JPWO2009004899A1 - Sialidase inhibitor - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a novel sialidase inhibitor that inhibits the activity of sialidase existing in Trypanosoma cruzi and the like. An N-acylneuraminic acid derivative having an aromatic ring introduced at the 9-position and 2α-position has a strong sialidase activity inhibitory effect. The present invention is a sialidase inhibitor comprising an N-acylneuraminic acid derivative (the following formula, R1 and R2 each independently represents a substituent having an aromatic ring) or a pharmaceutically acceptable salt thereof. [Chemical 3] [Selected figure] None

Description

  The present invention relates to an N-acylneuraminic acid derivative having sialidase inhibitory activity.

Sialidase (neuraminidase), an enzyme having the ability to cleave N-acetylneuraminic acid known as sialic acid from a sugar chain, is present in many microorganisms and viruses. These neuramidase-producing pathogens are involved in major infectious diseases in humans and other animals.In particular, influenza viruses, Newcastle disease viruses, Vibrio cholerae, trypanosomas, etc. are reported every year, causing enormous damage. Is causing. Accordingly, various neuramidase inhibitors have been developed based on the idea that inhibitors of sialidase activity prevent infection by sialidase-containing viruses and the severity of disease states.
Most of these sialidase inhibitors are analogs and derivatives of 2-deoxy-2,3-dehydro-N-acetylneuraminic acid (DANA), which mimics the original state by mimicking the transition state of the sialic acid cleavage process. It is expected to bind to sialidase more strongly than the sialylated sugar chain as a substrate (Non-patent Document 1). As an analogue or derivative of such 2-deoxy-2,3-dehydro-N-acetylneuraminic acid (DANA), one substituted at the 4-position (Patent Document 1), one substituted at the 3-position (patent Documents 2) and 9-substituted ones (Patent Documents 3 and 4) have been developed as inhibitors of sialidase activity and influenza drugs.
However, some sialidases, such as Trypanosoma cruzi (Non-patent Document 2), are hardly affected by DANA, and therefore, development of inhibitors based on a new basic skeleton is expected.

Japanese Patent No. 2925863 JP 2001-131074 A JP-A-10-139774 Japanese Patent No. 3209946 Virology, 58, 457-463 (1974) J. Mol. Biol., 345, 923-934 (2005)

  The present invention provides a novel sialidase inhibitor that inhibits the activity of sialidase present in trypanosomes and the like.

（式中、Ｒ^１及びＲ^２は、それぞれ独立に、芳香環を有する置換基を表す。）で表される化合物若しくはその製薬上許容される塩又はそれらの溶媒和物に関する。The present inventors have found that introduction of an aromatic ring at positions 9 and 2α of N-acetylneuraminic acid has a strong inhibitory effect on sialidase activity, leading to the completion of the present invention.
That is, the present invention provides a compound of formula (I)

(Wherein R ¹ and R ² each independently represents a substituent having an aromatic ring), or a pharmaceutically acceptable salt thereof, or a solvate thereof.

（式中、Ｒ^３は、置換基を有していてもよい脂肪族基又は置換基を有していてもよい芳香族基を表し、Ｘは単結合、−Ｓ−、−（ＣＨ_２）_ｎ−（式中、ｎは１〜５の自然数を表す。）、−ＣＨ＝ＣＨ−又は−Ｃ≡Ｃ−を表し、Ｒ^４は、置換基を有していてもよい芳香族基又は芳香環を有する脂肪族基を表す。）で表される上記（１）記載の化合物若しくはその製薬上許容される塩又はそれらの溶媒和物。
（３）Ｒ^３が、置換基を有していてもよいアリール基、置換基を有していてもよい脂肪族環基、置換基を有しいてもよいヘテロ芳香族基チオアルキル基、又は置換基を有していてもよいアリールカルバモイルアルキル基である、上記（２）記載の化合物若しくはその製薬上許容される塩又はそれらの溶媒和物。
（４）上記（１）から（３）のいずれかに記載の化合物、若しくはその製薬上許容される塩又はそれらの溶媒和物を有効成分とする医薬組成物。
（５）上記（４）記載の医薬組成物から成るシアリダーゼ阻害剤。Furthermore, the present invention relates to the following (2) to (5).
(2) Formula (II):

(In the formula, R ³ represents an aliphatic group which may have a substituent or an aromatic group which may have a substituent, and X is a single bond, —S—, — (CH ₂ ). _n — (wherein _n represents a natural number of 1 to 5), —CH═CH— or —C≡C—, and R ⁴ represents an aromatic group or aromatic group which may have a substituent. Or a pharmaceutically acceptable salt thereof or a solvate thereof. The compound according to (1) above, which is represented by an aliphatic group having a ring.
(3) R ³ may have an aryl group which may have a substituent, an aliphatic ring group which may have a substituent, a heteroaromatic group thioalkyl group which may have a substituent, or a substituent The compound according to (2) above, which is an arylcarbamoylalkyl group which may have a group, or a pharmaceutically acceptable salt thereof, or a solvate thereof.
(4) A pharmaceutical composition comprising as an active ingredient the compound according to any one of (1) to (3) above, or a pharmaceutically acceptable salt thereof, or a solvate thereof.
(5) A sialidase inhibitor comprising the pharmaceutical composition according to (4) above.

  Examples of viruses and microorganisms that produce the sialidase that is the target of the sialidase inhibitor of the present invention include, for example, parasites such as Trypanosoma cruzi, Trypanosoma brucei, Cryptosporidium and Cryptosporidium perbum, Cholera, Ursius, Gas Examples include bacteria such as gangrene, Streptococcus pneumoniae, Helicobacter pylori and Arthrobacter sialofilus, viruses such as influenza virus, parainfluenza virus, poultry plague virus, mumps virus, Sendai virus and Newcastle disease virus.

