Integrin-inhibiting small molecule compound

Abstract

The purpose of the present invention is to provide a novel compound group having integrin function-inhibiting activity, a pharmaceutical composition, and a prophylactic agent or a therapeutic agent for diseases accompanied by fibrosis.　The present invention provides a compound represented by formula (I) (in the formula: A is C-OH or N; B is C-H, C-COOH, C-CH3, C-Br, or N; X is -SF5 or -Si(CH3)3; Y is -F or -OH; and * represents an asymmetric carbon atom), or a salt thereof. The present invention also provides an integrin-inhibiting agent using the same, a pharmaceutical composition, and a prophylactic agent or a therapeutic agent for diseases accompanied by fibrosis.

Description

Integrin-inhibiting small molecule compounds

[0001] The present invention relates to a low-molecular-weight organic compound or a salt thereof that has an integrin inhibitory function, and further to an integrin inhibitor, a pharmaceutical composition, and a therapeutic or preventive agent for diseases accompanied by fibrosis, which use the compound.

[0002] Fibrosis, a physiological phenomenon associated with aging, often spreads pathologically. However, the lack of therapeutic drugs presents a significant unsolved medical problem. Pathological fibrosis is well-known in individual organs, particularly the lungs and liver. These organs are known as interstitial pneumonia and liver cirrhosis, respectively. It is chronically (or rarely acutely) progressive, resulting in widespread and irreversible fibrosis. The pathological manifestation of fibrosis is the excessive deposition of extracellular matrix proteins, such as collagen, which replaces organ-specific parenchymal cells and eventually causes organ loss. This progressive process typically causes not only pain but also psychological fear for patients. Furthermore, even minor fibrosis, if accumulated, can damage organs and contribute to cardiac (myocardial fibrosis leading to heart failure) and renal (interstitial fibrosis leading to proteinuria) diseases. Given these factors, there is a great need for therapeutic agents that can halt and reverse fibrosis, which are expected to contribute not only to the cure of specific fibrotic diseases but also to extending healthy lifespan. Although the global race to develop the drug is heating up, there is still no cure. According to estimates from epidemiological surveys, it is predicted that at least 5 million people in Japan will develop liver disease alone over the next 10 years.

[0003] In addition to collagen, various matrix proteins are the primary structural molecules of fibrotic tissue. These proteins are produced by activated and / or differentiated fibroblasts and are regulated by TGFβ signaling. This pathology strongly suggests the involvement of TGFβ and the 24 integrin family of matrix receptors. TGFβ resides scattered and waiting in a latent form within the extracellular matrix deposits, becoming activated as needed and providing essential instructions to surrounding cells for the pathogenesis of fibrosis (e.g., matrix secretion, fibroblast activation, and differentiation into myofibroblasts). However, developing a therapeutic strategy that directly targets and inhibits the highly functional molecule TGFβ is difficult due to side effects and other factors. Integrins, on the other hand, are proteins present in the plasma membrane on the cell surface and can connect the cytoskeleton within the cell with the extracellular matrix outside the cell. Its structure is a heterodimer consisting of two subunits, an α chain and a β chain, associated in a 1:1 ratio. At least 18 types of α chains and at least 8 types of β chains have been reported. They are designated by the constituent α and β chains, such as α1β1 (integrin consisting of an α1 α chain and a β1 β chain). Integrins are known to mediate the transformation of TGFβ from its latent form to its active form. This "integrin activation" occurs when integrins bind to latent TGFβ, thereby causing tension to remove the sheath (LAP; latency associated protein) that surrounds the active TGFβ. LAP is the N-terminal portion of pro-TGFβ, which is encoded by the same gene as TGFβ. The binding site is the arginine-glycine-aspartic acid (Arg-Gly-Asp) sequence found in LAP, and only 8 of the 24 integrins (RGD-recognizing integrins) bind to this sequence. However, although there are some integrins that cannot be activated even after binding, five of the 24 types of αv-integrins (αvβ1, αvβ3, αvβ5, αvβ6, αvβ8) all recognize RGD and activate TGFβ, and therefore inhibitors of these integrins are attracting much attention as anti-fibrotic drugs.In particular, Pliant's αvβ1 / αvβ6 dual inhibitor PLN-74809 was reported in July 2022 to have proven effective in the initial Phase 2a clinical trial for interstitial pneumonia, attracting considerable attention (Non-Patent Document 1).

[0004] The first step in integrin TGFβ activation is binding to LAP. LAP contains an RGD sequence that is preferentially bound exclusively by αv-integrins. This binding triggers TGFβ to become free. Besides αv-integrins, three other RGD-recognizing integrins are known. One of these, α8β1, has been confirmed to activate TGFβ and also differentiate fibroblasts into myofibroblasts. Meanwhile, the expression site of α8β1 is significantly more limited than other integrins, being almost exclusively found in fibroblasts, and its expression level is increased by fibrotic stimuli. Thus, both its function and expression are strongly linked to the progression of fibrosis. These facts strongly suggest that the function of α8β1 plays a role in fibrosis formation. In fact, α8β1 blocking antibodies have been shown to suppress fibrosis in the liver (Non-Patent Document 2).

[0005] Pliant Therapeutics Announces Positive Safety and Efficacy Data from the Phase 2a INTEGRIS-IPF Clinical Trial of PLN-74809 in Patients with Idiopathic Pulmonary Fibrosis [online], July 10, 2022, Pliant Therapeutics Press Release, Internet<URL:https: / / ir.pliantrx.com / node / 7886 / pdf> Norihisa Nishimichi and 7 other authors, Journal of Pathology. January 12, 2021, Vol.253 issue 4, p. 366-373

[0006] In view of the above-mentioned conventional situation, an object of the present invention is to provide a novel group of compounds having activity of inhibiting the function of integrin, a pharmaceutical composition, and a therapeutic or preventive agent for diseases accompanied by fibrosis.

[0007] The present invention provides a compound represented by the following formula (I) or a salt thereof:

[0008] (Wherein, A is C—OH or N, and B is C—H, C—COOH, C—CH ３ , C—Br, or N, and X is —SF ５ , or -Si(CH ３ ) ３ Y is -F or -OH, and * represents an asymmetric carbon atom, or a salt thereof.

[0009] In the compound of the present invention or a salt thereof, X is -SF ５ and B is C—H, C—CH ３ In this case, it is preferable that Y is —F, and furthermore, it is preferable that A is C—OH. In addition, in the compound or salt thereof of the present invention, X is —Si(CH ３ ) ３ and B may be C—COOH or N.

[0010] The present invention also provides an integrin inhibitor comprising any of the above compounds or a pharmaceutically acceptable salt thereof, and a pharmaceutical composition comprising any of the above compounds or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.

[0011] The present invention also provides a therapeutic or preventive agent for a disease accompanied by fibrosis, comprising any one of the compounds or a pharmaceutically acceptable salt thereof as an active ingredient. In this case, the disease accompanied by fibrosis may be kidney fibrosis, pulmonary fibrosis (e.g., interstitial pneumonia), or liver fibrosis (e.g., cirrhosis), and the agent may be used in combination with at least one or more drugs selected from drugs classified as therapeutic or preventive agents for diseases accompanied by fibrosis. The present invention also provides a therapeutic or preventive agent for a disease accompanied by fibrosis, comprising a combination of any one of the compounds or a pharmaceutically acceptable salt thereof and the active ingredient of at least one or more drugs selected from drugs classified as therapeutic or preventive agents for diseases accompanied by fibrosis.

[0012] The present invention also provides any of the above compounds or a pharmaceutically acceptable salt thereof for use as a therapeutic or preventive agent for a disease accompanied by fibrosis. The present invention also provides use of any of the above compounds or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for treating or preventing a disease accompanied by fibrosis. The present invention also provides a method for treating or preventing a disease accompanied by fibrosis, by administering any of the above compounds or a pharmaceutically acceptable salt thereof to a patient.

[0013] The present invention includes the following aspects: [1] Formula (I)

[0014] (Wherein, A is C—OH or N, and B is C—H, C—COOH, C—CH ３ , C—Br, or N, and X is —SF ５ , or -Si(CH ３ ) ３ [2] In the formula (I), X is -SF ５ and B is C—H, C—CH ３ or C—Br. [3] The compound or salt thereof according to the above [2], wherein in formula (I), Y is —F. [4] The compound or salt thereof according to the above [3], wherein in formula (I), A is C—OH. [5] The compound or salt thereof according to formula (I), wherein X is —Si(CH ３ ) ３and B is C—COOH or N. [6] An integrin inhibitor comprising the compound according to any one of the above [1] to [5] or a pharmaceutically acceptable salt thereof. [7] A pharmaceutical composition comprising the compound according to any one of the above [1] to [5] or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier. [8] A therapeutic or preventive agent for a disease accompanied by fibrosis, comprising as an active ingredient the compound according to any one of the above [1] to [5] or a pharmaceutically acceptable salt thereof. [9] The therapeutic or preventive agent for a disease accompanied by fibrosis according to {8} above, wherein the disease accompanied by fibrosis is kidney fibrosis, lung fibrosis, or liver fibrosis.

