Salt form and crystal form of complement factor b inhibitor and preparation method and use thereof

Abstract

This invention belongs to the pharmaceutical field, specifically relating to a pharmaceutically acceptable salt form and crystal form of a compound of formula I, as well as its preparation method and application:

Description

[0001] This invention claims priority to two earlier applications: patent application No. 202210096073.X, filed with the China National Intellectual Property Administration on January 26, 2022, entitled "Salt Form, Crystal Form and Preparation Method Thereof and Application of Complement Factor B Inhibitor," and patent application No. 202211104894.X, filed with the China National Intellectual Property Administration on September 9, 2022, also entitled "Salt Form, Crystal Form and Preparation Method Thereof and Application of Complement Factor B Inhibitor." The entire contents of the earlier applications are incorporated herein by reference. Technical Field

[0002] This invention belongs to the pharmaceutical field, specifically relating to the salt form, crystal form, preparation method, and application of complement factor B inhibitors. Background Technology

[0003] Complement is a class of soluble pattern recognition molecules in the immune system that perform a variety of effector functions. Under natural conditions, complement components exist in inactive zymogen form. Various specific and non-specific immunological mechanisms break down these inactive zymogens, producing active large and small fragments. The large fragments typically remain on the surface of pathogens or cells, causing their lysis or accelerating their clearance; the small fragments leave the cell surface and mediate various inflammatory responses. Complement activation consists of two closely related processes, forming a cascade of complement activation. Currently known complement activation pathways mainly include three: the classical pathway, the lectin pathway, and the alternative pathway. Although the initiation mechanisms and activation sequences of these three complement activation pathways differ, they share a common terminal pathway. The activation of the alternative pathway is independent of the antigen-antibody complex. Typically, C3b deposited on the cell surface binds to factor B, becoming readily broken down by factor D in serum. During this process, factor B is broken down into Ba and Bb. C3b and Bb then form a complex, becoming the C3 convertase C3bBb in the alternative pathway. In this process, complement factor B plays an early and central role in the activation of the alternative pathway within the complement cascade. Here, C3b is both a product of C3 convertase breakdown and a component of the alternative pathway C3 convertase, thus forming a feedback amplification mechanism where the classical and alternative pathways influence each other. Current research has found a correlation between various diseases, including hematologic, autoimmune, inflammatory, and neurodegenerative diseases, and complement system dysfunction.

[0004] Paroxysmal nocturnal hemoglobinuria (PNH) is a chronic disease characterized by persistent hemolysis. It is a non-malignant clonal disease caused by acquired somatic PIG-A gene mutations in one or more hematopoietic stem cells and is classified as an extremely rare blood disorder (Medicine (Baltimore) 1997, 76(2):63-93). The course of the disease can manifest as varying degrees of hemolysis exacerbation (paroxysmal), chronic or recurrent acute intravascular hemolysis or subsequent venous / arterial thrombosis, ultimately leading to progressive end-organ damage and death. Most patients often present with atypical symptoms, insidious onset, prolonged course, and varying degrees of severity.

[0005] There are more than ten proteins on the surface of erythrocytes that inhibit complement pathway activation. All of these proteins are anchored to their cell membranes via glycosylphosphatidylinositol (GPI), collectively known as GPI-anchored proteins (APs). Currently, the pathogenesis of polycythemia vera (PNH) is believed to begin with hematopoietic stem cells mutating under certain conditions to produce PNH clones deficient in glycosylphosphatidylinositol (GPI). Then, due to certain factors (currently widely believed to be immune factors), hematopoietic function is impaired or fails, and the PNH clones gain proliferative advantage, exceeding that of normal clones. The presence of multiple antigens associated with GPIs also complicates the explanation of the biological behavior of PNH cells. Among the most important are CD55, the C3 convertase decay accelerator, and CD59, the membrane attack complex (MAC) inhibitor, which are closely related to the pathogenesis, clinical manifestations, diagnosis, and treatment of PNH (Frontiers in Immunology 2019, 10, 1157). CD59 can prevent C9 incorporation into the C5b-8 complex, thus preventing the formation of membrane attack units and inhibiting the terminal complement attack response. Currently, it is believed that the typical manifestations of PNH—intravascular hemolysis and thrombosis—are caused by CD59 deficiency. Patients with congenital CD59 deficiency have been reported to exhibit numerous typical symptoms of PNH, such as intravascular hemolysis, hemoglobinuria, and venous thrombosis. In PNH patients, due to GPI synthesis defects, CD59 cannot bind to the cell membrane of erythrocytes, resulting in the loss of its function in inhibiting complement pathway activation. Therefore, abnormal activation of the complement pathway occurs, attacking erythrocytes and leading to various clinical manifestations such as intravascular hemolysis, hemoglobinuria, and smooth muscle dysfunction. Currently, apart from hematopoietic stem cell transplantation to restore normal hematopoietic function, there are no other effective curative treatments for PNH. Because hematopoietic stem cell transplantation carries certain risks, and PNH is a benign clonal disease, controlling hemolytic episodes remains the main strategy for clinical treatment. Currently, only eculizumab is approved for the treatment of PNH. However, many patients still experience anemia after eculizumab treatment, and many still require continuous blood transfusions. Furthermore, eculizumab requires intravenous injection. Therefore, the development of novel inhibitors of the complement pathway for the treatment of PNH is of great significance.

[0006] IgAN is the most common primary glomerulonephritis, characterized by IgA deposition in the mesangial area as shown by immunofluorescence. The clinical manifestations are diverse, typically presenting as recurrent microscopic or gross hematuria. Current data indicate that IgAN is associated with congenital or acquired immune dysregulation. IgA1 synthesis in the mucosa increases due to irritation from viruses, bacteria, and food proteins on the respiratory or digestive tract, or IgA1-containing immune complexes deposit in the mesangial area, activating the complement bypass pathway and causing glomerular damage. Human IgA molecules are divided into two subtypes: IgA1 and IgA2. IgA1 is the predominant form circulating in the blood of healthy individuals (approximately 85%) and is also the main component deposited in the glomerular mesangial area of ​​IgAN patients. IgA molecules can exist in both monomeric and polymeric forms. The IgA1 molecule possesses a unique heavy-chain hinge region between the first and second constant regions, which can serve as a domain for O-linked glycan groups. Recent studies have found that the IgA molecules deposited in the serum and glomerular mesangial area of ​​IgAN patients are mainly glycosylated defective IgA1 (gd-IgA1). It is currently believed that the initiation step of the pathogenesis of IgAN is the abnormal increase in the production of gd-IgA1.

[0007] Over 90% of IgAN patients have complement C3 deposition in the glomerular mesangial area. 75%-100% of IgAN patients have properin and co-deposition of IgA and C3 in their renal tissue, while 30%-90% have co-deposition of complement factor H, IgA, and C3. In addition to renal tissue deposition, some studies have found that the levels of markers of the complement alternative pathway in the plasma of IgAN patients are also related to the activity of IgAN (J Nephrol 2013, 26(4):708-715). Studies have confirmed that C3a in renal tissue and urine, as well as C3a receptors in renal tissue, are significantly correlated with the activity and severity of renal damage (J clinImmunol 2014, 34(2):224-232). Other studies have confirmed that IgA can activate the complement alternative pathway under in vitro conditions. In this process, abnormalities in the IgA hinge region do not play a decisive role, but the formation of IgA polymers is the key link (Eur J Immunol 1987,17(3):321-326). Currently, complement C3 deposition in the glomerular mesangial area has become an auxiliary diagnostic marker of IgAN. A study conducted immunofluorescence detection of C3c and C3d in the renal tissue of 163 IgAN patients. The results showed that IgAN patients with higher C3c deposition intensity than C3d deposition intensity had lower glomerular filtration rate, higher incidence of glomerular capillary proliferation, and more severe hematuria, indicating that glomerular C3c deposition is related to the active pathology of IgAN (Am J Nephrol.2000,20(2):122-128). Currently, there is no specific drug treatment for IgAN in clinical practice. The main treatments are general drugs such as renin-angiotensin inhibitors (ACEIs or ARBs), glucocorticoids, and various immunosuppressants. In addition, the safety of these drugs is also a concern. For example, although glucocorticoids can reduce proteinuria, the STOP-IgAN and TESTING-I trials have clearly demonstrated the potential side effects of glucocorticoids (IgA nephropathy 2019, 95, 4, 750-756).

[0008] Arthritis is a common chronic disease, an inflammatory condition caused by inflammation, infection, degeneration, trauma, or other factors. Clinical manifestations include redness, swelling, heat, pain, impaired function, and joint deformities. It often causes severe pain, limited mobility, and body deformities, and in severe cases, can lead to disability and significantly impact quality of life. Studies have found that K / BxN mouse serum does not induce arthritis in complement factor B-deficient mice, while wild-type mice developed arthritis induced by K / BxN mouse serum (Immunity, 2002, 16, 157–168). This indicates that the complement system plays a crucial pathogenic role in the K / BxN mouse serum-induced arthritis model, and complement factor B is a potential therapeutic target for arthritis.

[0009] Other diseases associated with complement cascade include membranous nephropathy (MN), C3 glomerulonephritis (C3G), age-related macular degeneration (AMD), geographic atrophy (GA), atypical hemolytic uremic syndrome (aHUS), hemolytic uremic syndrome (HUS), hemodialysis complications, hemolytic anemia or hemodialysis, neuromyelitis (NMO), liver inflammation, inflammatory bowel disease, dermatomyositis and amyotrophic lateral sclerosis, myasthenia gravis (MG), respiratory diseases, and cardiovascular diseases.

[0010] Currently, there are no small molecule drugs that are complement factor B inhibitors for clinical treatment. Known and investigational projects include: IONIS Pharmaceuticals Inc.'s oligonucleotide drugs, developed as specific inhibitors of complement factor B (CFB) to treat, prevent, or alleviate diseases associated with dysregulation of the alternative complement pathway (WO2015038939); Novartis AG's small molecule complement factor B inhibitors for the treatment of age-related macular degeneration (AMD) and other diseases (WO2013164802, WO2013192345, WO2014143638, WO2015009616, WO2015066241), or for the treatment of diseases such as C3G and IgAN (WO2019043609A1); and Achillion Pharmaceuticals Inc.'s small molecule complement factor B inhibitors for the treatment of age-related macular degeneration (AMD) and other diseases (WO2018005552).

[0011] Inflammation and immune-related diseases are characterized by their diversity and difficulty in treatment. For PNH (progressive neonatal influenza), only eculizumab is currently available, but its high price places a heavy burden on patients. Furthermore, many patients still experience anemia after eculizumab treatment, and many still require continuous blood transfusions. In addition, eculizumab requires intravenous administration. Some diseases, such as IgAN (anti-inflammatory nephropathy), currently lack specific treatments. There are unmet clinical needs in these areas, necessitating the development of new small-molecule drugs for medical treatment.

[0012] Therefore, there is a need to develop a pharmaceutically acceptable active ingredient that is highly effective, low in toxicity, and / or long-lasting to improve the aforementioned technical problems. Summary of the Invention

[0013] To address the aforementioned technical problems, this invention provides a pharmaceutically acceptable salt of a compound of formula I:

[0014]

[0015] The pharmaceutically acceptable salt is a salt formed by a compound of formula I with an acid or base, preferably a salt formed by a compound of formula I with an acid.

[0016] According to embodiments of the present invention, the acid may be selected from inorganic or organic acids, such as hydrochloric acid, hydrofluoric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, pyrosulfuric acid, phosphoric acid, nitric acid, formic acid, acetic acid, acetoacetic acid, pyruvic acid, trifluoroacetic acid, propionic acid, butyric acid, hexanoic acid, heptanoic acid, undecanoic acid, lauric acid, benzoic acid, salicylic acid, 2-(4-hydroxybenzoyl)benzoic acid, camphoric acid, cinnamic acid, cyclopentanepropionic acid, digluconic acid, 3-hydroxy-2-naphthoic acid, nicotinic acid, pyruvic acid, pectinic acid, persulfate, 3 -Phenylacetic acid, picric acid, tervaline, 2-hydroxyethanesulfonic acid, itaconic acid, aminosulfonic acid, trifluoromethanesulfonic acid, dodecyl sulfuric acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, methanesulfonic acid, 2-naphthalenesulfonic acid, naphthalenedisulfonic acid, camphorsulfonic acid, citric acid, L-tartaric acid, stearic acid, lactic acid, oxalic acid, malonic acid, succinic acid, malic acid, adipic acid, alginic acid, maleic acid, fumaric acid, D-gluconic acid, mandelic acid, ascorbic acid, glucohepanoic acid, glycerophosphate, aspartic acid, sulfosalicylic acid, hemisulfonic acid, or thiocyanate. As an example, the acid may be selected from hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, nitric acid, methanesulfonic acid, p-toluenesulfonic acid, fumaric acid, maleic acid, citric acid, L-tartaric acid, oxalic acid, formic acid, acetic acid, trifluoroacetic acid, lauric acid, benzoic acid, and benzenesulfonic acid.

[0017] According to an embodiment of the present invention, the alkali may be selected from inorganic alkalis, such as alkali metal hydroxides or alkaline earth metal hydroxides, preferably sodium hydroxide or potassium hydroxide.

[0018] According to a preferred embodiment of the present invention, the pharmaceutically acceptable salt of the compound of Formula I is selected from one of its hydrochloride, sulfate, phosphate, methanesulfonate, p-toluenesulfonate, fumarate, maleate, citrate, L-tartrate, and oxalate.

