Patent Application
20240043416
The present application is based on and claims the right of priority for the application with the application no. being CN 202110157185.7 and the filing date being 4 Feb. 2021, and the disclosure of the present application is incorporated herein by reference in its entirety.
The present application relates to a pharmaceutically acceptable salt of an HA inhibitor compound (1S,2S)-2-fluoro-N-(2-(2-(4-((R)-(5-methyl-2H-tetrazol-2-yl)(phenyl)methyl)piperidine-1-carbonyl)pyridin-4-yl)benzo[d]oxazol-5-y1)cyclopropane-1-carboxamide, or a hydrate or solvate, a crystal form thereof, a preparation method therefor, a pharmaceutical composition thereof and the use thereof in the preparation of a hemagglutinin (HA) inhibitor.
Influenza (flu for short) is an acute respiratory infection caused by influenza viruses and is also a highly infectious and fast-transmitting disease. At present, anti-influenza virus drugs mainly target the hemagglutinin receptor, neuraminidase and matrix protein of the viral envelope.
According to the surface antigen hemagglutinin (HA) and neuraminidase (NA) protein structures and gene characteristics, influenza A viruses are divided into different subtypes. Hemagglutinins that have been discovered currently comprise 18 subtypes (H1 to H18), and neuraminidases that have been discovered currently comprise 10 subtypes (N1 to N10). The entry of a virus into a host cell is the first important step of the initiation of a viral replication cycle. Influenza virus protein hemagglutinins can recognize the potential binding site of sialic acid (SA) (N-acetylneuraminic acid) on glycoproteins of a host cell. The HAs contained in influenza viruses which infect humans have high specificity to α2-6SA. After HAs bind to receptors, the viruses are endocytosed; and the acidic pH of endosomes causes changes on the conformation of HA proteins, thereby regulating the internal fusion of the viruses and recipient cells and releasing RNPs of the viruses into cytoplasm. Therefore, by using HAs as targets and binding to the HAs, the conformational change of HA2 caused by low pH conditions is inhibited, thereby inhibiting the process of fusion of viral envelopes with host endosomal membranes, which has become a new anti-influenza virus strategy. Currently, there are multiple vaccines for HAs in the clinical stage, such as CR9114 (WO 2013/007770) and CR6261 (WO 2008/028946). Small molecule compounds with different structural features for the treatment of influenza have also been reported in the literature. A series of benzisoxazole compounds for the treatment of influenza are reported in WO 2012/144752.
The present application provides a salt having the following structure (labelled as compound A), or a hydrate, a solvate or a crystal form of a salt thereof,
The salt of compound A, and the hydrate, solvate or crystal form of the salt thereof have better solubility and stability than a free base compound, can be very stable in a diluent (solvent) and are capable of resisting high temperature, high humidity and strong light during the preparation of a medicament or a composition thereof (which are suitable for the preparation of pharmaceutical dosage forms), and also have better pharmacokinetics and bioavailability than a free base compound.
In particular, the present application provides a salt of a compound represented by formula (I), or a hydrate or solvate of a salt thereof:
The salt is selected from hydrochloride, hydrobromide, 2-naphthalenesulfonate, benzenesulfonate, methanesulfonate, p-toluenesulfonate, hemi-1,5-naphthalene disulfonate, succinate, citrate or malate.
Further, the salt of compound A is selected from hydrochloride, hydrobromide, 2-naphthalenesulfonate or hemi-1,5-naphthalene disulfonate, preferably the crystal form thereof.
Furthermore, the salt of compound A is the hemi-1,5-naphthalene disulfonate.
The present application provides the salt of compound A or the hydrate or solvate of the salt thereof, wherein the salt has a structure selected from
The present application further provides a method for preparing the hemi-1,5-naphthalene disulfonate of compound A, comprising the steps of
The present application further provides a method for preparing the 2-naphthalenesulfonate of compound A, comprising the steps of
The present application further provides a method for preparing the hydrochloride of compound A, comprising the steps of
The present application further provides a method for preparing the hydrobromide of compound A, comprising the steps of
The present application further provides a method for preparing the benzenesulfonate of compound A, comprising the steps of
The present application further provides a method for preparing the p-toluenesulfonate of compound A, comprising the steps of
The present application further provides a method for preparing the p-methanesulfonate of compound A, comprising the steps of
The present application further provides a method for preparing the acetate of compound A, comprising the steps of
The present application further provides a method for preparing the fumarate of compound A, comprising the steps of
With the same method as the fumarate of compound A, the malonate, succinate, benzoate, citrate, malate and L-tartrate can be prepared by selecting the corresponding acids.
The method for preparing the salt of compound A of the present application may further involve adding an anti-solvent, wherein the anti-solvent can be selected from one of isopropyl ether, diethyl ether, water and n-heptane.
In the method for preparing the salt of compound A of the present application, the molar ratio of compound A to an acid is 1:5-1:1.1, further 1:2-1:1.2, and more further 1:1.5-1:1.2.
The method for preparing the salt of compound A of the present application is performed at normal temperature, further 15° C.-30° C.
The present application also relates to a hemi-1,5-naphthalene disulfonate of compound A, which is in the form of a crystal (crystal form I of hemi-1,5-naphthalene disulfonate) and has an X-ray powder diffraction pattern comprising characteristic diffraction peaks at 5.3°±0.2°, 13.4°±0.2°, 17.4°±0.2°, 18.5°±0.2°, 20.4°±0.2° and 23.6°±0.2° 2θ, as determined by using Cu-Kα radiation.
Further, the crystal form I of the hemi-1,5-naphthalene disulfonate of compound A provided by the present application has an X-ray powder diffraction pattern further comprising characteristic diffraction peaks at 7.0°±0.2°, 9.8°±0.2°, 10.6°±0.2°, 12.7°±0.2°, 14.8°±0.2°, 22.2°±0.2° and 23.1°±0.2° 2θ.
Furthermore, the crystal form I of the hemi-1,5-naphthalene disulfonate of compound A provided by the present application has an X-ray powder diffraction pattern further comprising characteristic diffraction peaks at 14.1°±0.2°, 16.0°±0.2° and 21.5°±0.2° 2θ.
Furthermore, the crystal form I of the hemi-1,5-naphthalene disulfonate of compound A provided by the present application has an X-ray powder diffraction pattern further comprising characteristic diffraction peaks at 9.8°±0.2°, 11.6°±0.2°, 16.6°±0.2°, 17.9°±0.2°, 19.2°±0.2° and 27.6°±0.2° 2θ.
Furthermore, the crystal form I of the hemi-1,5-naphthalene disulfonate of compound A provided by the present application has an X-ray powder diffraction pattern further comprising characteristic diffraction peaks at 18.9°±0.2°, 21.8°±0.2°, 24.9°±0.2°, 27.3°±0.2°, 27.9°±0.2°, 28.33°±0.2°, 29.0°±0.2° and 33.4°±0.2° 2θ.
