Process for preparation of ridinilazole and crystalline forms thereof

Abstract

To provide a mixture comprising ridinilazole, which is a potent C.difficile-targeting antimicrobial.SOLUTION: The present invention provides a composition comprising a mixture of compounds, the mixture comprising ridinilazole and impurities E represented by formula (II) and impurities F represented by formula (IV), where the combined amount of impurities E and F in the mixture is less than 100 ppm.SELECTED DRAWING: Figure 1

Description

Technical Field

[0001] 1. Field of the Invention The present invention relates to 2,2'-di(pyridin-4-yl)-1H,1'H-5,5'-bibenzene [d]imidazole (which may also be known as 5,5'-bis[2-(4-pyridinyl)-1H-benzimidazole], 2,2'-bis(4-pyridyl)-3H,3'H-5,5'-bibenzimidazole or 2-pyridin-4-yl-6-(2-pyridin-4-yl-3H-benzimidazole-5-yl)-1H-benzimidazole), which is referred to herein by the INN name ridinilazole, and methods for the preparation of pharmaceutically acceptable derivatives, salts, hydrates, solvates, complexes, biological equivalents, metabolites or prodrugs thereof. The present invention relates to various crystalline forms (crystal forms) of ridinilazole, methods for their preparation, and related pharmaceuticals and their use (including their medical use and use in the efficient large-scale synthesis of ridinilazole).

Background Art

[0002] 2. Background of the Invention Infection with Clostridioides difficile (formerly known as Clostridium difficile) (CDI) causes Clostridioides difficile-associated diseases (CDAD). In the United States, more than 450,000 cases of CDI occur each year, more than 80,000 are first recurrences, and approximately 29,000 people die. The most common factor is the use of antibiotics ​​​​​​​​​is used. Antibiotics can potentially lead to the formation of a less diverse microbiota that is more susceptible to pathogen invasion over a long period of time, causing a loss of colony-forming resistance. Oral vancomycin and oral metronidazole treatment is associated with a high recurrence rate of CDI and is likely to have an adverse effect on the resident colonic microbiota. Recurrence is costly from both the perspectives of clinical burden and utilization of medical resources. In one study, readmission was necessary in approximately one-third of the recurrent cases. Both the biomass of the gut microbiota and the composition at the gut-bacteria interface may influence the niche for Clostridioides difficile colony formation. Colony-forming resistance is associated with specific taxa but is thought to be able to provide protection to different and diverse microbial community structures. Consistent characteristics of communities susceptible to CDI are low levels of diversity, reduced metabolic function, and a relative decrease in members of the phyla Bacteroidetes and Firmicutes and an increase in members of Proteobacteria. Fecal microbiota transplantation (FMT) normalizes these characteristics and breaks the CDI recurrence cycle. Overall, these data support the role of CDI agents in reducing the risk of recurrence while minimizing the impact on the resident microbiota. Rizinilazole (also known as SMT19969, 2,2’-di(pyridin-4-yl)-1H,1’H-5,5’-bibenzimidazole or 5,5’-bis[2

[0003] intestinal bacteria biomass and the composition of the composition at the intestinal and bacterial interface both potentially affect the niche for the formation of C. difficile colonies. Colony-forming resistance is associated with specific taxa but is thought to be able to provide protection to different and diverse microbial community structures. Consistent characteristics of communities susceptible to CDI are low levels of diversity, reduced metabolic function, and a relative decrease in members of the phyla Bacteroidetes and Firmicutes and an increase in members of Proteobacteria. Fecal microbiota transplantation (FMT) normalizes these characteristics and breaks the CDI recurrence cycle. There is a possibility that it affects the niche for the formation of C. difficile colonies. Colony-forming resistance is associated with specific taxa but is thought to be able to provide protection to different and diverse microbial community structures. Consistent characteristics of communities susceptible to CDI are low levels of diversity, reduced metabolic function, and a relative decrease in members of the phyla Bacteroidetes and Firmicutes and an increase in members of Proteobacteria. Fecal microbiota transplantation (FMT) normalizes these characteristics and breaks the CDI recurrence cycle. is associated with specific taxa, but it is thought to be able to provide protection to different and diverse microbial community structures. Consistent characteristics of communities susceptible to CDI are low levels of diversity, reduced metabolic function, and a relative decrease in members of the phyla Bacteroidetes and Firmicutes and an increase in members of Proteobacteria. Fecal microbiota transplantation (FMT) normalizes these characteristics and breaks the CDI recurrence cycle. communities that are susceptible to the effects of CDI have consistent characteristics of low diversity levels, reduced metabolic function, and a relative decrease in members of the phyla Bacteroidetes and Firmicutes and an increase in members of Proteobacteria. Fecal microbiota transplantation (FMT) normalizes these characteristics and breaks the CDI recurrence cycle. are low in diversity levels, have reduced metabolic function, and have a relative decrease in members of the phyla Bacteroidetes and Firmicutes and an increase in members of Proteobacteria. Fecal microbiota transplantation (FMT) normalizes these characteristics and breaks the CDI recurrence cycle. are relatively reduced, and members of Proteobacteria increase. Fecal microbiota transplantation (FMT) normalizes these characteristics and breaks the CDI recurrence cycle. Fecal microbiota transplantation (FMT) normalizes these characteristics and breaks the CDI recurrence cycle. Fecal microbiota transplantation (FMT) normalizes these characteristics and breaks the CDI recurrence cycle.

[0004] Overall, these data support the role of CDI agents in reducing the risk of recurrence while minimizing the impact on the resident microbiota. Fecal microbiota transplantation (FMT) normalizes these characteristics and breaks the CDI recurrence cycle.

[0005] Rizinilazole (also known as SMT19969, 2,2’-di(pyridin-4-yl)-1H,1’H-5,5’-bibenzimidazole or 5,5’-bis[2 pyridin-4-yl)-1H,1’H-5,5’-bibenzimidazole or 5,5’-bis[2 -(4-Pyridinyl)-1H-benzimidazole], which can be variously referred to in the literature has a narrow spectrum, low absorbency, and is a potent antibacterial agent targeting Clostridioides difficile. Rizinilazole is represented by the following formula :

[0006]

Chemical formula

[0007] In a recent Phase 2 randomized comparative double-blind clinical trial, rizinilazole significantly reduced the incidence of recurrent disease (14.3% vs. 34. 8%) when compared with vancomycin in terms of efficacy. Rizinilazole shows strong preservation of the human gut microbiota compared to vancomycin (which may contribute to the reduced recurrence of CDI observed in the Phase 2 study).

[0008] Therefore, there is a need for an efficient synthesis of rizinilazole.

[0009] The control of genotoxicity and potentially genotoxic impurities (PGIs) in drug manufacturing is a major concern, and the acceptable levels should not exceed those justified by safety data. There is a need in the technical field for methods that can enable the effective removal of PGIs to prepare drug candidates.

[0010] The present inventors are currently developing methods for efficiently producing rizinilazole and its pharmaceutically acceptable salts, hydrates, solvates, complexes, biological equivalents, metabolites, or prodrugs. These are (a) suitable for large-scale synthesis under GMP conditions and (b) reduce PGIs to an acceptable level for commercial manufacture of formulations. ​

[0011] The inventors have now also discovered three different crystalline forms (polymorphs) of ridinilazole which are particularly useful in the above methods and have found applications in the efficient large-scale synthesis of ridinilazole for medical use (and generally in the medical field).

[0012] 3. Prior Art WO 2010 / 063996 pamphlet describes various benzimidazoles containing ridinilazole and their use as antibacterial agents (including the treatment of CDAD).

[0013] WO 2011 / 151621 pamphlet describes various benzimidazoles and their use as antibacterial agents (including the treatment of CDAD).

[0014] WO 2007056330 pamphlet, WO 2003105846 pamphlet and WO 2002060879 pamphlet disclose various 2-aminobenzimidazoles as antibacterial agents.

[0015] WO 2007148093 pamphlet discloses various 2-aminobenzothiazoles as antibacterial agents.

[0016] WO 2006076009 pamphlet, WO 200 / 041209 pamphlet, and Bowser et al. (Bioorg. Med. Chem. Lett., 2007, 17, 5652-5655) disclose various substituted benzenes useful as anti-infective agents that reduce the resistance, pathogenicity, or growth of microorganisms. A zuidimidazole compound is disclosed. This compound is said not to exhibit endogenous antibacterial activity in vitro.

[0017] U.S. Patent No. 5,824,698 discloses various dibenzimidazoles as broad-spectrum antibiotics, showing activity against both Gram-negative and Gram-positive bacteria, including the genera Staphylococcus and Enterococcus. However, this document does not disclose activity against anaerobic spore-forming bacteria, and in particular, does not disclose activity against any bacteria of the genus Clostridioides (including Clostridioides difficile).

[0018] U.S. Patent Application Publication No. 2007 / 0112048 discloses various biarylimidazolidine, triarylimidazolidine, biarylamidine, and triarylamidine as broad-spectrum antibiotics, showing activity against both Gram-negative and Gram-positive bacteria, including the genera Staphylococcus, Enterococcus, and Clostridioides. However, this document does not disclose the compounds of formula (I) described herein.

[0019] Chaudhuri et al. (2007) J. Org. Chem. 72, 1912-1923 discloses various bi- S-2-(pyridyl)-1H-benzimidazole (including the compound of formula I described in this specification) is described. This document is silent about potential antibacterial activity.

[0020] Singh et al. (2000) Synthesis 10: 1380-1390 describes a condensation reaction for producing 2,2'-di(pyridin-4-yl)-1H,1'H-5,5'-bibenzimidazole using 4-pyridinecarboxaldehyde, FeCl3, and O2 in DMF at 120 °C. -yl)-1H,1'H-5,5'-bibenzimidazole is described.

[0021] Bhattacharya and Chaudhuri (2007) Chemistry - An Asian Journal 2: 648-655 describes a condensation reaction for producing 2,2'-di(pyridin-4-yl)-1H,1'H-5,5'-bibenzimidazole using 4-pyridinecarboxaldehyde and nitrobenzene at 120 °C. -di(pyridin-4-yl)-1H,1'H-5,5'-bibenzimidazole is described.

[0022] WO 2019 / 068383 describes the synthesis of lysinilazole by coupling 3,4,3',4'-tetraaminobiphenyl and 4-pyridinecarboxaldehyde with a metal ion catalyst in the presence of oxygen and then adding a complexing agent. -pyridinecarboxaldehyde with a metal ion catalyst and then adding a complexing agent. SUMMARY OF THE INVENTION

[0023] 4. Summary of the Invention According to a first aspect of the present invention, there is provided a composition comprising a mixture of compounds, wherein the mixture comprises lysinilazole and formulas (II) and (IV):

[0024]

Chemical formula

[0025] In a preferred embodiment, the lysinilazole is in the crystalline form (Form A) of lysinilazole tetrahydrate characterized by powder X-ray diffraction (XRPD) patterns having peaks characteristic of 2 theta angles of (11.02 ± 0.2)°, (16. 53 ± 0.2)° and (13.0 ± 0.2)°. (Form A). exists as

[0026] In a second aspect of the present invention, there is provided a method for producing the composition according to the first aspect of the present invention, comprising: (a) providing a crude lysinilazole composition comprising a mixture of compounds, said mixture comprising lysinilazole and compounds of formula (II) and formula (IV): wherein the total amount of impurity E and impurity F in the mixture exceeds 100 ppm;

[0027]

Chemical formula

[0028] In a third aspect, the present invention provides a composition according to the first aspect of the present invention obtainable (or produced) by the method of the present invention. is provided.

[0029] In another aspect, the present invention provides a pharmaceutical composition comprising an effective amount of the composition of the present invention and a pharmaceutically acceptable excipient. Provided is a pharmaceutical composition comprising an agent.

[0030] In another aspect, the present invention provides a composition of the present invention for use in treatment or prevention. Thereof.

[0031] In another aspect, the present invention provides a composition of the present invention for use in the treatment or prevention of CDI or CDAD. Thereof.

[0032] In another aspect, the present invention provides the use of a composition of the present invention for the manufacture of a medicament for the treatment, therapy or prevention of CDI or CDAD. Thereof.

[0033] In another aspect, the present invention provides a crystalline form (Form A) of lirinizole tetrahydrate characterized by peaks at 2 theta angles of (11.02 ± 0.2)°, (16.53 ± 0.2)° and (13.0 ± 0.2)° in a powder X-ray diffraction pattern. Thereof.

[0034] In another aspect, the present invention provides a crystalline form (Form D) of lirinizole anhydrate characterized by peaks at 2 theta angles of (12.7 ± 0.2)°, (23.18 ± 0.2)° and (27.82 ± 0.2)°, optionally including peaks at 2 theta angles of (12. 7 ± 0.2)°, (23.18 ± 0.2)°, (27.82 ± 0.2)°, (19.5 ± 0.2)° and (22.22 ± 0.2)° in a powder X-ray diffraction pattern. Thereof.

[0035] Other aspects and embodiments of the present invention are set forth in the claims appended hereto. Thereof. BRIEF DESCRIPTION OF THE DRAWINGS

[0036] 6. BRIEF DESCRIPTION OF THE DRAWINGS

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Figure 17

Figure 18

Figure 19

MODE FOR CARRYING OUT THE INVENTION

[0037] 5. DETAILED DESCRIPTION AND EXAMPLES OF THE INVENTION All publications, patents, patent applications and other references described herein are incorporated by reference in their entirety for all purposes as if each individual publication, patent or patent application was specifically and individually indicated to be incorporated by reference and the content thereof was fully recited. and incorporated herein by reference in their entirety as if each individual publication, patent or patent application was specifically and individually indicated to be incorporated by reference and the content thereof was fully recited. and incorporated herein by reference in their entirety for all purposes as if each individual publication, patent or patent application was specifically and individually indicated to be incorporated by reference and the content thereof was fully recited. and incorporated herein by reference in their entirety for all purposes as if each individual publication, patent or patent application was specifically and individually indicated to be incorporated by reference and the content thereof was fully recited.

[0038] 5.1 Definitions and General Preferences As used herein and unless otherwise indicated elsewhere, the following terms have the following meanings in addition to any broader (narrower) meaning the term may enjoy in the art. As used herein and unless otherwise indicated elsewhere, the following terms have the following meanings in addition to any broader (narrower) meaning the term may enjoy in the art. are shown to have. Unless the context requires otherwise, as used herein, the use of the singular form is read to include the plural form, and vice versa. With respect to terms relating to an entity, the term "a" or "an" is read to refer to one or more of that entity. Thus, as used herein, the terms "a" (or "an"), "one or more", Unless the context requires otherwise, as used herein, the use of the singular form is read to include the plural form, and vice versa. With respect to terms relating to an entity, the term "a" or "an" is read to refer to one or more of that entity. Thus, as used herein, the terms "a" (or "an"), "one or more", Unless the context requires otherwise, as used herein, the use of the singular form is read to include the plural form, and vice versa. With respect to terms relating to an entity, the term "a" or "an" is read to refer to one or more of that entity. Thus, as used herein, the terms "a" (or "an"), "one or more", Unless the context requires otherwise, as used herein, the use of the singular form is read to include the plural form, and vice versa. With respect to terms relating to an entity, the term "a" or "an" is read to refer to one or more of that entity. Thus, as used herein, the terms "a" (or "an"), "one or more", 「At least one」 is used synonymously.

