JP2006265107A - Therapeutic agent for glaucoma - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a therapeutic and/or prophylactic agent for glaucoma containing thiadiazoline derivative or its pharmacologically acceptable salt as an active ingredient. <P>SOLUTION: The present invention provides a therapeutic and/or prophylactic agent for glaucoma containing a thiadiazoline derivative represented by general formula (I) [wherein n is an integer of 1-3; R<SP>1</SP>is a hydrogen atom and R<SP>2</SP>is a lower alkyl or R<SP>1</SP>and R<SP>2</SP>together form an alkylene; R<SP>3</SP>is a lower alkyl; R<SP>4</SP>is NHSO<SB>2</SB>R<SP>6</SP>(wherein R<SP>6</SP>is hydroxy or the like); R<SP>5</SP>is an aryl or the like] or its pharmacologically acceptable salt. <P>COPYRIGHT: (C)2007,JPO&INPIT

Description

The present invention relates to a therapeutic and / or prophylactic agent for glaucoma containing a thiadiazoline derivative or a pharmacologically acceptable salt thereof as an active ingredient.

  Glaucoma is a disease in which the optic nerve is damaged and the visual field becomes narrow, and an increase in intraocular pressure is said to be one of the etiologies. Glaucoma is a general term for diseases such as primary closed-angle glaucoma, primary open-angle glaucoma, and normal-tension glaucoma. It is one of the top diseases causing blindness, and there is a need for therapeutic drugs. Currently, reducing intraocular pressure is used as the main treatment. Prostaglandin derivatives, adrenaline α stimulants, adrenaline β blockers, and the like have been used as drugs for reducing intraocular pressure (Drugs, 59, p. 411 (2000)). Recently, Rho-associated protein kinase (ROCK) inhibitors have been reported to reduce intraocular pressure (Invest. Ophthalmol. Vis. Sci.), 42, p.137 (2001)). ROCK inhibitors are thought to increase intraperitoneal permeability, which is the exit of aqueous humor, and reduce intraocular pressure. It has also been reported that the development of glaucoma involves the proliferation of vascular endothelial cells (Clin. Experimental Ophthalmol., 31, p. 492 (2003)). Ophthalmologica, 218, p. 344 (2004)). From these facts, it is considered that the outflow of aqueous humor can be promoted, intraocular pressure can be reduced, and glaucoma can be treated by improving the trabecular permeability and suppressing abnormal growth of vascular endothelial cells.

On the other hand, M-phase kinesin is a protein involved in M-phase spindle regulation, and plays an essential role in the M-phase progression of the cell cycle. One of the M-phase kinesins, M-phase kinesin easy five (Eg5), is a homotetrameric bipolar molecule that crosslinks two microtubules in the same direction toward the + (plus) end. Move and slide between two anti-parallel microtubules and move the-(minus) ends of the microtubules apart to separate the spindle poles to form a bipolar spindle structure It is known to be involved [Cell, 83, p. 1159 (1995), Journal of Cell Biology, 150, p. 975 (2000), Experimental Medicine, Vol. 17, p. 439 (1999)]. Therefore, inhibitors of Eg5 are considered promising as therapeutic agents for diseases involving cell proliferation [WO2001 / 98278, WO2002 / 56880, WO2002 / 57244, Trends in Cell Biology. ), Volume 12, p. 585 (2002)]. Examples of Eg5 inhibitors include quinazolin-4-one derivatives (WO2001 / 30768, WO2003 / 039460, etc.), triphenylmethane derivatives (WO2002 / 56880), thiadiazoline derivatives (see Patent Documents 1 to 3), and the like. Yes.
International Publication No. 2004-092147 Pamphlet International Publication No. 2004-111023 Pamphlet International Publication No. 2004-111024

  An object of the present invention is to provide a therapeutic and / or prophylactic agent for glaucoma containing a thiadiazoline derivative or a pharmacologically acceptable salt thereof as an active ingredient.

The present invention relates to the following (1) to (12).
(1) General formula (I)

{Wherein n represents an integer of 1 to 3,
R ¹ represents a hydrogen atom,
R ² represents lower alkyl,
Or R ¹ and R ² together represent alkylene,
R ³ represents lower alkyl,
R ⁴ is a hydrogen atom,
NHSO ₂ R ⁶ (wherein R ⁶ is a lower alkyl optionally having 1 to 2 substituents selected from the group consisting of hydroxy, lower alkoxy, amino, lower alkylamino and di-lower alkylamino, Or lower alkenyl)
NHR ⁷ wherein R ⁷ is a lower alkyl optionally having 1 or 2 substituents selected from the group consisting of hydroxy, lower alkoxy, amino, lower alkylamino and di-lower alkylamino, COR ⁸ Wherein R ⁸ is selected from the group consisting of hydroxy, lower alkoxy, amino, lower alkylamino, di-lower alkylamino, carboxy, phenyl, hydroxyphenyl, imidazolyl, guanidyl, methylthio and lower alkoxycarbonylamino Represents a lower alkyl which may have one substituent, a lower alkoxycarbonyl or a nitrogen-containing aliphatic heterocyclic group which may be substituted with oxo, or a lower alkoxy) or a hydrogen atom] or CONHR ⁹ ( wherein, R ⁹ is hydroxy, lower alkoxy, amino, lower alk Arylamino and have 1-2 substituents selected from the group consisting of di-lower alkylamino represents an even lower alkyl) represents,
R ⁵ represents aryl optionally having 1 to 3 substituents selected from the group consisting of halogen, hydroxy, lower alkoxy, nitro, amino, cyano and carboxy}
A therapeutic and / or prophylactic agent for glaucoma comprising the thiadiazoline derivative represented by the formula (I) or a pharmacologically acceptable salt thereof:

（式中、Ｒ¹、Ｒ²、Ｒ³、Ｒ⁴、Ｒ⁵およびｎはそれぞれ前記と同義である）で表されるチアジアゾリン誘導体である（１）記載の治療および／または予防剤。 (2) The following formula (II) in which the specific rotation at 20 ° C. with respect to the sodium D line (wavelength: 589.3 nm) when the thiadiazoline derivative is dissolved in methanol is negative

(Wherein R ¹ , R ² , R ³ , R ⁴ , R ⁵ and n are as defined above), the therapeutic and / or prophylactic agent according to (1).

(3) The therapeutic and / or prophylactic agent according to (1) or (2), wherein R ⁵ is phenyl.
(4) The therapeutic and / or prophylactic agent according to any one of (1) to (3), wherein R ³ is methyl, ethyl, isopropyl or tert-butyl.
(5) The therapeutic and / or prophylactic agent according to any one of (1) to (4), wherein R ¹ is a hydrogen atom.
(6) The therapeutic and / or prophylactic agent according to any one of (1) to (5), wherein R ² is methyl or tert-butyl.
(7) The therapeutic and / or prophylactic agent according to any one of (1) to (4), wherein R ¹ and R ² are combined to be trimethylene or tetramethylene.

(8) The therapeutic and / or prophylactic agent according to any one of (1) to (7), wherein R ⁴ is NHSO ₂ R ⁶ (wherein R ⁶ is as defined above).
(9) The therapeutic and / or prophylactic agent according to any one of (1) to (7), wherein R ⁴ is CONHR ⁹ (wherein R ⁹ is as defined above).
(10) The therapeutic and / or prophylactic agent according to any one of (1) to (9), wherein n is 1 or 2.

のいずれかで表されるチアジアゾリン誘導体である（２）記載の治療および／または予防剤。 (11) The thiadiazoline derivative has the following formulas (a) to (o)

The therapeutic and / or prophylactic agent according to (2), which is a thiadiazoline derivative represented by any one of:

で表される化合物またはその薬理学的に許容される塩を含有する緑内障の治療および／または予防剤。 (12) General formula (III)

A therapeutic and / or prophylactic agent for glaucoma comprising a compound represented by the formula: or a pharmacologically acceptable salt thereof.

  According to the present invention, a therapeutic and / or prophylactic agent for glaucoma containing a thiadiazoline derivative or a pharmacologically acceptable salt thereof as an active ingredient can be provided.

Hereinafter, the compound represented by general formula (I) and the compound represented by general formula (II) are referred to as compound (I) and compound (II), respectively. The same applies to the compounds of other formula numbers.
In the definition of each group of general formula (I) and general formula (II):
(I) Lower alkyl part of lower alkyl, lower alkoxy, lower alkylamino, di-lower alkylamino, lower alkoxycarbonyl and lower alkoxycarbonylamino includes, for example, linear or branched alkyl having 1 to 10 carbon atoms. More specifically, methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, pentyl, isopentyl, neopentyl, hexyl, heptyl, octyl, nonyl, decyl and the like can be mentioned. The two lower alkyl moieties of the di-lower alkylamino may be the same or different.

(Ii) Examples of lower alkenyl include linear or branched alkenyl having 2 to 10 carbon atoms, and more specifically vinyl, allyl, 1-propenyl, butenyl, pentenyl, hexenyl, heptenyl, octenyl, nonenyl. And decenyl.
(Iii) Aryl includes, for example, aryl having 6 to 14 carbon atoms, and more specifically includes phenyl, naphthyl and the like.
(Iv) Examples of alkylene include linear or branched alkylene having 1 to 10 carbon atoms, and more specifically, methylene, ethylene, trimethylene, tetramethylene, pentamethylene, hexamethylene, heptamethylene, octamethylene. , Nonamethylene, decamethylene, propylene, ethylethylene, methylmethylene, dimethylmethylene and the like.

(V) As the nitrogen-containing aliphatic heterocyclic group, for example, a 5-membered or 6-membered monocyclic aliphatic heterocyclic group containing at least one nitrogen atom, a bicyclic or tricyclic condensed 3- to 8-membered ring A cyclic aliphatic heterocyclic group containing at least one nitrogen atom, and more specifically, aziridinyl, azetidinyl, pyrrolidinyl, piperidino, piperidinyl, perhydroazepinyl, perhydroazosinyl, Examples include imidazolidinyl, pyrazolidinyl, piperazinyl, morpholino, morpholinyl, thiomorpholino, thiomorpholinyl, homopiperazinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, dihydroindolinyl, dihydroisoindolinyl and the like.

(Vi) Halogen means each atom of fluorine, chlorine, bromine and iodine.
In each group of compound (I) or (II),
R ¹ is preferably a hydrogen atom.
R ² is preferably methyl, ethyl, propyl, isopropyl, n-butyl, sec-butyl, tert-butyl and the like, more preferably methyl, tert-butyl and the like.
The alkylene formed by combining R ¹ and R ² is preferably trimethylene, tetramethylene, pentamethylene or the like.
R ³ is preferably methyl, ethyl, propyl, isopropyl, n-butyl, sec-butyl, tert-butyl and the like, more preferably methyl, ethyl, isopropyl, tert-butyl and the like.

R ⁴ is preferably NHSO ₂ R ^6B [wherein R ^6B is methyl, ethyl, propyl, vinyl, aminomethyl, 1-aminoethyl, 2-aminoethyl, 1-aminopropyl, 2-aminopropyl, 3 -Aminopropyl, methylaminomethyl, 1- (methylamino) ethyl, 2- (methylamino) ethyl, 1- (methylamino) propyl, 2- (methylamino) propyl, 3- (methylamino) propyl, dimethylamino Methyl, 1- (dimethylamino) ethyl, 2- (dimethylamino) ethyl, 1- (dimethylamino) propyl, 2- (dimethylamino) propyl, 3- (dimethylamino) propyl, ethylaminomethyl, 1- (ethyl Amino) ethyl, 2- (ethylamino) ethyl, 1- (ethylamino) propyl, 2- (ethylamino) Propyl, 3- (ethylamino) propyl, diethylaminomethyl, 1- (diethylamino) ethyl, 2- (diethylamino) ethyl, 1- (diethylamino) propyl, 2- (diethylamino) propyl, 3- (diethylamino) propyl, propylamino Methyl, 2- (propylamino) ethyl, 3- (propylamino) propyl, isopropylaminomethyl, 2- (isopropylamino) ethyl, 3- (isopropylamino) propyl and the like]

NHR ^7B [wherein R ^7B is a hydrogen atom, methyl, ethyl, propyl, isopropyl, n-butyl, aminomethyl, 1-aminoethyl, 2-aminoethyl, 1-aminopropyl, 2-aminopropyl, 3-amino. Propyl, methylaminomethyl, 1- (methylamino) ethyl, 2- (methylamino) ethyl, 1- (methylamino) propyl, 2- (methylamino) propyl, 3- (methylamino) propyl, dimethylaminomethyl, 1- (dimethylamino) ethyl, 2- (dimethylamino) ethyl, 1- (dimethylamino) propyl, 2- (dimethylamino) propyl, 3- (dimethylamino) propyl, ethylaminomethyl, 1- (ethylamino) Ethyl, 2- (ethylamino) ethyl, 3- (ethylamino) propyl, diethylaminomethyl, 1- (diethylamino) ethyl, 2- (diethylamino) ethyl, 3- (diethylamino) propyl, propylaminomethyl, 2- (propylamino) ethyl, 3- (propylamino) propyl, isopropylaminomethyl, 2- (isopropylamino) ) Represents ethyl, 3- (isopropylamino) propyl, etc.], CONHR ^9B [wherein R ^9B is methyl, ethyl, propyl, isopropyl, n-butyl, 2-hydroxyethyl, 2-hydroxypropyl, 3-hydroxypropyl. 2-hydroxy-n-butyl, 3-hydroxy-n-butyl, 4-hydroxy-n-butyl, 2-hydroxy-1- (hydroxymethyl) ethyl, 2-hydroxy-1-methylethyl, aminomethyl, 1 -Aminoethyl, 2-aminoethyl, 1-aminopropyl 2-aminopropyl, 3-aminopropyl, methylaminomethyl, 1- (methylamino) ethyl, 2- (methylamino) ethyl, 1- (methylamino) propyl, 2- (methylamino) propyl, 3- ( Methylamino) propyl, dimethylaminomethyl, 1- (dimethylamino) ethyl, 2- (dimethylamino) ethyl, 1- (dimethylamino) propyl, 2- (dimethylamino) propyl, 3- (dimethylamino) propyl, ethyl Aminomethyl, 1- (ethylamino) ethyl, 2- (ethylamino) ethyl, 3- (ethylamino) propyl, diethylaminomethyl, 1- (diethylamino) ethyl, 2- (diethylamino) ethyl, 3- (diethylamino) propyl , Propylaminomethyl, 2- (propylamino) ethyl, 3- (propyl Piruamino) propyl, isopropyl aminomethyl, 2- (isopropylamino) ethyl, 3- (isopropyl amino) propyl representing the like] and the like, more preferably in the NHSO ₂ R ^6B (wherein, R ^6B is as defined above And CONHR ^9B (wherein R ^9B has the same meaning as described above).

