WO2018157803A1 - Venetoclax crystal forms and preparation method therefor - Google Patents

Abstract

Venetoclax crystal forms and a preparation method therefor, the structural formula thereof is represented by formula (I), and such crystal forms may be directly obtained from crystallization in a solvent.

Description

Crystal form of virendola and preparation method thereof Technical field

The invention relates to the field of chemical medicine, in particular to a crystal form of vintomone and a preparation method thereof.

Background technique

The dynamic balance of apoptosis and proliferation is the most basic biological process for multicellular organisms to maintain their structural stability and balance of internal environment functions and growth. Studies have shown that the proto-oncogene Bcl-2 is the main cause of inhibition of apoptosis, and the Bcl-2 protein controlled by it has a function of blocking apoptosis from protozoa to human cells, and in some types of cancer. Overexpression is associated with the formation of drug resistance. Since Bcl-2 is highly expressed in cancer cells, Bcl-2 family protein inhibitors can selectively exert anti-tumor effects in tumor cells.

Veneta, also known as Venetoclax, ABT199, is a selective, potent, orally administered small molecule Bcl-2 inhibitor. On April 11, 2016, the FDA approved its marketing under the trade name Venclexta for the treatment of chronic lymphocytic leukemia (CLL). The chemical name of Venetatal is 4-(4-{[2-(4-chlorophenyl)-4,4-dimethylcyclohex-1-en-1-yl]methyl}piperazine-1 -yl)-N-({3-nitro-4-[(tetrahydro-2H-pyran-4-ylmethyl)amino]phenyl}sulfonyl)-2-(1H-pyrrolo[2, 3-b]pyridin-5-yloxy)benzamide (hereinafter referred to as "compound (I)"), and has a structural formula of the formula I.

In the field of pharmaceutical research, different drug crystal forms can have different colors, melting points, solubility, dissolution properties, chemical stability, mechanical stability, etc., which can affect the quality, safety and effectiveness of pharmaceutical preparations, leading to clinical Difference in efficacy. Therefore, crystal form research and control has become an important research content in the drug development process.

At present, a preparation method of the compound (I) is disclosed in Example 5 of CN103153993A, in which the solid of the compound (I) obtained is amorphous. CN103328474A discloses a crystalline form of compound (I) comprising 2 anhydrates A and B, 2 hydrates C and D and various solvates E, F, G, H, I, J, anhydrous in the patent Both the substance and the hydrate need to be obtained by drying the solvate, which determines that the preparation process requires a synthetic solvate intermediate, the preparation method is complicated, the operation is cumbersome, and it is not suitable for industrial large-scale production. Further, the inventors have found through research that the anhydrate B in CN103328474A is more stable than other crystal forms, but has problems such as high wettability and low solubility. Therefore, a novel anhydrate and hydrate form of the compound (I) can be provided, and these anhydrates and hydrates can be directly crystallized from a solvent, and have good stability, high solubility, low wettability, and uniform particle size distribution. It will be of great significance for the further development of the drug.

Summary of the invention

The technical problem to be solved by the present invention is to overcome the deficiencies of the prior art, and to provide an anhydrous form, a hydrate, a solvate form of the compound (I), a preparation method thereof and use thereof.

According to the object of the present invention, the present invention provides the crystal form CS1 of the compound (I) (hereinafter referred to as "crystal form CS1"). The crystal form CS1 is a hydrate.

Using Cu-Kα radiation, the X-ray powder diffraction of the crystal form CS1 has a characteristic peak at a diffraction angle 2θ of 4.8°±0.2°, 17.0°±0.2°, and 19.0°±0.2°.

Further, the X-ray powder diffraction of the crystal form CS1 has a characteristic peak at one or two or three of the diffraction angle 2θ of 8.7°±0.2°, 13.9°±0.2°, and 19.6°±0.2°. Preferably, the X-ray powder diffraction of the crystal form CS1 has a characteristic peak at a diffraction angle 2θ of 8.7°±0.2°, 13.9°±0.2°, and 19.6°±0.2°.

Further, the X-ray powder diffraction of the crystal form CS1 has a characteristic peak at one or two or three points in the diffraction angle 2θ of 6.4°±0.2°, 11.2°±0.2°, and 17.9°±0.2°. Preferably, the X-ray powder diffraction of the crystalline form CS1 has a characteristic peak at a diffraction angle 2θ of 6.4°±0.2°, 11.2°±0.2°, and 17.9°±0.2°.

Further, the X-ray powder diffraction of the crystal form CS1 has one or more of the diffraction angle 2θ of 9.7°±0.2°, 14.9°±0.2°, 21.2°±0.2°, and 22.8°±0.2°. Characteristic peaks. Preferably, the X-ray powder diffraction of the crystalline form CS1 has a characteristic peak at a diffraction angle 2θ of 9.7°±0.2°, 14.9°±0.2°, 21.2°±0.2°, and 22.8°±0.2°.

In a preferred embodiment, the X-ray powder diffraction of the crystalline form CS1 is 4.8°±0.2°, 17.0°±0.2°, 19.0°±0.2°, 8.7°±0.2°, 13.9°± at the diffraction angle 2θ. 0.2°, 19.6°±0.2°, 6.4°±0.2°, 11.2°±0.2°, 17.9°±0.2°, 9.7°±0.2°, 14.9°±0.2°, 21.2°±0.2°, 22.8°±0.2° There are characteristic peaks.

Without limitation, in one embodiment of the invention, the X-ray powder diffraction pattern of Form CS1 is shown in FIG.

According to the object of the present invention, the present invention also provides a method for preparing the crystalline form CS1, which comprises obtaining a target solvent set by steam distillation using a tetrahydrofuran solution of the compound (I), and obtaining the crystal by crystallization at a certain temperature. The target solvent is a C ₃ -C ₈ ester solvent;

among them:

Preferably, the ester solvent is ethyl formate, ethyl acetate, isopropyl acetate or a mixed solvent thereof, more preferably isopropyl acetate;

Preferably, the crystallization temperature is 20-40 ° C, more preferably room temperature;

Preferably, the crystallization time is 6-24 hours, more preferably 12 hours.

According to the object of the present invention, the present invention provides the crystal form CS2 of the compound (I) (hereinafter referred to as "crystal form CS2"). The crystalline form CS2 is a 1,4-dioxane solvate.