[0039] In this specification, the term "comprise", or variations thereof such as "compr ises" and "comprising", is to be read as indicating the inclusion of any recited component (e.g., feature, element, characteristic, property, method / step of a process, limitation) or group of components (e.g., feature, element, characteristic, property, method / step of a process or limitation ), but not the inclusion of any other component or group of components. Thus, the term "comprising" as used herein is inclusive or open-ended and does not exclude additional, unrecited components or steps of a method / process.

[0040] In this specification, the phrase "consisting essentially of" is used in connection with a particular component or step, as well as those that do not require other components or steps that materially affect the

[0041] characteristics or functions of the claimed invention. When used in this specification, the term "consisting of" is used to indicate the presence of only the recited components (e.g., features, elements, characteristics, properties, steps of a method / process, or limitations), or group of components (e.g., features, elements,

[0042] characteristics, properties, steps of a method / process, or limitations). The pharmaceutical composition of the present invention is included in a pharmaceutical kit, pack, or patient pack.

[0043] As used herein, the term "pharmaceutical kit" defines a pharmaceutical composition in one unit dose or multiple unit doses, arranged together with a dosing means (e.g., a measuring device) and / or a delivery means (e.g., an inhaler or a syringe). The unit dose and the dosing means may optionally all be contained in a common outer package. The unit dose may be contained in a blister pack. The pharmaceutical kit may optionally further include instructions for use. As used herein, the term "pharmaceutical pack" defines the arrangement of a pharmaceutical composition in one unit dose or multiple unit doses, which may optionally be contained in a common outer package. The unit dose may be contained in a blister pack. The pharmaceutical pack may optionally further include instructions for use. As used herein, the term "patient pack" defines a package containing a pharmaceutical composition prescribed for a patient and for the entire course of treatment. A patient pack typically includes one or more blister packs. The patient pack has advantages over conventional prescriptions where a pharmacist divides the supply of a patient's medications from bulk supply, and the patient has access to the accompanying documents that are always included in the patient pack and that are usually lost with the patient's prescription. Inclusion of the accompanying documents has been shown to improve patient compliance with the physician's instructions. As used herein, the term "rilinirazole" refers to the compound 2,2'-di(pyridin-4-yl)-1H,1'H-5,5'-bibenzimidazole (5,5'-

[0044]

[0045]

[0046] ​​​​​​​​​​​​​Bis[2-(4-pyridinyl)-1H-benzimidazole], 2,2'-bis(4-pyr ridyl)-3H,3'H-5,5'-bibenzimidazole or 2-pyridin-4-yl- 6-(2-pyridin-4-yl-3H-benzimidazol-5-yl)-1H-benz imidazole (which may also be known as) is used to define. This term as defined herein, also includes pharmaceutically acceptable derivatives, salts, hydrates, solvates, complexes, biological equivalents, metabolites or prodrugs of riginirazole.

[0047] The term pharmaceutically acceptable derivative as applied to riginirazole defines a compound obtained (or obtainable) by chemical derivatization of the parent compound of the present invention. Thus, a pharmaceutically acceptable derivative is suitable for administration or contact with mammalian tissues (i.e., with a reasonable benefit / risk ratio) without excessive toxicity, irritation or allergic response. Preferred derivatives are those obtained (or obtainable) by alkylation, esterification or acylation of the parent compound of the present invention. The derivative itself may be active or may be inactive until processed in vivo. In the latter case, the derivative of the present invention acts as a prodrug. Particularly preferred prodrugs are ester derivatives esterified with one or more free hydroxyls and activated by hydrolysis in vivo. Other preferred prodrugs are covalent compounds that release the active parent drug of formula (I) after cleavage of the covalent bond in vivo.

[0048] The pharmaceutically acceptable derivatives of the present invention retain some or all of the It exists. In some cases, the activity is increased by derivatization. Derivatization can also enhance other biological activities of the compound, such as bioavailability.

[0049] The term pharmaceutically acceptable salts as applied to riginilazole refers to non-toxic organic addition salts or non-toxic inorganic acid addition salts of any free base compound that are suitable for use in contact with mammalian tissues without undue toxicity, irritation, allergy response, and are commensurate with a reasonable benefit / risk ratio. Suitable pharmaceutically acceptable salts are well known in the art. Examples are salts with inorganic acids ( e.g., hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid), organic carboxylic acids (e.g., acetic acid, propionic acid, glycolic acid, lactic acid, pyruvic acid, malonic acid, succinic acid, fumaric acid, malic acid, tartaric acid, citric acid, ascorbic acid, maleic acid, hydroxymaleic acid, dihydroxymaleic acid, benzoic acid, phenylacetic acid, 4-aminobenzoic acid, 4-hydroxybenzoic acid, anthranilic acid, cinnamic acid, salicylic acid, 2-phenoxybenzoic acid, 2-acetoxybenzoic acid and mandelic acid), and organic sulfonic acids (e.g., methanesulfonic acid and p-toluene sulfonic acid). The compounds of the present invention can be converted to (mono- or di-) salts by reaction with suitable bases, such as, for example, alkali metal hydroxides, methoxides, ethoxides or tert-butoxides or alkyllithiums, such as NaOH, NaOMe, KOH, KOtBu, LiOH and BuLi, selected from and pharmaceutically acceptable salts of riginilazole are thus prepared.

[0050] These salts and the free base compounds can be in either the hydrated form or the substantially anhydrous form. can exist. The crystalline forms of the compounds of the present invention are also contemplated, and in general, the acids of the compounds of the present invention addition salts are soluble in water and various hydrophilic organic solvents and are crystalline materials that demonstrate higher melting points and increased solubility compared to their free base forms. For example, the sodium salt of lysinilazole is sufficiently soluble in methanol such that a methanol solution can pass through / through activated carbon.

[0051] The term pharmaceutically acceptable solvate as applied to lysinilazole refers to any pharmaceutically acceptable solvate of a particular compound that retains the biological effectiveness of such compound. Examples of solvates include the compounds of the invention with water (hydrates), short-chain alcohols (including isopropanol, ethanol, methanol), dimethyl sulfoxide, ethyl acetate, acetic acid, ethanolamine, acetone, dimethylformamide (DMF), dimethyl acetamide (DMAc), pyrrolidones (such as N-methyl-2-pyrrolidone (NMP) ), tetrahydrofuran (THF), and ethers (such as tertiary butyl methyl ether (TBME) ), etc.) in combination. )

[0052] Also included are miscible formulations of solvate mixtures that include the compounds of the present invention together with a mixture of acetone and ethanol. In a preferred embodiment, the solvate includes the compound of the present invention in combination with about 20% ethanol and about 80% acetone. Accordingly, the structural formulas include compounds having the indicated structure, including hydrate and non-hydrate forms.

[0053] The term pharmaceutically acceptable prodrug as applied to lysinilazole refers to a physiologically ​​​​​under the conditions of, or by solvolysis in vivo to, such a pharmaceutically acceptable salt of the compound, or at least a part of the antibacterial activity of the specified compound to a compound that shares (e.g., shows activity against Clostridioides difficile), any pharmaceutically acceptable compound that can be converted is defined. To do.

[0054] The term pharmaceutically acceptable metabolite as applied to ridinilazole defines a pharmacologically active product produced by the metabolism in vivo of ridinilazole or its salts. To define.

[0055] The prodrugs and active metabolites of the compounds of the present invention can be identified using conventional techniques known in the art (e.g., see Bertolini et al., J. Med. Chem., 1997, 40, 2011-2016). For reference). Reference).

[0056] The term pharmaceutically acceptable complex as applied to ridinilazole defines a compound or composition of which the compounds of the present invention form a constituent part. Thus, the complexes of the present invention include derivatives in which the compounds of the present invention are physically associated (e.g., by covalent or non-covalent bonds) with another moiety (one or more). Thus, this term includes the multimeric forms of the compounds of the present invention. Such multimers can be produced by linking or arranging multiple copies of the compounds of the present invention in close proximity to each other (e.g., via a scaffold or carrier moiety). This term includes cyclodextrin complexes. To include. To include. To contact (e.g., via a scaffold or carrier moiety) and link or arrange. To be able to.

[0057] The term biologically equivalent (or simply equivalent) is a term used in the art and , a drug analog in which one or more atoms (or groups of atoms) are replaced with substitution atoms (or groups of atoms) having similar steric and / and / or electronic characteristics is defined. Substituting a hydrogen atom or a hydroxyl group with a fluorine atom is a commonly used biologically equivalent substitution. Sila substitution (C / Si exchange) is a relatively recent technique for producing equivalents. This technique involves the substitution of one or more specific carbon atoms in a compound with silicon (see the article by Tacke and Zilch in Endeavour, New Series, 1986 , 10, 191 - 197 for reconsideration). Sila-substituted equivalents (silicon equivalents) may exhibit improved pharmacological properties, for example, better tolerance, longer half-life, or increased potency (see the article by Englebienne in Med. Chem., 2005, 1(3), 215 - 226). Similarly, substitution of one atom with an isotope, for example, substitution of hydrogen with deuterium, may also lead to improvement in pharmacological properties, for example, it may have a longer half-life (for example, see Kushner et al (1999) Can J Physiol Pharmacol. 77(2): 79 - 88). In its broadest aspect, the invention of the present invention contemplates all biological equivalents (and specifically, all silicon biological equivalents) of the compounds of the present invention. In its broadest aspect, the present invention contemplates all tautomers, optical isomers, racemates, and diastereoisomers of the compounds described herein. Those skilled in the art will recognize that due to the asymmetrically substituted carbon atoms present in the compounds of the present invention, the compounds are optically active and racemic (see the article by Englebienne in Med. Chem., 2005, 1(3), 215 - 226). . Similarly, substitution of one atom with an isotope, for example, substitution of hydrogen with deuterium, may also lead to improvement in pharmacological properties, for example, it may have a longer half-life (for example, see Kushner et al (1999) Can J Physiol Pharmacol. 77(2): 79 - 88). In its broadest aspect, the present invention contemplates all biological equivalents (and specifically, all silicon biological equivalents) of the compounds of the present invention. (for example, see Kushner et al (1999) Can J Physiol Pharmacol. 77(2): 79 - 88). In its broadest aspect, the invention of the present invention contemplates all biological equivalents (and specifically, all silicon biological equivalents) of the compounds of the present invention. In its broadest aspect, the invention of the present invention contemplates all biological equivalents (and specifically, all silicon biological equivalents) of the compounds of the present invention. And specifically, all silicon biological equivalents).

[0058] In its broadest aspect, the present invention contemplates all tautomers, optical isomers, racemates, and diastereoisomers of the compounds described herein. Those skilled in the art will recognize that due to the asymmetrically substituted carbon atoms present in the compounds of the present invention, the compounds are optically active and racemic due to the asymmetrically substituted carbon atoms present in the compounds of the present invention. due to the asymmetrically substituted carbon atoms present in the compounds of the present invention, the compounds are optically active and racemic It will be understood that it can be produced in the form of a mixture. When a chiral center or another isomeric center of another form is present in the compound of the present invention, all forms of such isomers (one or more), including enantiomers and diastereoisomers, are intended to be covered herein. The compounds of the present invention containing a chiral center (or chiral centers) can be used as a racemic mixture, an enantiomer-enriched mixture, or the racemic mixture can be separated using well-known techniques, and the individual enantiomers can be used alone. Accordingly, references to the compounds of the present invention include products as mixtures of diastereoisomers, as individual diastereoisomers, as mixtures of enantiomers, and in the form of individual enantiomers. Accordingly, all forms of such isomers (one or more), including enantiomers and diastereoisomers, are intended to be covered herein. It will be understood that it can be produced in the form of a mixture. When a chiral center or another isomeric center of another form is present in the compound of the present invention, all forms of such isomers (one or more), including enantiomers and diastereoisomers, are intended to be covered herein. The compounds of the present invention containing a chiral center (or chiral centers) can be used as a racemic mixture, an enantiomer-enriched mixture, or the racemic mixture can be separated using well-known techniques, and the individual enantiomers can be used alone. Accordingly, references to the compounds of the present invention include products as mixtures of diastereoisomers, as individual diastereoisomers, as mixtures of enantiomers, and in the form of individual enantiomers. The compounds of the present invention containing a chiral center (or chiral centers) can be used as a racemic mixture, an enantiomer-enriched mixture, or the racemic mixture can be separated using well-known techniques, and the individual enantiomers can be used alone. Accordingly, references to the compounds of the present invention include products as mixtures of diastereoisomers, as individual diastereoisomers, as mixtures of enantiomers, and in the form of individual enantiomers. Accordingly, references to the compounds of the present invention include products as mixtures of diastereoisomers, as individual diastereoisomers, as mixtures of enantiomers, and in the form of individual enantiomers. Accordingly, references to the compounds of the present invention include products as mixtures of diastereoisomers, as individual diastereoisomers, as mixtures of enantiomers, and in the form of individual enantiomers.

[0059] Accordingly, the present invention contemplates all optical isomers of the compounds of the present invention and their racemic forms, and unless otherwise indicated (e.g., by the use of dash-wedge structural formulas), the compounds shown in this specification are intended to include all possible optical isomers of the depicted compounds. Accordingly, the present invention contemplates all optical isomers of the compounds of the present invention and their racemic forms, and unless otherwise indicated (e.g., by the use of dash-wedge structural formulas), the compounds shown in this specification are intended to include all possible optical isomers of the depicted compounds. Accordingly, the present invention contemplates all optical isomers of the compounds of the present invention and their racemic forms, and unless otherwise indicated (e.g., by the use of dash-wedge structural formulas), the compounds shown in this specification are intended to include all possible optical isomers of the depicted compounds. Accordingly, the present invention contemplates all optical isomers of the compounds of the present invention and their racemic forms, and unless otherwise indicated (e.g., by the use of dash-wedge structural formulas), the compounds shown in this specification are intended to include all possible optical isomers of the depicted compounds. If the stereochemical form of the compound is important for pharmaceutical utility, the present invention contemplates the use of the isolated eutomer.

[0060] As used herein, the term condensation reaction refers to a reaction in which two or more reactants produce a single main product, accompanied by the formation of small molecules such as water, ammonia, ethanol, acetic acid, or hydrogen sulfide, as applied to 3,3'-diaminobenzidine (DAB) to produce lysinylazole and the intermediate by-product of formula (II). Accordingly, it is used herein as a term in the broad technical field. As used herein, the term condensation reaction refers to a reaction in which two or more reactants produce a single main product, accompanied by the formation of small molecules such as water, ammonia, ethanol, acetic acid, or hydrogen sulfide, as applied to 3,3'-diaminobenzidine (DAB) to produce lysinylazole and the intermediate by-product of formula (II). As used herein, the term condensation reaction refers to a reaction in which two or more reactants produce a single main product, accompanied by the formation of small molecules such as water, ammonia, ethanol, acetic acid, or hydrogen sulfide, as applied to 3,3'-diaminobenzidine (DAB) to produce lysinylazole and the intermediate by-product of formula (II). Accordingly, it is used herein as a term in the broad technical field. Accordingly, it is used herein as a term in the broad technical field.

[0061] The abbreviation "XRPD" refers to X-ray powder diffraction (or, where the context permits, an X-ray powder diffraction pattern).

