate; lenvatinib methanesulfonate; international non-proprietary name - lenvatinib), method its preparation, a pharmaceutical composition based on it, intended for the treatment of lenvatinib-sensitive cancers.

During experiments to optimize the conditions for the crystallization purification of lenvatinib mesylate, the authors accidentally left a Teflon spatula, which was used to stir the solution, in a beaker with a solution of lenvatinib mesylate in aqueous dimethylformamide. The next day, it was unexpectedly found that the substance had crystallized in the beaker, and the crystals on the bottom and walls of the beaker differed in appearance from the crystals grown on the Teflon spatula. The authors hypothesized that surface properties may influence the nucleation of a particular polymorph. In subsequent experiments, we investigated the crystallization of lenvatinib mesylate, first in small containers made of polytetrafluoroethylene (Teflon), and then in standard glass laboratory glassware with a hydrophobic coating on the inner surface.

In a preferred embodiment of the present invention, lenvatinib mesylate crystalline form Q was obtained by precipitation from a hot solution in hot dimethylformamide with water followed by cooling. In this case, crystallization was carried out in a glass vessel with a silanized surface. Silanization refers to the modification of a surface with silicon derivatives in order to impart hydrophobic properties to it. The crystals formed from the solution were a new, previously unknown crystalline form of lenvatinib mesylate, which we called form Q. The preservation of the molecular structure of lenvatinib mesylate was confirmed by NMR spectroscopy and mass spectrometry (Fig. 1-3). The crystallization time was usually 4-7 hours. The modification of the glass surface with silicon derivatives can be carried out by any known method, for example, described in [P. Michael Conn Neuroendocrine Peptide Methodology, 2012, 820-821]. Glass treated according to this procedure became hydrophobic and was not wetted by water. When the crystallization under similar conditions was carried out in a glass vessel made of unsilanized glass, stainless steel or enameled steel vessels, the formation of the crystalline form Q did not occur.

The structure of lenvatinib mesylate was confirmed by 1H-, ^13C -spectroscopy, mass spectrometry (Fig. 1, 2, 3). The crystalline form obtained by the process of the present invention was characterized by X-ray powder diffraction data with signals at the following 2θ angles: 4.79; 9.61; 10.80; 12.44; 15.82; 17.85; 20.61; 21.05; 21.84; 23.15; 24.30; 25.25; 26.00; 26.40; 26.72; 27.27; 27.88; 28.85° ±0.1° (Fig. 4). In qualitative phase analysis by powder X-ray diffractometry, it is customary to use the allowable convergence of 2θ angles (distance between peaks) for one crystalline form of the test substance and a standard sample within ±0.2° of the 2θ angle. Therefore, an error of ±0.1° of the angle 2θ should be applied to each of the peaks in the diffraction patterns of the studied and standard samples, since convergence is usually understood as the degree of closeness to each other of independent measurement results obtained under specific regulated conditions. [OFS.1.2.1.1.0011.15 X-ray powder diffractometry], [GOST R ISO 5725-1-2002]. Comparison of the characteristic peaks in the X-ray diffraction pattern of lenvatinib mesylate crystalline Form Q (FIG. 4) with those of prior art crystalline forms of this chemical compound, listed in the table above, suggests that Form Q is new. An independent study for a single crystal made it possible to determine the following unit cell parameters: a 8.7866(3); b 36.8509(12); s 8.8497(3); α90; β 106.1079(12); γ 90; volume 2752.9 A ³ ; and establish a monoclinic syngony for the polymorph Q. The crystalline form with these unit cell parameters is also unknown in the prior art. Subsequent deciphering of the structure and refinement of the thermal and positional parameters for the salt led to an unreliability factor of 4.51%; Z-factor: Z: 4 Z': 1. The values in brackets indicate the standard (root mean square) deviation of the value.