[10] The therapeutic or preventive agent for a disease accompanied by fibrosis according to the above [8], for treating or preventing a disease accompanied by fibrosis in combination with at least one agent selected from agents classified as therapeutic or preventive agents for diseases accompanied by fibrosis.

[11] A therapeutic or preventive agent for a disease accompanied by fibrosis, comprising a combination of the compound according to any one of the above [1] to [5] or a pharmaceutically acceptable salt thereof with the active ingredient of at least one or more drugs selected from drugs classified as therapeutic or preventive agents for diseases accompanied by fibrosis.

[12] The compound according to any one of the above [1] to [5] or a pharmaceutically acceptable salt thereof for use as a therapeutic or preventive agent for a disease accompanied by fibrosis.

[13] Use of the compound according to any one of the above [1] to [5] or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for treating or preventing a disease accompanied by fibrosis.

[14] A method for treating or preventing a disease accompanied by fibrosis, comprising administering to a patient the compound according to any one of the above [1] to [5] or a pharmaceutically acceptable salt thereof.

[0015] The compound or salt thereof, or integrin inhibitor of the present invention has the activity of inhibiting integrin function, and can therefore suppress the integrin-mediated transformation of TGFβ from its latent form to its active form. Furthermore, it is expected to serve as a therapeutic agent, preventive agent, or prodrug thereof for diseases accompanied by fibrosis in which active TGFβ is involved, and can be used in the development of pharmaceuticals for fibrosis-associated diseases. The pharmaceutical composition of the present invention has the effect of treating or preventing diseases accompanied by fibrosis, using inhibition of integrin function as one of its mechanisms of action. The therapeutic agent or preventive agent for diseases accompanied by fibrosis of the present invention has the effect of alleviating symptoms of diseases accompanied by fibrosis, such as kidney fibrosis, interstitial pneumonia, or liver cirrhosis, or preventing the onset of diseases accompanied by fibrosis.

[0016] This shows the effect on TGFβ activation on the cell surface of lung fibroblasts. From the left, these are the results of measuring luciferase activity in reporter cells only, a mixed culture of reporter cells and lung fibroblasts, and when IBF17 and C8 were added. * indicates the control. * indicates P<0.05, ** indicates P<0.01, *** indicates P<0.001, and n.s. indicates no significant difference (the same applies to the following figures unless otherwise specified). Note that significant differences in this experiment were tested using Dunnett's multiple comparisons test against the control. This shows the effect on myofibroblast differentiation of fibroblasts. This shows the results of Western blotting using α-SMA and GAPDH when IBF-17 and C8 (comparison) were added. This shows the effect on myofibroblast differentiation of fibroblasts. The relative values ​​of the band intensity of α-SMA when IBF-17 and C8 (comparison) were added are shown. Significant differences were assessed by Dunnett's multiple comparisons test relative to the addition of TGFβ alone (0 nM of compound). ４ The figures show the dosing schedule for an 8-week administration experiment in a 2-week administration model. The figures also show the results of quantification of hydroxyproline in liver tissue in the same experiment. Significant differences are indicated by CCl ４The significance of the effect of carbon tetrachloride (CCl4) alone was assessed by Student's t-test. The comparison of Sirius Red stained areas of tissues in the same experiment and photographs after staining are shown. The significance of the effect was assessed by Bonferroni's Multiple Comparison test in the same experiment. The expression levels (relative intensities) of mRNA of fibrosis marker genes (Col1a1 gene and Acta2 gene) in the same experiment are shown. ４ The dosing schedule for a 3-day administration experiment in the 10-mg / kg / day (C10) administration model is shown. The mRNA expression level (relative intensity) measured by qPCR in the same experiment is shown. Significant differences are indicated by the C10 ４ (Veh) was tested by Dunnett's multiple comparisons test. Figure 1 shows the dosing schedule for an 8-week administration experiment in a NASH model. Figure 2 shows the results of quantification of hydroxyproline in liver tissue from the same experiment. Significant differences were tested by Student's t-test against CDAHFD feeding alone (0 mg / kg). (A) and (B) show the changes in body weight of mice in the same experiment and liver weight after 56 weeks. Figure 1 shows a comparison of Sirius Red-stained areas of tissues from the same experiment and photographs after staining. Significant differences were tested by Bonferroni's multiple comparisons test against CDAHFD feeding alone (0 mg / kg). The figures show the mRNA expression levels (relative intensity) of genes considered to be fibrosis markers (Acta2 gene, Col1a1 gene, and Timp-1 gene) in the same experiment. The figures show the dosing schedule of a 10-day administration experiment in a unilateral ureteral obstruction (UUO) model. The figures show the mRNA expression levels (relative intensity) by qPCR in the same experiment. Significant differences were tested by Dunnett's multiple comparisons test against UUO (Veh). The figures show the area under the blood concentration-time curve and margin when the compound of the present invention was administered. Carbon tetrachloride (CC1 ４ The figures show the dosing schedule for a 15-day administration experiment in a 15-day administration model. The figures show the results of quantification of hydroxyproline in liver tissue in the same experiment. Significant differences are indicated by CCl ４(Vehicle). The dosing schedule for a 10-day administration experiment in a bleomycin administration model is shown. The mRNA expression levels (relative intensity) of fibrosis-related genes (Acta2, Col1a1, Col3a1, Tnc) in the same experiment are shown. Significant differences were assessed by Dunnett's multiple comparisons test against Bleo (vehicle). A comparison of Masson's trichrome stained areas of lung tissue in the same experiment and photographs after staining are shown. Significant differences were assessed by Dunnett's multiple comparisons test against Bleo (vehicle).

[0017] The present invention will be described below, taking as an example exemplary embodiments, along with preferred methods and materials that can be used in carrying out the present invention, but the present invention is not limited to the embodiments described below. Unless otherwise specified, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art to which the present invention belongs. Furthermore, any materials and methods equivalent or similar to those described herein can also be used in carrying out the present invention. Furthermore, all publications and patents cited in this specification in connection with the present invention are incorporated herein by reference and constitute a part of this specification, for example, to describe methods, materials, and the like that can be used in the present invention.

[0018] In this specification, the expression "A to B" indicating a range of numerical values ​​means a range of numerical values ​​including the endpoints A and B. The same applies to "A to B." In addition, in this specification, the word "about" is used to mean that ±10% is acceptable.

[0019] 1. Compound or Salt Thereof 1-1. Structure of Compound The compound of the present invention has a chemical structure represented by the following formula (I):

[0020] In the formula, A is C—OH or N, and B is C—H, C—COOH, or C—CH ３ , C—Br, or N, and X is —SF ５, or -Si(CH ３ ) ３ and Y is —F or —OH.

[0021] In formula (I), an asymmetric carbon atom is present at the position indicated by *, and therefore the compound of the present invention exists in two optical isomers represented by the following formulae (II) and (I-II), and may be only the S-form, only the R-form, or a racemic mixture. In the following explanation, the structural formula of the S-form will be described, but the present invention is not limited to this.

[0022]

[0023] In formula (II), when A is C—OH, the chemical structure is represented by the following formula (II), and when A is N, the chemical structure is represented by the following formula (III).

[0024]

[0025] In formula (II), when B is C—H, the chemical structure is represented by the following formula (IV), and when B is C—COOH, the chemical structure is represented by the following formula (V), and when B is C—CH ３ When B is C—Br, the chemical structure is represented by the following formula (VI), when B is C—Br, the chemical structure is represented by the following formula (VII), and when B is N, the chemical structure is represented by the following formula (VIII).

[0026]

[0027] In formula (II), X is -SF ５ In this case, the chemical structure is represented by the following formula (IX), and X is -Si(CH ３ ) ３ In this case, the chemical structure is represented by the following formula (X).

[0028]

[0029] In formula (II), when Y is -F, the chemical structure is represented by the following formula (XI), and when Y is -OH, the chemical structure is represented by the following formula (XII).

[0030]

[0031] 1-2. Specific Examples of Compounds Table 1 shows specific examples of compounds having the chemical structure represented by formula (I) above.