R ⁵ is preferably phenyl or the like.
n is preferably 1 or 2.

Furthermore, as compound (I) or (II), a compound in which the above-mentioned preferred substituents are combined is more preferred. For example, R ¹ is a hydrogen atom and R ² is methyl, ethyl, propyl, isopropyl, n-butyl, sec-butyl, tert-butyl, or the like, or R ¹ and R ^{2 taken} together are trimethylene, Tetramethylene, pentamethylene, etc., R ³ is methyl, ethyl, propyl, isopropyl, n-butyl, sec-butyl, tert-butyl, etc., R ⁴ is NHSO ₂ R ^6B (wherein R ^6B is NHR ^7B (wherein R ^7B is as defined above), CONHR ^9B (wherein R ^9B is as defined above), R ⁵ is phenyl, and n is A compound of 1 or 2 is preferred, R ¹ is a hydrogen atom, R ² is methyl, tert-butyl or the like, or R ¹ and R ² together represent trimethylene, tetramethylene, etc. ³ is methyl, ethyl, isopropyl, tert-butyl, etc., R ⁴ is NHSO ₂ R ^6B (wherein R ^6B is as defined above), CONHR ^9B (wherein R ^9B is as defined above) And the like, R ⁵ is phenyl, and n is 1 or 2, more preferably, R ¹ is a hydrogen atom, R ² is tert-butyl, or R ¹ and R ² are Together, it represents trimethylene, tetramethylene, etc., R ³ is methyl, ethyl, isopropyl, tert-butyl, etc., R ⁴ is NHSO ₂ R ^6B (wherein R ^6B is as defined above), etc. More preferred are compounds wherein R ⁵ is phenyl and n is 1 or 2.

In addition, the compound (I) is preferably a compound in which the specific optical rotation at 20 ° C. with respect to the sodium D line (wavelength: 589.3 nm) when dissolved in methanol shows a negative value.
Further, in compound (I), when n is 1, the asymmetric center to which R ⁵ is bonded is the R configuration, and when n is 2 or 3, the asymmetric center to which R ⁵ is bonded is the S configuration. Preferably there is.

  The pharmacologically acceptable salt of compound (I) includes, for example, a pharmacologically acceptable acid addition salt, metal salt, ammonium salt, organic amine addition salt, amino acid addition salt and the like. Examples of the pharmacologically acceptable acid addition salt of compound (I) include inorganic acid salts such as hydrochloride, sulfate, and phosphate, acetate, maleate, fumarate, citrate, and the like. Examples of the pharmacologically acceptable metal salt include organic acid salts and the like, such as alkali metal salts such as sodium salt and potassium salt, alkaline earth metal salts such as magnesium salt and calcium salt, aluminum salt and zinc salt. Examples of pharmacologically acceptable ammonium salts include ammonium and tetramethylammonium salts. Examples of pharmacologically acceptable organic amine addition salts include morpholine and piperidine. Examples of pharmacologically acceptable amino acid addition salts include lysine, glycine, phenylalanine, aspartic acid, and glutamate. Addition salts such as phosphate, and the like.

Next, the manufacturing method of compound (I) and (II) is demonstrated.
Manufacturing method 1
Compound (I) can be produced by the method described in WO2003 / 051854, WO2004 / 092147, WO2004 / 111024, and the like.
Manufacturing method 2
Compound (II) is obtained by, for example, separating an optical isomer separation column [eg, CHIRALPAK AD (manufactured by Daicel Chemical Industries, Ltd.)] It can manufacture by attaching | subjecting to preparative high performance liquid chromatography using, and resolving each stereoisomer.

（式中、Ｒ¹、Ｒ²、Ｒ³、Ｒ⁴、Ｒ⁵およびｎそれぞれ前記と同義である）

(Wherein R ¹ , R ² , R ³ , R ⁴ , R ⁵ and n are as defined above)

（式中、Ｒ¹、Ｒ²、Ｒ³、Ｒ⁴、Ｒ⁵およびｎそれぞれ前記と同義であり、Ｒ¹⁰は、１つの不斉中心を有する光学活性な置換基であり、例えば光学活性なＣ_1-10アルキル、光学活性なヒドロキシ置換Ｃ_1-10アルキル、光学活性なＣ_1-10アルコキシ置換Ｃ_1-10アルキル、光学活性なフェニル置換Ｃ_1-10アルキル、光学活性なナフチル置換Ｃ_1-10アルキルなどを表す。ここで、Ｃ_1-10アルキルおよびＣ_1-10アルコキシのＣ_1-10アルキル部分としては、例えば前記低級アルキルで例示した基があげられる） Production method 3
Compound (II) can also be produced according to the following steps.

Wherein R ¹ , R ² , R ³ , R ⁴ , R ⁵ and n are as defined above, and R ¹⁰ is an optically active substituent having one asymmetric center, for example, optically active C _1-10 alkyl, optically active hydroxy substituted C _1-10 alkyl, optically active C _1-10 alkoxy substituted C _1-10 alkyl, optically active phenyl substituted C _1-10 alkyl, optically active naphthyl substituted C _{1 -10} alkyl, etc. Here, examples of the C _1-10 alkyl moiety of C _1-10 alkyl and C _1-10 alkoxy include the groups exemplified for the lower alkyl)

An optically active acylating agent [R ¹⁰ COX (wherein R ¹⁰ is as defined above), a compound (A; racemate) obtained by the method described in WO2003 / 051854, WO2004 / 092147, WO2004 / 111024, etc. X represents a chlorine atom, a bromine atom, an iodine atom, etc.), (R ¹⁰ CO) ₂ O (wherein R ¹⁰ is as defined above), etc .; for example, (R)-(−)-2-phenylpropionyl Chloride, (S)-(+)-2-phenylpropionyl chloride, etc.], New Experimental Chemistry, Vol. 14, p. 1142 (1978), by reaction according to the method described in Maruzen Co., Ltd. to obtain a compound (B; diastereomer mixture) (step 1). Next, the obtained compound (B) is subjected to silica gel column chromatography, recrystallization and the like to divide each diastereomer to obtain a compound (C; one diastereomer) (step 2). Then, the obtained compound (C) is converted to the compound (D) according to the method described in WO2003 / 051854 (step 3), and finally acylation is performed according to the method described in WO2003 / 051854. (Step 4), compound (II) can be produced.

（式中、Ｒ^4aはＮＨＳＯ₂Ｒ⁶（式中、Ｒ⁶は前記と同義である）またはＮＨＲ⁷（式中、Ｒ⁷は前記と同義である）を表し、Ｒ¹、Ｒ²、Ｒ³およびＲ⁵はそれぞれ前記と同義である） Manufacturing method 4
Compound (II) wherein n is 1 and R ⁴ is NHSO ₂ R ⁶ (wherein R ⁶ is as defined above) or NHR ⁷ (wherein R ⁷ is as defined above) (IIa) can also be produced according to the following steps.

(Wherein R ^4a represents NHSO ₂ R ⁶ (wherein R ⁶ is as defined above) or NHR ⁷ (wherein R ⁷ is as defined above), R ¹ , R ² , R ³ and R ⁵ are as defined above)

  An optical isomer separation column [for example, CHIRALPAK AD (manufactured by Daicel Chemical Industries, Ltd.)] was used for the compound (Ib; racemate) obtained according to the method described in WO2003 / 051854, WO2004 / 092147, WO2004 / 111024, etc. Compound (Ic; one enantiomer) is obtained by preparative high performance liquid chromatography (Step 1). Subsequently, the obtained compound (Ic) is converted to the compound (Id) according to the method described in WO2004 / 111024 (step 2), and then the compound (Id) is converted according to the method described in WO2004 / 111024. By treatment (step 3), compound (IIa) can be produced.

Among compounds (I) and (II), there may exist stereoisomers such as positional isomers, geometric isomers, optical isomers, tautomers, etc., but the treatment of glaucoma of the present invention and For the prophylactic agent, all possible isomers and mixtures thereof can be used.
Compound (III) can be produced according to the method described in WO03 / 070701.
When it is desired to obtain a salt of compound (I), (II) or (III), it can be purified as it is when compound (I), (II) or (III) is obtained in the form of a salt. When obtained in the form, the compound (I), (II) or (III) may be dissolved or suspended in a suitable solvent, and a salt may be formed by adding an acid or base to be isolated and purified.

In addition, the compounds (I), (II) and (III) and pharmacologically acceptable salts thereof may exist in the form of adducts with water or various solvents. It can be used for the therapeutic and / or prophylactic agent for glaucoma of the present invention.
Specific examples of compounds (I) and (II) are shown in Tables 1 and 2. However, the compounds (I) and (II) used for the therapeutic and / or prophylactic agent for glaucoma of the present invention are not limited thereto.

Next, the pharmacological action of the compounds (I), (II) and (III) will be specifically described with reference to test examples.
Test Example 1: Cell Proliferation Inhibition Test for Human Vascular Endothelial Cells Normal human umbilical vein vascular endothelial cells (Kurashikibo, catalog number KE-4101) were used as human vascular endothelial cells. For cell culture, 2% fetal bovine serum (Kurashikibo, catalog number KE-6150), 10 ng / mL human recombinant epidermal growth factor (Kurashikibo, catalog number KE-6150), 5 ng / mL human group Reversible basic fibroblast growth factor (Kurashiki Spinning Co., Catalog No. KE-6150), 10 μg / mL heparin (Kurashiki Spinning Co., Catalog No. KE-6150), 1 μg / mL Hydrocortisone (Kurashiki Spinning Co., Ltd., Catalog No. KE) -6150), 50 μg / mL gentamicin (Kurashiki Spinning Co., catalog number KE-6150) and 50 ng / mL amphotericin B (Kurashiki Spinning Co., catalog number KE-6150), HuMedia-EB2 basal medium (Kurashiki Spinning Co., catalog number) KE-2150S) was used (hereinafter abbreviated as medium). The cells were cultured at 37 ° C. under 5% carbon dioxide conditions.

Normal human umbilical vein endothelial cells were seeded at 4000 cells / well (60 μL / well) in a 96-well plate (Nunk, catalog number 167008) and cultured overnight. 40 μL of a test compound diluted in a stepwise manner with a medium was added, followed by further culturing for 72 hours. Two 96-well plates were prepared, the first one immediately after addition of the test compound, the second one 72 hours after the addition, XTT of Cell Proliferation Kit II (XTT) (Roche Diagnostics, catalog number 1465015). 50 μL of the labeling mixture was added and incubated at 37 ° C. For each plate, the absorbance at 490 nm (control wavelength: 655 nm) was measured 4 hours after addition of XTT with a plate reader (Molecular Device, SpectraMax 340PC ³⁸⁴ ). Control cells treated with a solvent (dimethyl sulfoxide (DMSO)) instead of the test compound were grown in 72 hours (absorbance after 72 hours of solvent addition-absorbance at the time of solvent addition) as 100%. From the proliferation of the cells in the treated wells (absorbance after 72 hours of addition of test compound-absorbance at the time of addition of test compound), the proliferation rate at the time of addition of the test compound was calculated. A 50% growth inhibitory concentration GI ₅₀ value was calculated from a plot of the concentration of the test compound and the cell growth rate at that time.

Compounds 1, 2, a, b, d, e, h, j, 1, m, n, o and compound (III) inhibit GI ₅₀ value of 10 μmol / L or less against normal human umbilical vein endothelial cells Showed activity.
From this test, it is considered that compounds (I), (II) and compound (III) have cell growth inhibitory activity against human vascular endothelial cells such as normal human umbilical vein endothelial cells. It has been reported that the development of glaucoma involves the proliferation of vascular endothelial cells (Clin. Experimental Ophthalmology), Vol. 31, p. 492 (2003). (Year); Ophthalmologica, Vol. 218, p.344 (2004)), compounds (I), (II) and compound (III) are considered useful as therapeutic and / or prophylactic agents for glaucoma.