[0019] According to a more preferred embodiment of the present invention, the pharmaceutically acceptable salt of the compound of formula I is a salt formed by the compound of formula I and hydrochloric acid, that is, the hydrochloride salt of the compound of formula I, preferably the monohydrochloride salt of the compound of formula I.

[0020] According to embodiments of the present invention, in pharmaceutically acceptable salts of the compound of formula I, the molar ratio of the compound of formula I to the acid or base can be independently selected from 1:1, 2:1, or 3:1, provided that the ions of the compound of formula I in the salt are in charge balance with the ions of the acid or base. For example, when the acid (such as hydrochloric acid, methanesulfonic acid, p-toluenesulfonic acid) has 1 ionizable hydrogen atom, the molar ratio of the compound of formula I to the acid is 1:1; when the acid (such as sulfuric acid, fumaric acid, maleic acid, citric acid, L-tartaric acid, oxalic acid) has 2 ionizable hydrogen atoms, the molar ratio of the compound of formula I to the acid can be 1:1 or 2:1; when the acid (such as phosphoric acid) has 3 ionizable hydrogen atoms, the molar ratio of the compound of formula I to the acid is 1:1, 2:1, or 3:1.

[0021] The present invention also provides a method for preparing a pharmaceutically acceptable salt of a compound of formula I, the method comprising reacting a compound of formula I with an acid or base to prepare a pharmaceutically acceptable salt of the compound of formula I.

[0022] According to an embodiment of the present invention, the preparation method comprises reacting a compound of formula I with the acid or base in a solvent to prepare a pharmaceutically acceptable salt of the compound of formula I.

[0023] According to embodiments of the present invention, the acid or base has the definition described above independently of each other.

[0024] According to an embodiment of the present invention, the solvent may be selected from alcohols, ketones, esters, ethers, combinations of two or more of the solvents, or mixtures of the above solvents or combinations with water.

[0025] According to embodiments of the present invention, the alcohols may be selected from alcohols having 1-8 carbon atoms, such as methanol, ethanol, n-propanol, isopropanol, n-butanol, neopentyl alcohol, or combinations thereof; the ketones may be selected from ketones having 3-10 carbon atoms, such as acetone, butanone, pentanone, methyl ethyl ketone, 4-methyl-2-pentanone, or combinations thereof; the esters may be selected from organic carboxylic acid esters, such as methyl formate, ethyl acetate, isobutyl formate, ethyl propyl acetate, or combinations thereof; and the ethers may be straight-chain or branched alkyl ethers or cyclic ether compounds, such as methyl tert-butyl ether, tetrahydrofuran, 2-methyl-tetrahydrofuran, or combinations thereof.

[0026] According to an embodiment of the present invention, the molar ratio of the compound of formula I to the acid or base can be 1:0.8 to 1:1.5, preferably 1:0.9 to 1:1.3, and more preferably 1:1.0 to 1:1.1.

[0027] According to an embodiment of the present invention, in the preparation method, the reaction temperature can be selected within a wide range, for example, 20℃ to 80℃, preferably 30℃ to 60℃.

[0028] According to an embodiment of the present invention, the preparation method further includes a step of filtration and / or drying after the reaction is completed, to prepare a pharmaceutically acceptable salt of the compound of formula I.

[0029] According to an embodiment of the present invention, in the preparation method, the drying temperature can be selected within a wide range, for example, 20℃ to 80℃, preferably 30℃ to 60℃.

[0030] According to an embodiment of the present invention, in the step preparation method, the drying pressure can be 0-20 kPa, preferably 0-10 kPa, and more preferably 5-10 kPa.

[0031] The present invention also provides crystals of a single hydrochloride salt of compound I, preferably single crystals. The unit cell parameters of the single crystal are as follows:

[0032] Orthorhombic crystal system, space group P212121;

[0033]

[0034]

[0035]

[0036]

[0037] Z = 4.

[0038] The present invention also provides crystals of the monohydrochloride salt of the compound of formula I, and in particular a method for preparing a single crystal thereof, comprising dissolving the monohydrochloride salt of the compound of formula I in solvent A and then placing it in an atmosphere of solvent B for diffusion.

[0039] Solvent A can be an alcohol solvent, such as a combination of two or more of methanol and ethanol.

[0040] Solvent B can be an ester solvent, an ether solvent, or a combination of two or more thereof. The ester solvent can be selected from organic carboxylic acid esters, such as methyl formate, ethyl acetate, isobutyl formate, ethyl propyl acetate, or a combination of two or more thereof; the ether solvent can be a straight-chain or branched alkyl ether, a cyclic ether compound, or a combination of two or more thereof, such as methyl tert-butyl ether, tetrahydrofuran, 2-methyl-tetrahydrofuran, or a combination of two or more thereof.

[0041] The present invention also provides crystalline forms of monohydrochloride salts of Formula I compounds, selected from crystal forms A, B, C, D or E as described below.

[0042] A monohydrochloride salt of a compound of formula I, wherein crystal form A exhibits characteristic peaks at 9.66±0.20°, 16.08±0.20°, and 23.46±0.20° in X-ray powder diffraction using Cu-Kα radiation and expressed in 2θ angles.

[0043] Preferably, the crystal form A is subjected to Cu-Kα radiation, and the X-ray powder diffraction, expressed in 2θ angles, has characteristic peaks at 9.66±0.20°, 16.08±0.20°, 18.10±0.20°, 21.30±0.20°, and 21.68±0.20°.

[0044] Preferably, the crystal form A, when subjected to Cu-Kα radiation, exhibits characteristic peaks in X-ray powder diffraction at angles of 2θ at 9.66±0.20°, 11.62±0.20°, 16.08±0.20°, 18.10±0.20°, 21.30±0.20°, 21.68±0.20°, 23.40±0.20°, and 25.42±0.20°.

[0045] Preferably, the crystal form A, when subjected to Cu-Kα radiation, exhibits characteristic peaks in X-ray powder diffraction at angles of 2θ at 9.66±0.20°, 11.62±0.20°, 16.08±0.20°, 16.84±0.20°, 18.10±0.20°, 19.64±0.20°, 21.30±0.20°, 21.68±0.20°, 23.40±0.20°, 24.96±0.20°, and 25.42±0.20°.

[0046] Preferably, the crystal form A, when subjected to Cu-Kα radiation, exhibits characteristic peaks in X-ray powder diffraction at a 2θ angle, as shown in Table 1, wherein the error range of the 2θ angle is ±0.20°.

[0047] Table 1. XRPD analysis data for crystal form A

[0048] Peak number 2θ[°] Relative strength % Peak number 2θ[°] Relative strength % 1 8.02 19.6 16 26.78 11.9 2 9.66 100 17 27.66 25.6 3 11.62 38.2 18 28.44 7.2 4 12.78 13.9 19 28.70 7.7 5 14.32 13.1 20 29.04 9.5 6 16.08 84.5 21 29.70 13.9 7 16.84 24 22 30.14 14.6 8 18.10 64.7 23 32.38 10.9 9 19.64 32 24 33.82 6.4 10 21.30 53.5 25 35.41 7.3 11 21.68 42.8 26 36.89 5.4 12 23.46 93.7 27 37.92 9.3 13 24.96 19 28 38.14 8.7 14 25.42 37.6 29 39.58 5 15 26.10 11.7

[0049] Preferably, the crystal form A has essentially the following characteristics: Figure 1 The powder X-ray diffraction pattern shown.

[0050] According to an embodiment of the present invention, the crystal form A is an anhydrous form of the monohydrochloride salt of compound I.

[0051] According to an embodiment of the present invention, differential scanning calorimetry (DSC) analysis of crystal form A shows that the first endothermic peak appears near the peak temperature of 192.73°C and the first exothermic peak appears near the peak temperature of 201.78°C.

[0052] Preferably, the crystal form A has essentially the following characteristics: Figure 2 The DSC diagram shown.

[0053] According to an embodiment of the present invention, thermogravimetric analysis (TGA) of crystal form A shows a weight loss of approximately 1.41% in the range of 90°C to 180°C.

[0054] Preferably, the crystal form A has essentially the following characteristics: Figure 3 The TGA diagram shown.

[0055] According to an embodiment of the present invention, crystal form A is an irregularly shaped crystal. Preferably, the grain size of crystal form A does not exceed 20 μm.

[0056] Preferably, the crystal form A has essentially the following characteristics: Figure 4 The PLM diagram shown.

[0057] A monohydrochloride salt of Formula I, crystal form B, wherein the crystal form B exhibits characteristic peaks at 18.10±0.20°, 19.80±0.20°, and 22.10±0.20° in X-ray powder diffraction using Cu-Kα radiation and expressed in 2θ angles.

[0058] Preferably, the crystal form B, when subjected to Cu-Kα radiation, exhibits characteristic peaks in X-ray powder diffraction at 9.48±0.20°, 15.44±0.20°, 18.10±0.20°, 19.80±0.20°, 22.10±0.20°, and 30.92±0.20° when expressed in 2θ angles.

[0059] Preferably, the crystal form B, when subjected to Cu-Kα radiation, exhibits characteristic peaks in X-ray powder diffraction at angles of 2θ at 9.48±0.20°, 10.78±0.20°, 15.44±0.20°, 18.10±0.20°, 19.18±0.20°, 19.80±0.20°, 22.10±0.20°, and 30.92±0.20°.

[0060] Preferably, the crystal form B, when subjected to Cu-Kα radiation, exhibits characteristic peaks in X-ray powder diffraction at a 2θ angle, as shown in Table 2, wherein the error range of the 2θ angle is ±0.20°.

[0061] Table 2 XRPD analysis data for crystal form B

[0062] Peak number 2θ[°] Relative strength % Peak number 2θ[°] Relative strength % 1 9.48 32.4 13 23.40 8.2 2 10.78 22.6 14 24.10 13.3 3 12.08 7.1 15 25.41 7.8 4 14.68 14.5 16 26.22 9 5 15.44 27.8 17 28.16 14.4 6 18.10 51.2 18 29.18 8 7 19.18 20.7 19 29.58 5.5 8 19.80 49.9 20 30.13 5.2 9 20.60 17.9 21 30.92 24.4 10 21.34 12.4 22 33.02 4.6 11 22.10 100 23 35.50 8.7 12 22.94 9.3 24 41.28 4.8

[0063] Preferably, the crystal form B has essentially the following characteristics: Figure 5 The powder X-ray diffraction pattern shown.

[0064] According to an embodiment of the present invention, the crystal form B is a hydrate of a monohydrochloride salt of compound I, such as a monohydrate of a monohydrochloride salt of compound I.

[0065] According to an embodiment of the present invention, differential scanning calorimetry (DSC) analysis of crystal form B shows that the first endothermic peak appears near the peak temperature of 85.87°C, the second endothermic peak appears near the peak temperature of 197.54°C, and the first exothermic peak appears near the peak temperature of 205.68°C.

[0066] Preferably, the crystal form B has essentially the following characteristics: Figure 6 The DSC diagram shown.

[0067] According to an embodiment of the present invention, thermogravimetric analysis (TGA) of the crystal form B shows a weight loss of about 3.42% from 21.49°C to 120°C and a weight loss of about 0.49% from 179.88°C to 207.94°C.

[0068] Preferably, the crystal form B has essentially the following characteristics: Figure 7 The TGA diagram shown.

[0069] According to an embodiment of the present invention, crystal form B is an irregularly shaped crystal. Preferably, the grain size of crystal form B does not exceed 20 μm.

[0070] Preferably, the crystal form B has essentially the following characteristics: Figure 8 The PLM diagram shown.

[0071] According to an embodiment of the present invention, crystal form A is subjected to high humidity conditions to obtain crystal form B. The high humidity conditions are preferably 40°C and 75%–95% relative humidity.

[0072] According to an embodiment of the present invention, crystal form B is dried to obtain crystal form A. The drying conditions are preferably vacuum drying at 40°C.

[0073] A crystal form C of a monohydrochloride salt of Formula I, wherein the crystal form C exhibits characteristic peaks at 14.74±0.20°, 17.80±0.20°, 20.08±0.20°, and 21.98±0.20° when X-ray powder diffraction is performed using Cu-Kα radiation and expressed in 2θ angles.

[0074] Preferably, the crystal form C, when irradiated with Cu-Kα, exhibits characteristic peaks in X-ray powder diffraction at 14.74±0.20°, 17.80±0.20°, 19.58±0.20°, 20.08±0.20°, 21.98±0.20°, 22.94±0.20°, and 25.92±0.20° in 2θ angles.

[0075] Preferably, the crystal form C, when subjected to Cu-Kα radiation, exhibits characteristic peaks in X-ray powder diffraction at angles of 2θ at 14.74±0.20°, 17.80±0.20°, 19.58±0.20°, 20.08±0.20°, 21.98±0.20°, 22.94±0.20°, 25.92±0.20°, and 33.48±0.20°.

[0076] Preferably, the crystal form C, when subjected to Cu-Kα radiation, exhibits characteristic peaks in X-ray powder diffraction at a 2θ angle, as shown in Table 3, wherein the error range of the 2θ angle is ±0.20°.

[0077] Table 3 XRPD analysis data for crystal form C

[0078]

[0079]

[0080] Preferably, the crystal form C has essentially the following characteristics: Figure 9 The powder X-ray diffraction pattern shown.