The phase stability of the crystalline Form Q during storage was confirmed by X-ray powder diffractometry (Example 4). As a prototype of the present invention, the authors chose the crystalline form C of lenvatinib mesylate, which was obtained by a known

- 5 040999 by the method described in the patent of the Russian Federation No. 2328489 C2, example 7. The authors noticed that in the powder of the specified form C, the particles of the substance are prone to caking. At the same time, the new crystalline form Q is a crystalline powder free of this drawback. It has also been found that manual sieving of Form C ground crystals clogs the sieve cells, while similarly sized Form Q ground crystals of the present invention easily pass through the sieves. Screening of crushed crystals of form C was only possible with the use of an ultrasonic vibrating sieve. Using the same ultrasonic vibrating sieve, the sizes of the ground form Q crystals were sieved about 2.4 times faster.

Based on the new crystalline form Q and the known crystalline form C of lenvatinib mesylate, the authors developed pharmaceutical compositions in the form of capsules containing a granular composition (granulate). During the production of the finished dosage form, the flowability of the granulate was measured using a GTB tester (ERWEKA, Germany). Flowability is most often characterized by the time (in seconds) required to empty a metal funnel with a cylindrical hole of a certain size, or by the rate (kilograms per second) of material flowing out of the funnel. For the lenvatinib mesylate granulate form C obtained according to example 3 of the present invention, flowability could not be measured by this method, since the mass was not flowable. The criterion of flowability can also be the angle of repose. The method for determining flowability by the angle of repose is based on measuring the angle between the horizontal plane and the generatrix of the cone, spontaneously created by the bulk material. The higher the flowability of the material, the smaller the angle of repose. For the composition based on lenvatinib mesylate according to example 1a, this parameter was 34°, which, according to the State Pharmacopoeia of the Russian Federation, is classified as a good degree of flowability. For the granulate based on lenvatinib mesylate according to example 3, the angle of repose is 47°, which corresponds to an unsatisfactory degree of flowability (example 8).

Based on the data obtained, it was concluded that the pharmaceutical composition based on the crystalline form Q of lenvatinib mesylate is much more preferable in terms of its technological properties for obtaining a pharmaceutical composition in the form of a granulate, in contrast to the known crystalline form C of the same compound. In addition, the lenvatinib mesylate crystalline Form Q granulate has been found to provide better dosing uniformity, which improves the quality of the respective finished drug product.

In order to study the pharmacokinetics and therapeutic efficacy of the new crystalline form Q of lenvatinib mesylate, in vivo animal studies were conducted. For comparison of pharmacokinetic parameters, Sprague-Dawley rats were divided into two groups, in each of which the animals were orally administered through a gastrointestinal tube a drug based on a specific crystalline form in the form of an aqueous suspension supplemented with 5% methylcellulose at a dosage of 3 mg/kg. The first group received crystalline form Q according to example 1a, the second group received the known crystalline form C of lenvatinib mesylate according to example 3. Also, a separate group of animals was injected with the active substance intravenously in the form of an aqueous solution. The study showed that when using the crystal form Q of lenvatinib mesylate according to the present invention, there is some increase in the area under the time-concentration curve, the maximum concentration and a decrease in the time to reach it. The bioavailability of crystalline form Q was 77.5%, while for crystalline form C this figure was 68.5% (example 10).

Therapeutic efficacy was studied in animals implanted in the hindquarters with FTC-236 cells derived from cervical lymph node metastases in patients with follicular thyroid cancer (Example 11). The mice were divided into three groups. The first received treatment with a drug based on lenvatinib mesylate according to example 1a (form Q), the second - according to example 3 (form C), the third - control, did not receive treatment. The drug was administered in the form of an aqueous suspension of crushed granulate based on the indicated crystalline forms, obtained according to example 6. At the end of the treatment period, the suppression of tumor growth was evaluated relative to the control group. According to the data obtained, the best result was observed in group 1 treated with crystalline form Q of lenvatinib mesylate (Fig. 6).

Thus, the technical results of the present invention are to improve the flowability of lenvatinib mesylate and granulate based on it, improve the uniformity of dosing in the encapsulated preparation, as well as increase the bioavailability and therapeutic efficacy in animal experiments for a new, previously unknown crystalline form Q compared to the known crystalline form. form C.