Test Example 2: Intraocular pressure reduction effect by instillation administration of compounds (I), (II) and (III) Japanese white rabbits are used. For intraocular pressure (IOP) measurements, eye drops are paralyzed with 2% lidocaine in the eyes of rabbits. A tonometer is used to monitor IOP. An ophthalmic solution containing 0.1% of compound (I), (II) or (III) is prepared and administered by 0.01-0.3 mL. Compound (I), (II) or (III) was measured by measuring IOP before and 0.5, 1, 6, 9 and 24 hours after administration of compound (I), (II) or (III). The intraocular pressure lowering effect by can be confirmed.

By this test, it can be confirmed that the compounds (I), (II) and (III) have an action of reducing intraocular pressure, and it can be confirmed that they are useful as a therapeutic and / or prophylactic agent for glaucoma.
Compound (I), (II) or (III) or a pharmacologically acceptable salt thereof can be administered alone as it is, but it is usually desirable to provide it as various pharmaceutical preparations. These pharmaceutical preparations are used for animals or humans.

The pharmaceutical preparation according to the present invention contains the compound (I), (II) or (III) or a pharmaceutically acceptable salt thereof alone or as a mixture with any other pharmaceutical ingredient as an active ingredient. be able to. In addition, these pharmaceutical preparations are produced by any method well known in the technical field of pharmaceutics by mixing the active ingredient together with one or more pharmaceutically acceptable carriers.
As the administration route, it is desirable to use one that is most effective in the treatment, and examples thereof include oral administration and parenteral administration such as intravenous administration.
Examples of the dosage form include tablets, injections, eye drops and the like.

Suitable for oral administration, such as tablets, excipients such as lactose and mannitol, disintegrants such as starch, lubricants such as magnesium stearate, binders such as hydroxypropylcellulose, surfactants such as fatty acid esters It can be manufactured using a plasticizer such as glycerin.
Formulations suitable for parenteral administration preferably comprise a sterile aqueous solution containing the active compound that is isotonic with the blood of the recipient. For example, in the case of an injection, a solution for injection is prepared using a carrier comprising a salt solution, a glucose solution, or a mixture of salt water and a glucose solution. For example, in the case of eye drops, the preparation is made using a buffering agent such as a phosphate buffer or boric acid buffer, an isotonic agent such as sodium chloride, a preservative such as benzalkonium chloride barabens, and the like.

Also in these parenteral agents, one kind selected from excipients, disintegrating agents, lubricants, binders, surfactants, plasticizers and diluents, preservatives, flavors and the like exemplified in the oral preparations. Or more auxiliary components can be added.
Compound (I), (II) or (III) or a pharmaceutically acceptable salt thereof is usually administered systemically or locally in an oral or parenteral form when used for the above purpose. . The dose and frequency of administration vary depending on the administration form, patient age, body weight, nature or severity of symptoms to be treated, etc., but usually oral, 0.01 to 1000 mg per adult, Preferably, the dose is in the range of 0.05 to 500 mg once to several times a day, or once every several days to 1 or 2 weeks. In the case of parenteral administration such as intravenous administration, it is usually 0.001 to 1000 mg, preferably 0.01 to 300 mg per adult, once to several times a day, or once every several days or every 1 to 3 weeks. Dosage once. Examples of the administration method include rapid intravenous injection, intravenous continuous administration in the range of 1 to 24 hours per day, and the like. In the case of eye drop administration, an eye drop containing usually 0.001 to 1% of compound (I), (II) or (III) or a pharmacologically acceptable salt thereof is once to several times a day or several times Instilled once daily or every 1 to 3 weeks. However, the dose and the number of doses vary depending on the various conditions described above.

Hereinafter, the present invention will be described in detail with reference to Examples and Reference Examples.
Proton nuclear magnetic resonance spectra ( ¹ H NMR) used in Reference Examples are those measured at 270 MHz or 300 MHz, and exchangeable protons may not be clearly observed depending on the compound and measurement conditions. In addition, although what is used normally is used as description of the multiplicity of a signal, br represents that it is an apparent wide signal.

Tablet (Compound 3)
A tablet having the following composition is prepared by a conventional method. Compound 3, 40 g, lactose 286.8 g and potato starch 60 g are mixed, and 10% aqueous solution of hydroxypropylcellulose 120 g is added thereto. The obtained mixture is kneaded by a conventional method, granulated and dried, and then sized to obtain granules for tableting. To this was added 1.2 g of magnesium stearate, mixed, and tableted with a tableting machine (RT-15 model, manufactured by Kikusui Co., Ltd.) having a 8 mm diameter punch to contain tablets (containing 20 mg of active ingredient per tablet). To get).
Formulation Compound 3 20 mg
Lactose 143.4 mg
Potato starch 30 mg
Hydroxypropylcellulose 6 mg
Magnesium stearate 0.6 mg
200 mg

Tablet (compound 4)
A tablet having the following composition is prepared by a conventional method. Compound 4, 40 g, lactose 286.8 g and potato starch 60 g are mixed, and 10% aqueous solution of hydroxypropylcellulose 120 g is added thereto. The obtained mixture is kneaded by a conventional method, granulated and dried, and then sized to obtain granules for tableting. To this was added 1.2 g of magnesium stearate, mixed, and tableted with a tableting machine (RT-15 model, manufactured by Kikusui Co., Ltd.) having a 8 mm diameter punch to contain tablets (containing 20 mg of active ingredient per tablet). To get).
Formulation Compound 4 20 mg
Lactose 143.4 mg
Potato starch 30 mg
Hydroxypropylcellulose 6 mg
Magnesium stearate 0.6 mg
200 mg

Tablet (Compound 7)
A tablet having the following composition is prepared by a conventional method. Compound 7, 40 g, lactose 286.8 g and potato starch 60 g are mixed, and 10% aqueous solution of hydroxypropylcellulose 120 g is added thereto. The obtained mixture is kneaded by a conventional method, granulated and dried, and then sized to obtain granules for tableting. To this was added 1.2 g of magnesium stearate, mixed, and tableted with a tableting machine (RT-15 model, manufactured by Kikusui Co., Ltd.) having a 8 mm diameter punch to contain tablets (containing 20 mg of active ingredient per tablet). To get).
Formulation Compound 7 20 mg
Lactose 143.4 mg
Potato starch 30 mg
Hydroxypropylcellulose 6 mg
Magnesium stearate 0.6 mg
200 mg

Injection (compound 3)
An injection having the following composition is prepared by a conventional method. Compound 3, 1 g and 5 g of D-mannitol are added to and mixed with distilled water for injection. Further, hydrochloric acid and aqueous sodium hydroxide solution are added to adjust the pH to 7, and then the total volume is made up to 1000 mL with distilled water for injection. . The obtained mixed solution is aseptically filled into glass vials by 2 mL to obtain an injection (containing 2 mg of active ingredient per vial).
Formulation Compound 3 2 mg
D-mannitol 10 mg
Hydrochloric acid appropriate amount Sodium hydroxide aqueous solution appropriate amount
Distilled water for injection
2.00 mL

Injection (Compound 9)
An injection having the following composition is prepared by a conventional method. Compound 9, 1 g and 5 g of D-mannitol are added to and mixed with distilled water for injection. Further, hydrochloric acid and sodium hydroxide aqueous solution are added to adjust the pH to 7, and then the total volume is made up to 1000 mL with distilled water for injection. . The obtained mixed solution is aseptically filled into glass vials by 2 mL to obtain an injection (containing 2 mg of active ingredient per vial).
Formulation Compound 9 2 mg
D-mannitol 10 mg
Hydrochloric acid appropriate amount Sodium hydroxide aqueous solution appropriate amount
Distilled water for injection
2.00 mL

Injection (Compound 12)
An injection having the following composition is prepared by a conventional method. Compound 12, 1 g, and 5 g of D-mannitol are added to and mixed with distilled water for injection. Further, hydrochloric acid and aqueous sodium hydroxide solution are added to adjust the pH to 7, and then the total volume is made up to 1000 mL with distilled water for injection. . The obtained mixed solution is aseptically filled into glass vials by 2 mL to obtain an injection (containing 2 mg of active ingredient per vial).
Formulation Compound 12 2 mg
D-mannitol 10 mg
Hydrochloric acid appropriate amount Sodium hydroxide aqueous solution appropriate amount
Distilled water for injection
2.00 mL

Tablet (compound a)
A tablet having the following composition is prepared by a conventional method. Compound a (40 g), lactose (286.8 g) and potato starch (60 g) are mixed, and 10% aqueous solution of hydroxypropylcellulose (120 g) is added thereto. The obtained mixture is kneaded by a conventional method, granulated and dried, and then sized to obtain granules for tableting. To this was added 1.2 g of magnesium stearate, mixed, and tableted with a tableting machine (RT-15 model, manufactured by Kikusui Co., Ltd.) having a 8 mm diameter punch to contain tablets (containing 20 mg of active ingredient per tablet). To get).
Formulation Compound a 20 mg
Lactose 143.4 mg
Potato starch 30 mg
Hydroxypropylcellulose 6 mg
Magnesium stearate 0.6 mg
200 mg

Tablet (Compound d)
A tablet having the following composition is prepared by a conventional method. Compound d (40 g), lactose (286.8 g) and potato starch (60 g) are mixed, and 10% aqueous solution of hydroxypropylcellulose (120 g) is added thereto. The obtained mixture is kneaded by a conventional method, granulated and dried, and then sized to obtain granules for tableting. To this was added 1.2 g of magnesium stearate, mixed, and tableted with a tableting machine (RT-15 model, manufactured by Kikusui Co., Ltd.) having a 8 mm diameter punch to contain tablets (containing 20 mg of active ingredient per tablet). To get).
Formulation Compound d 20 mg
Lactose 143.4 mg
Potato starch 30 mg
Hydroxypropylcellulose 6 mg
Magnesium stearate 0.6 mg
200 mg

Tablet (Compound e)
A tablet having the following composition is prepared by a conventional method. Compound e (40 g), lactose (286.8 g) and potato starch (60 g) are mixed, and 10% aqueous solution of hydroxypropylcellulose (120 g) is added thereto. The obtained mixture is kneaded by a conventional method, granulated and dried, and then sized to obtain granules for tableting. To this was added 1.2 g of magnesium stearate, mixed, and tableted with a tableting machine (RT-15 model, manufactured by Kikusui Co., Ltd.) having a 8 mm diameter punch to contain tablets (containing 20 mg of active ingredient per tablet). To get).
Formulation Compound e 20 mg
Lactose 143.4 mg
Potato starch 30 mg
Hydroxypropylcellulose 6 mg
Magnesium stearate 0.6 mg
200 mg

Tablet (Compound 1)
A tablet having the following composition is prepared by a conventional method. Compound l (40 g), lactose (286.8 g) and potato starch (60 g) are mixed, and 10% aqueous solution of hydroxypropylcellulose (120 g) is added thereto. The obtained mixture is kneaded by a conventional method, granulated and dried, and then sized to obtain granules for tableting. To this was added 1.2 g of magnesium stearate, mixed, and tableted with a tableting machine (RT-15 model, manufactured by Kikusui Co., Ltd.) having a 8 mm diameter punch to contain tablets (containing 20 mg of active ingredient per tablet). To get).
Formulation Compound l 20 mg
Lactose 143.4 mg
Potato starch 30 mg
Hydroxypropylcellulose 6 mg
Magnesium stearate 0.6 mg
200 mg

Tablet (compound m)
A tablet having the following composition is prepared by a conventional method. Compound m (40 g), lactose (286.8 g) and potato starch (60 g) are mixed, and 10% aqueous solution of hydroxypropylcellulose (120 g) is added thereto. The obtained mixture is kneaded by a conventional method, granulated and dried, and then sized to obtain granules for tableting. To this was added 1.2 g of magnesium stearate, mixed, and tableted with a tableting machine (RT-15 model, manufactured by Kikusui Co., Ltd.) having a 8 mm diameter punch to contain tablets (containing 20 mg of active ingredient per tablet). To get).
Formulation Compound m 20 mg
Lactose 143.4 mg
Potato starch 30 mg
Hydroxypropylcellulose 6 mg
Magnesium stearate 0.6 mg
200 mg

Injection (compound a)
An injection having the following composition is prepared by a conventional method. Compound a, 1 g and D-mannitol 5 g are added to and mixed with distilled water for injection. Further, hydrochloric acid and sodium hydroxide aqueous solution are added to adjust the pH to 7, and then the total volume is made up to 1000 mL with distilled water for injection. . The obtained mixed solution is aseptically filled into glass vials by 2 mL to obtain an injection (containing 2 mg of active ingredient per vial).
Formulation Compound a 2 mg
D-mannitol 10 mg
Hydrochloric acid appropriate amount Sodium hydroxide aqueous solution appropriate amount
Distilled water for injection
2.00 mL

Injection (compound l)
An injection having the following composition is prepared by a conventional method. Compound l, 1 g and 5 g of D-mannitol are added to and mixed with distilled water for injection. Further, hydrochloric acid and aqueous sodium hydroxide solution are added to adjust the pH to 7, and then the total volume is made up to 1000 mL with distilled water for injection. . The obtained mixed solution is aseptically filled into glass vials by 2 mL to obtain an injection (containing 2 mg of active ingredient per vial).
Formulation Compound l 2 mg
D-mannitol 10 mg
Hydrochloric acid appropriate amount Sodium hydroxide aqueous solution appropriate amount
Distilled water for injection
2.00 mL

