BR112018015881B1 - TLR7 AGONIST MALEATE SALT, CRYSTALLINE FORMS, C, D AND E, THEIR PREPARATION PROCESSES AND USES, AND CRYSTALLINE AND PHARMACEUTICAL COMPOSITIONS - Google Patents

Abstract

The present invention relates to a maleate salt of a compound represented by formula I, a method for preparing the salt, a pharmaceutical composition containing the salt, and the use of the salt. The present invention likewise relates to crystalline forms C, D and E of the maleate salt of the compound represented by formula I, methods for preparing the crystalline forms, crystalline compositions and pharmaceutical compositions containing the crystalline forms, and use thereof.

Description

Technical field

[001] The present invention relates to the field of medicinal chemistry, and particularly relates to a TLR7 agonist maleate, a process for preparing the same, a pharmaceutical composition comprising the same and use thereof. The present invention likewise relates to crystalline form C, crystalline form D and crystalline form E of the salt, preparation processes, crystalline compositions comprising the crystalline forms, pharmaceutical compositions comprising the crystalline forms or crystalline compositions and use thereof.

BACKGROUND

[002] Similar Toll receptor is expressed by various immune cells and recognizes high reserved structural motifs: Pathogen-Associated Molecular Pattern (PAMP) expressed by microorganism pathogens or Damage-Associated Molecular Patterns (DAMP) released by dead cells. PAMP or DAMP stimulates Toll receptor-like signal cascade that induces activations of transcriptional factors such as AP-1, NF-KB and interferon regulators (pulse response function). It results in several cellular responses, including production of interferons, pro-inflammatory cytokines and effector cytokines, by which an immune response is produced. Undoubtedly, 13 types of similar Toll receptor have been found in mammals. Toll-like receptors 1, 2, 4, 5 and 6 are mainly expressed on the cell surface while Toll-like receptors 3, 7, 8 and 9 are expressed in the endosome. Different Toll-like receptors recognize ligands derived from different pathogens. Toll-like receptor 7 (TLR7) is mainly expressed by plasmacytoid dendritic cells (pDC), and recognized by ligand to induce the secretion of interferon α (IFN-α). Toll-like receptor 7 (TLR7) and Toll-like receptor 8 (TLR8) are highly homologous and therefore the ligand of TLR7 in most cases is likewise that of TLR8. TLR8 stimulation mainly induces the productions of cytokines such as tumor necrosis factor α (TNF-α) and chemoattractant. Interferon α is one of the drugs to treat chronic hepatitis B or hepatitis C while TNF-α is a pro-inflammatory cytokine, overexpression of which will result in serious side effects.

[003] Several TLR7 agonists have been reported, such as imiquimod (British Journal of Dermatology 2003; 149 (Suppl. 66): 5-8), resiquimod (Antiviral Research 64 (2004) 79-83), GS-9620 (Gastroenterology ( 2013), 144(7), 1508-1517). However, it is desirable to have new TLR7 agonists with better selectivity, activity and safety.

[004] Chinese patent application No. 201410405136.0 which is incorporated herein in its entirety by reference describes a small molecule, that is, 2-butoxy-7-(4-(pyrrolidin-1-ylmethyl)benzyl)-5H-pyrrolo[3 ,2-d]pyrimidin-4-amine and preparation process thereof having the following structure:

summary

[005] In one aspect, provided is a maleate of the compound of Formula I:

[006] In another aspect, provided are crystalline forms of the maleate of the compound of Formula I, the corresponding preparation process and the crystalline composition. In one embodiment, the crystal form is crystal form C, crystal form D, or crystal form E.

[007] In one embodiment, crystalline form C is characterized by an X-ray powder diffraction (XRPD) pattern having diffraction peaks at 2θ=7.6°±0.2°, 9.9°±0, 2°, 17.8°±0.2°, 22.8°±0.2°, 24.2°±0.2°, 26.3°±0.2°.

[008] In one embodiment, crystalline form D is characterized by an X-ray powder diffraction (XRPD) pattern having diffraction peaks at 2θ=5.1°±0.2°, 9.5°±0, 2°, 11.2°±0.2°, 17.6°±0.2°, 20.2°±0.2°, 23.0°±0.2°.

[009] In one embodiment, crystalline form E is characterized by an X-ray powder diffraction (XRPD) pattern having diffraction peaks at 2θ=4.9°±0.2°, 5.3°±0, 2°, 9.0°±0.2°, 16.5°±0.2°, 19.3°±0.2°.

[0010] In another aspect, provided is a pharmaceutical composition, comprising one or more crystalline forms or the crystalline composition thereof according to the invention. The pharmaceutical composition may also optionally comprise a pharmaceutically acceptable carrier, excipient and/or medium.

[0011] In another aspect, provided is a method for treating or preventing disease associated with Toll-like receptor 7 (TLR7), comprising administering to an individual in need thereof the crystalline form or the crystalline composition or the pharmaceutical composition according to the invention in an effective amount. Preferably, the disease is virus infection.

[0012] In another aspect, provided is use of the crystalline form or the crystalline composition or the pharmaceutical composition according to the invention for the manufacture of a medicament for treating or preventing disease associated with Toll-like receptor 7 (TLR7). Preferably, the disease is virus infection.

[0013] In yet another aspect, provided is the crystalline form or crystalline composition or pharmaceutical composition according to the invention for use in the treatment or prevention of disease associated with Toll-like receptor 7 (TLR7). Preferably, the disease is virus infection.

[0014] In one embodiment of the invention, the virus infection is hepatitis virus infection, particularly hepatitis B or hepatitis C virus infection.

Brief description of the drawings

[0015] Figure 1: XRPD pattern of the crystalline form C of the maleate of the compound of Formula I.

[0016] Figure 2: XRPD pattern of the crystalline form D of the maleate of the compound of Formula I.

[0017] Figure 3: XRPD pattern of the crystalline form E of the maleate of the compound of Formula I.

Detailed Description General definition and terminology

[0018] Unless otherwise stated, the terms and phrases used herein have the following meanings. A specific term or phrase will not be considered as unclear or undefined when it is not specifically defined. It should be understood according to the general meaning. The trade name used here refers to the corresponding product or active ingredient.

[0019] Unless otherwise specifically defined, proportion (including percentage) or part is calculated based on weight here.

[0020] When used with a numerical variable, the term “approximate” or “about” typically refers to the value of the variable and all values of the variable within experimental error (e.g., within a mean confidence interval of 95 %) or within ±10% of the specified value, or a wider range.

[0021] The expression “comprises” or its synonyms “contain”, “include”, “have” or similar is open-ended that does not exclude other elements, steps or ingredients not listed. The expression “consists of” excludes any element, steps or ingredients not listed. The expression “substantially consists of” refers to specified elements, steps or ingredients within a certain range, together with optional elements, steps or components that do not substantially affect the basic and novel characteristic of the claimed subject matter. It should be understood that the expression “comprises” encompasses the expressions “substantially consists of” and “consists of”.

[0022] The term “optional” or “optionally” means that the event described below may or may not happen. This term covers cases in which the event may or may not happen.

[0023] The term “pharmaceutical composition” refers to an active ingredient that is optionally combined with one or more pharmaceutically acceptable components (for example, but not limited to carrier and/or excipient). The active ingredient is exemplified as the compound of Formula I or maleate thereof, one or more crystalline forms according to the invention, or one or more crystalline compositions according to the invention.

[0024] The term “pharmaceutically acceptable vehicle” refers to those vehicles that do not have significant irritation and do not impair the bioactivity and properties of the active compound. “Pharmaceutically acceptable carrier” refers to an inert substance that is administered with an active ingredient and is beneficial to the administration thereof, and includes, but is not limited to, any of the following substances approved by the State Food and Drug Administration for use in humans or animals (e.g. cattle example): glidant, sweetening agent, diluent, preservative, dye/colorant, flavoring agent, surfactant, wetting agent, dispersant, disintegrant, suspending agent, stabilizing agent, isotonic agent, solvent or emulsifying agent. Non-limiting examples of the carriers include calcium carbonate, calcium phosphate, various sugars and starches, cellulose derivatives, gelatin, vegetable oil and polyethylene glycol or the like. Other information regarding vehicles can be found in Remington: The Science and Practice of Pharmacy, 21st Ed., Lippincott, Williams & Wilkins (2005) of which the contents are incorporated herein by reference. The term “excipient” generally refers to the vehicle, diluent and/or medium used to formulate effective pharmaceutical composition.