[0032]

[0033] In IBF-17, A is C-OH, B is C-H, and X is -SF ５ and Y is -F. IBF-21 is a compound in which A is C-OH, B is C-COOH, and X is -Si(CH ３ ) ３ , and Y is —OH. IBF-18 is a compound in which A is C—OH, B is N, and X is —Si(CH ３ ) ３ and Y is —OH. IBF-20 is a compound in which A is C—OH, B is C—Br, and X is —SF ５ , and Y is —OH. IBF-13 is a compound in which A is N and B is C—CH ３ , X is -SF ５ , Y is -F.

[0034] 1-3. Salts of Compounds Salts of the compounds of the present invention may be salts with inorganic bases, organic bases, inorganic acids, organic acids, acidic or basic amino acids, etc. When the compound of the present invention represented by formula (I) has an acidic functional group, it may be a salt with an inorganic base, organic base, or basic amino acid. Furthermore, when the compound of the present invention represented by formula (I) has a basic functional group, it may be a salt with an inorganic acid, organic acid, or acidic amino acid.

[0035] Salts with inorganic bases include, but are not limited to, sodium salts, potassium salts, ammonium salts, calcium salts, magnesium salts, barium salts, aluminum salts, etc. Salts with organic bases include, but are not limited to, salts with trimethylamine, triethylamine, pyridine, picoline, 2,6-lutidine, ethanolamine, diethanolamine, triethanolamine, tromethamine [tris(hydroxymethyl)methylamine], tert-butylamine, cyclohexylamine, dicyclohexylamine, N,N-dibenzylethylamine, etc. Salts with inorganic acids include, but are not limited to, salts with hydrochloric acid, phosphoric acid, sulfuric acid, hydrobromic acid, hydroiodic acid, nitric acid, etc. Salts with organic acids include, but are not limited to, salts with acetic acid, phthalic acid, fumaric acid, oxalic acid, trifluoroacetic acid, citric acid, maleic acid, succinic acid, malic acid, acetic acid, formic acid, propionic acid, benzoic acid, methanesulfonic acid, benzenesulfonic acid, para-toluenesulfonic acid, camphorsulfonic acid, etc. Salts with acidic amino acids include, but are not limited to, salts with aspartic acid and glutamic acid, and salts with basic amino acids include salts with arginine, lysine, ornithine, etc.

[0036] The starting materials and intermediates of the compounds of the present invention may form salts. The salts of the starting materials and intermediates are preferably conventional non-toxic salts. Examples of such salts include salts with bases such as inorganic base salts (e.g., sodium salt, potassium salt, ammonium salt, calcium salt, magnesium salt, barium salt, aluminum salt, etc.) and organic base salts (e.g., trimethylamine salt, triethylamine salt, pyridine salt, picoline salt, dicyclohexylamine salt, N,N-dibenzylethylamine salt); salts with acids such as inorganic acid salts (e.g., hydrochloride, hydrobromide, hydroiodide, sulfate, nitrate, phosphate, etc.) and organic acid salts (e.g., acetate, trifluoroacetate, maleate, fumarate, citrate, tartrate, methanesulfonate, benzenesulfonate, formate, para-toluenesulfonate, etc.); and salts with amino acids (e.g., arginine, aspartic acid, glutamic acid, etc.), which can be appropriately selected by those skilled in the art.

[0037] 1-4. Method for Synthesizing Compounds The compounds can be synthesized, for example, by synthesizing the aromatic moiety on the right side of the chemical structure represented by formula (I) and the two cyclic structures (cyclic guanidine and aromatic) on the left side, and then coupling them, although this is not a limitation. The general synthesis procedure is explained in Schemes I to III below, but the following schemes and examples are merely examples of the present invention and are not intended to limit the scope of the present invention. Those skilled in the art can synthesize the compounds of the present invention by appropriately modifying the schemes based on the disclosures of this specification, existing commercially available pharmaceuticals, and known synthesis examples.

[0038]

[0039] Scheme I shows an example of a procedure for synthesizing the aromatic moiety on the right side of the chemical structure represented by formula (I). Intermediate 1 was synthesized by the condensation reaction of 2-methylpropane-2-sulfinamide with an appropriate aromatic compound containing an aldehyde group. Next, intermediate 2 was synthesized by the nucleophilic addition of methyl bromoacetate to intermediate 1 in the presence of zinc. Intermediate 2 was treated with hydrochloric acid to remove the sulfinyl group, and the hydrochloride salt of intermediate 3 was synthesized.

[0040]

[0041] Scheme II shows an example of a procedure for synthesizing the two cyclic structural moieties on the left side of the chemical structure represented by formula (I). Intermediate 4 was synthesized by reacting a suitable aromatic compound containing an amino group and a carboxylic acid group with methyl isothiocyanate. Intermediate 5 was then synthesized by reacting the thio group of intermediate 4 with iodomethane. An appropriate 1,3-diamine-2-propane derivative was reacted with intermediate 5 in hot N,N-dimethylacetamide (DMA) or N,N-dimethylformamide (DMF). The HCl salt may be obtained by lyophilization from dilute hydrochloric acid.

[0042]

[0043] Scheme III shows an example of a procedure for synthesizing a compound represented by formula (I) by coupling intermediate 3 of Scheme I with intermediate 6 of Scheme II. In Scheme III, intermediate 6 is activated for coupling using known methods and reacted with ethyl glycinate hydrochloride. The ester is hydrolyzed to an acid by treatment with a base such as NaOH in a suitable solvent (water, dioxane / water, or acetonitrile / water), followed by acidification to synthesize intermediate 7. Next, intermediate 7 is activated for coupling using known methods and reacted with intermediate 3 of Scheme I to synthesize intermediate 8. Finally, intermediate 8 is hydrolyzed to an acid by treatment with a base such as LiOH in a suitable solvent (dioxane / water or acetonitrile / water), followed by acidification to synthesize a compound represented by formula (I). In Scheme III, intermediate 6 is coupled to intermediate 3 by addition of glycine, but intermediate 3 may also be coupled to intermediate 6 by treatment with activated Boc glycine.

[0044] The compound IBF-17 can be synthesized from the following compounds using the above Schemes I to III. Here, 3-(pentafluorosulfanyl)benzaldehyde (CAS number: 401892-80-6) and 3-amino-5-hydroxybenzoic acid (CAS number: 14206-69-0) are commercially available. 2-Fluoropropane-1,3-diamine can be obtained, for example, by the synthesis method described in Example 1.

[0045]

[0046] The compound IBF-21 can be synthesized from the following compounds using the above Schemes I to III. Here, 3-amino-5-hydroxybenzoic acid and 1,3-diamino-2-propanol (CAS number: 616-29-5) are commercially available. 3-Carboxy-5-(trimethylsilyl)benzaldehyde can be synthesized according to the following Scheme IV.

[0047]

[0048]

[0049] Scheme IV shows an example of a procedure for synthesizing 3-carboxy-5-(trimethylsilyl)benzaldehyde from commercially available 1,3,5-tribromobenzene (CAS number: 626-39-1) (see Hiroaki Horiuchi et al., "Silylation Improves the Photodynamic Activity of Tetraphenylporphyrin Derivatives In Vitro and In Vivo," CHEMISTRY A Europe Journal, Supporting Information). First, n-butyllithium (n-BuLi) was added to 1,3,5-tribromobenzene in diethyl ether (EtO), and then chlorotrimethylsilane was added to synthesize 1,3-dibromo-5-(trimethylsilyl)benzene. Next, n-butyllithium (n-BuLi) was added to 1,3-dibromo-5-(trimethylsilyl)benzene in diethyl ether (EtO), followed by the addition of N,N-dimethylformamide (DMF) to synthesize 3-bromo-5-(trimethylsilyl)benzaldehyde. Furthermore, p-toluenesulfonic acid and ethylene glycol were added to 3-bromo-5-(trimethylsilyl)benzaldehyde in toluene to synthesize 2-(3-bromo-5-(trimethylsilyl)phenyl)-1,3-dioxolane. Then, 2-(3-bromo-5-(trimethylsilyl)phenyl)-1,3-dioxolane was added to activated Mg in dehydrated tetrahydrofuran (THF), followed by the introduction of carbon dioxide gas to synthesize 2-(3-carboxy-5-(trimethylsilyl)phenyl)-1,3-dioxolane. Finally, 2-(3-carboxy-5-(trimethylsilyl)phenyl)-1,3-dioxolane was dissolved in chloroform (CHCl ３ ) and HCl was added to synthesize 3-carboxy-5-(trimethylsilyl)benzaldehyde.