Injection (compound m)
An injection having the following composition is prepared by a conventional method. Compound m, 1 g and D-mannitol 5 g are added to and mixed with distilled water for injection, and further adjusted to pH 7 by adding hydrochloric acid and aqueous sodium hydroxide solution, and the total volume is made up to 1000 mL with distilled water for injection. . The obtained mixed solution is aseptically filled into glass vials by 2 mL to obtain an injection (containing 2 mg of active ingredient per vial).
Formulation Compound m 2 mg
D-mannitol 10 mg
Hydrochloric acid appropriate amount Sodium hydroxide aqueous solution appropriate amount
Distilled water for injection
2.00 mL

Eye drops (compound 3)
An eye drop comprising the following composition is prepared by a conventional method. Compound 3, 1 g and sodium chloride 9 g are added to and mixed with distilled water for injection. Further, hydrochloric acid and aqueous sodium hydroxide solution are added to adjust the pH to 7, and then the total volume is made up to 1000 mL with distilled water for injection. The obtained mixed solution is aseptically filled into eyedrop containers 1 mL at a time to obtain eye drops (containing 1 mg of active ingredient per container).
Formulation Compound 3 1 mg
Sodium chloride 9 mg
Hydrochloric acid appropriate amount Sodium hydroxide aqueous solution appropriate amount
Distilled water for injection
1.00 mL

Eye drops (compound 4)
An eye drop comprising the following composition is prepared by a conventional method. Compound 4, 1 g and sodium chloride 9 g are added to and mixed with distilled water for injection. Further, hydrochloric acid and aqueous sodium hydroxide solution are added to adjust the pH to 7, and then the total volume is made up to 1000 mL with distilled water for injection. The obtained mixed solution is aseptically filled into eyedrop containers 1 mL at a time to obtain eye drops (containing 1 mg of active ingredient per container).
Formulation Compound 4 1 mg
Sodium chloride 9 mg
Hydrochloric acid appropriate amount Sodium hydroxide aqueous solution appropriate amount
Distilled water for injection
1.00 mL

Eye drops (compound a)
An eye drop comprising the following composition is prepared by a conventional method. Compound a, 1 g and sodium chloride 9 g are added to and mixed with distilled water for injection. Further, hydrochloric acid and aqueous sodium hydroxide are added to adjust the pH to 7, and then the total volume is made up to 1000 mL with distilled water for injection. The obtained mixed solution is aseptically filled into eyedrop containers 1 mL at a time to obtain eye drops (containing 1 mg of active ingredient per container).
Formulation Compound a 1 mg
Sodium chloride 9 mg
Hydrochloric acid appropriate amount Sodium hydroxide aqueous solution appropriate amount
Distilled water for injection
1.00 mL

Eye drops (compound d)
An eye drop comprising the following composition is prepared by a conventional method. Compound d (1 g) and sodium chloride (9 g) are added to and mixed with distilled water for injection. Further, hydrochloric acid and aqueous sodium hydroxide solution are added to adjust the pH to 7, and then the total volume is made up to 1000 mL with distilled water for injection. The obtained mixed solution is aseptically filled into eyedrop containers 1 mL at a time to obtain eye drops (containing 1 mg of active ingredient per container).
Formulation Compound d 1 mg
Sodium chloride 9 mg
Hydrochloric acid appropriate amount Sodium hydroxide aqueous solution appropriate amount
Distilled water for injection
1.00 mL

Eye drops (compound l)
An eye drop comprising the following composition is prepared by a conventional method. 1 g of compound 1 and 9 g of sodium chloride are added to and mixed with distilled water for injection. Further, hydrochloric acid and aqueous sodium hydroxide solution are added to adjust the pH to 7, and then the total volume is made up to 1000 mL with distilled water for injection. The obtained mixed solution is aseptically filled into eyedrop containers 1 mL at a time to obtain eye drops (containing 1 mg of active ingredient per container).
Formulation Compound l 1 mg
Sodium chloride 9 mg
Hydrochloric acid appropriate amount Sodium hydroxide aqueous solution appropriate amount
Distilled water for injection
1.00 mL

Eye drops (Compound m)
An eye drop comprising the following composition is prepared by a conventional method. Compound m (1 g) and sodium chloride (9 g) are added to and mixed with distilled water for injection. Further, hydrochloric acid and aqueous sodium hydroxide solution are added to adjust the pH to 7, and then the total volume is made up to 1000 mL with distilled water for injection. The obtained mixed solution is aseptically filled into eyedrop containers 1 mL at a time to obtain eye drops (containing 1 mg of active ingredient per container).
Formulation Compound m 1 mg
Sodium chloride 9 mg
Hydrochloric acid appropriate amount Sodium hydroxide aqueous solution appropriate amount
Distilled water for injection
1.00 mL

Reference Examples 1 to 13 (Compounds 1 to 13)
Compounds 1 to 13 were synthesized according to the methods described in WO2003 / 051854 or WO2004 / 111024, respectively.

Reference Example 14
Compound a: (-)-N- [4- (2,2-dimethylpropionyl) -5- (2-methanesulfonylaminoethyl) -5-phenyl-4,5-dihydro-1,3,4-thiadiazole- 2-yl] -2,2-dimethylpropanamide Step 1: (S)-(+)-2-phenylpropionic acid (4.88 g, 32.5 mmol) was dissolved in dichloromethane (20 mL) and thionyl chloride (30 mL) ) And stirred at room temperature for 4 hours. After the mixture was concentrated under reduced pressure, the obtained residue was dissolved in dichloromethane (10 mL) (dichloromethane solution). Next, N- {2- [5-amino-3- (2,2-dimethylpropionyl) -2-phenyl-2,3-dihydro-1,3,4-obtained according to the method described in WO2003 / 051854 Thiadiazol-2-yl] ethyl} methanesulfonamide (4.93 g, 12.8 mmol) was dissolved in dichloromethane (15 mL) and pyridine (3.1 mL), and the above dichloromethane solution was added. The mixture was stirred at room temperature for 1.5 hours, water was added, and the mixture was extracted with chloroform. The organic layer was washed with 1 mol / L hydrochloric acid, water and saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. Chloroform (50 mL) and diisopropyl ether (10 mL) were added to the residue and stirred. The precipitated powder was collected by filtration and then subjected to silica gel column chromatography (chloroform / acetone / n-hexane / ethyl acetate = 9/1/1). / 1, 9/1 / 6.5 / 3.5, 9/1/7/3, then 9/1/5/5), and N- [4- (2,2- One diastereomer of (dimethylpropionyl) -5- (2-methanesulfonylaminoethyl) -5-phenyl-4,5-dihydro-1,3,4-thiadiazol-2-yl] -2-phenylpropanamide ( 2.48 g, 38%), and N- [4- (2,2-dimethylpropionyl) -5- (2-methanesulfonylaminoethyl) -5-phenyl-4,5-dihydro- as the later eluting fraction 1,3,4-Chia Asia Was obtained Lumpur-2-yl] -2-phenyl-propanamide other diastereomer (2.80 g, 43%).

N- [4- (2,2-dimethylpropionyl) -5- (2-methanesulfonylaminoethyl) -5-phenyl-4,5-dihydro-1,3,4-thiadiazol-2-yl eluting first ] One diastereomer of 2-phenylpropanamide: ¹ H NMR (270 MHz, CDCl ₃ ) δ (ppm): 1.26 (s, 9H), 1.53 (d, J = 7.1 Hz, 3H), 2.60 ( m, 1H), 2.93 (s, 3H), 3.20 (m, 1H), 3.36 (m, 1H), 3.57 (m, 1H), 3.67 (q, J = 7.1 Hz, 1H), 4.45 (br t, 1H), 7.20-7.49 (m, 10H), 7.75 (s, 1H).
APCI-MS m / z: 515 (M−H) ^- .

N- [4- (2,2-dimethylpropionyl) -5- (2-methanesulfonylaminoethyl) -5-phenyl-4,5-dihydro-1,3,4-thiadiazol-2-yl eluting later ] The other diastereomer of 2-phenylpropanamide: ¹ H NMR (270 MHz, CDCl ₃ ) δ (ppm): 1.25 (s, 9H), 1.51 (d, J = 7.1 Hz, 3H), 2.56 (m, 1H), 2.96 (s, 3H), 3.23 (m, 1H), 3.37 (m, 1H), 3.62 (m, 1H), 3.63 (q, J = 7.1 Hz, 1H), 4.67 (br t , J = 5.9 Hz, 1H), 7.17-7.52 (m, 10H), 7.99 (s, 1H).
APCI-MS m / z: 515 (M−H) ^- .

Step 2: N- [4- (2,2-dimethylpropionyl) -5- (2-methanesulfonylaminoethyl) -5-phenyl-4,5-dihydro-1 eluting earlier obtained in Step 1 above , 3,4-thiadiazol-2-yl] -2-phenylpropanamide, one diastereomer (2.28 g, 4.41 mmol) was dissolved in methanol (100 mL), and cerium chloride heptahydrate (1.64 g) was dissolved. , 4.41 mmol) and sodium borohydride (6.68 g, 0.176 mmol) were added, and the mixture was stirred at room temperature for 40 minutes. To the mixture was added sodium borohydride (20.04 g, 0.5297 mmol) and methanol (250 mL) in three portions, and the mixture was further stirred at room temperature for 2 hours, and then concentrated under reduced pressure. Ethyl acetate and 1 mol / L hydrochloric acid were added to the residue, and the mixture was extracted with ethyl acetate. The organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (chloroform / acetone / n-hexane / ethyl acetate = 9/1/7/3 → 9/1/5/5). This operation was repeated, and the resulting crude product (total 0.802 g, 2.09 mmol) was dissolved in a mixed solvent of ethanol (20 mL) and n-hexane (200 mL), and the precipitated solid was filtered off. The filtrate is concentrated to give optically active N- {2- [5-amino-3- (2,2-dimethylpropionyl) -2-phenyl-2,3-dihydro-1,3,4-thiadiazole- 2-yl] ethyl} methanesulfonamide (0.647 g, 23%) was obtained.

Step 3: Optically active N- {2- [5-amino-3- (2,2-dimethylpropionyl) -2-phenyl-2,3-dihydro-1,3,4-obtained in Step 2 above Thiadiazol-2-yl] ethyl} methanesulfonamide (90 mg, 0.23 mmol) is dissolved in dichloromethane (4 mL), pyridine (0.224 mL, 2.77 mmol) and trimethylacetyl chloride (0.288 mL, 2.33 mmol) are added, Stir at room temperature for 3.5 hours. Water and 1 mol / L hydrochloric acid were added to the reaction mixture, and the mixture was extracted with ethyl acetate. The organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. After the residue was purified by silica gel column chromatography (n-hexane / ethyl acetate = 3/1 → 2/1), ethanol and then n-hexane were added to the resulting syrup, and the supernatant was separated by decantation and precipitated. A solid was obtained. Subsequently, the solid obtained by adding diisopropyl ether and stirring is pulverized, and compound a {(−)-N- [4- (2,2-dimethylpropionyl) -5- (2-methanesulfonylaminoethyl)- 5-phenyl-4,5-dihydro-1,3,4-thiadiazol-2-yl] -2,2-dimethylpropanamide} (60 mg, 55%) was obtained.

¹ H NMR (300 MHz, CDCl ₃ ) δ (ppm): 1.30 (s, 9H), 1.34 (s, 9H), 2.56-2.65 (m, 1H), 2.94 (s, 3H), 3.21-3.44 (m , 2H), 3.58-3.70 (m, 1H), 4.45 (br s, 1H), 7.28-7.37 (m, 5H), 7.97 (br s, 1H).
APCI-MS m / z: 467 (M-1) ^- .
Melting point: 204.0-206.0 ° C.
Specific rotation: The specific rotation at 20 ° C. with respect to sodium D line (wavelength: 589.3 nm) of a methanol solution of the obtained compound showed a value of −.

Reference Example 15
Compound b: (−)-N- [5- (2-methanesulfonylaminoethyl) -5-phenyl-4-propionyl-4,5-dihydro-1,3,4-thiadiazol-2-yl] -2, 2-Dimethylpropanamide Step 1: N- [2- (5-amino-2-phenyl-3-propionyl-2, obtained according to the method described in WO2003 / 051854 in the same manner as in Step 1 of Reference Example 14 3-dihydro-1,3,4-thiadiazol-2-yl) ethyl] methanesulfonamide (10.7 g, 30.0 mmol), and (R)-(−)-2-phenylpropionic acid (10.5 g, 69.9 mmol) And (R)-(−)-2-phenylpropionyl chloride prepared from thionyl chloride, N- [5- (2-methanesulfonylaminoethyl) -5-phenyl-4-propionyl-4,5 -Dihydro-1,3,4-thiadiazol-2-yl] -2-phenylpropanamide was obtained as a diastereomeric mixture (13.3 g, 92%). A part of this (3.89 g, 7.96 mmol) was purified by silica gel column chromatography (chloroform / acetonitrile / n-hexane / ethyl acetate = 9/1/1/1) to give N- One diastereomer of [5- (2-methanesulfonylaminoethyl) -5-phenyl-4-propionyl-4,5-dihydro-1,3,4-thiadiazol-2-yl] -2-phenylpropanamide (0.861 g, 22%), and N- [5- (2-methanesulfonylaminoethyl) -5-phenyl-4-propionyl-4,5-dihydro-1,3,4- as the first eluting fraction The other diastereomer (0.802 g, 20%) of thiadiazol-2-yl] -2-phenylpropanamide was obtained.