[0081] According to an embodiment of the present invention, the crystal form C is an anhydrous form of the monohydrochloride salt of compound I.

[0082] According to an embodiment of the present invention, differential scanning calorimetry (DSC) analysis of the crystal form C shows that the first endothermic peak appears near the peak temperature of 209.93°C and the first exothermic peak appears near the peak temperature of 215.80°C.

[0083] Preferably, the crystal form C has essentially the following characteristics: Figure 10 The DSC diagram shown.

[0084] According to an embodiment of the present invention, thermogravimetric analysis (TGA) of the crystal form C shows a weight loss of about 0.29% in the range of 21.62°C to 120°C and a weight loss of about 0.52% in the range of 173.94°C to 216.60°C.

[0085] Preferably, the crystal form C has essentially the following characteristics: Figure 11 The TGA diagram shown.

[0086] According to an embodiment of the present invention, crystal form C is an irregularly shaped crystal. Preferably, the grain size of crystal form C does not exceed 20 μm.

[0087] Preferably, the crystal form C has essentially the following characteristics: Figure 12 The PLM diagram shown.

[0088] A monohydrochloride salt of Formula I has a crystal form D, wherein the crystal form D exhibits characteristic peaks at 15.74±0.20°, 16.58±0.20°, 21.98±0.20°, and 23.82±0.20° when X-ray powder diffraction is performed using Cu-Kα radiation and expressed in 2θ angles.

[0089] Preferably, the crystal form D is subjected to Cu-Kα radiation, and the X-ray powder diffraction, expressed in 2θ angles, has characteristic peaks at 10.16±0.20°, 11.90±0.20°, 15.74±0.20°, 16.58±0.20°, 19.22±0.20°, 20.24±0.20°, 21.98±0.20°, and 23.82±0.20°.

[0090] Preferably, the crystal form D, when subjected to Cu-Kα radiation, exhibits characteristic peaks in X-ray powder diffraction at angles of 2θ at 10.16±0.20°, 11.90±0.20°, 12.60±0.20°, 15.74±0.20°, 16.58±0.20°, 19.22±0.20°, 19.80±0.20°, 21.98±0.20°, 22.66±0.20°, 23.18±0.20°, 23.82±0.20°, 24.94±0.20°, 26.24±0.20°, 26.80±0.20°, and 27.50±0.20°.

[0091] Preferably, the crystal form D is subjected to Cu-Kα radiation, and the X-ray powder diffraction, expressed in 2θ angles, exhibits characteristic peaks as shown in Table 4, wherein the error range of the 2θ angles is ±0.20°.

[0092] Table 4. XRPD analysis data for crystal form D

[0093] Peak number 2θ[°] Relative strength % Peak number 2θ[°] Relative strength % 1 10.16 37.6 13 23.82 43.9 2 11.90 25.6 14 24.94 11.8 3 12.60 14.6 15 25.60 9.8 4 15.74 100 16 26.24 23.3 5 16.58 43.5 17 26.80 11.7 6 19.22 40.1 18 27.50 12.9 7 19.80 13.9 19 29.36 7 8 20.24 26.4 20 29.88 13.6 9 21.12 9.8 21 31.00 6.6 10 21.98 41.1 22 32.48 4.8 11 22.66 10.4 23 36.20 6.3 12 23.18 14.4 24 36.76 4.4

[0094] Preferably, the crystal form D has essentially the following characteristics: Figure 13 The powder X-ray diffraction pattern shown.

[0095] According to an embodiment of the present invention, the crystal form D is a solvate of a monohydrochloride salt of Formula I, such as a dichloromethane solvate of a monohydrochloride salt of Formula I, or a monodichloromethane solvate of a monohydrochloride salt of Formula I (or simply a monodichloromethane solvate).

[0096] According to an embodiment of the present invention, differential scanning calorimetry (DSC) analysis of the crystal form D shows that the first exothermic peak appears near the peak temperature of 196.53°C when heated.

[0097] Preferably, the crystal form D has essentially the following characteristics: Figure 14 The DSC diagram shown.

[0098] According to an embodiment of the present invention, thermogravimetric analysis (TGA) of the crystal form D shows a weight loss of approximately 6.31% in the range of 22.07°C to 120°C.

[0099] Preferably, the crystal form D has essentially the following characteristics: Figure 15 The TGA diagram shown.

[0100] According to an embodiment of the present invention, the crystal form D is an irregularly shaped crystal. Preferably, the grain size of the crystal form C does not exceed 10 μm.

[0101] Preferably, the crystal form D has essentially the following characteristics: Figure 16 The PLM diagram shown.

[0102] A monohydrochloride salt of Formula I has a crystal form E, wherein the crystal form E exhibits characteristic peaks at 9.36±0.20°, 15.22±0.20°, 16.88±0.20°, and 22.10±0.20° when X-ray powder diffraction is performed using Cu-Kα radiation and expressed in 2θ angles.

[0103] Preferably, the crystal form E is irradiated using Cu-Kα radiation, and the X-ray powder diffraction, expressed in 2θ angles, has characteristic peaks at 7.20±0.20°, 9.36±0.20°, 15.22±0.20°, 16.88±0.20°, 21.10±0.20°, 22.10±0.20°, and 23.68±0.20°.

[0104] Preferably, the crystal form E is subjected to Cu-Kα radiation, and the X-ray powder diffraction, expressed in 2θ angles, has characteristic peaks at 7.20±0.20°, 9.36±0.20°, 15.22±0.20°, 16.88±0.20°, 18.78±0.20°, 21.10±0.20°, 22.10±0.20°, 23.68±0.20°, 26.04±0.20°, and 27.86±0.20°.

[0105] Preferably, the crystal form E is subjected to Cu-Kα radiation, and the X-ray powder diffraction, expressed in 2θ angles, exhibits characteristic peaks as shown in Table 5, wherein the error range of the 2θ angles is ±0.20°.

[0106] Table 5. XRPD analysis data for crystal form E

[0107] Peak number 2θ[°] Relative strength % Peak number 2θ[°] Relative strength % 1 7.20 35.5 12 22.10 64.1 2 9.36 100 13 22.70 13.1 3 11.32 14.4 14 23.68 34.4 4 12.98 8 15 26.04 24.2 5 15.22 58.7 16 26.90 9.7 6 16.88 59 17 27.86 27.3 7 17.90 9.5 18 29.38 11.5 8 18.78 27.7 19 30.56 10 9 19.18 23.1 20 33.28 14.8 10 19.92 20 21 37.54 8.4 11 21.10 40.3

[0108] Preferably, the crystal form E has essentially the following characteristics: Figure 17 The powder X-ray diffraction pattern shown.

[0109] According to an embodiment of the present invention, the crystal form E is a solvate of a monohydrochloride salt of Formula I, such as an isopropanol solvate of a monohydrochloride salt of Formula I.

[0110] The present invention also provides a method for preparing the crystal form of the monohydrochloride salt of compound I.

[0111] One method for preparing crystal form A includes: dissolving a compound of formula I in an alcohol solvent, adding a solution of HCl in the alcohol solvent to form a salt, and then adding n-alkane to crystallize, thereby obtaining crystal form A.

[0112] The alcohol solvent is selected from ethanol and / or isopropanol, preferably isopropanol.

[0113] The n-alkane is selected from n-hexane and / or n-heptane, preferably n-heptane.

[0114] The mass-to-volume ratio of the compound of formula I, the alcohol solvent, and the n-alkane is 1 g:(10-30) mL:(10-30) mL, preferably 1 g:(15-25) mL:(15-25) mL.

[0115] The concentration of the HCl solution in the alcohol solvent is 1-3 mol / L, for example 2 mol / L.

[0116] The heating temperature is 45-75℃, preferably 48-60℃.

[0117] Method 2 for preparing crystal form A includes: heating and stirring the monohydrochloride salt of compound I in an alcohol solvent and n-alkane until it is dissolved and then crystallizing to obtain crystal form A.

[0118] The alcohol solvent is selected from ethanol and / or isopropanol, preferably isopropanol.

[0119] The n-alkane is selected from n-hexane and / or n-heptane, preferably n-heptane.

[0120] The mass-volume ratio of the compound of formula I monohydrochloride, alcohol solvent and n-alkane is 1g:(10-30)mL:(10-30)mL, preferably 1g:(15-25)mL:(15-25)mL, for example 1g:20mL:20mL.

[0121] The heating temperature is 45-75℃, preferably 48-60℃.

[0122] According to an embodiment of the present invention, the preparation method of crystal form A in method one or two further includes the steps of cooling, filtering, and drying.

[0123] According to a preferred embodiment of the present invention, the preparation method of crystal form A includes: dissolving the compound of formula I in isopropanol, adding an isopropanol solution of HCl, mixing, adding n-heptane and stirring, filtering, and drying to obtain crystal form A.

[0124] According to a preferred embodiment of the present invention, the preparation method of crystal form A includes: adding the monohydrochloride salt of compound I to a mixed solvent of isopropanol and n-heptane, heating and stirring, cooling to room temperature, filtering, and vacuum drying to obtain crystal form A.

[0125] The mass-to-volume ratio of the compound of formula I, monohydrochloride, isopropanol, and n-heptane is 1g:(10-30)mL:(10-30)mL, for example, 1g:20mL:20mL.

[0126] A method for preparing crystal form B includes: placing crystal form A under high humidity conditions to obtain crystal form B.

[0127] According to an embodiment of the present invention, the temperature of the high humidity condition is 30-50°C, and the humidity is 60%-98%.

[0128] The preferred high humidity conditions are 40°C and 75%–95% humidity.

[0129] The preparation method of crystal form C includes: dissolving the compound of formula I in an alcohol solvent, then adding a solution of HCl in the alcohol solvent to form a salt, and then adding an ether solvent or an ester solvent to crystallize, thereby obtaining crystal form C.

[0130] According to an embodiment of the present invention, the preparation method of crystal form C includes: dissolving the compound of formula I in an alcohol solvent, adding a solution of HCl in the alcohol solvent, stirring, filtering, adding an ether solvent or an ester solvent dropwise to the filtrate, stirring, filtering, and drying to obtain crystal form C.

[0131] The alcohol solvent is selected from methanol, ethanol or isopropanol, preferably methanol.

[0132] The ether solvent is selected from dimethyl ether, diethyl ether, propyl ether, or methyl tert-butyl ether, preferably methyl tert-butyl ether.

[0133] The ester solvent is selected from ethyl acetate or isopropyl acetate.

[0134] The mass-volume ratio of the compound of Formula I, the alcohol solvent, and the ether solvent is 1g:(2-8)mL:(20-40)mL, preferably 1g:(3-6)mL:(20-30)mL, for example 1g:4mL:25mL.

[0135] The concentration of the HCl solution in the alcohol solvent is 1-3 mol / L, for example 1.5-2.5 mol / L, and exemplarily 1.8 mol / L; the mass ratio of the compound of Formula I and the HCl solution in the alcohol solvent is 1 g : (0.5-1.5) g, for example 1 g : (0.8-1.2) g.

[0136] According to a preferred embodiment of the present invention, the preparation method of crystal form C includes: dissolving the compound of formula I in methanol, adding a methanol solution of HCl, stirring, filtering, adding methyl tert-butyl ether to the filtrate, filtering, and drying to obtain crystal form C.

[0137] The preparation method of crystal form D includes: suspending and stirring the monohydrochloride salt of compound I in a haloalkane at room temperature to crystallize, thereby obtaining crystal form D.

[0138] According to an embodiment of the present invention, the preparation method of crystal form D includes: adding the monohydrochloride salt of compound I to a haloalkane, stirring, separating the resulting suspension, drying the separated solid, and obtaining the solid as crystal form D.

[0139] The haloalkane is selected from dichloromethane, trichloromethane, or carbon tetrachloride, preferably dichloromethane.

[0140] The mass-to-volume ratio of the monohydrochloride salt of the compound of formula I to the haloalkane is 1 g:(15-35) mL, preferably 1 g:(18-25) mL, for example 1 g:20 mL.

[0141] The separation is performed using known separation methods, preferably centrifugation.

[0142] The drying is performed under reduced pressure and vacuum drying under heating conditions, preferably under reduced pressure and vacuum drying at 40°C.

[0143] According to a preferred embodiment of the present invention, the method for preparing crystal form D includes: adding the monohydrochloride salt of compound I to dichloromethane, stirring at room temperature, and then separating the solid to obtain crystal form D.

[0144] The method for preparing crystal form E includes: suspending and stirring the monohydrochloride salt of compound I in an alcohol solvent at room temperature to crystallize it, thereby obtaining crystal form E.

[0145] According to an embodiment of the present invention, the monohydrochloride salt of compound I is added to an alcohol solvent, stirred, and the resulting suspension is separated. The separated solid is dried, and the resulting solid is crystal form E.

[0146] The alcohol solvent is selected from methanol, ethanol or isopropanol, preferably isopropanol.

[0147] The mass-to-volume ratio of the monohydrochloride salt of the compound of formula I to the alcohol solvent is 1g:(15-35)mL, preferably 1g:(18-25)mL, for example 1g:20mL.

[0148] The separation is performed using known separation methods, preferably centrifugation.

[0149] The drying is performed under reduced pressure and vacuum drying under heating conditions, preferably under reduced pressure and vacuum drying at 40°C.

[0150] According to a preferred embodiment of the present invention, the method for preparing crystal form E includes: adding the monohydrochloride salt of compound I to isopropanol, stirring at room temperature, and then separating the solid to obtain crystal form E.

[0151] The present invention also provides a pharmaceutical composition comprising at least one pharmaceutically acceptable salt of the compound of Formula I (e.g., the hydrochloride salt, such as crystal form A, crystal form B, crystal form C, crystal form D, and crystal form E of the hydrochloride salt), and optionally a pharmaceutically acceptable excipient. Preferably, the pharmaceutical composition is in the form of a formulation.

[0152] The present invention also provides a formulation comprising at least one of a pharmaceutically acceptable salt of the compound of Formula I, crystal form A, crystal form B, crystal form C, crystal form D, and crystal form E, and optionally a pharmaceutically acceptable excipient.

[0153] The present invention also provides the use of at least one of the pharmaceutically acceptable salts of the compound of Formula I as described above (e.g., the hydrochloride salts, such as crystal forms A, B, C, D, and E of the hydrochloride salts) or the pharmaceutical compositions thereof in the preparation of a medicament for the prevention and / or treatment of complement factor B-mediated diseases or conditions.

[0154] According to embodiments of the present invention, the complement factor B-mediated diseases or conditions are selected from at least one of the following: paroxysmal nocturnal hemoglobinuria (PNH), primary glomerulonephritis (IgAN), membranous nephropathy (MN), C3 glomerulonephritis (C3G), age-related macular degeneration (AMD), geographic atrophy (GA), atypical hemolytic uremic syndrome (aHUS), hemolytic uremic syndrome (HUS), diabetic retinopathy (DR), hemodialysis complications, hemolytic anemia or hemodialysis, neuromyelitis (NMO), arthritis, rheumatoid arthritis, liver inflammation, dermatomyositis and amyotrophic lateral sclerosis, myasthenia gravis (MG), respiratory diseases, and cardiovascular diseases.

[0155] The present invention also provides a method for the prevention and / or treatment of diseases associated with complement factor B inhibitors, comprising administering to an individual in need a therapeutically effective amount of a pharmaceutically acceptable salt of the compound I described above (e.g., the hydrochloride salt, such as crystal form A, crystal form B, crystal form C, crystal form D, crystal form E of the hydrochloride salt) or at least one of the pharmaceutical compositions.

[0156] According to embodiments of the present invention, the disease or condition associated with complement factor B inhibitor is selected from at least one of the following: paroxysmal nocturnal hemoglobinuria (PNH), primary glomerulonephritis (IgAN), membranous nephropathy (MN), C3 glomerulonephritis (C3G), age-related macular degeneration (AMD), geographic atrophy (GA), atypical hemolytic uremic syndrome (aHUS), hemolytic uremic syndrome (HUS), diabetic retinopathy (DR), hemodialysis complications, hemolytic anemia or hemodialysis, neuromyelitis (NMO), arthritis, rheumatoid arthritis, liver inflammation, dermatomyositis and amyotrophic lateral sclerosis, myasthenia gravis (MG), respiratory diseases, and cardiovascular diseases.

[0157] The treatment methods of the present invention may include administering alone a pharmaceutically acceptable salt of a compound of formula I of the present invention (e.g., the hydrochloride salt, such as crystal form A, crystal form B, crystal form C, crystal form D, crystal form E of the hydrochloride salt) or the pharmaceutical composition thereof, or in combination with one, two or more other chemotherapeutic agents. Administration of multiple drugs may be performed simultaneously or sequentially.

[0158] In the context of this specification, "above," "below," and "within" should be understood to include the stated number. As an example, "at least one" should be understood as "one, two, or more kinds." Similarly, "two or more kinds" should be understood as "two or more kinds," such as "two, three, four, or more kinds."

[0159] Beneficial effects

[0160] The salts of the compounds of formula I of this invention (especially hydrochloride, phosphate, and maleate) exhibit high stability and high water solubility, significantly enhancing absorption and bioavailability during oral administration. Furthermore, the hydrochloride crystal form of the compounds of formula I of this invention demonstrates high stability, good solubility, and low hygroscopicity, showing promising potential for drug development. In addition, the preparation methods for the salts and crystal forms of the compounds of formula I of this invention are convenient, easy to control, reproducible, and operate under mild reaction conditions, resulting in high product yields, which is beneficial for industrial production. Attached Figure Description

[0161] Figure 1 The image shows the XRPD pattern of the monohydrochloride crystal form A of compound I.

[0162] Figure 2 The image shows the DSC spectrum of the monohydrochloride crystal form A of compound I.

[0163] Figure 3 The TGA spectrum is shown for the monohydrochloride crystal form A of compound I.

[0164] Figure 4 The PLM spectrum is shown for the monohydrochloride crystal form A of compound I.

[0165] Figure 5 The image shows the XRPD pattern of the monohydrochloride crystal form B of compound I.

[0166] Figure 6 The image shows the DSC spectrum of the monohydrochloride crystal form B of compound I.

[0167] Figure 7 The TGA spectrum is for monohydrochloride crystal form B of compound I.

[0168] Figure 8 The PLM spectrum is for monohydrochloride crystal form B of compound I.

[0169] Figure 9 The image shows the XRPD pattern of the monohydrochloride crystal form C of compound I.

[0170] Figure 10 The image shows the DSC spectrum of the monohydrochloride crystal form C of compound I.

[0171] Figure 11 The TGA spectrum is shown for the monohydrochloride crystal form C of compound I.

[0172] Figure 12 The PLM spectrum is shown for the monohydrochloride crystal form C of compound I.

[0173] Figure 13 The image shows the XRPD pattern of the monohydrochloride crystal form D of compound I.

[0174] Figure 14 The image shows the DSC spectrum of the monohydrochloride crystal form D of compound I.

[0175] Figure 15 The TGA spectrum is shown for the monohydrochloride crystal form D of compound I.

[0176] Figure 16 The PLM spectrum is shown for the monohydrochloride crystal form D of compound I.

[0177] Figure 17 The image shows the XRPD pattern of the monohydrochloride crystal form E of compound I.

[0178] Figure 18 The image shows the XRPD results of compound I, monohydrochloride crystal form C, after being placed under accelerated, high-temperature conditions for one month.

[0179] Figure 19 The image shows the XRPD results of compound I, monohydrochloride crystal form C, after being exposed to high temperature and high humidity for 1 day.

[0180] Figure 20 The image shows the XRPD results of compound I, monohydrochloride crystal form C, after being exposed to high temperature and high humidity for 3 days.

[0181] Figure 21 The data (ng / mL) are experimental data of blood drug concentration curves in cynomolgus monkeys used in biological examples.

[0182] Figure 22 The data are experimental data (% relative to 0h) of serum AP activity curve of cynomolgus monkeys in the biological examples.

[0183] Figure 23 Experimental data on streptococcal-induced rheumatoid arthritis in rats, as presented in the biological examples.

[0184] Figure 24 This is a single crystal diagram of the monohydrochloride salt of compound I. Detailed Implementation

[0185] The technical solution of the present invention will be further described in detail below with reference to specific embodiments. It should be understood that the following embodiments are merely illustrative and explanatory of the present invention, and should not be construed as limiting the scope of protection of the present invention. All technologies implemented based on the above content of the present invention are covered within the scope of protection intended by the present invention.

[0186] Unless otherwise stated, the raw materials and reagents used in the following examples are commercially available products or can be prepared by known methods.

[0187] Experimental instrument parameters

[0188] X-ray powder diffraction (XRPD)

[0189] The equipment used was a Shimadzu XRD-6000, and the sample was scanned using the following parameters:

[0190] The radiation source is a Cu-Kα target.

[0191] The minimum operating voltage and current of the fluorescent tube are 40kV and 30mA, respectively.

[0192] The 2-Theta value of the sample scanning range is from 2° to 50°. The scanning speed is 5 deg / min.

[0193] Thermogravimetric analysis (TGA)

[0194] Weigh approximately 5 mg of sample into a crucible, protect it with nitrogen, and heat it from 30 °C to 300 °C at a rate of 20 °C / min. Hold the temperature at 300 °C for 1 min.

[0195] Differential Scanning Calorimeter (DSC)

[0196] Weigh approximately 1–5 mg of powder sample and place it in a sealed aluminum crucible. Make a pinhole in the crucible lid. Under nitrogen protection, perform a differential thermal scan by heating from 30°C to 300°C and holding at 300°C for 1 minute. The heating rate is 20°C / min.

[0197] Polarizing microscope (PLM)

[0198] The sample was dispersed in a medium (silicone oil), and the sample was observed using a 10X eyepiece and a 10X objective lens. Images were recorded using a camera and computer system.

[0199] Dynamic moisture adsorption (DVS)

[0200] Under a relative humidity (RH) cycle of 0%–95%–0%, approximately 10 mg of sample was weighed and subjected to a moisture absorption / desorption characteristic test at 25°C. The parameters are as follows:

[0201]

[0202] Hygroscopicity classification:

[0203]

[0204] *: Under conditions of 25±1℃ and 80±2%RH (European Pharmacopoeia 10.0)

[0205] "W": Moisture gain at 80% RH.

[0206] Single crystal testing instruments and conditions

[0207]

[0208] Explanation of Abbreviations

[0209] 40℃ / 75%RH refers to the conditions of 40℃ and 75% humidity.

[0210] 40℃ / 75%RH-closed means that it is placed in a sealed environment at 40℃ and 75% humidity.

[0211] 40℃ / 75%RH-open means to leave it open at 40℃ and 75% humidity.

[0212] 60℃-closed means placed in a sealed container at 60℃.

[0213] 40℃ / 75%RH-closed-2wks means that the product is stored in a sealed container for 2 weeks at 40℃ and 75% humidity.

[0214] 40℃ / 75%RH-open-2wks means to leave it open for 2 weeks at 40℃ and 75% humidity.

[0215] 60℃-closed-2wks means that it is placed in a sealed container at 60℃ for 2 weeks.

[0216] Initial refers to the initial state.

[0217] SGF stands for Simulated Gastric Juice.

[0218] FaSSIF refers to simulated intestinal fluid during fasting.

[0219] FeSSIF refers to intestinal fluid simulation during feeding.

[0220] 1d means 1 day; 3d means 3 days.

[0221] Preparation Example 1: Preparation of Compound I

[0222] The reaction formula for the synthesis of compound I and intermediate b

[0223]

[0224]

[0225] Preparation of intermediate b

[0226] In a 250 mL single-necked flask, dichloromethane (50 mL), 5-methoxy-7-methyl-1H-indole (3 g), Boc anhydride (5.68 g), 4-dimethylaminopyridine (227 mg), and triethylamine (2.26 g) were added sequentially, and the reaction was carried out at room temperature for 16 hours. After the reaction was completed, the reaction solution was quenched with saturated ammonium chloride solution (5 mL), extracted three times with dichloromethane (20 mL), the combined organic phases were washed with water (5 mL), dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated, and the residue was purified by column chromatography (petroleum ether:ethyl acetate = 10:1) to give intermediate a (4.6 g, yield: 94%). MS m / z (ESI): 262.0 [M+1].

[0227] In a 250 mL single-necked flask, dichloromethane (80 mL), N-methylformamide (3.8 g), and oxalyl chloride (3.6 g) were added sequentially, and the reaction was stirred at room temperature for 3 hours. The reaction temperature was then lowered to -14 °C, and intermediate a (2.5 g) was added. The reaction system was allowed to warm naturally to room temperature and stirred at room temperature for 1 hour. After the reaction was complete, the reaction mixture was poured into ice water (100 mL), extracted three times with dichloromethane (100 mL), and the combined organic phases were washed twice with water (10 mL). The mixture was dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated, and the residue was purified by column chromatography (petroleum ether:ethyl acetate = 20:1) to obtain intermediate b (1.3 g, yield: 47%).

[0228] MS m / z(ESI):290.0[M+1]. 1 H NMR (400MHz, CDCl3) δ 10.65 (s, 1H), 7.65 (d, J = 3.4Hz, 1H), 7.49 (d, J = 3.4Hz, 1H), 6.76 (s, 1H), 3.98 (s, 3H), 2.70 (s, 3H), 1.65 (s, 9H).

[0229] Preparation of Compound I

[0230] first step:

[0231] In a 3L three-necked flask, tetrahydrofuran (150mL) and 4-bromobenzonitrile (50g) were added sequentially. Under nitrogen protection, isopropyl magnesium chloride-lithium chloride complex (1.3M, 210mL) was slowly added to the reaction system, and the reaction was carried out at room temperature for 2 hours. Then, anhydrous tetrahydrofuran (500mL) was added to dilute the reaction system and the temperature was lowered to -5°C. 4-Methoxypyridine (25mL) was added, followed by the slow dropwise addition of benzyl chloroformate (35mL) (maintaining the system temperature below 0°C). After the addition was complete, the reaction was stirred at 0°C for 2 hours, then the temperature was raised to room temperature and the reaction was continued at room temperature for 16 hours. After the reaction was completed, 6M hydrochloric acid (150 mL) was added and stirred for half an hour. The mixture was then diluted with water (1000 mL) and extracted twice with ethyl acetate (500 mL). The combined organic phases were washed with saturated brine (50 mL), dried with anhydrous sodium sulfate, and filtered. The crude product obtained after concentration of the filtrate was purified by column chromatography (petroleum ether: ethyl acetate = 3:1-1:1) to give compound 1 (23 g, yield: 23%).

[0232] MS m / z(ESI):333.0[M+1].