[0050] The compound IBF-18 can be synthesized from the following compounds using the above schemes I to III, where 3-pyridinecarboxaldehyde, 5-(trimethylsilyl) (CAS number: 144056-15-5), 3-amino-5-hydroxybenzoic acid, and 1,3-diamino-2-propanol are all commercially available and available.

[0051]

[0052] The compound IBF-20 can be synthesized from the following compounds using the above schemes I to III, where 3-bromo-5-(pentafluorosulfur)benzaldehyde (CAS number: 1240257-22-0), 3-amino-5-hydroxybenzoic acid, and 1,3-diamino-2-propanol are all commercially available and available.

[0053]

[0054] The compound IBF-13 can be synthesized from the following compounds using the above schemes I to III. Here, 3-metro-5-(pentafluorosulfur)benzaldehyde (CAS number: 1240257-01-5) and 5-aminonicotinic acid (CAS number: 24242-19-1) are commercially available, and 2-fluoropropane-1,3-diamine can be obtained, for example, by the synthesis method described in Example 1.

[0055]

[0056] Other synthetic methods include those described in, for example, JP-A-2015-524412, JP-A-2019-500355, and Neil C. Henderson et al., Nat Med. 2013 December; 19(12): 1-26, as well as principles of organic chemistry applied by those skilled in the art.

[0057] 2. Integrin Inhibitors Integrins are known to mediate the transformation of TGFβ from its latent form to its active form, and the active ingredient of the integrin inhibitor of the present invention is a compound that has the activity of inhibiting the function of integrins involved in fibrosis. Compounds represented by formula (I), such as the compounds listed in Table 1 above, or pharmaceutically acceptable salts thereof, have the activity of inhibiting the function of integrins involved in fibrosis and can be used as integrin inhibitors. The structure of the compound represented by formula (I) is as described in 1 above. Examples of integrins involved in fibrosis include αvβ1, αvβ3, αvβ5, αvβ6, αβ8, and α8β1.

[0058] In the present invention, "inhibiting integrin function" refers to inhibiting the binding of the cell membrane surface receptor integrin to a ligand (such as the extracellular matrix). This blocks integrin ligand binding and the transmission of signals transmitted to cells via integrin. The former inhibits cell adhesion, TGFβ activation, cell migration, cell migration, etc., while the latter inhibits cell differentiation and proliferation. Analyses such as, but not limited to, cell adhesion inhibition tests, cell detachment tests, and ligand binding inhibition of recombinant or purified soluble integrin heterodimers can be used to test whether a compound has the activity of inhibiting integrin function.

[0059] 3. Pharmaceutical Composition The pharmaceutical composition of the present invention is a composition comprising a compound represented by formula (I) or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier, and can be obtained, for example, by mixing a compound represented by formula (I) or a pharmaceutically acceptable salt thereof with a pharmaceutically acceptable carrier. The pharmaceutical composition of the present invention can treat or prevent various diseases by inhibiting integrin function as one of its mechanisms of action. The structure of the compound represented by formula (I) is as described in Section 1 above. The pharmaceutical composition of the present invention can be, for example, but is not limited to, tablets, granules, capsules, powders, liquids, injections, suppositories, patches, eye drops, and inhalants.

[0060] "Pharmaceutically acceptable salts" include, but are not limited to, sodium salts, potassium salts, ammonium salts, etc., when the compound has an acidic functional group. Furthermore, when the compound has a basic functional group, the salts can be salts with hydrochloric acid, phosphoric acid, acetic acid, phthalic acid, fumaric acid, oxalic acid, etc.

[0061] As the "pharmaceutically acceptable carrier" used in the pharmaceutical composition of the present invention, various inorganic or organic carrier substances can be used. When the pharmaceutical composition is prepared as a solid preparation such as a tablet or granule, excipients, lubricants, binders, disintegrants, etc. can be used. When the pharmaceutical composition is prepared as a liquid preparation such as a liquid preparation or an injection, solvents, solubilizers, suspending agents, buffers, etc. can be used. Furthermore, additives such as antioxidants, preservatives, colorants, etc. can also be used as needed.

[0062] Examples of excipients that can be used include, but are not limited to, lactose, D-mannitol, starch, etc., lubricants that can be used include magnesium stearate, talc, etc., binders that can be used include crystalline cellulose, gelatin, etc., and disintegrants that can be used include carboxymethyl cellulose, etc. Furthermore, examples of solvents that can be used include distilled water, alcohol, propylene glycol, etc., solubilizers that can be used include polyethylene glycol, ethanol, etc., suspending agents that can be used include stearyl triethanolamine, sodium lauryl sulfate, etc., and buffers that can be used include phosphates, acetates, etc.

[0063] 4. Therapeutic / Preventive Agent for Diseases Accompanying Fibrosis The therapeutic or preventive agent for diseases accompanied by fibrosis of the present invention contains, as an active ingredient, a compound represented by formula (I) or a pharmaceutically acceptable salt thereof. The structure of the compound represented by formula (I) is as described in Section 1 above, and its pharmaceutically acceptable salts are as described in Section 3 above. In the present invention, diseases accompanied by fibrosis are diseases accompanied by fibrosis of organs or tissues, and include pulmonary fibrosis, hepatic fibrosis, renal fibrosis, pancreatic fibrosis, scleroderma, keloid, skin cicatricialization, retinal cicatricialization, corneal cicatricialization, drug-induced gingival hyperplasia, frozen shoulder, arthritis, rheumatoid arthritis, and myocardial fibrosis that progresses in diabetic cardiomyopathy.

[0064] In the present invention, "renal fibrosis" includes diabetic nephropathy, chronic glomerulonephritis, and hypertensive nephrosclerosis. In the present invention, "pulmonary fibrosis" includes idiopathic interstitial pneumonia, collagen disease pneumonia, pneumoconiosis, drug-induced interstitial pneumonia, radiation pneumonitis, and pulmonary fibrosis associated with sarcoidosis and hypersensitivity pneumonitis, as well as acute exacerbation of any of the above interstitial pneumonias or pulmonary fibrosis, and acute interstitial pneumonia. In the present invention, "liver fibrosis" includes viral hepatitis, fatty liver, hepatitis associated with alcoholic hepatitis, hepatitis associated with non-alcoholic hepatitis (NASH), liver cirrhosis, liver fibrosis, and liver fibrosis associated with primary biliary cholangitis and autoimmune hepatitis.

[0065] The therapeutic or preventive agent for diseases accompanied by fibrosis of the present invention contains the compound represented by formula (I) or a pharmaceutically acceptable salt thereof as an active ingredient, and thereby exerts the effect of alleviating the symptoms of diseases accompanied by fibrosis, such as renal fibrosis, interstitial pneumonia, or liver fibrosis, or preventing the onset of diseases accompanied by fibrosis. Such efficacy is thought to be based on the mechanism of action of the compound represented by formula (I) or a pharmaceutically acceptable salt thereof, in which the compound represents integrin function.

[0066] The compound represented by formula (I) contained as an active ingredient in the therapeutic or preventive agent for diseases accompanied by fibrosis of the present invention can have a wide variety of chemical structures in combinations of A, B, X, and Y. Therefore, the chemical structure of the therapeutic or preventive agent for diseases accompanied by fibrosis of the present invention can be modified so as to optimize the absorbability, distribution, degradability, ease of excretion, etc. of the drug.

[0067] The therapeutic or preventive agent for diseases accompanied by fibrosis of the present invention can be administered as a therapeutic or preventive agent not only to patients diagnosed with diseases accompanied by fibrosis, but also to patients who may have these diseases or who are at risk of developing these diseases.

[0068] As described in Section 3 above, the therapeutic or preventive agent for diseases accompanied by fibrosis of the present invention can be mixed with a pharmaceutically acceptable carrier and formulated into various dosage forms. When used as a therapeutic or preventive agent for pulmonary fibrosis, it can be orally administered as tablets, granules, capsules, powders, liquids, etc., but can also be made into an inhalant or the like from the viewpoint of acting directly on the lungs to reduce side effects. Furthermore, depending on the symptoms, the effect can be enhanced by intravenous injection. When used as a therapeutic or preventive agent for liver fibrosis, the dosage form is not limited to these, but it can be orally administered as tablets, granules, capsules, powders, liquids, etc. Furthermore, from the viewpoint of acting directly on the liver to reduce side effects, it can also be administered directly to the liver as an injection via a tube or the like.

[0069] The route of administration of the therapeutic or preventive agent for fibrotic diseases of the present invention may be either oral or parenteral, with the daily dosage varying depending on the type of compound, the administration method, the patient's symptoms, age, and other factors. For example, oral administration typically involves administering approximately 0.01 to 1000 mg, more preferably approximately 0.1 to 500 mg, even more preferably approximately 0.1 to 50 mg, and even more preferably approximately 0.1 to 10 mg per kg body weight of a human or mammal, in one or several divided doses. Parenteral administration, such as intravenous injection, typically involves administering approximately 0.01 to 300 mg, more preferably approximately 1 to 100 mg, even more preferably approximately 1 to 50 mg, and even more preferably approximately 1 to 10 mg per kg body weight of a human or mammal. The compounds of the present invention have been confirmed to be effective even at extremely low oral doses, such as 1 mg / kg, allowing for reduced dosages compared to conventional methods. In this regard, conventional integrin inhibitor compounds typically require a dosage of 100 mg / kg, and even the lowest dosages are 50 mg / kg.

[0070] The therapeutic or preventive agent for diseases accompanied by fibrosis of the present invention may contain, in addition to the compound of the present invention or a pharmaceutically acceptable salt thereof, at least one active ingredient selected from drugs classified as therapeutic or preventive agents for diseases accompanied by fibrosis. Such active ingredients include, but are not limited to, for example, pirfenidone, nintedanib, etc. Furthermore, the therapeutic or preventive agent for diseases accompanied by fibrosis of the present invention can be used in combination with other therapeutic or preventive agents for diseases accompanied by fibrosis.

[0071] The present invention will be described in detail below with reference to examples, but the present invention is not limited to these examples.

[0072] Example 1 Synthesis of IBF-17 IBF-17, one of the compounds of the present invention, was produced. First, (R,E)-2-methyl-N-[3-(pentafluorosulfanyl)benzylidene]propane-2-sulfinamide (intermediate 1-1) represented by the following structural formula was synthesized by the following steps.

[0073]

[0074] To 7.52 g of 3-(pentafluorosulfanyl)benzaldehyde, 75 ml of dehydrated toluene, 4.12 g of (R)-2-methylpropane-2-sulfinamide, 0.130 ml of pyrrolidine, and 32 g of molecular sieves (4 angstroms) were added in a room temperature atmosphere, and the mixture was stirred at 70°C for 3 hours under an argon atmosphere. 0.130 ml of pyrrolidine was then added at room temperature, and the mixture was stirred at 70°C for 6 hours. After completion of the reaction, the reaction solution was filtered through Celite and washed with 125 ml of toluene. The filtrate and washings were concentrated under reduced pressure, n-hexane was added, the solid was crushed, ultrasonically treated, and the mixture was stirred for a while in an ice bath. The solid was collected by filtration, washed with cold n-hexane, and dried under reduced pressure to obtain 7.50 g of Intermediate 1-1 as a pale yellow solid. The filtrate and washings were concentrated under reduced pressure, n-hexane was added, the solid was crushed, and the mixture was subjected to ultrasonic treatment. The solid was then filtered, washed with cold n-hexane, and dried under reduced pressure to obtain 0.82 g of Intermediate 1-1 as a white solid. The results of mass analysis and NMR measurement spectrum of Intermediate 1-1 are as follows. Mass spectrum (DUIS, m / z): 336 [M+H] + 1 H-NMR(400MHz, CDCl3)δ: 8.62(s, 1H), 8.27-8.23(m, 1H), 8.01-7.96(m, 1H), 7.93-7.88(m, 1H), 7.61(t, J=7.9Hz, 1H), 1.29(s, 9H)

[0075] Subsequently, methyl (S)-3-[((R)-tert-butylsulfinyl)amino]-3-[3-(pentafluorosulfanyl)phenyl]propanoate (intermediate 1-2) represented by the following structural formula was synthesized as follows.

[0076]

[0077] 2.47 g of copper(I) chloride was added to 16.20 g of zinc, and the mixture was degassed under reduced pressure at 70-80°C for 15 minutes. The mixture was purged with argon, and 80 ml of dehydrated tetrahydrofuran was added under an argon atmosphere, followed by stirring at 70°C for 30 minutes. The mixture was then cooled to 50°C, and 5.80 ml of methyl bromoacetate was added in portions so that the internal temperature did not exceed 60°C, followed by stirring at 50°C for 1 hour. After returning to room temperature, the mixture was cooled in an ice-salt bath, and a solution of 8.32 g of (R,E)-2-methyl-N-[3-(pentafluorosulfanyl)benzylidene]propane-2-sulfinamide (intermediate 1-1) in 40 ml of dehydrated tetrahydrofuran was added dropwise under an argon atmosphere at an internal temperature of -10°C or below. The mixture was then washed in with 10 ml of dehydrated tetrahydrofuran and stirred in an ice bath for 2.5 hours. After completion of the reaction, the reaction solution was filtered through Celite and washed with 130 ml of tert-butyl methyl ether. 110 ml of 10% aqueous citric acid was added to the filtrate and washings, and the mixture was stirred at room temperature for a while, followed by separation. The organic layer was washed successively with 70 ml of saturated saline, 70 ml of saturated sodium bicarbonate water, and 70 ml of saturated saline, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting concentrated residue was purified by column chromatography (silica gel, elution solvent: 1,2-dichloroethane: ethyl acetate) to obtain 8.89 g of Intermediate 1-2 as a white solid. The results of mass analysis and NMR measurement spectrum of Intermediate 1-2 are as follows. Mass spectrum (DUIS, m / z): 410 [M+H] + 1 H-NMR (400MHz, DMSO-d6) δ:7.92-7.88(m, 1H), 7.82-7.77(m, 1H), 7.69-7.64(m, 1H), 7.61-7.54 (m, 1H), 5.80(d, J=6. 1Hz, 1H), 4.82-4.75(m, 1H). 3.55(s, 3H), 3.04 (dd, J=7.2, 15.7Hz, 1H), 2.84(dd, J=7.3, 15.7Hz, 1H), 1.07(s, 9H).

[0078] Furthermore, methyl (S)-3-amino-3-[3-(pentafluorosulfanyl)phenyl]propanoate hydrochloride (intermediate 1-3) represented by the following structural formula was synthesized as follows.

[0079]

[0080] 90 ml of dehydrated diethyl ether, 0.86 ml of dehydrated methanol, and 47.0 ml of a 1M hydrogen chloride diethyl ether solution were added to 8.66 g of methyl (S)-3-[((R)-tert-butylsulfinyl)amino]-3-[3-(pentafluorosulfanyl)phenyl]propanoate (intermediate 1-2) at room temperature, and the mixture was stirred at room temperature for 1 hour under an argon atmosphere. After completion of the reaction, the reaction solution was ice-cooled, and the precipitated solid was collected by filtration, washed with dehydrated diethyl ether, and then dried under reduced pressure to obtain 7.10 g of intermediate 1-3 as a white solid. The results of mass analysis and NMR measurement spectrum of intermediate 1-3 are as follows. Mass spectrum (DUIS, m / z): 306 [M+H] + 1 H-NMR (400MHz, DMSO-d6) δ:8.82(br s, 3H), 8.18-8.14(m, 1H), 7.96-7.87(m, 2H), 7.72-7.65(m, 1H), 4.79 (dd, J=6.0, 8.5Hz, 1H), 3.56(s, 3H), 3.24(dd, J=6.0, 16.5Hz, 1H), 3.08 (dd, J=8. 5, 16. 5Hz, 1H).

[0081] In addition, 2-fluoromalonamide (intermediate 1-9) represented by the following structural formula was synthesized as follows.

[0082]

[0083] To a solution of 25.0 g of diethyl 2-fluoromalonate in methanol (100 ml) was added 80 ml of 7N ammonia methanol solution at room temperature under an argon stream, and the mixture was stirred at room temperature for 4 hours. After completion of the reaction, the solid was collected by filtration, washed with methanol, and then dried under reduced pressure at 50°C to obtain 15.91 g of Intermediate 1-9 as a white solid. The NMR spectrum of Intermediate 1-99 is shown below. 1 H-NMR (400MHz, DMSO-d6) δ:7.78-7.48(m, 4H), 5.16(d, J=48.9Hz, 1H).

[0084] Then, 2-fluoropropane-1,3-diamine hydrochloride (intermediate 1-10) represented by the following structural formula was synthesized as follows.

[0085]

[0086] To 400 ml of a 0.9 M borane-tetrahydrofuran solution, 7.20 g of 2-fluoromalonamide (Intermediate 1-9) was added in portions at room temperature under an argon stream, and the mixture was stirred at room temperature for 20 hours. After completion of the reaction, the reaction solution was cooled to 0°C, and 73 ml of methanol was added dropwise at 20°C or below. The mixture was stirred at room temperature for 1 hour and then concentrated under reduced pressure. 40 ml of ethanol was added to the concentrated residue, and 60 ml of a 2 M hydrogen chloride ethanol solution was added at 0°C, and the mixture was stirred at room temperature for 3 hours. The precipitated solid was collected by filtration, dissolved in methanol, and concentrated under reduced pressure. 70 g of Amberlite (IRA478RF CL), which had been washed successively with 200 ml of water and 200 ml of methanol, was added at room temperature to a solution of 6.91 g of the resulting solid in 200 ml of methanol, and the mixture was stirred at room temperature for 30 minutes. The reaction solution was filtered, and the filtrate was concentrated under reduced pressure to obtain 4.55 g of Intermediate 1-10 as a white solid. The NMR spectrum of Intermediate 1-10 is as follows: 1 H-NMR (400MHz, DMSO-d6+D2O) δ: 4.77-4.53(m, 1H), 3.01-2.79 (m, 4H).

[0087] Furthermore, 3-[(5-fluoro-1,4,5,6-tetrahydropyrimidin-2-yl)amino]-5-hydroxybenzoic acid (intermediate 1-6) represented by the following structural formula was synthesized as follows.

[0088]

[0089] To a solution of 1.0 g of 3-amino-5-hydroxybenzoic acid in 6 ml of dehydrated N,N-dimethylformamide, a solution of 0.60 g of methyl isothiocyanate in 6 ml of dehydrated N,N-dimethylformamide was added at room temperature under an argon atmosphere, and the mixture was stirred at room temperature for 30 minutes, followed by stirring at 60°C for 17.5 hours. The reaction mixture was then cooled to room temperature, and 1.0 ml of iodomethane was added in an ice bath. The mixture was stirred at room temperature for 1.5 hours, and the reaction mixture was concentrated under reduced pressure. To the resulting concentrated residue, 2.09 g of 2-fluoropropane-1,3-diamine hydrochloride (intermediate 1-10) and 10 ml of dehydrated N,N-dimethylformamide were added in an ice bath, and the mixture was stirred at 50°C for 30 minutes, followed by stirring at 95°C for 4.5 hours. After completion of the reaction, the reaction mixture was cooled to room temperature and added to 10 ml of water. 6N hydrochloric acid was added in an ice bath to adjust the pH to 6.0. The solvent was partially removed by concentration under reduced pressure, and the mixture was stirred overnight at room temperature. The precipitated solid was collected by filtration, washed successively with water and acetonitrile, and then dried under reduced pressure to obtain 356 mg of Intermediate 1-6 as a white solid. The results of mass analysis and NMR measurement spectrum of Intermediate 1-6 are as follows. Mass spectrum (ESI, m / z): 254 [M+H] + 1 H-NMR (400MHz, DMSO-d6+D2O) δ: 7.19-7.13(m, 2H), 6.76-6.70(m, 1H), 5.43-5.17(m, 1H), 3.73-3.40(m, 4H).

[0090] Subsequently, {3-[(5-fluoro-1,4,5,6-tetrahydropyrimidin-2-yl)amino]-5-hydroxybenzoyl}glycine (intermediate 1-7) represented by the following structural formula was synthesized as follows.

[0091]

[0092] To a suspension of 300 mg of 3-[(5-fluoro-1,4,5,6-tetrahydropyrimidin-2-yl)amino]-5-hydroxybenzoic acid (intermediate 1-6) in N,N-dimethylformamide (1.5 ml)-dichloromethane (1.5 ml), 182 mg of ethyl glycinate hydrochloride and 0.22 ml of N,N'-diisopropylcarbodiimide were added at room temperature under an argon stream, and the mixture was stirred at room temperature for 19 hours. Thereafter, 3.6 ml of 2N aqueous sodium hydroxide solution was added, and the mixture was stirred at room temperature for 1.5 hours. After completion of the reaction, dichloromethane was added to the reaction solution and the layers were separated. 6N hydrochloric acid was added to the aqueous layer to adjust the pH to 5.0, and the mixture was stirred at room temperature for 2 hours. The precipitated solid was collected by filtration, washed successively with water and acetonitrile, and then dried at 60°C under reduced pressure to obtain 188 mg of intermediate 1-7 as a white solid.

[0093] Thereafter, methyl (3S)-3-(2-{3-[(5-fluoro-1,4,5,6-tetrahydropyrimidin-2-yl)amino]-5-hydroxybenzamido)acetamido)-3-[3-(pentafluorosulfanyl)phenyl]propanoate trifluoroacetate (intermediate 1-8) represented by the following structural formula was synthesized as follows.

[0094]

[0095] To a suspension of 300 mg of {3-[(5-fluoro-1,4,5,6-tetrahydropyrimidin-2-yl)amino]-5-hydroxybenzoyl}glycine (Intermediate 1-7) in N,N-dimethylformamide (3 ml), 363 mg of methyl (S)-3-amino-3-[3-(pentafluorosulfanyl)phenyl]propanoate hydrochloride (Intermediate 1-3) and 144 mg of 1-hydroxybenzotriazole were added at room temperature under an argon stream, and the mixture was stirred at room temperature for 10 minutes. Next, 0.18 ml of N,N'-diisopropylcarbodiimide was added at room temperature, and the mixture was stirred at room temperature for 15 hours. After completion of the reaction, the reaction solution was purified by silica gel chromatography (silica gel, elution solvent: 0.1% TFA acetonitrile: 0.1% TFA aqueous solution) to obtain 673 mg of Intermediate 1-8 as a pale yellow solid. The results of mass spectrometry and NMR measurement spectrum of Intermediate 1-8 are as follows. Mass spectrum (ESI, m / z): 598[M+H] + 1 H-NMR (400MHz, DMSO-d6) δ: 10.05(br s,1H), 9.78(s, 1H), 8.69-8.63(m, 2H), 8.28(br s, 2H), 7.87-7.84(m, 1H), 7.82-7.76(m, 1H), 7.68-7.63(m, 1H), 7.62-7.56(m, 1H), 7.17-7.14(m, 1H), 7.14-7.11(m, 1H), 6.75(t, J=2. 0Hz, 1H), 5.38-5.18(m, 2H), 3.94-3.79(m, 2H), 3.60-3.39(m, 7H), 2.86(d, J=7.4Hz, 2H).

[0096] Finally, (3S)-3-(2-{3-[(5-fluoro-1,4,5,6-tetrahydropyrimidin-2-yl)amino]-5-hydroxybenzamido}acetamido)-3-[3-(pentafluorosulfanyl)phenyl]propanoic acid (IBF-17), represented by the following structural formula, was synthesized as follows.

[0097]

[0098] To a solution of 670 mg of methyl (3S)-3-(2-{3-[(5-fluoro-1,4,5,6-tetrahydropyrimidin-2-yl)amino]-5-hydroxybenzamido}acetamido)-3-[3-(pentafluorosulfanyl)phenyl]propanoate trifluoroacetate (Intermediate 1-8) in acetonitrile (3.5 ml) / water (3.5 ml), 113 mg of lithium hydroxide was added at room temperature under an argon stream, and the mixture was stirred at room temperature for 2 hours. After completion of the reaction, 1N hydrochloric acid was added to the reaction solution to adjust the pH to 5.8. After filtration, the filtrate was concentrated under reduced pressure, the acetonitrile was distilled off under reduced pressure, and the mixture was stirred at room temperature for 17 hours. A small amount of acetonitrile was added to obtain a homogeneous solution, which was then purified with BondElut (elution solvent: water:acetonitrile:methanol=100:0:0 to 0:100:0 to 0:67:33) to obtain 426 mg of IBF-17 as a light brown solid. The results of mass spectrometry and NMR measurement spectrum of IBF-17 are as follows. Mass spectrum (ESI, m / z): 584 [M+H] + 1 H-NMR (400MHz, CD3OD) δ:7.83 (t, J=l.8Hz, 1H), 7.68-7.58(m, 2H), 7.51-7.42(m, 1H), 7.23-7.20(m, 1H), 7.20-7.17(m, 1H), 6.78(t, J=2.1Hz, 1H), 5.28(t, J=5.5Hz, 1H), 5.25-5.08(m, 1H), 4.09(d, J=16.9Hz, 1H), 4.01(d, J=16.9Hz, 1H),3.70-3.51(m, 3H), 3.50-3.40(m, 1H), 2.70(dd, J=5.5, 15.3Hz, 1H), 2.63(dd, J=5. 5, 15.3Hz, 1H).