Step 2: In the same manner as in Step 2 of Reference Example 14, N- [5- (2-methanesulfonylaminoethyl) -5-phenyl-4-propionyl-4,5 eluted after obtained in Step 1 above. -Dihydro-1,3,4-thiadiazol-2-yl] -2-phenylpropanamide one diastereomer (4.41 g, 9.03 mmol), cerium chloride heptahydrate (3.37 g, 9.05 mmol) and From sodium borohydride (3.42 g, 90.5 mmol), N- [2- (5-amino-2-phenyl-3-propionyl-2,3-dihydro-1,3,4-thiadiazol-2-yl) ethyl Methanesulfonamide (2.16 g, 67%) was obtained.

Step 3: In the same manner as in Step 3 of Reference Example 14, N- [2- (5-amino-2-phenyl-3-propionyl-2,3-dihydro-1,3,4) obtained in Step 2 above was used. From thiadiazol-2-yl) ethyl] methanesulfonamide (0.0480 g, 0.135 mmol), pyridine (32.7 μL, 0.405 mmol) and trimethylacetyl chloride (41.7 μL, 0.338 mmol), compound b {(−)-N— [5- (2-Methanesulfonylaminoethyl) -5-phenyl-4-propionyl-4,5-dihydro-1,3,4-thiadiazol-2-yl] -2,2-dimethylpropanamide} (0.0504 g 84%).

¹ H NMR (270 MHz, CDCl ₃ ) δ (ppm): 1.13 (t, J = 6.0 Hz, 3H), 1.28 (s, 9H), 2.66 (m, 3H), 2.97 (s, 3H), 3.35 ( m, 2H), 3.61 (m, 1H), 4.58 (br s, 1H), 7.32 (m, 5H), 8.08 (br s, 1H).
APCI-MS m / z: 441 (M + 1) ⁺ .
Melting point: 107.0-110.0 ° C.
Specific rotation: The specific rotation at 20 ° C. with respect to sodium D line (wavelength: 589.3 nm) of a methanol solution of the obtained compound showed a value of −.

Reference Example 16
Compound c: (-)-N- {2- [3- (2,2-dimethylpropionyl) -5- (2-oxopyrrolidin-1-yl) -2-phenyl-2,3-dihydro-1,3 , 4-thiadiazol-2-yl] ethyl} methanesulfonamide N- {2- [5-amino-3- (2,2-dimethylpropionyl) -2-phenyl-2 obtained in Step 2 of Reference Example 14 3-Dihydro-1,3,4-thiadiazol-2-yl] ethyl} methanesulfonamide (0.647 g, 1.68 mmol) was dissolved in dichloromethane (25 mL) and pyridine (0.41 mL, 5.1 mmol) and 4- Bromobutyryl (0.49 mL, 4.2 mmol) was added and stirred at room temperature for 2 hours. Water was added to the reaction mixture, and the mixture was extracted with chloroform. The organic layer was washed with 0.5 mol / L hydrochloric acid and saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was dissolved in dimethyl sulfoxide (DMSO) (6 mL), sodium acetate (0.331 g, 4.04 mmol) was added, and the mixture was heated to 100 ° C. over 14 minutes with stirring. After allowing to cool, water was added to the mixture, and the mixture was extracted with ethyl acetate. The organic layer was washed with saturated brine and dried over anhydrous sodium sulfate. After concentration under reduced pressure, the residue was purified by flash column chromatography (n-hexane / ethyl acetate = 3/1 → 1/1) and then recrystallized from acetone to give compound c {(−)-N—. {2- [3- (2,2-dimethylpropionyl) -5- (2-oxopyrrolidin-1-yl) -2-phenyl-2,3-dihydro-1,3,4-thiadiazol-2-yl] Ethyl} methanesulfonamide} (0.649 g, 85%) was obtained.

¹ H NMR (270 MHz, CDCl ₃ ) δ (ppm): 1.34 (s, 9H), 2.23 (m, 2H), 2.56 (m, 2H), 2.61 (m, 1H), 2.97 (s, 3H), 3.27 (m, 1H), 3.40 (m, 1H), 3.63 (m, 1H), 3.98 (m, 2H), 4.01 (br t, J = 3.5 Hz, 1H), 7.20-7.37 (m, 5H).
APCI-MS m / z: 453 (M + 1) ⁺ .
Melting point: 107.0-110.0 ° C.
Specific rotation: The specific rotation at 20 ° C. with respect to sodium D line (wavelength: 589.3 nm) of a methanol solution of the obtained compound showed a value of −.

Reference Example 17
Compound d: (-)-N- [4-isobutyryl-5- (2-methanesulfonylaminoethyl) -5-phenyl-4,5-dihydro-1,3,4-thiadiazol-2-yl] -2, 2-Dimethylpropanamide Step 1: N- [4-isobutyryl-5- (2-methanesulfonylaminoethyl) -5-phenyl-4,5-dihydro-1,3 obtained according to the method described in WO2003 / 051854 , 4-thiadiazol-2-yl] -2,2-dimethylpropanamide (2.32 g, 5.10 mmol) by preparative high performance liquid chromatography (HPLC) [column: CHIRALPAK AD (manufactured by Daicel Chemical Industries); elution solvent: 12% isopropyl alcohol / n-hexane; flow rate: 6 mL / min; column temperature: 25 ° C.], and fractions having a retention time of 10.2 minutes and 11.2 minutes were fractionated. Among these, the fraction of 11.2 minutes was concentrated, and the residue was recrystallized from n-pentane and ethanol to give compound d {(−)-N- [4-isobutyryl-5- (2-methanesulfonylaminoethyl). -5-phenyl-4,5-dihydro-1,3,4-thiadiazol-2-yl] -2,2-dimethylpropanamide} (0.707 g, 30%) was obtained as white crystals.
¹ H NMR (270 MHz, CDCl ₃ ) δ (ppm): 1.15 (2 xd, J = 2.0 Hz, 6H), 1.29 (s, 9H), 2.57-2.67 (m, 1H), 2.96 (s, 3H) , 3.23-3.44 (m, 3H), 3.37-3.68 (m, 1H), 4.46 (br s, 1H), 7.25-7.38 (m, 5H), 8.00 (br s, 1H).
APCI-MS m / z: 453 (M-1) ^- .
Melting point: 162.0-164.0 ° C.
Specific rotation: The specific rotation at 20 ° C. with respect to sodium D line (wavelength: 589.3 nm) of a methanol solution of the obtained compound showed a value of −.

Reference Example 18
Compound e: (−)-N- {2- [5- (2-oxopyrrolidin-1-yl) -2-phenyl-3-propionyl-2,3-dihydro-1,3,4-thiadiazole-2- Yl] ethyl} methanesulfonamide N- [2- (5-amino-2-phenyl-3-propionyl-2,3-dihydro-1,3,4-thiadiazole-2] obtained in Step 2 of Reference Example 15 -Yl) ethyl] methanesulfonamide (1.01 g, 2.83 mmol) and pyridine (330 μL, 4.08 mmol) in dichloromethane (40 mL) and 4-bromobutyryl chloride (390 μL, 3.40 mmol) at 0 ° C. And stirred at room temperature for 2 hours. 1 mol / L hydrochloric acid was added to the mixture, and the mixture was extracted with chloroform. The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure. DMSO (10 mL) and sodium acetate (560 mg, 6.83 mmol) were added to the residue, and the mixture was stirred at 100 ° C. for 5 minutes. After cooling to room temperature, water and 1 mol / L hydrochloric acid were added to the mixture, and the mixture was extracted with ethyl acetate. The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (chloroform / methanol = 20/1) to give compound e {(−)-N- {2- [5- (2-oxopyrrolidin-1-yl) -2-phenyl]. -3-propionyl-2,3-dihydro-1,3,4-thiadiazol-2-yl] ethyl} methanesulfonamide} (878 mg, 73%) was obtained.

¹ H NMR (270 MHz, CDCl ₃ ) δ (ppm): 1.15 (t, J = 6.6 Hz, 3H), 2.22 (m, 2H), 2.55-2.67 (m, 3H), 2.94 (s, 3H), 3.31-3.47 (m, 4H), 3.61 (m, 1H), 3.91-3.98 (m, 2H), 5.0 (br s, 1H), 7.20-7.35 (m, 5H).
APCI-MS m / z: 423 (M−1) ^- .
Melting point: 188.0-191.0 ° C.
Specific rotation: The specific rotation at 20 ° C. with respect to sodium D line (wavelength: 589.3 nm) of a methanol solution of the obtained compound showed a value of −.

Reference Example 19
Compound f: (−)-N- [4-acetyl-5- (2-methanesulfonylaminoethyl) -5-phenyl-4,5-dihydro-1,3,4-thiadiazol-2-yl] -2, 2-Dimethylpropanamide Step 1: Methanesulfonamide (0.476 g, 5.00 mmol) is dissolved in N, N-dimethylformamide (DMF) (10 mL), and 60% sodium hydride (0.275 g, 5.00 mmol) is dissolved in 0. The mixture was added at 0 ° C. and stirred at the same temperature for 20 minutes. Subsequently, 3-chloropropiophenone (843 mg, 5.00 mol) was added, and the mixture was stirred at the same temperature for 2 hours, and further stirred at room temperature for 15 hours. Water was added to the mixture and extracted with ethyl acetate. The organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (chloroform / methanol = 20/1) to give N-methanesulfonyl-3-aminopropiophenone (240 mg, 21%).

Then, in the same manner as described in WO2003 / 051854, N-methanesulfonyl-3-aminopropiophenone (388 mg, 1.71 mmol) and thiosemicarbazide (156 mg, 1.71 mmol) obtained above were obtained from N-methane. Sulfonyl-3-aminopropiophenone = thiosemicarbazone (219 mg, 45%) was obtained.
Step 2: N-methanesulfonyl-3-aminopropiophenone = thiosemicarbazone (9.83 g, 32.7 mmol) obtained in Step 1 above is dissolved in acetic anhydride (38 mL) and stirred at 130 ° C. for 10 minutes. After that, the mixture was further stirred at 70 ° C. for 2 hours and then at room temperature for 5 hours. The precipitated solid was collected by filtration to give N- [4-acetyl-5- (2-methanesulfonylaminoethyl) -5-phenyl-4,5-dihydro-1,3,4-thiadiazol-2-yl] Acetamide (11.3 g, 73%) was obtained.

Step 3: In the same manner as described in WO2003 / 051854, N- [4-acetyl-5- (2-methanesulfonylaminoethyl) -5-phenyl-4,5-dihydro-obtained in Step 2 above. From 1,3,4-thiadiazol-2-yl] acetamide (5.22 g, 13.6 mmol), sodium borohydride (5.14 g, 136 mmol) and cerium chloride heptahydrate (5.07 g, 13.6 mmol), N -[2- (3-Acetyl-5-amino-2-phenyl-2,3-dihydro-1,3,4-thiadiazol-2-yl) ethyl] methanesulfonamide was obtained.

  It was then obtained with (R)-(−)-2-phenylpropionyl chloride prepared from (R)-(−)-2-phenylpropionic acid (4.65 g, 3.10 mmol) and thionyl chloride (30 mL). N- [2- (3-acetyl-5-amino-2-phenyl-2,3-dihydro-1,3,4-thiadiazol-2-yl) ethyl] methanesulfonamide and the process of Reference Example 14 In the same manner as in 1, it was treated in pyridine (5.0 mL, 61.8 mmol) and purified by silica gel column chromatography (chloroform / n-hexane / ethyl acetate / methanol = 20/3/2/1). N- [4-acetyl-5- (2-methanesulfonylaminoethyl) -5-phenyl-4,5-dihydro-1,3,4-thiadiazol-2-yl] -2-phenyl as a fraction eluted in Prop One diastereomer of amide (0.75 g, 12%) and N- [4-acetyl-5- (2-methanesulfonylaminoethyl) -5-phenyl-4,5-dihydro- as the later eluting fraction The other diastereomer (0.82 g, 13%) of 1,3,4-thiadiazol-2-yl] -2-phenylpropanamide was obtained.

Step 4: N- [4-acetyl-5- (2-methanesulfonylaminoethyl) -5-phenyl-4,5 eluted after obtained in Step 3 in the same manner as in Step 2 of Reference Example 14. -Dihydro-1,3,4-thiadiazol-2-yl] -2-phenylpropanamide, another diastereomer (0.632 g, 1.33 mmol), cerium chloride heptahydrate (0.496 g, 1.33 mmol) And sodium borohydride (0.503 g, 13.3 mmol), optically active N- [2- (3-acetyl-5-amino-2-phenyl-2,3-dihydro-1,3,4-thiadiazole-2 -Ill) ethyl] methanesulfonamide (232 mg, 51%) was obtained.

