Compound pharmaceutical preparation containing oil component comprising polyoxyethylene structure

Abstract

The present invention relates to a pharmaceutical preparation containing a compound and an oily component comprising a polyoxyethylene structure. The content of oil component comprising a polyoxyethylene structure is 2.0 to 7.5 vol% in 100 vol% of the pharmaceutical preparation.

Description

Compound preparation containing an oily component containing a polyoxyethylene structure

[0001] The present invention relates to a compound preparation containing an oily component having a polyoxyethylene structure. The present invention also relates to a method for screening candidate compounds using the preparation.

[0002] In general, compounds with a molecular weight of 500 or more have low membrane permeability when orally administered, and are considered to have problems with oral absorption (e.g., Non-Patent Documents 1 and 2). Furthermore, compounds with low solubility and low membrane permeability tend to have low oral absorption. These compounds often fail to achieve sufficient absorption even when the dose is increased, making appropriate efficacy and safety evaluation impossible. For this reason, in pharmaceutical development, the selection of compounds with such properties has been avoided as much as possible during the screening stage.

[0003] On the other hand, when a compound with low oral absorbability is selected as a candidate compound for a pharmaceutical product, efforts have been made to improve oral absorbability by formulation techniques or drug delivery techniques (e.g., Non-Patent Document 3).

[0004] Donovan, M. D. et al. , Pharm. Res. , 1990, 7, pp. 863-868. Lipinski, C. A. , Adv. Drug Del. Rev. , 1997, 23, pp3-25. (Lipinski, rule of 5) Ghadi, R. et al. , J. Control. Release, 2017, 248, pp. 71-95.

[0005] However, the optimal additive and / or formulation may differ for each compound, and in the early stage of screening candidate compounds when there are a large number of test compounds, selecting the optimal additive and / or formulation for each test compound and administering each test compound with a different additive and / or formulation is not realistic, since it would require optimizing different additives and / or formulations for each test compound.

[0006] Furthermore, when oral administration animal testing is performed in the early stages of drug screening, test compounds are evaluated using formulations prepared using certain common additives, but depending on the compatibility between the test compound and the additive, sufficient oral absorption may not be achieved, and the test compound may not be evaluated appropriately. As a result, the screening process for candidate compounds is influenced by chance factors, and there is a possibility that useful test compounds may be overlooked as false negatives.

[0007] The present invention has been made in view of the above circumstances, and aims to provide a versatile formulation with improved oral absorbability that enables evaluation of compounds under the same formulation conditions. Another aim of the present invention is to provide a method for screening candidate compounds using the formulation.

[0008] One aspect of the present invention relates to a formulation. Specifically, for example, the present invention includes the following inventions. [A1] A formulation comprising a compound and an oily component containing a polyoxyethylene structure, wherein the content of the oily component in 100% by volume of the formulation is 2.0% by volume or more and 7.5% by volume or less. [A2] An absorption enhancer for a compound, comprising an oily component containing a polyoxyethylene structure as an active ingredient, wherein the content of the oily component in 100% by volume of the formulation containing the compound is 2.0% by volume or more and 7.5% by volume or less. [A3] A solubility enhancer for a compound, comprising an oily component containing a polyoxyethylene structure as an active ingredient, wherein the content of the oily component in 100% by volume of the formulation containing the compound is 2.0% by volume or more and 7.5% by volume or less. [A4] The formulation or agent according to any of [A1] to [A3], wherein the ClogP of the compound is 4 or more and 25 or less. [A5] The formulation or agent according to any one of [A1] to [A4], wherein the ClogP of the compound is 8 or more and 21 or less. [A6] The formulation or agent according to any one of [A1] to [A5], wherein the ClogP of the compound is 9 or more and 20 or less. [A7] The formulation or agent according to any one of [A1] to [A6], wherein the ClogP of the compound is 13 or more and 17 or less. [A8] The formulation or agent according to any one of [A1] to [A7], wherein the ClogP of the compound is 13 or more. [A9] The formulation or agent according to any one of [A1] to [A8], wherein the ClogP of the compound is equal to or greater than the ClogP of Compound 2. [A10] The formulation or agent according to any one of [A1] to [A9], wherein the ClogP of the compound is equal to or greater than the ClogP of Compound 2 and equal to or less than the ClogP of Compound 1. [A11] The formulation or agent according to any one of [A1] to [A10], wherein the compound has a Caco-2 Papp (cm / sec) value of 1.0E-9 or more. [A12] The formulation or agent according to any one of [A1] to [A11], wherein the compound has a Caco-2 Papp (cm / sec) value of 1.0E-7 or more.[A13] The formulation or agent according to any one of [A1] to [A12], wherein the Caco-2 Papp (cm / sec) value of the compound is 1.0E-4 or less. [A14] The formulation or agent according to any one of [A1] to [A13], wherein the Caco-2 Papp (cm / sec) value of the compound is 1.0E-5 or less. [A15] The formulation or agent according to any one of [A1] to [A14], wherein the Caco-2 Papp (cm / sec) value of the compound is 6.0E-6 or less. [A16] The formulation or agent according to any one of [A1] to [A15], wherein the Caco-2 Papp (cm / sec) value of the compound is 1.0E-9 or more and 1.0E-4 or less. [A17] The formulation or agent according to any one of [A1] to [A16], wherein the Caco-2 Papp (cm / sec) value of the compound is 1.0E-8 or more and 1.0E-5 or less. [A18] The formulation or agent according to any one of [A1] to [A17], wherein the Caco-2 Papp (cm / sec) value of the compound is 1.0E-7 or more and 1.0E-5 or less. [A19] The formulation or agent according to any one of [A1] to [A18], wherein the Caco-2 Papp (cm / sec) value of the compound is equal to or greater than the Papp (cm / sec) value of Compound 9. [A20] The formulation or agent according to any one of [A1] to [A19], wherein the Caco-2 Papp (cm / sec) value of the compound is equal to or less than the Papp (cm / sec) value of Compound 2. [A21] The preparation or agent according to any one of [A1] to [A20], wherein the Caco-2 Papp (cm / sec) value of the compound is equal to or greater than the Papp (cm / sec) value of Compound 9 and equal to or less than the Papp (cm / sec) value of Compound 2. [A22] The preparation or agent according to any one of [A11] to [A21], wherein the Caco-2 Papp (cm / sec) value is calculated by membrane permeability evaluation under the following conditions: (a) After culturing Caco-2 cells on a 96-well transwell for 3 weeks, the compound to be evaluated, DMEM medium, and FaSSIF (1% DMSO) were added to the apical side, and DMEM medium was added to the basal side, and the cells were incubated under 5% CO. ２(b) After the preincubation, the preincubation solution on the apical and basal sides was removed by suction and washed, and a FaSSIF / HBSS buffer solution (1% DMSO) (pH 6.0) containing a compound was added to the apical side, and an HBSS buffer solution (4% BSA) (pH 7.4) was added to the basal side. (c) Each well was incubated at 37°C under 5% CO ２(d) calculating the permeability coefficient (Caco-2 Papp (cm / sec)) from the measured permeability. [A23] The formulation or agent according to any one of [A1] to [A22], wherein the compound has a solubility of 10 mg / mL or less in 50 mM phosphate buffer (pH 6.5) at 25°C. [A24] The formulation or agent according to any one of [A1] to [A23], wherein the compound has a solubility of 5 mg / mL or less in 50 mM phosphate buffer (pH 6.5) at 25°C. [A25] The formulation or agent according to any one of [A1] to [A24], wherein the compound has a solubility of 2.5 mg / mL or less in 50 mM phosphate buffer (pH 6.5) at 25°C. [A26] The formulation or agent according to any one of [A1] to [A25], wherein the solubility of the compound in 50 mM phosphate buffer (pH 6.5) at 25°C is 2 mg / mL or less. [A27] The formulation or agent according to any one of [A1] to [A26], wherein the molecular weight of the compound is 5,000 or less. [A28] The formulation or agent according to any one of [A1] to [A27], wherein the molecular weight of the compound is 2,000 or less. [A29] The formulation or agent according to any one of [A1] to [A28], wherein the molecular weight of the compound is 500 or more. [A30] The formulation or agent according to any one of [A1] to [A29], wherein the molecular weight of the compound is 1,000 or more. [A31] The formulation or agent according to any one of [A1] to [A30], wherein the molecular weight of the compound is 1,100 or more. [A32] The formulation or agent according to any one of [A1] to [A31], wherein the molecular weight of the compound is 1,200 or more. [A33] The formulation or agent according to any one of [A1] to [A32], wherein the compound has the following characteristics (i) and (ii): (i) the ClogP of the compound is 9 or more, and (ii) the Caco-2 Papp (cm / sec) value of the compound is 1.0E-5 or less.[A34] The formulation or agent according to any one of [A1] to [A32], wherein the compound has the following characteristics (i) and (ii): (i) the ClogP of the compound is 10 or more, (ii) the Caco-2 Papp (cm / sec) value of the compound is 1.0E-7 or more and 1.0E-5 or less. [A35] The formulation or agent according to any one of [A1] to [A32], wherein the compound has the following characteristics (i), (ii), and (iii): (i) the ClogP of the compound is 13 or more, (ii) the Caco-2 Papp (cm / sec) value of the compound is 1.4E-7 or more and 6.0E-6 or less, (iii) the molecular weight of the compound is 1000 or more. [A36] The formulation or agent according to any one of [A1] to [A35], wherein the compound is a peptide compound. [A37] The formulation or agent according to any one of [A1] to [A36], wherein the compound is a peptide compound containing an unnatural amino acid residue. [A38] The formulation or agent according to any one of [A1] to [A37], wherein the compound is a peptide compound containing 5 to 30 amino acid residues. [A39] The formulation or agent according to any one of [A1] to [A38], wherein the compound is a peptide compound containing 7 to 25 amino acid residues. [A40] The formulation or agent according to any one of [A1] to [A39], wherein the compound is a peptide compound containing 8 to 15 amino acid residues. [A41] The formulation or agent according to any one of [A1] to [A40], wherein the compound is a peptide compound containing 9 to 13 amino acid residues. [A42] The formulation or agent according to any one of [A1] to [A41], wherein the compound is a peptide compound containing 11 amino acid residues. [A43] The formulation or agent according to any one of [A36] to [A42], wherein the peptide compound contains two or more N-substituted amino acid residues. [A44] The formulation or agent according to any one of [A36] to [A43], wherein the peptide compound comprises three or more N-substituted amino acid residues. [A45] The formulation or agent according to any one of [A36] to [A44], wherein the peptide compound comprises four or more N-substituted amino acid residues.[A46] The formulation or agent according to any one of [A36] to [A45], wherein the peptide compound contains five or more N-substituted amino acid residues. [A47] The N-substituted amino acid residues are N-C. １ -C ６The formulation or agent according to any one of [A43] to [A46], wherein the N-substituted amino acid residue is an alkyl amino acid residue. [A48] The formulation or agent according to any one of [A43] to [A47], wherein the N-substituted amino acid residue is an N-methyl amino acid residue or an N-ethyl amino acid residue. [A49] The formulation or agent according to any one of [A43] to [A48], wherein the N-substituted amino acid residue is an N-methyl amino acid residue. [A50] The formulation or agent according to any one of [A36] to [A49], wherein the peptide compound is a peptide compound having a cyclic portion. [A51] The formulation or agent according to [A50], wherein the number of amino acid residues constituting the cyclic portion of the peptide compound having a cyclic portion is 5 to 15. [A52] The formulation or agent according to [A50] or [A51], wherein the number of amino acid residues constituting the cyclic portion of the peptide compound having a cyclic portion is 6 to 14. [A53] The formulation or agent according to any one of [A50] to [A52], wherein the number of amino acid residues constituting the cyclic portion of the peptide compound having a cyclic portion is 7 to 14. [A54] The formulation or agent according to any one of [A50] to [A53], wherein the number of amino acid residues constituting the cyclic portion of the peptide compound having a cyclic portion is 8 to 12. [A55] The formulation or agent according to any one of [A50] to [A54], wherein the number of amino acid residues constituting the cyclic portion of the peptide compound having a cyclic portion is 9 to 11. [A56] The formulation or agent according to any one of [A50] to [A55], wherein the number of amino acid residues constituting the cyclic portion of the peptide compound having a cyclic portion is 11. [A57] The formulation or agent according to any one of [A50] to [A56], wherein the number of ring atoms of the peptide compound having a cyclic portion is 34 to 46. [A58] The formulation or agent according to any one of [A50] to [A57], wherein the number of ring atoms of the peptide compound having a cyclic portion is 34 to 43. [A59] The formulation or agent according to any one of [A50] to [A58], wherein the number of ring atoms of the peptide compound having a cyclic portion is 34 to 40. [A60] The formulation according to any one of [A50] to [A59], wherein the peptide compound having a cyclic moiety has 34 to 37 ring atoms.or agent. [A61] The formulation or agent according to any one of [A50] to [A60], wherein the number of ring atoms of the peptide compound having a cyclic portion is 34 or more and 36 or less. [A62] The formulation or agent according to any one of [A50] to [A61], wherein the number of ring atoms of the peptide compound having a cyclic portion is 34. [A63] The formulation or agent according to any one of [A1] to [A62], wherein the compound is a peptide compound having a cyclic portion, and the peptide compound having a cyclic portion has the following characteristics (i) and (ii): (i) comprises a cyclic portion consisting of 5 to 12 amino acid residues, and the number of amino acid residues is 9 to 13, and (ii) comprises at least two N-substituted amino acid residues and at least one non-N-substituted amino acid residue. [A64] The formulation or agent according to any one of [A1] to [A62], wherein the compound is a peptide compound having a cyclic portion, and wherein the peptide compound having a cyclic portion has the following characteristics (i) and (ii): (i) comprises a cyclic portion consisting of 9 to 12 amino acid residues, and the number of amino acid residues is 9 to 13, (ii) comprises at least three N-substituted amino acid residues and at least one amino acid residue that is not N-substituted. [A65] The formulation or agent according to any one of [A1] to [A62], wherein the compound is a peptide compound having a cyclic portion, and wherein the peptide compound having a cyclic portion has the following characteristics (i) and (ii): (i) comprises a cyclic portion consisting of 11 amino acid residues, and the number of amino acid residues is 11, (ii) comprises at least four N-substituted amino acid residues and at least one amino acid residue that is not N-substituted. [A66] The formulation or agent according to any one of [A1] to [A62], wherein the compound is a peptide compound having a cyclic portion, and the peptide compound having a cyclic portion has the following characteristics (i), (ii), (iii), and (iv): (i) it comprises a cyclic portion consisting of 5 to 12 amino acid residues and has 9 to 13 amino acid residues in total, (ii) it comprises at least two N-substituted amino acid residues and at least one non-N-substituted amino acid residue, and (iii) it has a ClogP of 9 or more.(iv) the Caco-2 Papp (cm / sec) value is 1.0E-5 or less. [A67] The formulation or agent according to any of [A1] to [A62], wherein the compound is a peptide compound having a cyclic portion, and the peptide compound having a cyclic portion has the following characteristics (i), (ii), (iii), and (iv): (i) it comprises a cyclic portion consisting of 9 to 12 amino acid residues, and the number of amino acid residues is 9 to 13, (ii) it comprises at least three N-substituted amino acid residues and at least one non-N-substituted amino acid residue, (iii) it has a ClogP of 9 or more, and (iv) it has a Caco-2 Papp (cm / sec) value of 1.0E-5 or less. [A68] The formulation or agent according to any one of [A1] to [A62], wherein the compound is a peptide compound having a cyclic portion, and the peptide compound having a cyclic portion has the following characteristics (i), (ii), (iii), and (iv): (i) it comprises a cyclic portion consisting of 11 amino acid residues, and the number of amino acid residues is 11; (ii) it comprises at least four N-substituted amino acid residues and at least one non-N-substituted amino acid residue; (iii) it has a ClogP of 11 or more; and (iv) it has a Caco-2 Papp (cm / sec) value of 6.0E-6 or less. [A69] The formulation or agent according to any one of [A1] to [A62], wherein the compound is a peptide compound having a cyclic portion, and wherein the peptide compound having a cyclic portion has the following characteristics (i) and (ii): (i) the number of amino acid residues is 9 to 13 and the number of ring atoms in the cyclic portion is 34, (ii) the peptide compound has at least two N-substituted amino acid residues and at least one non-N-substituted amino acid residue. [A70] The formulation or agent according to any one of [A1] to [A62], wherein the compound is a peptide compound having a cyclic portion, and wherein the peptide compound having a cyclic portion has the following characteristics (i), (ii), (iii), (iv) and (v): (i) the number of amino acid residues is 9 to 13 and the number of ring atoms in the cyclic portion is 34, (ii) the peptide compound has at least two N-substituted amino acid residues and at least one non-N-substituted amino acid residue, (iii) the ClogP is 9 or greater.(iv) the Caco-2 Papp (cm / sec) value is 1.0E-5 or less, (v) the molecular weight is 1000 or more. [A71] The formulation or agent according to any of [A1] to [A62], wherein the compound is a peptide compound having a cyclic portion, and the peptide compound having a cyclic portion has the following characteristics (i), (ii), (iii), (iv) and (v): (i) the number of amino acid residues is 11 and the number of ring atoms in the cyclic portion is 34, (ii) the compound contains at least four N-substituted amino acid residues and at least one non-N-substituted amino acid residue, (iii) the ClogP is 13 or more, (iv) the Caco-2 Papp (cm / sec) value is 1.4E-7 or more and 6.0E-6 or less, (v) the molecular weight is 1000 or more. [A72] The formulation or agent according to any one of [A36] to [A71], wherein the peptide compound has a ClogP / amino acid residue ratio of 1.0 or more. [A73] The formulation or agent according to any one of [A36] to [A72], wherein the peptide compound has a ClogP / amino acid residue ratio of 1.0 or more and 1.8 or less. [A74] The peptide compound has a ClogP / amino acid residue ratio of 1.1 or more and 1.6 or less. [A75] The formulation or agent according to any one of [A1] to [A74], wherein the peptide compound is a liquid formulation. [A76] The formulation or agent according to any one of [A1] to [A75], further comprising dimethyl sulfoxide (DMSO). [A77] The formulation or agent according to [A76], wherein the content of DMSO in 100% by volume of the formulation is 1% by volume or more and 30% by volume or less. [A78] The preparation or agent according to [A76] or [A77], wherein the content of the DMSO in 100% by volume of the preparation is 5% by volume or more and 20% by volume or less. [A79] The preparation or agent according to any of [A76] to [A78], wherein the content of the DMSO in 100% by volume of the preparation is 10% by volume. [A80] The preparation according to any of [A1] to [A79], wherein the oily component containing a polyoxyethylene structure is at least one selected from the group consisting of polyoxyethylene castor oil, polyoxyethylene hydrogenated castor oil, and polyoxyethylene sorbitan fatty acid ester.or agent. [A81] A formulation or agent according to any one of [A1] to [A80], wherein the oily component containing a polyoxyethylene structure is polyoxyethylene castor oil. [A82] A formulation or agent according to any one of [A1] to [A81], wherein the oily component containing a polyoxyethylene structure has an average number of added moles of ethylene oxide of 10 or more and 90 or less. [A83] A formulation or agent according to any one of [A1] to [A82], wherein the oily component containing a polyoxyethylene structure has an average number of added moles of ethylene oxide of 15 or more and 60 or less. [A84] A formulation or agent according to any one of [A1] to [A83], wherein the oily component containing a polyoxyethylene structure has an average number of added moles of ethylene oxide of 20 or more and 50 or less. [A85] A formulation or agent according to any one of [A1] to [A84], wherein the oily component containing a polyoxyethylene structure has an average number of added moles of ethylene oxide of 25 or more and 40 or less. [A86] The preparation or agent according to any one of [A1] to [A85], wherein the average number of moles of ethylene oxide added in the oily component containing a polyoxyethylene structure is 30 or more and 39 or less. [A87] The preparation or agent according to any one of [A1] to [A86], wherein the average number of moles of ethylene oxide added in the oily component containing a polyoxyethylene structure is 35. [A88] The preparation or agent according to any one of [A1] to [A87], wherein the components contained in the preparation consist essentially of water, a compound, an oily component containing a polyoxyethylene structure, and DMSO. [A89] The preparation or agent according to any one of [A1] to [A88], wherein the preparation does not contain any surfactant other than the oily component containing a polyoxyethylene structure. [A90] The preparation or agent according to any one of [A1] to [A89], wherein the preparation does not contain lauroyl-L-carnitine. [A91] The preparation or agent according to any one of [A1] to [A90], wherein the compound does not contain cyclosporine or its analogues. [A92] The formulation according to any one of [A1] to [A91], wherein the compound does not contain dihydrocyclosporin D.[A93] The compound is a compound selected from the group consisting of (5S,8S,11S,15S,18S,23aS,29S,35S,37aS)-8-((S)-sec-butyl)-18-cyclopentyl-29-(3,5-difluoro-4-(trifluoromethyl)phenethyl)-36-ethyl-11-isobutyl-N,N,5,6,12,16,19,33-octamethyl-35-(4-methylbenzyl)-4,7,10,13,17,20,23,28,31,34,37-undecaoxotetratriacontahydro-2H,4H-spiro[azeto[2, 1-u]pyrrolo[2,1-i][1,4,7,10,13,16,19,22,25,28,31]undecaazacyclotetratriacontin-21,1'-cyclopentane]-15-carboxamide, (5S,8S,11S,15S,18S,23aS,25R,29S,35S,37aS)-8-((S)-sec-butyl)-35-(cyclohexylmethyl)-18-cyclopentyl-29-(3,5-difluoro-4-(trifluoromethyl)phenethyl)-25-ethoxy-11-isobutyl-N,N , 5,6,12,16,19,33,36-nonamethyl-4,7,10,13,17,20,23,28,31,34,37-undecaoxotetratriacontahydro-2H,4H-spiro[azeto[2,1-u]pyrrolo[2,1-i][1,4,7,10,13,16,19,22,25,28,31]undecaazacyclotetratriacontin-21,1'-cyclopentane]-15-carboxamide, (3S,9S,18S,21S,25S,28S,34S)-3-[2-[3-chloro-4-(trifluoromethyl)methyl]phenyl]propan-1-yl]propan-2-yl]propan-1 ... (fluoromethyl)phenyl]ethyl]-28-cyclohexyl-9-(cyclohexylmethyl)-21-isobutyl-7,10,13,16,22,26,29-heptamethyl-18-[(1S)-1-methylpropyl]-25-(piperidine-1-carbonyl)spiro[1,4,7,10,13,16,19,22,26,29,32-undecazabicyclo[32.3.0]heptatriacontane-31,1′-cyclopentane]-2,5,8,11,14,17,20,23,27,30,33-undecaone,(5S,8S,11S,15S,18S,23aS,29S,35S,37aS)-8-((S)-sec-butyl)-29-(3-chloro-4-(trifluoromethyl)phenethyl)-35-(cyclohexylmethyl)-18-cyclopentyl-11-isobutyl-5,6,12,16,19,33,36-heptamethyl-15-(morpholine-4-carbonyl)doco sahydro-2H,4H-spiro[azeto[2,1-u]pyrrolo[2,1-i][1,4,7,10,13,16,19,22,25,28,31]undecaazacyclotetratriacontin-21,1′-cyclopentane]-4,7,10,13,17,20,23,28,31,34,37(14H,22H)-undecaone, and (5S,8S,11S,15S,1 8S,23aS,29S,35S,37aS)-8-((S)-sec-butyl)-29-(3-chloro-4-(trifluoromethyl)phenethyl)-18-cyclopentyl-36-ethyl-11-isobutyl-5,6,12,16,19,33-hexamethyl-35-(4-methylbenzyl)-15-(morpholine-4-carbonyl)docosahydro-2H,4H-spiro[4-methylbenzyl]methyl] The formulation or agent according to any one of [A1] to [A92], which does not contain 2[azeto[2,1-u]pyrrolo[2,1-i][1,4,7,10,13,16,19,22,25,28,31]undecaazacyclotetratriacontin-21,1'-cyclopentane]-4,7,10,13,17,20,23,28,31,34,37(14H,22H)-undecaone. [A94] The formulation or agent according to any one of [A1] to [A93], wherein the formulation exhibits enhanced absorption of the compound compared to a formulation not containing an oily component containing a polyoxyethylene structure. [A95] The formulation or agent according to any one of [A1] to [A94], wherein the formulation is an oral formulation. [A96] The formulation or agent according to any one of [A1] to [A95], wherein the formulation is used for screening candidate compounds. [A97] The preparation or agent according to any one of [A1] to [A96], wherein the content of the oily component in 100% by volume of the preparation is 2.0% by volume or more and less than 7.5% by volume. [A98] The content of the oily component in 100% by volume of the preparation is 2.0% by volume or more and 7.0% by volume or less.The preparation or agent according to any one of [A1] to [A96]. [A99] The preparation or agent according to any one of [A1] to [A96], wherein the content of the oily component in 100% by volume of the preparation is 2.0% by volume or more and 5.0% by volume or less.