Table 1 shows 2θ values and corresponding intensities in the X-ray powder diffraction pattern of the crystal form I of the hemi-1,5-naphthalene disulfonate of compound A provided by the present application, wherein the error range of 2θ is ±0.2°.
Further, the crystal form I of the hemi-1,5-naphthalene disulfonate of compound A provided by the present application has an X-ray powder diffraction pattern substantially as shown in
The crystal form I of the hemi-1,5-naphthalene disulfonate of compound A provided by the present application has a differential scanning calorimetry (DSC) pattern showing an endothermic curve, wherein Tstart=188.78° C., Tpeak=198.44° C. and ΔH=46.09 J/g.
The crystal form I of the hemi-1,5-naphthalene disulfonate of compound A provided by the present application has a differential scanning calorimetry (DSC) pattern showing a melting point of 188.78° C.
The crystal form I of the hemi-1,5-naphthalene disulfonate of compound A provided by the present application has a differential scanning calorimetry (DSC) pattern substantially as shown in
The crystal form I of the hemi-1,5-naphthalene disulfonate of compound A provided by the present application has a thermogravimetric analysis (TGA) curve showing a weight loss of 4.017% below 150° C. and showing a decomposition temperature of 213.86° C.
The crystal form I of the hemi-1,5-naphthalene disulfonate of compound A provided by the present application has a thermogravimetric analysis (TGA) curve substantially as shown in
The crystal form I of the hemi-1,5-naphthalene disulfonate of compound A provided by the present application is an anhydride.
The present application also provides a hydrochloride of compound A, which is in the form of a crystal (crystal form I of hydrochloride) and has an X-ray powder diffraction pattern comprising characteristic diffraction peaks at 5.9°±0.2°, 11.2°±0.2°, 11.7°±0.2°, 17.6°±0.2°, 18.2°±0.2°, 21.9°±0.2° and 26.8°±0.2° 2θ, as determined by using Cu-Kα radiation.
Further, the crystal form I of the hydrochloride of compound A provided by the present application has an X-ray powder diffraction pattern further comprising characteristic diffraction peaks at 7.1°±0.2°, 16.3°±0.2°, 18.6°±0.2°, 22.3°±0.2° and 23.8°±0.2° 2θ.
Furthermore, the crystal form I of the hydrochloride of compound A provided by the present application has an X-ray powder diffraction pattern further comprising characteristic diffraction peaks at 13.3°±0.2°, 14.2°±0.2°, 15.7°±0.2°, 20.3°±0.2°, 21.3°±0.2°, 24.8°±0.2°, 25.4°±0.2°, 27.2°±0.2° and 27.7°±0.2° 2θ.
Furthermore, the crystal form I of the hydrochloride of compound A provided by the present application has an X-ray powder diffraction pattern further comprising characteristic diffraction peaks at 6.6±0.2°, 8.1±0.2°, 10.3±0.2°, 12.9±0.2°, 15.9±0.2°, 21.3±0.2°, 23.0±0.2° and 23.6±0.2° 2θ.
Furthermore, the crystal form I of the hydrochloride of compound A provided by the present application has an X-ray powder diffraction pattern further comprising characteristic diffraction peaks at 21.0±0.2°, 24.3±0.2°, 24.8±0.2°, 26.4±0.2°, 26.1±0.2°, 27.2±0.2°, 28.8±0.2° and 29.8±0.2° 2θ.
Table 2 shows 2θ values and corresponding intensities in the X-ray powder diffraction pattern of the crystal form I of the hydrochloride of compound A provided by the present application, wherein the error range of 2θ is ±0.2°.
Further, the crystal form I of the hydrochloride of compound A provided by the present application has an X-ray powder diffraction pattern substantially as shown in
The crystal form I of the hydrochloride of compound A provided by the present application has a differential scanning calorimetry (DSC) pattern showing an endothermic curve, wherein Tstart=118.97° C., Tpeak=126.16° C. and ΔH=23.18 J/g.
Further, the crystal form I of the hydrochloride of compound A provided by the present application has a differential scanning calorimetry (DSC) pattern showing a melting point of 118.97° C.
Further, the crystal form I of the hydrochloride of compound A provided by the present application has a differential scanning calorimetry pattern substantially as shown in
The crystal form I of the hydrochloride of compound A provided by the present application has a thermogravimetric analysis (TGA) curve showing a weight loss of 3.202% below 150° C. and showing a decomposition temperature of 171.90° C.
Further, the crystal form I of the hydrochloride of compound A provided by the present application has a thermogravimetric analysis (TGA) curve substantially as shown in
The crystal form of the hydrochloride of compound A provided by the present application may exist in the form of a solvate.
The present application also provides a hydrobromide of compound A, which is in the form of a crystal (crystal form I of hydrobromide) and has an X-ray powder diffraction pattern comprising characteristic diffraction peaks at 6.0°±0.2°, 7.2°±0.2°, 9.0°±0.2°, 12.0°±0.2°, 14.8°±0.2° and 17.6°±0.2° 2θ, as determined by using Cu-Kα radiation.
Further, the crystal form I of the hydrobromide of compound A in the present application has an X-ray powder diffraction pattern further comprising characteristic diffraction peaks at 17.3°±0.2°, 18.0°±0.2°, 21.2°±0.2°, 21.5°±0.2°, 24.2°±0.2° and 26.5°±0.2° 2θ.
Further, the crystal form I of the hydrobromide of compound A in the present application has an X-ray powder diffraction pattern further comprising characteristic diffraction peaks at 16.9°±0.2°, 18.6°±0.2°, 19.0°±0.2°, 20.2°±0.2° and 28.0°±0.2° 2θ.
Further, the crystal form I of the hydrobromide of compound A in the present application has an X-ray powder diffraction pattern further comprising characteristic diffraction peaks at 14.4°±0.2°, 20.4°±0.2°, 22.1°±0.2°, 22.5°±0.2°, 23.7°±0.2°, 24.8°±0.2°, 27.1°±0.2° and 28.4°±0.2° 2θ.
Further, the crystal form I of the hydrobromide of compound A in the present application has an X-ray powder diffraction pattern further comprising characteristic diffraction peaks at 26.1°±0.2° and 27.1°±0.2° 2θ.
Table 3 shows 2θ values and corresponding intensities in the X-ray powder diffraction pattern of the crystal form I of the hydrobromide of compound A provided by the present application, wherein the error range of 2θ is ±0.2°.
Further, the crystal form I of the hydrobromide of compound A provided by the present application has an X-ray powder diffraction pattern substantially as shown in
The crystal form I of the hydrobromide of compound A provided by the present application has a differential scanning calorimetry (DSC) pattern showing an endothermic curve, wherein Tstart=179.68° C., Tpeak=187.40° C. and ΔH=8.189 J/g.
Further, the crystal form I of the hydrobromide of compound A provided by the present application has a differential scanning calorimetry (DSC) pattern showing a melting point of 179.68° C.