Using Cu-Kα radiation, the X-ray powder diffraction of the crystal form CS2 has a characteristic peak at a diffraction angle 2θ of 5.4°±0.2°, 8.0°±0.2°, and 18.7°±0.2°.

Further, the X-ray powder diffraction of the crystalline form CS2 has a characteristic peak at one or two or three of the diffraction angle 2θ of 14.5°±0.2°, 19.6°±0.2°, and 20.0°±0.2°. Preferably, the X-ray powder diffraction of the crystalline form CS2 has a characteristic peak at a diffraction angle 2θ of 14.5°±0.2°, 19.6°±0.2°, and 20.0°±0.2°.

Further, the X-ray powder diffraction of the crystalline form CS2 has a characteristic peak at one or two of the diffraction angle 2θ of 15.9°±0.2° and 18.1°±0.2°. Preferably, the X-ray powder diffraction of the crystalline form CS2 has a characteristic peak at a diffraction angle 2θ of 15.9°±0.2° and 18.1°±0.2°.

In a preferred embodiment, the X-ray powder diffraction of the crystalline form CS2 is 5.4°±0.2°, 8.0°±0.2°, 18.7°±0.2°, 14.5°±0.2°, 19.6°± at the diffraction angle 2θ. There are characteristic peaks at 0.2°, 20.0°±0.2°, 15.9°±0.2°, and 18.1°±0.2°.

Without limitation, in one embodiment of the invention, the X-ray powder diffraction pattern of Form CS2 is shown in FIG.

According to the object of the present invention, the present invention also provides a method for preparing the crystalline form CS2, which comprises obtaining a target solvent sleeve steam displacement using a tetrahydrofuran solution of the compound (I), and obtaining the crystal by crystallization at a certain temperature. The target solvent is 1,4-dioxane;

among them:

Preferably, the crystallization temperature is 20-40 ° C, more preferably room temperature;

Preferably, the crystallization time is 6-24 hours, more preferably 12 hours.

According to an object of the present invention, the present invention provides a crystal form CS3 of the compound (I) of the formula (hereinafter referred to as "crystal form CS3"). The crystal form CS3 is an anhydride.

Using Cu-Kα radiation, the X-ray powder diffraction of the crystal form CS3 has a characteristic peak at a diffraction angle 2θ of 10.4°±0.2°, 15.2°±0.2°, and 21.7°±0.2°.

Further, the X-ray powder diffraction of the crystalline form CS3 has a characteristic peak at one or two or three of the diffraction angle 2θ of 5.2°±0.2°, 19.7°±0.2°, and 29.4°±0.2°. Preferably, the X-ray powder diffraction of the crystalline form CS3 has a characteristic peak at a diffraction angle 2θ of 5.2°±0.2°, 19.7°±0.2°, and 29.4°±0.2°.

Further, the X-ray powder diffraction of the crystal form CS3 has a characteristic peak at one or two or three points in the diffraction angle 2θ of 20.9°±0.2°, 24.3°±0.2°, and 26.2°±0.2°. Preferably, the X-ray powder diffraction of the crystalline form CS3 has a characteristic peak at a diffraction angle 2θ of 20.9°±0.2°, 24.3°±0.2°, and 26.2°±0.2°.

In a preferred embodiment, the X-ray powder diffraction of the crystalline form CS3 is 10.4°±0.2°, 15.2°±0.2°, 21.7°±0.2°, 5.2°±0.2°, 19.7°± at the diffraction angle 2θ. There are characteristic peaks at 0.2°, 29.4°±0.2°, 20.9°±0.2°, 24.3°±0.2°, and 26.2°±0.2°.

Without limitation, in one embodiment of the invention, the X-ray powder diffraction pattern of Form CS3 is shown in FIG.

According to the object of the present invention, the present invention also provides a method for preparing the crystalline form CS3, which comprises: stirring and crystallization using a solution of the compound (I) in tetrahydrofuran at a certain temperature, and filtering the filter cake to be added to the tetrahydrofuran solvent again after filtration. Medium, dissolved by heating, then concentrated, and obtained by crystallization at a certain temperature;

among them:

Preferably, the crystallization temperature is 20-40 ° C, more preferably room temperature;

Preferably, the crystallization time is from 3 to 24 hours, more preferably from 3 hours.

In the preparation method of the crystal form of the present invention:

The "room temperature" means 20-30 °C.

The "stirring" is carried out by a conventional method in the art, such as magnetic stirring or mechanical stirring, and the stirring speed is 50 to 1800 rpm, preferably 300 to 900 rpm.

The "sleeve steaming" is accomplished by conventional methods in the art by first evaporating the original solvent and then adding another solvent.

According to the purpose of the present invention, the present invention provides the crystal forms CS4, CS5, CS6 of the compound (I). The crystal forms CS4, CS5, and CS6 are MIBK (methyl isobutyl ketone) solvates.

Without limitation, in a particular embodiment of the invention, the X-ray powder diffraction patterns of Forms CS4, CS5, CS6 are shown in Figures 13, 17, and 21.

The present invention provides a pharmaceutical composition comprising a therapeutically effective amount of crystalline form CS1, CS3 or a mixture thereof and a pharmaceutically acceptable pharmaceutical excipient.

Further, the present invention provides the use of the crystalline form CS1, CS3 or a mixture thereof or the pharmaceutical composition for the preparation of a medicament for the treatment of chronic lymphocytic leukemia.

In the present invention, "crystal" or "polymorph" means confirmed by the X-ray diffraction pattern characterization shown. Those skilled in the art will appreciate that the physicochemical properties discussed herein can be characterized, with experimental error depending on the conditions of the instrument, the preparation of the sample, and the purity of the sample. In particular, it is well known to those skilled in the art that the X-ray diffraction pattern will generally vary with the conditions of the instrument. It is particularly important to note that the relative intensities of the X-ray diffraction patterns may also vary with experimental conditions, so the order of peak intensities cannot be the sole or decisive factor. In fact, the relative intensity of the diffraction peaks in the XRPD pattern is related to the preferred orientation of the crystal. The peak intensities shown here are illustrative and not for absolute comparison. In addition, the experimental error of the peak angle is usually 5% or less, and the error of these angles should also be taken into account, and an error of ±0.2° is usually allowed. In addition, due to experimental factors such as sample height, the overall offset of the peak angle is caused, and a certain offset is usually allowed. Thus, it will be understood by those skilled in the art that the X-ray diffraction pattern of one crystal form in the present invention is not necessarily identical to the X-ray diffraction pattern in the example referred to herein, and the "XRPD pattern is the same" as used herein does not mean absolutely the same. The same peak position can differ by ± 0.2° and the peak intensity allows for some variability. Any crystal form having a map identical or similar to the characteristic peaks in these maps is within the scope of the present invention. One skilled in the art will be able to compare the maps listed herein with a map of an unknown crystal form to verify whether the two sets of maps reflect the same or different crystal forms.