[0025] The term “administration” or “administering” or similar refers to a method that allows a compound or composition to be delivered to a desired site of biological action. Such methods include, but are not limited to, oral, parenteral (including intravenous, subcutaneous, intraperitoneal, intramuscular, intravascular injection or infusion), local, rectal or similar administration.

[0026] As for an active pharmaceutical or pharmacological agent, the term “effective amount” refers to the amount of the medicine or agent that is not toxic, but sufficient to achieve the desired effect. With respect to the oral formulation herein, the “effective amount” for an active substance in the composition refers to the amount required to obtain the desired effect in combination with another active substance in the composition. The effective amount can be determined individually and depends on the age and general condition of the recipient as well as the specific active substance. The effective amount in a specific case can be determined by a person skilled in the art by conventional testing.

[0027] The term “active ingredient”, “therapeutic agent”, “active substance” or “active agent” refers to a chemical entity useful for treating or preventing target disorder, disease or condition effectively. The term may here refer to for example the compound of Formula I or maleate thereof, one or more crystalline forms according to the invention or one or more crystalline compositions according to the invention.

[0028] In X-ray powder diffraction (XRPD or XRD) spectra, the diffraction pattern obtained from a crystalline compound is generally characteristic for a particular crystalline form in which the relative intensities of the bands (especially at low angles) may vary with the dominant orientation effect due to the difference in crystallization conditions, particle diameters, and other measurement conditions. Therefore, the relative intensities of diffraction peaks are not characteristic for the given crystal form. It is more important to note the relative positions of peaks rather than their relative intensities when determining whether the crystalline form is the same as that known in the art. Furthermore, there may be slight errors in peak positions for any given crystalline form that is also well known in the art of crystallography. For example, peak position may shift due to change in temperature, sample movement, or instrument calibration during sample analysis; and the measurement error of 2θ value can sometimes be about ±0.2°, typically about ±0.1°. Therefore, this error should be taken into account when determining the crystal structure. If the crystalline forms according to the invention are described as substantially as shown in the figure, the term “substantially” is likewise intended to encompass such differences in the diffraction peak.

[0029] In the XRPD pattern, the peak position is normally represented by angle 2θ or crystal surface distance d and a simple conversion between d and θ is d =À /2sinθ, where d represents the crystal surface distance, À represents the length of incident X-ray wave, and θ is diffraction angle. As for the same crystalline form of the same compound, the peak positions of the XRPD pattern are similar as a whole, and the relative intensity error can be large. It should also be noted that, in identifying a mixture, some diffraction lines may be lost due to factors such as decrease in content, etc., and therefore it is not necessary to rely on the entire bands observed in the high purity sample, and yet a band may be characteristic for a particular crystal.

[0030] Differential scanning calorimetry (DSC) is used to measure the transition temperature when a crystal absorbs or releases heat due to changes in its crystal structure or crystal melting. The thermal transition temperature and melting point error is typically within about 5°C, typically about 3°C for the same crystalline form of the same compound in a continuous analysis. When a compound is described as having a particular DSC peak or melting point, it means the DSC peak or melting point ±5°C. Provided is an auxiliary DSC method to identify different crystalline forms. Different crystalline forms can be identified according to their different transition temperature characteristics. It is noted that the DSC peak or melting point of the mixture can vary over a wide range. Furthermore, melting temperature is associated with the rate of temperature rise due to decomposition during melting of the substance.

[0031] Differential scanning calorimetry (DSC) here is measured by the following method: device: TA Q2000 differential scanning calorimeter; method: a sample (~1 mg) is placed in a DSC aluminum pan, method: 25°C~300°C, heating rate 10°C/min.

[0032] The term “crystalline composition” refers to a solid form, which comprises one or more crystalline forms according to the invention (such as crystalline form C, D and/or E). The amounts of crystalline forms contained in the crystalline composition may be 50% or more, 80% or more, 90% or more, or 95% or more independently. In addition to the crystalline form according to the invention, the crystalline composition may also optionally comprise another crystal or amorphous form of the compound of Formula I or the salt thereof (such as maleate) or impurities other than these substances. It should be understood by those skilled in the art that the sum of the contents of the components in the crystalline composition should be 100%.

Formula I compound maleate

[0033] Provided is a maleate of the compound of Formula I:

[0034] In a preferred embodiment, in maleate of the compound of Formula I, the molar ratio of the compound of Formula I to maleic acid is 1:1~3, preferably 1:2. The maleate of the compound of Formula I can be prepared by conventional maleate preparation process.

Crystalline form C

[0035] Provided is maleate crystalline form C of the compound of Formula I which have diffraction peaks at 2θ=7.6°±0.2° 9.9°±0.2°, 17.8°±0.2 °, 22.8°±0.2°, 24.2°±0.2°, 26.3°±0.2° in X-ray diffraction pattern (XRPD).

[0036] In a specific embodiment, crystalline form C has diffraction peaks at 2θ=5.6°±0.2°, 7.6°±0.2°, 9.9°±0.2°, 17 .8°±0.2°, 19.8°±0.2°, 22.8°±0.2°, 24.2°±0.2°, 25.0°±0.2°, 26 .3°±0.2° in X-ray diffraction pattern (XRPD).

[0037] In a more specific embodiment, crystalline form C has diffraction peaks at 2θ=5.6°±0.2°, 6.0°±0.2°, 7.6°±0.2°, 9.9°±0.2°, 12.0°±0.2°, 15.3°±0.2°, 17.8°±0.2°, 18.5°±0.2°, 19.8°±0.2°, 20.4°±0.2°, 22.8°±0.2°, 23.1°±0.2°, 24.2°±0.2°, 24.7°±0.2°, 25.0°±0.2°, 26.3°±0.2° in X-ray diffraction pattern (XRPD).

[0038] In a particular embodiment, diffraction peaks of the crystalline form C of the maleate of the compound of Formula I are characterized as follows:

[0039] In one embodiment, the X-ray powder diffraction pattern of the crystalline form C of the maleate of the compound of Formula I is substantially shown in Figure 1.

[0040] Crystalline form C can also be characterized by DSC, with an initial temperature of 97.0°C±5°C and a peak temperature of 10.0°C±5°C.

[0041] In one embodiment, in crystalline form C, the molar ratio of the compound of Formula I to maleic acid is 1:1~3, preferably 1:2.

[0042] Also provided is a process for preparing crystalline form C according to the invention, comprising precipitating the maleate of the compound of Formula I from a solvent.

[0043] In one embodiment, the process comprises the following steps: 1) dissolving the compound of Formula I in a solvent that is preferably heated to promote dissolution; 2) add maleic acid; and 3) cool for crystallization, filtering, washing and drying to obtain crystalline form C.

[0044] In step 1), the solvent is selected from the group consisting of methanol, ethanol, propanol, isopropanol, n-butanol, isobutanol, tertiary butanol, acetone, ethyl acetate and mixed solvent thereof; preferably ethanol.

[0045] In step 1), the amount of solvent added per 1 mole of the compound of Formula I is 0.2 to 8L, preferably 0.3 to 4L, more preferably 0.5 to 2L, preferably 1L.

[0046] In step 1), the heating temperature can be 40°C to 90°C, preferably 50°C to 80°C, more preferably 70°C to 80°C.

[0047] In step 2), the amount of maleic acid added per 1 mol of the compound of Formula I is 1.0 to 4.0 mol, preferably 1.6 to 3.0 mol, more preferably 2.0 to 2. 4 mol, preferably 2.2 mol.

[0048] Also provided is a crystalline composition comprising crystalline form C. In one embodiment, based on the weight of the crystalline composition, the content of crystalline form C is 50% or more, preferably 80% or more, more preferably 90 % or more, and preferably 95% or more. The crystalline composition, in addition to crystalline form C, may also comprise the compound of Formula I or its salt in other crystalline or amorphous forms, or impurities other than these substances.

Crystalline form D

[0049] Provided is crystalline form D of the maleate of the compound of Formula I which have diffraction peaks at 2θ=5.1°±0.2° 9.5°±0.2°, 11.2°±0.2 °, 17.6°±0.2°, 20.2°±0.2°, 23.0°±0.2° in X-ray diffraction pattern (XRPD).

[0050] In a specific embodiment, crystalline form D has diffraction peaks at 2θ=5.1°±0.2°, 9.5°±0.2°, 11.2°±0.2°, 17 .6°±0.2°, 20.2°±0.2°, 20.7°±0.2°, 23.0°±0.2°, 23.7°±0.2° in standard X-ray diffraction (XRPD).

[0051] In a more specific embodiment, crystalline form D has a diffraction peak at 2θ=5.1°±0.2°, 5.6°±0.2°, 9.5°±0.2°, 11.2°±0.2°, 16.9°±0.2°, 17.6°±0.2°, 20.2°±0.2°, 20.7°±0.2°, 22.6°±0.2°, 23.0°±0.2°, 23.7°±0.2°, 24.5°±0.2° in X-ray diffraction pattern (XRPD).

[0052] In a particular embodiment, the diffraction peaks of the crystalline form D of the maleate of the compound of Formula I are characterized as follows:

[0053] In one embodiment, the X-ray powder diffraction pattern of the crystalline form D of the maleate of the compound of Formula I is substantially shown in Figure 2.

[0054] The crystalline form that D can also be characterized by DSC, with an initial temperature of 98.3 °C±5 °C and a peak temperature of 110.1 °C±5 °C.

[0055] In one embodiment, in crystalline form D the molar ratio of the compound of Formula I to maleic acid is 1:1~3, preferably 1:2.

[0056] Also provided is a process for preparing crystalline form D, comprising the following steps: 1) placing the crystalline form C of the maleate of the compound of Formula I in acetone solvent to form a suspension; 2) stir at constant temperature; 3) centrifuge, wash and dry to obtain crystalline form D.

[0057] In step 1), the amount of acetone added per 1 g of crystalline form C of the maleate of the compound of Formula I is 2 to 30 mL, preferably 8 to 24 mL, more preferably 12 to 20 mL, preferably 14 to 16 mL.

[0058] In step 2), the constant temperature is 20°C to 60°C, preferably 30°C to 50°C, more preferably 35°C to 45°C, preferably 40°C.

[0059] Also provided is a crystalline composition comprising crystalline form D. In one embodiment, based on the weight of the crystalline composition, crystalline form D is 50% or more, preferably 80% or more, more preferably 90% or more, and preferably 95% or more. Furthermore, the crystalline composition, in addition to crystalline form D, may also comprise a compound of Formula I or salt thereof in other crystalline or amorphous forms, or impurities other than these substances.

Crystalline form E

[0060] Provided is crystalline form E of the maleate of the compound of Formula I which have diffraction peaks at 2θ=4.9°±0.2° 5.3°±0.2°, 9.0°±0.2 °, 16.5°±0.2°, 19.3°±0.2° in X-ray diffraction pattern (XRPD).

[0061] In a specific embodiment, crystalline form E has diffraction peaks at 2θ=4.9°±0.2°, 5.3°±0.2°, 6.7°±0.2°, 9 ,0°±0.2°, 10.8°±0.2°, 16.5°±0.2°, 19.3°±0.2° in X-ray diffraction pattern (XRPD).

[0062] In a more specific embodiment, crystalline form E has diffraction peaks at 2θ=4.9°±0.2°, 5.3°±0.2°, 6.7°±0.2°, 9.0°±0.2°, 10.8°±0.2°, 16.2°±0.2°, 16.5°±0.2°, 19.3°±0.2°, 22.0°±0.2°, 22.6°±0.2°, 25.9°±0.2° in X-ray diffraction pattern (XRPD).

[0063] In a specific embodiment, the diffraction peaks of the crystalline form E of the maleate of the compound of Formula I are characterized as follows:

[0064] In one embodiment, the X-ray powder diffraction pattern of the crystalline form E of the maleate of the compound of Formula I is substantially shown in Figure 3.

[0065] Crystalline form E can also be characterized by DSC with an initial temperature of 85.7°C±5°C and a peak temperature of 97.5°C±5°C.

[0066] In one embodiment, in crystalline form E, the molar ratio of the compound of Formula I to maleic acid is 1:1~3, preferably 1:2.

[0067] Also provided is a process for preparing crystalline form E, comprising the following steps: 1) placing the crystalline form C of the maleate of the compound of Formula I in isopropanol solvent to form a suspension; 2) stir at constant temperature; 3) centrifuge, wash and dry to obtain crystalline form E.

[0068] In step 1), the amount of isopropanol added per 1 g of the crystalline form C of the maleate of the compound of Formula I is 2 to 30 mL, preferably 8 to 24 mL, more preferably 12 to 20 mL, also preferably 14 to 16 mL.

[0069] In step 2), the constant temperature is 20°C to 60°C, preferably 30°C to 50°C, more preferably 35°C to 45°C, also preferably 40°C.

[0070] Also provided is a crystalline composition comprising crystalline form E. In one embodiment, based on the weight of the crystalline composition, crystalline form E is 50% or more, preferably 80% or more, more preferably 90% or more , and preferably 95% or more. The crystalline composition, in addition to crystalline form E, may also comprise the compound of Formula I or its salt in other crystalline or amorphous forms, or impurities other than these substances.

Pharmaceutical composition and administration

[0071] Provided is a pharmaceutical composition comprising maleate of the compound of Formula I; the crystalline form C or the crystalline composition comprising the crystalline form C; the crystalline form D or the crystalline composition comprising the crystalline form D; or the crystalline form E or the crystalline composition comprising the crystalline form E, or any combination thereof in an effective amount. Furthermore, the pharmaceutical composition may or may not comprise a pharmaceutically acceptable carrier, excipient and/or medium.

[0072] The compound according to the invention is administered in a pure form or in a suitable pharmaceutical composition form, which can be carried out by any acceptable mode of administration of the agent with similar use. Pharmaceutical composition according to the invention can be prepared by combining the compound according to the invention or the salt thereof with a suitable pharmaceutically acceptable carrier, for example it can be formulated in solid, semi-solid, liquid or gas formulation, such as tablet, pill, capsule, powder, granule, ointment, emulsion, suspension, solution, suppository, injection, inhalant, gel, microsphere, aerosol or similar.

[0073] The pharmaceutical composition according to the invention can be prepared by processes well known in the art, such as conventional mixing, dissolution, granulation, tablet coating, levigation, emulsion, freeze drying or the like.

[0074] Typical routines for administering the compound according to the invention or the pharmaceutical composition thereof comprise, but are not limited to, oral, rectal, transmucosal, enteral or local administration, transcutaneous, inhalant, parenteral, sublingual, intravaginal, intranasal, intraocular, intraperitoneal, intramuscular, subcutaneous, intravenous.

[0075] In a preferred embodiment, the pharmaceutical composition is in an oral administration form. As for oral administration, the active compounds can be mixed with pharmaceutically acceptable carriers, excipients and/or media well known in the art to prepare the pharmaceutical composition. The vehicles, excipients and media can be used to prepare the compounds according to the invention into a tablet, pill, troche, dragee, capsule, liquid, gel, paste, suspension or the like useful for oral administration to the patient.

[0076] Solid oral composition can be prepared by conventional mixing, loading or compression processes, for example, by the following processes: mixing the active compound with solid excipients, optionally grinding the resulting mixture, adding other own adjuvants if necessary, and then processing the mixture into granules to obtain the tablet core or dragee. The excipients themselves include, but are not limited to, fillers such as sugar, including lactose, sucrose, mannitol or sorbitol; preparing cellulose such as microcrystalline cellulose, corn starch, wheat starch, rice starch and potato starch; and other substances, such as silica gel, gelatin, tragacanth, methylcellulose, hydroxypropylmethyl cellulose, sodium carboxymethyl cellulose and/or polyvinylpyrrolidone; disintegrant, such as sodium carboxymethyl starch, croscarmellose sodium, cross-linked polyvinylpyrrolidone, agar or alginic acid. Salt such as sodium alginate can also be used. The core of the tablets may optionally be coated by processes well known in general pharmaceutical practice, especially by enteric coating.

Beneficial effects

[0077] The crystalline form C, crystalline form D and crystalline form E of the maleate of the compound of Formula I according to the invention have the advantages of high purity, high crystallinity and good stability which are suitable for manufacturing a medicine to prevent or treat Toll-like receptor 7 (TLR7)-associated disease.

[0078] The technical solutions of the invention are illustrated according to the following paragraphs [1] to [44]: [1] A maleate of the compound of Formula I, [2] The maleate of the compound of Formula I according to paragraph [1], characterized by the fact that the molar ratio of the compound of Formula I to maleic acid is 1:1~3, preferably 1:2. [3] A pharmaceutical composition, comprising the maleate of the compound of Formula I according to paragraph [1] or [2] in an effective amount. [4] Use of the maleate of the compound of Formula I according to paragraph [1] or [2] or the pharmaceutical composition according to paragraph [3] for the manufacture of a medicament for treating disease associated with Toll-like receptor 7. [5] The use according to paragraph [4], characterized by the fact that, the disease is virus infection, particularly hepatitis virus infection, for example hepatitis B or hepatitis C virus infection. [6] A form crystalline C of the maleate of the compound of Formula I, characterized by the fact that it has diffraction peaks at 2θ=7.6°±0.2°, 9.9°±0.2°, 17.8°±0.2° , 22.8°±0.2°, 24.2°±0.2°, 26.3°±0.2° in X-ray powder diffraction pattern. [7] The crystalline form C according to paragraph [6], characterized by the fact that it has diffraction peaks at 2θ=5.6°±0.2°, 7.6°±0.2°, 9.9°±0.2°, 17.8° ±0.2°, 19.8°±0.2°, 22.8°±0.2°, 24.2°±0.2°, 25.0°±0.2°, 26.3° ±0.2° in X-ray powder diffraction pattern. [8] The crystalline form C according to paragraph [7], characterized by the fact that it has diffraction peaks at 2θ=5.6°±0.2° , 6.0°±0.2°, 7.6°±0.2°, 9.9°±0.2°, 12.0°±0.2°, 15.3°±0.2° , 17.8°±0.2°, 18.5°±0.2°, 19.8°±0.2°, 20.4°±0.2°, 22.8°±0.2° , 23.1°±0.2°, 24.2°±0.2°, 24.7°±0.2°, 25.0°±0.2°, 26.3°±0.2° in X-ray powder diffraction pattern. [9] The crystalline form C according to any one of paragraphs [6][8], characterized by the fact that, has X-ray powder diffraction pattern substantially shown in Figure 1. [10] The crystalline form C according to any one of paragraphs [6]-[9], characterized by the fact that, when characterized by DSC, the initial temperature is 97.0°C±5°C and the peak temperature is 106.0°C±5°C. [11] A process for preparing crystalline form C according to any one of paragraphs [6]-[10], comprising the following steps: 1) dissolving the compound of Formula I in a solvent that is preferably heated to promote dissolution ; 2) add maleic acid; 3) cool for crystallization, filter, wash and dry to obtain crystalline form C. [12] The preparation process according to paragraph [11], characterized by the fact that the solvent in step 1) is selected from the group consisting of methanol, ethanol, propanol, isopropanol, n-butanol, isobutanol, tertiary butanol, acetone, ethyl acetate and mixed solvent thereof. [13] The preparation process according to paragraph [12], characterized by the fact that the solvent is ethanol. [14] The preparation process according to any one of paragraphs [11]-[13], characterized by the fact that, in step 1), the amount of solvent added per 1 mol of the compound of Formula I is 0.2 to 8L, preferably 0.3 to 4L, more preferably 0.5 to 2L, preferably 1L. [15] The preparation process according to any one of paragraphs [11]-[14], characterized by the fact that, in step 2), the amount of maleic acid added per 1 mol of the compound of Formula I is 1, 0 to 4.0 mol, preferably 1.6 to 3.0 mol, more preferably 2.0 to 2.4 mol, preferably 2.2 mol. [16] The preparation process according to any one of paragraphs [11]-[14], characterized by the fact that, in step 1), the heating temperature may be 40°C to 90°C, preferably 50° C to 80°C, more preferably 70°C to 80°C. [17] A crystalline composition, characterized in that, based on the weight of the crystalline composition, the crystalline form C according to any one of paragraphs [6]-[10] is 50% or more, preferably 80% or more , more preferably 90% or more, and preferably 95% or more. [18] A pharmaceutical composition, comprising the crystalline form C according to any one of paragraphs [6]-[10] or the crystalline composition according to paragraph [17] in an effective amount. [19] Use of the crystalline form C according to any of paragraphs [6]-[10] or the crystalline composition according to paragraph [17] or the pharmaceutical composition according to paragraph [18] for the manufacture of a medicament to treat Toll-like receptor 7 (TLR7)-associated disease. [20] The use according to paragraph [19], characterized by the fact that the disease is virus infection, particularly hepatitis virus infection, such as hepatitis B virus infection or hepatitis C virus infection. [21] A crystalline form D of the maleate of the compound of Formula I, characterized by the fact that it has diffraction peaks at 2θ=5.1°±0.2°, 9.5°±0.2°, 11.2°±0.2°, 17.6°±0.2°, 20.2°±0.2°, 23.0°±0.2° in X-ray powder diffraction pattern. [22] The crystalline form D according to paragraph [21] characterized by the fact that it has diffraction peaks at 2θ=5.1°±0.2°, 9.5°±0.2°, 11.2°±0.2°, 17.6°±0 .2°, 20.2°±0.2°, 20.7°±0.2°, 23.0°±0.2°, 23.7°±0.2° in powder diffraction pattern X-ray. [23] The crystalline form D according to paragraph [22], characterized by the fact that it has diffraction peaks at 2θ=5.1°±0.2°, 5.6°±0.2°, 9 .5°±0.2°, 11.2°±0.2°, 16.9°±0.2°, 17.6°±0.2°, 20.2°±0.2°, 20 .7°±0.2°, 22.6°±0.2°, 23.0°±0.2°, 23.7°±0.2°, 24.5°±0.2° in standard X-ray powder diffraction pattern. [24] The crystalline form D according to any one of paragraphs [21]-[23], characterized by the fact that the crystalline form D has an X-ray powder diffraction pattern substantially shown in Figure 2. [25] The crystalline form D according to any one of paragraphs [21]-[24], characterized by the fact that, when characterized by DSC, the initial temperature is 983°C ±5°C and the peak temperature is 1101°C ±5°C. [26] A process for preparing crystalline form D according to paragraph [25], comprising the following steps: 1) placing crystalline form C of the maleate of the compound of Formula I in acetone solvent to form a suspension; 2) stir at constant temperature; 3) centrifuge, wash and dry to obtain crystalline form D. [27] The preparation process according to any of paragraphs [21]-[26], characterized by the fact that, in step 1), the amount of Acetone added per 1 g of maleate crystalline form C of the compound of Formula I is 2 to 30 ml, preferably 8 to 24 ml, more preferably 12 to 20 ml, preferably 14 to 16 ml. [28] The preparation process according to any one of paragraphs [21]-[27], characterized by the fact that, in step 2), the constant temperature is 20°C to 60°C, preferably 30°C for 50°C, more preferably 35°C to 45°C, preferably 40°C. [29] A crystalline composition, characterized by the fact that, based on the weight of the crystalline composition, the crystalline form D according to any one of paragraphs [21]-[25] is 50% or more, preferably 80% or more , more preferably 90% or more, and preferably 95% or more. [30] A pharmaceutical composition, comprising the crystalline form D according to any one of paragraphs [21]-[25] or the crystalline composition according to paragraph [29] in an effective amount. [31] Use of the crystalline form D according to any of paragraphs [21]-[25] or the crystalline composition according to paragraph [29] or the pharmaceutical composition according to paragraph [30] for the manufacture of a medicament to treat Toll-like receptor 7 (TLR7)-associated disease. [32] The use according to paragraph [31], characterized by the fact that the disease is virus infection, particularly hepatitis virus infection, such as hepatitis B or hepatitis C virus infection. [33] A crystalline form And the compound maleate of Formula characterized by the fact that it has diffraction peaks at 2θ=4.9°±0.2°, 5.3°±0.2°, 9.0°±0.2°, 16, 5°±0.2°, 19.3°±0.2° in X-ray powder diffraction pattern. [34] The crystalline form E according to paragraph [33] characterized by the fact that it has diffraction peaks at 2θ=4.9°±0.2°, 5.3°±0.2°, 6.7°±0.2°, 9.0°±0.2°, 10.8°±0.2 °, 16.5°±0.2°, 19.3°±0.2° in X-ray powder diffraction pattern. [35] The crystalline form E according to paragraph [34], characterized by the fact that have diffraction peaks at 2θ=4.9°±0.2°, 5.3°±0.2°, 6.7°±0.2°, 9.0°±0.2°, 10.8 °±0.2°, 16.2°±0.2°, 16.5°±0.2°, 19.3°±0.2°, 22.0°±0.2°, 22.6 °±0.2°, 25.9°±0.2° in X-ray powder diffraction pattern. [36] The crystalline form E according to any of paragraphs [33]-[35], characterized by the fact that, the crystalline form E has X-ray powder diffraction pattern substantially shown in Figure 3. [37] The crystalline form E according to any of paragraphs [33]-[36], characterized by the fact that , when characterized by DSC, the initial temperature is 85.7 °C ±5 °C and the peak temperature is 97.5 °C ±5 °C. [38] A process for preparing crystalline form E according to any one of paragraphs [33]-[37], comprising the following steps: 1) placing crystalline form C of the maleate of the compound of Formula I in isopropanol solvent to form a suspension; 2) stir at constant temperature; 3) centrifuge, wash and dry to obtain crystalline form E. [39] The preparation process according to paragraph [38], characterized by the fact that, in step 1), the amount of isopropanol added per 1 g of crystalline form C of the maleate of the compound of Formula I is 2 to 30 ml, preferably 8 to 24 ml, more preferably 12 to 20 ml, also preferably 14 to 16 ml. [40] The preparation process according to paragraphs [38] or [39], characterized by the fact that, in step 2), the constant temperature is 20°C to 60°C, preferably 30°C to 50° C, more preferably 35°C to 45°C, also preferably 40°C. [41] A crystalline composition, characterized by the fact that, based on the weight of the crystalline composition, the crystalline form E according to any one of paragraphs [33]-[37] is 50% or more, preferably 80% or more , more preferably 90% or more, and preferably 95% or more. [42] A pharmaceutical composition, comprising the crystalline form E according to any one of paragraphs [33]-[37] or the crystalline composition according to paragraph [41] in an effective amount. [43] Use of the crystalline form E according to any of paragraphs [33]-[37] or the crystalline composition according to paragraph [41] or the pharmaceutical composition according to paragraph [42] for the manufacture of a medicament to treat Toll-like receptor 7 (TLR7)-associated disease. [44] The use according to paragraph [43], characterized by the fact that the disease is virus infection, particularly hepatitis virus infection, such as hepatitis B or hepatitis C virus infection.

Examples

[0079] The following abbreviations are used here: SEM-Cl: 2-(trimethylsilyl)ethoxymethyl chloride; SEM: 2-(trimethylsilyl)ethoxymethyl; DIPEA: diisopropylethylamine; TFA: trifluoroacetic acid; DMF: N,N-dimethylformamide; n-BuOH: n-butanol; NH3^O: aqueous ammonia; Na: sodium; XRPD: X-ray powder diffraction; DSC: differential thermal analysis.

[0080] In the present invention, the X-ray diffraction pattern was determined as follows: apparatus: Bruker D8 ADVANCED X-ray diffractometer; method: target: Cu: K-alpha; wavelength À =1.54179Â; voltage: 40 kV; current: 40 MA; scanning range: 4~40°; sample rotation speed: 15 rpm; scanning speed: 10°/min.

[0081] The solvents used here are commercially available and can be used without further purification. Synthesis reactions in preparation examples are generally carried out under an inert nitrogen atmosphere in anhydrous solvent. Proton MRI data is recorded on a Bruker Avance III 400 (400 MHz) spectrometer, with the chemical exchange shown as (ppm) in low-field tetramethylsilane. Mass spectrometry is determined on Agilent 1200 plus 6110 (&1956A). LC/MS or Shimadzu MS includes a DAD: SPD-M20A(LC) and Shimadzu Micromass 2020 detector. Mass spectrometer is equipped with an electrospray ionization (ESI) operated in positive or negative mode. Preparation Example 1: Preparation of 2-butoxy-7-(4-(pyrrolidin-1-ylmethyl)benzyl)-5H-pyrrolo[3,2-d]pyrimidin-4-amine maleate Formula III: 2,4- dichloro-5-((2-(trimethylsilyl)ethoxy)methyl)-5H-pyrrolo[3,2-d]pyrimidine

[0082] The compound of Formula II (2,4-dichloro-5H-pyrrolo[3,2-d]pyrimidine) (4.00 kg, 21.28 mol) was dissolved in DMF (20.00 L), and DIPEA (2.58 kg, 20.00 mol) was added in portions at room temperature (25°C) followed by stirring for 30 minutes. The reaction liquid was cooled to 0°C with an ice bath, and then SEM-Cl (4.00 kg, 24.00 mol) was added dropwise slowly over 5 hours at a drip rate of 1 to 2 drops. per second. After addition, the reaction liquid was stirred at 0°C for 4 hours. The reaction was monitored by HPLC. After completion, the reaction liquid was quenched and diluted with 70 L of water and then extracted with ethyl acetate (15 L*3). The combined organic phase was washed successively with 1 M aqueous hydrochloric acid (5 L*2) and saturated brine (7 L*2), and the solvent was distilled off under reduced pressure to yield the compound of Formula III (6. 40 kg, 20.11 mol, yield 94.50%). 1H NMR (400 MHz, DMSO-d6) δ 8.24 - 8.35 (m, 1 H), 6.70 - 6.85 (m, 1 H), 5.77 (s, 2 H), 3 .45 - 3.57 (m, 2 H), 0.74 - 0.86 (m, 2 H), 0.00 (s, 9 H). Formula IV: 2-chloro-5-((2-(trimethylsilyl)ethoxy)methyl)-5H-pyrrolo[3,2-d] pyrimidin-4-amine

[0083] The compound of Formula III (1.60 kg, 5.03 mol) was dissolved in isopropanol (1.60 L) in a 10 L flask. Aqueous ammonia (4 L) was added at room temperature (25° C) in one portion and the reaction mixture was stirred at 95°C for 7 hours. The reaction was monitored by HPLC. Upon completion, the reaction liquid was allowed to cool to room temperature and filtered through a Buchner funnel to yield a dark brown solid. The solid was successively suspended with ethyl acetate/n-heptane (1/1.5 L*2) and ethyl acetate (4 L) to yield the compound of Formula IV as a brown solid (1.25 kg, 4. 18 mol, yield 83.1%). 1H NMR (400 MHz, DMSO-d6) δ 7.61 - 7.77 (m, 1 H), 6.97 - 7.19 (m, 2 H), 6.28 - 6.38 (m, 1 H), 5.54 - 5.67 (m, 2 H), 3.43 - 3.53 (m, 2 H), 0.76 - 0.91 (m, 2 H), 0.07 (s , 9 H). Formula V: 2-butoxy-5-((2-(trimethylsilyl)ethoxy)methyl)-5H-pyrrolo[3,2-d]pyrimidin-4-amine

[0084] To n-BuOH (17.0 L) metal sodium (525.05 g, 22.84 mol) was slowly added in portions under nitrogen. After addition, the temperature of the system was raised to 60 °C, and stirring was carried out continuously at the temperature until the sodium metal was completely dissolved. Then the system was cooled to 25°C, and the compound of Formula IV (1.95 kg, 6.53 mol) was added in portions. After being homogeneously mixed with stirring, the reaction mixture was continuously stirred for 8 h at 90°C. The reaction was monitored by HPLC. After completion, the reaction mixture was allowed to cool spontaneously to 25 ° C, and slowly poured into 30 L of saturated aqueous ammonium chloride. Then the reaction mixture was extracted with ethyl acetate (15 L*3) and the combined organic phase was washed with saturated brine (20 L*2), dried with anhydrous Na2SO4, and filtered. After the solvent was distilled off under reduced pressure, the residue was suspended in n-heptane (4 L) and the solid was filtered off and suspended in ethyl acetate (5 L) to give the compound of Formula V as a yellow-yellow solid. white (1.53 kg, 4.55 mol, 69.7%). 1H NMR (400 MHz, DMSO-d6) δ 7.49 - 7.54 (m, 1 H), 6.54 - 6.62 (m, 2 H), 6.15 - 6.20 (m, 1 H), 5.54 (s, 2 H), 4.10 - 4.22 (m, 2 H), 3.42 - 3.55 (m, 2 H), 1.58 - 1.73 (m , 2 H), 1.35 - 1.47 (m, 2 H), 0.90 - 0.96 (m, 3 H), 0.83 - 0.89 (m, 2 H), 0.05 (s, 9 H). Formula VI: 2-butoxy-5H-pyrrolo[3,2-d]pyrimidin-4-amine

[0085] The compound of Formula V (1.10 kg, 3.27 mol) was dissolved in TFA (5.50 L) and the reaction liquid was stirred at 25°C for 16 hours. The reaction was monitored by HPLC. Upon completion, TFA was removed by distillation under reduced pressure and the residue was dissolved in methanol (1.2 L) and ice water (1.2 L). pH of the system was adjusted to 12 with concentrated aqueous ammonia under uniform stirring. The mixture was stirred for 2 hours and the precipitate was precipitated from the solution continuously. After filtration, the filtered mass as a white solid was suspended with 15% aqueous ammonia (1.2 L*3) and ethyl acetate (4 L) successively to yield the compound of Formula VI as a white solid (550.00 g, 2.67 mol, 81.7%). 1H NMR (400 MHz, methanol-d4) δ 7.37 (d, J=2.89 Hz, 1 H), 6.29 (d, J=3.01 Hz, 1 H), 4.27 (t , J=6.53 Hz, 2 H), 1.75 (d, J=7.91 Hz, 2 H), 1.44 - 1.61 (m, 2 H), 1.00 (t, J =7.40 Hz, 3 H). Formula VII: 4-((4-amino-2-butoxy-5H-pyrrolo[3,2-d]pyrimidin-7-yl)-hydroxymethyl)benzaldehyde

[0086] In a three-neck flask, terephthalaldehyde (790.64 mg, 5.82 mmols) and isopropanol (10 mL), 2-butoxy-5H-pyrrolo[3,2-d]pyrimidin-4-amine ( 1.00 g, 4.85 mmols) was added with stirring, and the system was cooled to 0°C and stirred for another 10 minutes. Purified water (10 mL) and potassium carbonate (804.17 mg, 582 mmol) were added, and reacted at 25°C for 16 hours until the reagents were flushed with the monitor by LCMS. A solid was precipitated after the reaction was complete. After filtration the resulting solid was suspended with 20 mL of purified water and 30 mL (ethyl acetate/n-heptane=1/20) successively, filtered and dried to yield the compound of Formula VII as a yellow solid (1.50 g, 4.41 mmol, yield: 90.9%). 1H NMR (400 MHz, methanol-d4) δ 9.94 (s, 1 H), 7.86 (d, J = 8.16 Hz, 2 H), 7.72 (d, J = 8.16 Hz , 2 H), 7.12 - 7.17 (m, 1 H), 6.19 (s, 1 H), 4.28 (t, J=6.53 Hz, 2 H), 1.68 - 1.77 (m, 2 H), 1.44 - 1.54 (m, 2 H), 0.97 (t, J=7.34 Hz, 3 H). Formula VIII: (4-amino-2-butoxy-5H-pyrrolo[3,2-d]pyrimidin-7-yl)(4-(pyrrolidin-1-ylmethyl)phenyl) methanol

[0087] In a 30 L reactor, the compound of Formula VII (450.0 g, 1.32 mol) and isopropanol (4.5 L) were added, and the mixture was stirred for 5 minutes. Then glacial acetic acid (119.0 g, 1.98 mol) was added and the temperature decreased to 0-10°C with stirring. Pyrrolidine (112.4 g, 1.58 mol) was added dropwise, with the temperature below 10°C. After addition, sodium triacetoxyborohydride (420.0 g, 1.98 mol) was added in portions and reacted at 10-20°C for 3 hours until the raw materials were emptied with the liquid chromatography monitor. After completion of the reaction, 5 L of purified water was added and the solution temperature was lowered to about -10°C, and 12 L of 15% aqueous ammonia was added to the solution, with the solution temperature below 0 °C during addition. Solid was precipitated under stirring. Filtration was performed and the resulting filtered mass was suspended with 2 L of water and 2 L*2 of ethyl acetate. Filtration was performed and drying was conducted under reduced pressure at 40°C for 12 hours to produce the compound of Formula VIII as a yellow solid (465.0 g, 1.18 mol, yield 89.4%, moisture 0.9%) . 1H NMR (400 MHz, methanol-d4) δ 7.46 (d, J=7.91 Hz, 1 H), 7.29 (d, J = 8.03 Hz, 1 H), 7.09 (s , 1 H), 6.12 (s, 1 H), 4.29 (t, J = 6.53 Hz, 2 H), 3.60 (s, 2 H), 2.52 (br, s, , 4 H), 1.66 - 1.83 (m, 6 H), 1.49 (d, J = 7.53 Hz, 2 H), 0.98 (t, J = 7.40 Hz, 3 H). Formula I: 2-butoxy-7-(4-(pyrrolidin-1-ylmethyl)benzyl)-5H-pyrrolo[3,2-d]pyrimidin-4-amine

[0088] The compound of Formula VIII (440.0 g, 1.11 mol) and dichloromethane (7.0 L) were added to the 20L flask, and the system temperature was reduced to below -15°C with stirring. After triethylsilane (880 mL, 5.55 mol) was added dropwise to the system, trifluoroacetic acid (880 mL) was added dropwise continuously, with the temperature maintained below -10°C during addition. After addition, the reaction was carried out at 0°C for 2 hours and monitored by liquid chromatography until the raw material spot disappeared. After completion of the reaction, the reaction liquid was concentrated to dryness, and 2.2 L of ethyl acetate was added to the solution. Stirring was carried out to decrease the temperature to below 0°C. Then saturated sodium carbonate solution was added to adjust the solution to pH 9-10 during which the system temperature was maintained below 10°C. Filtration was performed and the resulting filtered mass was suspended with 2.2 L of water. Filtration was carried out and drying was conducted under reduced pressure to produce 550 g of trifluoroacetate of the compound of Formula I as a white solid.