Further, the crystal form I of the hydrobromide of compound A provided by the present application has a differential scanning calorimetry (DSC) pattern substantially as shown in
The crystal form I of the hydrobromide of compound A provided by the present application has a thermogravimetric analysis (TGA) curve showing a weight loss of 3.584% below 100° C. and a weight loss of 7.033% between 100° C.-150° C. and showing a decomposition temperature of 185.29° C.
Further, the crystal form I of the hydrobromide of compound A provided by the present application has a thermogravimetric analysis (TGA) curve substantially as shown in
The present application also relates to a hydrobromide of compound A, which is in the form of a crystal (crystal form II of hydrobromide) and has an X-ray powder diffraction pattern comprising characteristic diffraction peaks at 6.5°±0.2°, 7.3°±0.2°, 12.2°±0.2°, 12.9°±0.2° and 16.0°±0.2° 2θ, as determined by using Cu-Kα radiation.
Further, the crystal form II of the hydrobromide of compound A in the present application has an X-ray powder diffraction pattern further comprising characteristic diffraction peaks at 6.1°±0.2°, 17.4°±0.2°, 18.3°±0.2°, 20.4°±0.2°, 22.4°±0.2°, 24.8°±0.2° and 28.2°±0.2° 2θ.
Further, the crystal form II of the hydrobromide of compound A in the present application has an X-ray powder diffraction pattern further comprising characteristic diffraction peaks at 13.8°±0.2°, 14.5°±0.2°, 15.2°±0.2°, 19.1°±0.2°, 19.9°±0.2°, 21.4°±0.2°, 21.8°±0.2°, 23.1°±0.2°, 23.6°±0.2°, 25.5°±0.2°, 26.0°±0.2° and 26.5°±0.2° 2θ.
Table 4 shows 2θ values and corresponding intensities in the X-ray powder diffraction pattern of the crystal form II of the hydrobromide of compound A provided by the present application, wherein the error range of 2θ is ±0.2°.
Further, the crystal form II of the hydrobromide of compound A provided by the present application has an X-ray powder diffraction pattern substantially as shown in
The crystal form of the hydrobromide of compound A provided by the present application may exist in the form of a solvate.
The crystal form I of the hydrobromide of compound A provided by the present application may exist in the form of a solvate.
The present application also provides a 2-naphthalenesulfonate of compound A, which is in the form of a crystal (crystal form I of 2-naphthalenesulfonate) and has an X-ray powder diffraction pattern comprising characteristic diffraction peaks at 4.7°±0.2°, 9.4°±0.2°, 17.2°±0.2°, 21.2°±0.2° and 23.4°±0.2° 2θ, as determined by using Cu-Kα radiation.
Further, the crystal form I of the 2-naphthalenesulfonate of compound A provided by the present application has an X-ray powder diffraction pattern further comprising characteristic diffraction peaks at 6.3°±0.2°, 6.8°±0.2°, 7.8°±0.2°, 13.4°±0.2°, 16.5°±0.2°, 19.2°±0.2° and 20.1°±0.2° 2θ.
Further, the crystal form I of the 2-naphthalenesulfonate of compound A provided by the present application has an X-ray powder diffraction pattern further comprising characteristic diffraction peaks at 14.9°±0.2°, 15.3°±0.2°, 15.7°±0.2°, 24.3°±0.2°, 25.1°±0.2° and 26.1°±0.2° 2θ.
Further, the crystal form I of the 2-naphthalenesulfonate of compound A provided by the present application has an X-ray powder diffraction pattern further comprising characteristic diffraction peaks at 11.9°±0.2°, 12.3°±0.2°, 12.5°±0.2°, 13.9°±0.2° and 17.5°±0.2° 2θ.
Further, the crystal form I of the 2-naphthalenesulfonate of compound A provided by the present application has an X-ray powder diffraction pattern further comprising characteristic diffraction peaks at 18.4°±0.2°, 19.7°±0.2°, 21.9°±0.2°, 23.8°±0.2°, 26.1°±0.2° and 27.3°±0.2° 2θ.
Table 5 shows 2θ values and corresponding intensities in the X-ray powder diffraction pattern of the crystal form I of the 2-naphthalenesulfonate of compound
A provided by the present application, wherein the error range of 2θ is ±0.2°.
Further, the crystal form I of the 2-naphthalenesulfonate of compound A provided by the present application has an X-ray powder diffraction pattern substantially as shown in
The crystal form I of the 2-naphthalenesulfonate of compound A provided by the present application has a differential scanning calorimetry (DSC) pattern showing an endothermic curve, wherein Tstart=143.43° C., Tpeak=152.36° C. and ΔH=46.11 J/g.
Further, the crystal form I of the 2-naphthalenesulfonate of compound A provided by the present application has a melting point of 143.43° C.
Further, the crystal form I of the 2-naphthalenesulfonate of compound A provided by the present application has a differential scanning calorimetry (DSC) pattern substantially as shown in
The crystal form I of the 2-naphthalenesulfonate of compound A provided by the present application has a thermogravimetric analysis (TGA) curve showing a weight loss of 12.17% below 150° C. and showing a decomposition temperature of 211.99° C.
The crystal form I of the 2-naphthalenesulfonate of compound A provided by the present application has a thermogravimetric analysis curve substantially as shown in
The crystal form I of the 2-naphthalenesulfonate of compound A provided by the present application is an anhydride.
The present application also relates to a 2-naphthalenesulfonate of compound A, which is in the form of a crystal (crystal form II of 2-naphthalenesulfonate) and has an X-ray powder diffraction pattern comprising characteristic diffraction peaks at 5.6°±0.2°, 11.2°±0.2°, 14.1°±0.2°, 16.0°±0.2°, 22.8°±0.2° and 26.8°±0.2° 2θ, as determined by using Cu-Kα radiation.
The crystal form II of the 2-naphthalenesulfonate of compound A of the present application has an X-ray powder diffraction pattern further comprising characteristic diffraction peaks at 4.5°±0.2°, 6.2°±0.2°, 6.8°±0.2°, 8.4°±0.2°, 10.4°±0.2°, 15.3°±0.2°, 15.6°±0.2°, 19.0°±0.2°, 19.6°±0.2° and 25.5±0.2° 2θ, as determined by using Cu-Kα radiation.
The crystal form II of the 2-naphthalenesulfonate of compound A of the present application has an X-ray powder diffraction pattern further comprising characteristic diffraction peaks at 12.4°±0.2°, 12.7°±0.2°, 16.7°±0.2°, 17.2°±0.2°, 17.5°±0.2°, 18.0°±0.2°, 20.8°±0.2°, 21.8°±0.2°, 23.5°±0.2° and 24.3°±0.2° 2θ, as determined by using Cu-Kα radiation.
Table 6 shows 2θ values and corresponding intensities in the X-ray powder diffraction pattern of the crystal form II of the 2-naphthalenesulfonate of compound A provided by the present application, wherein the error range of 2θ is ±0.2°.
The crystal form II of the 2-naphthalenesulfonate of compound A of the present application has an X-ray powder diffraction pattern substantially as shown in
The salt or crystal form of the present application accounts for approximately 5 wt % to approximately 100 wt % of a bulk drug.