In some embodiments, the crystalline forms CS1 and CS3 of the present invention are pure, unitary, and substantially free of any other crystalline form. In the present invention, "substantially free" when used to refer to a new crystalline form means that the crystalline form contains less than 20% by weight of other crystalline forms, especially less than 10% by weight of other crystalline forms, more Other crystal forms of 5% by weight, more preferably less than 1% by weight of other crystal forms.

It should be noted that the numerical values and numerical ranges recited in the present invention are not to be construed as narrowly construed as a numerical value or a numerical range per se. It will be understood by those skilled in the art that they may vary depending on the specific technical environment without departing from the spirit of the invention. On the basis of the principle, there are fluctuations around specific numerical values. In the present invention, such a floating range which can be foreseen by those skilled in the art is often expressed by the term "about".

The crystal forms CS1 and CS3 provided by the present invention have the following advantages compared with the prior art:

(1) The crystal form provided by the invention has good physical and chemical stability, thereby ensuring consistent and controllable quality standards of the sample, and meeting the stringent requirements for the crystal form in the pharmaceutical application and preparation process. The crystalline form CS1 of the present invention is kept at 25 ° C / 60% RH, 40 ° C / 75% RH and 60 ° C / 75% RH for at least 3 weeks, and the crystalline form CS3 is at 25 ° C / 60% RH, 40 ° C /75%RH and 60 °C / 75% RH for at least 2 weeks; the crystal form CS1 and CS3 have little change in purity before and after placement, and have good physical and chemical stability, which is conducive to sample preservation and formulation stability. .

(2) The crystal form provided by the present invention has good mechanical stability, and the crystal form remains unchanged after grinding. Good mechanical stability can reduce the risk of crystal transformation during grinding or tableting during preparation. The crystal forms CS1 and CS3 of the invention have high grinding stability, and the grinding and pulverizing of the raw material medicine are often required in the processing of the preparation, and the high grinding stability can reduce the crystallinity change and the crystal transformation of the raw material medicine during the processing of the preparation. risk.

(3) The crystal form provided by the invention has good solubility, can reduce the dosage of the drug, thereby reducing the side effects of the drug and improving the safety of the drug, and can achieve the desired therapeutic blood concentration without a high dose after oral administration. Conducive to the absorption of drugs in the human body, so as to achieve the desired bioavailability and efficacy of the drug, in line with medicinal requirements;

(4) The crystal form provided by the invention has low wettability and can overcome the disadvantages caused by high wettability, such as the weight change of the water absorption due to the weight change of the raw material, which is favorable for long-term storage of the medicine and reduction of material storage and Quality control costs. The crystal form CS3 provided by the present invention has a weight gain of 1.12% under 80% relative humidity, and has lower wettability than the prior art. The low wettability of the crystalline form CS3 of the present invention can well resist the problem of crystal form instability during the preparation of the pharmaceutical preparation and/or storage, and the unworkability of the preparation caused by external factors such as environmental moisture, and is advantageous for preparation of the preparation. Accurate quantification and later transport and storage;

(5) The crystal form CS3 particles of the present invention are normally distributed and have a narrow particle size distribution. Its uniform particle size helps to simplify the post-treatment process of the formulation process, such as reducing the grinding of the crystal, saving cost, reducing the crystallinity change and the risk of crystal transformation in the grinding, and improving the quality control. Its narrow particle size distribution can improve the uniformity of the drug substance components in the preparation, and make the difference between different batches of preparations smaller, such as more uniform dissolution; its smaller crystal grain size can increase the drug ratio. The surface area increases the dissolution rate of the drug, which is beneficial to the absorption of the drug, thereby improving the bioavailability;

In addition, the novel solvate provided by the present invention can be used in process development intermediates, and the purification effect is remarkable, which is very important for drug development.

DRAWINGS

Figure 1 is an XRPD pattern of the compound (I) crystal form CS1

Figure 2 is a ¹ H NMR chart of the compound (I) crystal form CS1

Figure 3 is a DSC chart of the compound (I) crystal form CS1

Figure 4 is a TGA diagram of the compound (I) crystal form CS1

Figure 5 is an XRPD pattern of the compound (I) crystal form CS2

Figure 6 is a ¹ H NMR chart of the compound (I) crystal form CS2

Figure 7 is a DSC chart of the compound (I) crystal form CS2

Figure 8 is a TGA diagram of the compound (I) crystal form CS2

Figure 9 is an XRPD pattern of the compound (I) crystal form CS3

Figure 10 is a ¹ H NMR chart of the compound (I) crystal form CS3

Figure 11 is a DSC chart of the compound (I) crystal form CS3

Figure 12 is a TGA diagram of the compound (I) crystal form CS3

Figure 13 is an XRPD pattern of the compound (I) crystal form CS4

Figure 14 is a ¹ H NMR chart of the compound (I) crystal form CS4

Figure 15 is a DSC chart of the compound (I) crystal form CS4

Figure 16 is a TGA diagram of the compound (I) crystal form CS4

Figure 17 is an XRPD pattern of the compound (I) crystal form CS5

Figure 18 is a ¹ H NMR chart of the compound (I) crystal form CS5

Figure 19 is a DSC chart of the compound (I) crystal form CS5

Figure 20 is a TGA diagram of the compound (I) crystal form CS5

Figure 21 is an XRPD pattern of the compound (I) crystal form CS6

Figure 22 is a ¹ H NMR chart of the compound (I) crystal form CS6

Figure 23 is a DSC chart of the compound (I) crystal form CS6

Figure 24 is a TGA diagram of the compound (I) crystal form CS6

Figure 25 is an XRPD overlay of the compound (I) crystal form CS1 before and after the polishing treatment (the upper graph is an XRPD pattern of the starting crystal form CS1, and the lower graph is an XRPD pattern of the crystal form CS1 after grinding)

Figure 26 is an XRPD overlay of the compound (I) crystal form CS3 before and after the grinding treatment (the upper graph is an XRPD pattern of the starting crystal form CS3, and the lower graph is an XRPD pattern of the crystal form CS3 after grinding)