[0009] One aspect of the present invention relates to a method for screening a candidate compound. Specifically, for example, the present invention encompasses the following inventions. [B1] A method for screening a candidate compound, comprising administering the formulation according to any one of [A1] to [A99] to a subject. [B2] The method according to [B1], comprising selecting the candidate compound using at least one index selected from the group consisting of (1), (2), and (3) below: (1) efficacy, (2) toxicity, (3) pharmacokinetic properties. [B3] The method according to [B2], wherein the index is (3) pharmacokinetic properties. [B4] The method according to any one of [B1] to [B3], wherein the candidate compound is selected from a plurality of test compounds. [B5] The method according to any one of [B1] to [B4], wherein the candidate compound is selected from three or more test compounds. [B6] The method according to any one of [B1] to [B5], wherein the candidate compound is a pharmaceutical candidate compound. [B7] The method according to any one of [B1] to [B6], wherein the subject is a non-human animal. [B8] The method according to any one of [B1] to [B7], wherein the subject is at least one selected from the group consisting of dogs, monkeys, minipigs, rabbits, rats, and mice. [B9] The method according to any one of [B1] to [B8], wherein the subject is at least one selected from the group consisting of rats and mice. [B10] The method according to any one of [B1] to [B9], wherein the subject is a mouse. [B11] The method according to any one of [B1] to [B10], wherein the administration is oral administration. [B12] The method according to any one of [B1] to [B11], wherein the administered dose of the test compound is 0.1 mg / kg or more and 1000 mg / kg or less. [B13] The method according to any one of [B1] to [B11], wherein the administered dose of the test compound is 1 mg / kg or more and 500 mg / kg or less. [B14] The method according to any one of [B1] to [B11], wherein the administered dose of the test compound is 1 mg / kg or more and 100 mg / kg or less. [B15] The method according to any one of [B1] to [B11], wherein the test compound is administered at a dose of 1 mg / kg or more and 50 mg / kg or less.[B16] The method according to any one of [B1] to [B11], wherein the test compound is administered at a dose of 3 mg / kg or more and 30 mg / kg or less. [B17] The method according to any one of [B1] to [B11], wherein the test compound is administered at a dose of 10 mg / kg or more and 30 mg / kg or less. [B18] The method according to any one of [B1] to [B11], wherein the test compound is administered at a dose of 0.1 mg / kg or more and 10 mg / kg or less. [B19] The method according to any one of [B1] to [B11], wherein the test compound is administered at a dose of 1 mg / kg or more and 5 mg / kg or less. [B20] The method according to any one of [B1] to [B11], wherein the test compound is administered at a dose of 10 mg / kg or more and 100 mg / kg or less. [B21] The method according to any one of [B1] to [B11], wherein the test compound is administered at a dose of 15 mg / kg or more and 50 mg / kg or less. [B22] The method according to any one of [B1] to [B11], wherein the test compound is administered at a dose of 20 mg / kg or more and 40 mg / kg or less. [B23] The method according to any one of [B1] to [B11], wherein the test compound is administered at a dose of 25 mg / kg or more and 35 mg / kg or less. [B24] The method according to any one of [B1] to [B11], wherein the test compound is administered at a dose of 3 mg / kg. [B25] The method according to any one of [B1] to [B11], wherein the test compound is administered at a dose of 30 mg / kg.

[0010] One aspect of the present invention relates to a method for comparing test compounds. Specifically, the present invention encompasses the following inventions: [C1] A method for comparing multiple test compounds, comprising administering to the subject a formulation according to any one of [A1] to [A99], and comparing the multiple test compounds using at least one index selected from the group consisting of (1), (2), and (3) below: (1) efficacy, (2) toxicity, or (3) pharmacokinetic properties, wherein the formulation or agent includes one test compound. [C2] The method according to [C1], wherein the index is (3) pharmacokinetic properties. [C3] The method according to [C1] or [C2], wherein the subject is a non-human animal. [C4] The method according to any one of [C1] to [C3], wherein the subject is at least one selected from the group consisting of dogs, monkeys, minipigs, rabbits, rats, and mice. [C5] The method according to any one of [C1] to [C4], wherein the subject is at least one selected from the group consisting of rats and mice. [C6] The method according to any one of [C1] to [C5], wherein the subject is a mouse. [C7] The method according to any one of [C1] to [C6], wherein the administration is oral. [C8] The method according to any one of [C1] to [C7], wherein the test compound is administered at a dose of 0.1 mg / kg to 1000 mg / kg. [C9] The method according to any one of [C1] to [C7], wherein the test compound is administered at a dose of 1 mg / kg to 500 mg / kg. [C10] The method according to any one of [C1] to [C7], wherein the test compound is administered at a dose of 1 mg / kg to 100 mg / kg. [C11] The method according to any one of [C1] to [C7], wherein the test compound is administered at a dose of 1 mg / kg to 50 mg / kg. [C12] The method according to any one of [C1] to [C7], wherein the test compound is administered at a dose of 3 mg / kg to 30 mg / kg. [C13] The method according to any one of [C1] to [C7], wherein the test compound is administered at a dose of 10 mg / kg or more and 30 mg / kg or less. [C14] The method according to any one of [C1] to [C7], wherein the test compound is administered at a dose of 0.1 mg / kg or more and 10 mg / kg or less.[C15] The method according to any one of [C1] to [C7], wherein the test compound is administered at a dose of 1 mg / kg or more and 5 mg / kg or less. [C16] The method according to any one of [C1] to [C7], wherein the test compound is administered at a dose of 10 mg / kg or more and 100 mg / kg or less. [C17] The method according to any one of [C1] to [C7], wherein the test compound is administered at a dose of 15 mg / kg or more and 50 mg / kg or less. [C18] The method according to any one of [C1] to [C7], wherein the test compound is administered at a dose of 20 mg / kg or more and 40 mg / kg or less. [C19] The method according to any one of [C1] to [C7], wherein the test compound is administered at a dose of 25 mg / kg or more and 35 mg / kg or less. [C20] The method according to any one of [C1] to [C7], wherein the test compound is administered at a dose of 3 mg / kg or less. [C21] The method according to any one of [C1] to [C7], wherein the test compound is administered at a dose of 30 mg / kg.

[0011] One aspect of the present invention relates to a method for enhancing systemic absorption of a test compound. Specifically, for example, the present invention encompasses the following inventions. [D1] A method for enhancing systemic absorption of a test compound in a subject, comprising administering to the subject a formulation according to any one of [A1] to [A96]. [D2] The method according to [D1], wherein the systemic absorption is gastrointestinal absorption. [D3] The method according to [D1] or [D2], wherein the subject is a non-human animal. [D4] The method according to any one of [D1] to [D3], wherein the subject is at least one selected from the group consisting of dogs, monkeys, minipigs, rabbits, rats, and mice. [D5] The method according to any one of [D1] to [D4], wherein the subject is at least one selected from the group consisting of rats and mice. [D6] The method according to any one of [D1] to [D5], wherein the subject is a mouse. [D7] The method according to any one of [D1] to [D6], wherein the administration is oral administration. [D8] The method according to any one of [D1] to [D7], wherein the test compound is administered at a dose of 0.1 mg / kg or more and 1000 mg / kg or less. [D9] The method according to any one of [D1] to [D7], wherein the test compound is administered at a dose of 1 mg / kg or more and 500 mg / kg or less. [D10] The method according to any one of [D1] to [D7], wherein the test compound is administered at a dose of 1 mg / kg or more and 100 mg / kg or less. [D11] The method according to any one of [D1] to [D7], wherein the test compound is administered at a dose of 1 mg / kg or more and 50 mg / kg or less. [D12] The method according to any one of [D1] to [D7], wherein the test compound is administered at a dose of 3 mg / kg or more and 30 mg / kg or less. [D13] The method according to any one of [D1] to [D7], wherein the test compound is administered at a dose of 10 mg / kg or more and 30 mg / kg or less. [D14] The method according to any one of [D1] to [D7], wherein the test compound is administered at a dose of 0.1 mg / kg or more and 10 mg / kg or less. [D15] The method according to any one of [D1] to [D7], wherein the test compound is administered at a dose of 1 mg / kg or more and 5 mg / kg or less.[D16] The method according to any one of [D1] to [D7], wherein the test compound is administered at a dose of 10 mg / kg or more and 100 mg / kg or less. [D17] The method according to any one of [D1] to [D7], wherein the test compound is administered at a dose of 15 mg / kg or more and 50 mg / kg or less. [D18] The method according to any one of [D1] to [D7], wherein the test compound is administered at a dose of 20 mg / kg or more and 40 mg / kg or less. [D19] The method according to any one of [D1] to [D7], wherein the test compound is administered at a dose of 25 mg / kg or more and 35 mg / kg or less. [D20] The method according to any one of [D1] to [D7], wherein the test compound is administered at a dose of 3 mg / kg. [D21] The method according to any one of [D1] to [D7], wherein the test compound is administered at a dose of 30 mg / kg.

[0012] One aspect of the present invention relates to a method for producing a formulation used in screening for a candidate compound. Specifically, for example, the present invention encompasses the following inventions. [E1] A method for producing a formulation used in screening for a candidate compound, comprising blending a test compound and an oily component containing a polyoxyethylene structure, wherein the oily component is blended so that its content in 100% by volume of the formulation is 2.0% by volume or more and 7.5% by volume or less, the test compound is a compound described in any of [A1] to [A96], and the formulation is a formulation described in any of [A1] to [A96]. [E2] The method for producing the formulation described in [E1], wherein the components contained in the formulation consist essentially of water, the test compound, the oily component containing a polyoxyethylene structure, and DMSO. [E3] The method for producing the formulation described in [E1] or [E2], wherein the formulation is a liquid formulation. [E4] The method for producing the formulation described in any of [E1] to [E3], wherein the formulation is an oral formulation.

[0013] According to the present invention, by applying a formulation containing an oily component containing a polyoxyethylene structure in a range of 2.0% to 7.5% by volume based on 100% by volume of the formulation, the oral absorbability of a compound can be generally improved. This makes it possible to evaluate different compounds under the same formulation conditions. Furthermore, by using this formulation, candidate compounds can be screened while reducing the risk of overlooking useful compounds as false negatives.

[0014] 1 is a graph showing the relationship between polyoxyethylene castor oil (average number of moles of ethylene oxide added: 35) concentration and AUC ratio (30 mg / kg). The dashed lines in the graph represent concentrations of 2.0 vol% and 7.5 vol% polyoxyethylene castor oil (average number of moles of ethylene oxide added: 35). FIG. 1 is a graph showing the relationship between polyoxyethylene castor oil (average number of moles of ethylene oxide added: 35) concentration and AUC ratio (3 mg / kg). The dashed lines in the graph represent concentrations of 2.0 vol% and 7.5 vol% polyoxyethylene castor oil (average number of moles of ethylene oxide added: 35).

[0015] Hereinafter, embodiments of the present invention will be described in detail, but the present invention is not limited to the following embodiments.

[0016] As used herein, "one or more" means one or more than one. When "one or more" is used in the context of substituents on a group, the term means a number from one to the maximum number of substituents permitted by that group. Specific examples of "one or more" include 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, and / or more.