In some embodiments, the salt or crystal form of the present application accounts for approximately 10 wt % to approximately 100 wt % of the bulk drug.
In some embodiments, the salt or crystal form of the present application accounts for approximately 15 wt % to approximately 100 wt % of the bulk drug.
In some embodiments, the salt or crystal form of the present application accounts for approximately 20 wt % to approximately 100 wt % of the bulk drug.
In some embodiments, the salt or crystal form of the present application accounts for approximately 25 wt % to approximately 100 wt % of the bulk drug.
In some embodiments, the salt or crystal form of the present application accounts for approximately 30 wt % to approximately 100 wt % of the bulk drug.
In some embodiments, the salt or crystal form of the present application accounts for approximately 35 wt % to approximately 100 wt % of the bulk drug.
In some embodiments, the salt or crystal form of the present application accounts for approximately 40 wt % to approximately 100 wt % of the bulk drug.
In some embodiments, the salt or crystal form of the present application accounts for approximately 45 wt % to approximately 100 wt % of the bulk drug.
In some embodiments, the salt or crystal form of the present application accounts for approximately 50 wt % to approximately 100 wt % of the bulk drug.
In some embodiments, the salt or crystal form of the present application accounts for approximately 55 wt % to approximately 100 wt % of the bulk drug.
In some embodiments, the salt or crystal form of the present application accounts for approximately 60 wt % to approximately 100 wt % of the bulk drug.
In some embodiments, the salt or crystal form of the present application accounts for approximately 65 wt % to approximately 100 wt % of the bulk drug.
In some embodiments, the salt or crystal form of the present application accounts for approximately 70 wt % to approximately 100 wt % of the bulk drug.
In some embodiments, the salt or crystal form of the present application accounts for approximately 75 wt % to approximately 100 wt % of the bulk drug.
In some embodiments, the salt or crystal form of the present application accounts for approximately 80 wt % to approximately 100 wt % of the bulk drug.
In some embodiments, the salt or crystal form of the present application accounts for approximately 85 wt % to approximately 100 wt % of the bulk drug.
In some embodiments, the salt or crystal form of the present application accounts for approximately 90 wt % to approximately 100 wt % of the bulk drug.
In some embodiments, the salt or crystal form of the present application accounts for approximately 95 wt % to approximately 100 wt % of the bulk drug.
In some embodiments, the salt or crystal form of the present application accounts for approximately 98 wt % to approximately 100 wt % of the bulk drug.
In some embodiments, the salt or crystal form of the present application accounts for approximately 99 wt % to approximately 100 wt % of the bulk drug.
In some embodiments, the salt or crystal form of the present application substantially accounts for 100 wt % of the bulk drug, that is, the bulk drug is substantially a pure phase salt or crystal.
It can be understood that, as is well known in the field of differential scanning calorimetry (DSC), a melting peak height of a DSC curve depends on many factors related to sample preparation and geometric shapes of instruments, and a peak position is relatively insensitive to experiment details. Therefore, in some embodiments, the crystallized compounds of the present application have DSC patterns comprising characteristic peak positions, wherein the DSC patterns have substantially the same properties as those provided in the drawings of the present application, with an error tolerance of ±3° C.
The present application also relates to a pharmaceutical composition comprising a therapeutically effective amount of the salt of compound A, or the hydrate, solvate or crystal form of the salt thereof according to the present application, and a pharmaceutically acceptable carrier or excipient.
The present application also relates to the use of the salt of compound A, or the hydrate, solvate or crystal form of the salt thereof, or the pharmaceutical composition, in the preparation of a medicament for preventing and/or treating influenza.
The present application also relates to a method for treating and/or preventing influenza, comprising administering to a subject in need thereof a therapeutically and/or prophylactically effective amount of the salt of compound A, or the hydrate, solvate or crystal form of the salt thereof, or the pharmaceutical composition according to the present application.
The present application also relates to the salt of compound A, or the hydrate, solvate or crystal form of the salt thereof, or the pharmaceutical composition, for use in the treatment and/or prevention of influenza.
The present application further provides a composition for treating and/or preventing influenza, comprising the salt of compound A, or the hydrate, solvate or crystal form of the salt thereof, or the pharmaceutical composition.
Unless stated to the contrary, the terms used in the description and claims have the following meanings.
The carbon, hydrogen, oxygen, sulphur, nitrogen and halogen involved in the groups and compounds of the present application all comprise isotopes thereof, and are optionally further substituted with one or more of the corresponding isotopes thereof, wherein the isotopes of carbon comprise 12C, 13C and 1j; the isotopes of hydrogen comprise protium (H), deuterium (D, also known as heavy hydrogen), and tritium (T, also known as superheavy hydrogen); the isotopes of oxygen comprise 16O, 17O and 18O; the isotopes of sulphur comprise 32S, 33S, 34S and 36S; the isotopes of nitrogen comprise 14N and 15N; the isotope of fluorine comprises 19F; the isotopes of chlorine comprise 35Cl and 37Cl; and the isotopes of bromine comprise 79Br and 81Br.
The “effective amount” means an amount of a compound that causes a physiological or medical response in a tissue, system or subject and is a desirable amount, including the amount of a compound that is, when administered to a subject to be treated, sufficient to prevent occurrence of one or more symptoms of the disease or condition to be treated or to reduce the symptom(s) to a certain degree.
The “IC50” refers to the half maximal inhibitory concentration, i.e., a concentration where half of the maximum inhibitory effect is achieved.
As used in the present application, the expression “substantially as shown in figure . . . ” for defining the figures is intended to mean that, in view of acceptable deviations in the art, a person skilled in the art would consider that the figures are the same as the reference figures. Such deviations may be caused by known factors in the art related to instruments, operating conditions, human factors, etc. For example, a person skilled in the art would have appreciated that an endothermic start temperature and an endothermic peak temperature measured by differential scanning calorimetry (DSC) can vary significantly with experiments. In some embodiments, it is considered that two patterns are substantially identical when the change in the positions of characteristic peaks of the two patterns does not exceed ±5%, ±4%, ±3%, ±2% or ±1%. For example, it would have readily occurred to a person skilled in the art to identify whether two X-ray diffraction patterns or two DSC patterns are substantially identical. In some embodiments, it is considered that X-ray diffraction patterns are substantially identical when the change in the 2θ angle of characteristic peaks of the two X-ray diffraction patterns does not exceed ±0.3°, ±0.2° or ±0.1°.
As used in the present application, the term “approximately” should be understood to be within a range of normal tolerance in the art, for example, “approximately” can be understood to be within ±10%, ±9%, ±8%, ±7%, ±6%, ±5%, ±4%, ±3%, ±2%, ±1%, ±0.5%, ±0.1%, ±0.05% or ±0.01% of the value. Unless otherwise obvious from the context, all numeric values provided by the present application are modified with the term “approximately”.