Step 5: In the same manner as in Step 3 of Reference Example 14, the optically active N- [2- (3-acetyl-5-amino-2-phenyl-2,3-dihydro-1, obtained in the above Step 4 was used. From 3,4-thiadiazol-2-yl) ethyl] methanesulfonamide (0.0393 g, 0.115 mmol), pyridine (44.7 μL, 0.552 mmol) and trimethylacetyl chloride (56.7 μL, 0.460 mmol), compound f {(−) -N- [4-acetyl-5- (2-methanesulfonylaminoethyl) -5-phenyl-4,5-dihydro-1,3,4-thiadiazol-2-yl] -2,2-dimethylpropanamide} (0.0420 g, 86%) was obtained.
¹ H NMR (270 MHz, CDCl ₃ ) δ (ppm): 1.28 (s, 9H), 2.30 (s, 3H), 2.55-2.68 (m, 1H), 2.97 (s, 3H), 3.30-3.43 (m , 2H), 3.59-3.68 (m, 1H), 4.44 (br s, 1H), 7.27-7.39 (m, 5H), 8.00 (br s, 1H).
APCI-MS m / z: 425 (M−1) ^- .
Melting point: 187.0-190.0 ° C.
Specific rotation: The specific rotation at 20 ° C. with respect to sodium D line (wavelength: 589.3 nm) of a methanol solution of the obtained compound showed a value of −.

Reference Example 20
Compound g: N- {2- [3-acetyl-5- (2-oxopyrrolidin-1-yl) -2-phenyl-2,3-dihydro-1,3,4-thiadiazol-2-yl] ethyl} Methanesulfonamide In the same manner as in Reference Example 16, N- [2- (3-acetyl-5-amino-2-phenyl-2,3-dihydro-1,3,4) obtained in Step 4 of Reference Example 19 was used. -Thiadiazol-2-yl) ethyl] methanesulfonamide (0.0300 g, 0.0876 mmol), pyridine (33.6 μL, 0.420 mmol), 4-bromobutyryl chloride (40.6 μL, 0.350 mmol) and sodium acetate (0.0575 g, 0.701 mmol) Compound g {N- {2- [3-acetyl-5- (2-oxopyrrolidin-1-yl) -2-phenyl-2,3-dihydro-1,3,4-thiadiazol-2-yl] Ethyl} methanesulfonamide } (0.0301 g, 84%).
¹ H NMR (270 MHz, CDCl ₃ ) δ (ppm): 2.15 (m, 2H), 2.33 (s, 3H), 2.50-2.67 (m, 3H), 2.97 (s, 3H), 3.31-3.44 (m , 2H), 3.60-3.65 (m, 1H), 3.87-3.97 (m, 2H), 4.46 (br s, 1H), 7.24-7.38 (m, 5H).
APCI-MS m / z: 409 (M-1) ^- .
Melting point: 137.0-140.0 ° C.

Reference Example 21
Compound h: (−)-N- {2- [3-acetyl-5- (2-oxopiperidino) -2-phenyl-2,3-dihydro-1,3,4-thiadiazol-2-yl] ethyl} methane Sulfonamide In the same manner as in Reference Example 16, N- [2- (3-acetyl-5-amino-2-phenyl-2,3-dihydro-1,3,4-] obtained in Step 4 of Reference Example 19 was used. From thiadiazol-2-yl) ethyl] methanesulfonamide (0.0260 g, 0.0759 mmol), pyridine (29.3 μL, 0.365 mmol), 5-bromovaleryl chloride (40.7 μL, 0.304 mmol) and sodium acetate (0.0498 g, 0.607 mmol) H ((−)-N- {2- [3-acetyl-5- (2-oxopiperidino) -2-phenyl-2,3-dihydro-1,3,4-thiadiazol-2-yl] ethyl} Methanesulfonamide} ( 0.0241 g, 75%).
¹ H NMR (270 MHz, CDCl ₃ ) δ (ppm): 1.82-1.98 (m, 4H), 2.33 (s, 3H), 2.52-2.62 (m, 3H), 2.95 (s, 3H), 3.27-3.38 (m, 2H), 3.59-3.70 (m, 1H), 3.84-3.92 (m, 2H), 4.62 (br s, 1H), 7.23-7.37 (m, 5H).
APCI-MS m / z: 423 (M−1) ^- .
Melting point: 169.0-171.0 ° C.
Specific rotation: The specific rotation at 20 ° C. with respect to sodium D line (wavelength: 589.3 nm) of a methanol solution of the obtained compound showed a value of −.

Reference Example 22
Compound i: N- {4- (2,2-dimethylpropionyl) -5- [2- (2-ethylaminoethanesulfonylamino) ethyl] -5-phenyl-4,5-dihydro-1,3,4 Thiadiazol-2-yl} -2,2-dimethylpropanamide Step 1: Palladium (II) acetate (125 mg, 0.559 mmol) and triphenylphosphine (317 mg, 1.21 mmol) in tetrahydrofuran (THF) (50 mL) Dissolved. N-tert-butoxycarbonyl-β-alanine (2.07 g, 10.9 mmol), phenylboronic acid (1.61 g, 13.2 mmol), distilled water (0.477 mL, 26.5 mmol) and trimethylacetic anhydride (3.23) were added to the resulting solution. mL, 15.9 mmol) was added, followed by stirring at 60 ° C. for 24 hours. The mixture was filtered, saturated aqueous sodium hydrogen carbonate solution was added to the filtrate, and the mixture was extracted with ethyl acetate. The organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (hexane / ethyl acetate = 9/1 → 4/1) to give (3-oxo-3-phenylpropyl) carbamic acid tert-butyl ester (1.85 g, 68%). Obtained.

Step 2: (3-Oxo-3-phenylpropyl) carbamic acid tert-butyl ester (513 mg, 2.06 mmol) obtained in Step 1 above was dissolved in methanol (40 mL). Thiosemicarbazide hydrochloride (562 mg, 4.40 mmol) was added to the resulting solution, and the mixture was stirred at room temperature for 8 hours. Water was added to the mixture and extracted with ethyl acetate. The organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to give a pale yellow solid (513 mg). Part of the obtained solid (198 mg) was dissolved in dichloromethane (10 mL). Pyridine (0.300 mL, 3.73 mmol) and trimethylacetyl chloride (0.415 mL, 3.37 mmol) were added to the resulting solution, and the mixture was stirred at room temperature for 22 hours. A saturated aqueous sodium hydrogen carbonate solution was added to the mixture, and the mixture was further stirred at room temperature for 1 hour, and then extracted with ethyl acetate. The organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by preparative silica gel thin layer chromatography (n-hexane / ethyl acetate = 2/1) to give {2- [3- (2,2-dimethylpropionyl) -5- (2,2-dimethyl Propionylamino) -2-phenyl-2,3-dihydro-1,3,4-thiadiazol-2-yl] ethyl} carbamic acid tert-butyl ester (319 mg, 100%) was obtained.
APCI-MS m / z: 491 (M + H) ⁺ .

Step 3: {2- [3- (2,2-dimethylpropionyl) -5- (2,2-dimethylpropionylamino) -2-phenyl-2,3-dihydro-1,3 obtained in Step 2 above , 4-Thiadiazol-2-yl] ethyl} carbamic acid tert-butyl ester (274 mg, 0.557 mmol) was dissolved in dichloromethane (10 mL). To the obtained solution was added trifluoroacetic acid (1.0 mL), and the mixture was stirred at room temperature for 3 hours. The mixture was concentrated under reduced pressure. Diisopropyl ether was added to the residue and stirred for 3 hours. The precipitated white solid was collected by filtration to give N- [5- (2-aminoethyl) -4- (2,2-dimethylpropionyl) -5-phenyl-4,5-dihydro-1,3,4- Thiadiazol-2-yl] -2,2-dimethylpropanamide trifluoroacetate (252 mg, 90%) was obtained.
APCI-MS m / z: 391 (M + H) ⁺ .

Step 4: N- [5- (2-Aminoethyl) -4- (2,2-dimethylpropionyl) -5-phenyl-4,5-dihydro-1,3,4-thiadiazole obtained in Step 3 above -2-yl] -2,2-dimethylpropanamide trifluoroacetate (0.25 g, 0.53 mmol) was dissolved in methanol (5 mL), and ion-exchanged silica gel [SCX (BONDESIL SCX 40 uM, manufactured by Varian)] On a column packed with After the SCX was washed with methanol, fractions eluted with 1% hydrogen chloride-methanol solution were collected and concentrated under reduced pressure to give N- [5- (2-aminoethyl) -4- (2,2-dimethyl). Propionyl) -5-phenyl-4,5-dihydro-1,3,4-thiadiazol-2-yl] -2,2-dimethylpropanamide hydrochloride (0.19 g) was obtained as a white solid.

The hydrochloride obtained above was dissolved in dichloromethane (10 mL), 2-chloroethylsulfamoyl chloride (0.14 mL, 2.2 mmol) and triethylamine (0.62 mL, 4.6 mmol) were added at 0 ° C., and at the same temperature. Stir for 4 hours and then at room temperature for 10 hours. Saturated aqueous sodium hydrogen carbonate solution was added to the mixture, and the mixture was extracted with ethyl acetate. The organic layer was washed with saturated aqueous ammonium chloride solution and saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by preparative silica gel thin layer chromatography (n-hexane / ethyl acetate = 2/1), and N- [4- (2,2-dimethylpropionyl) -5- (2-ethenesulfonylaminoethyl)- 5-Phenyl-4,5-dihydro-1,3,4-thiadiazol-2-yl] -2,2-dimethylpropanamide (0.17 g, 65%) was obtained.
¹ H NMR (300 MHz, CDCl ₃ ) δ (ppm): 1.30 (s, 9H), 1.32 (s, 9H), 2.48-2.62 (m, 1H), 3.10-3.64 (m, 3H), 4.45 (br t, J = 5.7 Hz, 1H), 5.95 (d, J = 9.6 Hz, 1H), 6.26 (d, J = 16.2 Hz, 1H), 6.52 (dd, J = 9.6, 16.2 Hz, 1H), 7.22- 7.37 (m, 5H), 7.91 (br s, 1H).

Step 5: N- [4- (2,2-dimethylpropionyl) -5- (2-ethenesulfonylaminoethyl) -5-phenyl-4,5-dihydro-1,3,4 obtained in Step 4 above -Thiadiazol-2-yl] -2,2-dimethylpropanamide (0.16 g, 0.33 mmol) was dissolved in acetonitrile (10 mL), 70% ethylamine aqueous solution (1.0 mL, 12 mmol) was added, and the mixture was stirred at room temperature for 3 hours. Stir. The reaction solution was concentrated under reduced pressure, and the residue was purified by preparative silica gel thin layer chromatography (chloroform / methanol / concentrated aqueous ammonia = 100/10/1), and N- {4- (2,2-dimethylpropionyl)- 5- [2- (2-Ethylaminoethanesulfonylamino) ethyl] -5-phenyl-4,5-dihydro-1,3,4-thiadiazol-2-yl} -2,2-dimethylpropanamide (0.15 g , 86%).

Step 6: N- {4- (2,2-dimethylpropionyl) -5- [2- (2-ethylaminoethanesulfonylamino) ethyl] -5-phenyl-4,5-dihydro obtained in Step 5 above -1,3,4-thiadiazol-2-yl} -2,2-dimethylpropanamide (0.15 g, 0.29 mmol) by preparative high performance liquid chromatography (HPLC) [column: CHIRALCEL OD φ20 X 250 mm (Daicel Chemical Manufactured by Kogyo Co., Ltd.); elution solvent: hexane / ethanol = 80/20 (containing 0.1% diethylamine); flow rate: 6.0 mL / min], fraction of 9.0 minutes out of 7.5 minutes and 9.0 minutes of retention time Was sorted. The collected fraction was concentrated to give compound i {N- {4- (2,2-dimethylpropionyl) -5- [2- (2-ethylaminoethanesulfonylamino) ethyl] -5-phenyl-4. , 5-dihydro-1,3,4-thiadiazol-2-yl} -2,2-dimethylpropanamide} (33 mg, 22%) was obtained as a white solid.
¹ H NMR (270 MHz, CDCl ₃ ) δ (ppm): 1.11 (t, J = 7.1 Hz, 3H), 1.30 (s, 9H), 1.33 (s, 9H), 2.67 (q, J = 7.1 Hz, 2H), 2.53-2.70 (m, 1H), 3.00-3.76 (m, 8H), 7.22-7.38 (m, 5H), 7.92 (br s, 1H).
APCI-MS m / z: 526 (M + H) ⁺ .

Reference Example 23
Compound j: N- [5-aminomethyl-4- (2,2-dimethylpropionyl) -5-phenyl-4,5-dihydro-1,3,4-thiadiazol-2-yl] -2,2-dimethyl Propanamide step 1: [3- (2,2-dimethylpropionyl) -5- (2,2-dimethylpropionylamino) -2-phenyl-2,3-dihydro-1 obtained according to the method described in WO2004 / 092147 , 3,4-thiadiazol-2-ylmethyl] carbamic acid tert-butyl ester (HPLC) [column: CHIRALPAK AD φ4.6 X 250 mm (manufactured by Daicel Chemical Industries); eluent: hexane / ethanol = 80/20; flow rate: 1.0 mL / min], and fractions of 5.76 minutes out of the fractions with retention times of 4.63 minutes and 5.76 minutes were collected to obtain [3- (2,2 -Dimethylpropionyl) -5- (2,2-dimethylpropionylamino) -2-phenyl-2,3-dihydro-1,3,4-thiadiazol-2-ylmethyl] carbamic acid tert-butyl ester was obtained.