[0017] In this specification, the term "to" indicating a range includes both ends of the range. For example, "A to B" means a range equal to or greater than A and equal to or less than B.

[0018] As used herein, the term "about" when used in conjunction with a numerical value means a range of values ​​of plus and minus 10% of that numerical value.

[0019] In the present invention, the meaning of the term "and / or" includes any combination of "and" and "or" appropriately combined. Specifically, for example, "A, B and / or C" includes the following seven variations: (i) A, (ii) B, (iii) C, (iv) A and B, (v) A and C, (vi) B and C, and (vii) A, B and C.

[0020] [Formulation] The formulation according to this embodiment contains a compound and an oily component containing a polyoxyethylene structure, and the content of the oily component containing a polyoxyethylene structure in 100% by volume of the formulation is 2.0% by volume or more and 7.5% by volume or less.

[0021] (Compound) The compound contained in the formulation according to this embodiment is not particularly limited, and any compound can be used. Meanwhile, since the formulation according to this embodiment can generally increase the oral absorbability of compounds, compounds with low solubility and / or membrane permeability can be suitably used as the compound contained in the formulation according to this embodiment.

[0022] The compound of one embodiment may have a ClogP of 4 or more and 25 or less. ClogP is a computer-calculated partition coefficient, and can be determined in accordance with the principles described in "CLOGP Reference Manual Daylight Version 4.9 (Release Date: August 1, 2011, https: / / www.daylight.com / dayhtml / doc / clogp / )." As an example of a method for calculating ClogP, the ClogP is calculated using the CLOGP Reference Manual published by Daylight Chemical Information Systems, Inc. Examples of calculations include using Daylight Version 4.95 (release date: August 1, 2011, ClogP algorithm version 5.4, database version 28, https: / / www.daylight.com / dayhtml / doc / release_notes / index.html / ) by Yahoo!

[0023] The ClogP of the compound according to this embodiment may be, for example, 24 or less, 23 or less, 22 or less, 21 or less, 20 or less, 19 or less, 18 or less, 17 or less, or 16.1 or less. The lower limit of the ClogP of the compound according to this embodiment may be, for example, 5 or more, 6 or more, 7 or more, 8 or more, 9 or more, 10 or more, 11 or more, 12 or more, 13 or more, or 13.2 or more. Examples of the range of the ClogP of the compound according to this embodiment include 5 or more and 24 or less, 6 or more and 23 or less, 7 or more and 22 or less, 8 or more and 21 or less, 9 or more and 20 or less, 10 or more and 20 or less, 11 or more and 19 or less, 12 or more and 18 or less, 13 or more and 17 or less, and 13.2 or more and 16.1 or less. The ClogP of the compound according to this embodiment may also be, for example, greater than the ClogP of Compound 2 and less than the ClogP of Compound 1, or greater than the ClogP of Compound 2 and less than the ClogP of Compound 1.

[0024] The compound of one embodiment may have a Caco-2 Papp (cm / sec) value of 1.0E-9 or greater.

[0025] The Caco-2 Papp (cm / sec) value is an index of membrane permeability in the cell membrane and can be measured by the following method: (a) After culturing Caco-2 cells on a 96-well transwell for 3 weeks, the compound to be evaluated, DMEM medium, and FaSSIF (1% DMSO) were added to the apical side, and DMEM medium was added to the basal side. ２ (b) After preincubation, the preincubation solution on the apical and basal sides is removed by suction and washed. A FaSSIF / HBSS buffer solution (1% DMSO) (pH 6.0) containing the compound is added to the apical side, and HBSS buffer solution (4% BSA) (pH 7.4) is added to the basal side. (c) Each well is maintained at 37°C with 5% CO ２The tube is shaken at 80 rpm at 37°C under atmospheric pressure, and a sample is taken from the basal side 180 minutes after the start of shaking, and the amount of compound permeated is measured by liquid chromatography mass spectrometry (LC / MS). (d) The permeability coefficient (Caco-2 Papp (cm / sec)) is calculated from the measured permeability. The permeability coefficient (Caco-2 Papp (cm / sec)) is calculated using Equation 1. In the above measurement, a Pgp inhibitor (e.g., Zosquidar) can be added to FaSSIF / HBSS buffer (1% DMSO) (pH 6.0) and HBSS buffer (4% BSA) (pH 7.4), respectively. The concentration of the donor-side target compound used to calculate the permeability coefficient in step (d) above can be the concentration of the compound initially added, or the compound concentration measured by sampling the apical solution before starting preincubation or before starting shaking in step (c) above can also be used. In particular, when preincubation is performed, it is preferable to use the concentration of the compound sampled from the apical side before preincubation.

[0026] The Caco-2 Papp (cm / sec) value of the compound according to this embodiment may be, for example, 1.0E-4 or less, 9.0E-5 or less, 8.0E-5 or less, 7.0E-5 or less, 6.0E-5 or less, 5.0E-5 or less, 4.0E-5 or less, 3.0E-5 or less, 2.0E-5 or less, 1.0E-5 or less, 9.0E-6 or less, 8.0E-6 or less, 7.0E-6 or less, 6.0E-6 or less, or 5.99E-6 or less. The lower limit of the Caco-2 Papp (cm / sec) value of the compound according to this embodiment may be, for example, 1.0E-9 or more, 1.0E-8 or more, 2.0E-8 or more, 3.0E-8 or more, 4.0E-8 or more, 5.0E-8 or more, 6.0E-8 or more, 7.0E-8 or more, 8.0E-8 or more, 9.0E-8 or more, 1.0E-7 or more, 1.1E-7 or more, 1.2E-7 or more, 1.3E-7 or more, or 1.4E-7 or more. Examples of the range of the Caco-2 Papp (cm / sec) value of the compound according to this embodiment include 1.0E-9 or more and 1.0E-4 or less, 1.0E-8 or more and 1.0E-5 or less, 1.0E-7 or more and 1.0E-5 or less, and 1.4E-7 or more and 5.99E-6 or less. Note that E-n (n is a natural number) is 10 －ｎ (For example, 1.0E-5 = 1.0 × 10 －５ The Caco-2 Papp (cm / sec) value of the compound according to this embodiment may also be, for example, equal to or greater than the Papp (cm / sec) value of Compound 9, and equal to or less than the Papp (cm / sec) value of Compound 2, or equal to or greater than the Papp (cm / sec) value of Compound 9 but equal to or less than the Papp (cm / sec) value of Compound 2.

[0027] The compound of one embodiment may have a solubility of 10 mg / mL or less in 50 mM phosphate buffer (pH 6.5) at 25° C. The solubility of the compound of this embodiment in 50 mM phosphate buffer (pH 6.5) at 25° C. may be, for example, 5 mg / mL or less, 2.5 mg / mL or less, 2 mg / mL or less, 1 mg / mL or less, 0.5 mg / mL or less, 0.25 mg / mL or less, 0.1 mg / mL or less, 0.05 mg / mL or less, 0.025 mg / mL or less, 0.01 mg / mL or less, 0.005 mg / mL or less, 0.0025 mg / mL or less, or 0.001 mg / mL or less.

[0028] The compound of one embodiment may be a compound classified as Class IV in the Biopharmaceutics Classification System, which is a guideline for predicting the gastrointestinal absorption characteristics of pharmaceuticals by classifying them into four classes (Class I to Class IV) based on their solubility and absorption rate.

[0029] Solubility in BCS (D ０ : Dose Number) is D ０ = 1 as the boundary, D ０ When D is less than 1, the solubility is judged to be high (high solubility), and ０ When the value is ≧1, the solubility is judged to be low (low solubility). ａ F: the rate of absorption from the digestive tract) is measured with a 90% absorption rate as the boundary. ａ When the absorption rate is ≦0.9, it is determined that the absorption rate is low (low absorption rate), and F ａ When the D solubility is ≥ 0.9, the absorption rate is judged to be high (high absorption rate). BCS Classes I to IV are defined as follows: Class I: High solubility (D ０ ≦1) and high absorption rate (F ａ ≧0.9) Class II: Low solubility (D ０ ≧1) and high absorption rate (F ａ ≧0.9) Class III: High solubility (D ０ ≦1) and low absorption rate (F ａ ≦0.9) Class IV: Low solubility (D ０ ≧1) and low absorption rate (F ａ ≦0.9)

[0030] The compound according to one embodiment may have a molecular weight of 5,000 or less. The molecular weight in this specification refers to the sum of the atomic weights of the atoms constituting the compound molecule (unit: g / mol), and is obtained by calculating the sum of the atomic weights of the atoms included in the molecular structure (unit: g / mol). Hereinafter, in this specification, the unit of molecular weight may be omitted.

[0031] The molecular weight of the compound according to this embodiment is not particularly limited, but may be, for example, 500 or more, 550 or more, 600 or more, 650 or more, 700 or more, 750 or more, 800 or more, 850 or more, 900 or more, 950 or more, 1000 or more, 1100 or more, 1200 or more, 1300 or more, or 1400 or more. The upper limit of the molecular weight of the compound according to this embodiment is not particularly limited, but may be 5000 or less, 4000 or less, 3000 or less, 2500 or less, 2000 or less, 1900 or less, 1800 or less, 1700 or less, or 1600 or less. Examples of molecular weight ranges for the compound according to this embodiment include 500 to 5000, 750 to 4000, 1000 to 3000, 1100 to 2000, 1200 to 1800, and 1400 to 1600.

[0032] A compound according to one embodiment may be a peptide compound. As used herein, the term "peptide compound" is not particularly limited as long as the amino acid residues are linked by amide bonds or ester bonds, but preferably are linked by amide bonds. The number of amino acid residues in a peptide compound is not particularly limited, but is preferably 5 or more, more preferably 7 or more, even more preferably 8 or more, and even more preferably 9 or more. The number of amino acid residues in a peptide compound is also preferably 30 or less, more preferably 25 or less, even more preferably 15 or less, and even more preferably 13 or less. The number of amino acid residues in a peptide compound may be, for example, 5 to 30, 7 to 25, 8 to 15, 9 to 13, or 11. The peptide compound may have a branched structure.

[0033] In this specification, the "amino acid residues" that constitute a peptide compound may be simply referred to as "amino acids".

[0034] As used herein, "amino acid" includes natural amino acids and unnatural amino acids. Furthermore, as used herein, "amino acid" may refer to an amino acid residue. Furthermore, as used herein, "amino acid residue" includes natural amino acid residues and unnatural amino acid residues.

[0035] Naturally occurring amino acids refer to glycine (Gly), alanine (Ala), serine (Ser), threonine (Thr), valine (Val), leucine (Leu), isoleucine (Ile), phenylalanine (Phe), tyrosine (Tyr), tryptophan (Trp), histidine (His), glutamic acid (Glu), aspartic acid (Asp), glutamine (Gln), asparagine (Asn), cysteine ​​(Cys), methionine (Met), lysine (Lys), arginine (Arg), and proline (Pro).

[0036] Examples of unnatural amino acids include, but are not limited to, β-amino acids, D-amino acids, N-substituted amino acids (excluding Pro), α,α-disubstituted amino acids, amino acids whose side chains differ from those of natural amino acids, hydroxycarboxylic acids, etc. As used herein, unnatural N-substituted amino acids refer to N-substituted amino acids other than Pro.

[0037] The amino acids herein may have any steric configuration. The side chain of the amino acid is not particularly limited, and may be freely selected from, in addition to a hydrogen atom, for example, an alkyl group, an alkenyl group, an alkynyl group, an aryl group, a heteroaryl group, an aralkyl group, a heteroaralkyl group, a cycloalkyl group, or a spiro-linked cycloalkyl group. Each of these groups may be substituted, and the substituents are not limited thereto. For example, one or more groups may be independently selected from any substituent containing a halogen atom, an O atom, a S atom, a N atom, a B atom, a Si atom, or a P atom. Examples of such groups include optionally substituted alkyl groups, alkoxy groups, alkoxyalkyl groups, alkenyl groups, alkynyl groups, aryl groups, heteroaryl groups, aralkyl groups, cycloalkyl groups, etc., or oxo, aminocarbonyl, halogen atoms, etc. In a non-limiting embodiment, the amino acids herein may be compounds having a carboxyl group and an amino group in the same molecule (even in this case, imino acids such as proline and hydroxyproline are also included in the amino acid).

[0038] Halogen-derived substituents include fluoro (-F), chloro (-Cl), bromo (-Br), iodo (-I), and the like.

[0039] Substituents derived from O atoms include hydroxy (-OH), oxy (-OR), carbonyl (-C(=O)-R), carboxy (-CO ２ H), oxycarbonyl (-C(=O)-OR), carbonyloxy (-O-C(=O)-R), thiocarbonyl (-C(=O)-SR), carbonylthio group (-S-C(=O)-R), aminocarbonyl (-C(=O)-NHR), carbonylamino (-NH-C(=O)-R), oxycarbonylamino (-NH-C(=O)-OR), sulfonylamino (-NH-SO ２ -R), aminosulfonyl (-SO ２ -NHR), sulfamoylamino (-NH-SO ２ -NHR), thiocarboxy (-C(=O)-SH), carboxylcarbonyl (-C(=O)-CO ２ H).

[0040] Examples of oxy (—OR) include alkoxy, cycloalkoxy, alkenyloxy, alkynyloxy, aryloxy, heteroaryloxy, aralkyloxy, and the like.

[0041] Examples of carbonyl (-C(=O)-R) include formyl (-C(=O)-H), alkylcarbonyl, cycloalkylcarbonyl, alkenylcarbonyl, alkynylcarbonyl, arylcarbonyl, heteroarylcarbonyl, aralkylcarbonyl, and the like.

[0042] Examples of oxycarbonyl (-C(=O)-OR) include alkyloxycarbonyl, cycloalkyloxycarbonyl, alkenyloxycarbonyl, alkynyloxycarbonyl, aryloxycarbonyl, heteroaryloxycarbonyl, aralkyloxycarbonyl, and the like.

[0043] Examples of carbonyloxy (—O—C(═O)—R) include alkylcarbonyloxy, cycloalkylcarbonyloxy, alkenylcarbonyloxy, alkynylcarbonyloxy, arylcarbonyloxy, heteroarylcarbonyloxy, aralkylcarbonyloxy, and the like.

[0044] Examples of thiocarbonyl (-C(=O)-SR) include alkylthiocarbonyl, cycloalkylthiocarbonyl, alkenylthiocarbonyl, alkynylthiocarbonyl, arylthiocarbonyl, heteroarylthiocarbonyl, aralkylthiocarbonyl, and the like.

[0045] Examples of carbonylthio (-S-C(=O)-R) include alkylcarbonylthio, cycloalkylcarbonylthio, alkenylcarbonylthio, alkynylcarbonylthio, arylcarbonylthio, heteroarylcarbonylthio, aralkylcarbonylthio, and the like.

[0046] Examples of aminocarbonyl (-C(=O)-NHR) include alkylaminocarbonyl, cycloalkylaminocarbonyl, alkenylaminocarbonyl, alkynylaminocarbonyl, arylaminocarbonyl, heteroarylaminocarbonyl, aralkylaminocarbonyl, etc. In addition to these, compounds in which the H atom bonded to the N atom in -C(=O)-NHR is further substituted with an alkyl, cycloalkyl, alkenyl, alkynyl, aryl, heteroaryl, or aralkyl are also included.

[0047] Examples of carbonylamino (-NH-C(=O)-R) include alkylcarbonylamino, cycloalkylcarbonylamino, alkenylcarbonylamino, alkynylcarbonylamino, arylcarbonylamino, heteroarylcarbonylamino, aralkylcarbonylamino, etc. In addition to these, compounds in which the H atom bonded to the N atom in -NH-C(=O)-R is further substituted with alkyl, cycloalkyl, alkenyl, alkynyl, aryl, heteroaryl, or aralkyl are also included.

[0048] Examples of oxycarbonylamino (-NH-C(=O)-OR) include alkoxycarbonylamino, cycloalkoxycarbonylamino, alkenyloxycarbonylamino, alkynyloxycarbonylamino, aryloxycarbonylamino, heteroaryloxycarbonylamino, aralkyloxycarbonylamino, etc. In addition to these, examples include compounds in which the H atom bonded to the N atom in -NH-C(=O)-OR is further substituted with an alkyl, cycloalkyl, alkenyl, alkynyl, aryl, heteroaryl, or aralkyl.

[0049] Sulfonylamino (-NH-SO ２ Examples of —R) include alkylsulfonylamino, cycloalkylsulfonylamino, alkenylsulfonylamino, alkynylsulfonylamino, arylsulfonylamino, heteroarylsulfonylamino, aralkylsulfonylamino, etc. In addition to these, —NH—SO ２ Examples include compounds in which the H atom bonded to the N atom in —R is further substituted with alkyl, cycloalkyl, alkenyl, alkynyl, aryl, heteroaryl, or aralkyl.

[0050] Aminosulfonyl (-SO ２ Examples of —NHR) include alkylaminosulfonyl, cycloalkylaminosulfonyl, alkenylaminosulfonyl, alkynylaminosulfonyl, arylaminosulfonyl, heteroarylaminosulfonyl, aralkylaminosulfonyl, and the like. ２ Examples include compounds in which the H atom bonded to the N atom in —NHR is further substituted with alkyl, cycloalkyl, alkenyl, alkynyl, aryl, heteroaryl, or aralkyl.

[0051] Sulfamoylamino (-NH-SO ２Examples of —NHR) include alkylsulfamoylamino, cycloalkylsulfamoylamino, alkenylsulfamoylamino, alkynylsulfamoylamino, arylsulfamoylamino, heteroarylsulfamoylamino, and aralkylsulfamoylamino. ２ The two H atoms bonded to the N atom in —NHR may be substituted with substituents independently selected from the group consisting of alkyl, cycloalkyl, alkenyl, alkynyl, aryl, heteroaryl, and aralkyl, and these two substituents may form a ring.

[0052] Substituents derived from S atoms include thiol (-SH), thio (-S-R), sulfinyl (-S(=O)-R), sulfonyl (-S(O) ２ -R), sulfo (-SO ３ H), pentafluorosulfanyl (-SF ５ ) etc.

[0053] Examples of thio (-S-R) include alkylthio, cycloalkylthio, alkenylthio, alkynylthio, arylthio, heteroarylthio, aralkylthio, and the like.

[0054] Examples of sulfinyl (-S(=O)-R) include alkylsulfinyl, cycloalkylsulfinyl, alkenylsulfinyl, alkynylsulfinyl, arylsulfinyl, heteroarylsulfinyl, aralkylsulfinyl, and the like.

[0055] Sulfonyl (-S(O) ２ Examples of —R) include alkylsulfonyl, cycloalkylsulfonyl, alkenylsulfonyl, alkynylsulfonyl, arylsulfonyl, heteroarylsulfonyl, aralkylsulfonyl, and the like.

[0056] As a substituent derived from the N atom, azide (-N ３ , also called "azido group"), cyano (-CN), primary amino (-NH ２ ), secondary amino (—NH—R), tertiary amino (—NR(R′)), amidino (—C(═NH)—NH２ ), substituted amidino (—C(═NR)—NR′R″), guanidino (—NH—C(═NH)—NH ２ ), substituted guanidino (—NR—C(═NR′″)—NR′R″), aminocarbonylamino (—NR—CO—NR′R″), and the like.

[0057] Examples of secondary amino (-NH-R) include alkylamino, cycloalkylamino, alkenylamino, alkynylamino, arylamino, heteroarylamino, and aralkylamino.

[0058] Examples of tertiary amino (—NR(R′)) include an amino group having any two substituents independently selected from alkyl, cycloalkyl, alkenyl, alkynyl, aryl, heteroaryl, aralkyl, etc., such as alkyl(aralkyl)amino, and these two substituents may form a ring.

[0059] Examples of substituted amidino (-C(=NR)-NR'R'') include groups in which the three substituents R, R', and R'' on the N atom are each independently selected from alkyl, cycloalkyl, alkenyl, alkynyl, aryl, heteroaryl, and aralkyl, such as alkyl(aralkyl)(aryl)amidino.

[0060] Examples of substituted guanidino (-NR-C(=NR'")-NR'R") include groups in which R, R', R", and R'" are each independently selected from alkyl, cycloalkyl, alkenyl, alkynyl, aryl, heteroaryl, and aralkyl, and groups in which these groups form a ring.

[0061] Examples of aminocarbonylamino (—NR—CO—NR′R″) include groups in which R, R′, and R″ are each independently selected from a hydrogen atom, alkyl, cycloalkyl, alkenyl, alkynyl, aryl, heteroaryl, and aralkyl, and groups in which these groups form a ring.

[0062] Examples of the substituent derived from the B atom include boryl (-BR(R')) and dioxyboryl (-B(OR)(OR')). These two substituents R and R' may be groups independently selected from alkyl, cycloalkyl, alkenyl, alkynyl, aryl, heteroaryl, aralkyl, etc., or may be a group in which these groups form a ring. Specific examples include cyclic boryl groups, and more specific examples include pinacolatoboryl groups, neopentanediolateboryl groups, and catecholateboryl groups.

[0063] The main chain amino group of the amino acid is unsubstituted (-NH ２ ) or may be substituted (i.e., —NHR, where R represents, for example, an alkyl group, alkenyl group, alkynyl group, aryl group, heteroaryl group, aralkyl group, cycloalkyl group, or the like, which may have a substituent, and the carbon chain bonded to the N atom and the carbon atom at the α-position may form a ring, as in proline).

[0064] In the present specification, an amino acid in which the main chain amino group is substituted is referred to as an "N-substituted amino acid." As used herein, the "N-substituted amino acid" is preferably an N-alkyl amino acid, an N-C １ -C ６ Alkylamino acid, N-C １ -C ５ Alkylamino acid, N-C １ -C ４ Alkylamino acid, N-C １ -C ３ Alkyl amino acids, N-ethyl amino acids, N-methyl amino acids, N-C ７ -C １４ Examples include, but are not limited to, aralkyl amino acids, N-benzyl amino acids, and N-phenethyl amino acids.

[0065] Specific examples of the substituent on the nitrogen atom of the N-substituted amino acid in this specification (R in the above-mentioned —NHR) include alkyl groups (preferably C １ -C ６ alkyl group, more preferably C １ -C ４ alkyl group, more preferably C １ -C３ an alkyl group, more preferably an ethyl group or a methyl group), C ７ -C １４ Examples include an aralkyl group, a benzyl group, a phenethyl group, etc. As the substituent on the nitrogen atom of the N-substituted amino acid, an ethyl group or a methyl group is more preferred, and a methyl group is particularly preferred (i.e., an N-methyl amino acid is particularly preferred as the N-substituted amino acid).

[0066] As used herein, "amino acids" include all corresponding isotopes. An isotope of an "amino acid" is one in which at least one atom has been replaced with an atom having the same atomic number (number of protons) but a different mass number (the sum of the number of protons and neutrons) in an abundance ratio different from the natural abundance ratio. Examples of isotopes included in "amino acids" as used herein include hydrogen atoms, carbon atoms, nitrogen atoms, oxygen atoms, phosphorus atoms, sulfur atoms, fluorine atoms, and chlorine atoms, each of which is ２ H. ３ H. １３ C. １４ C. １５ N. １７ O. １８ O. ３２ P. ３５ S. １８ F. ３６ Cl, etc. Compounds herein containing all proportions of radioactive or non-radioactive isotopes are within the scope of the present invention.

[0067] The peptide compound according to this embodiment may contain two or more N-substituted amino acid residues. The number of N-substituted amino acid residues contained in the peptide compound according to this embodiment may be, for example, three or more, four or more, or five or more.

[0068] The peptide compound according to this embodiment may be a peptide compound having a cyclic portion (sometimes referred to herein as a "cyclic peptide compound"). As used herein, the "cyclic portion" of a peptide compound refers to a cyclic portion formed by linking two or more amino acid residues. The number of amino acid residues constituting the cyclic portion of the cyclic peptide compound may be 5 to 15, 6 to 15, 7 to 15, 8 to 15, 9 to 15, 10 to 15, 5 to 14, 6 to 14, 7 to 14, 8 to 14, 9 to 14, 10 to 14, 5 to 13, 6 to 13, 7 to 13, 8 to 13, 9 to 13, 10 to 13, 5 to 12, 6 to 12, 7 to 12, 8 to 12, 9 to 12, 10 to 12, 5 to 11, 6 to 11, 7 to 11, 8 to 11, 9 to 11, 10 to 11, or 11. The cyclic moiety is preferably formed via a covalent bond such as an amide bond, a carbon-carbon bond-forming reaction, an S-S bond, a thioether bond, or a triazole bond, and can be formed by bonding a group at the N-terminus and a group at the C-terminus of a linear peptide compound to which amino acid residues are linked. Specifically, for example, it can be formed by bonding an amino group at the N-terminus and a carboxyl group at the C-terminus of a linear peptide compound to which amino acid residues are linked. The cyclization may be performed in any manner, such as via a carbon-nitrogen bond such as an amide bond, via a carbon-oxygen bond such as an ester bond or an ether bond, via a carbon-sulfur bond such as a thioether bond, via a carbon-carbon bond, via a sulfur-sulfur bond, or via a heterocyclic ring structure. Among these, cyclization via a covalent bond such as an amide bond or a carbon-carbon bond is preferred, and cyclization via an amide bond between a carboxy group in a side chain and an amino group in the main chain is more preferred. The position of the carboxy group and amino group used for cyclization may be on the main chain or on the side chain, and is not particularly limited as long as it is in a position that allows cyclization.

[0069] The cyclic peptide compound according to this embodiment may have 15 to 46 ring atoms. As used herein, the term "number of ring atoms" refers to the number of atoms (ring atoms) in a cyclic compound, including the innermost portion of the ring. When a compound has multiple rings, the term is defined as the number of atoms in the ring with the largest number of atoms. When two rings share some atoms, the ring with the fewer shared atoms is used to calculate the number of ring atoms in each ring. To further explain the "number of ring atoms" using specific examples, for example, using this method, tetrahydrofuran (THF) has 5 ring atoms, tacrolimus (FK506) has 21 ring atoms, and Compound 1 and Compound 9 described in the Examples have 34 ring atoms.

[0070] The number of ring atoms in the cyclic peptide compound according to this embodiment may be, for example, 34 to 46, 34 to 43, 34 to 40, 34 to 37, 34 to 36, or 34. The ring atoms used to calculate the number of ring atoms may be selected from the group consisting of carbon atoms, hydrogen atoms, nitrogen atoms, oxygen atoms, sulfur atoms, phosphorus atoms, and silicon atoms, or may be selected from the group consisting of carbon atoms, hydrogen atoms, nitrogen atoms, and oxygen atoms.

[0071] A peptide compound according to one embodiment may have a ClogP / amino acid residue ratio of 1.0 or greater. Here, "number of amino acid residues" refers to the total number of amino acid residues constituting the peptide compound. For example, a cyclic peptide compound having a cyclic portion consisting of 10 amino acid residues and a linear portion consisting of 1 amino acid residue has 11 amino acid residues. The ClogP / amino acid residue ratio is a value calculated by dividing the ClogP of a peptide compound by the number of amino acid residues contained in the peptide compound. For example, if the ClogP of a peptide compound is 14.0 and the number of amino acid residues contained in the peptide compound is 7, the ClogP / amino acid residue ratio of the peptide compound is calculated to be 2.0.

[0072] The ClogP / number of amino acid residues of the peptide compound according to this embodiment may be, for example, 1.1 or more, or 1.2 or more. The upper limit of the ClogP / number of amino acid residues of the peptide compound according to this embodiment may be, for example, 1.8 or less, 1.7 or less, 1.6 or less, or 1.5 or less. Examples of ranges of the ClogP / number of amino acid residues of the peptide compound according to this embodiment include 1.0 or more and 1.8 or less, 1.0 or more and 1.7 or less, 1.1 or more and 1.6 or less, and 1.1 or more and 1.5 or less.

[0073] The compound according to this embodiment may have one of the above-described physical properties, characteristics, structure, etc., or may have any combination of two or more of them. Further specific examples of the compound according to this embodiment are described below.

[0074] The compound according to one embodiment has the following characteristics (i) and (ii): (i) a ClogP of 9 or more, and (ii) a Caco-2 Papp (cm / sec) value of 1.0E-5 or less.

[0075] The compound according to one embodiment has the following characteristics (i) and (ii): (i) ClogP is 10 or more, and (ii) Caco-2 Papp (cm / sec) is 1.0E-7 or more and 1.0E-5 or less.

[0076] The compound according to one embodiment has the following characteristics (i), (ii), and (iii): (i) a ClogP of 13 or more; (ii) a Caco-2 Papp (cm / sec) value of 1.4E-7 or more and 6.0E-6 or less; and (iii) a molecular weight of 1000 or more.

[0077] The compound according to one embodiment is a peptide compound having a cyclic portion, and has the following characteristics (i) and (ii): (i) the compound has a cyclic portion consisting of 5 to 12 amino acid residues, and the number of amino acid residues is 9 to 13. (ii) the compound has at least two N-substituted amino acid residues and at least one non-N-substituted amino acid residue.

[0078] A compound according to one embodiment is a peptide compound having a cyclic portion, and has the following characteristics (i) and (ii): (i) the compound has a cyclic portion consisting of 9 to 12 amino acid residues, and the number of amino acid residues is 9 to 13. (ii) the compound has at least three N-substituted amino acid residues and at least one non-N-substituted amino acid residue.

[0079] The compound according to one embodiment is a peptide compound having a cyclic portion, and has the following characteristics (i) and (ii): (i) the compound has a cyclic portion consisting of 11 amino acid residues, and the number of amino acid residues is 11. (ii) the compound has at least four N-substituted amino acid residues and at least one non-N-substituted amino acid residue.

[0080] A compound according to one embodiment is a peptide compound having a cyclic portion, and has the following characteristics (i), (ii), (iii), and (iv): (i) it has a cyclic portion consisting of 5 to 12 amino acid residues, and the number of amino acid residues is 9 to 13; (ii) it has at least two N-substituted amino acid residues and at least one non-N-substituted amino acid residue; (iii) it has a ClogP of 9 or more; and (iv) it has a Caco-2 Papp (cm / sec) value of 1.0E-5 or less.

[0081] A compound according to one embodiment is a peptide compound having a cyclic portion, and has the following characteristics (i), (ii), (iii), and (iv): (i) it comprises a cyclic portion consisting of 9 to 12 amino acid residues, and the number of amino acid residues is 9 to 13; (ii) it comprises at least three N-substituted amino acid residues and at least one non-N-substituted amino acid residue; (iii) it has a ClogP of 9 or more; and (iv) it has a Caco-2 Papp (cm / sec) value of 1.0E-5 or less.

[0082] The compound according to one embodiment is a peptide compound having a cyclic portion, and has the following characteristics (i), (ii), (iii), and (iv): (i) the compound has a cyclic portion consisting of 11 amino acid residues, and the number of amino acid residues is 11; (ii) the compound has at least four N-substituted amino acid residues and at least one non-N-substituted amino acid residue; (iii) the ClogP is 11 or more; and (iv) the Caco-2 Papp (cm / sec) value is 6.0E-6 or less.

[0083] A compound according to one embodiment is a peptide compound having a cyclic portion, and has the following characteristics (i) and (ii): (i) the number of amino acid residues is 9 to 13, and the number of ring atoms in the cyclic portion is 34. (ii) the compound contains at least two N-substituted amino acid residues and at least one non-N-substituted amino acid residue.

[0084] A compound according to one embodiment is a peptide compound having a cyclic portion, and has the following characteristics (i), (ii), (iii), (iv), and (v): (i) the number of amino acid residues is 9 to 13, and the number of ring atoms in the cyclic portion is 34; (ii) the compound contains at least two N-substituted amino acid residues and at least one non-N-substituted amino acid residue; (iii) the ClogP is 9 or greater; (iv) the Caco-2 Papp (cm / sec) value is 1.0E-5 or less; and (v) the molecular weight is 1,000 or greater.

[0085] A compound according to one embodiment is a peptide compound having a cyclic portion, and has the following characteristics (i), (ii), (iii), (iv), and (v): (i) the number of amino acid residues is 11, and the number of ring atoms in the cyclic portion is 34. (ii) the compound contains at least four N-substituted amino acid residues and at least one non-N-substituted amino acid residue. (iii) the ClogP is 13 or greater. (iv) the Caco-2 Papp (cm / sec) value is 1.4E-7 or greater and 6.0E-6 or less. (v) the molecular weight is 1,000 or greater.

[0086] The compound according to this embodiment may not include, for example, cyclosporine and its analogs. A cyclosporine analog refers to a chemically modified cyclosporine that maintains the biological activity of cyclosporine. An example of a cyclosporine analog is dihydrocyclosporine D.

[0087] Compounds according to this embodiment can also be used, for example, (5S,8S,11S,15S,18S,23aS,29S,35S,37aS)-8-((S)-sec-butyl)-18-cyclopentyl-29-(3,5-difluoro-4-(trifluoromethyl)phenethyl)-36-ethyl-11-isobutyl-N,N,5,6,12,16,19,33-octamethyl-35-(4-methylbenzyl)-4,7,10,13,17,20,23,28,31,34,37-undecaoxotetratriacontahydro-2H,4H-spiro[azeto[ 2,1-u]pyrrolo[2,1-i][1,4,7,10,13,16,19,22,25,28,31]undecaazacyclotetratriacontin-21,1'-cyclopentane]-15-carboxamide, (5S,8S,11S,15S,18S,23aS,25R,29S,35S,37aS)-8-((S)-sec-butyl)-35-(cyclohexylmethyl)-18-cyclopentyl-29-(3,5-difluoro-4-(trifluoromethyl)phenethyl)-25-ethoxy-11-isobutyl-N,N,5,6,12,16 , 19,33,36-nonamethyl-4,7,10,13,17,20,23,28,31,34,37-undecaoxotetratriacontahydro-2H,4H-spiro[azeto[2,1-u]pyrrolo[2,1-i][1,4,7,10,13,16,19,22,25,28,31]undecaazacyclotetratriacontin-21,1'-cyclopentane]-15-carboxamide, (3S,9S,18S,21S,25S,28S,34S)-3-[2-[3-chloro-4-(trifluoromethyl)phenyl]ethyl]-28-cyclopentane Cyclohexyl-9-(cyclohexylmethyl)-21-isobutyl-7,10,13,16,22,26,29-heptamethyl-18-[(1S)-1-methylpropyl]-25-(piperidine-1-carbonyl)spiro[1,4,7,10,13,16,19,22,26,29,32-undecazabicyclo[32.3.0]heptatriacontane-31,1'-cyclopentane]-2,5,8,11,14,17,20,23,27,30,33-undecaone, (5S,8S,11S,15S,18S,23aS,29S,35S,37aS)-8-((S)-sec-butyl)-29-(3-chloro-4-(trifluoromethyl)phenethyl)-35-(cyclohexylmethyl)-18-cyclopentyl-11-isobutyl-5,6,12,16,19,33,36-heptamethyl-15-(morpholine-4-carbonyl)docosahydro-2H,4H-spiro[azeto[ 2,1-u]pyrrolo[2,1-i][1,4,7,10,13,16,19,22,25,28,31]undecaazacyclotetratriacontin-21,1′-cyclopentane]-4,7,10,13,17,20,23,28,31,34,37(14H,22H)-undecaone, and (5S,8S,11S,15S,18S,23aS , 29S, 35S, 37aS)-8-((S)-sec-butyl)-29-(3-chloro-4-(trifluoromethyl)phenethyl)-18-cyclopentyl-36-ethyl-11-isobutyl-5,6,12,16,19,33-hexamethyl-35-(4-methylbenzyl)-15-(morpholine-4-carbonyl)docosahydro-2H , 4H-spiro[azeto[2,1-u]pyrrolo[2,1-i][1,4,7,10,13,16,19,22,25,28,31]undecaazacyclotetratriacontin-21,1'-cyclopentane]-4,7,10,13,17,20,23,28,31,34,37(14H,22H)-undecaone.

[0088] Generally, only one compound according to this embodiment is contained in one formulation.

[0089] (Oil component containing polyoxyethylene structure) The preparation according to this embodiment contains an oil component containing a polyoxyethylene structure (sometimes simply referred to as "oil component"). The polyoxyethylene structure is a -(CH ２ CH ２ O) ｎ - (n is a natural number).

[0090] The average number of moles of ethylene oxide added in the oil component according to this embodiment may be, for example, 2 or more and 150 or less. The average number of moles of ethylene oxide added in the oil component according to this embodiment may be, for example, 5 or more, 10 or more, 15 or more, 20 or more, 25 or more, or 30 or more, and may be 140 or less, 130 or less, 120 or less, 110 or less, 100 or less, 90 or less, 80 or less, 70 or less, 60 or less, 50 or less, 40 or less, 39 or less, 38 or less, 37 or less, or 36 or less. Examples of ranges for the average number of moles of ethylene oxide added in the oil component according to this embodiment include 10 or more and 90 or less, 15 or more and 60 or less, 20 or more and 50 or less, 25 or more and 40 or less, 30 or more and 39 or less, or 35.

[0091] Examples of the oil component according to this embodiment include polyoxyethylene castor oil, polyoxyethylene hydrogenated castor oil, and polyoxyethylene sorbitan fatty acid ester. More specific examples of the oil component according to this embodiment include polyoxyethylene castor oil having an average number of moles of ethylene oxide added of 30 to 39, and polyoxyethylene castor oil having an average number of moles of ethylene oxide added of 35.

[0092] The oily component according to the present embodiment may be used alone or in combination of two or more.

[0093] (Preparation) The preparation according to this embodiment contains at least the compound described above and an oily component containing a polyoxyethylene structure. The content of the oily component containing a polyoxyethylene structure in 100% by volume of the preparation according to this embodiment may be 2.0% by volume or more and 7.5% by volume or less.

[0094] The content of the oily component containing a polyoxyethylene structure in 100% by volume of the preparation according to this embodiment is, for example, 2.1% by volume or more, 2.2% by volume or more, 2.3% by volume or more, 2.4% by volume or more, 2.5% by volume or more, 2.6% by volume or more, 2.7% by volume or more, 2.8% by volume or more, 2.9% by volume or more, 3.0% by volume or more, 3.1% by volume or more, 3.2% by volume or more, 3.3% by volume or more, 3.4% by volume or more, 3.5% by volume or more, 3.6% by volume or more, 3.7% by volume or more, 3.8% by volume or more, 3.9% by volume or more, 4.0% by volume or more, 4.1% by volume or more, 4.2% by volume or more, 4.3% by volume or more, 4.4% by volume or more, 4.5% by volume or more. or more, 4.6 vol% or more, 4.7 vol% or more, 4.8 vol% or more, 4.9 vol% or more, 5.0 vol% or more, 5.1 vol% or more, 5.2 vol% or more, 5.3 vol% or more, 5.4 vol% or more, 5.5 vol% or more, 5.6 vol% or more, 5.7 vol% or more, 5.8 vol% or more, 5.9 vol% or more, 6.0 vol% or more, 6.1 vol% or more, 6.2 vol% or more, 6.3 vol% or more, 6.4 vol% or more, 6.5 vol% or more, 6.6 vol% or more, 6.7 vol% or more, 6.8 vol% or more, 6.9 vol% or more, 7.0 vol% or more, 7.1 vol% or more, 7.2 vol% or more, 7.3 vol% or more, or 7.4 vol% or more.

[0095] The content of the oily component containing a polyoxyethylene structure in 100% by volume of the preparation according to this embodiment is, for example, 7.4% by volume or less, 7.3% by volume or less, 7.2% by volume or less, 7.1% by volume or less, 7.0% by volume or less, 6.9% by volume or less, 6.8% by volume or less, 6.7% by volume or less, 6.6% by volume or less, 6.5% by volume or less, 6.4% by volume or less, 6.3% by volume or less, 6.2% by volume or less, 6.1% by volume or less, 6.0% by volume or less, 5.9% by volume or less, 5.8% by volume or less, 5.7% by volume or less, 5.6% by volume or less, 5.5% by volume or less, 5.4% by volume or less, 5.3% by volume or less, 5.2% by volume or less, 5.1% by volume or less, 5.0% by volume vol.%, 4.9 vol.% or less, 4.8 vol.% or less, 4.7 vol.% or less, 4.6 vol.% or less, 4.5 vol.% or less, 4.4 vol.% or less, 4.3 vol.% or less, 4.2 vol.% or less, 4.1 vol.% or less, 4.0 vol.% or less, 3.9 vol.% or less, 3.8 vol.% or less, 3.7 vol.% or less, 3.6 vol.% or less, 3.5 vol.% or less, 3.4 vol.% or less, 3.3 vol.% or less, 3.2 vol.% or less, 3.1 vol.% or less, 3.0 vol.% or less, 2.9 vol.% or less, 2.8 vol.% or less, 2.7 vol.% or less, 2.6 vol.% or less, 2.5 vol.% or less, 2.4 vol.% or less, 2.3 vol.% or less, 2.2 vol.% or less, or 2.1 vol.% or less.

[0096] The range of the content of the oily component containing a polyoxyethylene structure in 100% by volume of the preparation according to this embodiment is, for example, 2.1% by volume or more and 7.4% by volume or less, 2.2% by volume or more and 7.3% by volume or less, 2.3% by volume or more and 7.2% by volume or less, 2.4% by volume or more and 7.1% by volume or less, 2.5% by volume or more and 7.0% by volume or less, 2.6% by volume or more and 6.9% by volume or less, 2.7% by volume or more and 6.8% by volume or less, 2.8% by volume or more and 6.7% by volume or less, 2.9% by volume or more and 6.6% by volume or less, 3.0% by volume or more and 6.5% by volume or less, 3.1% by volume or more and 6.4% by volume or less, 3.2% by volume or more and 6.3% by volume or less, 3.3% by volume or more and 6.2% by volume or less, 3.4% by volume or more and 6.1% by volume or less, 3.5% by volume or more and 6.6 ... Examples of the concentration include 2.0 vol% or more and 3.9 vol% or less, 2.0 vol% or more and 3.9 vol% or less, 2.0 vol% or more and 3.5 vol% or less, 4.1 vol% or more and 4.9 vol% or less, 5.1 vol% or more and 7.5 vol% or less, 5.5 vol% or more and 7.5 vol% or less, and 6.0 vol% or more and 7.5 vol% or less. Further examples of the range of the content of the oily component containing a polyoxyethylene structure in 100% by volume of the preparation according to this embodiment include 2.0% by volume or more and less than 7.5% by volume, 2.0% by volume or more and 7.0% by volume or less, 2.0% by volume or more and 5.0% by volume or less, 2.5% by volume or more and 7.5% by volume or less, and 2.5% by volume or more and 5.0% by volume or less.

[0097] The content of the compound in the formulation according to this embodiment is not particularly limited, and may be, for example, 0.1 mg / mL or more and 100 mg / mL or less. The content of the compound in the formulation according to this embodiment may be, for example, 0.2 mg / mL or more, 0.3 mg / mL or more, 0.4 mg / mL or more, 0.5 mg / mL or more, 0.6 mg / mL or more, 0.7 mg / mL or more, 0.8 mg / mL or more, 0.9 mg / mL or more, or 1 mg / mL or more, or 90 mg / mL or less, 80 mg / mL or less, 70 mg / mL or less, 60 mg / mL or less, 50 mg / mL or less, 40 mg / mL or less, or 30 mg / mL or less. Examples of ranges of the content of the compound in the formulation according to this embodiment include 0.2 mg / mL to 90 mg / mL, 0.3 mg / mL to 80 mg / mL, 0.4 mg / mL to 70 mg / mL, 0.5 mg / mL to 60 mg / mL, 0.6 mg / mL to 50 mg / mL, 0.7 mg / mL to 40 mg / mL, and 0.8 mg / mL to 30 mg / mL.

[0098] The formulation of this embodiment may include a carrier, such as an aqueous medium such as saline, buffered saline, water, and isotonic aqueous buffer, dimethyl sulfoxide (DMSO), and combinations thereof.

[0099] When the formulation according to this embodiment contains DMSO, the content of DMSO in 100% by volume of the formulation may be, for example, 1% by volume or more and 30% by volume or less, 3% by volume or more and 25% by volume or less, 5% by volume or more and 20% by volume or less, 7% by volume or more and 15% by volume or less, or 10% by volume.

[0100] The formulation according to this embodiment may contain other components (other components) in addition to the above-described compounds and the oily component containing a polyoxyethylene structure, as long as the effects of the present invention are not impaired. Examples of other components include surfactants other than the oily component containing a polyoxyethylene structure (e.g., lauroyl-L-carnitine), stabilizers, preservatives, antioxidants, disintegrants, excipients, binders, glidants, lubricants, etc.

[0101] The formulation according to this embodiment contains a compound and an oily component containing a polyoxyethylene structure at a specific concentration, thereby generally enhancing the oral absorbability of the compound. Therefore, the formulation according to this embodiment may not contain any surfactant other than the oily component containing a polyoxyethylene structure. More specifically, the formulation according to this embodiment may not contain lauroyl-L-carnitine.

[0102] The components contained in the formulation of this embodiment may consist essentially of water, a compound, an oily component containing a polyoxyethylene structure, and DMSO, or may consist solely of water, a compound, an oily component containing a polyoxyethylene structure, and DMSO. In this specification, "substantially" means that the listed components (such as, but not limited to, water, a compound, an oily component containing a polyoxyethylene structure, and DMSO) are the main components, and other components may be included as long as they do not negatively affect the effects of one aspect of the present invention, or in amounts or forms that do not negatively affect the effects of one aspect of the present invention. For example, components not listed therein (e.g., ethanol, etc.) may be included as long as they do not negatively affect the effects of one aspect of the present invention, or in amounts or forms that do not negatively affect the effects of one aspect of the present invention.

[0103] The dosage form of the preparation according to this embodiment is not particularly limited, but is typically a liquid preparation.

[0104] The formulation according to this embodiment is preferably an oral formulation since the oral absorbability of the compound is enhanced.

[0105] The subject to which the formulation of this embodiment is administered is not particularly limited and may be a human or a non-human animal. Examples of non-human animals include dogs, monkeys, minipigs, rabbits, rats, and mice. The subject is preferably a non-human animal, more preferably a dog, monkey, minipigs, rabbits, rats, and mice, even more preferably a rat and mouse, and even more preferably a mouse.

[0106] The method of administering the formulation according to this embodiment is not particularly limited, and may be oral or parenteral administration, but oral administration is preferred because the formulation according to this embodiment has enhanced oral absorbability of the compound.

[0107] The dosage of the formulation according to this embodiment is not particularly limited, and may be administered so that the dosage of the compound per kg of subject body weight is 0.1 mg / kg to 1000 mg / kg. The dosage of the compound may be, for example, 1 mg / kg to 500 mg / kg, 1 mg / kg to 100 mg / kg, 1 mg / kg to 50 mg / kg, 3 mg / kg to 30 mg / kg, 10 mg / kg to 30 mg / kg, 0.1 mg / kg to 10 mg / kg, 1 mg / kg to 5 mg / kg, 10 mg / kg to 100 mg / kg, 15 mg / kg to 50 mg / kg, 20 mg / kg to 40 mg / kg, 25 mg / kg to 35 mg / kg, 3 mg / kg, or 30 mg / kg.

[0108] The formulation according to this embodiment comprises blending a compound and an oily component containing a polyoxyethylene structure, and the oily component containing a polyoxyethylene structure is blended so that its content in 100% by volume of the formulation is 2.0% by volume or more and 7.5% by volume or less.

[0109] More specifically, the production method may include mixing an aqueous medium such as water and / or a carrier such as DMSO with the compound and an oily component containing a polyoxyethylene structure by stirring, etc. Furthermore, if necessary, the above-mentioned other components may be further added.

[0110] In the formulation according to this embodiment, the oily component containing a polyoxyethylene structure can be considered to have the effect of promoting the absorption of the compound, and can be considered an active ingredient capable of promoting the absorption of the compound. Similarly, the oily component containing a polyoxyethylene structure can be considered to have the effect of improving the solubility of the compound, and can be considered an active ingredient capable of improving the solubility of the compound. Therefore, as one aspect of the present invention, there is provided an absorption enhancer for a compound, which comprises an oily component containing a polyoxyethylene structure as an active ingredient, and the content of the oily component in a 100% by volume formulation containing the compound is 2.0% by volume or more and 7.5% by volume or less. Similarly, as another aspect of the present invention, there is provided a solubility enhancer for a compound, which comprises an oily component containing a polyoxyethylene structure as an active ingredient, and the content of the oily component in a 100% by volume formulation containing the compound is 2.0% by volume or more and 7.5% by volume or less.

[0111] From the above viewpoint, the preparation of this embodiment is preferably that the absorption of compound is promoted compared with the preparation that does not contain the oily component that contains polyoxyethylene structure.In addition, the preparation of this embodiment is preferably that the solubility of compound is improved compared with the preparation that does not contain the oily component that contains polyoxyethylene structure.Here, the "preparation that does not contain the oily component that contains polyoxyethylene structure" to be compared refers to the preparation that has a composition in which the oily component that contains polyoxyethylene structure is replaced with a carrier (for example, water) in the preparation of this embodiment.

[0112] The formulation according to this embodiment has a general purpose, enhanced oral absorption of compounds, and can therefore be suitably used for screening candidate compounds. Candidate compounds include, in particular, pharmaceutical candidate compounds (clinical candidate compounds). A method for screening candidate compounds using the formulation according to this embodiment will now be described.

[0113] [Candidate Compound Screening Method] The candidate compound screening method according to this embodiment (hereinafter also simply referred to as the "screening method") comprises administering the above-described formulation according to the present invention to a subject. In the following description, the compound contained in the formulation may be particularly referred to as the "test compound."

[0114] The screening method according to this embodiment may include selecting a candidate compound using at least one index selected from the group consisting of the following (1), (2), and (3): (1) efficacy, (2) toxicity, and (3) pharmacokinetic properties.

[0115] In the screening method according to this embodiment, the efficacy and / or toxicity of a test compound administered to a subject can be measured, and test compounds with recognized or high efficacy can be selected, or test compounds with low or no toxicity can be selected. The efficacy and / or toxicity of a test compound can be evaluated, for example, by comparing it with a preset threshold value. Alternatively, similar measurements can be performed using a compound with efficacy as a control, and test compounds with higher efficacy or lower toxicity than the control can be selected. Examples of efficacy include, but are not limited to, cell proliferation inhibition, cytotoxicity, and tumor growth inhibition. Examples of toxicity include, but are not limited to, hepatotoxicity, nephrotoxicity, and cardiotoxicity.

[0116] Furthermore, in the screening method according to this embodiment, the pharmacokinetic properties of a test compound can also be evaluated by measuring the blood concentration of the test compound in a subject administered with the test compound. Here, "pharmacokinetic properties" refers to properties such as the effective blood concentration, blood half-life, and / or elimination rate of the test compound in the subject's body. The pharmacokinetic properties of a test compound may be evaluated, for example, by comparison with a preset threshold value. For example, a substance with a higher effective blood concentration, a longer blood half-life, or a slower elimination rate is generally considered to have better pharmacokinetic properties, but this is not a limitation. Furthermore, the method for measuring the blood concentration of a test compound is not particularly limited.

[0117] The screening method according to this embodiment may include (3) selecting a candidate compound using pharmacokinetic properties as an index.

[0118] The screening method according to this embodiment may select a candidate compound from a plurality of test compounds. In this case, each of the plurality of test compounds is administered to a subject as a separate formulation. The number of test compounds is not particularly limited, and may be, for example, 2 or more, 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, or 8 or more. Note that since screening involves selecting compounds that meet certain conditions, the screening method according to this embodiment can also be performed on a single test compound.

[0119] In the screening method according to this embodiment, the subject to which the formulation according to the present invention is administered is not particularly limited, and may be a non-human animal. Examples of the subject include dogs, monkeys, minipigs, rabbits, rats, and mice. Rats and mice are preferred, and mice are more preferred.

[0120] In the screening method according to this embodiment, the method of administering the formulation according to the present invention is not particularly limited, and may be orally or parenterally. However, the formulation according to the present invention is preferably administered orally because the oral absorbability of the compound is enhanced.

[0121] In the screening method according to this embodiment, the dosage of the formulation according to the present invention is not particularly limited, and may be administered so that the dose of the test compound per kg of subject body weight (kg) is 0.1 mg / kg to 1000 mg / kg. The dose of the test compound may be, for example, 1 mg / kg to 500 mg / kg, 1 mg / kg to 100 mg / kg, 1 mg / kg to 50 mg / kg, 3 mg / kg to 30 mg / kg, 10 mg / kg to 30 mg / kg, 0.1 mg / kg to 10 mg / kg, 1 mg / kg to 5 mg / kg, 10 mg / kg to 100 mg / kg, 15 mg / kg to 50 mg / kg, 20 mg / kg to 40 mg / kg, 25 mg / kg to 35 mg / kg, 3 mg / kg, or 30 mg / kg.

[0122] [Method for comparing test compounds] The method according to this embodiment is a method for comparing a plurality of test compounds, and includes administering the above-described formulation according to the present invention to a subject, and comparing the plurality of test compounds using at least one index selected from the group consisting of the following (1), (2), and (3): (1) efficacy, (2) toxicity, and (3) pharmacokinetic properties.

[0123] In the comparison method according to this embodiment, the efficacy and / or toxicity of test compounds administered to a subject are measured and compared between test compounds. This allows, for example, the selection of test compounds with relatively high efficacy or relatively low toxicity. Alternatively, similar measurements can be performed using a control compound with efficacy as the test compound to select test compounds with higher efficacy or lower toxicity compared to the control. Examples of efficacy include, but are not limited to, cell proliferation inhibitory activity, cytotoxic activity, and tumor growth inhibitory activity. Examples of toxicity include, but are not limited to, hepatotoxicity, nephrotoxicity, and cardiotoxicity.

[0124] Furthermore, in the comparison method according to this embodiment, the pharmacokinetic properties of a test compound can be evaluated by measuring the blood concentration of the test compound in a subject administered the test compound. Here, "pharmacokinetic properties" refers to the concentration of the test compound in the subject's body, preferably characteristics such as the effective blood concentration, blood half-life, and / or elimination rate of the test compound in the subject's body. By comparing the pharmacokinetic properties of test compounds, test compounds with relatively superior pharmacokinetic properties can be selected. Alternatively, similar measurements can be performed using a control compound with pharmacological activity as the test compound to select test compounds with superior pharmacokinetic properties compared to the control. For example, generally, the higher the effective blood concentration, the longer the blood half-life, or the slower the elimination rate, the better the pharmacokinetic properties of the substance, but this is not a limitation. Furthermore, the method for measuring the blood concentration of the test compound is not particularly limited.

[0125] The comparison method according to this embodiment may include (3) comparing test compounds using pharmacokinetic properties as an index. The number of test compounds is not particularly limited, and may be, for example, two or more, or three or more.

[0126] In the comparative method according to this embodiment, the subject to which the formulation according to the present invention is administered is not particularly limited, and may be a non-human animal. Examples of the subject include dogs, monkeys, minipigs, rabbits, rats, and mice. Rats and mice are preferred, and mice are more preferred.

[0127] In the comparative method according to this embodiment, the method of administering the formulation according to the present invention is not particularly limited, and may be orally or parenterally. On the other hand, the formulation according to the present invention is preferably administered orally because the oral absorption of the compound is enhanced.

[0128] In the comparative method according to this embodiment, the dosage of the formulation according to the present invention is not particularly limited, and may be administered so that the dose of the test compound per kg of subject body weight (kg) is 0.1 mg / kg to 1000 mg / kg. The dose of the test compound may be, for example, 1 mg / kg to 500 mg / kg, 1 mg / kg to 100 mg / kg, 1 mg / kg to 50 mg / kg, 3 mg / kg to 30 mg / kg, 10 mg / kg to 30 mg / kg, 0.1 mg / kg to 10 mg / kg, 1 mg / kg to 5 mg / kg, 10 mg / kg to 100 mg / kg, 15 mg / kg to 50 mg / kg, 20 mg / kg to 40 mg / kg, 25 mg / kg to 35 mg / kg, 3 mg / kg, or 30 mg / kg.

[0129] [Method for enhancing systemic absorption of a test compound] The method according to this embodiment is a method for enhancing systemic absorption of a test compound in a subject, and includes administering the above-described formulation according to the present invention to the subject. The systemic absorption may be gastrointestinal absorption. The formulation according to the present invention has excellent oral absorption of the compound, and therefore can be suitably used in a method for enhancing systemic absorption of a test compound in a subject.

[0130] The present invention will be described in more detail below with reference to examples, but the present invention is not limited to the following examples. All starting materials and reagents were obtained from commercial suppliers or synthesized using known methods.

[0131] The following abbreviations are used in the examples: BF ３ OEt ２: Boron trifluoride diethyl ether complex COMU: (1-cyano-2-ethoxy-2-oxoethylideneaminooxy)dimethylaminomorpholinocarbenium hexafluorophosphate DBU: 1,8-diazabicyclo[5.4.0]-7-undecene DCM: dichloromethane DCE: 1,2-dichloroethane DIAD: diisopropyl azodicarboxylate DIC: N,N'-diisopropylcarbodiimide DIPEA: N,N-diisopropylethylamine DMA: dimethylacetamide DMAP: N,N-dimethyl-4-aminopyridine DMF: N,N-dimethylformamide DMSO: dimethyl sulfoxide dtbbpy: 4,4'-di-tert-butyl-2,2'-bipyridyl EDTA: ethylenediaminetetraacetic acid EtOAc: ethyl acetate FA: formic acid Fmoc: 9-fluorenylmethyloxycarbonyl group Fmoc-OSu: N-(9-fluorenylmethyloxycarbonyloxy)succinimide HATU: O-(7-aza-1H-benzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate HOBt: 1-hydroxybenzotriazole HBTU: O-(1H-benzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate HMDS: 1,1,1,3,3,3-hexamethyldisilazane H-SAR-OTBU HCl: sarcosine tert-butyl ester hydrochloride IPAC: isopropyl acetate MeOH: methyl alcohol Ns: 2-nitrobenzenesulfonyl group Oxyma: ethyl cyano(hydroxyimino)acetate pip: piperidine TBME: t-butyl methyl ether TES: triethylsilane TfOH: trifluoromethanesulfonic acid TMSOTf: trimethylsilyl trifluoromethanesulfonate THF: tetrahydrofuran TsOH: p-toluenesulfonic acid THP: tetrahydropyranyl group Trt: triphenylmethyl group WSC.HCl, WSCI: 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride

[0132] Example 1 Synthesis of Cyclic Peptide Compounds Cyclic peptide compounds 1 to 10 (also simply referred to as compounds 1 to 10) shown in Table 2 were prepared by peptide elongation using the Fmoc peptide synthesis method described in WO 2013 / 100132 or WO 2018 / 225864, following the basic route below. Specifically, the process involves five steps: 1) a peptide elongation reaction using the Fmoc method from the N-terminus of an amino acid, with the carboxylic acid of the Asp side chain supported on a 2-chlorotrityl resin; 2) a process of cleaving the peptide from the 2-chlorotrityl resin; 3) an amide cyclization reaction involving condensation of the carboxylic acid of the Asp side chain, which is released from the 2-chlorotrityl resin during the cleavage process, with the amino group at the N-terminus of the peptide chain; 4) deprotection of the protecting groups of the side chain functional groups contained in the peptide chain; and 5) purification of the compound by preparative HPLC. Additionally, all starting materials and reagents were obtained from commercial suppliers or synthesized using known methods.

[0133] Synthesis of (S)-3-((((9H-fluoren-9-yl)methyloxy)carbonyl)amino)-4-oxo-4-(piperidin-1-yl)butanoic acid-2-chlorotriethyl resin (Fmoc-Asp(O-Trt(2-Cl)-Resin)-pip) It was synthesized from (S)-3-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-oxo-4-(piperidin-1-yl)butanoic acid by the method described in WO 2013 / 100132.

[0134] Synthesis of Compound 5 (S)-3-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-oxo-4-(piperidin-1-yl)butanoate-2-chlorotrityl resin (Fmoc-Asp(O-Trt(2-Cl)-Resin)-pip) (0.269 mmol / g, 100 mg, 0.0269 mmol) was used to prepare Fmoc-MeAla-OH, Fmoc-MePhe-OH, Fmoc-Ser(tBu)-OH, Fmoc-MeAbu-OH, Fmoc-MeGly-OH, Fmoc-Th The above-mentioned peptide elongation reaction using r(THP)-OH, Fmoc-MeLeu-OH, Fmoc-Ala(3-Bzt)-OH, and Fmoc-D-Leu-OH, followed by cleavage of the elongated peptide from the resin, cyclization of the cleaved peptide (using o-(7-aza-1H-benzotriazol-1-yl)-N,N,N,N-tetramethyluronium hexafluorophosphate (HATU) as a cyclization reagent), and purification of the cyclic peptide compound were carried out to obtain the target compound 5 (29.72 mg).

[0135] Synthesis of Compound 9

[0136] Fmoc-Asp(OAl)-OH (Compound 11a, (2S)-2-(9H-fluoren-9-ylmethoxycarbonylamino)-4-oxo-4-prop-2-enoxybutanoic acid, CAS number 146982-24-3) (200 g, 506 mmol), p-toluenesulfonic acid monohydrate (5.7 g, 0.05 equivalents), and paraformaldehyde (45.6 g, 3 equivalents) were mixed in toluene (2000 mL) and stirred at 110°C for 16 hours. The solvent was evaporated from the reaction solution under reduced pressure, and the residue was dissolved in ethyl acetate and washed twice with an aqueous sodium bicarbonate solution. The organic layer was dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure. The residue was purified by silica gel column chromatography (ethyl acetate / petroleum ether, 0 / 100 to 30 / 70) to obtain compound 11b (9H-fluoren-9-ylmethyl (4S)-5-oxo-4-(2-oxo-2-prop-2-enoxyethyl)-1,3-oxazolidine-3-carboxylate) (175 g, 85%). This was combined with another batch synthesized in the same manner and used in the next reaction. LCMS (ESI) m / z = 408 (M+H) + Retention time: 1.407 min (Analysis condition: SMDmethod_20).

[0137] A mixed solution of compound 11b (100 g, 245 mmol), zinc bromide (ZnBr2) (110 g, 496 mmol), and TES (56 g, 481.6 mmol) in DCM (1 L) was stirred under a nitrogen atmosphere at room temperature for 48 hours. Four batches of the same scale reaction mixture were combined, and the solvent was evaporated under reduced pressure. The residue was dissolved in TBME and extracted ten times with 0.5 M phosphate buffer (pH = approximately 7.5). The aqueous layers were combined, adjusted to pH 2 with 5 N hydrochloric acid, and extracted twice with isopropyl acetate (IPAC). The organic layers were combined, dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure. To remove IPAC, TBME was added to the resulting residue and the solvent was distilled off under reduced pressure six times to give compound 11c ((2S)-2-[9H-fluoren-9-ylmethoxycarbonyl(methyl)amino]-4-oxo-4-prop-2-enoxybutanoic acid) (270 g, 54%). LCMS (ESI) m / z = 410 (M + H) + retention time: 1.956 minutes (analysis condition SMDmethod_05).

[0138] WSCI·HCl (0.506 g, 2.64 mmol) was dissolved in DMF (4.4 mL), and HOBt (0.356 g, 2.64 mmol) and compound 11c ((2S)-2-[9H-fluoren-9-ylmethoxycarbonyl(methyl)amino]-4-oxo-4-prop-2-enoxybutanoic acid, Fmoc-MeAsp(OAl)-OH) (0.9 g, 2.2 mmol) were added, followed by stirring at 0° C. for 10 minutes. Morpholine (0.23 mL, 2.64 mmol) was added dropwise to the reaction solution, followed by stirring at 0° C. for 1 hour. Ethyl acetate (9 mL) was added to the reaction mixture, which was washed with 0.5 N hydrochloric acid, water, saturated aqueous sodium bicarbonate / water (1 / 1), and saturated saline / water (1 / 1). After drying over anhydrous sodium sulfate, the solvent was evaporated under reduced pressure to give Compound 11d as a crude product (522 mg, 50%). LCMS (ESI) m / z = 479 (M + H) + retention time: 0.87 minutes (analysis condition SQDFA05).

[0139] Under a nitrogen atmosphere, DBU (3.15 mL, 20.90 mmol) was added dropwise to a solution of compound 11d (10 g, 20.90 mmol) in anhydrous DMF (50 mL) at room temperature and stirred for 10 minutes. Pyridine hydrochloride (2.66 g, 22.99 mmol) was added to the reaction mixture at room temperature and stirred for 10 minutes. A separately prepared active ester solution (described below) was added to the reaction mixture at room temperature, followed by the dropwise addition of DIPEA (4.01 mL, 22.99 mmol). The resulting reaction mixture was stirred at room temperature for 4 hours and 15 minutes, after which ethyl acetate (100 mL), hexane (20 mL), and 1N hydrochloric acid (100 mL) were added. The resulting mixture was extracted with ethyl acetate-hexane (5:1) (total organic phase: approximately 300 mL). The organic phase was washed with 1N hydrochloric acid (100 mL), water (100 mL), aqueous sodium bicarbonate solution (100 mL x 2), and brine (100 mL). After drying over sodium sulfate, the solvent was evaporated under reduced pressure. The resulting crude product was purified by normal phase silica gel chromatography (hexane-ethyl acetate). This was dissolved in TBME (100 mL) and washed with aqueous potassium carbonate solution (1 w / v%, 50 mL). The organic phase was washed with saturated brine / water (1 / 1, 50 mL), dried over sodium sulfate, and the solvent was evaporated under reduced pressure to obtain Compound 11e (9.98 g, 81%).

[0140] An activated ester solution was prepared as follows. Under a nitrogen atmosphere, anhydrous DMF (55 mL) was added to a mixture of (αS)-α-[[(9H-fluoren-9-ylmethoxy)carbonyl]methylamino]cyclopentaneacetic acid (7.53 g, 19.85 mmol), WSCI·HCl (5.21 g, 27.2 mmol), and Oxyma (3.56 g, 25.08 mmol) at room temperature, and the resulting reaction mixture was stirred for 40 minutes. This reaction mixture was used as an activated ester solution. LCMS (ESI) m / z = 618 (M+H)+ Retention time: 0.96 minutes (Analysis condition SQDFA05).

[0141] Under a nitrogen atmosphere, anhydrous DCM (15 mL) was added to a mixture of compound 11e (9.90 g, 16.03 mmol) and tetrakis(triphenylphosphine)palladium(0) (0.185 g, 0.16 mmol), and the mixture was stirred at room temperature. Phenylsilane (1.38 mL, 11.22 mmol) was added dropwise to the resulting solution. After stirring the resulting reaction mixture for 30 minutes, TBME (100 mL) was added, followed by the slow dropwise addition of aqueous sodium bicarbonate (a two-fold dilution of a saturated aqueous solution) (100 mL) to quench the reaction. The resulting mixture was extracted twice with water (total aqueous phase volume approximately 150 mL), and DCM (100 mL) and phosphoric acid (5.6 mL) were added to the aqueous phase. The resulting mixture was extracted twice with DCM (total organic phase volume: approximately 200 mL), and the organic phase was washed with brine (80 mL × 2) and dried over sodium sulfate. The solvent was then evaporated under reduced pressure to give Compound 11f (9.57 g, quant.), which was used in the next reaction. LCMS (ESI) m / z = 578 (M + H) + Retention time: 0.79 min (Analysis condition SQDFA05).

[0142] 2-Chlorotrityl chloride resin (1.36 mmol / g, 20.5 g, 15.07 mmol) and DCM (140 mL) were placed in a filter-equipped reaction vessel (400 mL) and allowed to stand at room temperature for 1 hour. After the DCM was removed from the reaction vessel using nitrogen pressure, a DCM (140 mL) solution containing compound 11f (9.50 g, 16.45 mmol), methanol (5.32 mL, 132 mmol), and DIPEA (13.8 mL, 79 mmol) was added to the reaction vessel, and the reaction vessel was shaken at 25°C and 60 rpm for 60 minutes. After the reaction solution was removed from the reaction vessel using nitrogen pressure, a DCM (140 mL) solution of methanol (20 mL, 493 mmol) and DIPEA (13.8 mL, 79 mmol) was added to the reaction vessel, and the reaction vessel was shaken at 25°C and 60 rpm for 60 minutes. The reaction mixture was removed under nitrogen pressure, and then DCM (140 mL) was added and mixed, followed by removal under nitrogen pressure. This washing procedure of the resin with DCM was repeated five times, and the resulting resin was dried under reduced pressure for one day to obtain Compound 11 (26.89 g).

[0143] The amount of amino acid supported on the resin was calculated as follows, with reference to the method described in the literature (Letters in Peptide Science, 2002, 9, 203). The obtained compound 11 (10 mg) was placed in a reaction vessel, DMF (2 mL) was added, and the mixture was left to stand at room temperature for 1 hour. Then, DBU (40 μL) was added to the reaction vessel and the mixture was shaken at 25°C for 30 minutes. Then, DMF (8 mL) was added to the reaction mixture, and 1 mL of the resulting solution was diluted with DMF (11.5 mL). The absorbance (294 nm) of the resulting diluted solution was measured (measured using a Shimadzu UV-1600PC (cell length 1.0 cm)), and the amount of compound 11 supported was calculated to be 0.415 mmol / g.

[0144]

[0145] To a solution of (4S)-4-[(2-methylpropan-2-yl)oxycarbonylamino]-5-oxo-5-phenylmethoxypentanoic acid (Boc-Glu-OBn, CAS number 30924-93-7) (200 g, 592.82 mmol), N-hydroxyphthalimide (106 g, 649.78 mmol, 1.10 equivalents), and DMAP (3.6 g, 29.47 mmol, 0.05 equivalents) in THF (2 L) was added dropwise DIC (138 mL, 1.54 equivalents) at 0°C under a nitrogen atmosphere. The reaction mixture was stirred at 25°C for 16 hours, the solids were removed by filtration, and the filtrate was evaporated under reduced pressure. The residue was diluted with toluene, the resulting solids were removed by filtration, and the filtrate was evaporated under reduced pressure. The residue was purified by recrystallization (acetone / heptane) to give compound 12a (1-O-benzyl 5-O-(1,3-dioxoisoindol-2-yl)(2S)-2-[(2-methylpropan-2-yl)oxycarbonylamino]pentanedioate) (230 g, 80%). LCMS (ESI) m / z = 505.2 (M+Na) + retention time: 0.992 min (analysis condition SMDmethod_16).

[0146] Nickel bromide trihydrate (NiBr ２ ・3H ２ O) (4 g, 0.07 equivalents) and 4,4'-di-tert-butyl-2,2'-bipyridyl (dtbbpy, CAS number 72914-19-3) (3.9 g, 14.55 mmol, 0.07 equivalents) were added to DMA (500 mL), and the mixture was stirred at 50°C for 2 hours under a nitrogen atmosphere to prepare a nickel solution.

[0147] The nickel solution prepared above was added to a mixture of compound 12a (100 g, 207.3 mmol), zinc powder (70 g, 5 equivalents), and 4-bromo-2-chloro-1-(trifluoromethyl)benzene (CAS No. 467435-07-0, 160 g, 617 mmol, 3 equivalents) in DMA (500 mL), and the mixture was stirred at 25°C for 16 hours. To the reaction mixture was added an aqueous solution of EDTA.2Na (10%), followed by extraction with ethyl acetate. The combined organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, and the solvent was distilled off under reduced pressure. The resulting residue was purified by silica gel column chromatography (ethyl acetate / petroleum ether) to give compound 12b (75 g, 77%). LCMS (ESI) m / z = 494 (M + Na) + retention time: 2.863 minutes (analysis condition: SMD method_17).

[0148] A toluene solution (900 mL) of compound 12b (75 g, 158.93 mmol) was cooled to 0°C, and trifluoromethanesulfonic acid (TfOH) (42 mL, 3.00 equivalents) was added dropwise. After stirring at room temperature for 1 hour, water (75 mL) was added. This mixture was extracted with water, and the combined aqueous layer was extracted with ethyl acetate. The combined organic layer was washed with water, dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure. Acetonitrile / water (900 / 900 mL) was added to the residue, and the pH was adjusted to 7 with aqueous sodium hydroxide (48%). Fmoc-OSu (51.2 g, 151.93 mmol, 0.95 equivalents) was added to this solution, and the mixture was stirred at room temperature for 16 hours while maintaining the pH at 7.8 to 8.0. The reaction mixture was filtered, and the filtrate was adjusted to pH 2 with 6 N hydrochloric acid. The precipitated solid was collected by filtration and dried at 50°C to obtain compound 12 (Fmoc-Hph(4-CF ３ -3-Cl)-OH) was obtained (70 g, 87%). LCMS (ESI) m / z=526 (M+Na)+ Retention time: 2.180 minutes (Analysis conditions SMDmethod_21)

[0149]

[0150] Compound 11 was used to prepare Fmoc-Phe(4-CF ３ )-OH, Fmoc-MeGly-OH, compound 12 (Fmoc-Hph(4-CF ３The same peptide elongation reaction as in Example 1 was carried out using Fmoc-Pro-OH and Fmoc-cLeu-OH, to give resin-supported compound 9a.

[0151] The resin-supported compound 9a (4.2 g) and DCM (45 mL) were added to a reaction vessel equipped with a filter to swell the resin. After removing the DCM from the reaction vessel, the resin was washed twice with DMF (30 mL). A DBU solution in DMF (2% v / v, 30 mL) was added and the mixture was shaken at room temperature for 10 minutes, after which the reaction solution was removed from the reaction vessel. The resin remaining in the reaction vessel was washed four times with NMP (30 mL), washed with a DCM solution (30 mL) of triethylamine hydrochloride (520 mg, 3.78 mmol), washed four times with DCM (30 mL), and washed four times with THF (30 mL) to obtain resin-supported compound 9b.

[0152] To the reaction vessel containing the resin-supported compound 9b, a THF solution (27.5 mL) of nosyl chloride (1.675 g, 7.56 mmol) and 2,4,6-collidine (2.5 mL, 18.9 mmol) were added, and the reaction vessel was shaken at room temperature for 2 hours. The reaction solution was then removed, and the resin was washed four times with THF (30 mL) and four times with DCM (30 mL) to obtain resin-supported compound 9c.

[0153] Two grams of the resulting resin-supported compound 9c was added to DCM (20 mL) to swell the mixture. After removing the DCM, the mixture was washed four times with THF (14 mL). A THF solution (7 mL) of triphenylphosphine (1.18 g, 4.50 mmol) and a THF solution (7 mL) of DIAD (0.876 mL, 4.50 mmol) were mixed and allowed to stand for 15 minutes. 3-Buten-1-ol (0.773 mL, 9.00 mmol) was then added and allowed to stand for 5 minutes. This mixture was added to the resin and shaken at room temperature for 45 minutes, after which the reaction solution was removed. The resin was washed five times with THF (14 mL) and five times with DCM (14 mL) to obtain resin-supported compound 9d.

[0154] DCM (20 mL) was added to resin-supported compound 9d to swell it. After removing the DCM, the resin was washed four times with THF (14 mL). A solution of DBU (0.678 mL, 4.50 mmol) in NMP (7 mL) and a solution of 1-dodecanethiol (2.05 mL, 0.262 mmol) in NMP (7 mL) were added and the mixture was shaken at 40°C for 1 hour. The reaction solution was then removed, and the resin was washed five times with DMF (14 mL) and five times with DCM (14 mL) to obtain resin-supported compound 9e.

[0155] Using the obtained resin-supported 9e, the same peptide elongation reaction as in Example 1 was carried out using Fmoc-MeLeu-OH, Fmoc-Ile-OH, Fmoc-MeGly-OH, and Fmoc-MeAlgly-OH, followed by cleavage of the elongated peptide from the resin, cyclization of the cleaved peptide (using (1-cyano-2-ethoxy-2-oxoethylideneaminooxy)dimethylamino-morpholino-carbenium hexafluorophosphate (COMU) as the cyclization reagent), and purification of the cyclic peptide compound, to obtain compound 9f.

[0156] Compound 9f (78 mg, 0.050 mmol) and Stewart-Grubbs catalyst (7.1 mg, 0.012 mmol) were dissolved in DCE (10 mL), degassed, and stirred at 50°C under a nitrogen atmosphere for 6 hours. Stewart-Grubbs catalyst (7.1 mg, 0.012 mmol) was added, degassed, and stirred at 50°C under a nitrogen atmosphere for an additional 15 hours. The solvent was then distilled off under reduced pressure. This was purified by reverse-phase chromatography (acetonitrile containing 0.1% formic acid / distilled water containing 0.1% formic acid) to give compound 9g (18 mg, 23%).

[0157] Compound 9g (16 mg, 0.010 mmol) was dissolved in ethyl acetate (1 mL), platinum(IV) oxide (1.6 mg, 0.007 mmol) was added, and the mixture was stirred at room temperature under a hydrogen atmosphere for 4 hours. The platinum(IV) oxide was removed by filtration, and the resulting filtrate was evaporated under reduced pressure. The resulting crude product was purified by reverse-phase chromatography (0.1% formic acid in acetonitrile / 0.1% formic acid in distilled water) to obtain compound 9 (10 mg, 65%).

[0158] Synthesis of Compound 10

[0159] Compound 11c, ((2S)-2-[9H-fluoren-9-ylmethoxycarbonyl(methyl)amino]-4-oxo-4-prop-2-enoxybutanoic acid) (1.5 g, 3.66 mmol) was used as the starting material, and compound 13b (1.47 g, 92%) was obtained in the same manner as in the synthesis of compound 11d, except that a THF solution of dimethylamine (2 mol / L) was used instead of morpholine. LCMS (ESI) m / z = 437 (M+H) + retention time: 0.88 min (analysis condition SQDFA05).

[0160] Compound 13c (5.0 g, 43%) was obtained using compound 13b as a starting material in the same manner as in the synthesis of compound 11e. LCMS (ESI) m / z = 576.5 (M + H) + Retention time: 1.02 min (Analysis condition SQDFA05).

[0161] Using the obtained compound 13c, compound 13d (0.786 g, 92%) was obtained as a resin-supported compound under the same reaction conditions as those used to synthesize compound 11f. LCMS (ESI) m / z = 536 (M + H) + Retention time: 0.85 min (Analysis condition SQDFA05).

[0162] Using the obtained compound 13d, compound 13 (2.72 g) was obtained under the same reaction conditions as in the synthesis of compound 11. The loading amount was calculated to be 0.345 mmol / g in the same manner as in the synthesis of compound 11.

[0163]

[0164] Nickel bromide trihydrate (NiBr ２ ・3H ２ O) (13.5 g, 49.7 mmol, 0.3 equivalents) and 4,4'-di-tert-butyl-2,2'-bipyridyl (dtbbpy, CAS number 72914-19-3) (13.3 g, 49.7 mmol, 0.3 equivalents) were added to DMA (400 mL), and the mixture was stirred at 50°C for 3 hours under a nitrogen atmosphere to prepare a nickel solution.

[0165] A mixture of compound 12a (1-O-benzyl 5-O-(1,3-dioxoisoindol-2-yl)(2S)-2-[(2-methylpropan-2-yl)oxycarbonylamino]pentanedioate) (80 g, 166 mmol), zinc powder (54.2 g, 829 mmol, 5 equivalents), and 4-bromo-1,3-difluoro-2-(trifluoromethyl)benzene (CAS No. 156243-64-0, 86.6 g, 332 mmol, 2 equivalents) in DMA (400 mL) was stirred at room temperature for 1 hour under a nitrogen atmosphere, and the previously prepared nickel solution was added, followed by stirring at room temperature for 16 hours. To the reaction mixture was added an aqueous solution of EDTA.2Na (800 mL, 10%), and the solid was removed by filtration. The filtrate was extracted with ethyl acetate, and the combined organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure. The resulting residue was purified by silica gel column chromatography (ethyl acetate / petroleum ether) to give Compound 14a (57.2 g, 69%). LCMS (ESI) m / z = 496 (M + Na) + Retention time: 1.544 min (Analysis condition SMDmethod_15).

[0166] A toluene mixture (690 mL) of compound 14a (57.2 g, 121 mmol) was cooled to 0°C, and trifluoromethanesulfonic acid (TfOH) (54.4 g, 362 mmol, 3 equivalents) was added dropwise. After stirring at room temperature for 1 hour, water (58 mL) was added. This mixture was extracted with water, and the combined aqueous layer was extracted with ethyl acetate. The combined organic layers were washed with water and dried over anhydrous sodium sulfate. The solvent was then distilled off under reduced pressure to obtain 60 g of a residue. Acetonitrile / water (400 / 400 mL) was added to the residue, and the pH was adjusted to 7 with aqueous sodium hydroxide (48%). Fmoc-OSu (36.6 g, 108.6 mmol, 0.9 equivalents) was added to this solution, and the pH was adjusted to 8.0 with aqueous sodium hydroxide (48%). The mixture was then stirred at room temperature for 16 hours. The reaction mixture was filtered while washing with acetonitrile / water (1 / 1) to remove solid components. The filtrate was diluted with acetonitrile and acidified with 6N hydrochloric acid. The precipitated solid was collected by filtration to give Compound 14 (52 g, 83%). LCMS (ESI) m / z = 528 (M + Na) + Retention time: 3.538 minutes (Analysis condition: SMDmethod_14).

[0167]

[0168] To a solution of compound 15a (N-([(9H-fluoren-9-yl)methoxy]carbonyl)-4-(trifluoromethyl)-L-phenylalanine) (10.0 g, 22.0 mmol) in dichloromethane (200 mL) were added magnesium sulfate (6.61 g, 54.9 mmol) and paraformaldehyde (1.98 g, 65.9 mmol) at room temperature, followed by dropwise addition of boron trifluoride diethyl ether complex (2.78 mL, 22.0 mmol). The reaction mixture was stirred at room temperature for 1 hour, filtered through silica gel (24 g), and then concentrated under reduced pressure to give crude product 15b (10.5 g, quant.). LCMS (ESI) m / z = 468 (M+H) + retention time: 1.45 min (analysis conditions SMD method_04).

[0169] The resulting crude product 15b (10.3 g, 22.0 mmol) was dissolved in dichloromethane (200 mL). Allyltrimethylsilane (12.2 mL, 77.0 mmol) was added under a nitrogen atmosphere, followed by the dropwise addition of boron trifluoride diethyl ether complex (7.0 mL, 54.9 mmol). The reaction mixture was stirred at room temperature for 6 hours, after which allyltrimethylsilane (12.2 mL, 77.0 mmol) and boron trifluoride diethyl ether complex (7.0 mL, 54.9 mmol) were added, and the mixture was stirred for 3 days. Water (40 mL) was added to the reaction mixture, which was stirred at room temperature for 10 minutes. The mixture was then filtered through Celite. The filtrate was concentrated under reduced pressure, and the resulting residue was dissolved in toluene (24 mL). The organic layer was washed with water (48 mL), then hexane (100 mL) was added, and the mixture was extracted with a mixture of 3.5% aqueous potassium bicarbonate and acetonitrile (2:1, 300 mL). Phosphoric acid was added to the resulting aqueous layer until the pH reached 3, followed by extraction with ethyl acetate (200 mL). The organic layer was washed with saturated brine (100 mL), dried over anhydrous magnesium sulfate, and then filtered. The resulting solution was concentrated under reduced pressure to give compound 15c (10.3 g, 92%). LCMS (ESI) m / z = 510 (M + H) + retention time: 1.00 min (analysis condition SMD method_08).

[0170] To a solution of compound 15c (67.5 g, 132 mmol) and sarcosine tert-butyl ester hydrochloride (25.3 g, 139 mmol) in NMP (379 mL) was added HATU (60.4 g, 159 mmol), followed by dropwise addition of DIPEA (69.2 mL, 397 mmol). The reaction mixture was stirred at room temperature for 1.5 hours, after which toluene (1.0 L, 15 v / w) was added, and the organic layer was washed with water (473 mL, 7 v / w), 3.5% aqueous potassium bicarbonate (473 mL, 7 v / w), and 1 M aqueous potassium dihydrogen phosphate (473 mL, 7 v / w). The organic layer was dried through an anhydrous magnesium sulfate pad and washed with toluene (67.5 mL, 1 v / w). The resulting filtrate and washings were combined to give compound 15d, which was used in the subsequent reaction as a solution. LCMS (ESI) m / z=637 (M+H)+ Retention time: 1.66 minutes (Analysis conditions SMD method_10)

[0171] DBU (1.97 mL, 13.2 mmol) was added to a toluene solution of compound 15d (132 mmol) at room temperature. The reaction mixture was stirred at room temperature for 17 hours and then washed with 1 M aqueous potassium dihydrogen phosphate solution (420 mL, 5 v / w). The resulting organic layer was diluted with n-hexane (840 mL, 10 v / w) and extracted twice with a mixture of 2 M hydrochloric acid (99 mL, 198 mmol), water (840 mL, 10 v / w), and acetonitrile (588 mL, 7 v / w). The resulting aqueous layer was adjusted to pH 9.5 with 3.3 M aqueous potassium phosphate solution and then extracted with ethyl acetate (840 mL, 10 v / w). The organic layer was concentrated under reduced pressure, and the resulting residue was dissolved in ethyl acetate (840 mL, 10 v / w), washed with brine (420 mL, 5 v / w), dried over anhydrous magnesium sulfate, and filtered. The resulting solution was concentrated under reduced pressure to give compound 15e (49.0 g, 90%). LCMS (ESI) m / z = 415 (M+H) + Retention time: 1.31 minutes (Analysis conditions SMD method_10)

[0172] DIPEA (72.3 mL, 414 mmol) was added to a solution of compound 15e (49.0 g, 118 mmol) in dichloromethane (276 mL). The resulting mixture was added dropwise over 30 minutes to a separately prepared solution of (S)-(1-chloro-1-oxopent-4-en-2-yl)(methyl)carbamate (9H-fluoren-9-yl)methyl (described below) in dichloromethane (276 mL). The resulting reaction mixture was stirred at room temperature for 17 hours, after which the organic layer was washed with a 5% aqueous solution of sodium dihydrogen phosphate (500 mL, 10 v / w), dried over anhydrous magnesium sulfate, filtered, and the solution was concentrated under reduced pressure. The resulting crude product was purified by normal-phase silica gel chromatography (n-hexane-ethyl acetate) to give compound 15f (45.0 g, 51%).

[0173] (S)-(1-chloro-1-oxopent-4-en-2-yl)(methyl)carbamate (9H-fluoren-9-yl)methyl was prepared as follows. To a solution of (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)(methyl)amino)pent-4-enoic acid (41.6 g, 118 mmol) in dichloromethane (296 mL) was added thionyl chloride (21.6 mL, 296 mmol), followed by dropwise addition of DMF (0.92 mL, 11.8 mmol). The resulting reaction mixture was stirred at room temperature for 15 minutes and then concentrated under reduced pressure. Toluene (20 mL, 5 v / w) was added to the resulting residue, followed by concentration under reduced pressure. The resulting crude product was dissolved in dichloromethane (276 mL) and used in the above reaction. LCMS (ESI) m / z=748 (M+H)+ Retention time: 1.71 minutes (Analysis conditions SMD method_10)

[0174] A solution of compound 15f (16.7 g, 22.3 mmol), benzoquinone (0.24 g, 2.23 mmol), and STEWART-GRUBBS CATALYST (0.51 g, 0.89 mmol) in dichloroethane (891 mL) was degassed five times and stirred under a nitrogen atmosphere at 80°C for 4 hours. The reaction mixture was cooled to room temperature and then concentrated under reduced pressure. The resulting crude product was purified by normal phase silica gel column chromatography (hexane-ethyl acetate) to give compound 15g (33.9 g, 39%). LCMS (ESI) m / z = 720 (M+H) + retention time: 1.00 min (analysis conditions: SMD method_07).

[0175] Under a nitrogen atmosphere, hexamethyldisilazane (24.7 mL, 118 mmol) was added to a solution of 15 g (33.9 g, 47.1 mmol) in ethyl acetate (236 mL), and then trimethylsilyl trifluoromethanesulfonate (17.0 mL, 94 mmol) was added dropwise while cooling in a water bath (25 °C). The reaction mixture was stirred at room temperature for 1.5 hours, and then 5% aqueous disodium hydrogen phosphate solution (340 mL, 10 v / w) was added while cooling in an ice bath to quench the reaction. Water (170 mL, 5 v / w) and phosphoric acid were added to the resulting mixture to adjust the aqueous layer to pH 3. The separated organic layer was washed with saturated brine (340 mL, 10 v / w), dried over anhydrous magnesium sulfate, filtered, and the solution was concentrated under reduced pressure. The resulting crude product was purified by reverse-phase column chromatography (acetonitrile containing 0.1% formic acid / distilled water containing 0.1% formic acid), and the collected fractions were purified by reverse-phase column chromatography (acetonitrile / distilled water) to obtain Compound 15 (22.4 g, 72%). LCMS (ESI) m / z = 664 (M+H) + Retention time: 0.63 min (Analysis condition: SMD method_06).