As used in the present application, the term “pharmaceutically acceptable carrier or excipient” refers to a diluent, adjunct or vehicle that is administered with a therapeutic agent and is, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without undue toxicity, irritation, allergic response, or other problems or complications, commensurate with a reasonable benefit/risk ratio.
According to the salt of compound A, or the hydrate, solvate or crystal form of the salt thereof of the present application, the specific dosage and use method for different patients depend on many factors, including the age, weight, gender, natural health status and nutritional status of a patient, the active intensity, taking time and metabolic rate of the compound, the severity of a disorder, and the subjective judgment of a diagnosing and treating physician. A dosage of 0.01-1000 mg/kg body weight/day is preferably used here.
The structure of the crystal form of the present application can be analysed by using various analytical techniques known to a person skilled in the art, including but not limited to X-ray powder diffraction (XRD), differential scanning calorimetry (DSC) and/or thermogravimetry (TG).
Thermogravimetric analysis (TGA) is also called as thermogravimetry (TG).
The implementation process and beneficial effects of the present application are described in detail below through specific examples, which are intended to help readers better understand the essence and characteristics of the present application and are not intended to limit the scope of implementation of the present application.
The structure of the compound is determined by nuclear magnetic resonance (NMR) and/or mass spectrometry (MS).
NMR is determined with Bruker ADVANCE III 400; the solvent for determination is deuterated dimethyl sulfoxide (DMSO-d6), deuterated chloroform (CDCl3) and deuterated methanol (CD3OD); and the internal standard is tetramethylsilane (TMS).
MS is determined with Agilent 6120B (ESI).
HPLC is determined with Agilent 1260DAD high pressure liquid chromatograph (Zorba×SB-C18 100×4.6 mm).
For the column chromatography, Yantai Huanghai silica gel of 200-300 mesh silica gel is generally used as a carrier.
Instrument:
Unless otherwise specified in the examples, a solution refers to an aqueous solution.
Unless otherwise specified in the examples, a reaction is performed at room temperature,
Tetrahydrofuran (50 mL) and di-tert-butyl dicarbonate (10.6 g, 48.7 mmol) were successively added to known compound 1a (5.0 g, 32.5 mmol). After the addition, the mixture was warmed to 70° C., reacted for 16 h and concentrated under reduced pressure to remove tetrahydrofuran. The resulting mixture was slurried with petroleum ether (100 mL) for 1 h and then filtered. The filter cake was collected and dried to obtain compound 1b (6.1 g, 74%).
1H NMR (400 MHz, CD3OD) δ8.25 (d, 1H), 7.56 (d, 1H), 7.06 (d, 1H) ,1.52 (s, 9H).
LC-MS (ESI): m/z=255.1[M+H]+.
At room temperature, compound 1b (6.1 g, 24.0 mmol) was dissolved in anhydrous methanol (60 mL). Pd/C (2.1 g, with Pd content of 10% and water content of 50%) was added. Hydrogen was introduced. The mixture was warmed to 45° C. and reacted for 5 h. After filtration, the filtrate was concentrated to obtain compound 1c (4.3 g, 80%).
LC-MS (ESI): m/z=225.1[M+H]+.
Compound 1c (4.3 g, 19.2 mmol) was dissolved in methanol (50 mL). 2-bromopyridine-4-carboxaldehyde (3.6 g, 19.2 mmol) was added. The mixture was warmed to 70° C. and stirred for 15 h. The reaction solution was cooled to room temperature and concentrated under reduced pressure to remove methanol. Then dichloromethane (200 mL) and DDQ (5.3 g, 23.0 mmol) were successively added to the residue. After the addition, the mixture was stirred for 2 h at room temperature, and a saturated aqueous sodium carbonate solution (100 mL) was added. The resulting solution was stirred for 10 min and filtered. The filtrate was extracted with dichloromethane (200 mL×2). The combined organic phase was washed with water (100 mL), dried over anhydrous sodium sulphate and filtered. The filtrate was concentrated under reduced pressure, and then the residue was separated and purified by column chromatography (eluent: EA/PE=10%-50%) to obtain compound 1d (4.1 g, 54%).
LC-MS (ESI): m/z=392.1[M+H]+.
Methanol (25 mL), dichloromethane (25 mL), Pd(dppf)Cl2 (804.0 mg, 1.1 mmol) and triethylamine (4.24 g, 42.0 mmol) were successively added to compound 1d (4.1 g, 10.5 mmol). Carbon monoxide was introduced; and then the reaction solution was warmed to 120° C., stirred for 14 h, cooled to room temperature and then filtered. The filtrate was concentrated under reduced pressure, and then the residue was separated and purified by column chromatography (eluent: EA/PE=10%-50%) to obtain intermediate 1 (3.5 g, 90%).
1H NMR (400 MHz, CDCl3) δ8.95 (d, 1H), 8.89 (d, 1H), 8.26 (d, 1H), 7.86 (s, 1H), 7.54-7.47 (m, 2H) , 6.67 (s, 1H), 4.08 (s, 3H), 1.55 (s, 9H).
LC-MS (ESI): m/z=370.1[M+H]+.
At room temperature, compound 2a (580 mg, 2.0 mmol), 5-methyltetrazole (185 mg, 2.2 mmol) and triphenylphosphine (787 mg, 3.0 mmol) were dissolved in anhydrous THF (20 mL). The mixture was cooled to 0° C. under nitrogen protection, and then DEAD (520 mg, 3.0 mmol) was added dropwise. The mixture was allowed to naturally warm to room temperature, reacted overnight, concentrated under reduced pressure and subjected to column chromatography to obtain compound 2b (440 mg, 61.0%).
LC-MS (ESI): m/z=358.3[M+H]+.
Compound 2b was resolved by chiral HPLC to obtain compound 2c (tR=1.78 min, 200 mg, 45.5%).
Resolution conditions were as follows:
instrument: MG II preparative SFC (SFC-1); column type: ChiralCel OJ, 250×30 mm I.D., 5 μm; mobile phase: A: CO2, B: ethanol; gradient: B 15%; flow rate: 60 mL/min; back pressure: 100 bar; column temperature: 38° C.; column length: 220 nm; time cycle: about 5 min; sample preparation: 0.44 g of compound 2b was dissolved in a mixed solvent (4 mL) of dichloromethane and methanol; sample injection: 2 mL/injection.
At room temperature, compound 2c (200 mg, 0.55 mmol) was dissolved in dichloromethane (10 mL). Trifluoroacetic acid (2.5 mL) was added dropwise, and the mixture was stirred for another 2 h. The reaction solution was subjected to rotary evaporation to obtain a crude of intermediate 2 (300 mg), which was directly used in the next reaction without purification.
LC-MS (ESI): m/z=258.2[M+H]+.
Intermediate 1 (600.0 mg, 1.62 mmol) was dissolved in dichloromethane (5 mL), and trifluoroacetic acid (2 mL) was added. After the addition, the mixture was stirred for 2 h at room temperature, adjusted to pH=8-9 with a saturated aqueous sodium carbonate solution and extracted with dichloromethane (50 mL×2). The organic phases were combined, dried and filtered. The filtrate was concentrated to obtain compound 3a (396.0 mg, 90%).
LC-MS (ESI): m/z=270.1[M+H]+.
DMF (50 mL), (1S,2S)-2-fluorocyclopropanecarboxylic acid (425 mg, 4.1 mmol), HATU (2.1 g, 5.58 mmol) and DIEA (1.44 g, 11.16 mmol) were successively added to compound 3a (1.0 g, 3.71 mmol), and the mixture was stirred at room temperature for 5 h. The reaction was quenched by adding water, extracted 3 times with ethyl acetate and washed twice with saturated brine. The organic phase was dried and concentrated, and the residue was separated and purified by silica gel column chromatography (eluent: EA/PE=1/2) to obtain compound 3b (1.1 g, 83.4%).
LC-MS (ESI): m/z=356.3 [M+H]+.
At room temperature, compound 3b (1 g, 3.1 mmol) was dissolved in methanol (15 mL), and lithium hydroxide (700 mg) was dissolved in 20 mL of pure water. An aqueous solution of lithium hydroxide was added to the reaction solution. The mixture was stirred at 40° C. for 0.5 h and then adjusted to pH=6-7 with 2N hydrochloric acid. A large amount of solid was precipitated out, filtered by suction and washed with water (10 mL×3). The filter cake was dried at 50° C. to obtain compound 3c (1.0 g, 94.6%).
LC-MS (ESI): m/z=342.1 [M+H]+.
At room temperature, compound 3c (170 mg, 0.5 mmol) and DIPEA (130 mg, 1.0 mmol) were dissolved in DMF (5 mL), and then HATU (230 mg, 0.6 mmol) was added. The mixture was stirred for 3 min, and then intermediate 2 (300 mg, approximately 0.55 mmol) was added. The mixture was reacted for another 30 min at room temperature. 30 mL of water was added, and the reaction solution was extracted with ethyl acetate (30 mL×3). The organic phases were combined, washed with saturated sodium chloride (30 mL×1), dried over anhydrous sodium sulphate and concentrated under reduced pressure, and then the residue was subjected to column chromatography (DCM:MeOH=30:1-15:1) to obtain compound A (130 mg, 44.8%), which was an amorphous form as identified by XPRD.
LC-MS (ESI): m/z=581.3 [M+H]+.
1H NMR (400 MHz, CDCl3) δ8.74-8.71(m, 1H), 8.31(s, 1H), 8.12-8.10(m, 1H), 8.01(s, 1H), 7.90-7.87(m, 1H), 7.56-7.51(m,4H), 7.41-7.31(m, 3H), 5.55-5.52(m, 1H), 4.93-4.75(m, 2H), 3.91-3.88(m, 1H), 3.20-3.11(m, 1H), 2.91-2.80 (m, 2H), 2.56-2.50 (m, 3H), 1.94-1.84 (m, 2H), 1.61-1.23 (m, 5H).
At room temperature, 200 mg of the amorphous compound A was dissolved in 16.60 mL of isopropanol to obtain solution 1a; 150.94 mg of 1,5-naphthalene disulfonic acid was dissolved in 1.80 mL of isopropanol to obtain solution 1b; at room temperature with stirring, solution 1b was added dropwise to solution 1a to obtain solution 1c; solution 1c was continuously stirred, and a solid was precipitated immediately to obtain a suspension; the suspension was stirred overnight and then centrifuged; and the resulting solid was dried under vacuum at room temperature to obtain the hemi-1,5-naphthalene disulfonate of compound A. The compound was identified as a crystalline form by XRPD and named as crystal form I of hemi-1,5-naphthalene disulfonate of compound A.
1H NMR (400 MHz, DMSO-d6) δ10.48 (s, 1H), 8.92-8.85 (m, 1H), 8.84-8.77 (m, 1H), 8.25 (d, 1H), 8.18-8.11 (m, 2H), 7.95 (dd, 1H), 7.80 (d, 1H), 7.69-7.53 (m, 3H), 7.48-7.30 (m, 4H), 5.93 (dd, 1H), 5.07-4.82 m, 1H), 4.50 (d, 1H), 3.75-3.68 (m, 1H), 3.08 (t, 1H), 2.86 (d, 2H), 2.46 (d, 3H), 2.10-2.00 (m, 1H), 1.75-1.60 (m, 1H), 1.46-1.13 (m, 5H).
The crystal form I of hemi-1,5-naphthalene disulfonate of compound A obtained in example 1 is a crystalline form as identified by XRPD. Table 1 shows the XRPD peak list;
At room temperature, 100 mg of the amorphous compound A was ultrasonically dissolved in 2.40 mL of acetone to obtain a clear solution, i.e., solution 2a; 20.17 mg of hydrochloric acid was dissolved in 2.66 mL of acetone to obtain solution 2b; at room temperature with stirring, solution 2b was added dropwise to solution 2a to obtain solution 2c; solution 2c was continuously stirred overnight, and then a solid was precipitated to obtain a suspension; the suspension was centrifuged; and the resulting solid was dried under vacuum at room temperature to obtain the hydrochloride of compound A. The compound was identified as a crystalline form by XRPD and named as crystal form I of hydrochloride of compound A.
1H NMR (400 MHz, DMSO-d6) δ10.54 (s, 1H), 8.80 (t, 1H), 8.25 (d, 1H), 8.19-8.10 (m, 2H), 7.80 (d, 1H), 7.67-7.53 (m, 3H), 7.47-7.28 (m, 3H), 5.93 (dd, 1H), 5.10-4.85 (m, 1H), 4.50 (d, 1H), 3.72 (d, 1H), 3.08 (t, 1H), 2.89-2.79 (m, 2H), 2.45 (d, 3H), 2.07-1.93 (m, 1H), 1.75-1.60 (m, 1H), 1.44-1.10 (m, 5H).
The crystal form I of hydrochloride of compound A obtained in example 2 is a crystalline form as identified by XRPD. Table 2 shows the XRPD peak list;
At room temperature, 100 mg of the amorphous compound A was ultrasonically dissolved in 2.40 mL of acetone to obtain a clear solution, i.e., solution 3a; 40.50 mg of hydrobromic acid was dissolved in 2.33 mL of acetone to obtain solution 3b; at room temperature with stirring, solution 3b was added dropwise to solution 3a to obtain solution 3c; solution 3c was continuously stirred for 1 h, and then a solid was precipitated to obtain a suspension; the suspension was stirred and then centrifuged; and the resulting solid was dried under vacuum at room temperature to obtain the hydrobromide of compound A. The compound was identified as a crystalline form by XRPD and named as crystal form I of hydrobromide of compound A.