[0099] Example 2: Effect on TGFβ activation In this example, lung fibroblasts were used as integrin-expressing cells to confirm the effect of IBF-17, a compound of the present invention, on TGFβ activity. Fibroblasts isolated from male Wistar rat lungs and cultured for 7 days were placed at 2 × 10 per well. ３After 5 days, IBF-17 or C8 (known αvβ1 inhibitor) was added to a final concentration of 2, 20, or 200 nM, and 2 hours later, 2×10 TGFβ-responsive luciferase-producing reporter cells CCL64 were added to the cells. ４ The cells were seeded with αvβ1 and co-cultured for 18 hours. After incubation, the cells were lysed, luciferase assay reagent (Promega) was added, and luciferase activity was measured using a luminometer. The results are shown in Figure 1. The structure of C8, a known αvβ1 inhibitor, is as follows:

[0100]

[0101] For comparison, Figure 1 shows the results for reporter cells alone on the far left, and the results for a mixed culture of reporter cells and lung fibroblasts on the right. The results for IBF17 and C8 at 2, 20, and 200 nM are also shown on the right. The results of this experiment showed that IBF17 concentration-dependently inhibited TGFβ activation over the range of 2-200 nM. At 200 nM, luciferase activity was close to that of reporter cells alone, demonstrating nearly complete inhibition of TGFβ activation by integrins expressed in lung fibroblasts. In contrast, C8, an αvβ1-selective inhibitor, showed no inhibition at 2 nM, but inhibited TGFβ activation at 20 nM and 200 nM, but the inhibitory effect was significantly different from that of IBF17 (IBF17 had a significantly stronger inhibitory effect).

[0102] Example 3: Effect on myofibroblast differentiation of fibroblasts. In this example, the effect of IBF-17, a compound of the present invention, on myofibroblast differentiation of fibroblasts was examined. The medium was replaced with serum-free medium, and the rat lung fibroblasts isolated in Example 2 were cultured for 24 hours. After culture, IBF-17 was added to a final concentration of 2, 20 nM, or 200 nM, and C8 was added to a final concentration of 2, 20, or 200 nM, and the cells were incubated for 2 hours. After incubation, recombinant human TGF-β1 was added to a final concentration of 5 ng / ml, and the cells were stimulated for 96 hours to induce differentiation into myofibroblasts. After differentiation induction, proteins were extracted from the cells and subjected to Western blotting to detect the bands of the differentiation marker α-SMA. The band intensities were quantified as the relative band intensity to that of the control glyceraldehyde-3-phosphate dehydrogenase (GAPDH).

[0103] Figure 2 shows the results of Western blotting using anti-α-smooth muscle actin antibody (α-SMA) and anti-GAPDH antibody. Figure 3 shows the quantification of band intensities, expressed relative to the band intensity of cells to which TGFβ was not added. In this example, α-smooth muscle actin was used as an indicator to evaluate the differentiation of fibroblasts into myofibroblasts (by 0.2 ng / ml TGFβ). In the case of the addition of IBF17 shown in Figure 3(A), differentiation inhibition was observed at a concentration of 2 nM, and at a concentration of 20 nM, inhibition was 80% or more. Meanwhile, Figure 3(B) similarly observes the effect of the addition of C8, but there was no effect at concentrations of 2 and 20 nM, and inhibition was finally observed at 200 nM.

[0104] C8, which was used as a control for IBF17 in Examples 2 and 3, had lower effects than IBF17 in TGFβ activation and myofibroblast differentiation, which are indicators for estimating fibrotic activity in vitro. However, since administration of C8 alone suppresses in vivo fibrosis in the liver, lung, kidney, and heart (The αvβ1 integrin plays a critical in vivo role in tissue fibrosis, Nilgun I Reed, Hyunil Jo, Chun Chen, Kazuyuki Tsujino, Thomas D Arnold, William F DeGrado, Dean Sheppard. Sci Transl Med 20; 8(288): 288ra79, 2015), IBF17 is expected to have even greater in vivo effects.

[0105] (Example 4-1) Carbon tetrachloride (CCl ４ In this example, mice were administered carbon tetrachloride (CCl ) for 8 weeks to induce liver fibrosis. ４ ) and compounds of the present invention (IBF-17, IBF-18, and IBF-20) were administered, and the state of tissue fibrosis was observed. Mice were intraperitoneally administered carbon tetrachloride (diluted 4-fold with olive oil) at 1 ml / kg twice per week (e.g., Monday, Friday, or Tuesday, Friday) for a total of 8 weeks. IBF-17, IBF-18, or IBF-20 was orally administered at 1 or 10 mg / kg twice per day (with an 8-hour interval) throughout the entire study period. After the experiment, livers were removed and collagen content was quantified by hydroxyproline measurement. Furthermore, the specimens were stained with Sirius Red to quantify the area of ​​fibrosis. In addition, the expression of fibrosis markers, Col1a1 and Acta2 (α-SMA), was analyzed by qPCR.

[0106] Figure 4 shows the dosing schedule, Figure 5 shows the results of quantification of hydroxyproline in liver tissue, Figure 6 shows a comparison of Sirius Red-stained tissue areas and photographs of the tissues after staining, and Figure 7 shows the mRNA expression levels (relative intensity) of fibrosis marker genes (Col1a1 gene and Acta2 gene). Figure 5 confirms that administration of IBF-17 and IBF-18 reduced hydroxyproline levels in mice, but IBF-20 did not show this effect at a dose of 10 mg / kg. When comparing IBF-17 and IBF-18, which were effective, only IBF-17 showed an effect at 1 mg / kg. Therefore, we examined the reduction in Sirius Red-stained liver tissue area and fibrosis markers in the IBF-17-treated group. In both the 1 mg / kg and 10 mg / kg IBF-17 groups, the area stained with Sirius Red was reduced compared to the control group, and the degree of reduction corresponded to the dose. The right side of Figure 6 shows examples of staining, and sufficient staining is observed in all cases. Figure 7 indicates a tendency for IBF-17 to reduce the expression of both Col1a1 and Acta2.

[0107] (Example 4-2) Carbon tetrachloride (CCl ４ In this example, mice were administered carbon tetrachloride (CCl) to induce liver fibrosis. ４ ) was administered once, followed by administration of the compound of the present invention (IBF-17) for three days, after which the livers were excised and observed. More specifically, mice were fasted for half a day from the day before the test and then intraperitoneally administered 2 ml / kg of carbon tetrachloride (diluted 4-fold with olive oil) once. IBF-17 was administered subcutaneously at 1 mg / kg twice daily (with an 8-hour interval), including the day of carbon tetrachloride administration. 72 hours after carbon tetrachloride administration, the mice were dissected, and the livers were excised. The expression of fibrosis marker genes (Acta2, Col1a1, Col3a1, Tgfb1, Loxl1, and Ddr2) was analyzed by qPCR.

[0108] Figure 8 shows the dosing schedule, and Figure 9 shows the mRNA expression level (relative intensity) by qPCR. Figure 9 shows the analysis results for six types of fibrosis markers. ４It can be seen that all markers increased after 72 hours of treatment. However, IBF-17 administration appeared to decrease all of these markers, with a clear decrease in the levels of two collagen species, Col1a1 and Col3a1.

[0109] Example 5: 8-Week Administration Experiment in a NASH Model In this example, mice were fed a CDAHFD (a choline-deficient, amino-restricted, high-fat diet) for 8 weeks while receiving the compounds of the present invention (IBF-17, IBF-18, IBF-20), and the state of tissue fibrosis was observed. More specifically, mice were fed a special diet, CDAHFD (Research Diets), which induces human NASH-like pathology, with fresh food replaced twice a week (e.g., Monday and Friday, or Tuesday and Friday) for a total of 8 weeks. IBF-17, IBF-18, or IBF-20 was orally administered twice daily (with an 8-hour interval) at 1, 3, or 10 mg / kg throughout the entire study period. After the experiment, the amount of collagen in the liver was quantified by hydroxyproline measurement. For IBF-17, specimens were stained with Sirius Red, which specifically stains collagen, to quantify the area of ​​fibrosis. In addition, the expression of genes considered to be fibrosis markers (Acta2, Col1a1, and Timp-1) was analyzed by qPCR. During the feeding period, the body weight of each mouse (which temporarily decreases in the early stages of feeding in CDAHFD) was measured, and the weight of the excised liver was also measured.

[0110] Figure 10 shows the dosing schedule, Figure 11 shows the quantification of hydroxyproline in liver tissue (p<0.001), Figures 12(A) and (B) show changes in mouse body weight and liver weight after 56 weeks, Figure 13 shows a comparison of Sirius Red-stained tissue areas and photographs of the tissues after staining, and Figure 14 shows the mRNA expression levels (relative intensity) of genes considered to be fibrosis markers (Acta2 gene, Col1a1 gene, and Timp-1 gene). Figure 11 shows that for all compounds (IBF-17, IBF-18, IBF-20), at all doses (1, 3, and 10 mg / kg), a decrease in hydroxyproline was observed compared to the control group. The CDAHFD diet used to generate NASH model mice induces fatty deposits in the liver, and after administration, the liver becomes enlarged and increases in weight. Figure 12 shows liver weights at the end of the experiment. CDAHFD administration increased liver weights by more than 1.5 times that of the normal diet group, but this increase was similar for all drug doses (1, 3, and 10 mg / kg), indicating that the drugs had no effect on liver weight. Figures 13 and 14 show that IBF-17 reduced the fibrotic area in tissue specimens at doses of 1, 3, and 10 mg / kg compared to the untreated group, and consistently reduced fibrosis markers. It is noteworthy that even a dose of just 1 mg / kg produced the same effect as doses of 3 and 10 mg / kg.

[0111] Example 6: 10-Day Administration Experiment in a Unilateral Ureteral Obstruction (UUO) Model In this example, mice with one ureter ligated were administered the compound of the present invention (IBF-17) for 10 days, and the state of tissue fibrosis was observed. More specifically, mice under deep anesthesia underwent abdominal surgery, and the left ureter was ligated at two locations using nylon thread, and the abdomen was sutured. Starting one day after ligation, IBF-17 was administered subcutaneously at 1 or 5 mg / kg twice daily (with an 8-hour interval) for nine days. Ten days after ligation, the kidneys were removed, and the expression of genes considered to be fibrosis markers (Acta2, Col1a1, Col3a1, Ddr2, Tgfb1, Tnc, Trpc6, and Loxl1) was analyzed by qPCR.

[0112] Figure 15 shows the dosing schedule, and Figure 16 shows the mRNA expression levels (relative intensity) measured by qPCR. Of the eight fibrosis markers, Acta2, Tgfbl, Tnc, and Trpc6 all showed a tendency to decrease, while Col1a1, Col3a1, Ddr2, and Loxl1 showed significant decreases. This example confirmed that IBF17 is also effective against renal fibrosis.

[0113] Example 7: Study of effective dose and no-toxic dose Figure 17 shows the area under the blood concentration-time curve (AUC) when the compounds of the present invention (IBF-17, IBF-18, IBF-20) were administered at doses of 1, 10, 30, 100, and 300 mg / kg, and the margin comparing the effective dose (1 mg / kg) and the no-toxic dose (no-observable adverse effect level: NOAE). From these results, it is expected that the compounds of the present invention have a very large margin (PK / TK) between the effective dose and the no-toxic dose (toxic dose), and that the risk of side effects is extremely low.

[0114] (Example 8) Carbon tetrachloride (CCl ４ In this example, mice were administered carbon tetrachloride (CCl ) for 6 weeks to induce liver fibrosis. ４) and over the final three weeks of the study, a compound of the present invention (IBF-17) was administered for a total of 15 days, and the state of tissue fibrosis was observed. While Example 4 examined the preventive effect of administering a compound from the start of the study, this example examines the therapeutic effect of administering a compound three weeks after carbon tetrachloride administration (when liver fibrosis had occurred). Specifically, mice were intraperitoneally administered 1 ml / kg of carbon tetrachloride (diluted four-fold with olive oil) twice per week (e.g., Monday, Friday, or Tuesday, Friday) for a total of six weeks. Starting four weeks after the start of the study, IBF-17 was orally administered at 5 mg / kg twice per day (with an 8-hour interval), five days per week (Monday to Friday). After the experiment, the livers were removed and collagen levels were quantified by hydroxyproline measurement. Figure 18 shows the dosing schedule, and Figure 19 shows the results of hydroxyproline quantification in liver tissue. IBF-17 significantly reduced the amount of hydroxyproline in the mouse liver, and this test confirmed that IBF-17 has a therapeutic effect on liver fibrosis.

[0115] Example 9: 10-Day Administration Experiment in a Bleomycin-Administered Model. In this example, mice were administered bleomycin to induce pulmonary fibrosis. Starting one week after bleomycin administration, the compound of the present invention (IBF-17) was administered for a total of 10 days over a two-week period, beginning one week after bleomycin administration. The state of tissue fibrosis was then observed. Specifically, the neck of each mouse was incised under deep anesthesia, and bleomycin (diluted in saline) was administered intratracheally once at 1.5 U / kg. The neck was then sutured. Starting one week after bleomycin administration (when fibrosis begins), IBF-17 was orally administered at 5 mg / kg twice daily (with an 8-hour interval), five days a week (Monday to Friday). After the experiment, the lungs were excised and the expression of fibrosis-related genes (Acta2, Col1a1, Col3a1, Tnc) was analyzed by qPCR. Lung sections were also prepared and stained for collagen with Masson's trichrome staining, and the area of ​​fibrosis was quantified. Figure 20 shows the dosing schedule, Figure 21 shows the qPCR results, and Figure 22 shows the fibrosis area quantification results and photographs of the lungs after staining (completely joined photographs of the left lobe). Among the fibrosis-related genes, the expression of Col1a1 and Col3a1 was significantly reduced by IBF-17 administration. Furthermore, the fibrosis area was also significantly reduced by IBF-17 administration. These results confirmed that IBF-17 has a therapeutic effect on pulmonary fibrosis.

[0116] The above detailed description is merely illustrative of the objects and scope of the present invention and is not intended to limit the scope of the appended claims. Various modifications and substitutions to the described embodiments will be apparent to those skilled in the art from the teachings set forth herein, without departing from the scope of the appended claims.

[0117] This application claims priority based on Japanese Patent Application No. 2023-102623 (filing date: June 22, 2023), the contents of which are incorporated herein by reference. The entire contents of Japanese Patent Application No. 2023-102623 are incorporated herein by reference and form a part of this specification.

[0118] The present invention provides novel compounds that inhibit the function of integrins, which are useful for treating or preventing diseases associated with fibrosis.

Claims

1. Formula (I) (Wherein, A is C—OH or N, and B is C—H, C—COOH, C—CH ３ , C—Br, or N; X is —SF ５ , or -Si(CH ３ ) ３ Y is -F or -OH, and * represents an asymmetric carbon atom, or a salt thereof.

2. X is -SF ５ B is C-H, C-CH ３ or C--Br, or a salt thereof.

3. The compound or salt thereof according to claim 2, wherein Y is -F.

4. The compound or salt thereof according to claim 3, wherein A is C-OH.

5. X is -Si(CH ３ ) ３ The compound or salt thereof according to claim 1, wherein B is C-COOH or N.

6. An integrin inhibitor comprising the compound according to any one of claims 1 to 5 or a pharma- ceutically acceptable salt thereof.

7. A pharmaceutical composition comprising a compound according to any one of claims 1 to 5 or a pharma- ceutically acceptable salt thereof and a pharma- ceutically acceptable carrier.

8. A therapeutic or preventive agent for a disease accompanied by fibrosis, comprising as an active ingredient the compound according to any one of claims 1 to 5 or a pharma- ceutically acceptable salt thereof.

9. The therapeutic or preventive agent for a disease accompanied by fibrosis according to claim 8, wherein the disease accompanied by fibrosis is kidney fibrosis, lung fibrosis, or liver fibrosis.

10. The therapeutic or preventive agent for diseases accompanied by fibrosis according to claim 8, for treating or preventing a disease accompanied by fibrosis in combination with at least one or more drugs selected from drugs classified as therapeutic or preventive agents for diseases accompanied by fibrosis.

11. A therapeutic or preventive agent for diseases accompanied by fibrosis, comprising a combination of the compound according to any one of claims 1 to 5 or a pharma- ceutically acceptable salt thereof and at least one active ingredient selected from drugs classified as therapeutic or preventive agents for diseases accompanied by fibrosis.

12. The compound according to any one of claims 1 to 5 or a pharma- ceutically acceptable salt thereof for use as a therapeutic or prophylactic agent for a disease associated with fibrosis.

13. Use of a compound according to any one of claims 1 to 5 or a pharma- ceutically acceptable salt thereof for the manufacture of a medicament for treating or preventing a disease associated with fibrosis.

14. A method for treating or preventing a disease accompanied by fibrosis, comprising administering to a patient a compound according to any one of claims 1 to 5 or a pharma- ceutically acceptable salt thereof.

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