A large number of crystal structures of sialidase centering on influenza sialidase have been reported. As its structural feature, the active site has a structure called β-propeller, and seven characteristic amino acids have been found.
When sialic acid and its derivatives approach sialidase, sialidase recognizes the carboxylic acid site of sialic acid at its three arginine residues, and one of the arginines is stabilized by the glutamic acid residue forming a salt pair. Furthermore, it is known that the hydrolytic site requires a hydrogen bond relay structure of tyrosine and glutamic acid and an aspartic acid residue (J. Mol. Biol., 335, 1343-1357 (2004)). .
In the form in which sialic acid or its derivative is bound to sialidase, a loop structure around the sialidase active site protrudes from the sialic acid near the 3rd to 5th acetamide site, and the groove structure of sialidase is an anomer of sialic acid. From the position axial plane (2nd position) to the 9th hydroxyl group side of sialic acid (Structure, 13, 803-815 (2005)). This groove structure is thought to be a space for recognizing the sugar chain part to which sialic acid is bound. Bacteria such as influenza sialidase (Nature, 363, 418-423 (1993)) and Vibrio cholerae Secretory sialidase produced (J. Biol. Chem., 279, 40819-40826 (2004)), sialidase produced by mammals such as humans (J. Biol. Chem., 280, 469-475 (2005)), etc. It exists as a common structure.

The sialidase inhibitor of the present invention focuses on the groove structure of the sialidase active site, and obtains a specific and strong inhibitory ability by substituting the 9-position hydroxyl group and 2-position α side corresponding to the groove structure. There is a feature. Moreover, the groove | channel corresponding to the 9th-position from the 2nd alpha side of the binding sialic acid (sialidase inhibitor) is a characteristic structure of sialidase in general.
As such a substituent, the aromatic ring has a planar structure similar to the pyranose ring or furanose ring of the sugar, and has a binding force to the groove structure around the sialidase active site that recognizes the sugar chain bound to sialic acid. (Structure, 12, 775-784 (2004)). The N-acylneuraminic acid derivative of the present invention must have aromatic rings at the 9-position and the 2α-position. When aromatic rings are introduced at both the 9-position and the 2α-position, the spatial positions of sialic acid and sialidase can be controlled by π-π stacking, so that sialic acid can be efficiently incorporated into the target sialidase groove structure. It becomes possible to adapt to.
Since the N-acylneuraminic acid derivative of the present invention has such a substituent, it inhibits the activity of sialidase by a mechanism that has not functioned in DANA and its derivatives that have been used as the basic skeleton of sialidase inhibitors so far. It is thought that it will be possible.
In addition, since a large amount of esterase (lipase) is present in cells or in vivo, the substituents at the 9-position and 2α-position are considered under such conditions when actual use as a sialidase inhibitor is considered. Therefore, it is required not to be easily decomposed. For example, since ester groups (Patent Documents 2 and 4) are rapidly decomposed, it is considered that there is no substitution effect in actual use.

式中、Ｒ^１及びＲ^２は、それぞれ独立に、芳香環を有する置換基を表す。この置換基は芳香環を、好ましくは１〜５個、より好ましくは１〜３個有する。
このＲ^１及びＲ^２は、０〜５原子（主鎖上の原子数として）から成る結合基を介して、９位又は２位の炭素に結合する芳香環を有することが好ましい。芳香環が５原子以上から成る結合基を介して９位又は２位の炭素に結合する場合には、この芳香環は、シアル酸（シアリダーゼ阻害剤）がシアリダーゼ活性部位周辺の溝構造に結合する際に十分な結合力を示さない場合もあると考えられる。また０原子から成る結合基を介して９位又は２位の炭素に結合するということは、芳香環が９位又は２位の炭素に直接結合することを意味する。
この芳香環としては、ベンゼン環、ナフタレン環、アントラセン環、ピレン環などの炭化水素芳香族環が挙げられるが、ピリジン環、アクリジン環、キノリン環、ピリダジン環、ピリミジン環、ピラジン環、ピロール環、ピラゾール環、トリアゾール環、フラン環、チオフェン環等のヘテロ芳香環をも含む。これらの芳香環は如何なる形態や結合数で置換基に結合されていてもよい。例えば、ベンゼン環は、フェニル基やフェニレン基などとして置換基中に存在してもよい。That is, the sialidase inhibitor of the present invention comprises an N-acylneuraminic acid derivative having a substituent having an aromatic ring at the 9-position and the 2α-position, and is represented by the following structural formula.

In the formula, R ¹ and R ² each independently represent a substituent having an aromatic ring. This substituent preferably has 1 to 5 aromatic rings, more preferably 1 to 3 aromatic rings.
R ¹ and R ² preferably have an aromatic ring bonded to the 9th or 2nd carbon via a bonding group consisting of 0 to 5 atoms (as the number of atoms on the main chain). When the aromatic ring is bonded to the 9th or 2nd carbon via a linking group consisting of 5 or more atoms, this aromatic ring binds sialic acid (sialidase inhibitor) to the groove structure around the sialidase active site. In some cases, sufficient bonding strength may not be exhibited. In addition, bonding to the 9th or 2nd carbon via a bonding group consisting of 0 atoms means that the aromatic ring is directly bonded to the 9th or 2nd carbon.
Examples of the aromatic ring include hydrocarbon aromatic rings such as benzene ring, naphthalene ring, anthracene ring, pyrene ring, pyridine ring, acridine ring, quinoline ring, pyridazine ring, pyrimidine ring, pyrazine ring, pyrrole ring, Heteroaromatic rings such as a pyrazole ring, a triazole ring, a furan ring and a thiophene ring are also included. These aromatic rings may be bonded to the substituent in any form or number of bonds. For example, the benzene ring may be present in the substituent as a phenyl group or a phenylene group.

Any method may be used to bond the aromatic ring to the 9-position and the 2α-position. Examples of such bonding methods include olefin metathesis method, Suzuki-Kajiura cross-coupling method, Heck reaction, Michael addition reaction, aldol type reaction, direct introduction of aromatic ring by Sn2 type or Sn1 type substitution reaction, carbon-carbon bond ( -C-, -C = C-) formation, dehydration condensation reaction, amide (-NHCO-), urea (-NHCONH-), imide (-N (CO-) CO-), sulfone using activated ester Formation of acid amide (—NHSO ₂ —) bond, ether (—O—), thioether (—S—), selenoether (—Se—), amine (—NH—, —N) by Sn2 type or Sn1 type substitution reaction Formation of (−)-) · alkyne (—C≡C—) bond, acetal type bond such as acetal (—C (—O—) O—), thioacetal (—C (—S—) S—) form , Imine (—N═C—, —C═N—), oxime (—O—N═C—, —C═N—O—), hydrazide (—NH—N═C—, —C═N—) NH-) and other imine-type bond formation, azide-mediated bond via a triazole ring, Diels-Alder reaction, and ring formation reaction by tandem reaction combining the above-mentioned reactions.
These linking groups may be present between a plurality of aromatic rings when a plurality of aromatic rings are introduced.