At room temperature, 30 mg of the amorphous compound A was ultrasonically dissolved in 0.2 mL of acetone to obtain a clear solution, i.e., solution 3a-1; 12.32 mg of hydrobromic acid was dissolved in 0.7 mL of acetone to obtain solution 3b-1; at room temperature with stirring, solution 3b-1 was added dropwise to solution 3a-1 to obtain solution 3c-1; solution 3c-1 was continuously stirred for a few minutes, and then a solid was precipitated to obtain a suspension; the suspension was stirred overnight and then centrifuged; and the resulting solid was dried under vacuum at room temperature to obtain the hydrobromide of compound A. The compound was identified as a crystalline form by XRPD and named as crystal form II of hydrobromide of compound A.
1H NMR (400 MHz, DMSO-d6) δ10.50 (s, 1H), 8.81 (t, 1H), 8.25 (d, 1H), 8.19-8.10 (m, 2H), 7.80 (d, 1H), 7.69-7.51 (m, 3H), 7.46-7.27 (m, 3H), 5.93 (dd, 1H), 5.10-4.86 (m, 1H), 4.50 (d, 1H), 3.72 (d, 1H), 3.08 (t, 1H), 2.86 (d, 2H), 2.45 (d, 3H), 2.08-1.98 (m, 1H), 1.75-1.61 (m, 1H), 1.45-1.10 (m, 5H).
The crystal form I of hydrobromide of compound A obtained in example 3 is a crystalline form as identified by XRPD. Table 3 shows the XRPD peak list;
The crystal form II of hydrobromide of compound A obtained in example 3 is a crystalline form as identified by XRPD. Table 4 shows the XRPD peak list; and
At room temperature, approximately 100 mg of the amorphous compound A was dissolved in 0.67 mL of tetrahydrofuran to obtain solution 4a; 40.27 mg of 2-naphthalenesulfonic acid was dissolved in 2.16 mL of tetrahydrofuran to obtain solution 4b; at room temperature with stirring, solution 4b was added dropwise to solution 4a to obtain solution 4c; solution 4c was continuously stirred for a few minutes, and then a solid was precipitated to obtain a suspension; the suspension was stirred overnight and then centrifuged; and the resulting solid was dried under vacuum at room temperature to obtain the 2-naphthalenesulfonate of compound A. The compound was identified as a crystalline form by XRPD and named as crystal form I of 2-naphthalenesulfonate of compound A.
At room temperature, 30 mg of the amorphous compound A was ultrasonically dissolved in 0.20 mL of 1,4-dioxane to obtain a clear solution, i.e., solution 4d; 12.0 mg of 2-naphthalenesulfonic acid was ultrasonically dissolved in 1.00 mL of 1,4-dioxane to obtain a clear solution, i.e., solution 4e; at room temperature with stirring, solution 4e was added dropwise to solution 4d to obtain solution 4f; solution 4f was continuously stirred, and then a solid was precipitated to obtain a suspension; the suspension was stirred overnight and then centrifuged; and the resulting solid was dried under vacuum at room temperature to obtain the 2-naphthalenesulfonate of compound A. The compound was identified as a crystalline form by XRPD and named as crystal form I of 2-naphthalenesulfonate of compound A.
At room temperature, 30 mg of the amorphous compound A was ultrasonically dissolved in 0.20 mL of tetrahydrofuran to obtain a clear solution, i.e., solution 4a-1; 12.10 mg of 2-naphthalenesulfonic acid was dissolved in 0.65 mL of tetrahydrofuran to obtain solution 4b-1; at room temperature with stirring, solution 4b-1 was added dropwise to solution 4a-1 to obtain solution 4c-1; solution 4c-1 was continuously stirred for 2 h, and then a solid was precipitated to obtain a suspension; the suspension was stirred overnight and then centrifuged; and the resulting solid was dried under vacuum at room temperature to obtain the 2-naphthalenesulfonate of compound A. The compound was identified as a crystalline form by XRPD and named as crystal form II of 2-naphthalenesulfonate of compound A.
1H NMR (400 MHz, DMSO-d6) δ10.48 (s, 1H), 8.84-8.77 (m, 1H), 8.24 (d, 1H), 8.19-8.10 (m, 3H), 7.96 (t, 1H), 7.93-7.84 (m, 2H), 7.80 (d, 1H), 7.72 (dd, 1H), 7.66-7.49 (m, 5H), 7.47-7.28 (m, 3H), 5.93 (d, 1H), 5.05-4.85 (m, 1H), 4.50 (d, 1H), 3.72 (d, 1H), 3.08 (t, 1H), 2.86 (d, 2H), 2.45 (d, 3H), 2.12-1.94 (m, 1H), 1.76-1.60 (m, 1H), 1.45-1.00 (m, 5H).
The crystal form I of 2-naphthalenesulfonate of compound A obtained in example 4 is a crystalline form as identified by XRPD. Table 5 shows the XRPD peak list;
The crystal form I of 2-naphthalenesulfonate of compound A obtained in example 4 is a crystalline form as identified by XRPD. Table 6 shows the XRPD peak list; and
At room temperature, 90 mg of the amorphous compound A was dissolved in 7.50 mL of isopropanol to obtain solution 5a; 30.0 mg of benzenesulfonic acid was dissolved in 0.48 mL of isopropanol to obtain solution 5b; at room temperature with stirring, solution 5b was added dropwise to solution 5a to obtain solution 5c; solution was continuously stirred for 1 h, and then a solid was precipitated to obtain a suspension; the suspension was stirred overnight and then centrifuged; and the resulting solid was dried under vacuum at room temperature to obtain the benzenesulfonate of compound A.
1H NMR (500 MHz, DMSO-d6) δ10.50 (s, 1H), 8.80 (t, 1H), 8.25 (d, 1H), 8.19-8.11 (m, 2H), 7.80 (d, 1H), 7.67-7.53 (m, 5H), 7.46-7.25 (m, 6H), 5.93 (dd, 1H), 5.02-4.74 (m, 1H), 4.49 (s, 1H), 3.71 (d, 1H), 3.07 (t, 1H), 2.84 (d, 2H), 2.46 (d, 3H), 2.09-1.98 (m, 1H), 1.76-1.61 (m, 1H), 1.45-1.11 (m, 5H).
At room temperature, 90 mg of the amorphous compound A was dissolved in 0.60 mL of acetone to obtain solution 6a; 18.0 mg of methanesulfonic acid was dissolved in 1.80 mL of acetone to obtain solution 6b; at room temperature with stirring, solution 6b was added dropwise to solution 6a to obtain solution 6c; solution 6c was continuously stirred for a few minutes, and then a solid was precipitated to obtain a suspension; the suspension was stirred overnight and then centrifuged; and the resulting solid was dried under vacuum at room temperature to obtain the methanesulfonate of compound A.
1H NMR (500 MHz, DMSO-d6) δ10.50 (s, 1H), 8.80 (t, 1H), 8.25 (d, 1H), 8.18-8.11 (m, 2H), 7.80 (d, 1H), 7.66-7.53 (m, 3H), 7.46-7.29 (m, 3H), 5.93 (dd, 1H), 5.05-4.85 (m, 1H), 4.49 (s, 1H), 3.71 (d, 1H), 3.07 (t , 1H), 2.84 (d, 2H), 2.45 (d, 3H), 2.38 (s, 3H), 2.11-2.00 (m, 1H), 1.72-1.62 (m, 1H), 1.44-1.11 (m, 5H).