[0062] As used herein, the term "room temperature" (RT) relates to a temperature between 15 and 25 °C. .

[0063] The term "substantially identical" with respect to an XRPD diffraction pattern means taking into account variations in peak position and relative peak intensity. The ability to determine substantial identity of X-ray diffraction patterns is within the skill of the art. For example, the normal accuracy of 2-theta values is in the range of ± 0.2° 2-theta. Thus, a diffraction peak that normally appears at 14.9° 2-theta may appear between 14.7° and 15.1° 2-theta on most X-ray diffractometers under standard conditions. Further, variability can also result from the particular apparatus used, as well as the crystallinity, orientation, sample preparation, and other factors of the sample. XRPD measurements are typically carried out at room temperature, for example at a temperature of 20 °C and preferably also at a relative humidity of 40%. . . . . . . .

[0064] As used herein, "Form A" of lizinilazole refers to the crystalline form of lizinilazole tetrahydrate characterized by a powder X-ray diffraction pattern comprising peaks characteristic of 2-theta angles of (11.02 ± 0.2)°, (16.53 ± 0.2)° and (13.0 ± 0.2)°. . . .

[0065] As used herein, "Form N" of lizinilazole comprises peaks characteristic of 2-theta angles of (10.82 ± 0.2)°, (13.35 ± 0.2)° and (19.15 ± 0.2)°, optionally (10.82 ± 0.2)°, (13.35 ± 0.2)°, (19 . . .15 ± 0.2)°, (8.15 ± 0.2)° and (21.74 ± 0.2)° in two theta angles, characterized by a powder X-ray diffraction pattern containing characteristic peaks, refers to the crystalline form of the lysinilazole tetrahydrate .

[0066] As used herein, "Form D" of lysinilazole is at (12.7 ± 0.2)° , (23.18 ± 0.2)° and (27.82 ± 0.2)° in two theta angles, and optionally at (12.7 ± 0.2)°, (23.18 ± 0.2)°, (27.8 2 ± 0.2)°, (19.5 ± 0.2)° and (22.22 ± 0.2)° in two theta angles, characterized by a powder X-ray diffraction pattern containing characteristic peaks, refers to the crystalline form of the lysinilazole anhydrate .

[0067] One skilled in the art will understand that the XRPD pattern can be obtained with measurement errors that depend on the measurement conditions used. In particular, it is generally known that the intensity of the XRPD pattern can vary depending on the measurement conditions used. Since the relative intensity can also vary depending on the experimental conditions, the relative intensity should not be considered decisive. Also, since the measurement error of the diffraction angle of a conventional XRPD pattern is typically about 5% or less, it is necessary to consider the degree of such measurement error when considering the described diffraction angles. It is understood that the various crystalline forms described herein are not limited to crystalline forms in which the resulting X-ray diffraction pattern is exactly the same as the X-ray diffraction pattern shown in the attached figures. Rather, crystalline forms of lysinilazole that exhibit an X-ray diffraction pattern that substantially coincides with the X-ray diffraction pattern shown in the figures (as defined above) fall within the scope of the present invention. It will be understood that the various crystalline forms described herein are not limited to crystalline forms in which the resulting X-ray diffraction pattern is exactly the same as the X-ray diffraction pattern shown in the attached figures. Rather, crystalline forms of lysinilazole that exhibit an X-ray diffraction pattern that substantially coincides with the X-ray diffraction pattern shown in the figures (as defined above) are not limited to crystalline forms in which the resulting X-ray diffraction pattern is exactly the same as the X-ray diffraction pattern shown in the attached figures. Rather, crystalline forms of lysinilazole that exhibit an X-ray diffraction pattern that substantially coincides with the X-ray diffraction pattern shown in the figures (as defined above) fall within the scope of the present invention. ​ .

[0068] As used herein, "substantially" refers to a particular crystalline (polymorphic) form of ridinilazole. The term "essentially pure" means that the compound contains less than 10% by weight of any other physical form of ridinilazole, Preferably, it contains less than 5% by weight, more preferably less than 3% by weight, and most preferably less than 1% by weight. is used to define

[0069] As used herein, the term "impurity E" refers to an impurity of formula (II):

[0070] [ka] means a compound of the formula:

[0071] As used herein, the term "impurity F" refers to an impurity of formula (IV):

[0072] [ka] Define the compound:

[0073] 5.2 Synthesis of crude ridinilazole by imidate-DAB condensation The inventors have used 3,3'-diaminobenzidine (DAB) in a condensation reaction to obtain the lysine By producing nilazole, it is possible to conveniently synthesize a crude ridinilazole composition. In a preferred embodiment, the condensation reaction is carried out by reacting DAB with an imidate. (which may be referred to herein as "imidate-DAB condensation"). The compound preferably has the formula (V):

[0074] [ka] It is a methyl isonicotinimidate.

[0075] In a preferred embodiment, the condensation reaction is as follows: (a) Adding sodium methoxide to 4-cyanopyridine to produce a compound of formula (V) and then (b) Reacting the compound of formula (V) from step (a) with said DAB and includes.

[0076] The imidate-DAB condensation reaction may include two chemical steps: Step 1a: Reaction of 4-cyanopyridine with methanol catalyzed by sodium methoxide to form methylisonicotinic imidate, and Step 1b: Coupling of 3,3'-diaminobenzidine (DAB) with methylisonicotinic imidate to form crude lysinilazole (DAB).

[0077] The condensation reaction (step 1b) can be carried out at a temperature of 10 °C to 160 °C. The reaction can be carried out at the reflux temperature of the solvent at normal pressure (e.g., 152 °C to 154 °C for DMF). The reaction can be carried out in any suitable solvent that does not interfere with the reaction. Suitable solvents include methanol (as shown in the exemplary reaction scheme 1 below). Among others, N-methyl- 2-pyrrolidone (NMP), dimethylformamide (DMF), and dimethylacetamide (DMAc) are included.

[0078] Therefore, the imidate-DAB condensation is (a) Adding sodium methoxide to 4-cyanopyridine in methanol to produce a compound of formula (V) and then (b) Adding the compound of formula (V) from step (a) to a mixture of DAB and acetic acid in methanol or (c) Adding a mixture of DAB and acetic acid in methanol to the compound of formula (V) of step (a) and comprising.

[0079] [Chemical formula]

[0080] In step 1a, by using different combinations of alkoxides / alcohols, other imidates can be produced and used in the condensation reaction. For example, sodium ethoxide / ethanol can be used instead of sodium methoxide / methanol, while on the other hand, other cations (preferably alkali metals such as lithium or potassium) can be substituted for sodium.

[0081] In step 1b, the amount of acetic acid is preferably less than 3.5 equivalents, for example, 2.5 to 3.0 equivalents. Other acids (such as TFA) can also be used instead of acetic acid.

[0082] There is a wide range for manipulating the exact state of the imidate - DAB condensation reaction, and all such manipulations are within the scope of the present invention. Sources that will be useful to those skilled in the art when practicing the present invention include Vogel's Textbook 5 of Practical Organic Chemistry, Fifth Edition, B. S. Furniss et al, Pearson Education Limited, 19 88, which discusses general practical procedures. In addition, the methods of synthesis are described in Comprehensive Heterocyclic Chemistry, Vol. 88. 1 (Eds.: AR Katritzky, CW Rees), Pergamon Press, Oxford, 1984 and Comprehens ive Heterocyclic Chemistry II: A Review of the Literature 1982 - 1995 The Structur e, Reactions,10 Synthesis, and Uses of Heterocyclic Compounds, Alan R. Katritzk y (Editor), Charles W. Rees (Editor), E.F.V. Scriven (Editor), Pergamon Pr, June 1996. are discussed. Other common resources to assist the skilled person include March's Ad vanced Organic Chemistry: Reactions, Mechanisms, and Structure, Wiley - Interscien ce; 5th edition (January 15, 2001).

[0083] The preferred imidate - DAB reaction is schematically shown below.

[0084] [Chemical formula]

[0085] In the exemplary reaction scheme 1 shown above, the condensation reaction starts with a DAB solution in methanol, and when about three - quarters of the imidate is supplied, the reaction mixture becomes a solution in a short time, and then the crude lysinyl azole product precipitates from the solution (and can be recovered as a wet filter cake).

[0086] ​​​The inventors have found that the kinetics of this crystallization method are erratic and, in particular, depend on stochastic nucleation events without wishing to be bound by any theory, it is believed that impurities E and F are entrained within the lysinilazole crystals (and / or within their amorphous regions). For example, during the precipitation process, some unreacted impurity E is believed to be trapped in the product crystals and unable to react further with the imidate (even if a large excess of imidate is present).

[0087] Accordingly, the recovered crude lysinilazole product contains a mixture of impurities E and F together with the anhydrous crystalline form D of lysinilazole characterized by an XRPD pattern substantially in accordance with Figure 3

[0088] 5.3 Impurities E and F in Crude Lysinilazole In the synthesis of crude lysinilazole described in Section 5.2 (above), the reaction of DAB with imidate requires 2 equivalents of imidate to complete the reaction. The inventors have discovered that when DAB reacts with only 1 equivalent of imidate, an intermediate impurity, the compound of formula (II ), herein also referred to as "impurity E", is formed.

[0089]

Chemical formula

[0090] The inventors have also discovered that monoaminobenzidine (MAB, herein referred to as the compound of formula (III) ) is present as an impurity in commercial sources of DAB. The inventors have also found that MAB reacts with imidate to form a second (process) impurity ​​​​​​​It has been discovered that this is a compound of formula (IV) (also referred to herein as "Impurity F") as shown below. The compound was discovered.

[0091]

Chemical formula

[0092] Therefore, the crude lizinilazole produced as described above contains a mixture of compounds, wherein the mixture contains lizinilazole, formula (II), and formula (IV):

[0093]

Chemical formula

[0094] Surprisingly, the inventors have found that despite the use of highly toxic DAB and the formation of impurities E and F ( both compounds of formula (II) and (IV) are potentially genotoxic impurities (PGIs) ), an efficient and large-scale GMP synthesis of lizinilazole suitable for use in the formulation of pharmaceutical compositions for administration at levels for the treatment of CDI and CDA D in humans can be achieved by ensuring that the total amount of impurities E and F is less than 100 ppm. Therefore, the present invention provides a composition comprising a mixture of compounds, wherein the mixture contains lizinilazole and formulas (II) and (IV): It was discovered that it can be achieved by ensuring that the total amount of impurities E and F is less than 100 ppm.

[0095] Therefore, the present invention provides a composition comprising a mixture of compounds, wherein the mixture contains lizinilazole and formulas (II) and (IV): wherein the mixture contains lizinilazole and formulas (II) and (IV):

[0096]

Chemical formula

[0097] In a preferred embodiment, ridinilazole contains characteristic peaks at 2 theta angles of (11.02 ± 0.2)°, (16 .53 ± 0.2)° and (13.0 ± 0.2)°, and is characterized by a powder X-ray diffraction pattern (XRPD) of the crystalline form of ridinilazole tetrahydrate (Form A). (Form A).

[0098] 5.4 Determination of Impurities E and F by HPLC-MS Materials Water, ultra-high quality (such as MilliQ) or equivalent Formic acid, 99% for MS Methanesulfonic acid (MSA), over 99% ultra-pure Methanol, HPLC grade Impurity E Impurity F

[0099] Equipment Balance: Minimum 5-point balance

[0100] System parameters HPLC / MS system: Agilent LC1200, MSD 61508 Column: ACE 3 C18, 100 × 4.6 mm, catalog number ACE-111- 1046 Mobile phase A: 0.1% v / v formic acid aqueous ion-exchange solution Mobile phase B: 0.1% v / v formic acid methanol solution Diluent: 98:2:2 v / v / v (water: MeOH: methanesulfonic acid) Injection volume: 2 μL Detection: MSD / SIM m / z 302.1 for Impurity E m / z 287.1 for Impurity F Column temperature: 45 °C Flow rate: 1.0 mL / min Autosampler temperature: 5 °C Needle wash: Diluent

[0101] Gradient

[0102]

Table 1

[0103] Post run: 3 minutes

[0104] MSD parameters Spray chamber settings Dry gas flow rate 12.0 L / min Nebulizer pressure 60 psi Dry gas temperature 350 °C Capillary voltage 3000 V

[0105] MSD signal settings Ion source API-ES positive Fragmenter 70 Gain 1.0 SIM ion of impurity E m / z 302.1 at 5.00 minutes SIM ion of impurity F m / z 287.1 at 5.00 minutes

[0106] Injector program

[0107]

Table 2

[0108] Preparation of solution

[0109]

Table 3

[0110] Injection procedure

[0111]

Table 4

[0112] Ingredients: SMT 19969 Impurity E: Retention time is about 6.1 minutes SMT 19969 Impurity F: Retention time is about 6.6 minutes

[0113] System Compatibility The signal-to-noise ratio of each impurity peak in the working standard solution should be less than 10. The retention time window should be within ±1 minute of the predicted retention time of each component listed above.

[0114] 5.5 Removal of Impurities E and F from Crude Riginitol Referring to various dosing regimens indicated for the treatment of CDI or CDAD in human patients By doing so, the inventors have found that the crude riginitol product of the above method contains, in a mixture, the compounds of formula (II) and (IV) (i.e., Impurities E and F respectively) present in such amounts that the total amount is advantageously further purified to less than 100 ppm.

[0115] If the condition that the total amount of Impurities E and F present in the mixture is less than 100 ppm in the purified riginitol composition is satisfied, any suitable purification method or combination of methods can be used.

[0116] Accordingly, the present invention is a method for producing a composition comprising a mixture of compounds, wherein the mixture comprises riginitol and Impurities E and F, and the total amount of Impurities E and F in the mixture is less than 100 ppm, and the method comprises: (a) providing a crude riginitol composition comprising a mixture of compounds, wherein the mixture comprises riginitol and the compounds of formula (II) and (IV):

[0117]

Chemical formula

[0118] The purification method described herein can also be useful for removing or reducing the concentration of other impurities, such as impurities present in starting materials, reactants, and process reagents (such as DAB and MAB), as well as other possible process impurities It will be understood that process impurities

[0119] Preferred purification methods for use as removal step (b) can be used alone or in any combination and are described in more detail below

[0120] 5.5.1 Treatment of Crude Lizinilazole with Imidate to Purge Impurity E The crude lizinilazole product of the imidate-DAB condensation reaction described in Section 5.2 (above) is treated with an imidate solution to react with impurity E, thereby purging it from the mixture

[0121] For example, an imidate solution was prepared using 0.7 equivalents of 4-cyanopyridine, 0.5 equivalents of sodium methoxide, and 7.2 volumes of methanol and stirred at ambient temperature for 2 hours

[0122] To this solution, 5.5 equivalents of acetic acid was added and heated at 40 °C for 30 minutes. The crude lizinilazole produced by the imidate-DAB condensation reaction described in Section 5.2 (above) was added to 9​​​ It was dissolved in 8 volumes of methanol and 4 equivalents of sodium methoxide. The lysinilazole solution was added to the imidate solution at 40 °C over 5 hours and stirred for 10 hours. The mixture was added over 1 hour to ambient temperature and stirred for 1 hour. The slurry was filtered and washed with methanol (2 × 4.5 volumes

[0123] The wet cake was slurried in 12 volumes of methanol at ambient temperature for 2 hours. The slurry was filtered and washed with methanol (2 × 4.5 volumes). The wet cake was dried at 40 °C to give a recovery of 86%.