[0233] Step Two:

[0234] 46g of compound 1 (28g), zinc powder (55g), and acetic acid (200mL) obtained from the first step of batch preparation were taken out and added sequentially to a 500mL single-necked flask. The reaction was heated to 100℃ and stirred at this temperature for 16 hours. After the reaction was completed, the mixture was filtered, and the filtrate was diluted with water (500mL). The filtrate was extracted with ethyl acetate (500mL), and the organic phase was washed twice with saturated sodium bicarbonate aqueous solution (500mL) and once with saturated brine (100mL). The mixture was dried with anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure to obtain compound 2 (26g, yield: 73%).

[0235] MS m / z(ESI): 334.8 [M+1].

[0236] Step 3:

[0237] In a 500 mL single-necked flask, tetrahydrofuran (100 mL), ethanol (100 mL), and compound 2 (26 g) were added sequentially, followed by the addition of sodium borohydride (2 g) in portions. The reaction was carried out at room temperature for 2 hours. After the reaction was complete, the system was cooled to 0 °C, and saturated ammonium chloride aqueous solution (30 mL) was added until the temperature no longer rose. The mixture was diluted with water (500 mL) and extracted twice with ethyl acetate (200 mL). The combined organic phases were washed with saturated brine (500 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure to give compound 3 (25 g, yield: 76%).

[0238] MS m / z(ESI): 336.9 [M+1].

[0239] Step 4:

[0240] Dichloromethane (200 mL) was added to a 500 mL single-necked flask, followed by the sequential addition of compound 3 (25 g), imidazole (6.6 g), and tert-butyldiphenylchlorosilane (25 g). The mixture was reacted at room temperature for 2 hours. After the reaction was complete, the reaction solution was washed with water (500 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by column chromatography (petroleum ether:ethyl acetate = 10:1) to give compound 4 (5.7 g, yield: 13%, Rf = 0.55; cis isomer Rf = 0.50).

[0241] MS m / z(ESI):597.0[M+23].

[0242] Step 5:

[0243] In a 250 mL single-necked flask, compound 4 (5 g) and tetrabutylammonium fluoride tetrahydrofuran solution (1 M, 30 mL) were added sequentially, and the reaction was carried out at room temperature for 2 hours. After the reaction was completed, the mixture was diluted with water (100 mL), extracted three times with ethyl acetate (50 mL), and the combined organic phases were washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by column chromatography (petroleum ether:ethyl acetate = 3:1-0:1) to obtain a racemic mixture. This racemic mixture was chirally separated by SFC (Apparatus: SFC Thar prep 80; Column: CHIRALPAK AD-H, 250 mm × 20 mm, 5 μm; Modifier: 35% methanol (0.2% ammonia); Column temperature: 40 °C; Column pressure: 60 bar; Wavelength: 214 / 254 nm; Flow rate: 40 g / min; Rt = 4.78 min) to obtain compound 5 (1.2 g, yield: 41%).

[0244] MS m / z(ESI):358.8[M+23].

[0245] Step 6:

[0246] To a solution of compound 5 (1200 mg) in N,N-dimethylformamide (10 mL), imidazole (486 mg) and tert-butyldimethylchlorosilane (593 mg) were added, and the reaction was stirred at room temperature for 2 hours. After the reaction was complete, the reaction mixture was diluted with water (100 mL), extracted with ethyl acetate (50 mL), washed once with saturated brine (50 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was directly concentrated to give compound 6 (600 mg, yield: 90%).

[0247] MS m / z(ESI):472.8[M+23].

[0248] Step 7:

[0249] At room temperature, 1.2 g of compound 6 (700 mg) from the second batch of step 6 was added to dichloromethane (10 mL). Under nitrogen protection and at -78°C, cyclopropylformaldehyde (110 mg) and trimethylsilyl trifluoromethanesulfonate (35 mg) were added to the reaction solution. The reaction system was maintained at -78°C and stirred for one hour. Then, triethylsilane (180 mg) was added, and the reaction was allowed to rise naturally to room temperature and stirred for another 16 hours. After the reaction was completed, the reaction solution was quenched with saturated sodium bicarbonate aqueous solution (20 mL), diluted with water (10 mL), extracted with dichloromethane (10 mL), washed once with water (10 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated, and the residue was purified by column chromatography (petroleum ether: ethyl acetate = 3:1) to obtain compound 7 (400 mg, yield: 46%).

[0250] MS m / z(ESI): 390.9 [M+1].

[0251] Step 8:

[0252] Compound 7 (400 mg), isopropanol (2 mL), water (3 mL), and sodium hydroxide (400 mg) were added sequentially to a 50 mL single-necked flask. The reaction mixture was heated to 100 °C and stirred at this temperature for 16 hours. After the reaction was completed, the pH of the reaction solution was adjusted to 5-6 by adding dilute hydrochloric acid (1 M) in an ice bath, diluted with water (5 mL), extracted with ethyl acetate (5 mL), washed once with saturated brine (5 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated at 45 °C to give compound 8 (200 mg, yield: 33%).

[0253] MS m / z(ESI):431.8[M+23].

[0254] Step 9:

[0255] Potassium carbonate (135 mg) and iodomethane (140 mg) were added to a solution of compound 8 (200 mg) in acetonitrile (5 mL). The reaction mixture was heated to 50 °C and stirred at that temperature for 16 hours. After the reaction was completed, the reaction mixture was directly concentrated, and the residue was purified by column chromatography (petroleum ether: ethyl acetate = 3:1) to give compound 9 (180 mg, yield: 40%).

[0256] MS m / z(ESI):445.8[M+23].

[0257] Step 10:

[0258] Palladium / carbon (50 mg) was added to a tetrahydrofuran (3 mL) solution of compound 9 (180 mg), and the reaction solution was subjected to catalytic hydrogenation at room temperature under a hydrogen atmosphere for 2 hours. After the reaction was completed, the reaction solution was filtered, and the filtrate was directly concentrated to give compound 10 (120 mg, yield: 54%).

[0259] MS m / z(ESI):290.0[M+1].

[0260] Step 11:

[0261] Compound 10 (120 mg) was added to a solution of intermediate b (119 mg) in 1,2-dichloroethane (5 mL), and the reaction was stirred at room temperature for 8 hours. Then, sodium borohydride acetate (261 mg) was added, and the mixture was stirred at room temperature for another 16 hours. After the reaction was complete, the reaction solution was directly concentrated, and the residue was purified by column chromatography (dichloromethane:methanol = 20:1) to give compound 11 (200 mg, yield: 26%).

[0262] MS m / z(ESI): 562.8 [M+1].

[0263] Step Twelve

[0264] Methanol (2 mL), water (2 mL), compound 11 (200 mg), and sodium hydroxide (150 mg) were added sequentially to a 50 mL single-necked flask. The reaction mixture was heated to 75 °C and stirred at this temperature for 3 hours. After the reaction was completed, the pH of the reaction solution was adjusted to 7 by adding dilute hydrochloric acid (1 M) in an ice bath, and then concentrated under reduced pressure and purified by Prep-HPLC (column: Gemini-C18, 150 x 21.2 mm, 5 μm; mobile phase: acetonitrile-water (0.1% formic acid); gradient: 20-40%) to give compound I (30.6 mg, yield: 18%; containing 0.5 equivalents of formic acid).

[0265] MS m / z(ESI):448.9[M+1].

[0266] 1H NMR (400MHz, CD3OD): δ8.18(d,J=7.7Hz,2H),7.69(d,J=7.7Hz,2H),7.32(s,1H),6 .76(s,1H),6.34(s,1H),4.88-4.61(m,1H),4.44-4.07(m,2H),3.95-3.81(m,1H),3 .75(s,3H),3.63-3.47(m,1H),3.46-3.33(m,3H),2.50(s,3H),2.35-2.14(m,2H), 2.13-1.94(m,2H),1.23-1.04(m,1H),0.58(d,J=7.2Hz,2H),0.28(d,J=3.8Hz,2H).

[0267] Unless otherwise stated, all compounds of formula I mentioned below are compounds of formula I prepared by the above method or by repeating the above method.

[0268] Example 1: Preparation method of monohydrochloride salt of compound I

[0269]

[0270] After multiple batches of preparation of the compound of formula I, 400 mg of the compound was dissolved in 8 mL of isopropanol by heating at 50 °C. Then, 460 μL of isopropanol hydrochloride solution (2 mol / L) was slowly added dropwise, and the mixture was stirred for half an hour. Next, 8 mL of n-heptane was added, and the mixture was stirred for another 2 hours. The mixture was filtered, and the filter cake was dried under reduced pressure at 50 °C to obtain 390 mg of the monohydrochloride salt I-1 of the compound of formula I, with a yield of 90%.

[0271] Take 50 mg of compound I monohydrochloride I-1 into a 4 mL sample vial and add 0.5 mL of methanol to dissolve the compound. Then, place the sample open into a 40 mL sample vial containing 5 mL of ethyl acetate, keeping the 40 mL sample vial sealed. Allow it to stand to allow the two solvents to diffuse slowly, yielding a single crystal. The single crystal structure diagram is shown below. Figure 24 The single crystal was tested, and the single crystal data of compound I monohydrochloride I-1 were obtained as follows:

[0272]

[0273]

[0274] Example 2: Preparation method of phosphate of compound I

[0275] Compound I was prepared in multiple batches. 440 mg of compound I was added to 5 mL of acetone, heated to 40 °C, and sonicated to dissolve. Then, 460 μL of a 2 mol / L phosphate methanol solution was slowly added dropwise. A viscous solid was observed; another 5 mL of acetone was added, and the mixture was stirred at room temperature for 4 hours. The mixture was filtered, washed, and the filter cake was dried under reduced pressure at 50 °C to obtain 462 mg of phosphate of compound I, with a yield of 87%.

[0276] Example 3: Preparation method of maleate salt of compound I

[0277] Compound I was prepared in multiple batches. 400 mg of compound I was added to 15 mL of ethyl acetate, heated to 50 °C, and the compound was dissolved. Then, 109 mg of maleic acid powder was added, and the mixture was stirred at room temperature for 2–3 hours. The mixture was filtered, and the filter cake was dried under reduced pressure at 50 °C to obtain 470 mg of maleate of compound I, with a yield of 91%.

[0278] Example 4: Stability test of compound I monohydrochloride, phosphate, and maleate.

[0279] The stability of the compound of formula I, the monohydrochloride of the compound of formula I in Example 1, the phosphate of the compound of formula I in Example 2, and the maleate of the compound of formula I in Example 3 were investigated.

[0280] Stability testing conditions: 40℃ / 75%RH-closed, 40℃ / 75%RH-open, 60℃-closed; Stability testing content: changes in related substances and crystal forms.

[0281] Related substances detection: Weigh approximately 6–7 mg of sample into 10 mL volumetric flasks, dissolve and dilute to the mark with 50% acetonitrile aqueous solution, and inject 10 μL. Chromatographic conditions are shown in Table 6.

[0282] The results of the stability tests of the compounds of Formula I and their hydrochloride, phosphate and maleate are shown in Table 7.

[0283] Table 6. Chromatographic conditions for testing compounds of formula I and their monohydrochloride, phosphate, and maleate salts.

[0284]

[0285]

[0286] Table 7. Results of stability studies for compounds of Formula I and their monohydrochloride, phosphate, and maleate salts.

[0287]

[0288] As can be seen from the results in Table 7, the compounds of Formula I and their monohydrochloride, phosphate and maleate are very stable, especially the monohydrochloride of the compounds of Formula I, which has excellent stability.

[0289] Example 5: Solubility test of Formula I compound and its hydrochloride and phosphate salts

[0290] The solubility of compound I, compound I monohydrochloride, and compound I phosphate in water, SGF, FaSSIF, and FeSSIF was investigated at 37°C.

[0291] Experimental method: Weigh 30 mg (in water) or 15 mg of sample into a 4 mL vial, add 3 mL of the test medium (water, SGF, FaSSIF, FeSSIF), and stir continuously at 37℃. Take 0.5 mL samples at 1 h and 24 h respectively, centrifuge at 12000 rpm for 10 min, and dilute the supernatant with 50% acetonitrile aqueous solution to an appropriate factor before determining its concentration. The chromatographic conditions for solubility testing are shown in Table 8.

[0292] Reference Standard and Linearity: Weigh 10 mg of compound I into a 50 mL volumetric flask, dissolve in 50% acetonitrile aqueous solution, and dilute to the mark. Prepare two parallel aliquots. Take the reference standard of compound I, dilute with 50% acetonitrile aqueous solution to 100 μg / mL, 50 μg / mL, and 10 μg / mL, inject 5 μL, and plot a standard curve.

[0293] The solubility test results of the compounds of Formula I and their hydrochlorides and phosphates are shown in Table 9.

[0294] Table 8. Chromatographic conditions for solubility testing of compound I.

[0295] Liquid phase equipment Agilent 1260 chromatographic column Waters Sunfire C18(4.6*150mm, 5μm) mobile phase A: 0.1% TFA aqueous solution; B: 0.1% TFA acetonitrile solution Isocratic elution A:B = 60:40 Detection wavelength (nm) 230 Injection volume (μL) 5 Sample concentration (mg / mL) 0.5 Column temperature (°C) 40 Flow rate (mL / min) 1.0 diluent 50% acetonitrile aqueous solution Running time (min) 30

[0296] Table 9. Solubility test results of Formula I compounds and their hydrochlorides and phosphates.

[0297]

[0298]

[0299] As can be seen from Table 9, the solubility of compound I in water increases after it forms a salt.

[0300] control compound

[0301]

[0302] Methanol (3 mL), water (1 mL), intermediate 1 (160 mg), and sodium hydroxide (230 mg) were added to a 50 mL single-necked flask. The reaction was carried out at room temperature for 16 hours. After the reaction was completed, water (10 mL) was added to dilute the mixture, and the pH was adjusted to 7-8 with dilute hydrochloric acid solution (1 M). The solvent was removed under reduced pressure (water bath: 45 °C). The residue was purified by preparative high-performance liquid chromatography (HPLC) (column: Gemini-C18, 150 x 21.2 mm, 5 μm; mobile phase: acetonitrile-water (0.1% formic acid); gradient: 15-30%) to obtain the control compound (29 mg, yield: 24%). MS m / z (ESI): 423.1 [M+1]. 1 H NMR (400MHz, DMSO-d6) δ8.17(d,J=8.4Hz,2H),7.67(d,J=8.4Hz,2H),7.33(t,J= 2.8Hz,1H),6.78(s,1H),6.35(s,1H),4.82–4.67(m,1H),4.40–4.17(m,2H),3.90 –3.81(m,1H),3.77(s,3H),3.62(q,J=6.8Hz,2H),3.57–3.50(m,1H),3.45–3.35( m,1H),2.52(s,3H),2.32–2.22(m,2H),2.14–1.96(m,2H),1.32(t,J=6.8Hz,3H).

[0303] Biological Example 1

[0304] 1. Optical Surface Plasmon Resonance (SPR) Bonding Test

[0305] The SPR experiment was conducted at 25°C using PBS buffer supplemented with 0.05% (v / v) P20 and 5% DMSO as the run buffer. The analytical instrument used was a GE Healthcare Biacore 8K. The CM7 chip (GE Healthcare) was activated for 420 s with 400 mM EDC and 100 mM NHS at a flow rate of 30 μL / min. Complement B factor was diluted to 50 μg / mL with 10 mM sodium acetate (pH 4.0) and then coupled at a flow rate of 10 μL / min for 1200 s to covalently immobilize complement B factor onto the detection chip (protein immobilization level: 25000 RU). The detection chip was then blocked with 1 M ethanolamine hydrochloride at a flow rate of 10 μL / min for 300 s. The analyte concentration was 500 μM, with a binding time of 120 s and a dissociation time of 300 s. The data analysis was performed using a 1:1 binding model (Biacore Insight Evaluation Software, Version 2.0.15.12933).

[0306] Experimental results:

[0307] The experimental results are shown in Table 10. At a concentration of 500 μM, compound I exhibited a more significant binding affinity to the target protein, which was significantly better than that of the control compound, indicating that compound I has a better binding affinity to the target protein.

[0308] Table 10

[0309] sample Rmax(RU) Formula I compound 274.2 control compound 33.7

[0310] 2. TR-FRET bonding strength test

[0311] Competitive binding assays using Cy5-labeled small molecule inhibitors as probes were employed to screen compounds for their inhibitory activity against human complement factor B. Complement factor B and EZ-Link... TM The Sulfo-NHS-LC-LC-Biotin mixture was incubated on ice at a 1:2 ratio for 1 hour, followed by termination with 1M Tris (pH 7.5). Then, 2 mL of Zeba solution was added. TMBiotin-labeled complement factor B was obtained by purification twice using a desalt spin column (EZ-Link™ Sulfo-NHS-LC-Biotin instructions). For the experiment, 10 nM biotin-labeled complement factor B was pre-incubated with different concentrations of the compound in buffer at room temperature for 1 hour. Cy5 fluorescently labeled probes and europium chelate-labeled streptavidin (petroleum ether rkin Elmer, #AD0060) were added at final concentrations of 75 nM and 5 nM to initiate the reaction. Kinetic readings were performed on a microplate reader (337 nm excitation, 665 nm emission, 70 μs time-gated), and time-dependent fluorescence energy transfer (TR-FRET) data were recorded to determine the IC50. 50 .

[0312] 3. Detection of C3 hydrolysis activity in the complement system

[0313] The test compound was initially tested at a concentration of 10 μM, then diluted 3-fold, with 7 concentration points, and detected in a single well. In a 96-well plate, the test compound was diluted to a final concentration of 1000-fold with DMSO, and then analyzed using Diluent (…). The COMPLEMENT SYSTEM ALTERNATIVE PATHWAY AP330 was diluted to a final concentration of 5. 30 μL was transferred to a 96-well plate, and 120 μL of spare serum was added. The plate was incubated at room temperature for 15 minutes. 30 μL of 5‰ DMSO and 120 μL of spare serum were added to the positive control wells, and 30 μL of 5‰ DMSO and 120 μL of Diluent were added to the negative control wells. (3) 100 μL was added to the reaction plate and incubated at 37°C for 60 minutes. The liquid in the wells was discarded, and each well was washed three times with 300 μL of washing buffer. 100 μL of Conjugate (…) was added to each well. COMPLEMENT SYSTEM ALTERNATIVE PATHWAY AP330), incubate at room temperature for 30 minutes. Discard the liquid in the wells and wash each well three times with 300 μL of washing buffer. Then add 100 μL of substrate to each well and incubate at room temperature for 30 minutes. Detect using a microplate reader (PerkinElmer, EnSight) and read the OD405 value.

[0314] 4. Complement hemolytic activity assay

[0315] The hemolysis assay was described in reference Xuan Yuan et al., Haematologica, (2017) 102:466-475. Prior to the experiment, the optimal concentration of normal human serum (NHS) required to achieve 100% lysis of rabbit erythrocytes (REs) was determined by titration. In this experiment, NHS was diluted in GVB0 buffer containing 10 mM Mg-EGTA (0.1% gelatin, 5 mM Veronal, 145 mM NaCl, 0.025% NaN3, pH 7.3, Complement technology) and incubated with various concentration gradients of test compounds at 37°C for 15 min. Freshly resuspended REs (from healthy Japanese White rabbits) in GVB0 buffer containing 10 mM Mg-EGTA were added to a final concentration of 1 × 10⁸ cells / mL and incubated at 37°C for 30 min. The positive control group (100% lysis) consisted of GVB0 buffer containing 10 mM Mg-EGTA with NHS and RE but without the test compound; the negative control group (0% lysis) consisted of GVB0 buffer containing 10 mM Mg-EGTA with inactivated NHS (heated at 56°C for 30 min or 65°C for 5 min) and RE but without the test compound. Samples were centrifuged at 2000 g for 5 min, and the supernatant was collected. The absorbance at 415 nm (A415) was detected using a microplate reader (Molecular Devices, SpectraMax i3X). IC50 50 The value was calculated from the percentage of hemolysis as a function of the concentration of the test compound using nonlinear regression.

[0316] Experimental results:

[0317] The experimental results are shown in Table 11. The compound of formula I showed significantly better inhibitory activity against complement factor B in human serum than the control compound, indicating that the compound of the present invention can effectively inhibit the activity of complement factor B in human serum and prevent hemolysis caused by its attack on rabbit erythrocytes.

[0318] Table 11

[0319] sample <![CDATA[Hemolytic IC 50 (nM)]]> Formula I compound 87.9 control compound 379.4

[0320] 5. Liver microsomal stability test

[0321] (1) Preparation of buffer solution

[0322] Take a 0.1M dipotassium hydrogen phosphate distilled aqueous solution (containing 1mM ethylenediaminetetraacetic acid), and then adjust the pH to 7.4 with a 0.1M dipotassium hydrogen phosphate distilled aqueous solution (containing 1mM ethylenediaminetetraacetic acid).

[0323] (2) Microparticle source and working solution preparation

[0324] Microsome source:

[0325] Rats: SD Rat Liver Microsomes, Cat. No.: LM-DS-02M, RILD Liver Disease Research (Shanghai) Co., Ltd.

[0326] Monkey: Cynomolgus Monkey Liver Microsomes, Cat. No.: LM-SXH-02M, RILD Liver Disease Research (Shanghai) Co., Ltd.

[0327] Human: Pooled Human Liver Microsomes (Mongolian), Cat. No.: LM-R-02M, RILD Liver Disease Research (Shanghai) Co., Ltd.

[0328] Preparation of working fluid

[0329] Prepare 10 mM solutions of both the control and test compounds using DMSO. Then, add 10 μL of each solution to 190 μL of acetonitrile to prepare a 0.5 mM stock solution. Take 1.5 μL of the 0.5 mM stock solution, add 18.75 μM of 20 mg / mL liver microsomes, and 479.75 μL of buffer solution. (The actual prepared volume may be adjusted according to usage requirements.)

[0330] (3) Experimental procedure

[0331] Prepare 10 mg / mL of reduced coenzyme II (NADPH) using buffer. Place a 96-well plate on ice, and set up corresponding wells for different time points for each compound (0, 10, 30, 60, 90 minutes, Non-NADPH). Add 30 μL of working solution to each well. For the 0 min well, first add 155 μL of ice-cold acetonitrile solution (internal standard concentration of 1 μM), mix with a pipette, and then add 15 μL of NADPH (10 mg / mL). Before starting the reaction, pre-incubate the 96-well plate on a microplate shaker (37°C) for 5 minutes, and then add 15 μL of NADPH (10 mg / mL) to each well to start the metabolic reaction. After 10, 30, 60, and 90 minutes of reaction, add 155 μL of ice-cold acetonitrile solution (internal standard concentration of 1 μM) to the corresponding well to terminate the reaction. After 90 minutes, the non-NADPH system was terminated by adding 155 μL of ice-cold acetonitrile solution (internal standard concentration of 1 μM). After the reaction, the 96-well plate was shaken for 10 minutes using a microplate shaker (600 rpm), then centrifuged at 4℃ and 4000g for 15 minutes. 50 μL of the supernatant was added to a new 2 mL 96-well plate, followed by 300 μL of deionized water. Analysis was performed using an AB SCIEX ExionLC-Triple Quad 5500 high-performance liquid chromatography-mass spectrometry system with Analyst 1.6.3 software. The results are shown in Table 12.

[0332] Table 12

[0333]

[0334] Experimental results: Data show that compound I has significant liver microsomal stability.

[0335] 6. Single-dose gavage administration pharmacokinetic experiment in rats

[0336] Experimental methods:

[0337] Six- to nine-week-old male Wistar Han rats (Shanghai Xipu-Bikai Experimental Animal Co., Ltd.) were used. After overnight fasting, three rats per group were administered the drugs via gavage. The control compound and compound I were each administered 3 mg / kg, at a volume of 10 mL / kg. Blood was collected via the jugular vein, 0.2 mL at each time point, anticoagulated with EDTA-K2, and immediately centrifuged at 4000 rpm for 5 min at 4°C. The supernatant was collected and stored at -80°C until analysis. Blood collection time points were: before administration, 5 min, 15 min, 30 min, 1 h, 2 h, 4 h, 7 h, and 24 h. Animals were monitored after administration, and euthanized after blood collection at all time points. Plasma samples were analyzed using LC-MS / MS, and kinetic parameters (Tmax, Cmax, T1 / 2, AUC) were calculated using WinNonlin software.

[0338] Experimental results:

[0339] The test results are shown in Table 13.

[0340] Table 13

[0341]

[0342] 7. Single-dose gavage administration pharmacokinetic / probabilistic (PK / PD) experiment in cy

[0343] Experimental methods:

[0344] Cynomolgus monkeys were used, with 3 monkeys in each group receiving 3 mpk of the control compound, 30 mpk of the control compound, 3 mpk of compound I, and 30 mpk of compound I via gavage. Blood samples were collected at different time points for drug concentration analysis and complement activity detection. Plasma compound concentrations were determined by LC-MS / MS, and serum complement activity was detected using the wieslab assay kit (Svar Life Science AB, COMPL AP330 RUO), including Normal Human Serum (Complement Technology, NHS).

[0345] Experimental results:

[0346] Within the tested concentration and time range, at the same dose, the average blood concentration of compound I was significantly higher than that of the control compound. Blood concentration curves in cynomolgus monkeys are shown below. Figure 21 Inhibition of AP activity in cynomolgus monkey serum was observed. Figure 22 . Figure 22 The results show that compound I can effectively inhibit AP activity in cynomolgus monkey serum.

[0347] 8. Streptococcus-induced rat model of rheumatoid arthritis (RA)

[0348] Experimental methods:

[0349] The experiment used 6-9 week old female Lewis rats (Beijing Vital River Pharmaceutical Co., Ltd.), with 6 rats in each group. On day 1, rats were intraperitoneally injected with a peptidoglycan complex containing the cell walls of streptococci and several other bacteria (2-3 mg per rat). The control compound (15 mpk) and compound I (15 mpk) were administered by gavage daily for 25 days. Arthritis scores were assessed at different time points. The scoring criteria were as follows: Rats were scored from 0 to 4 points based on the severity of the lesions (redness and swelling), with a maximum score of 4 points for each limb and a maximum total score of 16 points for all four limbs per animal. The scoring criteria were as follows: 0 points, no redness or swelling; 1 point, redness and swelling of 1-2 interphalangeal joints; 2 points, redness and swelling of 3-4 interphalangeal joints; 3 points, redness and swelling of more than 4 interphalangeal joints; 4 points, severe redness and swelling from the toes or fingers to the ankle or wrist joints.

[0350] Experimental results:

[0351] The experimental results are shown in Figure 23 Data showed that compound I could improve arthritis scores, and its effect was significantly better than that of the control compound, proving that compound I could more effectively improve streptococcal-induced rheumatoid arthritis in rats.

[0352] Example 6: Preparation method of monohydrochloride crystal form A of compound I

[0353] Take 400 mg of compound I and add 8 mL of isopropanol, heating to dissolve at 50 °C. Then, slowly add 460 μL of isopropanol hydrochloride solution (concentration 2 mol / L), stirring for half an hour, then add 8 mL of n-heptane, and continue stirring for 2 hours. Filter, and dry the filter cake under reduced pressure at 50 °C to obtain 390 mg of compound I monohydrochloride crystal form A, with a yield of 90%.

[0354] Crystal form A was characterized by XRPD, DSC, TGA and PLM.

[0355] Crystal form A is an anhydrous compound. The positions and intensities of the characteristic XRPD peaks are shown in Table 1, and the XRPD spectrum is shown below. Figure 1 As shown.

[0356] The DSC display showed that the first endothermic peak appeared near the peak temperature of 192.73℃, and the first exothermic peak appeared near the peak temperature of 201.78℃. Figure 2 As shown.

[0357] TGA showed a weight loss of approximately 1.41% in the temperature range of 90°C to 180°C, such as Figure 3 As shown.

[0358] The PLM image shows that the sample is an irregularly shaped crystal with a diameter of less than 20 μm, such as Figure 4 As shown.

[0359] The XRPD pattern of crystal form A, expressed in 2θ angles, shows the 2θ angles and relative intensities of the diffraction peaks in Table A. The error range for the 2θ angles is ±0.20°.

[0360] Table A shows the XRPD analytical data for crystal form A.

[0361]

[0362]

[0363] Example 7: Preparation method of monohydrochloride crystal form B of compound formula I

[0364] Crystal form A, when left exposed to accelerated conditions (40°C / 75% RH) for 72 hours, will transform into crystal form B. Crystal form B is a monohydrate of the monohydrochloride salt of compound formula I.

[0365] Crystal form B was characterized by XRPD, DSC, TGA and PLM.

[0366] The positions and intensities of the characteristic peaks of XRPD are shown in Table 2, and the XRPD spectrum is shown in... Figure 5 As shown.

[0367] DSC analysis showed that the first endothermic peak appeared near the peak temperature of 85.87℃, the second endothermic peak appeared near the peak temperature of 197.54℃, and the first exothermic peak appeared near the peak temperature of 205.68℃. Figure 6 As shown.

[0368] TGA showed approximately 3.42% weight loss in the 21.49℃ to 120℃ range and approximately 0.49% weight loss in the 179.88℃ to 207.94℃ range. Figure 7 As shown.

[0369] The PLM image shows that the sample is an irregularly shaped crystal with a diameter of less than 20 μm, such as Figure 8 As shown.

[0370] The XRPD pattern of crystal form B, expressed as a 2θ angle in the X-ray powder diffraction pattern, shows the 2θ angles and relative intensities of the diffraction peaks as shown in Table B, where the error range of the 2θ angle is ±0.20°.

[0371] Table B shows the XRPD analysis data for crystal form B.

[0372] Peak number 2θ[°] Relative strength % Peak number 2θ[°] Relative strength % 1 9.48 32.4 13 23.40 8.2 2 10.78 22.6 14 24.10 13.3 3 12.08 7.1 15 25.41 7.8 4 14.68 14.5 16 26.22 9 5 15.44 27.8 17 28.16 14.4 6 18.10 51.2 18 29.18 8 7 19.18 20.7 19 29.58 5.5 8 19.80 49.9 20 30.13 5.2 9 20.60 17.9 21 30.92 24.4 10 21.34 12.4 22 33.02 4.6 11 22.10 100 23 35.50 8.7 12 22.94 9.3 24 41.28 4.8

[0373] Example 8: Preparation method of monohydrochloride crystal form C of compound I

[0374] Compound I (3.15 g) was added to a three-necked flask, followed by 12.6 mL of methanol and stirring until completely dissolved. A methanol solution of 1.8 N HCl (3.05 g) was added dropwise at room temperature, and the mixture was stirred for 10 minutes before filtration. The filtrate was added to a three-necked flask, and 78.75 mL of methyl tert-butyl ether was added dropwise at room temperature. The mixture was stirred for 2 hours, filtered, and the filter cake was dried to give crystal form C (3.06 g), with a yield of 90%.

[0375] Crystal form C is an anhydrous form, and it was characterized by XRPD, DSC, TGA and PLM.

[0376] The positions and intensities of the characteristic peaks of XRPD are shown in Table 3, and the XRPD spectrum is shown in... Figure 9 As shown.

[0377] The DSC display showed the first endothermic peak near the peak temperature of 209.93℃ and the first exothermic peak near the peak temperature of 215.80℃. Figure 10 As shown.

[0378] TGA showed approximately 0.29% weight loss in the range of 21.62℃ to 120℃ and approximately 0.52% weight loss in the range of 173.94℃ to 216.60℃. Figure 11 As shown.

[0379] The PLM image shows that the sample is an irregularly shaped crystal with a diameter of less than 20 μm, such as Figure 12 As shown.

[0380] The XRPD pattern of crystal form C, expressed as a 2θ angle in its X-ray powder diffraction pattern, shows the 2θ angles and relative intensities of the diffraction peaks as shown in Table C:

[0381] XRPD analysis data for crystal form C (Table C)

[0382] Peak number 2θ[°] Relative strength % Peak number 2θ[°] Relative strength % 1 10.00 15.7 14 21.98 69.7 2 10.52 12.7 15 22.94 36.2 3 11.96 21 16 24.80 17.1 4 12.48 12.7 17 25.92 40.4 5 13.62 13.2 18 27.46 22.7 6 14.74 100 19 29.06 27.8 7 15.58 10.9 20 30.30 16.3 8 17.80 70.2 21 31.54 6.1 9 18.58 17.7 22 32.10 13.2 10 18.90 14.9 23 33.48 29.3 11 19.58 43.8 24 33.94 7.5 12 20.08 88.8 25 36.16 6.8 13 21.06 20 26 42.06 9.5

[0383] Example 9: Preparation method of monohydrochloride crystal form D of compound I

[0384] Take 400 mg of compound I monohydrochloride, add 8 mL of dichloromethane, and stir at room temperature for 24 hours. Separate the resulting suspension by centrifugation, and dry the solid under reduced pressure at 40 °C. The solid is crystal form D.

[0385] The crystal form D is a dichloromethane solvate of the monohydrochloride salt of compound I (or called monodichloromethane solvate).

[0386] Crystal form D was characterized by XRPD, DSC, TGA and PLM.

[0387] The positions and intensities of the characteristic peaks of XRPD are shown in Table 4, and the XRPD spectrum is shown in... Figure 13 As shown.

[0388] The DSC display showed the first exothermic peak occurring near the peak temperature of 196.53℃. Figure 14 As shown.

[0389] TGA showed a weight loss of approximately 6.31% in the temperature range of 22.07°C to 120°C. Figure 15 As shown.

[0390] The PLM image shows that the sample is an irregularly shaped crystal with a diameter of less than 10 μm, such as Figure 16 As shown.

[0391] The XRPD pattern of crystal form D, expressed as a 2θ angle in its X-ray powder diffraction pattern, shows the 2θ angles and relative intensities of the diffraction peaks as shown in Table D:

[0392] XRPD analytical data for crystal form D in Table D

[0393] Peak number 2θ[°] Relative strength % Peak number 2θ[°] Relative strength % 1 10.16 37.6 13 23.82 43.9 2 11.90 25.6 14 24.94 11.8 3 12.60 14.6 15 25.60 9.8 4 15.74 100 16 26.24 23.3 5 16.58 43.5 17 26.80 11.7 6 19.22 40.1 18 27.50 12.9 7 19.80 13.9 19 29.36 7 8 20.24 26.4 20 29.88 13.6 9 21.12 9.8 21 31.00 6.6 10 21.98 41.1 22 32.48 4.8 11 22.66 10.4 23 36.20 6.3 12 23.18 14.4 24 36.76 4.4

[0394] Example 10: Preparation method of monohydrochloride crystal form E of compound I

[0395] Take 400 mg of compound I monohydrochloride, add 8 mL of isopropanol, and stir at room temperature for 72 hours. Separate the resulting suspension by centrifugation, and dry the solid under reduced pressure at 40 °C. The solid is crystal form E.

[0396] The crystal form E is an isopropanol solvate of the monohydrochloride salt of compound I.

[0397] XRPD characterization of crystal form E was performed, and the XRPD spectrum is shown below. Figure 17 As shown.

[0398] The XRPD pattern of crystal form E, expressed as a 2θ angle in its X-ray powder diffraction pattern, shows the 2θ angles and relative intensities of the diffraction peaks as shown in Table E:

[0399] XRPD analysis data for crystal form E in Table E

[0400]

[0401]

[0402] Example 11: Stability study of monohydrochloride crystal form C of compound I

[0403] The monohydrochloride crystal form C of compound I was placed under the conditions of 40℃ / 75%RH-closed, 40℃ / 75%RH-open, and 60℃-open, and removed after 1 month to investigate the stability of its crystal form.

[0404] The chromatographic conditions are shown in Table 14.

[0405] Stability of the sample related substances detection method: Weigh approximately 6 mg of the sample into a clean 40 mL glass bottle, add 10 mL of 50% acetonitrile aqueous solution, sonicate to completely dissolve, and inject 10 μL for related substances testing, as shown in Table 15. The XRPD diagram is shown in Table 18, where HCl-salt Form 3-initial (hereinafter referred to as initial) represents the XRPD diagram of crystal form C prepared in Example 8.

[0406] Table 14 Chromatographic Conditions Test

[0407]

[0408] Table 15 shows the stability test results of the monohydrochloride crystal form C of compound I (1M).

[0409]

[0410] Note: 1M represents 1 month of storage.

[0411] The data above shows that the monohydrochloride crystal form C of compound I exhibits good chemical stability after being stored at 40℃ / 75%RH and 60℃ for one month. Furthermore, from... Figure 18 It is evident that the crystal form of the monohydrochloride salt C of compound I remains unchanged.

[0412] Example 12: Effects of High Temperature and High Humidity on Crystal Form C

[0413] The monohydrochloride crystal form C of compound I was exposed to open conditions of 60°C, 80% RH, and 92.5% RH to evaluate whether significant changes in crystal form occurred. XRPD was measured at 1 day and 3 days. XRPD is as follows... Figure 19 , 20 As shown.

[0414] from Figure 19 , Figure 20 It can be seen that the monohydrochloride crystal form C of compound I did not change after being exposed to high temperature and high humidity conditions for 1 day and 3 days.

[0415] Example 13: Solubility test of compound I, mixture of crystal forms A and B, and crystal form C.

[0416] Weigh an appropriate amount of the test sample into a vial, add 3 mL of medium (water, SGF, FaSSIF, FeSSIF, etc.), and stir at 37℃. Take appropriate amounts of sample at 1 h and 24 h, centrifuge at 12000 rpm for 10 min, and dilute the supernatant with 50% acetonitrile aqueous solution to an appropriate factor before determining its concentration. The chromatographic conditions for solubility testing are shown in Table 16.

[0417] Reference Standard and Linearity: Weigh 10 mg of compound I into a 50 mL volumetric flask, dissolve in 50% acetonitrile aqueous solution, and dilute to the mark. Prepare two parallel aliquots. Take the reference standard, dilute with 50% acetonitrile aqueous solution to 100 μg / mL, 50 μg / mL, and 10 μg / mL, inject 5 μL, and plot a standard curve.

[0418] The test results are shown in Table 17.

[0419] Table 16 Chromatographic conditions for testing the solubility of compound I monohydrochloride salts

[0420]

[0421]

[0422] Table 17 Solubility results of compound I, mixture of crystal forms A and B, and crystal form C (37℃)

[0423]

[0424] Note: The pH values ​​at pH 6.8 and pH 7.4 were adjusted with sodium hydroxide solution, so the final pH values ​​are close to the initial values.

[0425] The experimental results above show that crystal form C has lower solubility in water compared to a mixture of crystal forms A and B.

[0426] The exemplary embodiments of the present invention have been described above. However, the scope of protection of this application is not limited to the exemplary embodiments described above. Any modifications, equivalent substitutions, improvements, etc., made by those skilled in the art within the spirit and principles of the present invention should be included within the scope of protection defined by the claims of this application.

Claims

1. A pharmaceutically acceptable salt of a compound of formula I: （I） The pharmaceutically acceptable salt is selected from its hydrochloride, sulfate, phosphate, methanesulfonate, p-toluenesulfonate, fumarate, maleate, citrate, L-tartrate, and oxalate.

2. A pharmaceutically acceptable salt of the compound of formula I according to claim 1, characterized in that, The pharmaceutically acceptable salt is the salt formed by the compound of formula I and hydrochloric acid.

3. A pharmaceutically acceptable salt of the compound of formula I according to claim 1, characterized in that, The pharmaceutically acceptable salt is the monohydrochloride salt formed by the compound of formula I and hydrochloric acid.

4. A method for preparing a pharmaceutically acceptable salt of the compound of formula I according to claim 1, characterized in that, The preparation method includes reacting a compound of formula I with a corresponding acid to prepare a pharmaceutically acceptable salt of the compound of formula I.

5. The preparation method according to claim 4, characterized in that, The preparation method includes reacting a compound of formula I with a corresponding acid in a solvent to prepare a pharmaceutically acceptable salt of the compound of formula I; The solvent is selected from alcohols, ketones, esters, ethers, combinations of two or more of the solvents, or mixtures of the above solvents or combinations with water. Wherein, the alcohols are selected from alcohols having 1-8 carbon atoms; the ketones are selected from ketones having 3-10 carbon atoms; the esters are selected from organic carboxylic acid esters; and the ethers are straight-chain or branched alkyl ethers or cyclic ether compounds.

6. The preparation method according to claim 5, characterized in that, The alcohols are methanol, ethanol, n-propanol, isopropanol, n-butanol, neopentyl alcohol, or a combination of two or more thereof; the ketones are acetone, butanone, pentanone, methyl ethyl ketone, 4-methyl-2-pentanone, or a combination of two or more thereof; the esters are methyl formate, ethyl acetate, isobutyl formate, ethyl propyl acetate, or a combination of two or more thereof; the ethers are methyl tert-butyl ether, tetrahydrofuran, 2-methyl-tetrahydrofuran, or a combination of two or more thereof.

7. The preparation method according to claim 4, characterized in that, The molar ratio of the compound of Formula I to the acid is 1:0.8 to 1:1.

5.

8. A single crystal of a monohydrochloride salt of the compound of formula I as defined in claim 3, characterized in that, The unit cell parameters of the single crystal are as follows: Orthorhombic crystal system, space group P 212121; a = 9.4704 (18) Å; b = 15.324 (4) Å; c = 17.437 (4) Å; V = 2530.5 (10) Å 3 ; Z = 4。 9. A crystal form A of a monohydrochloride salt of the compound of formula I as defined in claim 3, characterized in that, The crystal form A, when subjected to Cu-Kα radiation, exhibits characteristic peaks in X-ray powder diffraction at 9.66±0.20°, 16.08±0.20°, 18.10±0.20°, 21.30±0.20°, 21.68±0.20°, and 23.46±0.20° when expressed in 2θ angles.

10. The crystal form A according to claim 9, characterized in that, The crystal form A, when subjected to Cu-Kα radiation, exhibits characteristic peaks in X-ray powder diffraction at angles of 2θ at 9.66±0.20°, 11.62±0.20°, 16.08±0.20°, 18.10±0.20°, 21.30±0.20°, 21.68±0.20°, 23.46±0.20°, and 25.42±0.20°.

11. The crystal form A according to claim 9, characterized in that, The crystal form A is the anhydrous form of the monohydrochloride salt of compound I; Alternatively, the crystal form A has a DSC diagram as shown in Figure 2; Alternatively, the crystal form A has a TGA diagram as shown in Figure 3; Alternatively, crystal form A may be an irregularly shaped crystal; the grain size of crystal form A may not exceed 20 μm. Alternatively, the crystal form A has a PLM pattern as shown in Figure 4.

12. The crystal form A according to claim 9, characterized in that, The crystal form A, when subjected to Cu-Kα radiation, exhibits the following characteristic peaks in X-ray powder diffraction, expressed in 2θ angles, wherein the error range of the 2θ angles is ±0.20°: Alternatively, the crystal form A has a powder X-ray diffraction pattern as shown in Figure 1.

13. A crystal form B of a monohydrochloride salt of the compound of formula I as defined in claim 3, characterized in that, The crystal form B, when subjected to Cu-Kα radiation, exhibits characteristic peaks in X-ray powder diffraction at 9.48±0.20°, 15.44±0.20°, 18.10±0.20°, 19.80±0.20°, 22.10±0.20°, and 30.92±0.20° when expressed in 2θ angles.

14. The crystal form B according to claim 13, characterized in that, The crystal form B, when subjected to Cu-Kα radiation, exhibits characteristic peaks in X-ray powder diffraction at 9.48±0.20°, 10.78±0.20°, 15.44±0.20°, 18.10±0.20°, 19.18±0.20°, 19.80±0.20°, 22.10±0.20°, and 30.92±0.20°, as expressed in 2θ angles. Alternatively, the crystal form B is a monohydrate of the monohydrochloride salt of compound I; Alternatively, the crystal form B has a DSC diagram as shown in Figure 6; Alternatively, the crystal form B has a TGA diagram as shown in Figure 7; Alternatively, crystal form B is an irregularly shaped crystal; the grain size of crystal form B does not exceed 20 μm; Alternatively, the crystal form B has a PLM pattern as shown in Figure 8.

15. The crystal form B according to claim 13, characterized in that, The crystal form B, when subjected to Cu-Kα radiation, exhibits the following characteristic peaks in X-ray powder diffraction, expressed in 2θ angles, wherein the error range of the 2θ angles is ±0.20°: Alternatively, the crystal form B has a powder X-ray diffraction pattern as shown in Figure 5.

16. A crystal form C of a monohydrochloride salt of the compound of formula I as defined in claim 3, characterized in that, The crystalline form C, when subjected to Cu-Kα radiation, exhibits characteristic peaks in X-ray powder diffraction at 14.74±0.20°, 17.80±0.20°, 19.58±0.20°, 20.08±0.20°, 21.98±0.20°, 22.94±0.20°, and 25.92±0.20°, as expressed in 2θ angles.

17. The crystal form C according to claim 16, characterized in that, The crystalline form C, when subjected to Cu-Kα radiation, exhibits characteristic peaks in X-ray powder diffraction at 14.74±0.20°, 17.80±0.20°, 19.58±0.20°, 20.08±0.20°, 21.98±0.20°, 22.94±0.20°, 25.92±0.20°, and 33.48±0.20°, as expressed in 2θ angles.

18. The crystal form C according to claim 16, characterized in that, The crystal form C is the anhydrous form of the monohydrochloride salt of compound I; Alternatively, the crystal form C has a DSC diagram that is basically as shown in Figure 10; Alternatively, the crystal form C has a TGA diagram that is basically as shown in Figure 11; Alternatively, crystal form C is an irregularly shaped crystal; the grain size of crystal form C does not exceed 20 μm; Alternatively, the crystal form C has a PLM pattern as shown in Figure 12.

19. The crystal form C according to claim 16, characterized in that, The crystal form C, when subjected to Cu-Kα radiation, exhibits the following characteristic peaks in X-ray powder diffraction, expressed in 2θ angles, wherein the error range of the 2θ angles is ±0.20°: Alternatively, the crystal form C has a powder X-ray diffraction pattern as shown in Figure 9.

20. A crystal form D of a monohydrochloride salt of the compound of formula I as defined in claim 3, characterized in that, The crystal form D, when subjected to Cu-Kα radiation, exhibits characteristic peaks in X-ray powder diffraction at 10.16±0.20°, 11.90±0.20°, 15.74±0.20°, 16.58±0.20°, 19.22±0.20°, 20.24±0.20°, 21.98±0.20°, and 23.82±0.20°, as expressed in 2θ angles.

21. The crystal form D according to claim 20, characterized in that, The crystal form D is a monodichloromethane solvate of the monohydrochloride salt of compound I; Alternatively, the crystal form D has a DSC diagram as shown in Figure 14; Alternatively, the crystal form D has a TGA diagram as shown in Figure 15; Alternatively, crystal form D is an irregularly shaped crystal; the grain size of crystal form C does not exceed 10 μm; Alternatively, the crystal form D has a PLM pattern as shown in Figure 16.

22. The crystal form D according to claim 20, characterized in that, The crystal form D, when subjected to Cu-Kα radiation, exhibits the following characteristic peaks in X-ray powder diffraction, expressed in 2θ angles, wherein the error range of the 2θ angles is ±0.20°: Alternatively, the crystal form D has a powder X-ray diffraction pattern as shown in Figure 13.

23. A crystal form E of a monohydrochloride salt of the compound of formula I as defined in claim 3, characterized in that, The crystal form E, when subjected to Cu-Kα radiation, exhibits characteristic peaks in X-ray powder diffraction at 7.20±0.20°, 9.36±0.20°, 15.22±0.20°, 16.88±0.20°, 21.10±0.20°, 22.10±0.20°, and 23.68±0.20°, as expressed in 2θ angles.

24. The crystal form E according to claim 23, characterized in that, The crystal form E, when subjected to Cu-Kα radiation, exhibits characteristic peaks in X-ray powder diffraction at 7.20±0.20°, 9.36±0.20°, 15.22±0.20°, 16.88±0.20°, 18.78±0.20°, 21.10±0.20°, 22.10±0.20°, 23.68±0.20°, 26.04±0.20°, and 27.86±0.20°, as indicated by the 2θ angle. Alternatively, the crystal form E is an isopropanol solvate of the monohydrochloride salt of compound I.

25. The crystal form E according to claim 23, characterized in that, The crystal form E, when subjected to Cu-Kα radiation, exhibits the following characteristic peaks in X-ray powder diffraction, expressed in 2θ angles, wherein the error range of the 2θ angles is ±0.20°: Alternatively, the crystal form E has a powder X-ray diffraction pattern as shown in Figure 17.

26. A method for preparing crystal form A, crystal form B, crystal form C, crystal form D or crystal form E according to any one of claims 9-25, characterized in that: The method for preparing crystal form A is selected from one of the following methods: Method 1: Dissolve the compound of formula I in an alcohol solvent, add a solution of HCl in the alcohol solvent to form a salt, and then add n-alkane to crystallize, to obtain the crystal form A; The alcohol solvent is selected from ethanol and / or isopropanol; The n-alkane is selected from n-hexane and / or n-heptane; The mass-to-volume ratio of the compound of formula I, the alcohol solvent, and the n-alkane is 1 g : (10-30) mL : (10-30) mL; The concentration of the HCl solution in the alcohol solvent is 1-3 mol / L; Method 2: The monohydrochloride salt of compound I is heated and stirred in an alcohol solvent and n-alkane until dissolved, and then crystallized to obtain the crystal form A; The alcohol solvent is selected from ethanol and / or isopropanol; The n-alkane is selected from n-hexane and / or n-heptane; The mass-to-volume ratio of the monohydrochloride salt of the compound of formula I, the alcohol solvent, and the n-alkane is 1 g : (10-30) mL : (10-30) mL; The heating temperature is 45-75℃; The method for preparing crystal form B is as follows: crystal form A is placed under high humidity conditions to obtain crystal form B; The high humidity conditions are characterized by a temperature of 30-50℃ and a humidity of 60%-98%. The method for preparing the crystal form C is as follows: dissolve the compound of formula I in an alcohol solvent, then add a solution of HCl in the alcohol solvent to form a salt, and then add an ether solvent or an ester solvent to crystallize, thereby obtaining crystal form C; The alcohol solvent is selected from methanol, ethanol, or isopropanol; The ether solvent is selected from dimethyl ether, diethyl ether, propyl ether, or methyl tert-butyl ether; The ester solvent is selected from ethyl acetate or isopropyl acetate; The method for preparing crystal form D is as follows: the monohydrochloride salt of compound I is suspended and stirred in a haloalkane at room temperature to crystallize, thereby obtaining crystal form D; The haloalkane is selected from dichloromethane, trichloromethane, or carbon tetrachloride; The mass-to-volume ratio of the monohydrochloride salt of the compound of formula I to the haloalkane is 1 g:(15-35) mL; The method for preparing the crystal form E is as follows: the monohydrochloride salt of compound I is suspended and stirred in an alcohol solvent at room temperature to crystallize, thereby obtaining crystal form E; The alcohol solvent is selected from methanol, ethanol, or isopropanol; The mass-to-volume ratio of the monohydrochloride salt of the compound of formula I to the alcohol solvent is 1 g:(15-35) mL.

27. The preparation method according to claim 26, characterized in that, In the method for preparing crystal form C, The mass-to-volume ratio of the compound of formula I, the alcohol solvent, and the ether solvent is 1 g : (2-8) mL : (20-40) mL; The concentration of the HCl solution in the alcohol solvent is 1-3 mol / L; The mass ratio of the compound of formula I and the solution of HCl in an alcohol solvent is 1 g : (0.5-1.5) g.

28. A pharmaceutical composition comprising at least one of a pharmaceutically acceptable salt, a single crystal, a crystal form A, a crystal form B, a crystal form C, a crystal form D, and a crystal form E of a compound of formula I as claimed in any one of claims 1-3 or 8-25, and optionally a pharmaceutically acceptable excipient.

29. Use of at least one of the pharmaceutically acceptable salts, single crystals, crystal forms A, B, C, D, E, or pharmaceutical compositions of claim 28 of any one of the compounds of formula I according to any one of claims 1-3 or 8-25 in the preparation of a medicament for the prevention and / or treatment of complement factor B-mediated diseases or conditions.

30. The use as described in claim 29, wherein, The diseases or conditions mediated by complement factor B are selected from at least one of the following: paroxysmal nocturnal hemoglobinuria (PNH), primary glomerulonephritis (IgAN), membranous nephropathy (MN), C3 glomerulonephritis (C3G), age-related macular degeneration (AMD), geographic atrophy (GA), atypical hemolytic uremic syndrome (aHUS), hemolytic uremic syndrome (HUS), diabetic retinopathy (DR), hemodialysis complications, hemolytic anemia, neuromyelitis (NMO), arthritis, rheumatoid arthritis, liver inflammation, dermatomyositis and amyotrophic lateral sclerosis, myasthenia gravis (MG), respiratory diseases, and cardiovascular diseases.

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