Step 2: [3- (2,2-Dimethylpropionyl) -5- (2,2-dimethylpropionylamino) -2-phenyl-2,3-dihydro-1,3,4-obtained in Step 1 above Thiadiazol-2-ylmethyl] carbamic acid tert-butyl ester (5.91 g, 12.4 mmol) was dissolved in ethyl acetate (20 mL), and then a 1 mol / L hydrogen chloride-ethyl acetate solution (40 mL) was added. Stir for 1 hour. The precipitated crystals were collected by filtration, and the obtained crystals were heated and dried under reduced pressure to give compound j {N- [5-aminomethyl-4- (2,2-dimethylpropionyl) -5-phenyl-4,5. -Dihydro-1,3,4-thiadiazol-2-yl] -2,2-dimethylpropanamide} hydrochloride (4.72 g, 92%) was obtained.
APCI-MS m / z: 377 (M + H) ⁺ .
Melting point: 175.0-182.0 ° C.

Reference Example 24
Compound k: N- [4- (2,2-dimethylpropionyl) -5-ethenesulfonylaminomethyl-5-phenyl-4,5-dihydro-1,3,4-thiadiazol-2-yl] -2,2 -Dimethylpropanamide Compound j {N- [5-aminomethyl-4- (2,2-dimethylpropionyl) -5-phenyl-4,5-dihydro-1,3,4-thiadiazole obtained in Reference Example 23 -2-yl] -2,2-dimethylpropanamide} hydrochloride (0.502 g, 1.22 mmol) was dissolved in ethyl acetate (20 mL), and chloroethanesulfonyl chloride (0.203 mL, 1.22 mmol) was added. Stir for minutes. The mixture was cooled to 0 ° C., triethylamine (0.680 mL, 4.88 mmol) was added, and the mixture was stirred at the same temperature for 30 min. Water and 1.0 mol / L hydrochloric acid were added to the mixture, and the mixture was extracted with chloroform. The organic layer was washed with water and saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by preparative silica gel thin layer chromatography (hexane / ethyl acetate = 3/2) to give compound k {N- [4- (2,2-dimethylpropionyl) -5-ethenesulfonylaminomethyl-5. -Phenyl-4,5-dihydro-1,3,4-thiadiazol-2-yl] -2,2-dimethylpropanamide} (0.408 g, 72%) was obtained.
¹ H NMR (300 MHz, CDCl ₃ ) δ (ppm): 1.29 (s, 9H), 1.33 (s, 9H), 3.85 (dd, J = 13.5, 4.8 Hz, 1H), 4.49 (dd, J = 13.5 , 8.1 Hz, 1H), 5.29 (br s, 1H), 5.93 (br d, J = 9.9 Hz, 1H), 6.27 (br d, J = 16.5 Hz, 1H), 6.53 (br dd, J = 16.4, 9.6 Hz, 1H), 7.27-7.34 (m, 5H), 8.06 (br s, 1H).
APCI-MS m / z: 466 (M) ⁺ .

Reference Example 25
Compound l: (-)-N- [4- (2,2-dimethylpropionyl) -5- (2-ethylaminoethanesulfonylaminomethyl) -5-phenyl-4,5-dihydro-1,3,4- Thiadiazol-2-yl] -2,2-dimethylpropanamide Compound k {N- [4- (2,2-dimethylpropionyl) -5-ethenesulfonylaminomethyl-5-phenyl-] obtained in Reference Example 24 4,5-dihydro-1,3,4-thiadiazol-2-yl] -2,2-dimethylpropanamide} (1.50 g, 3.21 mmol) was dissolved in acetonitrile (60 mL) and then 70% aqueous ethylamine solution. (13.9 mL) was added, and the mixture was stirred at room temperature for 1 hour. The mixture was concentrated under reduced pressure, and the resulting residue was dissolved in ethanol. Water was added and the precipitated solid was collected by filtration to give compound l {(−)-N- [4- (2,2-dimethylpropionyl) -5- (2-ethylaminoethanesulfonylaminomethyl) -5- Phenyl-4,5-dihydro-1,3,4-thiadiazol-2-yl] -2,2-dimethylpropanamide} (0.830 g, 51%) was obtained.

¹ H NMR (300 MHz, CDCl ₃ ) δ (ppm): 1.09 (t, J = 7.2 Hz, 3H), 1.28 (s, 9H), 1.34 (s, 9H), 2.64 (q, J = 7.2 Hz, 2H), 3.08 (m, 2H), 3.20 (m, 2H), 4.00 (d, J = 13.5 Hz, 1H), 4.57 (d, J = 13.5 Hz, 1H), 7.26-7.38 (m, 5H), 7.92 (br s, 1H).
APCI-MS m / z: 512 (M + 1) ⁺ .
Melting point: 169.0-171.0 ° C.
Specific rotation: The specific rotation at 20 ° C. with respect to sodium D line (wavelength: 589.3 nm) of a methanol solution of the obtained compound showed a value of −.

Reference Example 26
Compound m: (−)-N- [5- (2-dimethylaminoethanesulfonylaminomethyl) -4- (2,2-dimethylpropionyl) -5-phenyl-4,5-dihydro-1,3,4 Thiadiazol-2-yl] -2,2-dimethylpropanamide Step 1: N- [4- (2,2-dimethylpropionyl)-obtained according to the method described in WO2003 / 051854 in the same manner as in Reference Example 25. 5-ethenesulfonylaminomethyl-5-phenyl-4,5-dihydro-1,3,4-thiadiazol-2-yl] -2,2-dimethylpropanamide (0.05 g, 0.11 mmol) and 2 mol / L dimethyl From an amine-methanol solution (0.10 mL), N- [5- (2-dimethylaminoethanesulfonylaminomethyl) -4- (2,2-dimethylpropionyl) -5-pheny -4,5-dihydro-1,3,4-thiadiazol-2-yl] -2,2-dimethylpropanamide (0.02 g, 35%) was obtained.

Step 2: N- [5- (2-dimethylaminoethanesulfonylaminomethyl) -4- (2,2-dimethylpropionyl) -5-phenyl-4,5-dihydro-1,3 obtained in Step 1 above , 4-thiadiazol-2-yl] -2,2-dimethylpropanamide (50 mg) preparative high performance liquid chromatography (HPLC) [column: CHIRALPAK AD φ20 X 250 mm (manufactured by Daicel Chemical Industries); elution solvent : Hexane / ethanol = 91/9; flow rate: 5.0 mL / min], and fractions having a retention time of 22 minutes and 33 minutes were collected. Among these, by concentrating the fraction of 33 minutes, compound m {(−)-N- [5- (2-dimethylaminoethanesulfonylaminomethyl) -4- (2,2-dimethylpropionyl) -5- Phenyl-4,5-dihydro-1,3,4-thiadiazol-2-yl] -2,2-dimethylpropanamide} (17 mg, 34%) was obtained.

¹ H NMR (300 MHz, CDCl ₃ ) δ (ppm): 1.28 (s, 9H), 1.34 (s, 9H), 2.25 (s, 6H), 2.73 (br q, J = 6.3 Hz, 1H), 2.84 (br q, J = 6.2 Hz, 1H), 3.18 (br t, J = 6.6 Hz, 2H), 4.02 (d, J = 13.2 Hz, 1H), 4.58 (d, J = 13.2 Hz, 1H), 5.85 (br s, 1H), 7.27-7.35 (m, 5H), 8.02 (br s, 1H).
APCI-MS m / z: 512 (M + 1) ⁺ .
Melting point: 101.0-104.0 ° C.
Specific rotation: The specific rotation at 20 ° C. with respect to sodium D line (wavelength: 589.3 nm) of a methanol solution of the obtained compound showed a value of −.

Reference Example 27
Compound n: (-)-N- [5- (3-dimethylaminopropanesulfonylaminomethyl) -4- (2,2-dimethylpropionyl) -5-phenyl-4,5-dihydro-1,3,4- Thiadiazol-2-yl] -2,2-dimethylpropanamide Step 1: Compound j {N- [5-aminomethyl-4- (2,2-dimethylpropionyl) -5-phenyl] obtained in Reference Example 23 -4,5-dihydro-1,3,4-thiadiazol-2-yl] -2,2-dimethylpropanamide} hydrochloride (1.00 g, 2.42 mmol) was suspended in dichloromethane (25 mL) and cooled with ice. , Triethylamine (1.35 mL, 9.69 mmol) and 3-chloropropanesulfonyl chloride (0.442 mL, 3.63 mmol) were added, and the mixture was stirred at room temperature for 22 hours. Water and 1 mol / L hydrochloric acid were added to the mixture, and the mixture was extracted with chloroform. The organic layer was washed with saturated aqueous sodium hydrogen carbonate solution and saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. By triturating the residue with a mixed solvent of diisopropyl ether and ethyl acetate, N- [5- (3-chloropropanesulfonylaminomethyl) -4- (2,2-dimethylpropionyl) -5-phenyl-4,5 -Dihydro-1,3,4-thiadiazol-2-yl] -2,2-dimethylpropanamide (0.880 g, 70%) was obtained.
¹ H NMR (270 MHz, CDCl ₃ ) δ (ppm): 1.29 (s, 9H), 1.35 (s, 9H), 2.25 (m, 2H), 3.22 (m, 2H), 3.63 (m, 2H), 4.01 (dd, J = 5.1, 13.7 Hz, 1H), 4.60 (dd, J = 8.0, 13.7 Hz, 1H), 5.19 (dd, J = 5.1, 8.0 Hz, 1H), 7.23-7.41 (m, 5H) , 7.94 (s, 1H).
ESI-MS m / z: 515, 517 (M−H) ^- .

Step 2: N- [5- (3-Chloropropanesulfonylaminomethyl) -4- (2,2-dimethylpropionyl) -5-phenyl-4,5-dihydro-1,3,4 obtained in Step 1 above -Thiadiazol-2-yl] -2,2-dimethylpropanamide (1.50 g, 2.90 mmol), sodium iodide (8.69 g, 58.0 mmol) and sodium azide (1.89 g, 29.0 mmol) in DMF (20 mL) and stirred at 90 ° C. for 4 hours. Water was added to the mixture and extracted with ethyl acetate. The organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was triturated with diethyl ether to give N- [5- (3-azidopropanesulfonylaminomethyl) -4- (2,2-dimethylpropionyl) -5-phenyl-4,5-dihydro-1, 3,4-thiadiazol-2-yl] -2,2-dimethylpropanamide (1.82 g) was obtained.

The resulting N- [5- (3-azidopropanesulfonylaminomethyl) -4- (2,2-dimethylpropionyl) -5-phenyl-4,5-dihydro-1,3,4-thiadiazol is then obtained. -2-yl] -2,2-dimethylpropanamide was dissolved in THF (53 mL), water (10.6 mL) and triphenylphosphine (1.24 g, 4.73 mmol) were added, and the mixture was stirred at room temperature for 16 hours. The mixture was concentrated under reduced pressure, water and saturated aqueous sodium hydrogen carbonate solution were added, and the mixture was extracted with ethyl acetate. The organic layer was extracted with an aqueous hydrochloric acid solution, basified with a saturated aqueous sodium hydrogen carbonate solution, and then extracted with ethyl acetate. The obtained organic layer was concentrated under reduced pressure to give N- [5- (3-aminopropanesulfonylaminomethyl) -4- (2,2-dimethylpropionyl) -5-phenyl-4,5-dihydro-1 , 3,4-thiadiazol-2-yl] -2,2-dimethylpropanamide (1.29 g, 89%).
¹ H NMR (300 MHz, CDCl ₃ ) δ (ppm): 1.29 (s, 9H), 1.33 (s, 9H), 1.96 (m, 2H), 2.85 (t, J = 6.6 Hz, 2H), 3.19 ( t, J = 7.5 Hz, 2H), 3.99 (d, J = 13.7 Hz, 1H), 4.61 (d, J = 13.7 Hz, 1H), 7.24-7.39 (m, 5H).
APCI-MS m / z: 498 (M + H) ⁺ .

Step 3: N- [5- (3-aminopropanesulfonylaminomethyl) -4- (2,2-dimethylpropionyl) -5-phenyl-4,5-dihydro-1,3, obtained in Step 2 above 4-thiadiazol-2-yl] -2,2-dimethylpropanamide (1.00 g, 2.01 mmol) was dissolved in dichloroethane (40 mL), 37% formalin aqueous solution (1.63 mL, 0.201 mmol), acetic acid (1.15 mL, 20.1 mmol) and sodium triacetoxyborohydride (4.26 g, 20.1 mmol) were added, and the mixture was stirred at room temperature for 13 hours. Water and saturated aqueous sodium hydrogen carbonate solution were added to the mixture, and the mixture was extracted with chloroform. The organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (chloroform / methanol = 9/1 → 4/1 → 7/3) to obtain compound n {(−)-N- [5- (3-dimethylaminopropanesulfonylaminomethyl). ) -4- (2,2-dimethylpropionyl) -5-phenyl-4,5-dihydro-1,3,4-thiadiazol-2-yl] -2,2-dimethylpropanamide} (0.910 mg, 86% )

¹ H NMR (270 MHz, CDCl ₃ ) δ (ppm): 1.29 (s, 9H), 1.33 (s, 9H), 1.96 (m, 2H), 2.20 (s, 6H), 2.36 (t, J = 6.7 Hz, 2H), 3.12 (m, 2H), 3.96 (d, J = 13.4 Hz, 1H), 4.59 (m, 1H), 5.57 (br, 1H), 7.23-7.38 (m, 5H), 7.96 (br , 1H).
APCI-MS m / z: 526 (M + H) ⁺ .
Melting point: 92.0-95.0 ° C.
Specific rotation: The specific rotation at 20 ° C. with respect to sodium D line (wavelength: 589.3 nm) of a methanol solution of the obtained compound showed a value of −.