[0124] The following table summarizes the LCMS results and shows that the imidate treatment is effective in significantly reducing impurity E .

[0125]

Table 5

[0126] Imidate-related impurities introduced by the imidate purge step can be easily removed by carbon treatment (e.g., as described in Section 5.5.6 below). For example, the product from imidate reprocessing is dissolved in MeOH when treated with NaOMe, and the solution is treated with carbon to remove imidate-related impurities, and the lysinilazole product with impurity E purged is precipitated by adding HOAc . . . .

[0127] 5.5.2 Reprecipitation The levels of these associated impurities E and F are reduced by reprecipitating the lysinilazole after dissolving the crude lysinilazole (and thus liberating the associated impurities E and F) . It can be done. This reprecipitation can be conveniently carried out by forming a salt solution (preferably, for example, an alkali metal salt solution in methanol) and then reprecipitating the lysinilazole (for example, by neutralization, for example, by adding acetic acid). After forming the salt solution, the lysinilazole can be reprecipitated (for example, by neutralization, for example, by adding acetic acid). by adding acetic acid).

[0128] Suitable alkali metal salts include sodium salts, potassium salts, and lithium salts.

[0129] After dissolving the crude lysinilazole and sodium methoxide in methanol, it is preferable to precipitate with acetic acid. precipitate with acetic acid).

[0130] Another preferred method is reprecipitation / re-slurrying with DMSO / acetic acid (described in more detail below). described below).

[0131] This reprecipitation step can also be used after the imidate treatment (described in Section 5.5.1 above). above).

[0132] Exemplary reprecipitation process using NaOMe / HOAc The wet cake of the crude lysinilazole produced by the imidate-DAB condensation reaction described in Section 5.2 (above) was analyzed as described herein and found to contain impurity E (17576 ppm) and impurity F (901 ppm). and found to contain impurity E (17576 ppm) and impurity F (901 ppm). ppm).

[0133] The NaOMe / HOAc precipitation (based on 200 g of DAB) can be carried out as follows. 1) Place the wet cake in a reactor. 2) Add methanol to the same reactor (21.5 volumes). 3) Stabilize the temperature of the slurry at 20 - 25 °C. 4) Add a 30% NaOMe / MeOH (4.0 equivalents) solution while maintaining the temperature between 20 - 30 °C Add while maintaining and over a period of at least 30 minutes. 5) Stir the mixture at 20 - 25 °C for at least 30 minutes or until all solids have dissolved. Stir it. 6) Add water (critical injection, 0.9 volume) to the reactor and stir for at least 30 minutes. 7) While maintaining the temperature between 20 - 25 °C, add glacial acetic acid and adjust the pH to 5 - 7 over at least 2 hours. Adjust the pH to 5 - 7. 8) Stir the slurry for at least 6 hours. 9) Filter the slurry. 10) Wash the cake with methanol (18.0 volume) and dry it under vacuum at 40 °C for at least 24 hours. Dry it under vacuum at 40 °C for at least 24 hours.

[0134] By this procedure, the anhydrous crystalline form D of lidiniazole, characterized by an XRPD pattern substantially in agreement with Figure 3, was obtained. Also, the levels of impurities E and F were reduced to 4195 ppm and 303 ppm respectively. By this procedure, the anhydrous crystalline form D of lidiniazole, characterized by an XRPD pattern substantially in agreement with Figure 3, was obtained. Also, the levels of impurities E and F were reduced to 4195 ppm and 303 ppm respectively. Reduced to 4195 ppm and 303 ppm respectively.

[0135] In the above exemplary reprecipitation process, crude lidiniazole is treated with 4 equivalents of sodium methoxide and dissolved in methanol. Lidiniazole has two acidic protons, and theoretically, only 2 equivalents of sodium methoxide are required. Using 2 equivalents of NaOMe instead of 4 equivalents in the above example resulted in better purge efficiency of impurities E and F. Therefore, the reduction of the levels of impurities E and F by the reprecipitation step can be increased by using a stoichiometric amount of the salt - forming agent (here sodium methoxide). In the above exemplary reprecipitation process, crude lidiniazole is treated with 4 equivalents of sodium methoxide and dissolved in methanol. Lidiniazole has two acidic protons, and theoretically, only 2 equivalents of sodium methoxide are required. In the above example, using 2 equivalents of NaOMe instead of 4 equivalents would result in better purge efficiency of impurities E and F. In the above example, using 2 equivalents of NaOMe instead of 4 equivalents would result in better purge efficiency of impurities E and F. Therefore, the reduction of the levels of impurities E and F by the reprecipitation step can be increased by using a stoichiometric amount of the salt - forming agent (here sodium methoxide). Therefore, the reduction of the levels of impurities E and F by the reprecipitation step can be increased by using a stoichiometric amount of the salt - forming agent (here sodium methoxide). Can be increased.

[0136] In the above exemplary reprecipitation process, the reduction of the levels of impurities E and F is achieved by adjusting the pH to 6 - By adding an amount of acetic acid necessary to adjust during 7 (not adding a fixed amount but ) can be further improved.

[0137] Exemplary reprecipitation process using DMSO / HOAc The wet cake of crude manufactured lysinilazole produced by the imidate-DAB condensation reaction described in Section 5.2 (above) was analyzed as described herein and found to contain impurity E (17576 ppm) and impurity F (901 ppm). It turned out.

[0138] 5 g of a sample of this crude lysinilazole composition was slurried with 50 mL of DMSO, and 25 .43 g of HOAc was used to adjust the pH of the mixture from 11.7 to 6.9. The mixture was heated to 100 °C and cooled to ambient temperature.

[0139] By this procedure, the anhydrous crystalline form D of lysinilazole characterized by an XRPD pattern substantially consistent with Figure 3 was obtained. Also, the levels of impurity E and F were reduced to 2 48 ppm and 81 ppm, respectively. It was reduced to.

[0140] 5.5.3 Recrystallization The difference in solubility between lysinilazole and impurities E and F can be utilized in a recrystallization procedure that crystallizes lysinilazole from a solution containing dissolved impurities E and F. This allows lysinilazole to be separated from the dissolved impurities. Containing Lysinilazole can be separated from the dissolved impurities.

[0141] Therefore, removal step (b) may include the step of recrystallizing lysinilazole after dissolving the crude lysinilazole composition in a high-boiling aprotic solvent.

[0142] In a preferred embodiment, the high-boiling aprotic solvent is DMSO.

[0143] In other preferred embodiments, the removal step (b) further comprises slow cooling of the solution and / or temperature cycling. and a cycle.

[0144] Accordingly, the present invention contemplates the use of a recrystallization step of lysinilazole to reduce the level of impurities E and / or F in a composition comprising a mixture of lysinilazole and impurities E and F, wherein the lysinilazole is heated in DMSO such that it enters and then exits the solution.

[0145] The purge effect of this method on impurities E and F can be improved by slow cooling and temperature cycling, and those skilled in the art can easily optimize these parameters by referring to the levels of impurities E and F present in the starting materials (see below) and the lysinilazole mixture.

[0146] Such a recrystallization step can be used after imidate treatment (as described in Section 5.5.1 above).

[0147] Alternatively (or in addition), it can be used after a reprecipitation step (as described in Section 5.5.2 above). For example, it can be used after a step of imidate treatment followed by reprecipitation (see Sections 5.5.1 and 5.5.2 above).

[0148] Exemplary recrystallization method Analysis of the crude lysinilazole product of the imidate-DAB condensation reaction described in Section 5.2 (above) showed that it contained impurity E (474 ppm) and impurity F (65 ppm). ​​​​​​​​​​It was found that. Dry cake (225 g) was added to the reactor, and 20 volumes of DMSO ( 4950 g) and water (112.5 g, 0.5 volume) were added. While stirring the mixture, it was heated to 100 °C.

[0149] Next, the obtained solution was cooled to 25 °C over 2 hours and stirred for at least 2 hours. The obtained slurry was filtered, and the cake was washed with DMSO (990 g, 4 vol) and MTBE (2 × 666 g, 2 × 4 vol). The solid was dried under vacuum at 40 °C for at least 24 hours.

[0150] Analysis of the recovered solid revealed that the levels of impurities E and F were reduced to 5 ppm and 20 ppm, respectively, by the recrystallization process.

[0151] In further experiments, the above procedure was applied to the composition prepared according to Example 12 (below) containing a mixture of lysinilazole hydrate form A and impurities E ( 36 ppm) and impurity F (318 ppm). Analysis of the recovered solid revealed that the levels of impurities E and F were reduced to 3 ppm and 159 ppm, respectively, by the recrystallization process.

[0152] In still further experiments, the above procedure was applied to the composition prepared according to Example 12 (below) containing a mixture of lysinilazole hydrate form A with impurity E added up to ( 2036 ppm) and containing impurity F (318 ppm). Analysis of the recovered solid revealed that the levels of impurities E and F were reduced to 111 ppm and 124 ppm, respectively, by the recrystallization process.

[0153] The purge of impurities can be improved by slow cooling and temperature cycling. Impurity E (up to 2036 pp m) was added, and a composition produced according to Example 12 (below) containing a mixture of impurity F (318 ppm) in the form A of lysinilazole hydrate was used as the starting material. .

[0154] In the experiment of slow cooling, a mixture of this lysinilazole composition and DMSO was heated to 100 °C, held for 4 hours, and then cooled to ambient temperature over 8 hours. It was found that impurities E and F decreased to 306 and 89 ppm, respectively.

[0155] In the temperature cycling experiment, the same mixture was heated to 100 °C, held for 1 hour, then cooled to ambient temperature over 3 hours and held for 1 hour. Next, the mixture was heated to 100 °C over 3 hours, the cooling cycle was repeated 3 times, and then held at ambient temperature for 7 hours. It was found that impurities E and F decreased to 117 and 124 ppm, respectively.

[0156] 5.5.4 Solvent exchange and / or crystallization with lysinilazole alkali metal salts By utilizing the differences in solubility of alkali metal salts of lysinilazole (such as sodium, lithium, potassium salts, etc.) in various solvents, the trapped impurities E and F can be removed. For example, by utilizing the differences in solubility of sodium lysinilazole in various solvents, the trapped impurities E and F can be removed.

[0157] Therefore, the present invention contemplates the use of a sodium lysinilazole solvent exchange step to reduce the levels of impurities E and / or F. In this step, sodium lysinilazole mixed with impurities E and F in a first solvent (e.g., MeOH) ​ The composition containing the salt solution is exchanged with a second solvent ([[]] e.g., isopropyl alcohol (IPA)) in which the solubility of the lysinilazole sodium salt is lower.

[0158] For example, when crude lysinilazole and sodium methoxide are dissolved in methanol, any trapped impurities E and F are released into the solution. Solvent exchange to IPA gradually precipitates the lysinilazole sodium salt from the solution while retaining the impurities in the mother liquor.

[0159] When the above process was applied to the composition produced according to Example 12 (below [[[]] ) containing a mixture of lysinilazole hydrate form A and impurities E and F, analysis of the recovered solid revealed that the solvent exchange method reduced the levels of impurities E and F by 46% and 59% respectively.

[0160] The sodium salt is also very soluble in DMSO and insoluble in MTBE. Therefore, a crystallization [[[]] technique can be applied, whereby the sodium salt is dissolved in a suitable solvent (e.g., methanol or [[[]] DMSO), and then a solvent in which the salt is less soluble (e.g., MTBE) is added to induce [[[]] crystallization.

[0161] Next, the purified lysinilazole salt is dissolved and anhydrous lysinilazole is precipitated (e.g., [[[]] by adding acetic acid as described in Section 5.5.2 above), and purified anhydrous crystalline [[[]] form D of lysinilazole, characterized by an XRPD pattern substantially consistent with Figure 3, can be obtained.

[0162] 5.5.5 Solvent Exchange with Lysinilazole Lithium Salt Differences in the solubility of lithium riginirazole in various solvents can also be utilized to remove the trapped impurities E and F.

[0163] Accordingly, the present invention contemplates the use of a lithium riginirazole solvent exchange step to reduce the levels of impurities E and / or F. In this step, a composition comprising a solution of lithium riginirazole mixed with impurities E and F in a first solvent is exchanged with a second solvent in which the solubility of lithium riginirazole is lower.

[0164] Lithium riginirazole can be prepared from crude riginirazole form D and LiOH in THF / DMSO at 20 °C using a stoichiometry of 1:2 riginirazole:base. The diffraction pattern is shown in Figure 19 and indicates a crystalline substance. The rise in the baseline of the diffraction pattern may indicate some amorphous content and / or may contain a DMSO solvate.

[0165] 5.5.6 Carbon treatment Impurities E and F can be removed from the crude riginirazole composition by carbon treatment. Carbon treatment is preferably applied to a solution of the crude riginirazole mixture and may include contact of such a solution with activated carbon. Suitable solutions include alkali metal riginirazole salt solutions, such as sodium, potassium or lithium riginirazole salt solutions.

[0166] Treatment with activated carbon preferably further includes the step of removing the activated carbon by filtration. Alternatively or in addition, carbon treatment may include recirculation of the solution through an activated carbon filter cartridge. ​​​​​​​​​​​​​​

[0167] In a preferred embodiment, before the carbon treatment, an alkali metal salt solution (for example, in methanol ) is formed. After the carbon treatment of this solution, lysinilazole may precipitate (for example , as described in Section 5.5.2 above, by adding acetic acid). Suitable alkali metal salts include sodium salts, potassium salts, and lithium salts. After dissolving crude lysinilazole and sodium methoxide in methanol, it is preferably carbon-treated and then precipitated with acetic acid.

[0168] Any suitable solution and any form of activated carbon can be used, such as stirring with Norit® SX Plus and recycling the solution through an activated carbon filter cartridge (for example, Zetacarbon R53SP™ cartridge). In the latter case, a carbon load corresponding to 0.086 wt can be used to recycle through the filter for at least 2.5 hours.

[0169] The carbon treatment cycle can be repeated while monitoring the levels of impurities E and F, and continued until the levels are reduced to the target levels.

[0170] Next, purified lysinilazole is precipitated (for example, by adding acetic acid as described in Section 5.5.2 above), and a purified lysinilazole anhydrous crystalline form D characterized by an XRPD pattern substantially identical to that of Figure 3 can be obtained.

[0171] Exemplary methods including carbon treatment According to a first example, lysinilazole, or a pharmaceutically acceptable derivative, salt, hydrate ​​​​​​​​​A method for producing a substance, a solvent, a substance, a complex, a biological equivalent, a metabolite or a prodrug, comprising: , (a) subjecting 3,3'-diaminobenzidine (DAB) to a condensation reaction to produce the intermediate by-product of the lysinylazo -ol and formula (II):

[0172] [Chemical formula] and then (b) after dissolving the lysinylazole, treating the lysinylazole solution with activated carbon to remove residual DAB and / or the intermediate of formula (II), or reduce its level to obtain lysinylazole in a purified form . A method is provided.

[0173] In some embodiments of this example, the DAB in step (a) is of the formula:

[0174] [Chemical formula] contains the contaminating aminobenzidine compound (MAB).