In this, as a general method, N- 9-position hydroxyl group of the azide of acyl neuraminic acid ^{(-N = N + = N -} ) is converted to group (Tetrahedron, 50, 7445-7460 (1994 )), or N- acylneuraminic An azide (—N═N ⁺ ═N ⁻ ) group was introduced into the 2α position of the acid by the SN2 reaction (Carbohydrate Research, 291, 183-187 (1996)), while the substituent to be introduced was acetylene (—C≡CH ) Is bonded, and these are reacted in the presence of a copper catalyst to introduce a desired substituent into the 9-position or 2α-position of N-acylneuraminic acid via a triazole ring (formula below). be able to. This triazole ring is not an essential element as a sialidase inhibitor of the present invention, but is inevitably introduced to introduce a desired substituent.

  Further, the substituent having an aromatic ring further has a halogen atom, a hydroxyl group, an alkyl group having 1 to 4 carbon atoms, a formyl group, an amino group, a nitro group, a cyano group, a carboxyl group, a sulfonic acid group, and a carbon number. Substitution wherein the alkyl group 1 to 4 is selected from a group (substituent group A) substituted with a halogen atom, a hydroxyl group, a formyl group, an amino group, a nitro group, a cyano group, a carboxyl group, a sulfonic acid group, or the like It may be substituted with a group.

Any of the above-mentioned linking groups can be used to bond the aromatic ring to the 9-position, but from the viewpoint of stability under physiological conditions, the linking group (Y) for bonding the aromatic ring to the 9-position is preferably Is a single bond, a divalent triazole ring (Chemical Formula 4), -O-, -S-, -Se-, -NHCO-, -NH-, -N (-)-, -NHCONH-, -N (CO- ) CO—, — (CH ₂ ) _n — (wherein n represents a natural number of 1 to 5), —CH═CH—, or —C≡C—.
In this case, R ¹ is represented as -YR ³ .

R ³ represents an aromatic group which may have a substituent or an aliphatic group having an aromatic ring. However, when Y is a divalent triazole ring (Formula 4), R ³ may be an aliphatic group having no aromatic ring.
Examples of this aromatic group include an aryl group and a heteroaromatic group. The aryl group is preferably a phenyl group or an α- or β-naphthyl group. Examples of the heteroaromatic group include residues of the above heteroaromatic ring, for example, furyl group, thienyl group, pyrrolyl group, imidazolyl group, pyrazolyl group, thiazolyl group, isothiazolyl group, oxazolyl group, isoxazolyl group, triazolyl group, oxadiazolyl group, Examples include thiadiazolyl group, tetrazolyl group, pyridyl group, pyrimidinyl group, pyridazinyl group, pyrazinyl group and the like.
This aromatic ring has the same definition as above.
Examples of the aliphatic group include a linear or branched alkyl group, an aliphatic ring group such as a cyclohexane ring, or a combination thereof.
This aliphatic group preferably has 1 to 20 carbon atoms, more preferably 1 to 12 carbon atoms.
The aliphatic group having an aromatic ring preferably has 1 to 5 aromatic rings, more preferably 1 to 3 aromatic rings.
Further, these aromatic groups, aliphatic groups having an aromatic ring, and aliphatic groups not having an aromatic ring may further have the above-described substituent.

Further, R ³ represents an aryl group which may have a substituent, an aliphatic ring group which may have a substituent, a heteroaromatic group thioalkyl group which may have a substituent, or a substituent. An arylcarbamoylalkyl group which may have a hydrogen atom is preferable.
The substituent of the aryl group which may have a substituent in R ³ is selected from an arylcarbamoylcarbamoyl group which may be substituted with a substituent selected from Substituent Group A, and Substituent Group A. An arylcarbamoyl group optionally substituted with a substituent.
Examples of the substituent of the aliphatic ring group which may have a substituent in R ³ include an aryloxy group and a hydroxy group which may be substituted with a substituent selected from the substituent group A. .
Examples of the substituent of the heteroaromatic thioalkyl group which may have a substituent in R ³ include an aryl group which may be substituted with a substituent selected from the substituent group A.
Examples of the substituent of the arylcarbamoylalkyl group which may have a substituent in R ³ include an aryl group which may be substituted with a substituent selected from the substituent group A, a cycloalkyl group, and the like. .

On the other hand, the linking group for bonding the aromatic ring to the 2α-position is chemically and biologically stable compared to the linking group in the 9-position because the oxygen atom is bonded to the 2-position carbon of N-acylneuraminic acid. Restricted for sex reasons. That is, the linking group (X) for bonding the aromatic ring to the 2α position is preferably a single bond, a divalent triazole ring (Chemical Formula 4), —O—, —S—, —Se—, —NHCO—, — (CH ₂ ) _n — (wherein _n represents a natural number of 1 to 5), —CH═CH—, —C≡C—, more preferably a single bond, —S—, — (CH ₂ ) _n- (wherein _n represents a natural number of 1 to 5), -CH = CH-, or -C≡C-.
In this case, ^{R 2} is represented as ^{-X-R 4.} In this formula, R ⁴ is defined similarly to R ³ .

In the present specification, the “solvate” includes, for example, solvates with organic solvents, hydrates and the like. When forming a hydrate, it may be coordinated with any number of water molecules.
The compounds represented by formula (I) and formula (II) include pharmaceutically acceptable salts. For example, alkali metals (such as lithium, sodium or potassium), alkaline earth metals (such as magnesium or calcium), ammonium, salts with organic bases and amino acids, or inorganic acids (hydrochloric acid, sulfuric acid, nitric acid, hydrobromic acid, phosphorus Acid or hydroiodic acid) and organic acids (acetic acid, trifluoroacetic acid, citric acid, lactic acid, tartaric acid, oxalic acid, maleic acid, fumaric acid, mandelic acid, glutaric acid, malic acid, benzoic acid, phthalic acid, Benzenesulfonic acid, p-toluenesulfonic acid, methanesulfonic acid, ethanesulfonic acid and the like). Particularly preferred are hydrochloric acid, phosphoric acid, tartaric acid, methanesulfonic acid and the like. These salts can be formed by a commonly performed method.

  The pharmaceutically acceptable salt of the N-acylneuraminic acid derivative of the present invention is preferably a salt formed by ionizing the carboxylic acid at the 1-position, and may be, for example, an inorganic base or an organic base and a salt. Preferred salts include, for example, salts with inorganic bases such as sodium, potassium, magnesium, calcium and aluminum, salts with organic bases such as methylamine, ethylamine, diethylamine, triethylamine, tetraethylammonium, guanidine and ethanolamine, lysine, Examples thereof include salts with basic amino acids such as arginine and ornithine.

Further, the compounds represented by formula (I) and formula (II) are not limited to specific isomers, but all possible isomers (keto-enol isomer, imine-enamine isomer, diastereoisomerism) Isomers, optical isomers, rotational isomers, etc.) and racemates.
When the compound according to the present invention is administered to humans, it can be administered orally as powders, granules, tablets, capsules, pills, liquids, etc., or non-injectables, suppositories, transdermal absorption agents, inhalants, etc. It can be administered orally. In addition, excipients, binders, wetting agents, disintegrants, lubricants and the like suitable for the dosage form may be mixed with an effective amount of this compound as necessary to obtain a pharmaceutical preparation. it can.
The dose varies depending on the disease state, administration route, patient age, or body weight, but is usually 0.1 μg to 1 g / day, preferably 0.01 to 200 mg / day when orally administered to an adult. In the case of parenteral administration, it is usually 1 μg to 10 g / day, preferably 0.1 to 2 g / day.

Furthermore, the sialidase inhibitor of the present invention includes pharmacologically acceptable prodrugs. For example, a group that can be converted into a hydroxyl group, a 1-position carboxylic acid, or a 5-position acetamide group by hydrolysis, solvolysis or under physiological conditions. For example, as a prodrug of a hydroxyl group, -OCO-substitution may be performed. Lower alkylene-COOR ⁵ (R ⁵ represents a hydrogen atom or lower alkyl; the same shall apply hereinafter), —OCO—optionally substituted lower alkenylene—COOR ⁵ , —OCO—optionally substituted aryl, —OCO - lower alkylene -O- lower alkylene ^{-COOR 5, -OCO-COR 5,} -OCO- optionally substituted lower alkyl, -OSO ₂ - optionally substituted lower alkylene -COOR ⁵ also, -O- Futajiru, 5 -Methyl-1,3-dioxolen-2-one-4-yl-methyloxy and the like, and as a prodrug of carboxylic acid Examples thereof include esterification with an optionally substituted lower alkylene group and conversion reaction to an aldehyde group or acetal group, and examples of the prodrug of an acetamide group include conversion to an imide.

The following examples illustrate the invention but are not intended to limit the invention.
Synthesis example 1
According to the synthetic scheme shown in FIG. 1, 5-acetylneuraminic acid (Neu5Ac, Nacalai Tesque, Inc.) to 5-acetamide-9-azido-3,5,9-trideoxy-2-S- (4-nitrophenyl)- D-glyceo-α-D-galacto-non-2-uropyranosonic acid (Compound 1) was synthesized.
5-acetylneuraminic acid (Neu5Ac, Nacalai Tesque Co., Ltd.) (2.57 g) suspended in methanol (37 ml) and methanol-washed acid ion exchange resin Dowex (R) 50W (Dow Chemical Co.) (2.5 g) ) And stirred vigorously overnight. After the resin was filtered off, the filtrate was concentrated, acetyl chloride (23 ml) was added to the residue, and the mixture was vigorously stirred for 2 days. The residue and 4-nitrothiophenol (3.45 g) were dissolved in dry acetonitrile (73 ml), and ethyldiisopropylamine (3.6 ml) was added dropwise with stirring. After stirring at room temperature overnight, the reaction mixture was concentrated, the residue was purified by silica gel column chromatography [ethyl acetate], and methyl 5-acetamide-4,7,8,9-O-tetraacetyl-3,5- Dideoxy-2-O-methyl-2-S- (4-nitrophenyl) -D-glyceo-α-D-galacto-non-2-uropyranosonate (compound 2) (2.9 g, 56%). Obtained. Compound 2: (TLC) Rf 0.3 [ethyl acetate].

  Next, 28% sodium methylate solution (0.45 ml) was added to a dry methanol (45 ml) solution of compound 2 (2.9 g) obtained above, and the mixture was stirred at room temperature for 1 hour. Crystals precipitated during the reaction were collected by filtration and methyl-5-acetamido-3,5-dideoxy-2-S- (4-nitrophenyl) D-glyceo-α-D-galacto-non-2-uropyranosonate. (Compound 3) (1.12 g, 52%) was obtained. Compound 3: (TLC) Rf 0.5 [chloroform-methanol (4: 1)].

  Next, tosyl chloride (0.84 g) and DMAP (30 mg) were sequentially added to a pyridine (27 ml) solution of Compound 3 (1.1 g) obtained above, and the mixture was stirred overnight at room temperature. The reaction mixture was concentrated, and the residue was purified by silica gel column chromatography [chloroform-methanol (9: 1)] to give methyl-5-acetamido-3,5-dideoxy-2-S- (4-nitrophenyl) -9. -O-tosyl-D-glyceo-α-D-galacto-non-2-uropyranosonate (compound 4) (1.3 g, 87%) was obtained. Compound 4: (TLC) Rf 0.5 [chloroform-methanol (8: 1)].

Next, sodium azide (0.55 g) was added to a DMF (23 ml) solution of compound 4 (1.3 g) obtained above, and the mixture was stirred at 100 ° C. overnight and concentrated. The residue was purified by silica gel column chromatography [chloroform-methanol (2: 1)] to give 5-acetamide-9-azido-3,5,9-trideoxy-2-S- (4-nitrophenyl) -D-glyceo. -Α-D-galacto-non-2-uropyranosonic acid (Compound 1) (0.51 g, 51%) was obtained. Compound 1: (TLC) Rf 0.4 [chloroform-methanol (2: 1)].
The analysis value of the synthesized compound 1 is shown below.
¹ H NMR (600 MHz, D ₂ O) δ 8.29-7.82 (m, 2H; aromatic), 3.91 (m, 2H; H-8, H-5), 3.74 (ddd, 1H, J = 4.2, 10.2, 11.4 Hz; H-4), 3.67 (dd, 1H, J = 1.8, 13.2 Hz; H-7), 3.59 (m, 2H; H-6, H-9a), 3.43 (q, 1H, J = 6.6 , 13.2 Hz; H-9b), 2.97 (dd, 1H, J = 4.2, 12.6 Hz; H-3eq), 2.08 (s, 3H; Ac), 1.95 (dd, 1H, J = 11.4, 12.6 Hz; H -3ax); ESI-MS Calcd for C ₁₇ H ₂₁ N ₅ O ₉ S [MH]-470.44174, found 470.22825.

Synthesis example 2
Tris- (1-Benzyltriazol-4-ylmethyl) amine (Sigma Aldrich) (5 mM, DMSO solution, 100 μL), copper sulfate (5 mM, H ₂ O : t-BuOH = 1: 1 solution, 100 μL), sodium ascorbate (50 mM, H ₂ O: t-BuOH = 1: 1 solution, 100 μL), Compound 1 obtained in Synthesis Example 1 (100 mM, H ₂ O: t-BuOH = 1: 1 solution, 100 μL), 2,6-dichloro-N- (3-ethynylphenylcarbonyl) benzamide (2,6-dichloro-N- (3-ethynylphenylcarbamoyl) benzamide, Maybridge) (100 mM, H ₂ O: t-BuOH = 1: 1 solution, 100 μL) were sequentially mixed and stirred at room temperature for 8 hours. The progress of the reaction was confirmed by ESI-MS, and the reaction solution was directly purified by reversed-phase HPLC to obtain 5-acetamido-9- (4- (3- (3- (2,6-dichlorobenzoyl) ureido) phenyl). -1H-1,2,3-triazol-1-yl) -3,5,9-trideoxy-2- (4-nitrophenylthio) -D-glyceo-α-D-galacto-non-2-uropyranosonic acid (Compound 5, the following formula) was obtained (yield 3 mg, yield 37%).

The analysis value of the synthesized compound 5 is shown below.
¹ H NMR (600 MHz, CD ₃ OD) δ 8.24-7.43 (m, 11H; Aromatic), 4.83 (m, 1H; H-9a), 4.44 (dd, 1H, J = 8.4, 14.4 Hz; H-9b ), 4.09 (ddd, 1H, J = 7.2, 8.4; H-8), 3.81 (d, 1H, J = 10.2 Hz; H-5), 3.69 (ddd, 1H, J = 4.8, 10.2, 10.8 Hz; H-4), 3.43 (d, 1H, J = 10.2 Hz; H-6), 3.38 (d, 1H, J = 8.4 Hz; H-7), 2.87 (dd, 1H, J = 4.8, 12.6 Hz; H-3eq), 1.95 (s, 3H; Ac), 1.89 (dd, 1H, J = 11.4, 12.0 Hz; H-3ax); ESI-MS Calcd for C ₃₃ H ₃₁ C _l2 N ₇ O ₁₁ S (MH ]-802.11793, found 802.00916.

Synthesis example 3
Tris- (1-Benzyltriazol-4-ylmethyl) amine (Sigma Aldrich) (5 mM, DMSO solution, 100 μL), copper sulfate (5 mM, H ₂ O : t-BuOH = 1: 1 solution, 100 μL), sodium ascorbate (50 mM, H ₂ O: t-BuOH = 1: 1 solution, 100 μL), Compound 1 obtained in Synthesis Example 1 (100 mM, H ₂ O: t-BuOH = 1: 1 solution, 100 μL), 17-α-ethynyl-3- (4-nitrophenoxy) -1,3,5 (10) -estraditrien-17β-ol (17α-ethynyl- 3- (4-nitrophenoxy) -1,3,5 (10) -estratrien-17β-ol, Maybridge) (100 mM, H ₂ O: t-BuOH = 1: 1 solution, 100 μL) And stirred at room temperature for 8 hours. The progress of the reaction was confirmed by ESI-MS, the reaction solution was directly purified by reverse phase HPLC, and 5-acetamide-3,5,9-trideoxy-9- (4- (17β-hydroxy-3- (4- Nitrophenoxy) -1,3,5 (10) -estraditri-17α-yl) -1H-1,2,3-triazol-1-yl) -2-S- (4-nitrophenyl) -D-glyceo -Α-D-galacto-non-2-uropyranosonic acid (compound 6, the following formula) was obtained (yield 5 mg, yield 56%).

Analytical values of the synthesized compound 6 are shown below.
¹ H NMR (600 MHz, CD ₃ OD) δ 8.18-6.78 (m, 12H; Aromatic), 4.73 (dd, 1H, J = 3.0, 14.4 Hz; H-9a), 4.43 (dd, 1H, J = 7.8 , 14.4 Hz; H-9b), 4.15 (ddd, 1H, J = 3.0, 7.8, 9.0 Hz; H-8), 3.79 (t, 1H, J = 10.2 Hz; H-5), 3.68 (ddd, 1H , J = 4.8, 10.2, 10.8 Hz; H-4), 3.46 (d. 1H, J = 10.2 Hz; H-6), 3.38 (d, 1H, J = 9.0 Hz; H-7), 2.87 (dd , 1H, J = 4.8, 12.6 Hz; H-3eq), 2.83 (m, 2H), 2.44 (m, 1H), 2.22-1.99 (m, 4H), 1.97 (s, 3H; Ac), 1.87 (t , 1H, J = 12.0 Hz; H-3ax), 1.76-1.38 (m, 6H), 1.25 (m, 1H), 1.05 (s, 3H), 0.76 (m, 1H); ESI-MS Calcd for C ₄₃ H ₄₈ N ₆ O ₁₃ S [MH]-887.30001, found 887.21774.

Synthesis example 4
Tris- (1-Benzyltriazol-4-ylmethyl) amine (Sigma Aldrich) (5 mM, DMSO solution, 100 μL), copper sulfate (5 mM, H ₂ O : t-BuOH = 1: 1 solution, 100 μL), sodium ascorbate (50 mM, H ₂ O: t-BuOH = 1: 1 solution, 100 μL), Compound 1 obtained in Synthesis Example 1 (100 mM, H ₂ O: t-BuOH = 1: 1 solution, 100 μL), 3- (4-tert-butylphenyl) -5- (2-propynylthio) -4- (3- (trifluoromethyl) phenyl) -4H— 1,2,4-triazole (3- (4-tert-butylphenyl) -5- (prop-2-ynylthio) -4- (3- (trifluoromethyl) phenyl) -4H-1,2,4-triazole, Maybridge (100 mM, H ₂ O: t-BuOH = 1: 1 solution, 100 μL) were sequentially mixed and stirred at room temperature for 8 hours. The progress of the reaction was confirmed by ESI-MS, the reaction solution was directly purified by reverse phase HPLC, and 5-acetamido-3,5,9-trideoxy-9- (4-((5- (4-tertiary butyl Phenyl) -4- (3- (trifluoromethyl) phenyl) -4H-1,2,4-triazol-3-ylthio) methyl) -1H-1,2,3-triazol-1-yl) -2- S- (4-nitrophenyl) -D-glyceo-α-D-galacto-non-2-uropyranosonic acid (compound 7, formula below) was obtained (yield 9 mg, 100%).

Analytical values of the synthesized compound 7 are shown below.
¹ H NMR (600 MHz, CD ₃ OD) δ 7.97-7.27 (m, 13H, Aromatic), 4.59 (m, 1H; H-9a), 4.48 (s, 2H; CH2), 4.24 (dd, 1H, J = 8.4, 13.8 Hz; H-9b), 3.94 (t, 1H, J = 8.4 Hz; H-8), 3.79 (t, 1H, J = 10.2 Hz; H-5), 3.67 (ddd, 1H, J = 4.8, 10.2, 10.8 Hz; H-4), 3.39 (d, 1H, J = 10.8 Hz; H-6), 3.35 (d, 1H, J = 9.0 Hz; H-7), 2.86 (dd, 1H , J = 4.8, 12.6 Hz; H-3eq), 1.95 (s, 3H; Ac), 1.88 (dd, 1H, J = 11.4, 12.6 Hz; H-3ax), 1.25 (s, 9H; CH3 × 3) ; ESI-MS Calcd for C ₃₉ H ₄₁ F ₃ N ₈ O ₉ S ₂ [MH]-885.23900, found 885.17536.

Synthesis example 5
Tris- (1-benzyltriazol-4-ylmethyl) amine (12.5 mM, DMSO solution, 100 μL), copper sulfate (12.5 mM, H ₂ O: t-BuOH = 1: 1 solution, 100 μL), sodium ascorbate ( 12.5 mM, H ₂ O: t-BuOH = 1: 1 solution, 100 μL), compound 1 obtained in Synthesis Example 1 (250 mM, H ₂ O: t-BuOH = 1: 1 solution, 100 μL), (2- (2-propynylthio) -6- (thiophen-2-yl) -4- (trifluoromethyl) nicotinonitrile (2- (prop-2-ynylthio) -6- (thiophen-2-yl) -4- (trifluoromethyl) nicotinonitrile, Maybridge) (250 mM, H ₂ O: t-BuOH = 1: 1 solution, 100 μL) were mixed in order and stirred at room temperature for 8 hours.The progress of the reaction was confirmed by ESI-MS. The reaction solution was directly purified by reverse phase HPLC, and 5-acetamido-9- (S- (3-cyano-6- (thiophen-2-yl) -4-trifluoromethylpyridin-2-yl))- 1H-1,2,3-Triazol -1-yl) -3,5,9-trideoxy-2-S- (4-nitrophenyl) -D-glyceo-α-D-galacto-non-2-uropyranosonic acid (compound 8, formula below) (Yield 16 mg, 81% yield).

Analytical values of the synthesized compound 8 are shown below.
¹ H NMR (600 MHz, CD ₃ OD) δ 7.93-7.12 (m, 9H; aromatic), 4.58 (m, 3H; H-9a, CH2), 4.12 (dd, 1H, J = 9.6, 13.8 Hz; H -9b), 3.80 (t, 1H, J = 9.6 Hz; H-8), 3.70 (t, 1H, J = 10.2 Hz; H-5), 3.57 (ddd, 1H, J = 4.8, 10.8 Hz; H -4), 3.37 (m, 2H; H-6, H-7), 2.80 (dd, 1H, J = 4.8, 12.0 Hz; H-3eq), 1.90 (s, 3H; Ac), 1.74 (dd, 1H, J = 11.4, 12.0 Hz; H-3ax); ESI-MS Calcd for C ₃₁ H ₂₈ F ₃ N ₇ O ₉ S ₃ [MH]-794.10627, found 794.23757.

Synthesis Example 6
Tris- (1-benzyltriazol-4-ylmethyl) amine (12.5 mM, DMSO solution, 100 μL), copper sulfate (12.5 mM, H ₂ O: t-BuOH = 1: 1 solution, 100 μL), sodium ascorbate ( 12.5 mM, H ₂ O: t-BuOH = 1: 1 solution, 100 μL), compound 1 obtained in Synthesis Example 1 (250 mM, H ₂ O: t-BuOH = 1: 1 solution, 100 μL), 4′− Ethyl-N- (1-ethynylcyclohexyl) bipyridyl-4-carboxamide (4'-ethyl-N- (1-ethynylcyclohexyl) biphenyl-4-carboxamide, Maybridge) (250 mM, H ₂ O: t-BuOH = 1 : 1 solution, 100 μL) were sequentially mixed and stirred at room temperature for 8 hours. The progress of the reaction was confirmed by ESI-MS, the reaction solution was directly purified by reverse phase HPLC, and 5-acetamido-3,5,9-trideoxy-9- (4- (1- (4′-ethylbiphenyl- 4-ylcarboxamido) cyclohexyl) -1H-1,2,3-triazol-1-yl) -2-S- (4-nitrophenyl) -D-glyceo-α-D-galacto-non-2-uropyranosonic acid (Compound 9, the following formula) was obtained (yield 13 mg, yield 65%).

Analytical values of the synthesized compound 9 are shown below.
¹ H NMR (600 MHz, CD ₃ OD) δ 8.09-7.29 (m, 13H; aromatic), 4.78 (dd, 1H, J = 2.4, 14.4 Hz; H-9a), 4.37 (dd, 1H, J = 8.4 , 14.4 Hz; H-9b), 4.13 (ddd, 1H, J = 1.8, 2.4, 9.0 Hz; H-8), 3.83 (t, 1H, J = 10.2 Hz; H-5), 3.71 (ddd, 1H , J = 4.8, 10.2, 10.8 Hz; H-4), 3.52 (dd, 1H, J = 1.2, 10.2 Hz; H-6), 3.38 (dd, 1H, J = 1.2, 9.0 Hz; H-7) , 2.89 (dd, 1H, J = 4.8, 12.6 Hz; H-3eq), 2.69 (dd, 2H, J = 7.8 Hz; CH2), 2.60 (m, 2H), 2.10 (m, 2H), 1.97 (s , 3H; Ac), 1.90 (dd, 1H, J = 11.4, 12.6 Hz; H-3ax), 1.73-1.49 (m, 6H), 1.26 (t, 1H, J = 7.2 Hz; CH3); ESI-MS Calcd for C ₄₀ H ₄₆ N ₆ O ₁₀ S [MH] -801.29961, found 801.40626.

Synthesis example 7
Tris- (1-benzyltriazol-4-ylmethyl) amine (12.5 mM, DMSO solution, 100 μL), copper sulfate (12.5 mM, H ₂ O: t-BuOH = 1: 1 solution, 100 μL), sodium ascorbate ( 12.5 mM, H ₂ O: t-BuOH = 1: 1 solution, 100 μL), Compound 1 obtained in Synthesis Example 1 (250 mM, H ₂ O: t-BuOH = 1: 1 solution, 100 μL), N- ( 4-ethynylphenyl) -3,5-bis (trifluoromethyl) benzamide (N- (4-ethynylphenyl) -3,5-bis (trifluoromethyl) benzamide, Maybridge) (250 mM, H ₂ O: t-BuOH = 1: 1 solution, 100 μL) were sequentially mixed and stirred at room temperature for 8 hours. The progress of the reaction was confirmed by ESI-MS, the reaction solution was directly purified by reverse phase HPLC, and 5-acetamido-3,5,9-trideoxy-9- (4- (4- (3,4-bis ( Trifluoromethyl) benzamido) phenyl) -1H-1,2,3-triazol-1-yl) -2-S- (4-nitrophenyl) -D-glyceo-α-D-galacto-non-2-uropyranosone An acid (compound 10, the following formula) was obtained (yield 14 mg, yield 68%).

Analytical values of the synthesized compound 10 are shown below.
¹ H NMR (600 MHz, CD ₃ OD) δ 8.58-7.80 (m, 10H, aromatic), 4.82 (m, 1H; H-9a), 4.45 (dd, 1H, J = 7.8, 14.4 Hz; H-9b ), 4.12 (dd, 1H, J = 7.8, 9.0 Hz; H-8), 3.83 (dd, 1H, J = 7.2, 9.6 Hz; H-5), 3.71 (m, 1H; H-4), 3.45 (d, 1H, J = 10.2 Hz; H-6), 3.41 (d, 1H, J = 8.4 Hz; H-7), 2.90 (dd, 1H, J = 4.8, 12.6 Hz; H-3eq), 1.98 (s, 3H; Ac), 1.91 (t, 1H, J = 12.6 Hz; H-3ax); ESI-MS Calcd for C ₃₁ H ₂₈ F ₆ N ₆ O ₁₀ S [MH]-827.16483, found 827.27480.

Synthesis example 8
Compound 3 (20 mg) obtained in Synthesis Example 1 was dissolved in 0.1N sodium hydroxide (0.5 mL), and the solution was permeated and stirred for 1 hour. The reaction solution was neutralized with dry ice, concentrated, desalted with Sephadex G-10 (Pharmacia), and S-4-nitrophenyl N-acetyl-α-neuraminic acid (Compound 11) was sodium salt. Got as. (Yield 20 mg, 99% yield).

Analytical values of the synthesized compound 12 are shown below.
¹ H NMR (600 MHz, D ₂ O) δ 8.23 (m, 2H), 7.84 (m, 2H), 3.89 (m, 2H), 3.76 (ddd, 1H, J = 4.8, 10.2, 11.4 Hz), 3.67 (m, 1H), 3.62 (dd, 1H, J = 6.0, 11.4 Hz), 3.47 (m, 2H), 2.97 (dd, 1H, J = 4.8, 12.6 Hz), 2.08 (s, 3H), 1.95 ( (dd, 1H, J = 11.4, 12.6 Hz)

Synthesis Example 9
Tris- (1-benzyltriazol-4-ylmethyl) amine (12.5 mM, DMSO solution, 100 μL), copper sulfate (12.5 mM, H ₂ O: t-BuOH = 1: 1 solution, 100 μL), sodium ascorbate ( 12.5 mM, H ₂ O: t-BuOH = 1: 1 solution, 100 μL), 5-acetamido-9-azido-3,5,9-trideoxy-2-O-methyl-D-glyceo-α-D-galacto Non-2-uropyranosonic acid (prepared according to the literature (Tetrahedron, 50, 7445-7460 (1994))) (250 mM, H ₂ O: t-BuOH = 1: 1 solution, 100 μL), N- (4 -Ethynylphenyl) -3,5-bis (trifluoromethyl) benzamide (N- (4-ethynylphenyl) -3,5-bis (trifluoromethyl) benzamide, Maybridge) (250 mM, H ₂ O: t-BuOH = 1: 1 solution, 100 μL) were sequentially mixed and stirred at room temperature for 8 hours. The progress of the reaction was confirmed by ESI-MS, the reaction solution was directly purified by reverse phase HPLC, and 5-acetamido-3,5,9-trideoxy-9- (4- (4- (3,4-bis ( Trifluoromethyl) benzamido) phenyl) -1H-1,2,3-triazol-1-yl) -2-S- (4-nitrophenyl) -D-glyceo-α-D-galacto-non-2-uropyranosone An acid (compound 12, the following formula) was obtained (yield 15 mg, yield 85%).

Examples 1-6
Trypanosoma cruzi-derived sialidase inhibitory activity was measured for compounds 5-10 obtained in Synthesis Examples 2-7.
In this test, trypanosoma cruzi-derived sialidase (obtained from Prof. Jose Osvaldo Previato of Biofisica Carlos Chagas Filho Laboratory, literature (J. Biol. Chem., 277, 45962-45968 (2002)) And 4-methyl-umbelliferyl 5-acetylneuraminic acid (4-MU NANA, Calbiochem) was used as a substrate.
Sialidase inhibitory activity was carried out by the following procedure. Sodium acetate buffer at pH 5 prepared to a final concentration of 100 mM in advance, enzyme prepared so that the final enzyme activity is 0.0015 units (1 unit is the amount of enzyme that hydrolyzes 1 μmol of sialyl lactose per minute). A solution, an aqueous solution of a test compound prepared to have a final concentration of 10 μM, 30 μM, 50 μM, 100 μM, 300 μM, and 500 μM, and a 4-MU NANA aqueous solution prepared to have a final concentration of 3 mM or 1 mM are sequentially mixed. After stirring for 30 minutes, the reaction was stopped by adding 5 volumes of 100 mM glycine buffer (pH 11), and the excitation wavelength was 365 nm and the fluorescence wavelength was 450 nm using a microplate reader (SpectraMax M5, Molecular Device). The inhibition constant Ki was measured by the Dixon method (Biochem. J., 55, 170-171 (1953)).

Comparative Examples 1-2
For comparison, the compounds 11 to 12 obtained in Synthesis Examples 8 to 9 were examined for sialidase inhibitory activity in the same manner as in Example 1.

本願発明のシアリダーゼ阻害剤は、顕著にトリパノソ−マ・クルジ由来シアリダーゼの活性を阻害した。これは、シアル酸の９位及び２α位の本願発明の置換基が、ノイラミニダーゼの活性阻害に極めて有効であったことを示している。The results are shown in the table below.

The sialidase inhibitor of the present invention remarkably inhibited the activity of Trypanosoma cruzi-derived sialidase. This indicates that the substituents of the present invention at the 9th and 2α positions of sialic acid were extremely effective in inhibiting neuraminidase activity.

Formulation Example 1
A granule containing the following ingredients is produced.
Component Compound represented by formula (I) 10 mg
Lactose 700mg
Corn starch 274mg
HPC-L 16mg
1000mg total
The compound of formula (I) and lactose are passed through a 60 mesh sieve. Pass corn starch through a 120 mesh sieve. These are mixed in a V-type mixer. An HPC-L (low viscosity hydroxypropylcellulose) aqueous solution is added to the mixed powder, followed by kneading, granulation (extruded granulation, pore diameter: 0.5 to 1 mm), and drying step. The obtained dried granules are combed with a vibration sieve (12/60 mesh) to obtain granules.

Formulation Example 2
A capsule filling granule containing the following ingredients is produced.
Ingredient Compound represented by formula (I) 15 mg
Lactose 90mg
Corn starch 42mg
HPC-L 3mg
150mg total
The compound of formula (I), lactose, is passed through a 60 mesh sieve. Pass corn starch through a 120 mesh sieve. These are mixed, the HPC-L solution is added to the mixed powder, kneaded, granulated and dried. After the resulting dried granules are sized, 150 mg thereof is filled into a No. 4 hard gelatin capsule.

Formulation Example 3
A tablet containing the following ingredients is produced.
Component Compound represented by formula (I) 10 mg
Lactose 90mg
Microcrystalline cellulose 30mg
CMC-Na 15mg
Magnesium stearate 5mg
150mg total
The compound of formula (I), lactose, microcrystalline cellulose, CMC-Na (carboxymethylcellulose sodium salt) are passed through a 60 mesh sieve and mixed. The mixed powder is mixed with magnesium stearate to obtain a mixed powder for tableting. The mixed powder is directly hit to obtain a 150 mg tablet.

Formulation Example 4
The following components were heated and mixed and then sterilized to give an injection.
Component Compound represented by formula (I) 3 mg
Nonionic surfactant 15mg
Purified water for injection 1ml

5-Acetamide-9-azido-3,5,9-trideoxy-2-S- (4-nitrophenyl) -D-glyceo-α-D-galacto-non-2-uropyranosonic acid synthesized in Synthesis Example 1 ( It is a figure which shows the manufacturing method of compound 1).

Claims (9)

Formula (I)

(Wherein R ¹ and R ² each independently represents a substituent having an aromatic ring), or a pharmaceutically acceptable salt thereof, or a solvate thereof. Formula (II)

(In the formula, R ³ represents an aliphatic group which may have a substituent or an aromatic group which may have a substituent, and X is a single bond, —S—, — (CH ₂ ). _n — (wherein _n represents a natural number of 1 to 5), —CH═CH— or —C≡C—, and R ⁴ represents an aromatic group or aromatic group which may have a substituent. Or a pharmaceutically acceptable salt thereof, or a solvate thereof. The compound according to claim 1, which represents an aliphatic group having a ring. R ³ has an aryl group which may have a substituent, an aliphatic ring group which may have a substituent, a heteroaromatic group thioalkyl group which may have a substituent, or a substituent. Or a pharmaceutically acceptable salt thereof or a solvate thereof. The compound according to claim 2, which is an arylcarbamoylalkyl group which may be substituted. Formula (I)

(Wherein R ¹ and R ² each independently represents a substituent having an aromatic ring) or a pharmaceutical comprising a pharmaceutically acceptable salt or solvate thereof as an active ingredient Composition. Formula (II)

(In the formula, R ³ represents an aliphatic group which may have a substituent or an aromatic group which may have a substituent, and X is a single bond, —S—, — (CH ₂ ). _n — (wherein _n represents a natural number of 1 to 5), —CH═CH— or —C≡C—, and R ⁴ represents an aromatic group or aromatic group which may have a substituent. The pharmaceutical composition according to claim 4, which represents an aliphatic group having a ring. R ³ has an aryl group which may have a substituent, an aliphatic ring group which may have a substituent, a heteroaromatic group thioalkyl group which may have a substituent, or a substituent. The pharmaceutical composition according to claim 5, which is an optionally substituted arylcarbamoylalkyl group. Formula (I)

(Wherein R ¹ and R ² each independently represents a substituent having an aromatic ring), a sialidase inhibitor comprising the compound represented by the formula or a pharmaceutically acceptable salt thereof or a solvate thereof. Formula (II)

(In the formula, R ³ represents an aliphatic group which may have a substituent or an aromatic group which may have a substituent, and X is a single bond, —S—, — (CH ₂ ). _n — (wherein _n represents a natural number of 1 to 5), —CH═CH— or —C≡C—, and R ⁴ represents an aromatic group or aromatic group which may have a substituent. The sialidase inhibitor according to claim 7, which represents an aliphatic group having a ring. R ³ has an aryl group which may have a substituent, an aliphatic ring group which may have a substituent, a heteroaromatic group thioalkyl group which may have a substituent, or a substituent. The sialidase inhibitor according to claim 8, which is an optionally substituted arylcarbamoylalkyl group.

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