[0176] Compound 13 was used to synthesize Fmoc-MeAbu-OH, Fmoc-Ile-OH, Fmoc-MeGly-OH, Compound 15, and Compound 14 (Fmoc-Hph(4-CF ３Compound 10 was obtained by carrying out the same peptide elongation reaction as in Example 1 using Fmoc-Hyp(Et)-OH, Fmoc-cVal-OH, cleavage of the elongated peptide from the resin, cyclization of the cleaved peptide (using (1-cyano-2-ethoxy-2-oxoethylideneaminooxy)dimethylamino-morpholino-carbenium hexafluorophosphate (COMU) as the cyclization reagent), and purification of the cyclic peptide compound.

[0177] Amino acid abbreviations are listed in Table 1. In this specification, when -OH is listed in addition to an amino acid abbreviation in Table 1, it means that the C-terminus of that amino acid is a carboxyl group. For example, Gly-OH means that the C-terminus of glycine is a carboxyl group, and Fmoc-Gly-OH means that the N-terminal amino group of glycine is protected with an Fmoc group and the C-terminus is a carboxyl group.

[0178]

[0179] The structural formulae of compounds 1 to 10 are shown in Tables 2 to 4.

[0180]

[0181] The LC / MS analysis conditions are shown in Table 5.

[0182] The mass spectrum values ​​and liquid chromatography retention times of the obtained compounds 1 to 10 are shown in Table 6.

[0183] The molecular weights of Compounds 1 to 10 are as follows: Compound 1: 1442.2 Compound 2: 1385.8 Compound 3: 1399.8 Compound 4: 1433.8 Compound 5: 1413.8 Compound 6: 1454.2 Compound 7: 1410.1 Compound 8: 1459.7 Compound 9: 1546.2 Compound 10: 1505.6

[0184] [Example 2] Effect of surfactant (Example 2-1) In vitro solubility evaluation The solubility of compounds 1 and 2 was evaluated. 50 mM phosphate buffer (PPB, pH 6.5) was added to the lyophilized compound powder, and the mixture was shaken (room temperature, 1800 rpm, 24 hours), filtered, and the compound concentration of the filtrate was measured by LC / MS / MS. The solubility of compounds 1 and 2 in the presence of surfactant was evaluated. Cremophor EL (CreEL) (Sigma-Aldrich, "Kolliphor EL"; generic name: polyoxyethylene castor oil, average number of moles of ethylene oxide added: 35), Vitamin E TPGS (Sigma-Aldrich), an aqueous solution containing 1% TW80 (Nacalai Tesque), an aqueous solution containing 1.5% Ploxamer 188 (BASF), or an aqueous solution containing 0.5% Triton X-100 were added to the compound powder. The mixture was stirred with a stirrer (room temperature, 20 hours), filtered, and the compound concentration of the filtrate was measured by LC / UV. Solubility (μg / mL) was calculated from the measured compound concentration. The results are shown in Table 7. Compounds 1 and 2 were confirmed to have greater solubility in the presence of surfactants than in the absence of surfactants. In particular, greater solubility was observed in the presence of CreEL or VitaminE TPGS than in the presence of other surfactants (Table 7). This confirms that the use of CreEL or VitaminE TPGS shows excellent solubility improvement independent of the compound, compared to when no surfactant is used or when other surfactants are used. Based on the above, CreEL and VitaminE TPGS were selected as substrates that improve solubility independent of the compound, and oral absorbability was evaluated.

[0185]

[0186] (Example 2-2) Preparation of Formulations Example formulations (1) and (2) were prepared by mixing and stirring compounds 1 and 2, water for injection (Otsuka Pharmaceutical Factory, Inc.), dimethyl sulfoxide (DMSO) (Wako Pure Chemical Industries, Ltd.), and Cremophor EL to obtain the compositions shown in Table 8. Compounds 1 and 2 were dissolved in dimethyl sulfoxide to a concentration of 3 mg / mL before mixing.

[0187] Comparative preparations (1-1) to (2-2) were prepared by mixing and stirring Compounds 1 and 2, water for injection (Otsuka Pharmaceutical Factory, Inc.), dimethyl sulfoxide (Wako Pure Chemical Industries, Ltd.), and Vitamin E TPGS (Sigma-Aldrich) to the compositions shown in Table 8. Compounds 1 and 2 were dissolved in dimethyl sulfoxide to a concentration of 3 mg / mL before mixing.

[0188]

[0189] (Example 2-3) Rat PK Study (3 mg / kg) The blood kinetics of Example Formulations (1) and (2) and Comparative Formulations (1-1), (1-2), (2-1), and (2-2) after oral administration to rats was evaluated. Male rats (WIST, 7 weeks old, manufactured by Japan Institute of Medical Science Animal Materials: 4 rats per group) were orally administered either Example Formulations (1) and (2) or Comparative Formulations (1-1), (1-2), (2-1), or (2-2) at a dose of 3 mg / kg of Compound 1, and blood samples were collected over time from the jugular vein using a syringe treated with heparin as an anticoagulant up to 24 hours after administration. Plasma was separated from the blood by centrifugation, and after deproteinization with acetonitrile, the plasma concentrations of Compounds 1 and 2 were measured using an LC / MS / MS system. From the obtained plasma concentration time courses, pharmacokinetic parameters were calculated by non-compartmental analysis using the analytical software Phoenix WinNonlin 8.2 (Certara L.P.). The results are shown in Table 9.

[0190] Pharmacokinetic parameters were calculated as the area under the plasma drug concentration-time curve (AUClast; ng h / mL) from time 0 after administration to the final time point at which a concentration exceeding the limit of quantitation was obtained, and the maximum plasma concentration after administration (Cmax; ng / mL). For Compounds 1 and 2, the AUClast values ​​for the groups administered with Example Formulations (1) and (2) were all greater than the AUClast values ​​for the groups administered with Comparative Formulations (1-1), (1-2), (2-1), and (2-2), and an increase in Cmax was also observed (Table 9). These results demonstrate that the use of Cremophor EL results in higher absorbability of low-membrane-permeable compounds compared to when Cremophor EL is not used.

[0191]

[0192] [Example 3] Evaluation of physical properties of cyclic peptide compounds (Example 3-1) Evaluation of Caco-2 membrane permeability Caco-2 cells were cultured on a 96-well transwell for 3 weeks, after which DMEM + FaSSIF (1% DMSO) + compound was added to the apical side and DMEM was added to the basal side. ２ Pre-incubation was carried out for 18 to 24 hours at 37°C with shaking at 80 rpm. After pre-incubation, the pre-incubation solution on the apical and basal sides was removed by aspiration and washed. FaSSIF / HBSS buffer (1% DMSO) (pH 6.0) containing the compound was added to the apical side, and HBSS buffer (4% BSA) (pH 7.4) was added to the basal side, and measurement of membrane permeability was then initiated. Each well was incubated at 5% CO ２ The tubes were shaken at 37°C and 80 rpm, and 180 minutes after the start of shaking, a sample was taken from the basal side, and the permeation amount was measured by LC / MS. The membrane permeability coefficient (Papp) was calculated from the permeation amount. Note that the term "Caco-2 (cm / sec)" in the columns of the tables in this specification is used synonymously with "Papp (cm / sec)."

[0193] Caco-2 values ​​of cyclic peptide compounds

[0194] (Example 3-2) Evaluation of Lipid Solubility of Cyclic Peptide Compounds The ClogP values ​​of Compounds 1 to 10 are as follows.

[0195] (Example 3-3) Evaluation of Solubility of Cyclic Peptide Compounds The solubility of cyclic peptide compounds was evaluated. 50 mM phosphate buffer (PPB, pH 6.5) was added to the lyophilized compound powder, and the mixture was shaken (37°C, 1800 rpm, 22 to 24 hours). After filtration, the compound concentration of the filtrate was measured by LC / MS / MS. The solubility was calculated from the measured compound concentration. The solubilities of compounds 3, 4, 6, 7, and 8 were 11 μg / mL, 1 μg / mL, 1 μg / mL, 15 μg / mL, and 5 μg / mL, respectively. These results confirmed that the cyclic peptide compounds exhibited uniformly low solubility.

[0196] The solubility of cyclic peptide compounds in the presence of surfactants was evaluated. Aqueous solutions containing 0.3% or 1% Cremophor EL (CreEL) were added to lyophilized compound powders, followed by shaking (25°C, 1000 rpm, 22-24 hours), filtration, and measurement of the compound concentration in the filtrate by LC / UV. Solubility (μg / mL) was calculated from the measured compound concentration. The results are shown in Table 12. All cyclic peptide compounds exhibited a solubility of 100 μg / mL or more in aqueous CreEL solutions. These results confirmed that the use of CreEL improved the solubility of low-membrane-permeable compounds compared to when no surfactant was used.

[0197]

[0198] Example 4 Preparation of Formulations Example 4-1 Preparation of Formulations Using Cremophor EL The cyclic peptide compound, water for injection (manufactured by Shanghai Pujin Lin Pharmaceutical Co., Ltd. or Otsuka Pharmaceutical Factory, Inc.), dimethyl sulfoxide (manufactured by Wako Pure Chemical Industries, Ltd.), and Cremophor EL (manufactured by Sigma-Aldrich, "Kolliphor EL", generic name: polyoxyethylene castor oil, average number of moles of ethylene oxide added: 35) were mixed and stirred to obtain the compositions shown in Tables 13 to 33 to prepare example formulations and comparative formulations.

[0199]

[0200]

[0201]

[0202]

[0203]

[0204]

[0205]

[0206]

[0207]

[0208]

[0209]

[0210]

[0211]

[0212]

[0213]

[0214]

[0215]

[0216]

[0217]

[0218]

[0219]

[0220] [Example 5] PK Study (Example 5-1) Mouse PK Study (30 mg / kg) The blood kinetics of the Example Formulation and the Comparative Formulation after oral administration in mice were evaluated. Male mice (C57BL / 6J, 7-11 weeks old, manufactured by SiBeiFu Laboratory Animal Technology Co. Ltd. or Charles River Japan, Inc.; 3 mice per group) were orally administered one of the prepared Example Formulations or Comparative Formulations at a dose of 30 mg / kg of Compounds 3-10. Blood samples were collected over time from the jugular vein and posterior vena cava, or from the dorsal pedicle vein and heart using heparin as an anticoagulant up to 24 hours after administration. Plasma was separated from the blood by centrifugation, and the plasma concentrations of Compounds 3-10 were measured using an LC / MS / MS system after deproteinization with acetonitrile or liquid-liquid extraction with ethyl acetate. From the obtained plasma concentration time courses, pharmacokinetic parameters were calculated by non-compartmental analysis using the analytical software Phoenix WinNonlin 8.2 (Certara LP). The results are shown in Tables 34 to 42.

[0221] Pharmacokinetic parameters were calculated as the area under the plasma drug concentration-time curve (AUClast; ng h / mL) from 0 hours after administration to the final time point at which a concentration exceeding the limit of quantitation was obtained, and the maximum plasma concentration after administration (Cmax; ng / mL). The AUClast ratio was calculated as the ratio (comparator AUC) of the example formulation or comparative formulation to the AUClast (maximum AUC) of the example formulation that showed the maximum AUClast upon administration of the same compound within the same test (comparator AUC / maximum AUC). In the cyclic peptide compound group, it was confirmed that the AUClast of the example formulation administration group was greater than the AUClast of the comparative formulation administration group (AUClast ratio of 0.8 or more), and an increase in Cmax was also observed. As shown in Figure 1, it was confirmed that for all compounds, by using a Cremophor EL formulation in which the Cremophor EL content based on 100% by volume of the formulation was 2.0% to 7.5% by volume, the low membrane permeability compounds exhibited higher absorbability (AUClast ratio of 0.8 or greater) compared to the cases in which a Cremophor EL content based on 100% by volume of the formulation was less than 2.0% by volume or more than 7.5% by volume.

[0222]

[0223]

[0224]

[0225]

[0226]

[0227]

[0228]

[0229]

[0230]

[0231] Table 43 shows the percentage (%) of plots with an AUClast ratio of 0.8 or more relative to the total number of plots for each Cremophor EL concentration.

[0232] As shown in Table 43, when the Cremophor EL concentration was greater than 7.5% by volume, the probability of an AUClast ratio of 0.8 or greater was 14%, and when the Cremophor EL concentration was less than 2% by volume, the probability of an AUClast ratio of 0.8 or greater was 0%, whereas when the Cremophor EL concentration was 2.0 to 7.5% by volume, the percentage of AUClast ratios of 0.8 or greater improved to 100%. In other words, it was confirmed that by using 2.0 to 7.5% by volume of Cremophor EL in a cyclic peptide compound, a compound with low membrane permeability exhibited higher absorbability (AUClast ratio of 0.8 or greater) compared to when Cremophor EL concentrations of less than 2.0% by volume and greater than 7.5% by volume were used.

[0233] (Example 5-2) Mouse PK study (3 mg / kg) The blood kinetics of the Example formulation and the Comparative formulation after oral administration in mice were evaluated. Male mice (C57BL / 6, 10 weeks old, manufactured by SiBeiFu Laboratory Animal Technology Co. Ltd. or Charles River Japan, Inc.; 3 mice per group) were orally administered one of the prepared Example formulations or Comparative formulations at a dose of 3 mg / kg of Compounds 3, 4, and 6-10. Blood samples were collected over time from the jugular vein and posterior vena cava, or from the dorsal pedicle vein and heart using heparin as an anticoagulant up to 24 hours after administration. Plasma was separated from the blood by centrifugation, and the plasma concentrations of Compounds 3, 4, and 6-10 were measured using an LC / MS / MS system after deproteinization with acetonitrile or liquid-liquid extraction with ethyl acetate. From the obtained plasma concentration time courses, pharmacokinetic parameters were calculated by non-compartmental analysis using the analytical software Phoenix WinNonlin 8.2 (Certara LP). The results are shown in Tables 44 to 56.

[0234] Pharmacokinetic parameters were calculated as the area under the plasma drug concentration-time curve (AUClast; ng h / mL) from 0 hours after administration to the final time point at which a concentration exceeding the limit of quantitation was obtained, and the maximum plasma concentration after administration (Cmax; ng / mL). The AUClast ratio was calculated as the ratio (comparator AUC) of the example formulation or comparative formulation to the AUClast (maximum AUC) of the example formulation that showed the maximum AUClast when the same compound was administered in the same test (comparator AUC / maximum AUC). In the cyclic peptide compound group, it was confirmed that the AUClast of the example formulation administration group was more likely to be greater than the AUClast of the comparative formulation administration group (AUClast ratio of 0.7 or more), and an increase in Cmax was also observed. As shown in Figure 2, for all compounds, it was confirmed that the use of 2.0 vol% to 7.5 vol% Cremophor EL resulted in high absorbability (AUClast ratio of 0.7 or greater) for compounds with low membrane permeability compared to the use of Cremophor EL concentrations less than 2.0 vol% and greater than 7.5 vol%.

[0235]

[0236]

[0237]

[0238]

[0239]

[0240]

[0241]

[0242]

[0243]

[0244]

[0245]

[0246]

[0247]

[0248] Table 57 shows the percentage (%) of plots with an AUClast ratio of 0.7 or more relative to the total number of plots for each Cremophor EL concentration.

[0249] As shown in Table 57, at a dose of 3 mg / kg, for example, when the Cremophor EL concentration was greater than 7.5 vol%, the probability of an AUClast ratio of 0.7 or greater was 0%, while when the Cremophor EL concentration was less than 2.0 vol%, the probability of an AUClast ratio of 0.7 or greater was 42%. However, when the Cremophor EL concentration was 2.0 to 7.5 vol%, the proportion of cases with an AUClast ratio of 0.7 or greater improved to 86%. In other words, it was confirmed that the use of 2.0 to 7.5 vol% Cremophor EL in a cyclic peptide compound exhibited higher absorbability (AUClast ratio of 0.7 or greater) compared to when Cremophor EL concentrations of less than 2.0 vol% and greater than 7.5 vol% were used.

[0250] Based on the above results, screening for oral absorption at a specific Cremophor EL concentration can achieve high oral absorption and high plasma concentrations (AUC and Cmax). In other words, it is possible to increase the oral absorption of compounds with poor oral absorption, a barrier to drug discovery, according to their potential. This reduces the dropout rate due to false negatives and allows for screening of compounds with excellent oral absorption, enabling the selection of optimal compounds. Furthermore, achieving high oral absorption and high plasma concentrations allows for precise evaluation of various compound characteristics essential for drug development, such as oral bioavailability, pharmacological activity upon oral administration, safety upon oral administration, pharmacokinetics upon oral administration, and compound physical properties, enabling better drug development decisions.

Claims

1. A method for screening candidate compounds using a formulation, The method, wherein the formulation contains the test compound and an oily component containing a polyoxyethylene structure, and the content of the oily component in 100% by volume of the formulation is 2.0% by volume or more and 7.5% by volume or less.

2. The method according to claim 1, wherein the oily component containing the polyoxyethylene structure is at least one selected from the group consisting of polyoxyethylene castor oil, polyoxyethylene hydrogenated castor oil, and polyoxyethylene sorbitan fatty acid ester.

3. The method according to claim 1, wherein the oily component containing the polyoxyethylene structure is polyoxyethylene castor oil.

4. The method according to any one of claims 1 to 3, wherein the test compound is a peptide compound.

5. The method according to claim 4, wherein the test compound is a peptide compound containing 5 to 30 amino acid residues.

6. The method according to claim 4, wherein the peptide compound is a peptide compound having a cyclic portion.

7. The method according to any one of claims 1 to 3, wherein the preparation is an oral preparation.

8. The method according to any one of claims 1 to 3, comprising administering the preparation to the target.

9. The method according to any one of claims 1 to 3, comprising selecting the candidate compound using at least one selected from the group consisting of (1), (2), and (3) below as an indicator; (1) Medicinal effects, (2) toxicity; (3) Pharmacokinetic properties.

10. The method according to any one of claims 1 to 3, wherein a candidate compound is selected from a plurality of test compounds.

11. The method according to claim 8, wherein the subject is a non-human animal.

12. The method according to claim 8, wherein the dose of the test compound administered is 0.1 mg / kg or more and 1000 mg / kg or less.