[0175] The contaminating MAB may be present at about 0.5% or more, and when exposed to the condensation reaction in step (a), formula (IV):

[0176] [Chemical formula] of the intermediate by-product is produced.

[0177] Therefore, the DAB in step (a) is of formula (III):

[0178] [Chemical formula] In embodiments where the condensation reaction is carried out in the presence of aminobenzidine contaminants, the condensation reaction is of formula (IV):

[0179]

Chemical formula

[0180] The treatment with activated carbon in step (b) may include treating the solution with activated carbon after forming a salt solution of lysinilazole. Suitable salts include sodium salts, potassium salts, and lithium salts. Sodium salts are preferred. There is a wide range for manipulating the exact state of the imidate-DAB condensation reaction, and all such

[0181] manipulations are within the scope of the present invention (as described in section 5.2 above). The condensation reaction can be carried out at a temperature from 10 °C to 100 °C. Generally, the reaction can be carried out at the reflux temperature of the solvent at atmospheric pressure.

[0182] The reaction can be carried out in any suitable solvent that does not interfere with the reaction. Suitable solvents include methanol.

[0183] The condensation is (a) adding sodium methoxide to 4-cyanopyridine in methanol to produce an imidate compound of formula (V), and then

[0184] (b) adding the compound of formula (V) from step (a) to a mixture of DAB and acetic acid in methanol and the step of producing and the step of adding It may also contain.

[0185]

Chemical formula

[0186] In step (a), by using different combinations of alkoxides / alcohols, other imidates can be produced and used in the condensation reaction. For example, sodium ethoxide / ethanol can be used instead of sodium methoxide / methanol, and on the other hand, other cations (preferably alkali metals) can replace sodium. It can be done.

[0187] In step (b), other acids such as TFA can be used instead of acetic acid.

[0188] The inventors have surprisingly found that, despite the use of highly toxic 3,3'-diaminobenzidine (DAB) and potentially toxic intermediate by-products of formula (II), the large-scale GMP synthesis of 2,2'-di(pyridin-4-yl)-1H,1'H-5,5' -bibenzimidazole suitable for use in pharmaceutical formulations can be achieved by reducing the aforementioned toxic compounds to acceptable levels using activated carbon.

[0189] Other exemplary embodiments are as defined in the following numbered paragraphs.

[0190] 1. A method for producing riginirazole, or a pharmaceutically acceptable derivative, salt, hydrate, solvate, complex, biological equivalent, metabolite or prodrug thereof, comprising: (a) subjecting 3,3'-diaminobenzidine (DAB) to a condensation reaction to obtain the riginirazole and formula (II):​​

[0191]

Chem.

[0192] 2. The DAB in step (a) is present in the condensation reaction together with the aminobenzidine contaminants of formula (III):

[0193]

Chem.

[0194]

Chem.

[0195] 3. The method according to paragraph 1 or 2, wherein the treatment with activated carbon in step (b) further comprises the step of treating the solution after forming a solution of a salt of ridinilazole, for example, a sodium, potassium or lithium salt, with activated carbon

[0196] 4. The method according to paragraph 3, wherein the salt solution of ridinilazole is a sodium salt dissolved in methanol

[0197] ​​​​​​​​5. The sodium salt of ridinilazole is formed by treatment with sodium methoxide using the method described in paragraph 4.

[0198] 6. The method according to any one of the preceding paragraphs, wherein the treatment with activated carbon in step (b) further comprises a step of removing the activated carbon by filtration .

[0199] 7. The method according to any one of paragraphs 1 to 5, wherein the treatment with activated carbon in step (b) comprises recirculation of the solution through an activated carbon filter cartridge .

[0200] 8. The method according to any one of the preceding paragraphs, wherein the treatment with activated carbon in step (b) further comprises a step of acidifying to obtain ridinilazole in a purified form .

[0201] 9. The treatment with activated carbon in step (b) (i) forming the sodium salt of ridinilazole in methanol by treatment, for example, with sodium methoxide , and (ii) treating the solution obtained in step (i) with activated carbon, and (iii) acidifying to obtain ridinilazole in a purified form and comprises the method according to any one of the preceding paragraphs.

[0202] 10. The method according to any one of the preceding paragraphs, wherein the treatment with activated carbon in step (b) reduces the level of the intermediate by - product of formula (II) to less than 100 ppm .

[0203] 11. The method according to any one of paragraphs 2 to 10, wherein the treatment with activated carbon in step (b) reduces the level of the compound of formula (IV) to less than 50 ppm .

[0204] 12. In step (a), the condensation involves reacting DAB with a compound of formula (V):

[0205]

Chemical formula

[0206] 13. In step (a), the condensation involves (a) adding sodium methoxide to 4-cyanopyridine to produce a compound of formula (V) and then (b) adding the compound of formula (V) from step (a) to DAB The method according to paragraph 12, comprising.

[0207] 14. In step (a), the condensation involves (a) adding sodium methoxide to 4-cyanopyridine in methanol to produce a compound of formula (V) and then (b) adding the compound of formula (V) from step (a) to a mixture of DAB and acetic acid in methanol or (c) adding a mixture of DAB and acetic acid in methanol to the compound of formula (V) from step (a) The method according to paragraph 13, comprising. Including.

[0208] 15. (a) Mixing purified lysinilazole in methanol / water to obtain a solid and (b) separating the solid (c) drying the solid The method according to any one of the preceding paragraphs, further comprising the step of isolating lysinilazole by. One of.

[0209] 16. In step (a), the ratio of methanol: water is from 1:2 to 1:4, for example about 1:3, the method according to paragraph 15.

[0210] 17. In step (a), the purified liginirazole is stirred in methanol / water the method according to paragraph 15 or 16.

[0211] 18. In step (a), the purified liginirazole is mixed with 10 - 40, for example about 20 volumes of methanol / water, the method according to any one of paragraphs 15 to 17 。

[0212] 19. In step (b), the solid is separated by filtration, according to any one of paragraphs 15 to 18 the method described.

[0213] 20. In step (c), the solid is dried in a filter dryer and optionally the level of water and / or methanol is monitored, according to any one of paragraphs 15 to 19 the method described.

[0214] 21. The condensation is carried out at a temperature of 30 - 80 °C, according to any one of the preceding paragraphs the method described.

[0215] 22. The condensation is carried out at a temperature of about 60 °C, the method according to paragraph 21.

[0216] 23. Further comprising the step of forming a pharmaceutically acceptable derivative, salt, hydrate, solvate, complex of the liginirazole, biological equivalent, metabolite or prodrug, according to any one of the preceding paragraphs the method described.

[0217] 24. For example, using DMSO, the step of forming a solvate of the liginirazole The method according to paragraph 23, further comprising.

[0218] 25. For manufacturing a pharmaceutical composition, the method according to any one of the preceding paragraphs, further comprising formulating purified lizinirazole in a pharmaceutically acceptable excipient. The method according to any one of the preceding paragraphs, further comprising formulating purified lizinirazole in a pharmaceutically acceptable excipient. .

[0219] 26. The method according to paragraph 25, further comprising configuring the pharmaceutical preparation in a pharmaceutical kit, a pharmaceutical pack or a patient pack. The method according to paragraph 25, further comprising configuring the pharmaceutical preparation in a pharmaceutical kit, a pharmaceutical pack or a patient pack.

[0220] 5.5.7 Polymorphic transformation In a preferred embodiment, lizinirazole is present in the crude lizinirazole composition as the anhydrous crystalline form D characterized by an XRPD pattern substantially in agreement with Figure 3. In a preferred embodiment, lizinirazole is present in the crude lizinirazole composition as the anhydrous crystalline form D characterized by an XRPD pattern substantially in agreement with Figure 3. And the removal step (b) includes a polymorphic transformation from form D to form A.

[0221] In such an embodiment, the polymorphic transformation may include slurrying the crude lizinirazole composition in an aqueous solvent and seeding the slurry with crystals of lizinirazole form A at a water activity (A ) and temperature favorable for the crystallization of lizinirazole form A. w ) and temperature favorable for the crystallization of lizinirazole form A. And seeding the slurry with crystals of lizinirazole form A.

[0222] The seeds of form A used in the seeding step can take any physical form. Thus, they are (a) micronized, (b) in the form of a dry powder, or (c) in the form of a slurry. in the form of a slurry.

[0223] A w is preferably 0.4 or more, and / or the temperature is 2 to 60 °C, more preferably A w is 0.4 to 0.5, the temperature exceeds 2 °C and is less than 30 °C, and even more preferably A wis 0.4 to 0.5, and the temperature is room temperature.

[0224] Any suitable aqueous solvent can be used. In a preferred embodiment, the solvent is MeOH / H 2O.

[0225] For example, characterized by an XRPD pattern substantially consistent with FIG. 3, and containing a mixture of impurities E and F together with the anhydrous crystalline form D of lizinilazole, the crude lizinilazole product prepared along reaction scheme 1 (above ) is slurried in an aqueous solvent the crude lizinilazole composition, and then the water activity (A ) and temperature favorable for crystallization of lizinilazole form A are used to seed the slurry with crystals of lizinilazole form A (the crystals may be micronized, added as a dry powder or added in the form of a slurry).) and is thereby converted to lizinilazole form A. w ) and temperature, crystals of lizinilazole form A (the crystals may be micronized, added as a dry powder or added in the form of a slurry).) and is thereby converted to lizinilazole form A. ) is thereby converted to lizinilazole form A.

[0226] It has been found that the above polymorph conversion procedure from the exemplary form D to form A reduces the level of impurity F by about 60% .

[0227] Exemplary lizinilazole D to A polymorph conversion process The conversion can be carried out as follows: 1) Add form D. 2) Add MeOH. 3) Heat to 60 °C. Stir at 300 rpm. 4) Hold for 15 minutes. 5) Add water over 30 minutes, a w about 0.47 6) Cool to 40 °C over 2 hours. 7) Seed with 2 wt% of form A (or seed the slurry with 2 wt% of form A prepared in MeOH / H2O (80 vol / 20 vol) and slurried for 2.5 hours before addition). ) 8) Wait for 1 hour. The slurry is thick and has limited mobility. 9) Cool to 20 °C over 2 hours. 10) Heat to 40 °C over 4 hours. 11) Cool to 20 °C over 10 hours. 12) Wait for 2.5 hours. The slurry is thick and mobile. 13) Vacuum filtration. Filtration time: 15 seconds. 14) Wash the reactor three times with 1 volume of MeOH / H2O (80 vol / 20 vol), 3 ml each time. Wash the wet cake with 1 volume of MeOH / H2O (80 vol / 20 vol), 3 ml. The wet cake is 3 ml.

[0228] In the event that polymorphic transformation is not complete, or when Form N is produced (presumably due to local variations in water activity), a reslurrying process can be carried out. Therefore, prior to the polymorphic transformation process, it is preferable to have a high-temperature methanol reslurrying step that converts any existing forms (including Form N if present) to Form D.

[0229] A preferred methanol reslurrying process is shown below: · Add methanol (7.4 vol) to the reactor. · Add crude liginilazole as a wet cake to the reactor. · Heat the slurry at 55 - 60 °C for at least 3 hours. · Cool to 20 - 25 °C and stir for at least 3 hours. · Filter the slurry. · Wash the cake with methanol (2.8 vol) and dry under vacuum. · Vacuum dry at 40 °C for at least 24 hours.

[0230] 5.6 Crystal Forms of Liginilazole As described above, the inventors have identified three distinct crystal forms that are particularly useful in the above process (multiple​​​ The discovery of the crystal form of lysinylazole (form A) and the efficient large-scale synthesis of lysinylazole for medical use is applied.

[0231] (11.02 ± 0.2)°, (16.53 ± 0.2)°, and (13.0 ± 0.2) ° characterized by peaks in the powder X-ray diffraction pattern at 2 theta angles, the crystalline form (form A) of lysinylazole tetrahydrate is described herein.

[0232] (10.82 ± 0.2)°, (13.35 ± 0.2)°, and (19.15 ± 0.2) ° characterized by peaks in the powder X-ray diffraction pattern at 2 theta angles, and optionally (10.82 ± 0.2)°, (1 3.35 ± 0.2)°, (19.15 ± 0.2)°, (8.15 ± 0.2)°, and (2 1.74 ± 0.2)° characterized by peaks in the powder X-ray diffraction pattern at 2 theta angles, the crystalline form (form N) of lysinylazole tetrahydrate is also described herein.

[0233] (12.7 ± 0.2)°, (23.18 ± 0.2)°, and (27.82 ± 0.2)° characterized by peaks in the powder X-ray diffraction pattern at 2 theta angles, and optionally (12.7 ± 0.2)°, (23.18 ± 0.2)°, (27.82 ± 0.2)°, (19.5 ± 0.2)°, and (22.22 ± 0.2)° characterized by peaks in the powder X-ray diffraction pattern at 2 theta angles, the crystalline form (form D) of lysinylazole anhydride is also described herein.

[0234] Other embodiments and aspects of this aspect of the invention are defined as follows in the numbered paragraphs below: as follows:

[0235] 1. (11.02 ± 0.2)°, (16.53 ± 0.2)°, and (13.0 ± 0.​​​ The crystalline form (Form A) of lysinilazole tetrahydrate, characterized by a powder X-ray diffraction pattern comprising peaks characteristic of a 2-theta angle of 2)°.

[0236] 2. The crystalline Form A according to paragraph 1, characterized by an XRPD pattern substantially identical to FIG. 1.

[0237] 3. The crystalline Form A according to paragraph 1 or 2, which is substantially pure.

[0238] 4. A composition comprising at least 80 wt%, 90 wt%, 95 wt% or 99 wt% of the crystalline Form A according to any one of paragraphs 1 to 3.

[0239] 5. A powder X-ray diffraction pattern comprising peaks characteristic of 2-theta angles of (10.82 ± 0.2)°, (13.35 ± 0.2)° and (19.15 ± 0. 2)°, optionally including peaks characteristic of 2-theta angles of (10.82 ± 0.2)°, (1 3.35 ± 0.2)°, (19.15 ± 0.2)°, (8.15 ± 0.2)° and (2 1.74 ± 0.2)°, characterizing the crystalline form (Form N) of lysinilazole tetrahydrate.

[0240] 6. The crystalline Form N according to paragraph 5, characterized by an XRPD pattern substantially identical to FIG. 2.

[0241] 7. The crystalline Form N according to paragraph 5 or 6, which is substantially pure.

[0242] 8. A composition comprising at least 80 wt%, 90 wt%, 95 wt% or 99 wt% of the crystalline Form N according to any one of paragraphs 5 to 7.

[0243] 9. Characterized by peaks characteristic of two theta angles of (12.7 ± 0.2)°, (23.18 ± 0.2)° and (27.82 ± 0.2 )°, and optionally containing peaks characteristic of two theta angles of (12.7 ± 0.2)°, (23. 18 ± 0.2)°, (27.82 ± 0.2)°, (19.5 ± 0.2)° and (22. 22 ± 0.2)°, the crystalline form (Form D) of lizinilazole characterized by a powder X-ray diffraction pattern containing the same.

[0244] 10. The crystalline form D described in paragraph 9, characterized by an XRPD pattern that substantially coincides with FIG. 3 as described.

[0245] 11. The crystalline form D described in paragraph 9 or 10, which is substantially pure.