Figure 27 is an XRPD overlay of the anhydrate A before and after the treatment of the compound (I) CN103328474A (the upper panel is an XRPD pattern of the anhydride A in the starting CN103328474A, and the lower panel is the XRPD of the anhydride A in the ground CN103328474A after grinding. Figure)

Figure 28 is an XRPD overlay of the compound (I) crystal form CS1 placed at 25 ° C / 60% RH for 3 weeks (the upper image shows the XRPD pattern of the crystalline form CS1 before placement, and the lower figure shows the crystal form CS1 after placement) XRPD diagram)

Figure 29 is an XRPD overlay of the compound (I) crystal form CS1 placed at 40 ° C / 75% RH for 3 weeks (the upper image shows the XRPD pattern of the crystalline form CS1 before placement, and the lower figure shows the crystal form CS1 after placement) XRPD diagram)

Figure 30 is an XRPD overlay of the compound (I) crystal form CS1 placed at 60 ° C / 75% RH for 3 weeks (the upper graph is the XRPD pattern of the crystalline form CS1 before placement, and the lower figure is the deposited form CS1 XRPD diagram)

Figure 31 is an XRPD overlay of the compound (I) crystal form CS3 placed at 25 ° C / 60% RH for 2 weeks (the upper image shows the XRPD pattern of the crystalline form CS3 before placement, and the lower figure shows the crystal form CS3 after placement) XRPD diagram)

Figure 32 is an XRPD overlay of Compound (I) Form CS3 placed at 40 °C / 75% RH for 2 weeks (the above figure is the XRPD pattern of the crystalline form CS3 before placement, and the lower figure is the placed form of CS3). XRPD diagram)

Fig. 33 is an XRPD stack of the compound (I) crystal form CS3 placed at 60 ° C / 75% RH for 2 weeks (the upper graph is the XRPD pattern of the crystal form CS3 before placement, and the lower graph is the crystal form CS3 after the placement) XRPD diagram)

Figure 34 is a DVS diagram of the compound (I) crystal form CS3

Figure 35 is a DVS diagram of an anhydride B in the compound (I) CN103328474A

Figure 36 is a PSD diagram of the compound (I) crystal form CS3

Figure 37 is a PSD diagram of anhydrate B in compound (I) CN103328474A

detailed description

The present invention will be further described in detail below with reference to specific embodiments, but the invention is not limited to the following examples. Conditions not specified in the examples are conventional conditions.

In the following examples, the test methods described are generally carried out under conventional conditions or conditions recommended by the manufacturer.

The terms used in the present invention are explained as follows:

XRPD: X-ray powder diffraction

DSC: Differential Scanning Calorimetry

TGA: Thermogravimetric Analysis

¹ H NMR: liquid NMR

The X-ray powder diffraction pattern of the present invention was collected on a Panalytical Empyrean X-ray powder diffractometer. The method parameters of the X-ray powder diffraction described in the present invention are as follows:

X-ray reflection parameters: Cu, Kα

Kα1

1.540598; Kα2

1.544426

Kα2/Kα1 intensity ratio: 0.50

Voltage: 45 volts (kV)

Current: 40 milliamps (mA)

Scan range: from 3.0 to 40.0 degrees

The differential scanning calorimetry (DSC) map of the present invention was acquired on a TA Q2000. The method parameters of the differential scanning calorimetry (DSC) described in the present invention are as follows:

Scan rate: 10 ° C / min

Protective gas: nitrogen

The thermogravimetric analysis (TGA) map of the present invention was taken on a TA Q5000. The method parameters of the thermogravimetric analysis (TGA) described in the present invention are as follows:

Scan rate: 10 ° C / min

Protective gas: nitrogen

H NMR data ⁽¹ HNMR) collected from a Bruker Avance II DMX 400M HZ NMR spectrometer. A sample of 1-5 mg was weighed and dissolved in 0.5 mL of deuterated dimethyl sulfoxide to prepare a solution of 2-10 mg/mL.

Example 1: Preparation of a solution of compound (I) in tetrahydrofuran

A compound of formula II ABT28 (5.0 g, 1.0 eq.), a compound of formula III ABT09 (3.6 g, 1.3 eq.), carbodiimide (2.2 g, 1.3 eq.) and dimethylaminopyridine (1.07 g, 1.0 eq.) were added. The mixture was stirred at room temperature for 5 minutes in 150 mL of dichloromethane, and then triethylamine (1.8 g, 2.0 eq.) was added, and the mixture was stirred at room temperature for 12 hours to give Compound (I). The reaction solution was washed with water several times, and the washed compound (I) dichloromethane solution was evaporated over tetrahydrofuran to obtain a tetrahydrofuran solution of the compound (I).

Example 2: Preparation method of crystal form CS1

The tetrahydrofuran solution of the compound (I) obtained in Example 1 was subjected to a sleeve distillation with an isopropyl acetate solvent to obtain a corresponding clear solution. The target clear solution was stirred and spontaneously devitrified at room temperature. After stirring for 12 hours, it was filtered, and the obtained solid was dried in a forced air oven at 50 ° C for 12 hours, and then the sample was subjected to a crystal form test.

Upon examination, the obtained crystalline solid is the crystalline form CS1 of the present invention, and the X-ray powder diffraction data thereof are shown in Fig. 1 and Table 1. The nuclear magnetic resonance spectrum is shown in Fig. 2, and the data is as follows: ¹ H NMR (400 MHz, DMSO) δ 11.71 (s, 1H), 8.62 (t, J = 5.9 Hz, 1H), 8.56 (d, J = 2.2 Hz, 1H), 8.04 (d, J = 2.6 Hz, 1H), 7.80 (dd, J = 9.2, 2.0 Hz, 1H), 7.58 - 7.42 (m, 3H), 7.34 (d, J = 8.4 Hz, 2H), 7.11 (d, J = 9.4 Hz, 1H), 7.04 (d, J = 8.4 Hz, 2H), 6.68 (dd, J = 9.0, 2.0 Hz, 1H), 6.39 (dd, J = 3.3, 1.9 Hz, 1H), 6.19 (d, J = 1.9 Hz, 1H), 3.85 (dd, J = 11.4, 2.9 Hz, 2H), 3.33 - 3.22 (m, 4H), 3.07 (s, 4H), 2.74 (d , J = 9.5 Hz, 2H), 2.17 (d, J = 23.0 Hz, 6H), 1.96 (d, J = 5.2 Hz, 2H), 1.61 (d, J = 11.8 Hz, 2H), 1.38 (t, J =6.3 Hz, 2H), 1.32–1.19 (m, 4H), 0.92 (s, 6H).

As shown in Fig. 3, the DSC of this crystal form showed an endothermic peak near 128 ° C, which is the dehydration of the crystal form CS1 near this temperature.

The TGA of this crystal form, as shown in Fig. 4, had a mass loss gradient of about 4.6% when heated to about 150 °C.

Table 1

Diffraction angle 2θ	d value	strength%
4.79	18.46	100.00
6.43	13.74	24.75
8.07	10.96	12.13
8.74	10.12	54.44
9.66	9.15	22.63
10.48	8.44	14.34
11.18	7.92	28.69
11.70	7.56	6.12
12.79	6.92	7.28
13.26	6.68	11.92
13.91	6.37	27.09
14.92	5.94	20.92
15.38	5.76	15.57
16.06	5.52	18.08
17.04	5.20	70.64
17.90	4.95	27.00
19.00	4.67	89.97
19.64	4.52	84.32
20.26	4.38	22.30
21.23	4.19	21.23
22.79	3.90	21.66
23.84	3.73	12.85
24.46	3.64	16.25
26.07	3.42	14.89
27.86	3.20	17.53
29.66	3.01	6.67
32.05	2.79	3.47

Example 3: Preparation method of crystal form CS2

The tetrahydrofuran solution of the compound (I) obtained in Example 1 was subjected to a sleeve distillation with a 1,4-dioxane solvent to obtain a corresponding clear solution. The target clear solution was spontaneously decanted for 12 hours at room temperature. After filtration, the obtained solid was subjected to a crystal form test after drying in a forced air oven at 50 ° C for 12 hours. Upon examination, the obtained crystalline solid is the crystalline form CS2 of the present invention, and its X-ray powder diffraction data is shown in FIG. 5 and Table 2. The nuclear magnetic resonance spectrum is shown in Fig. 6. The data are as follows: ¹ H NMR (400 MHz, DMSO) δ 11.68 (s, 1H), 8.60 (s, 1H), 8.55 (s, 1H), 8.04 (d, J = 2.6 Hz, 1H), 7.79 (d, J = 9.6 Hz, 1H), 7.50 (t, J = 9.8 Hz, 3H), 7.34 (d, J = 8.4 Hz, 2H), 7.10 (d, J = 9.2 Hz, 1H), 7.04 (d, J = 8.4 Hz, 2H), 6.68 (d, J = 8.9 Hz, 1H), 6.39 (s, 1H), 6.19 (s, 1H), 3.85 (d, J = 8.6 Hz, 2H), 3.57 (s, 4H), 3.31 - 3.22 (m, 4H), 3.07 (s, 4H), 2.74 (s, 2H), 2.16 (d, J = 17.5 Hz, 6H), 1.95 ( s, 2H), 1.61 (d, J = 10.9 Hz, 2H), 1.38 (t, J = 6.4 Hz, 2H), 1.23 (s, 4H), 0.92 (s, 6H).

The DSC of this crystal form is shown in Fig. 7. There is an endothermic peak near 144 ° C, and the endothermic peak is the solvent removal of the 1,4-dioxane solvate near this temperature.

The TGA of this crystal form, as shown in Fig. 8, has a mass loss gradient of about 4.4% when heated to about 160 °C. From the TGA calculation, about 0.5 mole of 1,4-dioxane is contained per mole of the crystalline form CS2.

Table 2

Diffraction angle 2θ	d value	strength%
5.43	16.28	100.00
6.91	12.79	1.32
7.99	11.06	27.61
9.84	8.99	3.20
10.88	8.13	2.89
11.98	7.39	1.93
13.37	6.62	2.23
14.45	6.13	14.43
15.07	5.88	6.00
15.50	5.72	2.45
15.90	5.57	11.63
16.28	5.44	8.28
16.93	5.24	6.32
17.40	5.10	4.18
18.12	4.90	12.28
18.69	4.75	21.03
19.57	4.54	20.67
20.04	4.43	14.91
20.82	4.27	5.57
21.58	4.12	8.13
21.95	4.05	3.41
22.80	3.90	5.65
23.72	3.75	3.81
24.89	3.58	6.93
25.88	3.44	5.14
26.44	3.37	7.68
27.58	3.23	3.16
28.32	3.15	2.65
29.33	3.04	1.82

30.34	2.95	2.97
32.12	2.79	0.83
33.20	2.70	1.14
34.37	2.61	0.73
36.96	2.43	0.80

Example 4: Preparation method of CS3

The tetrahydrofuran solution of the compound (I) prepared in Example 1 was spontaneously crystallized at room temperature for 3 hours. Then, a filter cake was obtained by filtration, the filter cake was added to a solvent of tetrahydrofuran, dissolved by heating, and then the solution was concentrated to 350 mL, and spontaneously crystallized for 3 hours at room temperature, finally filtered, and the obtained cake was dried at 50 °C. The crystal form of the dry product is tested. Upon examination, the obtained crystalline solid is the crystalline form CS3 of the present invention, and its X-ray powder diffraction data is shown in FIG. 9 and Table 3. The NMR spectrum is shown in Figure 10, and the data is as follows: ¹ H NMR (400 MHz, DMSO) δ 11.69 (s, 1H), 8.60 (s, 1H), 8.55 (s, 1H), 8.03 (d, J=2.4 Hz, 1H), 7.79 (d, J=8.2 Hz, 1H), 7.49 (d, J=8.8 Hz, 3H), 7.34 (d, J=8.4 Hz, 2H), 7.09 (s, 1H) , 7.04 (d, J = 8.4 Hz, 2H), 6.68 (d, J = 8.9 Hz, 1H), 6.39 (s, 1H), 6.19 (s, 1H), 3.85 (d, J = 10.8 Hz, 2H) , 3.31–3.23 (m, 4H), 3.07 (s, 4H), 2.74 (s, 2H), 2.16 (d, J = 15.6 Hz, 6H), 1.95 (s, 2H), 1.61 (d, J = 12.2) Hz, 2H), 1.38 (t, J = 6.5 Hz, 2H), 1.23 (s, 4H), 0.92 (s, 6H).

The DSC of this crystal form is as shown in Fig. 11, and the endothermic peak at around 139 ° C is the melting endothermic peak of the crystal form.