List of figures of drawings and other materials

To clarify the essence of the invention of the technical invention, FIGS. 1-6.

In FIG. 1 shows the 1H NMR spectrum of lenvatinib mesylate, obtained according to example 1a, recorded at a frequency of 600 MHz in deuterated dimethyl sulfoxide (DMSO-d ₆ ).

In FIG. 2 shows the ¹³ C NMR spectrum of lenvatinib mesylate prepared in Example 1a, recorded in deuterated dimethyl sulfoxide (DMSO-d ₆ ).

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In FIG. 3 shows the mass spectrum with electrospray ionization in the positive ion detection mode for lenvatinib mesylate obtained according to example 1a.

In FIG. 4 is an x-ray powder diffraction pattern of the crystalline form of lenvatinib mesylate of Example 1a.

In FIG. 5 shows a spatial representation of the lenvatinib mesylate molecule.

In FIG. 6 is a graph showing the change in tumor volume in treated groups compared to controls.

Information confirming the possibility of carrying out the invention

Physicochemical analysis of crystalline lenvatinib mesylate was carried out by nuclear magnetic spectroscopy 1H NMR, ¹³ C NMR, HPLC, GLC and X-ray phase analysis. 1H and ^13C NMR spectra were recorded in a saturated solution in deuterated dimethyl sulfoxide (DMSO-d ₆ ) on a Bruker AVANCE 600 high-resolution NMR spectrometer at operating frequencies of 600 and 600 MHz, respectively.

X-ray diffraction analysis was performed on a Bruker Quest D8 diffractometer equipped with a Photon-III detector (φ- and ω-scan) at 100 K using Mo Κα radiation. The structure was solved by a direct method using SHELXT [GM Sheldrick. SHELXT - Integrated spacegroup and crystal-structure determination. Acta Crystal. 2015, A71, 3-8; DOI: 10.1107/S2053273314026370] and refined according to F ² using SHELXL-2018. [, G.M. Sheldrick. Crystal structure refinement with SHELXL. Acta Crystal. 2015, C71, 3-8; DOI: 10.1107/S2053229614024218]. Absorption was taken into account using the real crystal shape or semi-empirically using the SADABS SADABS 2016/2 program: L. Krause, R. HerbstIrmer, GM Sheldrick, D. Stalke. Comparison of silver and molybdenum microfocus X-ray sources for singlecrystal structure determination J. Appl. Cryst. 2015, 48, 3-10. X-ray phase analysis was carried out on a Rigaku D/MAX-2500 X-ray diffractometer (Rigaku, Japan) in Cu-Κα radiation (λ=1.54056 A).

The water content was analyzed on a C20D automatic titrator, Mettler Toledo (Switzerland).

The flowability was measured on a GTB type tester for determining the flowability of powders/granulates (ERWEKA, Germany).

The raw material lenvatinib mesylate produced by Chongqing Carelife Pharmaceutical Co. was used as a raw material for obtaining a new crystalline Q-modification of lenvatinib mesylate. Ltd. (China).

Example 1a. Method for obtaining a crystalline form of lenvatinib mesylate in a silanized glass vessel.

In a glass vessel, the surface of which was silanized by a known method [PM Conn Neuroendocrine Peptide Methodology, 2012, 820-821], lenvatinib mesylate (5.0 g) was dissolved in dimethylformamide (about 200 ml) with stirring at a temperature of about +60°C. To the solution was added about 300 ml of water and cooled with stirring to 3-5°C. The precipitate formed was filtered off on a fine Schott filter, washed with water, and dried to constant weight under vacuum at a temperature of about +40°C. 4.62 g of lenvatinib mesylate crystalline form Q was obtained, characterized by X-ray diffraction analysis. The molecular structure retention of lenvatinib mesylate was confirmed by 1 H NMR spectroscopy (FIG. 1), ¹³ C NMR spectroscopy (FIG. 2) and mass spectrometry (FIG. 3). The powder diffraction pattern is shown in Fig. 4.

Example 1b. Method for obtaining a crystalline form of lenvatinib mesylate in a vessel made of polytetrafluoroethylene.

Lenvatinib mesylate crystalline Form C was prepared analogously to method 1a, using a PTFE vessel instead of a glass vessel. The Q crystalline form of lenvatinib mesylate was obtained in 83% yield. The powder diffractogram of the product is identical to the powder diagram of the sample obtained in example 1a (Fig. 4).

Example 2. Method for obtaining a crystalline form of lenvatinib mesylate (zooming).

In a glass vessel, the surface of which was silanized in a known manner [P.M. Conn Neuroendocrine Peptide Methodology, 2012, 820-821], dissolved lenvatinib mesylate (50.0 g) in about 2.0 L of dimethylformamide with stirring and heating to 60°C. To the solution was added about 3.0 l of water with stirring and cooled to 3-5°C. The precipitate formed was filtered off on a fine Schott filter and dried to constant weight under vacuum at a temperature of about +40°C. Received 45.6 g of the crystalline form Q of lenvatinib mesylate, characterized by X-ray diffraction analysis. The powder diffractogram of the product is identical to the powder diagram of the sample obtained in example 1a (Fig. 4).

Example 3. Obtaining crystalline form C of lenvatinib mesylate by a known method (prototype, patent RU 2328489 C2, example 7).

To a suspension of 4-(3-chloro-4-(cyclopropylaminocarbonyl)aminophenoxy)-7-methoxy-6quinolinecarboxamide (1.28 g) in acetic acid (12.8 ml) was added methanesulfonic acid (0.408 ml) and stirred at room temperature until complete dissolution. The solution was heated in a water bath to 30° C. and isopropyl alcohol (7.7 ml) was added. Isopropyl alcohol (18 ml) was added dropwise to the resulting solution in equal portions over 40 min. The mixture was cooled to room temperature, then to a temperature of 15°C. The precipitated crystals were filtered off, washed with isopropyl alcohol, then suspended in ethanol at room temperature for 1 h. The product was filtered off, washed with ethanol, and dried to constant weight. The powder diffraction pattern of the resulting product matched that of crystalline Form C shown in FIG. 9 drawings to the patent of the Russian Federation No. 2328489 C2.

Example 4 Phase Stability Study of Lenvatinib Mesylate Samples.

The stability of samples of crystalline Form Q of lenvatinib mesylate obtained according to examples 1a, 1b and 2 was confirmed by the absence of a shift beyond the ±0.2° confidence interval or the appearance of additional Bragg peaks in the powder X-ray diffraction spectrum after 6 months of storage at a temperature of 25±2°C and humidity 60±5%.

Example 5. Determination of water content in samples of lenvatinib mesylate by the method of K. Fischer.

The determination was performed by the micromethod in accordance with the EAEU Pharmacopoeia (2.1.5.13. Water: microdetermination) on an automatic coulometric titrator without a diaphragm, for example, 899 Coulometer (Metrohm, Switzerland) or similar. About 100.0 mg of the substance was placed in a dry vial with a capacity of 20 ml. The vial with a rubber stopper was weighed with an accuracy of ±0.0002 g, 10 ml of K. Fischer titration reagent was added and immediately tightly closed. Weighed the vial with the entered solvent and calculated the value of the sample of the solvent, in grams. The resulting solution was subjected to ultrasound at a temperature of 50°C until the sample was completely dissolved, and the solution was cooled to room temperature. The weight of the test solution was calculated and this value was entered into the instrument. The value of the water content in an aliquot of the test solution (w), calculated by the built-in software, was obtained, in parts per million (ppm). Titration was carried out until the end point of the titration was established.

The water content in the substance in the test sample (W), in percent, was calculated by the formula: W ₌ w _iX ( _ao+n )-w _o x _a| , _xl00% 10000χα where w _i is the water content in an aliquot of the test solution, in ppm;

w ₀ - water content in the solvent, in ppm;

a ₀ is the weight of the sample of the solvent, in grams;

a is the mass of the sample of the substance, in grams.

Lenvatinib mesilate by example 1a Lenvatinib mesilate by example 1b Lenvatinib mesilate by example 2 Lenvatinib mesilate by example 3 Water content, % 0.08 oh and 0.12 0.88

Example 6. Obtaining a finished drug based on crystalline lenvatinib mesylate in the form of capsules, 4 mg.

Weighed on the scales, sifted and hung in individual bags:

crystalline lenvatinib mesylate according to examples 1a or 3 - 212.0 g;

calcium carbonate - 1430.0 g;

mannitol - 697.0 g;

low-substituted hydroxypropylcellulose - 1083.0 g;

microcrystalline cellulose - 433.0 g;

hydroxypropyl cellulose - 130.0 g.

For sifting magnesium stearate, sieves with a mesh size of 0.200 ± 0.0083 mm were used, for other components - 0.400 ± 0.015 mm. The crystalline form C of lenvatinib mesylate was sieved using an ultrasonic vibrating sieve, otherwise the powders were sieved through the sieves by hand.

Crystalline lenvatinib mesylate and calcium carbonate were loaded into the granulator, mixed at a speed of 15±1 rpm for 5-7 minutes. Then mannitol, low-substituted hydroxypropyl cellulose and microcrystalline cellulose were added and mixed at a speed of 20±1 rpm for 8-9 minutes. Hydroxypropylcellulose was dissolved in purified water in a stainless steel container. After mixing, an aqueous solution of hydroxypropyl cellulose was added to the granulator for 7-8 minutes and granulated. The wet granules were dried using a fluidized bed dryer with an air temperature of 70°C inlet and 40°C outlet. The dried granules were sifted through sieves. The crushed granules, microcrystalline cellulose and talc were loaded into the mixer. The mixture was stirred for 7-9 minutes at a speed of 15±1 rpm. Using a capsule machine, the resulting granulate was encapsulated in hypromellose capsules. Capsule weight 100.0 g ±7.5%.

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Example 7. Study of the stability of the finished medicinal product based on crystalline lenvatinib mesylate.

Accelerated drug stability tests were carried out for 6 months at a temperature

40±2°C and relative humidity 75±5%. Based on the results of the stability study, the shelf life of the medicinal product was confirmed within 24 months.

Data on the stability of the drug based on lenvatinib mesylate according to example 1 a

Index Regulatory requirements 0 months 3 months 6 months 9 months 12 months Authenticity Retention time of the main peak in the chromatogram Resp. Resp. Resp. Resp. Resp. Dissolution At least 80% (Q) of the claimed amount of lenvatinib must be go into solution after 30 minutes 94.5 95.2 95.3 95.1 95.0 The amount of impurities Not more than 0.5% 0.12 0.15 0.18 0.26 0.32 quantitative definition The content in the tablet should be from 95% to 105% of the declared amount 98.1 98.1 97.9 97.8 97.4 Water Not more than 3.0% 1.4 1.6 1.7 1.7 1.8

Data on the stability of the drug based on lenvatinib mesylate according to example 3

Index Regulatory requirements 0 months 3 months 6 months 9 months 12 months Authenticity retention time Resp. Resp. Resp. Resp. Resp. main peak on chromatogram Dissolution At least 80% (Q) of the claimed amount of lenvatinib must be go into solution after 30 minutes 86.8 86.7 85.9 85.7 85.4 The amount of impurities Not more than 0.5% 0.21 0.22 0.24 0.29 0.35 quantitative definition The content in the tablet should be from 95% to 105% of the declared amount 100.0 100.1 99.9 99.7 99.6 Water Not more than 3.0% 1.7 1.9 2.1 2.2 2.4

Example 8 Measurement of the angle of repose of the granulate of lenvatinib mesylate samples.

To determine the flowability, a device was used, consisting of a base with applied concentric circles with a diameter of 100 to 500 mm every 10 mm, a rack with a scale for measuring height, a movable bar capable of moving in vertical and horizontal planes. A hollow cylinder (diameter 100 mm and height 300 mm) was placed on the base, matching it with a circumference of 100 mm. 100 g of granulate was poured into it and lifted strictly vertically upwards with the same speed in all experiments. Spilling, the material acquired a shape close to conical. The experiment was repeated 5-6 times, each time fixing the height of the cone h and its diameter D. Based on the arithmetic mean of these measurements, the angle of repose α was calculated: h α = arctg-----0.5 ^

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G ranulat lenvatinib mesylate according to example 1a G ranulat lenvatinib mesylate according to example 3 Angle of repose, deg. 34±1 47±1

Example 9. Determination of the uniformity of dosing of the finished medicinal product.

The determination was carried out by high performance liquid chromatography (HPLC) in accordance with the Pharmacopoeia of the Eurasian Economic Union (2.1.2.28. High performance liquid chromatography, 2.1.2.36 Chromatographic separation methods) on a high pressure liquid chromatograph equipped with an ultraviolet detector, for example, Agilent 1260 (Agilent Technologies, USA). The drug capsule was placed in a volumetric flask with a capacity of 50 ml, 35 ml of solvent was added, shaken on a shaker at a speed of 350 rpm for 30 min, sonicated for 15 min with occasional stirring, the solution was cooled to room temperature, the volume of the solution was adjusted with a solvent to labels and mixed.

1.5 ml of the resulting filtrate was placed in a volumetric flask with a capacity of 25 ml, the volume of the solution was brought up to the mark with the mobile phase and stirred. The procedure was repeated for nine more capsules.

The content of lenvatinib in the capsule (xi), as a percentage of the declared amount, was calculated by the formula _ SiXaoXPxKxViXVsXlOO _ Sixa ₀ xPxKxV ₁ xV ₃

Xf — Σ GT 77777 : 7 77 — Σ 77 7 77

SqXNxIOOxIxLxK,

S ₀ xNxLxV ₂ where Si is the area of the lenvatinib peak on the chromatogram of the test solution;

S ₀ - area of the peak of lenvatinib on the chromatogram of the standard solution;

and ₀ is the weight of the standard sample (RS) of lenvatinib mesylate, taken to prepare the standard solution, in mg;

P is the content of lenvatinib mesylate in the SS of lenvatinib mesylate, in percent;

K is the conversion factor of lenvatinib mesylate to lenvatinib (=0.8163);

N is the dilution factor of the standard solution, in milliliters (=10000);

V1 is the volume of the first volumetric flask for preparing the test solution (=50 for a dosage of 4 mg), in ml;

V ₂ - aliquot for the preparation of the test solution (=1.5 for a dosage of 4 mg), in ml;

V ₃ - the volume of the second volumetric flask for the preparation of the test solution (=25 for a dosage of 4 mg), in ml;

L is the declared amount of lenvatinib in the formulation (=4 for a 4 mg dosage), in mg.

Capsules with lenvatinib mesylate according to example 1a Capsules with lenvatinib mesylate according to example 3 First indicator acceptability (AV), % 4.9 15.0

Example 10 In vivo pharmacokinetic study of lenvatinib mesylate samples in rats.

Male Sprague Dawley rats aged 7 weeks weighing 180-200 g were kept under standard laboratory conditions at a temperature of 21-23°C and a humidity of 45-55%. The animals had free access to food and water. The rats were divided into groups of 12 individuals each. The first group received, orally via a gastrointestinal tube, an aqueous suspension supplemented with 5% (V/V) methylcellulose based on the Q crystalline form of lenvatinib mesylate according to example 1a, the second received a suspension supplemented with 5% (V/V) methylcellulose based on the crystalline form lenvatinib mesylate C according to example 3 (prototype). An aqueous solution based on the known crystalline form C of lenvatinib mesylate was administered intravenously. The dosage for oral and parenteral administration was 3 mg/kg.

Blood samples were taken from the catheter in the jugular vein (200 μl) through 0, 0.083 (only for intravenous administration), 0.25; 0.5; 1; 2; 4; 8; 12; 24; 48; 72 and 96 hours after drug administration. Plasma was centrifuged at room temperature, then stored at -80°C. Plasma samples were analyzed by HPLC to determine the concentration of lenvatinib mesylate.

For intravenous administration, the following pharmacokinetic parameters were obtained: AUC(o^ _inf )= 30.25 mgh/ml; C _max =14.08 mg/ml; t _1/2 \u003d 3.51 hours

The bioavailability of the oral form of administration was calculated by the formula

F= X 100%, \AUC _BB xD _x J where AUC _x is the area under the concentration-time curve after oral administration;

AUC _bb - area under the curve concentration - time after intravenous administration;

D _x - dosage of the drug for oral administration;

D _bb - dosage of the drug for intravenous administration.

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Claims (4)

Parameter Lenvatinib mesilate according to example 1a Lenvatinib mesylate according to example 3 (prototype) AUC (o^inf), mg*h/ml 23.44 20.72 Stakh, MG/ML 6.99 6.41 tl/2, h 3.01 3.48 F*, % 77.5 68.5 * differences can be considered statistically significant (p<0.05). Example 11 Therapeutic Efficacy Study of Lenvatinib Mesylate Samples. Comparison of the therapeutic efficacy of samples of lenvatinib mesylate was carried out on nude mice at the age of 5-6 weeks. Animals were injected into the back of the body with FTC-236 cells obtained from metastases of the cervical lymph node in follicular thyroid cancer in a volume of 0.1-0.2 ml at a cell density of 5-10x10 ⁷ cells/ml. Upon reaching the volume of the tumor 200-300 mm ³ animals were randomly divided into three groups, 12 individuals in each. Tumor volume was measured twice a week and calculated by the formula [(D)xd]/2, where D is the largest tumor diameter, d is the smallest. The change in tumor volume (its growth) relative to the control group was estimated by the formula ΔT/С=(ΔT/ΔС)χ100%, where ΔΤ - tumor growth in the treatment group, ΔΟ - tumor growth in the control group. Group 1 received lenvatinib mesylate treatment according to example 1a, group 2 - according to example 3, group 3 (control) did not receive treatment. The lenvatinib mesylate preparation was administered orally via a gastrointestinal tube once a day as an aqueous suspension of ground granulate based on the indicated forms for 15 days. The results were evaluated at the end of treatment. The greatest suppression of tumor growth was observed in group 1, where the treatment was performed with a drug based on the crystalline form Q of lenvatinib mesylate according to example 1a. So, in the control group, the tumor size was about 1000 mm ³ in the first group 405 mm ³ and in the second 560 mm ³ . Differences in the obtained results can be considered statistically significant, taking into account the achieved criterion p<0.05. A plot of tumor volume change (ΔΤ/C) data is shown in FIG. 5. CLAIM 1. Crystalline form Q of 4-[3-chloro-4-(cyclopropylcarbamoylamino)phenoxy]-7methoxyquinoline-6-carboxamide methanesulfonate, characterized by unit cell parameters: a 8.7866(3); b 36.8509(12); s 8.8497(3); α90; β 106.1079(12); gamma 90. 2. The crystalline form according to claim 1, further characterized by signals at 4.79; 9.61; 10.80; 12.44; 15.82; 17.85; 20.61; 21.05; 21.84; 23.15; 24.30; 25.25; 26.00; 26.40; 26.72; 27.27; 27.88; 28.85° ±0.1° of the 2Θ angle on a powder diffraction pattern obtained in Cu-Ka radiation with a wavelength of 1.54056 A. 3. The method for obtaining the crystalline form Q according to claim 1, comprising the steps of preparing a solution of 4-[3-chloro-4-(cyclopropylcarbamoylamino)phenoxy]-7-methoxyquinoline-6-carboxamide methanesulfonate in dimethylformamide in a vessel with a hydrophobic inner surface at a temperature of about 60°C, adding water, cooling with stirring to a temperature of 3-5°C, filtering and drying under vacuum at a temperature of about +40°C. 4. Pharmaceutical composition for oral administration, intended for the treatment of cancers sensitive to lenvatinib, containing as a pharmacologically active ingredient the crystalline form Q of 4-[3-chloro-4-(cyclopropylcarbamoylamino)phenoxy]-7-methoxyquinoline-6carboxamide methanesulfonate according to p. 1 in combination with excipients. -