[0246] 12. A composition comprising at least 80% by weight, 9 0% by weight, 95% by weight or 99% by weight of the crystalline form D described in any one of paragraphs 9 to 11.

[0247] 13. The crystalline form or composition described in any one of the preceding paragraphs, wherein XRPD is measured by Cu-K alpha radiation having a wavelength of 0.15419 nm as described.

[0248] 14. The crystalline form or composition described in paragraph 13, wherein XRPD is measured at room temperature.

[0249] 15. A method for producing the crystalline form or composition defined in any one of paragraphs 1 to 4 comprising: (a) providing a slurry of lizinilazole Form D in an aqueous solvent; and (b) seeding the slurry with crystals of lizinilazole Form A or Form N at a water activity (A w ) and temperature favorable for the crystallization of lizinilazole Form A as described.

[0250] 16.A w is 0.4 or more, and / or the temperature is 2 to 60 °C, optionally , A w is 0.4 to 0.5, the temperature exceeds 2 °C and is less than 30 °C, for example, A w is 0.4 to 0.5, and the temperature is room temperature, the method according to paragraph 15.

[0251] 17. A method for producing a crystalline form or composition defined in any one of paragraphs 5 to 8 comprising: (a) providing a slurry of lysinilazole form D in an aqueous solvent; and (b) seeding the slurry with crystals of lysinilazole form A or form N at a water activity (A w ) and temperature favorable for the crystallization of lysinilazole form N. A method comprising the steps of:

[0252] 18.A w is 0.5 or more, and / or the temperature is 2 to 60 °C, optionally , A w exceeds 0.5, the temperature exceeds 2 °C and is less than 60 °C, for example, A w is 0.55 exceeds, and the temperature is room temperature, the method according to paragraph 17.

[0253] 19. The method according to any one of paragraphs 15 to 18, wherein the solvent is MeOH / H2O. Method.

[0254] 20. A crystalline form of lysinilazole tetrahydrate obtained or produced by the method according to any one of paragraphs 15 to 19. Crystal form.

[0255] 21. A pharmaceutical composition comprising an effective amount of the crystalline form or composition according to any one of paragraphs 1 to 14 or 20 and a pharmaceutically acceptable excipient. Composition.

[0256] 22. For use in treatment or prophylaxis, a crystalline form or composition according to any one of paragraphs 1 to 14 or 20.

[0257] 23. For use in the treatment or prophylaxis of CDI or CDAD, a crystalline form or composition according to any one of paragraphs 1 to 14 or 20, or a pharmaceutical composition according to paragraph 21.

[0258] 24. Use of a crystalline form or composition according to any one of paragraphs 1 to 14 or 20 in the manufacture of a pharmaceutical composition.

[0259] 5.7 Medical uses Clostridioides difficile - associated diseases (CDAD) define a spectrum of symptoms and are associated with Clostridioides difficile (C. difficile) infection (CDI). CDAD includes diarrhea, abdominal distension, influenza - like symptoms, fever, anorexia, abdominal pain, nausea, dehydration symptoms, and enteritis (colitis). The most severe symptom of CDAD is pseudomembranous colitis (PMC), which histologically is colitis with mucosal plaques and clinically presents with severe diarrhea, abdominal cramps, and systemic toxic symptoms. The rilidilazole polymorphs / crystalline forms and pharmaceutical compositions of the present invention are applied to the treatment of all forms of CDAD such as diarrhea, abdominal distension, influenza - like symptoms, fever, anorexia, abdominal pain, nausea, dehydration symptoms, colitis, pseudomembranous colitis, etc.

[0260] 5.8 Pharmacokinetics The pharmaceutical compositions of the present invention can be administered by oral or parenteral routes such as intravenous, intramuscular, intraperitoneal, subcutaneous, transdermal, airway (aerosol), rectal, vaginal, topical (including buccal and sublingual) administration. ​​

[0261] The amount of pharmaceutical composition administered varies widely depending on the specific dosage unit used, the duration of treatment, the age and sex of the patient being treated, and the nature and extent of the disorder being treated.

[0262] Generally, the effective amount of the pharmaceutical composition administered generally ranges from about 0.01 mg / kg to 10000 mg / kg per day. The unit dosage may contain 0.05 to 500 mg of risedronate and can be taken once or multiple times per day.

[0263] The preferred route of administration is oral administration. Generally, an appropriate dosage ranges from 0.01 to 500 mg per kilogram of the recipient's body weight per day.

[0264] The desired dosage is preferably presented as a single dose for daily administration. However, two, three, four, five or six or more sub-doses administered at appropriate intervals throughout the day may be used. These sub-doses can be, for example, from 0.001 to 100 mg per unit dosage form, preferably from 0.01 to 10 mg, and most preferably from 0.5 to 1.0 mg of the active ingredient and administered in unit dosage forms containing the active ingredient.

[0265] When determining the effective amount or dosage, several factors including, but not limited to, the potency and duration of action of the compound used, the nature and severity of the disease being treated, and the sex, age, weight, general health status, individual response, and other relevant circumstances of the patient being treated are considered by the attending physician. One of ordinary skill in the art will appreciate that the dosage is determined by Goodman and Gilman's "The Pharmacological Basis of Therapeutics," 11th Edition, McGraw-Hill, New York, 2006, and Gilman's "The Pharmacological Basis of Therapeutics," 11th Edition, McGraw-Hill, New York, 2006, & Goldman's The Pharmacological Basis of Therapeutics, Ninth Edition (1996), App endix II, pp. 1707-1711. It will be understood that it can also be determined by the guidelines from

[0266] The effectiveness of a specific dosage of the pharmaceutical composition of the present invention can be determined by monitoring the effect of a given dosage on the progression of CDI and / or CDAD.

[0267] 5.9 Formulations The pharmaceutical composition may contain a stabilizer, an antioxidant, a coloring agent and a diluent. Pharmaceutically acceptable carriers and additives are selected such that side effects from the pharmaceutical compound are minimized and the performance of the compound is not impaired to the extent that the treatment becomes ineffective.

[0268] Oral (intragastric) is a typical route of administration. Pharmaceutically acceptable carriers may be solid dosage forms including tablets, capsules, pills and granules, which can be prepared with coating agents and shell agents such as enteric coating agents and others well known in the art. Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups and elixirs.

[0269] When administered, the pharmaceutical composition may be at body temperature or near body temperature.

[0270] Compositions for oral use can be prepared according to any method known in the art for the manufacture of pharmaceutical compositions, and such compositions provide pharmaceutically smooth and palatable formulations, and thus are selected from one or more of the group consisting of sweetening agents, flavoring agents, coloring agents and preservatives. ​​​​​​​​​It can contain multiple drugs. The tablets contain an active ingredient mixed with excipients that are suitable for tablet manufacturing, non-toxic, and pharmaceutically acceptable. These excipients can be, for example, inert diluents such as calcium carbonate, sodium carbonate, lactose, calcium phosphate or sodium phosphate, granulating agents and disintegrants, such as corn starch, or alginic acid, binders, such as starch, gelatin or gum arabic, and lubricants, such as magnesium stearate, stearic acid, or talc. The tablets may or may not be coated, and may be coated using known techniques, for example, to delay disintegration and absorption in the digestive tract, thereby providing a sustained action over a long period. For example, time-delay materials such as glyceryl monostearate or glyceryl distearate can be used.

[0271] Formulations for oral use may be present as hard gelatin capsules in which the active ingredient is mixed with an inert solid diluent, such as calcium carbonate, calcium phosphate or kaolin, or the active ingredient may be present by itself, or the active ingredient may be present as a soft gelatin capsule mixed with water or an oily medium, such as peanut oil, liquid paraffin or olive oil.

[0272] An aqueous suspension containing the active substance in a mixture with excipients suitable for the production of an aqueous suspension can be produced. Such excipients include suspending agents, such as sodium carboxymethylcellulose, methyl cellulose, hydroxypropylmethylcellulose, sodium alginate, polyvinyl pyrrolidone, tragacanth gum and acacia gum, and the dispersing or wetting agent is natural ​Naturally occurring phosphatides such as lecithin, or condensation products of alkylene oxides and fatty acids, such as polyoxyethylene stearate, or condensation products of ethylene oxide and long-chain fatty alcohols, such as heptadecaethyleneoxy cetyl alcohol, or condensation products of ethylene oxide with partial esters derived from fatty acids and hexitols such as polyoxyethylene sorbitol monooleate, or partial esters derived from fatty acids and hexitol anhydrides such as polyoxyethylene sorbitan monooleate may be used. The aqueous suspension may contain one or more preservatives such as ethyl or n-propyl p-hydroxybenzoate, one or more colorants, one or more flavoring agents, or one or more sweetening agents such as sucrose or saccharin. The oily suspension can be formulated by suspending the active ingredient in an omega-3 fatty acid, a vegetable oil such as peanut oil, olive oil, sesame oil or coconut oil, or a mineral oil such as liquid paraffin. The oily suspension may contain a thickening agent such as beeswax, hard paraffin or cetyl alcohol. Oral formulations with good taste can be provided by adding the above-mentioned sweetening agents and flavoring agents. These compositions can be preserved by adding an antioxidant such as ascorbic acid. Powders and granules with suitable dispersibility for preparing an aqueous suspension by adding water are provided by mixing a dispersant or wetting agent, a suspending agent, and one or more preservatives with the active ingredient.

[0273]

[0274]

[0275]

[0276] ​​​​​​​​​​​​ Examples of the cutting dispersant or wetting agent and the suspending agent are as described above. Additional excipients, such as sweeteners, flavoring agents, and coloring agents, may be present.

[0277] Syrups and elixirs containing lysinirazole can be formulated with sweeteners such as glycerol, sorbitol, or sucrose. Such formulations can also contain lubricants, preservatives, flavoring agents, and coloring agents.

[0278] The composition of the present invention can optionally be supplemented with additional agents such as, for example, viscosity enhancers, preservatives, surfactants, and penetration enhancers. Examples of viscosity enhancers include, for example, polyvinyl alcohol, polyvinylpyrrolidone, methylcellulose, hydroxypropylmethylcellulose, hydroxyethylcellulose, carboxymethylcellulose, hydroxypropylcellulose, or other agents known to those skilled in the art. Such agents are usually used at levels from about 0.01% to about 2% by weight of the pharmaceutical composition.

[0279] Preservatives are optionally used to prevent the growth of microorganisms before or during use. Suitable preservatives include polyquaternium-1, benzalkonium chloride, thimerosal, chlorobutanol, methylparaben, propylparaben, phenylethyl alcohol, disodium edetate, sorbic acid, or other agents known to those skilled in the art. Usually, such preservatives are used at levels from about 0.001% to about 1.0% by weight of the pharmaceutical composition. level.

[0280] The solubility of the components of the composition of the present invention can be adjusted by surfactants or other suitable co-solvents in the composition. can be enhanced. Such co-solvents include polysorbate 20, 60, and 80, poly oxyethylene / polyoxypropylene surfactants (e.g., pluronic F-68, F-84, and P-103), cyclodextrin, or other agents known to those skilled in the art. Typically, such co-solvents are used at levels from about 0.01% to about 2% by weight of the pharmaceutical composition.

[0281] Pharmaceutically acceptable excipients and carriers include all of the foregoing and the like. The foregoing considerations regarding effective formulation and administration procedures are well known in the art and are described in standard textbooks. See, for example, Remington: The Science and Practice of Pharmacy, 20th Edition (Lippincott, Williams and Wilkins), 2000; Lieberman et al., ed., Pharmaceutical Dosage Forms, Marcel Decker, New York, N.Y. (1980) and Kib be et al., ed., Handbook of Pharmaceutical Excipients (3rd Edition), American Ph armaceutical Association, Washington (1999).

[0282] Thus, in embodiments where the compounds of the invention are formulated with pharmaceutically acceptable excipients, for example, any suitable excipients including inert diluents, disintegrants, binders, lubricants, sweetening agents, flavoring agents, coloring agents, and preservatives can be used. Suitable inert diluents include ​​, sodium carbonate and calcium carbonate, sodium phosphate and calcium phosphate, and lactose are included, while corn starch and alginic acid are suitable disintegrants. Examples of binders include starch and gelatin, and in the presence of a lubricant it is generally magnesium stearate, stearic acid or talc. The pharmaceutical composition can take any suitable form, for example, tablets, elixirs, capsules agents, solutions, suspensions, powders, granules, nail lacquers, varnishes and veneers, skin patches and aerosols.

[0283] The pharmaceutical composition may be in the form of a kit of parts, which kit may include instructions for use together with the composition of the present invention, and / or may contain a plurality of different components in unit dosage forms .

[0284] For oral administration, the pharmaceutical composition of the present invention can be formulated into solid or liquid preparations such as capsules, pills, tablets, troches, lozenges, solvents, powders, granules, solutions, suspensions, dispersions, emulsions (these solutions, suspensions , dispersions or emulsions may be aqueous or non-aqueous). Solid unit dosage forms can be, for example, ordinary hard shell or soft shell gelatin type capsules containing surfactants, lubricants, and inert fillers such as lactose, sucrose, calcium phosphate, and corn starch. Tablets for oral use may contain pharmaceutically acceptable excipients such as inert diluents, disintegrants, binders, lubricants, sweeteners, flavoring agents, coloring agents and preservatives. Suitable inert diluents include sodium carbonate and calcium carbonate, sodium phosphate and calcium phosphate, and lactose as well. On the other hand, corn starch and alginic acid are suitable disintegrants. The binders include starch and gelatin. When a lubricant is present, it is generally magnesium stearate, stearic acid or talc. If necessary, the tablets can be coated with materials such as glyceryl monostearate or glyceryl distearate to delay absorption in the gastrointestinal tract. Capsules for oral use include hard gelatin capsules in which the compounds of the present invention are mixed with a solid diluent, and soft gelatin capsules in which the active ingredient is mixed with an oil such as water

[0285] or peanut oil, liquid paraffin or olive oil. Suitable aqueous vehicles include Ringer's solution and isotonic sodium chloride. The aqueous suspensions according to the present invention may contain suspending agents such as cellulose derivatives, sodium alginate, polyvinylpyrrolidone and tragacanth gum, and wetting agents such as

[0286] lecithin. Preservatives suitable for aqueous suspensions include ethyl and

[0287] n-propyl p-hydroxybenzoate. The compounds of the present invention may be presented as liposome formulations. The pharmaceutical compositions of the present invention include conventional tablet bases such as lactose, sucrose, and corn starch, A lubricant for preventing adhesion to the surface of a die or punch, and a tablet can be formed by combining a dye, a colorant, and a flavoring agent for enhancing the aesthetics of the tablet and enabling it to be accepted by patients. It can be achieved.

[0288] Excipients suitable for use in oral liquid preparations include water and diluents of alcohols such as ethanol, benzyl alcohol, and polyethylene alcohol, with or without the addition of pharmaceutically acceptable surfactants, suspending agents, or emulsifying agents. Examples include those with or without the addition of such agents.

Examples

[0289] 7. Illustrations Next, the present invention will be described with reference to specific examples. These are merely illustrative and are for the purpose of illustration. They are not intended to limit the claimed exclusive scope or the described invention in any way.

[0290] Method Water activity (A w ) The water activity coefficient and water activity were calculated using the UNIFAC Activity Coefficient Calculator (Choy, B.; Reible, D. (1996). UNIFAC Activity Coefficient Calculator (Version 3.0, 1996) [Software]. University of Sydney, Australia and Louisiana State University, USA).

[0291] Powder X-ray diffraction (XRPD) XRPD analysis was performed using a Panalytical Xpert Pro diffractometer equipped with a Cu X-ray tube and a Pixcel detector system. The isothermal sample was analyzed in transmission mode and held between low-density polyethylene films. The XRPD program was used with 2θ ranging from 3 to 40°, a step size of 0.013°, a count time of 99 seconds, and a runtime of approximately 22 minutes. The XRPD patterns were classified using HighScore Plus 2.2c software. Carbon (Norit®) treatment: Crude riginirazole was dissolved in methanol and 30% sodium methoxide, and the resulting solution was treated with Norit® SX Plus (0 - 0.5 wt%) and the mixture was stirred. Next, Norit® was removed by filtration with a filter aid. Then, water was added to the filtrate, followed by acetic acid to precipitate the purified riginirazole. [Example 1] Production of riginirazole Form A Reaction: 4-Cyano-pyridine (0.85 kg) was added to a reaction flask, and MeOH (5.4 kg) and NAM-30 (NaOMe as a 30 wt% solution in MeOH; 0.5 eq; 0.15 kg) were charged. The resulting mixture was heated at 60 °C for 10 minutes and then cooled. This solution was added to a mixture of 3,3'-diaminobenzidine (DAB) (0.35 kg) and acetic acid (0.25 kg) in MeOH (1 l) at 60 °C for 1 hour. Next, the mixture was heated for 2 hours. The reaction mixture was cooled to ambient temperature overnight. The crystalline mass was filtered, washed with MeOH (1.4 L), and suction dried on the filter.

[0292]

[0293]

[0294] ​​​​​​​​​​​​​​​ Purification: Norit treatment was performed 4 times.

[0295] Polymorph formation: By reslurrying in 20 volumes of 1:3 water for injection:MeOH, the desired polymorph was obtained and drying was carried out for 6 days in a vacuum drying oven at ambient temperature and nitrogen purge. was carried out.

[0296] XRPD analysis showed that this method yielded the hydrated form A of lizinilazole (see Figure 1). The reflections are shown in the following table. See). The reflections are shown in the following table.

[0297] [Table 6-1]

[0298] [Table 6-2]

[0299] [Example 2] Production of lizinilazole form N The material of pattern N was separated from the crystallization experiment performed with methyl acetate / water (15 volumes, 95.3% volume: 4.7% volume). Lizinilazole (5.0 g) was heated in methyl acetate to 5 0 °C. Water was added and the mixture was held at 50 °C for 1 hour and then cooled to ambient temperature at 0.2 °C / min. 0 °C. Water was added and the mixture was held at 50 °C for 1 hour and then cooled to ambient temperature at 0.2 °C / min. temperature.

[0300] XRPD analysis showed that this method yielded the hydrated form N of lizinilazole (see Figure 2). The reflections are shown in the following table. See). The reflections are shown in the following table.

[0301] [Table 7]

[0302] [Example 3] Production of lizinilazole form D Reaction: 4-Cyano-pyridine (0.85 kg) was added to the reaction flask, along with MeOH (5 .4 kg) and NaOMe as a 30 wt% solution in MeOH; 0.5 equivalent; 0.1 5 kg (NAM-30 was charged. The resulting mixture was heated at 60 °C for 10 minutes and then cooled . This solution was added to a mixture of DAB (0.35 kg) and acetic acid (0.25 k g) in methanol (1 l) at 60 °C over 1 hour. The mixture was then heated for 2 hours. The re action mixture was cooled to ambient temperature overnight. The crystalline mass was filtered, washed with MeOH (1.4 L) , and dried by suction on the filter.

[0303] Purification: Norit treatment was performed 4 times.

[0304] XRPD analysis showed that this method yielded anhydrous lysinilazole Form D (see Figure 3 ). The reflections are shown in the following table.

[0305]

Table 8

[0306] [Example 4] Conversion of Lysinilazole Form D to Form A Lysinilazole Form D is prepared as described in Example 3. Lysinilazole Form A is prepared as described in Example 1. The seed crystals were prepared by hand-grinding and sieving . The conversion was carried out as follows: 1) Add Form D. 2) Add MeOH. 3) Heat to 60 °C. Stir at 300 rpm. 4) Hold for 15 minutes. 5) Add water over 30 minutes, aw is about 0.47 6) Cool to 40 °C over 2 hours. 7) Sow 2 wt% of Form A (or prepare in MeOH / H2O (80 vol / 20 vol) and seed a slurry prepared and slurried for 2.5 hours before addition with 2 wt% of Form A )

[0307] 8) Wait for 1 hour. The slurry is thick and has limited mobility. 9) Cool to 20 °C over 2 hours. 10) Heat to 40 °C over 4 hours. 11) Cool to 20 °C over 10 hours. 12) Wait for 2.5 hours. The slurry is thick and mobile. 13) Vacuum filtration. The filtration time is 15 seconds. 14) Wash the reactor three times with 1 volume of MeOH / H2O (80 vol / 20 vol), 3 ml each Wash the wet cake with 1 volume of MeOH / H2O (80 vol / 20 vol), 3 ml

[0308] [Example 5] Conversion of Lysinilazole Form D to Form N Prepare lysinilazole Form D as described in Example 3. Prepare lysinilazole Form A as described in Example 1. The seed crystals were prepared by hand grinding and sieving. After about 20 minutes, the microscopic images showed mostly smaller aggregates (about 20 μm), but some larger aggregates still remained (about 80 μm). XRPD analysis showed that the material was still composed of Form A.

[0309] The conversion was carried out on a 3 g scale as shown in the following table.

[0310]

Table 9

[0311] ​ This slurry was relatively thin compared to that formed in Example 4 and maintained mobility throughout the entire period. No discoloration (indicating the presence of Form D which is brown) was observed.

[0312] The above data indicate that the riginylazole form N exhibits improved rheology under seed slurry processing conditions, thereby enabling a speed - up of filtration and an improvement in de - liquidation on a larger scale to be expected.

[0313] [Example 6] Crystal structure of riginylazole tetrahydrate form N Single crystals of riginylazole form N of a quality suitable for complete structure determination were grown by vapor diffusion at 5 °C from a solution of riginylazole in dioxane / water (82 vol%: 18 vol, A is about 0.83) / DMSO using MEK as an antisolvent. The crystal structure was determined to be monoclinic w system and space group P21 / C. The crystal structure of riginylazole tetrahydrate form N was completely solved. The crystal structure is a tetrahydrate containing half a molecule of dinylazole and two independent water molecules per asymmetric unit. Figures 5 and 6 show the contents of the asymmetric unit and the hydrogen - bonding pattern of the determined crystal structure, respectively.

[0314]

[0315] [Example 7] Crystal structure of riginylazole tetrahydrate form A Single crystals of riginylazole form A were grown via liquid diffusion at room temperature of a solution of riginylazole in NMP / di oxane using chloroform as an antisolvent. For X - ray crystal analysis at the beamline 119 of the Diamond Light Source, crystals of approximately dimensions 0.380 mm × 0.01 A 5 mm × 0.010 mm acicular crystal sample was used.

[0316] The atom-numbered arrangement of rilginirazole and water molecules is shown in Fig. 7 as an ORTEP plot. The packing diagrams of the rilginirazole form A structure are shown in Figs. 8 to 10 along each crystal axis. The hydrogen bond between rilginirazole molecules cannot be described because only one hydrogen bond between N24-H24···N51 can be clearly identified. The other hydrogen bonds that occur within the structure are formed between water molecules, imidazole hydrogens, and pyridine nitrogen atoms. However, due to the large disorder of water molecules and their hydrogen atoms, the hydrogen bond network cannot be fully resolved.

[0317] [Example 8] Crystal Structure of Rilginirazole Anhydrous Form D Single crystals of rilginirazole form D were grown via vapor diffusion at room temperature of a solution of rilginirazole in ethanol using water as an antisolvent and presented for single crystal structure determination. For X-ray crystallographic analysis, a prismatic crystal sample with approximate dimensions of 0.3 mm x 0.2 mm x 0.1 mm was used.

[0318] The structure was solved by a given automatic direct method, and all uniquely measured F2 values were refined by least-squares refinement. The numbered arrangement used for the refinement is shown in Fig. 11. The atom-numbered arrangement of the rilginirazole molecule is shown in Fig. 11 as an ORTEP plot. The packing diagrams of the rilginirazole form D structure are shown in Figs. 12 to 14 along each crystal axis. The hydrogen bonds between rilginirazole molecules form a two-dimensional network along the ab plane (see Fig. 15). The hydrogen bonds are between the donating hydrogen imidazole nitrogen atom and the accepting pyridine nitrogen atom. ​​​​​​​​​​is formed between them. This network is extended in a third direction by weaker interactions between hydrogen atoms and the π electrons of aromatic carbons. is extended in a third direction by weaker interactions.

[0319] [Example 9] Comparison of the crystal structures of lysinilazole forms A, N, and D The main difference found in the three crystal structures is that the conformation of the lysinilazole molecule is syn in form A whereas it is anti in forms N and D (see Fig. 16).

[0320] Another difference lies in the arrangement of the hydrogen bonds. Form A shows hydrogen bonds between lysinilazole molecules whereas in form N, the lysinilazole molecules interact only with water molecules. In form A a larger water channel containing four independent water molecules is seen, but as in the case of form N all channels contain two independent water molecules. In form D, since no water molecules are present only hydrogen bonds are formed between lysinilazole molecules (see Fig. 17).

[0321] A large difference is also seen in the torsion angles made between the three structures between the phenyl rings . For forms N and D, the torsion angle is equal to 180°, so the lysinilazole molecules are planar (the center of symmetry between the phenyl rings), whereas for form A, the torsion angles are 43.0 and 43.3° (for two independent molecules).

[0322] An even larger difference between the two structures is that both are different tautomers of lysinilazole, where hydrogen is bonded to N11 in form N whereas hydrogen is bonded to N 8 of the imidazole ring in form D. Since these are both hydrogen bond donors in both structures, The packing between the two structures is very different.

[0323] [Example 10] Rizinirazole tablet dosage form with Form A Rizinirazole tablets (200 mg) were prepared as follows.

[0324] Wet granulation After sieving into the bowl of a high-shear granulator, a batch quantity of rizinirazole (Form A ), lactose monohydrate, microcrystalline cellulose, hydroxypropyl cellulose, and sodium croscarmellose for the intragranular phase of wet granulation are first subjected to a short preliminary mixing at 80 revolutions per minute (rpm) for about 1 minute.

[0325] While continuing the mixing, purified water is added. When 12% by weight of water is added and when 24% by weight of water is added, the wet mass is manually transferred through a 2000 μm sieve to improve the distribution of water and returned to the bowl of the granulator each time to continue granulation. When about 35% by weight of water is added, the wet granules are transferred to a fluidized bed dryer. When that occurs, the wet granules are transferred to a fluidized bed dryer.

[0326] Drying The wet granules are dried in a fluidized bed dryer at an inlet air temperature of about 60 °C until the detection limit (LO D) (+0.5% of the initial drying blend value) is reached. When drying is complete, the dried granules are transferred to an appropriate-sized blender through a Comil equipped with an 1143 μm screen. When drying is complete, the dried granules are transferred to an appropriate-sized blender through a Comil equipped with an 1143 μm screen.

[0327] Final blending When 5 (or 6) batches of dried granules are completed, they are combined with lactose monohydrate , microcrystalline cellulose, and sodium croscarmellose for the extra granular phase. Transfer the calculated batch quantity through a 1000-micrometer sieve and manually into a 20-L bin containing the dried granules. In a blender, manually transfer the 20-L bin containing the ingredients at 30 rpm for 2 minutes to execute blending.

[0328] Lubrication Transfer the calculated batch quantity of magnesium stearate through a 250-micrometer sieve manually into a 20-L bin containing the final formulation. Lubrication is carried out by tipping the 20-L bin in the blender at 30 rpm for 2 minutes.

[0329] Tableting Tablet the tablets using an oval tool. Dust removal and metal checking are carried out at in-line post-tableting.

[0330] Coating Coat the tablet cores using a pan coater with Opadry® II Brown. The target weight increase for the coated tablets is 3 - 4%.

[0331] XRPD analysis Perform XRPD analysis on the rilginirazole tablets to confirm that no morphological changes occurred after tableting. One tablet was crushed with a mortar and pestle and analyzed by transmission XRPD. It was not possible to completely separate a small amount of sample coating from the crushed sample.

[0332] The XRPD trace showed that there were minor peak shifts in the sample compared to Form A, while on the other hand, there were extra peaks at approximately 12.5° 2-theta and approximately 19 - 24° 2-theta. XRPD analysis of the rilginirazole tablets, rilginirazole Form A, and the placebo mixture confirmed that these extra peaks were due to the placebo mixture (Figure 1). 8). That is, the extra peak was present in the placebo mixture (Figure 18), and thus was due to the excipient. It was such.

[0333] [Example 11] Method for producing purified lirinizole Reaction: 4-cyano-pyridine (0.85 kg) was placed in a reaction flask, and MeOH (5. 4 kg) and NaOMe (as a 30 wt% solution in MeOH; 0.5 equivalent) (0.1 5 kg) were added. The resulting mixture was heated at 60 °C for 10 minutes and then cooled.

[0334] The resulting solution was added dropwise to a mixture of DAB (0.35 kg) and acetic acid (0.25 kg ) in MeOH (1 l) at 60 °C over 1 hour and heated for 2 hours.

[0335] The reaction mixture was cooled to ambient temperature overnight. Next, the mass was filtered and then washed with MeOH (1. 4 L) and suction dried on the filter.

[0336] Purification: The crude product, Norit® SX Plus (260 g) and MeOH (6 kg) were placed in a container, and NaOMe (as a 30 wt% solution in MeOH (600 g) ) was added. Purification can also be achieved by recirculating the sodium salt solution through an activated carbon filter cartridge (e.g., R53SP™ cartridge). The resulting solution was stirred at ambient temperature, filtered through diatomite, and then washed with MeOH (2 × 500

[0337] ml). Water (118 g; 4 equivalents), followed by acetic acid (206 g), was added to the mixture . The resulting slurry was stirred at ambient temperature for 2 days. The suspension was filtered, washed with MeOH (1.4 L ), and dried. This treatment gives formula (II): ) and dried. This treatment gives formula (II):

[0338]

Chem.

[0339] Polymorph formation: When reslurried with 20 volumes of 1:3 water:MeOH, pure lizinilazole was obtained. Dried for 6 days in a vacuum drying oven at ambient temperature and nitrogen purge to obtain a solid hydrate.

[0340] X-ray powder diffraction: X-ray powder diffraction (XRPD) studies were performed on a Bruker AXS D2PHASER in Bragg-Brentano configuration using a Cu anode at 30 kV, 10 mA. Sample stage standard rotation; monochromatization with a Κb filter (0.5% Ni). Slits: fixed divergence slit 1.0 mm (= 0.61°), primary axial soller slit 2.5°, secondary axial soller slit 2.5°. Detector: linear detector LYNXEYE with a receiving slit 5° detector aperture. Measurement conditions: scan range 5 - 45° 2 θ, sample rotation 5 rpm, 0.5 s / step, 0.010° / step, 3.0 mm detector slit. No background correction or smoothing is applied to the pattern . The contribution of Cu-Kα2 is removed using Bruker software.

[0341] XRPD analysis showed that this method yielded lizinilazole hydrate form A (see Figure 1 ).

[0342] Equivalents The foregoing description details the presently preferred embodiments of the invention. Considering these descriptions Accordingly, those skilled in the art will expect that numerous modifications and variations will occur in practice . These modifications and variations are intended to be encompassed within the scope of the claims appended hereto .

Claims

1. A composition comprising a mixture of compounds, wherein the mixture comprises lysinilazole and compounds of formula (II ) and formula (IV): 【Chemical Formula 1】 and the total amount of impurity E and impurity F in the mixture is less than 100 ppm.

2. The lysinilazole is in a crystalline form (form A) of lysinilazole tetrahydrate characterized by peaks at 2 theta angles of (11.02 ± 0.2)°, (16.53 ± 0.2)° and (13.0 ± 0.2)° in a powder X-ray diffraction pattern (XRPD ) of the composition according to claim 1.

3. The composition according to claim 2, wherein the crystalline form A is characterized by an XRPD pattern that substantially coincides with FIG. 1 .

4. The composition according to claim 2 or 3, wherein the crystalline form A of lysinilazole is substantially pure .

5. The composition according to any one of claims 1 to 4, wherein the mixture comprises at least 80% by weight, 90% by weight, 95% by weight or 99% by weight of the crystalline form A according to any one of claims 2 to 4 .

6. The composition according to any one of claims 2 to 5, wherein the XRPD is measured by Cu-K alpha radiation having a wavelength of 0.15419 nm .

7. The composition according to claim 6, wherein the XRPD is measured at room temperature .

8. The composition according to any one of claims 1 to 7, wherein the amount of impurity E present in the mixture is less than 50 ppm .

9. The composition according to any one of claims 1 to 8, wherein the amount of impurity F present in the mixture is less than 50 ppm .

10. (a) The amount of impurity E present in the mixture is less than 50 ppm, and (b) the amount of impurity F present in the mixture is less than 50 ppm, the composition according to any one of claims 1 to 9 .

11. The composition according to any one of claims 1 to 9, wherein the amount of impurity E or the amount of impurity F in the mixture exceeds 50 ppm but is less than 100 ppm .

12. A method for producing the composition according to any one of claims 1 to 11, comprising: (a) providing a crude lysinilazole composition comprising a mixture of compounds, wherein the mixture comprises lysinilazole and compounds of formula (II) and (IV): and the total amount of impurity E and impurity F in the mixture exceeds 100 ppm, step and then [Chemical Formula 2] the compounds of formula (II) and (IV) are included, step, and then ​ (b) Removing impurity E and impurity F from the mixture, and the impurities present in the mixture A purified lizinilazole composition in which the total amount of impurity E and impurity F is less than 100 ppm And a step of manufacturing A method comprising.

13. The step of determining the total amount of impurity E and impurity F in the purified lizinilazole composition of step (b), and optionally the total amount of impurity E and impurity F in the crude lizinilazole composition of step (a). A method according to claim 12, further comprising the step of determining. The total amount of impurity E and impurity F in the composition. The method described in.

14. The method according to claim 13, wherein the determining step includes HPLC-MS.

15. The crude lizinilazole composition of step (a) is provided by subjecting 3,3'-diaminobenzidine ( DAB) to a condensation reaction to produce the lizinilazole. The method according to any one of claims 12 to 14.

16. The method according to claim 15, wherein the condensation reaction includes reacting DAB with an imidate. Method.

17. The imidate is of formula (V): [Chemical Formula 3] The method according to claim 16, which is methyl isonicotinate imidate.

18. In step (a), the condensation reaction is (a) Adding sodium methoxide to 4-cyanopyridine to produce a compound of formula (V). And then (b) Reacting the compound of formula (V) in step (a) with the DAB. The method according to any one of claims 15 to 17, comprising.

19. In step (a), the condensation is (a) Adding sodium methoxide to 4-cyanopyridine in methanol to produce a compound of formula (V). And then (b) Adding the compound of formula (V) in step (a) to a mixture of DAB and acetic acid in methanol, or (c) Adding a mixture of DAB and acetic acid in methanol to the compound of formula (V) in step (a). The method according to claim 18, comprising. The method according to claim 18, comprising. The method according to claim 18, comprising.

20. The condensation reaction is carried out at a temperature of 20 to 90 ° C, for example 30 to 80 ° C, for example about 60 ° C. The method according to any one of claims 15 to 19.

21. The crude lizinilazole composition of step (a) has the following reaction scheme: 【Chemical Formula 4】 The method according to any one of claims 12 to 20, which is made as shown.

22. As shown below, when DAB reacts with only 1 equivalent of imidate, the compound of formula (II) is formed, the method according to any one of claims 16 to 21. **Claim 23** 【Chemical Formula 5】 The compound of formula (IV) is made by the reaction of methylisonicotinimidate of formula (V) with monoaminobenzidine (MAB), the method according to any one of claims 17 to 22. **Claim 24** The compound of formula (IV) is made as shown in the following reaction scheme: The method according to claim 23. **Claim 25** By the removal step (b), an amount of impurity E present in the mixture is less than 50 ppm, to obtain a purified lysinilazole composition, the method according to any one of claims 12 to 24. [Chemical Formula 6] **Claim 26** By the removal step (b), an amount of impurity F present in the mixture is less than 50 ppm, to obtain a purified lysinilazole composition, the method according to any one of claims 12 to 25. **Claim 27** By the removal step (b), (a) an amount of impurity E present in the mixture is less than 50 ppm, and (b) an amount of impurity F present in the mixture is less than 50 ppm, to obtain a purified lysinilazole composition, the method according to any one of claims 12 to 26. **Claim 28** By the removal step (b), an amount of impurity E or an amount of impurity F present in the mixture exceeds 50 ppm but is less than 100 ppm, to obtain a purified lysinilazole composition, the method according to any one of claims 12 to 26. **Claim 29** The removal step (b) includes treating the crude lysinilazole composition with an imidate solution, and optionally, the imidate solution reacts with impurity E and / or impurity F and purges it / them from the mixture, the method according to any one of claims 12 to 28. **Claim 30** The removal step (b) includes redissolving the lysinilazole after dissolving the crude lysinilazole composition, the method according to any one of claims 12 to 29. **Claim 31** The removal step (b) includes forming a dissolved metal salt of lysinilazole present in the crude lysinilazole composition and then optionally precipitating dinilazole by neutralization, the method according to claim 30. **Claim 32** ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ The metal salt is an alkali metal salt, optionally sodium, potassium and lithium salts of lysinilazole, the method according to claim 31.

33. The crude lysinilazole composition is dissolved in sodium methoxide in methanol, then lysinilazole is precipitated with acetic acid, according to any one of claims 30 to 32 of the method.

34. The removing step (b) comprises dissolving the crude lysinilazole composition in a high-boiling aprotic solvent and then recrystallizing lysinilazole, according to any one of claims 12 to 33 of the method.

35. The high-boiling aprotic solvent is DMSO, the method according to claim 34.

36. The removing step (b) further comprises slow cooling of the solution and / or temperature cycling of the method according to claim 34 or 35.

37. The removing step (b) optionally comprises solvent exchange using a lysinilazole alkali metal salt selected from sodium, potassium and lithium salts of lysinilazole, according to any one of claims 12 to 36 of the method.

38. The removing step (b) comprises carbon treatment, according to any one of claims 12 to 37 of the method.

39. The carbon treatment is applied to a crude lysinilazole mixture solution, optionally an alkali metal lysinilazole salt solution, for example a sodium, potassium or lithium lysinilazole salt solution, according to claim 38 of the method.

40. The carbon treatment comprises contacting the solution with activated carbon, according to claim 39 of the method.

41. The treatment with activated carbon further comprises removing the activated carbon by filtration, according to claim 40 of the method.

42. The carbon treatment comprises recirculation of the solution through an activated carbon filter cartridge, according to any one of claims 38 to 41 of the method. Lysinilazole is (i) present in the anhydrous crystalline form D, and the method comprises a polymorphic transformation from form D to form A or (ii) present in the crystalline form N of lysinilazole tetrahydrate, and the method comprises a polymorphic transformation from form N to form D or (iii) present in the crystalline form N of lysinilazole tetrahydrate, and the method comprises a polymorphic transformation from form N to form D and then from form D to form A, the method according to any one of claims 12 to 42.

44. The polymorphic transformation comprises slurrying the crude liginirazole composition in an aqueous solvent and then Water activity (A ｗ ), which is favorable for the crystallization of riluzole Form A, and temperature, riluzole seeding the slurry with crystals of Form A, the method according to claim 43. **Claim 45** Said A ｗ is 0.4 or more and / or said temperature is 2 to 60° C., Claim 4 The method according to 4. **Claim 46** Said A ｗ is 0.4 to 0.5, and the temperature exceeds 2°C and is less than 30°C, Claim The method according to 45. **Claim 47** Said A ｗ The method according to claim 46, wherein A is 0.4 to 0.5 and the temperature is room temperature. **Claim 48** wherein the solvent is MeOH / H ２ O, the method according to any one of claims 44 to 47 Method. **Claim 49** The seed of Form A is (a) micronized, (b) in the form of a dry powder, or ( c) in the form of a slurry, the method according to any one of claims 44 to 48. **Claim 50** The removal step (b) is (i) reprecipitating liginirazole after dissolving the crude liginirazole composition as described in any one of claims 30 to 33; (ii) recrystallizing liginirazole after dissolving the liginirazole reprecipitated in step (i) as described in any one of claims 34 to 36; (iii) subjecting the liginirazole recrystallized in step (ii) to carbon treatment to obtain anhydrous liginirazole crystal Form D characterized by an XRPD pattern substantially identical to FIG. 3; (iv) converting Form D liginirazole to Form A by polymorphic transformation as described in any one of claims 43 to 49 The method according to any one of claims 12 to 49, comprising **Claim 51** The composition according to any one of claims 1 to 11 obtainable (or produced) by the method according to any one of claims 12 to 50. **Claim 52** A pharmaceutical composition comprising an effective amount of the composition according to any one of claims 1 to 11 or 51 and a pharmaceutically acceptable excipient. **Claim 53** The composition according to any one of claims 1 to 11 and 51 or 52 for use in treatment or prevention. **Claim 54** The composition according to any one of claims 1 to 11 and 51 to 53 for use in the treatment or prevention of CDI or CDAD. **Claim 55** Use of the composition according to any one of claims 1 to 11 and 51 to 54 for the manufacture of a medicament for the treatment, therapy or prevention of CDI or CDAD. **Claim 56** A crystalline form (Form A) of liginirazole tetrahydrate characterized by a powder X-ray diffraction pattern comprising peaks characteristic of 2 theta angles of (11.02 ± 0.2)°, (16.53 ± 0.2)°, and (13.0 ± 0.2) °. ​

57. The crystalline form A according to claim 56, characterized by an XRPD pattern that substantially coincides with FIG.

1.

58. The crystalline form A according to claim 56 or 57, which is substantially pure.

59. A composition comprising at least 80 wt%, 90 wt%, 95 wt% or 99 wt% of the crystalline form A according to any one of claims 56 to 58.

60. The crystalline form A according to any one of claims 56 to 58 or the composition according to claim 59, in the form of micronized seeds.

61. In the method according to any one of claims 12 to 50, for use in the step of polymorphically converting lysinilazole form D to lysinilazole form A, for example, as described in any one of claims 43 to 49, the crystalline form A or composition according to any one of claims 56 to 60.

62. Use of the crystalline form A or composition according to any one of claims 56 to 60 in the manufacture of the composition according to any one of claims 1 to 11 and claims 51 to 54.

63. Characterized by peaks characteristic of 2 theta angles of (12.7 ± 0.2)°, (23.18 ± 0.2)° and (27.82 ± 0.2)°, optionally including peaks characteristic of 2 theta angles of (12.7 ± 0.2)°, (23.18 ± 0.2)°, (27.82 ± 0.2)°, (19.5 ± 0.2)° and (22.22 ± 0.2)°, the crystalline form (form D) of lysinilazole anhydrate characterized by a powder X-ray diffraction pattern.

64. The crystalline form D according to claim 63, characterized by an XRPD pattern that substantially coincides with FIG.

3.

65. The crystalline form D according to claim 63 or 64, which is substantially pure.

66. A composition comprising at least 80 wt%, 90 wt%, 95 wt% or 99 wt% of the crystalline form D according to any one of claims 63 to 65.

67. In the method according to any one of claims 12 to 50, for use in the step of polymorphically converting lysinilazole form D to form A, for example, as described in any one of claims 43 to 49, the crystalline form D or composition according to any one of claims 63 to 66.

68. In the manufacture of the composition according to any one of claims 1 to 11 and 51 to 54, Use of the crystalline form D or composition according to any one of claims 63 to 66.

69. In the production of the composition according to any one of claims 1 to 11 and 51 to 54, Use as an intermediate of the crystalline form D or composition according to any one of claims 63 to 66 Use.

70. (10.82 ± 0.2)°, (13.35 ± 0.2)° and (19.15 ± 0.2) ° including peaks characteristic of 2-theta angles, optionally (10.82 ± 0.2)°, (13. 35 ± 0.2)°, (19.15 ± 0.2)°, (8.15 ± 0.2)° and (21. 74 ± 0.2)° characterized by a powder X-ray diffraction pattern including peaks characteristic of 2-theta angles The crystalline form (form N) of lysinilazole tetrahydrate.

71. The crystalline form N according to claim 70, characterized by an XRPD pattern substantially coinciding with FIG. 2 The crystalline form N.

72. The crystalline form N according to claim 70 or 71, which is substantially pure.

73. A composition comprising at least 80% by weight, 90 % by weight, 95% by weight or 99% by weight of the crystalline form N according to any one of claims 70 to 72.

74. In the method according to any one of claims 12 to 50, for example, as described in claims 43 to 4 9, the crystalline form N or composition according to any one of claims 7 0 to 73 for use in the lysinilazole polymorph conversion step.

75. In the production of the composition according to any one of claims 1 to 11 and 51 to 54, Use of the crystalline form N or composition according to any one of claims 70 to 73.

76. In the production of the composition according to any one of claims 1 to 11 and 51 to 54, Use as an intermediate of the crystalline form N or composition according to any one of claims 70 to 73 Use.

77. The XRPD is measured by Cu-K alpha radiation having a wavelength of 0.15419 nm The crystalline form, composition, or use according to any one of claims 56 to 76 。

78. The crystalline form or composition according to claim 77, wherein the XRPD is measured at room temperature.

79. Alkali metal salts of lysinilazole.

80. The alkali metal salt according to claim 79, selected from the sodium, lithium and potassium salts of lysinilazole The alkali metal salt.

81. In the production of the composition according to any one of claims 1 to 11 and 51 to 54, The alkali metal salt according to claim 79 or 80 for use as an intermediate.

82. In the production of the composition according to any one of claims 1 to 11 and 51 to 54, Use of the alkali metal salt according to any one of claims 79 to 81 as an intermediate.

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