The TGA of this crystal form, as shown in Fig. 12, had a mass loss gradient of about 0.6% when heated to about 150 °C.

table 3

Diffraction angle 2θ	d value	strength%
5.20	16.98	100.00
7.73	11.44	0.95
9.12	9.70	1.69
10.38	8.53	80.44
11.36	7.79	5.81
12.34	7.17	3.32
13.78	6.42	1.39
14.27	6.21	2.09
15.21	5.82	16.34
15.62	5.67	8.98
16.40	5.40	4.74
17.24	5.14	2.36
17.94	4.94	4.90
19.26	4.61	12.21
19.66	4.52	30.56
20.93	4.24	10.09
21.74	4.09	21.89
22.85	3.89	3.76

23.80	3.74	3.72
24.25	3.67	18.64
24.54	3.63	13.64
25.22	3.53	6.37
26.17	3.41	12.71
29.37	3.04	13.46
30.96	2.89	1.85
34.67	2.59	4.28
36.98	2.43	1.17

Example 5: Preparation method of crystalline form CS4

100 mg of the compound (I) sample was weighed into a 3.0 mL glass vial, then 2 mL of methyl isobutyl ketone solvent was added, and stirred at room temperature for 24 hours, then filtered, and the wet product was vacuum dried at 35 ° C for 24 hours. Get dry goods. Upon examination, the obtained solid was the crystalline form CS4 of the present invention, and its X-ray powder diffraction data is shown in FIG. 13 and Table 4. The NMR spectrum is shown in Figure 14. The data is as follows: ¹ H NMR (400 MHz, DMSO) δ 11.69 (s, 1H), 8.61 (s, 1H), 8.56 (s, 1H), 8.04 (d, J = 2.6 Hz, 1H), 7.80 (d, J = 7.6 Hz, 1H), 7.50 (t, J = 10.8 Hz, 3H), 7.34 (d, J = 8.4 Hz, 2H), 7.11 (d, J = 9.1 Hz, 1H), 7.04 (d, J = 8.4 Hz, 2H), 6.68 (d, J = 10.9 Hz, 1H), 6.39 (s, 1H), 6.19 (s, 1H), 3.84 (d, J = 11.3 Hz, 2H), 3.25 (d, J = 11.3 Hz, 4H), 3.07 (s, 4H), 2.74 (s, 2H), 2.29 (d, J = 6.9 Hz, 1H), 2.16 (d, J = 16.6 Hz, 6H), 2.07–1.98 (m, 1H), 1.95 (s, 2H), 1.61 (d, J = 12.1 Hz, 3H), 1.38 (t, J = 6.4 Hz, 2H), 1.27 (dd, J=13.8, 9.7 Hz, 3H), 0.92 (s, 6H), 0.85 (d, J = 6.6 Hz, 2H).

The DSC of this crystal form, as shown in Fig. 15, showed an endothermic peak near 125 ° C, which is the solvent removal of the methyl isobutyl ketone solvate near this temperature.

The TGA of this crystal form is as shown in Fig. 16, and when it is heated to about 150 ° C, it has a mass loss of about 3.9%.

Table 4

Diffraction angle 2θ	d value	strength%
3.32	26.62	3.61
5.72	15.45	100.00
6.63	13.34	4.54
7.67	11.52	9.91
9.87	8.96	6.17
11.63	7.61	12.09
13.28	6.67	14.54
15.19	5.83	9.10
17.00	5.21	10.05
18.08	4.91	23.34
23.04	3.86	4.42

Example 6: Preparation method of crystalline form CS5

1.0 g of the compound (I) sample was dissolved in 20 mL of methyl isobutyl ketone solvent, stirred at 50 ° C for 18 hours, suction filtered, and the wet product was vacuum dried at 25 ° C for 24 hours to obtain a dry product. Upon examination, the obtained solid was the crystalline form CS5 of the present invention, and its X-ray powder diffraction data is shown in FIG. 17 and Table 5. The NMR spectrum is shown in Figure 18, and the data is as follows: ¹ H NMR (400 MHz, DMSO) δ 11.69 (s, 1H), 8.61 (s, 1H), 8.55 (d, J = 2.1 Hz, 1H) , 8.04 (d, J = 2.5 Hz, 1H), 7.80 (d, J = 9.0 Hz, 1H), 7.51 (dd, J = 12.8, 8.2 Hz, 3H), 7.34 (d, J = 8.4 Hz, 2H) , 7.11 (d, J = 9.1 Hz, 1H), 7.04 (d, J = 8.4 Hz, 2H), 6.68 (d, J = 9.1 Hz, 1H), 6.39 (dd, J = 3.3, 1.9 Hz, 1H) , 6.19 (s, 1H), 3.85 (d, J = 10.8 Hz, 2H), 3.25 (d, J = 11.6 Hz, 4H), 3.07 (s, 4H), 2.75 (s, 2H), 2.29 (d, J=6.9 Hz, 3H), 2.17 (d, J = 18.7 Hz, 6H), 2.06 (s, 3H), 2.01 (dd, J = 13.5, 6.7 Hz, 1H), 1.96 (d, J = 6.5 Hz, 2H), 1.61 (d, J = 12.8 Hz, 2H), 1.38 (t, J = 6.2 Hz, 2H), 1.27 (dd, J = 13.6, 10.0 Hz, 3H), 0.92 (s, 6H), 0.85 ( d, J = 6.7 Hz, 6H).

The DSC of this crystal form shows an endothermic peak near 140 ° C as shown in Fig. 19, which is the solvent removal of the methyl isobutyl ketone solvate near this temperature.

The TGA of this crystal form, as shown in Fig. 20, had a mass loss gradient of about 10.5% when heated to about 150 °C. Calculated from TGA, about 1.0 mole of methyl isobutyl ketone per mole of CS5.

table 5

Diffraction angle 2θ	d value	strength%
4.50	19.64	100.00
6.72	13.15	4.37
8.88	9.96	14.62
9.12	9.70	14.39
10.16	8.70	9.54
10.81	8.18	8.75
12.08	7.33	6.64
12.51	7.08	8.47
13.68	6.47	30.45
14.46	6.13	7.74
17.17	5.17	23.84
17.45	5.08	37.06
18.36	4.83	14.02
19.21	4.62	16.55
19.73	4.50	13.76
20.69	4.29	8.96
21.27	4.18	10.46
22.14	4.01	9.34
22.55	3.94	9.04
22.94	3.88	21.94
24.21	3.68	4.56
25.22	3.53	3.71
25.54	3.49	6.73
27.06	3.29	21.64

27.60	3.23	4.09
31.33	2.86	4.94
35.75	2.51	1.78
39.25	2.30	2.14

Example 7: Preparation method of crystalline form CS6

200 mg of the compound (I) sample was dissolved in 6.0 mL of methyl isobutyl ketone solvent, stirred at 80 ° C for 3 hours, and then suction filtered. After the filtrate was placed at 0 ° C for 2 hours, solids were precipitated and the solid was filtered. After drying at 25 ° C for 12 hours under vacuum, a dry product was obtained. Upon examination, the obtained solid was the crystalline form CS6 of the present invention, and its X-ray powder diffraction data is shown in FIG. 21 and Table 6. The nuclear magnetic resonance spectrum is shown in Fig. 22, and the data is as follows: ¹ H NMR (400 MHz, DMSO) δ 11.70 (s, 1H), 8.59 (d, J = 19.2 Hz, 1H), 8.57 (s, 1H) , 8.04 (d, J = 2.6 Hz, 1H), 7.80 (d, J = 9.0 Hz, 1H), 7.58 - 7.44 (m, 3H), 7.35 (d, J = 8.4 Hz, 2H), 7.15 - 7.06 ( m, 1H), 7.06 (s, 2H), 6.74 - 6.62 (m, 1H), 6.39 (dd, J = 3.3, 1.8 Hz, 1H), 6.19 (d, J = 2.0 Hz, 1H), 3.85 (dd , J=11.0, 2.9 Hz, 2H), 3.28 (dd, J = 18.2, 8.5 Hz, 4H), 3.08 (s, 4H), 2.76 (s, 2H), 2.30 (d, J = 6.9 Hz, 2H) , 2.17 (d, J = 20.7 Hz, 6H), 2.06 (s, 3H), 2.04 - 1.98 (m, 1H), 1.96 (s, 2H), 1.62 (d, J = 12.7 Hz, 2H), 1.39 ( t, J = 6.3 Hz, 2H), 1.29 - 1.21 (m, 4H), 0.93 (s, 6H), 0.85 (d, J = 6.7 Hz, 6H).

The DSC of this crystal form, as shown in Figure 23, has two endothermic peaks, and an endothermic peak begins to appear near 114 ° C. This endothermic peak is the solvent removal of the methyl isobutyl ketone solvate near this temperature.

The TGA of this crystal form, as shown in Fig. 24, had a mass loss gradient of about 10.4% when heated to about 150 °C. Calculated from TGA, about 1.0 mole of methyl isobutyl ketone per mole of CS6.

Table 6

Diffraction angle 2θ	d value	strength%
5.64	15.66	85.35
6.04	14.64	100.00
9.30	9.51	1.91
11.11	7.96	6.89
12.04	7.35	8.66
13.59	6.51	4.11
14.34	6.18	2.80
18.25	4.86	11.68
22.93	3.88	4.54

Example 8: Study on Mechanical Stability of Crystal Forms CS1 and CS3

Forms CS1 and CS3 and CN103328474A without crystal type A were placed in a mortar and manually ground for 5 minutes. The XRPD pattern of the crystal form before and after the test was tested. The test results are shown in Fig. 25 (XRPD stack before and after crystal form CS1), 26 (XRPD stack before and after crystal form CS3), and 27 (CN103328474A without crystal type A) XRPD overlay before and after).

It can be seen that the crystalline forms CS1 and CS3 of this patent have not changed after grinding, and the CN103328474A anhydrate A has been tested to be amorphous after grinding. It can be seen that the crystal forms CS1 and CS3 of the present invention have better mechanical grinding resistance than the anhydrate A in CN103328474A, so that they can maintain good stability and reliability in the post-treatment process such as tableting. Sex.

Better mechanical stability can be achieved under certain mechanical stresses while still maintaining stable physicochemical properties. The crystalline drug with better mechanical stability has low requirements on the crystallization equipment, requires no special post-treatment conditions, is more stable in the preparation process, can significantly reduce the development cost of the drug, enhance the quality of the drug, and has strong economic value.

Example 9: Physical and chemical stability of crystalline forms CS1 and CS3

The CS1 prepared by the present invention was placed under the conditions of 25 ° C / 60% RH, 40 ° C / 75% RH and 60 ° C / 75% RH for 3 weeks, and the stability of CS1 was examined. The test results are shown in Table 7, XRPD characterization. As shown in Figures 28, 29, and 30.

Table 7

The CS3 prepared by the present invention was placed under the conditions of 25 ° C / 60% RH, 40 ° C / 75% RH and 60 ° C / 75% RH for 2 weeks, and the stability of CS3 was examined. The test results are shown in Table 8, XRPD characterization. As shown in Figures 31, 32, and 33.

Table 8

The test results show that the crystalline form CS1 and the crystalline form CS3 of the invention have good physical and chemical stability, and good physical and chemical stability can ensure that the chemical degradation of the drug is maintained at a low level, and the raw material drug itself and the raw material drug are in the preparation. Stability and quality indicators. Better physical stability can reduce the risk of drug dissolution rate and bio-profit change due to crystal form changes, which is of great significance to ensure drug efficacy and safety and prevent adverse drug reactions.

Example 10: Dynamic Solubility Study of Forms CS1 and CS3

The crystalline forms CS1, CS3 and CN103328474A anhydrate B of the present invention were prepared into a saturated solution by using FaSSIF, FeSSIF and water, respectively, and the samples in the saturated solution were respectively tested by high performance liquid chromatography (HPLC) after 1 hour, 4 hours and 24 hours. The content (μg/mL) is shown in Table 9.

Table 9

ND: below detection limit

From the results of the dynamic solubility of Table 9, it is known that the crystalline forms CS1, CS3 and CN103328474A anhydrate B of the present invention have better solubility, and the crystal forms CS1 and CS3 of the present invention are in FaSSIF, FeSSIF and H at 1 hour, 4 hours, and 24 hours. _The solubility of ₂ O in three biological media was significantly higher than that of CN103328474A anhydrate B, which was increased by 1.5-5.5 times. The significant increase in solubility can effectively improve the bioavailability of the drug, lower dosage, thereby reducing the side effects of the drug and improving the safety of the drug, and providing safety protection while giving people treatment.