At room temperature, approximately 90 mg of the amorphous compound A was dissolved in 0.60 mL of acetone to obtain solution 7a; 29.58 mg of p-toluenesulfonic acid was dissolved in 2.04 mL of acetone to obtain solution 7b; at room temperature with stirring, solution 7b was added dropwise to solution 7a to obtain solution 7c; solution 7c was continuously stirred, and then a solid was precipitated to obtain a suspension; the suspension was stirred overnight and then centrifuged; and the resulting solid was dried under vacuum at room temperature to obtain the p-toluenesulfonate of compound A.
1H NMR (500 MHz, DMSO-d6) δ10.50 (s, 1H), 8.80 (t, 1H), 8.25 (d, 1H), 8.18-8.11 (m, 2H), 7.80 (d, 1H), 7.65-7.59 (m, 2H), 7.56 (d, 1H), 7.51-7.45 (m, 2H), 7.45-7.30 (m, 3H), 7.12 (d, 2H), 5.93 (dd, 1H), 5.02-4.73 (m, 1H), 4.49 (s, 1H), 3.71 (d, 1H), 3.07 (t, 1H), 2.84 (d, 2H), 2.46 (d, 3H), 2.29 (s, 3H), 2.08-2.00 (m, 1H), 1.72-1.62 (m, 1H), 1.46-1.07 (m, 5H).
With reference to the methods in Examples 1-7, according to the feeding ratios in Table 7, the corresponding salts were prepared.
Test results: no salt form or stable salt form was prepared.
The properties of various salts of compound A are shown in Table 8.
Samples: crystal form I of hemi-1,5-naphthalene disulfonate of compound A in Example 1, crystal form I of hydrochloride of compound A in Example 2, crystal form I of hydrobromide of compound A in Example 3, and crystal form I of 2-naphthalenesulfonate of compound A in Example 4.
Experiments: the crystal form I of hemi-1,5-naphthalene disulfonate of compound A in Example 1, crystal form I of hydrochloride of compound A in Example 2, crystal form I of hydrobromide of compound A in Example 3, and crystal form I of 2-naphthalenesulfonate of compound A in Example 4 were respectively placed at an accelerated condition (open, 40° C., 75% RH (relative humidity)), a light condition (open, 25° C., total illuminance not less than 1.2×106 Lux·hr, near-ultraviolet energy not less than 200 w·hr/m2), a high humidity condition (open, drying oven) and a long-term condition (open, 25° C.-60% RH) for stability experiments. On day 0, day 5, day 10 and day 15, the samples were taken and detected for purity by HPLC (expressed as a percentage). The experimental results are shown in Table 9.
Conditions for detecting purity by HPLC: chromatographic column: ChromCore 120 C18 5 μm (4.6 mm*100); column temperature: 35° C.; detection wavelength: 220 nm; mobile phase: 10 mmol/L ammonium formate aqueous solution: acetonitrile=55:45 (v/v); flow rate: 1.0 mL/min.
Conclusion: after placed at a high temperature condition, a long-term condition or a light condition for 10 days, the compounds of Examples 1-4 have basically no change in purity and little change in single impurity content, indicating good stability; in addition, a stability test at an accelerated condition was performed on the crystal form I of 2-naphthalenesulfonate of compound A in Example 4, which showed good stability.
Test objective: by giving test compounds to SD rats via single-dose intravenous and intragastric administration and measuring the concentrations of the test compounds in plasma of rats, the pharmacokinetic characteristics and bioavailability of the test compounds in rats were evaluated.
Test compound: compound A, crystal form I of hemi-1,5-naphthalene disulfonate of compound A in Example 1 and crystal form I of 2-naphthalenesulfonate of compound A in Example 4.
Test animal: male SD rats, about 220 g, 6-8 weeks old, 6 rats/compound, purchased from Chengdu Ddossy Experimental Animals Co., Ltd.
Test method: on the day of the test, 6 SD rats were randomly grouped according to their body weight; the animals were fasted with water available for 12 to 14 h one day before the administration, and were fed 4 h after the administration; and the administration was performed according to Table 10.
Before and after the administration, 0.1 mL of blood samples were drawn from the orbits of the animals under isoflurane anaesthesia, and placed in an EDTAK2 centrifuge tube. Centrifugation was performed at 5000 rpm at 4° C. for 10 min, and plasma was collected.
Time points for sample collection in G1 and G2 groups comprise 0, 5 min, 15 min, 30 min, 1 h, 2 h, 4 h, 6 h, 8 h and 24 h.
Before analysis and detection, all samples were stored at −80° C.
30 μL of each of plasma samples, standard curve samples and quality control samples was taken, and 200 μL of acetonitrile solution containing an internal standard was added. The resulting mixture was homogeneously mixed by vortex and centrifuged at 4° C. at 12000 rpm for 10 min. 170 μL of the supernatant was taken and placed to a 96-well plate, and LC-MS/MS analysis was performed, wherein the sample size was 0.2 μL.
The main pharmacokinetic parameters were analysed by a non-compartmental model using WinNonlin 8.0 software. The test results were as shown in Table 11.
Conclusion: the salt forms of the compounds of the present application, such as crystal form I of hemi-1,5-naphthalene disulfonate of compound A in Example 1 and crystal form I of 2-naphthalenesulfonate of compound A in Example 4, have good pharmacokinetics in rats and significantly improved bioavailability compared with compound A in a free state.
Test objective: by giving test compounds to ferrets via single-dose intragastric administration, collecting plasma at different time points and measuring the concentrations of the test compounds in plasma of ferrets, the absorption of the test compounds in ferrets was evaluated in this test.
Test compound: crystal form I of hemi-1,5-naphthalene disulfonate of compound A in Example 1.
Test animal: 6 healthy adult male ferrets, about 800-1500 g, 6-10 months old, purchased from Wuxi Sangosho Biotechnology Co., Ltd.
Test method: the male ferrets selected for the test were fasted with water available for 12-14 h one day before the administration, and were fed 4 h after the administration; and the administration was performed according to Table 12.
Time points for blood collection: before the administration and 0.0833 h, 0.25 h, 0.5 h, 1 h, 2 h, 4 h, 6 h, 8 h and 24 h after the administration (10 time points in total), venous blood samples were collected into centrifuge tubes and centrifuged within 30 minutes, and the centrifuged plasma samples were stored in a refrigerator at −80° C. for PK analysis.
The test results were as shown in Table 13.
Conclusion: the salt forms of the compounds of the present application, such as crystal form I of hemi-1,5-naphthalene disulfonate of compound A in Example 1, have good pharmacokinetics in ferrets.
| Number | Date | Country | Kind |
|---|---|---|---|
| 202110157185.7 | Feb 2021 | CN | national |
| Filing Document | Filing Date | Country | Kind |
|---|---|---|---|
| PCT/CN2022/074368 | 1/27/2022 | WO |