[0089] In 1.6 L of ethanol, 525g of trifluoroacetate of the compound of Formula I was added as a white solid, and the temperature of the system was reduced to about 0°C with stirring. Then 2.2 L of 1 mol/L sodium hydroxide solution was added. Filtration was performed and the resulting filtered mass was suspended with 2.5 L of purified water. Filtration was performed and drying was conducted under reduced pressure to produce 380.0 g of the compound of Formula I as a solid. 1H NMR (400 MHz, methanol-d4) δ 7.27 (d, J = 8.0 Hz, 2H), 7.22 (d, J = 8.0 Hz, 2H), 7.04 (s, 1H ), 4.32 (t, J = 6.6 Hz, 2H), 3.99 (s, 2H), 3.60 (s, 2H), 2.55 - 2.52 (m, 4H), 1 .85 - 1.71 (m, 6H), 1.55-1.48 (m, 2H), 1.00 (t, J = 7.4 Hz, 3H).

Example 1: Preparation of crystalline form C of 2-butoxy-7-(4-(pyrrolidin-1-ylmethyl)benzyl)-5H-pyrrolo[3,2-d]pyrimidin-4-amine maleate

[0090] The compound of Formula I (3.80 g, 10 mmol) was added to 10 mL of ethanol and heated to dissolve. Then, maleic acid (2.55 g, 22 mmol) was added to the solution and stirring was carried out with continuous heating until the solution became clear. The solution was then allowed to cool to room temperature and stand for 1-2 h. The precipitate was filtered and the solid was dried under reduced pressure with an oil pump to yield maleate crystalline form C of the compound of Formula I as a solid. In the resulting maleate, the molar ratio of the compound of Formula I to maleic acid was 1:2.

[0091] XRPD was measured as follows: device: Bruker D8 ADVANCED X-ray diffractometer; method: target: Cu: K-alpha; wavelength À=1.54179Â; voltage: 40 kV; current: 40 MA; scanning range: 4~40°; sample rotation speed: 15 rpm; Scan speed: 10°/minutes.

[0092] The crystal compound obtained had diffraction peaks substantially as shown in Figure 1.

Example 2: Preparation of crystal form D of 2-butoxy-7-(4-(pyrrolidin-1-ylmethyl)benzyl)-5H-pyrrolo[3,2-d]pyrimidin-4-amine maleate

[0093] In 0.8 ml of acetone, crystalline form C of compound maleate of Formula I (50 mg) was added to form a suspension. The suspension sample was stirred in a thermostat (40°C) for 2 days (in the dark). The residual solid was centrifuged and dried in a vacuum oven at 40 ° C overnight to produce maleate crystalline form D of the compound of Formula I as a solid. In the resulting maleate, the molar ratio of the compound of Formula I to maleic acid was 1:2.

[0094] XRPD was measured as follows: apparatus: Bruker D8 ADVANCED X-ray diffractometer; method: target: Cu: K-alpha; wavelength À=1.54179Â; voltage: 40 kV; current: 40 MA; scanning range: 4~40°; sample rotation speed: 15 rpm; Scan speed: 10°/minutes.

[0095] The crystal compound obtained had diffraction peaks substantially as shown in Figure 2.

Example 3: Preparation of crystalline form E of 2-butoxy-7-(4-(pyrrolidin-1-ylmethyl)benzyl)-5H-pyrrolo[3,2-d]pyrimidin-4-amine maleate

[0096] In 0.8 ml of isopropanol, crystalline form C of compound maleate of Formula I (50 mg) was added to form a suspension. The suspension sample was stirred in a thermostat (40°C) for 2 days (in the dark). The residual solid was centrifuged and dried in a vacuum oven at 40 ° C overnight to produce maleate crystalline form E of the compound of Formula I as a solid. In the resulting maleate, the molar ratio of the compound of Formula I to maleic acid was 1:2.

[0097] XRPD was measured as follows: device: Bruker D8 ADVANCED X-ray diffractometer; method: target: Cu: K-alpha; wavelength À=1.54179Â; voltage: 40 kV; current: 40 MA; scanning range: 4~40°; sample rotation speed: 15 rpm; Scan speed: 10°/minutes.

[0098] The crystal compound obtained had diffraction peaks substantially as shown in Figure 3.

Example 4: High Temperature Stability Test

[0099] The crystalline forms prepared in Examples 1-3 were tested in the accelerated test under high temperature conditions for stability in accordance with Guidelines for the Stability Test of Pharmaceutical Ingredients and Pharmaceutical Preparations (Chinese Pharmacopoeia 2010 Appendix XIXC).

[00100] Crystalline forms C, D and E prepared in Examples 13 were placed in a clean, open container at 60°C, respectively. Samples were taken for testing on day 10, day 20 and day 30 respectively. The results were compared to the initial test result on day 0.

Example 5: High Humidity Stability Test

[00101] The crystalline forms prepared in Examples 1-3 were tested in the accelerated test under high humidity conditions for stability in accordance with Guidelines for the Stability Test of Pharmaceutical Ingredients and Pharmaceutical Preparations (Chinese Pharmacopoeia 2010 Appendix XIXC).

[00102] Crystalline forms C, D and E prepared in Examples 13 in an accelerated test in a constant temperature and humidity container under conditions of 40°C/75% humidity (open). Samples were taken for testing on day 30, day 60 and day 90 respectively. The results were compared with the initial test result on day 0.

Example 6: Light Stability

[00103] The crystalline forms prepared in Examples 1-3 were tested under light conditions for stability in accordance with Guidelines for the Stability Test of Pharmaceutical Ingredients and Pharmaceutical Preparations (Chinese Pharmacopoeia 2010 Appendix XIXC).

[00104] Crystalline forms C, D and E prepared in Examples 13 were respectively placed in an illumination environment of 5000 Lx ± 500 Lx. Samples were taken on day 5, day 10 and day 30. Test results were compared with the initial test result on day 0. Examples of Pharmacological Activity Efficacy Example 1: Toll-like receptor 7 and Toll-like receptor 8 in vitro receptor binding activity sieve Reagents: HEK-blue hTLR7 cell and HEK-blue hTLR8 cell (available from InvivoGen) DMEM medium Heat-infected fetal bovine serum reagent Anti Mycoplasma NormocinTM bleomycin blasticidin

[00105] The structure of GS-9620 and R848 used is as follows, in which the preparation of GS-9620 could refer to the process described in US20100143301; R848 was commercially available from ABGENT (IMG-2208, specification: 0.5 mg).

Scheme: 1. Preparation of 96-well composite plate:

[00106] Compounds were gradient diluted with DMSO by 3 times using liquid workstation POD starting at a concentration of 10 mmol/L and 10 points were diluted (2nd column to 11th column, and each point was doubled). To the 12th column, i μL of 5 mg/mL positive compound R848 was added as a positive control; and the 1st column, 1 μL of DMSO was added as a negative control. Each well contained 1 μL of DMSO. 2. Cells in culture flask were collected and cell density was diluted to 250,000 cells/mL. 3. 200 μL (50,000 cells/well) of cell suspension was added into prepared compound plate and the final concentration of DMSO in each well was 0.5%. 4. The culture plates containing cells and compounds were incubated in a CO2 incubator for 24 hours at 37°C, 5% CO2. 5. After 24 h of incubation, 20 μL of supernatant was removed from each well in a 96-well transparent assay plate. To each well of the assay plate, 180 μL of Quanti-Blue reagent was added and the plate was incubated in an incubator at 37°C, 5% CO2 for 1 hour. 6. After 1 h, the alkaline phosphatase content in 20 μL of supernatant was determined using Microplate Reader OD650. 7. EC50 of each compound was obtained with Prism software.

[00107] Results were shown in Table 1: Table 1

[00108] As shown in table 1, the compound of Formula I according to the invention showed higher in vitro receptor binding activity to Toll-like receptor 7 than the control (Toll-like receptor 7 agonist GS-9620 ) and lower in vitro receptor binding activity to Toll-like receptor 8 than control (Toll-like receptor 7 agonist GS-9620).

[00109] The compound of the present invention has distinct selectivity differences with respect to different receptors, and the effect is superior over the prior art.

Efficacy Example 2: Peripheral Blood Mononuclear Cell Testing Scheme

[00110] The purpose of this example is to determine the level of cytokine expression 24 h after stimulating human peripheral blood mononuclear cells (PBMC) with the compound of Formula I.

[00111] Cell supernatant was analyzed without dilution and IFN-α levels were directly determined. The compound of Formula I was first formulated in 20 mM DMSO raw material solution and was gradient diluted with cell medium by 10 times with the total number of 11 dilution points. Compounds at 9 dilution points (the highest concentration was 200 μmol/L) were added in a 96-well plate with 50 μL in each well. Fresh human peripheral blood mononuclear cells were inoculated, with 150 μL in each well containing 450,000 cells. The cell culture plate was incubated in an incubator at 37°C, 5% CO2 for 24 hours. After incubation, the culture plate was centrifuged at 1200 rpm for 5 minutes and the supernatant was collected and stored at −20°C for determination. Cytokine determination was performed using a BD-Pharmingen Cytometric Bead Array (CBA) on a flow cytometer. Using the previous determination method, the lowest drug concentration stimulating the production of 30 pg/mL of IFN-α was designated as the MEC value in the cytokine stimulation test. The results were shown in Table 2. Table 2

[00112] Conclusion: Compared with the control (GS-9620), the compound of Formula I of the invention showed better in vitro INF-α induction activity of PBMCs and comparable TNF-α induction activity.

Claims (14)

1. Maleate, characterized by the fact that it is a compound of Formula I, 2. Maleate, according to claim 1, characterized by the fact that the moral proportion of the compound of Formula I to maleic acid is 1:1 to 1:3, or 1:2. 3. Crystalline form C of the maleate of the compound of Formula I, characterized by the fact that it presents diffraction peaks at 2θ=7.6°±0.2°, 9.9°±0.2°, 17.8°± 0.2°, 22.8°±0.2°, 24.2°±0.2°, 26.3°±0.2° in X-ray powder diffraction pattern, 4. Crystalline form C, according to claim 3, characterized by the fact that it presents diffraction peaks at 2θ=5.6°±0.2°, 7.6°±0.2°, 9.9°± 0.2°, 17.8°±0.2°, 19.8°±0.2°, 22.8°±0.2°, 24.2°±0.2°, 25.0°± 0.2°, 26.3°±0.2° in X-ray powder diffraction pattern; for example, it presents diffraction peaks at 2θ=5.6°±0.2°, 6.0°±0.2°, 7.6°±0.2°, 9.9°±0.2°, 12.0°±0.2°, 15.3°±0.2°, 17.8°±0.2°, 18.5°±0.2°, 19.8°±0.2°, 20, 4°±0.2°, 22.8°±0.2°, 23.1°±0.2°, 24.2°±0.2°, 24.7°±0.2°, 25, 0°±0.2°, 26.3°±0.2°in X-ray powder diffraction pattern; for example, it presents an X-ray powder diffraction pattern substantially depicted in Figure 1. 5. Process for preparing crystalline form C, as defined in claim 3 or 4, characterized by the fact that it comprises the following steps: (1) dissolving the compound of Formula I in a solvent, which is heated to promote dissolution; (2) add maleic acid; and (3) cool for crystallization, filter, wash and dry to obtain crystalline form C, with the solvent being ethanol. 6. Crystalline form D of the maleate of the compound of Formula I, characterized by the fact that it presents diffraction peaks at 2θ=5.1°±0.2°, 9.5°±0.2°, 11.2°± 0.2°, 17.6°±0.2°, 20.2°±0.2°, 23.0°±0.2° in X-ray powder diffraction pattern, 7. Crystalline form D, according to claim 6, characterized by the fact that it presents diffraction peaks at 2θ=5.1°±0.2°, 9.5°±0.2°, 11.2°± 0.2°, 17.6°±0.2°, 20.2°±0.2°, 20.7°±0.2°, 23.0°±0.2°, 23.7°±0, 2° in the X-ray powder diffraction pattern; for example, it presents diffraction peaks at 2θ=5.1°±0.2°, 5.6°±0.2°, 9.5°±0.2°, 11.2°±0.2°, 16.9°±0.2°, 17.6°±0.2°, 20, 2°±0.2°, 20.8°±0.2°, 22.6°±0.2°, 23.0°±0.2°, 23.7°±0.2°, 24.5° ±0.2°in X-ray powder diffraction pattern; for example, it presents an X-ray powder diffraction pattern substantially depicted in Figure 2. 8. Process for preparing crystalline form D, as defined in claim 6 or 7, characterized by the fact that it comprises the following steps: (1) placing the crystalline form C of the maleate of the compound of Formula I, as defined in claim 3 or 4, in acetone solvent to form a suspension; (2) stir at constant temperature; and (3) centrifuge, wash, and dry to obtain crystalline form D. 9. Crystalline form E of the maleate of the compound of Formula I, characterized by the fact that it presents diffraction peaks at 2θ=4.9°±0.2°, 5.3°±0.2°, 9.0°± 0.2°, 16.5°±0.2°, 19.3°±0.2° in X-ray powder diffraction pattern, 10. Crystalline form E, according to claim 9, characterized by the fact that it presents diffraction peaks at 2θ=4.9°±0.2°, 5.3°±0.2°, 6.7°± 0.2°, 9.0°±0.2°, 10.8°±0.2°, 16.5°±0.2°, 19.3°±0.2°in powder diffraction pattern X-ray; for example, crystal form E exhibits diffraction peaks at 2θ=4.9°±0.2°, 5.3°±0.2°, 6.7°±0.2°, 9.0°±0.2°, 10.8°±0.2°, 16.2°±0.2° , 16.5°±0.2°, 19.3°±0.2°, 22.0°±0.2°, 22.6°±0.2°, 25.9°±0.2° X-ray powder diffraction; for example, crystalline form E exhibits an X-ray powder diffraction pattern substantially depicted in Figure 3. 11. Process for preparing crystalline form E, as defined in claim 9 or 10, characterized by the fact that it comprises the following steps: (1) placing the crystalline form C of the maleate of the compound of Formula I, as defined in claim 3 or 4, in isopropanol solvent to form a suspension; (2) stir at constant temperature; and (3) centrifuge, wash and dry to obtain crystalline form E. 12. Crystalline composition, characterized by the fact that: based on the weight of the crystalline composition, the crystalline form C, as defined in claim 3 or 4, is 50% or more, 80% or more, 90% or more, or 95 % or more; or based on the weight of the crystalline composition, the crystalline form D, as defined in claim 6 or 7, is 50% or more, 80% or more, 90% or more, or 95% or more; or based on the weight of the crystalline composition, the crystalline form E as defined in claim 9 or 10 is 50% or more, 80% or more, 90% or more, or 95% or more. 13. Pharmaceutical composition, characterized by the fact that it comprises: the maleate of the compound of Formula I, as defined in claim 1 or 2, the crystalline form C, as defined in claim 3 or 4, the crystalline form D, as defined in claim 6 or 7, the crystalline form E as defined in claim 9 or 10, or the crystalline composition as defined in claim 12, or any combination thereof in an effective amount; and one or more pharmaceutically acceptable carriers and/or excipients. 14. Use of the maleate of the compound of Formula I, as defined in claim 1 or 2, of crystalline form C, as defined in claim 3 or 4, of crystalline form D, as defined in claim 6 or 7, of crystalline form E, as defined in claim 9 or 10, of the crystalline composition, as defined in claim 12, or of the pharmaceutical composition, as defined in claim 13, said use being characterized by the fact that it is for manufacturing a medicament to treat disease associated with the receptor 7 similar to Toll, selected from infection with the hepatitis B or hepatitis C virus.