Example 11: Study on the wettability of crystalline form CS3

About 25 mg of each of the crystalline form CS3 and CN103328474A anhydrate B of the present invention was subjected to dynamic moisture adsorption (DVS) to test its wettability at 25 °C. The experimental results are shown in Table 10. The wettability DVS patterns of Forms CS3 and CN103328474A Anhydrate B are shown in Figures 34 and 35.

Table 10

With reference to the definition of wettability in the Chinese Pharmacopoeia 2015 General Regulation 9103, CN103328474A anhydrate B belongs to the hygroscopicity, and the crystalline form CS3 of the present invention is slightly wetted, and it can be seen that the crystalline form CS3 is compared with the CN103328474A anhydrate B. It has lower moisture absorption and is suitable for later product development and storage.

Defining the characteristics of wettability and the definition of wetting weight gain (Chinese Pharmacopoeia 2015 General Principles 9103 Guidelines for Drug Wetness Test, Experimental Conditions: 25 °C ± 1 °C, 80% Relative Humidity):

Deliquescence: absorb enough water to form a liquid

Very hygroscopic: the wetting weight gain is not less than 15%

Humidity: Wet weight gain is less than 15% but not less than 2%

Slightly wettability: wetting gain is less than 2% but not less than 0.2%

No or almost no wettability: wetting gain is less than 0.2%

Example 12: Study on particle properties of crystalline form CS3

PSD test

The PSD test was performed on the crystalline form CS3 of the present invention and CN10328474A anhydrate B, respectively. The PSD data of the crystalline form CS3 and the CN103328474A anhydrous B of the present invention are shown in Table 11, and the PSD diagram is shown in Figs.

Table 11

Mv: represents the average particle size by volume

SD: indicates standard deviation

D10: indicates that the particle size distribution (volume distribution) accounts for 10% of the particle size of the anhydrate B

D50: indicates the particle diameter corresponding to the particle size distribution (volume distribution), which is also called the median diameter.

D90: indicates the particle size distribution (volume distribution) accounts for 90% of the particle size

As a result of the above test, the crystal form CS3 of the present invention has an average particle diameter of about 6 μm and is normally distributed, and has a uniform particle dispersion property and a narrow particle size distribution. However, CN103328474A anhydrate B has different particle sizes, large differences, no normal distribution, and poor particle uniformity.

A narrower particle size distribution improves the uniformity of the drug substance components in the formulation, while making the difference between different batches of the formulation smaller, such as more uniform dissolution; smaller crystal size can increase the specific surface area of the drug, and increase The dissolution rate of the drug is beneficial to the absorption of the drug, thereby improving the bioavailability. Large clusters of crystals are often susceptible to entrapment of residual solvents or other impurities. Moreover, in the preparation of the preparation, the bulk crystal powder cannot be uniformly dispersed, and it is difficult to mix uniformly with the auxiliary material, which is disadvantageous for the preparation of the preparation.

The above embodiments are merely illustrative of the technical concept and the features of the present invention, and the purpose of the present invention is to enable those skilled in the art to understand the present invention and to implement the present invention, and the scope of the present invention is not limited thereto. Equivalent variations or modifications made in accordance with the spirit of the invention are intended to be included within the scope of the invention.

Claims (16)

1. a crystal form CS1 of the compound (I), characterized in that

   

   Its X-ray powder diffraction pattern (CuKα radiation) has a characteristic peak at a 2θ value of 4.8°±0.2°, 17.0°±0.2°, and 19.0°±0.2°.
2. The crystal form CS1 according to claim 1, further characterized in that the X-ray powder diffraction pattern is further one or more of a 2θ value of 8.7 ° ± 0.2 °, 13.9 ° ± 0.2 °, and 19.6 ° ± 0.2 °. There is a diffraction peak at the place.
3. The crystal form CS1 according to claim 1 or 2, wherein the X-ray powder diffraction pattern is further in a position where the 2θ value is 6.4 ° ± 0.2 °, 11.2 ° ± 0.2 °, 17.9 ° ± 0.2 °. Or multiple points with diffraction peaks.
4. A method for preparing a crystalline form CS1 according to claim 1, characterized in that the method comprises: subjecting a target solvent set to steam displacement using a tetrahydrofuran solution of the compound (I), and crystallization at 20-40 ° C to obtain The target solvent is a C ₃ -C ₈ ester solvent.
5. The process according to claim 4, wherein the ester solvent is ethyl formate, ethyl acetate, isopropyl acetate or a mixed solvent thereof.
6. The method according to claim 5, wherein the ester solvent is isopropyl acetate.
7. The preparation method according to claim 4, wherein the crystallization time is 6 to 24 hours.
8. The production method according to claim 7, wherein the crystallization time is 12 hours.
9. A crystal form CS3 of the compound (I), characterized in that the X-ray powder diffraction pattern (CuKα radiation) has a characteristic peak at a 2θ value of 10.4°±0.2°, 15.2°±0.2°, and 21.7°±0.2°. .
10. The crystal form CS3 according to claim 9, wherein the X-ray powder diffraction pattern has diffraction at one or more of 5.2 ° ± 0.2 °, 19.7 ° ± 0.2 °, and 29.4 ° ± 0.2 °. peak.
11. The crystal form CS3 according to claim 9 or 10, wherein the X-ray powder diffraction pattern is further in one or more of 20.9 ° ± 0.2 °, 24.3 ° ± 0.2 °, and 26.2 ° ± 0.2 °. Has a diffraction peak.
12. A method for preparing a crystalline form CS3 according to claim 9, wherein the method comprises: stirring and crystallization using a solution of the compound (I) in tetrahydrofuran at 20-40 ° C, and filtering the filter cake again. The tetrahydrofuran solvent is dissolved by heating, then concentrated, and obtained by crystallization at 20 to 40 ° C.
13. The method according to claim 12, wherein the crystallization time is 3 to 24 hours.
14. The preparation method according to claim 13, wherein the crystallization time is 3 hours.
15. A pharmaceutical composition comprising a therapeutically effective amount of Form CS1 of Claim 1 or Form CS3 of Claim 9 or a mixture thereof and a pharmaceutically acceptable adjuvant.
16. Use of the crystalline form CS3 of claim 1 or the crystalline form CS3 of claim 9 or a mixture thereof or the pharmaceutical composition of claim 15 for the manufacture of a medicament for the treatment of chronic lymphocytic leukemia.

Images