Reference Example 28
Compound o: 4- [3- (2,2-dimethylpropionyl) -5- (2,2-dimethylpropionylamino) -2-phenyl-2,3-dihydro-1,3,4-thiadiazol-2-yl ] -N- (2-hydroxyethyl) butanamide Step 1: 4- [3- (2,2-dimethylpropionyl) obtained according to the method described in WO2003 / 051854 in the same manner as described in WO2003 / 051854 -5- (2,2-dimethylpropionylamino) -2-phenyl-2,3-dihydro-1,3,4-thiadiazol-2-yl] butanoic acid methyl ester (11.2 g, 25.9 mmol) and borohydride From sodium (2.94 g, 77.6 mmol), 4- [5-amino-3- (2,2-dimethylpropionyl) -2-phenyl-2,3-dihydro-1,3 - to obtain a thiadiazol-2-yl] butanoic acid methyl ester (1.54 g, 17%).
APCI-MS m / z: 364 (M + H) ⁺ .

Step 2: In the same manner as in Step 1 of Reference Example 14, 4- [5-amino-3- (2,2-dimethylpropionyl) -2-phenyl-2,3-dihydro-1 obtained in Step 1 above. , 3,4-thiadiazol-2-yl] butanoic acid methyl ester (1.54 g, 4.24 mmol), (S)-(+)-2-phenylpropionic acid (1.99 g, 13.2 mmol), thionyl chloride (20 mL) A diastereomeric mixture was obtained from pyridine (1.80 mL, 22.0 mmol). The obtained diastereo mixture was purified by silica gel column chromatography (chloroform / acetone = 60/12), so that N- [3- (2,2-dimethylpropionyl) -2-phenyl was used as the fraction to be eluted first. One diastereomer (0.679 g, 32%) of methyl ester of -5- (2-phenylpropionylamino) -2,3-dihydro-1,3,4-thiadiazol-2-yl] butanoic acid was obtained.
¹ H NMR (300 MHz, CDCl ₃ ) δ (ppm): 1.24 (s, 9H), 1.54 (d, J = 8.0 Hz, 3H), 1.42-1.67 (m, 1H), 1.99-2.15 (m, 1H ), 2.20-2.32 (m, 1H), 2.38-2.46 (m, 2H), 3.03-3.16 (m, 1H), 3.62-3.71 (m, 1H), 3.67 (s, 3H), 7.18-7.47 (m , 10H), 7.64 (br s, 1H).
APCI-MS m / z: 496 (M + H) ⁺ .

Step 3: Sodium hydroxide (0.240 g, 6.01 mmol) was dissolved in water (4.0 mL), and then dioxane (8.0 mL) was added and stirred. N- [3- (2,2-dimethylpropionyl) -2-phenyl-5- (2-phenylpropionylamino) -2,3-dihydro-1,3,4 obtained in the above step 2 was added to the obtained solution. -Thiadiazol-2-yl] butanoic acid One diastereomer (0.992 g, 2.00 mmol) of methyl ester was added and stirred at room temperature for 5 hours. To the mixture were added 1 mol / L hydrochloric acid (20 mL) and water (30 mL), and the precipitated white solid was collected by filtration. The obtained solid was washed with water and diisopropyl ether and dried under reduced pressure to give 4- [3- (2,2-dimethylpropionyl) -2-phenyl-5- (2-phenylpropionylamino) -2,3. -Dihydro-1,3,4-thiadiazol-2-yl] butanoic acid (9.60 g, 99%) was obtained.
APCI-MS m / z: 481 (M + H) ⁺ .

Step 4: 4- [3- (2,2-Dimethylpropionyl) -2-phenyl-5- (2-phenylpropionylamino) -2,3-dihydro-1,3,4-thiadiazole-obtained above To 2-yl] butanoic acid (1.03 g, 2.14 mmol), oxalyl chloride (0.223 mL, 2.57 mmol) and DMF (17 μL, 0.214 mmol) were added at 0 ° C., and the mixture was stirred at the same temperature for 1 hour. The mixture was concentrated under reduced pressure, dichloromethane (20 mL) was added to the residue, and the mixture was stirred at 0 ° C. Ethanolamine (1.2 mL, 21.4 mmol) was added, and the mixture was stirred at room temperature for 3 hr. 1 mol / L hydrochloric acid (20 mL) and water (30 mL) were added to the mixture, and the mixture was extracted with chloroform. The organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. Diisopropyl ether was added to the resulting residue, and the precipitated white solid was collected by filtration. The obtained solid was washed with water and diisopropyl ether and then dried under reduced pressure to give 4- [3- (2,2-dimethylpropionyl) -2-phenyl-5- (2-phenylpropionylamino) -2, 3-Dihydro-1,3,4-thiadiazol-2-yl] -N- (2-hydroxyethyl) butanamide (1.10 g, 99%) was obtained.
APCI-MS m / z: 525 (M + H) ⁺ .

Step 5: 4- [3- (2,2-Dimethylpropionyl) -2-phenyl-5- (2-phenylpropionylamino) -2,3-dihydro-1,3,4-thiadiazole obtained in Step 4 above -2-yl] -N- (2-hydroxyethyl) butanamide (1.21 g, 2.31 mmol) was added with dichloromethane (20 mL), stirred at 0 ° C., pyridine (0.470 mL, 5.77 mmol) and tert-chloride. Butyldimethylsilyl (869 mg, 5.77 mmol) was added, and the mixture was stirred at room temperature for 3 hours. 1 mol / L hydrochloric acid (20 mL) and water (30 mL) were added to the mixture, and the mixture was extracted with chloroform. The organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. Diisopropyl ether was added to the resulting residue, and the precipitated white solid was collected by filtration. The obtained solid was washed with water and diisopropyl ether and then dried under reduced pressure to give N- [2- (tert-butyldimethylsiloxy) ethyl] -4- [3- (2,2-dimethylpropionyl) -2. -Phenyl-5- (2-phenylpropionylamino) -2,3-dihydro-1,3,4-thiadiazol-2-yl] butanamide (1.25 g, 85%) was obtained.
APCI-MS m / z: 638 (M + H) ⁺ .

Step 6: In the same manner as in Step 2 of Reference Example 14, N- [2- (tert-butyldimethylsiloxy) ethyl] -4- [3- (2,2-dimethylpropionyl)-obtained in Step 5 above 2-Phenyl-5- (2-phenylpropionylamino) -2,3-dihydro-1,3,4-thiadiazol-2-yl] butanamide (0.376 g, 0.588 mmol) and sodium borohydride (0.111 g, 2.94 mmol) from 4- [5-amino-3- (2,2-dimethylpropionyl) -2-phenyl-2,3-dihydro-1,3,4-thiadiazol-2-yl] -N- [2- (Tert-Butyldimethylsiloxy) ethyl] butanamide (0.113 g, 38%) was obtained.
¹ H NMR (270 MHz, CDCl ₃ ) δ (ppm): 0.03 (s, 3H), 0.07 (s, 3H), 0.86 (s, 9H), 0.90 (s, 9H), 2.15-2.28 (m, 1H ), 2.49-2.58 (m, 1H), 2.62-2.82 (m, 2H), 3.07-3.13 (m, 1H), 3.27-3.47 (m, 3H), 3.59-3.72 (m, 2H), 4.21 (br s, 2H), 5.97 (m, 1H), 7.22-7.44 (m, 5H).
APCI-MS m / z: 507 (M + H) ⁺ .

Step 7: In the same manner as in Step 3 of Reference Example 14, 4- [5-amino-3- (2,2-dimethylpropionyl) -2-phenyl-2,3-dihydro-1 obtained in Step 6 above. , 3,4-thiadiazol-2-yl] -N- [2- (tert-butyldimethylsiloxy) ethyl] butanamide (0.0683 g, 0.135 mmol), pyridine (131 μL, 1.62 mmol) and trimethylacetyl chloride (0.166 mL) , 1.35 mmol), N- [2- (tert-butyldimethylsiloxy) ethyl] -4- [3- (2,2-dimethylpropionyl) -5- (2,2-dimethylpropionylamino) -2-phenyl -2,3-Dihydro-1,3,4-thiadiazol-2-yl] butanamide (68.0 mg, 83%) was obtained.

Step 8: N- [2- (tert-butyldimethylsiloxy) ethyl] -4- [3- (2,2-dimethylpropionyl) -5- (2,2-dimethylpropionylamino) obtained in Step 7 above 2-Phenyl-2,3-dihydro-1,3,4-thiadiazol-2-yl] butanamide (71.0 mg, 0.117 mmol) was dissolved in THF (1 mL) and 1 mol / L tetrabutylammonium fluoride was dissolved. In THF (0.16 mL) was added and stirred at room temperature for 50 minutes. Water (1 mL) was added to the mixture, and the mixture was extracted with ethyl acetate. The organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by column chromatography (chloroform / methanol = 9/1) to give compound o {4- [3- (2,2-dimethylpropionyl) -5- (2,2-dimethylpropionylamino) -2. -Phenyl-2,3-dihydro-1,3,4-thiadiazol-2-yl] -N- (2-hydroxyethyl) butanamide} (47.6 mg, 85%) was obtained as a white solid.
¹ H NMR (300 MHz, CDCl ₃ ) δ (ppm): 1.28 (s, 9H), 1.33 (s, 9H), 1.56 (m, 1H), 2.22-2.51 (m, 4H), 3.15 (m, 1H ), 3.35 (m, 1H), 3.45 (m, 1H), 3.61-3.76 (m, 2H), 6.31 (br s, 1H), 7.41-7.72 (m, 5H), 8.05 (br s, 1H).
APCI-MS m / z: 477 (M + H) ⁺ .

Claims (11)

Formula (I)

{Wherein n represents an integer of 1 to 3,
R ¹ represents a hydrogen atom,
R ² represents lower alkyl,
Or R ¹ and R ² together represent alkylene,
R ³ represents lower alkyl,
R ⁴ is a hydrogen atom,
NHSO ₂ R ⁶ (wherein R ⁶ is a lower alkyl optionally having 1 to 2 substituents selected from the group consisting of hydroxy, lower alkoxy, amino, lower alkylamino and di-lower alkylamino, Or lower alkenyl)
NHR ⁷ wherein R ⁷ is a lower alkyl optionally having 1 or 2 substituents selected from the group consisting of hydroxy, lower alkoxy, amino, lower alkylamino and di-lower alkylamino, COR ⁸ Wherein R ⁸ is selected from the group consisting of hydroxy, lower alkoxy, amino, lower alkylamino, di-lower alkylamino, carboxy, phenyl, hydroxyphenyl, imidazolyl, guanidyl, methylthio and lower alkoxycarbonylamino Represents a lower alkyl which may have one substituent, a lower alkoxycarbonyl or a nitrogen-containing aliphatic heterocyclic group which may be substituted with oxo, or a lower alkoxy) or a hydrogen atom] or CONHR ⁹ ( wherein, R ⁹ is hydroxy, lower alkoxy, amino, lower alk Arylamino and have 1-2 substituents selected from the group consisting of di-lower alkylamino represents an even lower alkyl) represents,
R ⁵ represents aryl optionally having 1 to 3 substituents selected from the group consisting of halogen, hydroxy, lower alkoxy, nitro, amino, cyano and carboxy}
A therapeutic and / or prophylactic agent for glaucoma comprising the thiadiazoline derivative represented by the formula (I) or a pharmacologically acceptable salt thereof: The following formula (II) in which the specific rotation at 20 ° C. with respect to the sodium D line (wavelength: 589.3 nm) when the thiadiazoline derivative is dissolved in methanol is negative

The therapeutic and / or prophylactic agent according to claim 1, which is a thiadiazoline derivative represented by the formula: wherein R ¹ , R ² , R ³ , R ⁴ , R ⁵ and n are as defined above. The therapeutic and / or prophylactic agent according to claim 1 or 2, wherein R ⁵ is phenyl. The therapeutic and / or prophylactic agent according to any one of claims 1 to 3, wherein R ³ is methyl, ethyl, isopropyl or tert-butyl. Therapeutic and / or prophylactic agent according to any one of claims 1 to 4 R ¹ is a hydrogen atom. The therapeutic and / or prophylactic agent according to any one of claims 1 to 5, wherein R ² is methyl or tert-butyl. Therapeutic and / or prophylactic agent according to claim 1 is a trimethylene or tetramethylene R ¹ and R ² together. The therapeutic and / or prophylactic agent according to any one of claims 1 to 7, wherein R ⁴ is NHSO ₂ R ⁶ (wherein R ⁶ has the same meaning as described above). The therapeutic and / or prophylactic agent according to any one of claims 1 to 7, wherein R ⁴ is CONHR ⁹ (wherein R ⁹ is as defined above). n is 1 or 2, The therapeutic and / or prophylactic agent in any one of Claims 1-9. Thiadiazoline derivatives are represented by the following formulas (a) to (o)

The therapeutic and / or prophylactic agent according to claim 2, which is a thiadiazoline derivative represented by any one of the following: