CN1261894A - Polyol-amino-acid compounds having anti-helicobactor pylori activity - Google Patents

Abstract

Formula (I) compound, wherein R ¹ represents a substitutable amino group; R ² represents a carboxyl group that can be esterified or amidated; R ³ , R ⁴ , R ⁵ , and R ⁶ represent a hydroxy group that can be protected; Q represents a substitutable aryl; or a salt thereof. The compound (I) has anti-helicobacter pylori activity and can be used for preventing or treating various diseases related to helicobacter pylori, such as duodenal ulcer, gastric ulcer, chronic gastric ulcer and gastric cancer.

Description

Polyhydroxy-amino acid compounds with anti-helicobacter pylori activity

technical field

The present invention relates to a polyhydroxy compound, its preparation method and application. More specifically, the present invention relates to a biologically active compound useful as a drug such as a prophylactic and therapeutic drug for gastric ulcer and duodenal ulcer, and an anti-Helicobacter pylori (hereinafter abbreviated as H. pylori or HP) agent containing the compound .

Background technique

A member of the noxious flora in the gastrointestinal tract, H. pylori is a Gram-negative microaerophilic bacterium belonging to the Helicobacter pylori genus; it has been suggested that it may be a major factor in gastritis, duodenal ulcer, and gastric ulcer recurrence.

For the treatment of various diseases associated with Helicobacter pylori infection, chemotherapy is used today, such as double drug combination therapy using bismuth and antibiotics or using bismuth, metronidazole (US Pat. Patent 2,712,517) or amoxicillin (US Patent 3,192,198) triple drug combination therapy. A triple therapy consisting of a gastric proton pump inhibitor, amoxicillin, and clarithromycin has also been found to be effective (Gut, 1995, 37 (Appendix 1): A365) (Gastroenterology, 1996, 110: A171) . Medications such as bismuth, antibiotics, and metronidazole are all given by mouth.

以及Carbohyd.Res.，28(2)，263-280(1973)指出能有效抑制革兰氏阴性菌。For polyols, PCT International Patent Application Publication No. WO93/06838 and ActaChemical Scandinavica B36, 515-518 (1982) disclose the compound of the following formula as a synthetic intermediate and

And Carbohyd.Res., 28(2), 263-280(1973) pointed out Can effectively inhibit Gram-negative bacteria.

In order to improve the efficient expression of the active ingredient and reduce the risk of side effects, attempts have been made to formulate amoxicillin, e.g. in gastric mucoadhesive compositions, to prolong its gastric retention time and allow amoxicillin to be released at a controlled rate, thereby improving activity Availability of ingredients (WO94/00112). It has been shown that H. pylori clearance can be improved by allowing anti-H. pylori substances to remain in the stomach for a prolonged period of time to ensure prolonged exposure of the bacteria to the active substance. [Scand. J. Gastroenterol., 29, 16-42 (1994)].

However, in order to maintain a sufficient growth inhibitory concentration in the habitat of Helicobacter pylori, the bismuth, antibiotic or metronidazole must be administered daily in large doses, and this treatment has various problems such as vomiting and diarrhea etc. Side effects occur. In this context, the object of the present invention is to provide a new drug which has high antibacterial activity, especially against Helicobacter pylori and other bacteria of the Helicobacter genus, and produces clinically beneficial prophylactic and therapeutic responses while reducing negative The emergence of the reaction.

Summary of invention

In this context, the object of the present invention is to provide a new drug which has high antibacterial activity, especially against Helicobacter pylori and other bacteria of the Helicobacter genus, and produces clinically beneficial prophylactic and therapeutic responses while reducing negative The emergence of the reaction.

The object of the present invention is to provide a pharmaceutical composition which has enhanced mucoadhesive activity compared to other gastric mucoadhesive preparations, whereby the efficiency of the active ingredient is greatly improved, in particular, to provide an anti-pyloric screw A bacterial composition and a pharmaceutical preparation for preventing, treating or preventing duodenal ulcer recurrence; it is very satisfactory and favorable in terms of anti-helicobacter pylori effect, low risk of side effects, sustained effect and safety.

[其中R¹代表可取代的氨基；R²代表可酯化或酰胺化的羧基；R³，R⁴，R⁵，和R⁶分别代表可保护的羟基；Q代表可取代的芳基]，其结构的特殊之处在于下式定义的基团

[Q和R²定义如上]直接键联到碳原子上，而且发现由于独特的化学结构，上述化合物对有害于胃肠道的菌类显示出显著的抑制活性，特别是高度的抗幽门螺杆菌活性，同时具有临床有利的药理活性，例如低的副作用的危险。本发明是基于上述发现完成的。The present inventor is through in-depth study, the result synthesizes the new polyhydric alcohol (polyhydroxyl compound) of following formula:

[Where R ¹ represents a substitutable amino group; R ² represents a carboxyl group that can be esterified or amidated; R ³ , R ⁴ , R ⁵ , and R ⁶ represent a protective hydroxyl group; Q represents a substitutable aryl group], The special feature of its structure is the group defined by the following formula

[Q and ^R2 as defined above] are directly bonded to the carbon atom, and it was found that due to the unique chemical structure, the above compound shows significant inhibitory activity against bacteria harmful to the gastrointestinal tract, especially highly anti-H. pylori activity, while having clinically favorable pharmacological activities, such as a low risk of side effects. The present invention has been accomplished based on the above findings.

其中R¹代表可取代的氨基；R²代表可酯化或酰胺化的羧基；R³，R⁴，R⁵，和R⁶分别代表可保护的羟基；Q代表可取代的芳基；或其盐。In view of the state of the prior art described above, the present inventors have found that the effectiveness of the active ingredient (e.g. anti-Helicobacter pylori action) can be improved by adding a enhanced by agents that swell viscous agents such as alcaligenes polysaccharide (curdlan) and/or low-substituted hydroxypropyl cellulose; the composition was also found to have a reliable safety profile and enhanced adhesion to mucous membranes . Therefore, the present invention relates to: 1. the compound of formula (I)

Wherein R ¹ represents a substitutable amino group; R ² represents a carboxyl group that can be esterified or amidated; R ³ , R ⁴ , R ⁵ , and R ⁶ represent a protective hydroxyl group; Q represents a substitutable aryl group; or Salt.

2. A compound according to paragraph 1 above, wherein R ¹ is amido or amino substituted by a substitutable hydrocarbyl.

3. The compound according to item 2 above, wherein the amido group is an amino group substituted by an amino acid residue.

4. The compound according to item 3 above, wherein the amino acid residue is an α-amino acid residue.

5. According to the compound of paragraph 1 above, wherein R ¹ is an amino group or a group of the following formula: Wherein R ⁷ is an amino group substituted by an α-L-amino acid residue that can be substituted by an α-L-amino acid residue, R ⁸ is a substitutable hydrocarbon group; R ² represents a carboxyl group; R ³ , R ⁴ , R ⁵ , and R ⁶ represents hydroxyl respectively; Q represents phenyl.

6. The compound according to item 5 above, which is represented by the following formula (V): Where R ⁷ and R ⁸ have the same definition as in paragraph 5 above.

7. According to the compound of paragraph 5, wherein R ⁸ is C _1-10 alkyl, C _6-14 aryl-C _1-6 alkyl, C _2-10 alkenyl or C _2-10 alkynyl, these Groups can be substituted.

8. The compound according to paragraph 7 above, wherein R ⁸ is C _1-6 alkyl or C _2-6 alkenyl.

9. The compound according to paragraph 7 above, wherein R ⁷ is amino which may be substituted by valyl, valylvalyl, valylisoleucyl or valylleucyl.

10. The compound according to paragraph 8 above, wherein R ⁸ is isobutyl or allyl.

11. The compound according to paragraph 1 above, wherein R ¹ is amino.

12. According to the compound of paragraph 1 above, it is (S)-3-[(2S, 3R, 4R, 5S)-2,3,4,6-tetrahydroxy-5-(L-valyl-L- Valyl-L-leucyl)aminocaproyl]amino-3-phenylpropanoic acid.

13. According to the compound of paragraph 1 above, it is (S)-3-[(2S, 3R, 4R, 5S)-2,3,4,6-tetrahydroxy-5-(L-valyl-L- Isoleucyl-L-leucyl)aminocaproyl]amino-3-phenylpropanoic acid.

14. According to the compound of paragraph 1 above, it is (S)-3-[(2S, 3R, 4R, 5S)-2,3,4,6-tetrahydroxy-5-(L-valyl-L- Leucyl-L-leucyl)aminocaproyl]amino-3-phenylpropanoic acid.

15. According to the compound of paragraph 1 above, it is (S)-3-[(2S, 3R, 4R, 5S)-2,3,4,6-tetrahydroxy-5-(L-valyl-L- Leucyl)aminocaproyl]amino-3-phenylpropanoic acid.

16. According to the compound of paragraph 1 above, it is (S)-3-[(2S, 3R, 4R, 5S)-2,3,4,6-tetrahydroxy-5-(L-leucyl)aminohexyl Acyl]amino-3-phenylpropanoic acid.

17. According to the compound of paragraph 1 above, it is (S)-3-[(2S, 3R, 4R, 5S)-5-((S)-2-amino-4-pentenoyl)amino-2,3 , 4,6-Tetrahydroxyhexanoyl]amino-3-phenylpropanoic acid.

18. According to the compound of paragraph 1 above, it is (S)-3-[(2S, 3R, 4R, 5S)-5-((S)-2-aminobutyryl)amino-2,3,4,6 -tetrahydroxyhexanoyl]amino-3-phenylpropanoic acid.

19. According to the compound of paragraph 1 above, it is (S)-3-[(2S, 3R, 4R, 5S)-2,3,4,6-tetrahydroxy-5-(L-isoleucyl-L -leucyl)aminocaproyl]amino-3-phenylpropanoic acid.

20. According to the compound of paragraph 1 above, it is (S)-3-[(2S, 3R, 4R, 5S)-2,3,4,6-tetrahydroxy-5-(L-methionyl-L -leucyl)aminocaproyl]amino-3-phenylpropanoic acid.

21. According to the compound of paragraph 1 above, it is (S)-3-[(2S, 3R, 4R, 5S)-2,3,4,6-tetrahydroxy-5-((S)-2-(L -norvalyl)amino-4-pentenoyl)aminocaproyl]amino-3-phenylpropanoic acid.

22. A pharmaceutical composition comprising a compound according to item 1 above.

23. The composition according to item 22 above, which is an anti-Helicobacter pylori agent.

24. The anti-Helicobacter pylori agent according to 23 above, which is a preventive and therapeutic agent for diseases related to Helicobacter pylori infection.

25. The anti-Helicobacter pylori agent according to item 24 above, wherein the disease associated with Helicobacter pylori infection is gastric ulcer or duodenal ulcer, gastritis, gastric cancer or gastric MALT lymphoma.

26. An anti-Helicobacter pylori agent comprising the compound according to item 1 above in combination with at least one other antibacterial agent or/and antiulcer agent.

27. The composition according to item 22 above, which is a gastric mucosa-adhesive pharmaceutical composition.

28. The composition according to item 27 above, which comprises (a) the compound according to item 1 above, (b) lipid and/or polyglycerol fatty acid ester and (c) a viscous agent capable of forming viscosity with water.

29. The composition according to item 28 above, wherein (c) the viscous agent is an acrylic polymer or a salt thereof.

30. The composition according to item 28 above, which comprises (d) a substance capable of swelling the viscous agent.

31. The composition according to item 30 above, wherein the substance capable of swelling the viscous agent is Alcaligenes polysaccharide and/or low-substituted hydroxypropylcellulose.

其中R¹是可取代的氨基；R³，R⁴，R⁵，和R⁶分别代表可保护的羟基，或其盐，或其活性衍生物；与式(III)化合物

其中R²代表可酯化或酰胺化的羧基；Q代表可取代的芳基，或其盐进行反应。32. A method for the preparation of a compound according to the preceding paragraph 1, which comprises making the carboxylic acid of formula (II)

Wherein R ¹ is a substitutable amino group; R ³ , R ⁴ , R ⁵ , and R ⁶ represent respectively a protectable hydroxyl group, or a salt thereof, or an active derivative thereof; and the compound of formula (III)

Wherein R ² represents a carboxyl group that can be esterified or amidated; Q represents a substitutable aryl group, or a salt thereof for reaction.

其中R²代表可酯化或酰胺化的羧基；R³，R⁴，R⁵和R⁶分别代表可保护的33. A method for the preparation of a compound according to paragraph 1, which comprises making the compound of formula (IV)

Where R ² represents a carboxyl group that can be esterified or amidated; R ³ , R ⁴ , R ⁵ and R ⁶ represent a carboxyl group that can be protected

Hydroxy, Q represents a substitutable aryl group, or a salt thereof, or an active derivative thereof; and a compound of the following formula:

R ⁹ -X wherein R ⁹ represents an acyl group, or a substituted hydrocarbon group; X represents a leaving group, or a salt thereof, or a reactive derivative thereof.

34. A method for preparing the compound according to the above paragraph 5, which comprises growing a microbial strain of the genus Bacillus (Bacillus) capable of producing the compound according to the above paragraph 5 in a medium, allowing the bacterial strain to grow and accumulate in a fermentation broth The compound, and collecting the compound.

35. The method according to item 34 above, wherein the microbial strain is Bacillus insolitus HC-70 or Bacillus insolitus HC-72.

36. Bacillus HC-70 or Bacillus anomaly HC-72 capable of producing the compound according to item 5 above.

其中R¹代表可取代的氨基；R²代表可酯化或酰胺化的羧基；R³，R⁴，R⁵，和R⁶分别代表可保护的羟基；Q代表可取代的芳基；或其盐。37. A method for preventing or treating diseases associated with Helicobacter pylori infection in mammals, comprising administering an effective amount of a compound of formula (I) to a mammal in need

Wherein R ¹ represents a substitutable amino group; R ² represents a carboxyl group that can be esterified or amidated; R ³ , R ⁴ , R ⁵ , and R ⁶ represent a protective hydroxyl group; Q represents a substitutable aryl group; or Salt.

其中R¹代表可取代的氨基；R²代表可酯化或酰胺化的羧基；R³，R⁴，R⁵，和R⁶分别代表可保护的羟基；Q代表可取代的芳基；或其盐。38. The compound of formula (I) is used to prepare the purposes of anti-helicobacter pylori agent

Wherein R ¹ represents a substitutable amino group; R ² represents a carboxyl group that can be esterified or amidated; R ³ , R ⁴ , R ⁵ , and R ⁶ represent a protective hydroxyl group; Q represents a substitutable aryl group; or Salt.

A brief description of the drawings

Fig. 1 is the ¹³ C-NMR spectrum of HC-70I obtained in Example 2.

FIG. 2 is a ¹³ C-NMR spectrum of HC-70II obtained in Example 1. FIG.

FIG. 3 is a ¹³ C-NMR spectrum of HC-70III obtained in Example 1. FIG.

The "substitutable amino group" mentioned for ^R1 includes amino group, amido group, and amino group substituted by a substitutable hydrocarbon group.

The "acyl" in the "acylamino" mentioned by ^R1 not only includes any amino acid residue or a sequence of two or more "amino acid residues" (hereinafter referred to as the "amino acid of R ^a , R ^b or R ^c residue"), but also includes, among other acyl groups, optionally substituted alkanoyl, substituted aroyl, substituted heterocyclic carbonyl, substituted carbamoyl, substituted thiocarbamoyl, substituted Alkylsulfonyl, optionally arylsulfonyl, optionally sulfamoyl, optionally alkoxycarbonyl, and optionally substituted aryloxycarbonyl.

Among them, a sequence of two or more "amino acid residues" is particularly preferred.

The "alkanoyl" of the "substitutable alkanoyl" includes but is not limited to C _1-20 alkanoyl (for example, formyl, acetyl, propionyl, isopropionyl, butyryl, pentanoyl, hexanoyl, heptanoyl, acyl, octanoyl, nonanoyl, lauroyl, undecanoyl, myristoyl, palmitoyl, stearoyl, etc.), and particularly preferably C _1-6 alkanoyl (for example, formyl, acetyl, propionyl , isopropionyl, butyryl, valeryl, and hexanoyl).

The "aroyl" of the "substitutable aroyl" includes, but is not limited to, C _7-16 aroyl (eg, benzoyl, 1-naphthoyl, 2-naphthoyl, etc.).

The "heterocyclic carbonyl" of the "substitutable heterocyclic carbonyl" includes 5- or 6-membered ones containing 1 to 4 heteroatoms (for example, nitrogen, oxygen and sulfur), respectively, in addition to carbon as ring atoms. Heterocyclic carbonyl or fused heterocyclic carbonyl (for example, 3-pyrrolecarbonyl, 1-imidazolecarbonyl, 1-pyrazolecarbonyl, 3-isothiazolecarbonyl, 3-isoxazolecarbonyl, pyrazinecarbonyl, 2-pyrimidinecarbonyl, 3-pyrazinecarbonyl, 2-indenenitrocarbonyl, 2-isoindolecarbonyl, 1-indolecarbonyl, 2-furoyl, 2-thienyl, nicotinoyl, isonicotinoyl, morpholinocarbonyl, piperidinocarbonyl, piperazinecarbonyl, etc.).

The "alkylsulfonyl" of the "substitutable alkylsulfonyl" includes but is not limited to C _1-20 alkylsulfonyl (for example, methylsulfonyl, ethylsulfonyl, propylsulfonyl, isopropyl sulfonyl, etc.).

"Substitutable carbamoyl" includes not only carbamoyl, but also mono-substituted carbamoyl and di-substituted carbamoyl, wherein the substituents can be selected from, for example, C _1-6 alkyl (for example, methyl base, ethyl, propyl, isopropyl, butyl, isobutyl, etc.), C _6-14 aryl (for example, phenyl, 1-naphthyl, 2-naphthyl, etc.), C _7- Aralkyl (for example, benzyl, etc.), C _1-6 alkanoyl (for example, acetyl, propionyl, isopropionyl, butyryl, _etc. ), C _7-16 aroyl (for example, benzoyl , 1-naphthoyl, 2-naphthoyl, etc.), 5- or 6-membered heterocyclic carbonyl (for example, 3-pyrrolecarbonyl, 2-imidazolecarbonyl, 1-pyrazolecarbonyl, 3-isothiazolylcarbonyl, 3- Isoxazole carbonyl, pyrazine carbonyl, 2-pyrimidine carbonyl, 3-pyrazine carbonyl, 2-indene nitrogen carbonyl, 2-isoindole carbonyl, 1-indole carbonyl, 2-furoyl, 2-thienyl acyl, nicotinoyl, isonicotinoyl, morpholinocarbonyl, piperidinocarbonyl, piperazinecarbonyl, etc.).

"Substitutable thiocarbamoyl" includes not only thiocarbamoyl, but also mono-substituted thiocarbamoyl and di-substituted thiocarbamoyl, wherein the substituents may be selected from, for example, C _{1- 6} alkyl (for example, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, etc.), C _6-14 aryl (for example, phenyl, 1-naphthyl, 2-naphthyl etc.), C _7-16 aralkyl (for example, phenyl-C _1-5 alkyl, such as benzyl, etc.), C _1-6 alkanoyl (for example, acetyl, propionyl, isopropionyl, butyryl, etc.), C _7-16 aroyl (for example, benzoyl, 1-naphthoyl, 2-naphthoyl, etc.), 5- or 6-membered heterocyclic carbonyl (for example, except for carbon as ring atoms In addition, 5- or 6-membered heterocyclic carbonyl or fused heterocyclic carbonyl containing 1-4 heteroatoms (such as nitrogen, oxygen and sulfur) respectively), such as 3-pyrrolecarbonyl, 2-imidazolecarbonyl, 1- Pyrazolecarbonyl, 3-isothiazolecarbonyl, 3-isoxazolecarbonyl, pyrazinecarbonyl, 2-pyrimidinecarbonyl, 3-pyrazinecarbonyl, 2-indenenitrocarbonyl, 2-isoindolecarbonyl, 1-indole carbonyl, 2-furoyl, 2-thienoyl, nicotinoyl, isonicotinoyl, morpholinocarbonyl, piperidinocarbonyl, piperazinecarbonyl, etc.).

The "arylsulfonyl" of the "substitutable arylsulfonyl" includes but is not limited to C _6-14 arylsulfonyl (for example, benzenesulfonyl, 1-naphthylsulfonyl, 2-naphthylsulfonyl etc).

"Substitutable sulfamoyl" includes not only sulfamoyl, but also mono-substituted sulfamoyl and di-substituted sulfamoyl, wherein the substituents can be selected from, for example, C _1-6 alkyl (e.g., methyl , ethyl, propyl, isopropyl, butyl, isobutyl, etc.), C _6-14 aryl (eg, phenyl, 1-naphthyl, 2-naphthyl, etc.) and C _7-16 Aralkyl (eg, phenyl-C _1-5 alkyl, eg benzyl, etc.).

The "alkoxycarbonyl" of the "substitutable alkoxycarbonyl" includes but is not limited to C _1-20 alkoxycarbonyl (for example, methoxycarbonyl, ethoxycarbonyl, propoxycarbonyl, isopropoxycarbonyl , butoxycarbonyl, isobutoxycarbonyl, etc.).

The "aryloxycarbonyl" of the "substitutable aryloxycarbonyl" includes but is not limited to C _6-14 aryloxycarbonyl (for example, phenoxycarbonyl, 1-naphthyloxycarbonyl, 2-naphthyloxy carbonyl, etc.).

The "hydrocarbon group" of the "substitutable hydrocarbon group" may be, for example, an aliphatic hydrocarbon group or a cyclic hydrocarbon group. The above-mentioned "aliphatic hydrocarbon group" includes, but is not limited to, a C _1-20 aliphatic hydrocarbon group (for example, an alkyl group, an alkenyl group and an alkynyl group). "Cycloalkyl" includes C _3-20 cycloalkyl (eg, cycloalkyl, cycloalkenyl, aryl, etc.).

The above-mentioned "alkyl" is a C _1-10 alkyl group, such as methyl, ethyl, propyl, 2-propyl, 1-ethylpropyl, butyl, 1-methylpropyl, 2-methylpropyl base, 1,1-dimethylethyl, 1,1-dimethylbutyl, 2,2-dimethylbutyl, pentyl, 3-methylbutyl, 2,2-dimethylpropane base, hexyl, etc.

"Alkenyl" is C _2-10 alkenyl, such as vinyl, 2-propenyl, 1-methylvinyl, butenyl, 2-methyl-1-propenyl, 2-methyl-2 -propenyl, 1-methyl-2-propenyl, 2-pentenyl, 3-pentenyl, 4-pentenyl, 2-hexenyl, 3-hexenyl, 5-hexenyl, etc. wait.

"Alkynyl" is C _2-10 alkynyl, such as ethynyl, 2-propynyl, 2-butyn-1-yl, 3-butyn-2-yl, 1-pentyn-3-yl, 3 -pentyn-1-yl, 4-pentyn-2-yl, 3-hexyn-1-yl and the like.

"Cycloalkyl" is C _3-10 cycloalkyl, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and the like.

"Cycloalkenyl" is C _3-10 cycloalkenyl, such as cyclobutenyl, cyclopentenyl, cyclohexenyl, cyclohexadienyl and the like.

"Aryl" is C _6-14 aryl, such as phenyl, 1-naphthyl, 2-naphthyl and the like.

For the "hydrocarbyl" of the "substitutable hydrocarbyl" of ^R and the 'acyl' of the "acylamino", for the "alkanoyl, aroyl, heterocyclic carbonyl, alkylsulfonyl, arylsulfonyl, The optionally substituted substituents on the "alkoxycarbonyl or aryloxycarbonyl" are not particularly limited unless inconsistent with the purpose of the present invention, so they include amino, mono- or di-C _1-6 alkylamino (such as methylamino, ethylamino, propylamino, isopropylamino, dimethylamino, diethylamino, etc.), one- or two-C _6-10 arylamino (for example, aniline, diphenylamino, etc.), one - or di-C _7-11 aralkylamino (for example, phenyl-C _1-5 alkylamino such as benzylamino, di(phenyl C _1-5 alkyl) amino such as dibenzylamino, etc.) , azido, nitro, halogen (for example, fluorine, chlorine, bromine, iodine), hydroxyl, C _1-6 alkoxy (for example methoxy, ethoxy, propoxy, isopropoxy, butyl Oxy, etc.), C _6-10 aryloxy (phenoxy, 1-naphthyloxy, 2-naphthyloxy, etc.), C _7-11 aralkoxy (for example, phenyl-C _{1- 5} alkoxy such as benzyloxy, etc.), formyloxy, C _1-6 alkylcarbonyloxy (such as acetyloxy, propionyloxy, etc.), C _6-10 arylcarbonyloxy ( For example, benzoyloxy, etc.), C _7-11 aralkylcarbonyloxy (for example, phenyl-C _1-5 alkylcarbonyloxy such as benzylcarbonyloxy, etc.), sulfonyloxy , C _1-6 alkylsulfonyloxy (for example, methylsulfonyloxy, etc.), mercapto, C _1-6 alkylthio (for example, methylthio, ethylthio, propylthio, isopropyl Thio, etc.), C _6-10 arylthio (for example, phenylthio, 1-naphthylthio, 2-naphthylthio, etc.), C _7-11 aralkylthio (for example, phenyl- C _1-5 alkylthio such as benzylthio etc.), phosphonooxy, cyano, carbamoyl, mono- or di-C _1-6 alkylcarbamoyl (for example, methylcarbamoyl , ethylcarbamoyl, dimethylcarbamoyl, diethylcarbamoyl, etc.), one- or di-C _6-10 arylcarbamoyl (for example, phenylcarbamoyl, diphenyl carbamoyl, etc.), mono- or di-C _7-11 aralkylcarbamoyl (for example (phenyl-C _1-5 alkyl) carbamoyl such as benzylcarbamoyl, di(phenyl- C _1-5 alkyl)carbamoyl such as dibenzylcarbamoyl, etc.), carboxyl, C _1-6 alkoxycarbonyl (for example, methoxycarbonyl, ethoxycarbonyl, etc.), C _6-10 aryloxy Carbonyl (for example, phenoxycarbonyl, 1-naphthyloxycarbonyl, 2-naphthyloxycarbonyl, etc.), C _7-11 aralkyloxycarbonyl (for example, phenyl-C _1-5 alkoxycarbonyl such as benzyloxycarbonyl, etc. etc.), formyl, C _1-6 alkylcarbonyl (such as acetyl, propionyl, isopropionyl, butyryl, pentanoyl, hexanoyl, etc.), C _6-10 arylcarbonyl (such as benzoyl , 1-naphthoyl, 2-naphthoyl, etc.), C _7-11 aralkylcarbonyl (for example, phenyl-C _1-5 alkylcarbonyl such as benzylcarbonyl, etc.), sulfo, C _1-6 Alkylsulfinyl (for example, methylsulfinyl, ethylsulfinyl, etc.), C _6-10 arylsulfinyl (for example, phenylsulfinyl, 1-naphthylsulfinyl, 2- naphthylsulfinyl, etc.), C _1-6 alkylsulfonyl (for example, methylsulfonyl, ethylsulfonyl, etc.), C _6-10 arylsulfonyl (for example, benzenesulfonyl, 1-naphthalene sulfonyl, 2-naphthylsulfonyl, etc.), C _1-6 alkyl (for example, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, etc.), C _2-6 alkenyl (e.g. vinyl, allyl, 2-butenyl, etc.), C _2-6 alkynyl (e.g., ethynyl, propynyl, etc.), C _3-6 cycloalkane (for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, etc.), C _3-6 cycloalkenyl (for example, cyclobutenyl, cyclopentenyl, cyclohexenyl, cyclohexanedi alkenyl, etc.), C _6-10 aryl (for example, phenyl, 1-naphthyl, 2-naphthyl, etc.), one-to three-ring heterocyclic group (for example, consisting of 1-3 containing A heterocyclic group consisting of at least one 5- or 6-membered ring of 1 to 4 heteroatoms selected from nitrogen, oxygen and sulfur: pyridyl, pyrazinyl, pyrimidinyl, quinolinyl, isoquinoline base, indolyl, isoindolyl, indazolyl, pyridazinyl, imidazolyl, pyrazolyl, pyrrolyl, furyl, benzofuryl, thienyl, benzothienyl, benzimidazolyl, Quinazolinyl, pyrrolidinyl, pyrrolinyl, imidazolidinyl, imidazolinyl, pyrazolidinyl, pyrazolinyl, piperidinyl, piperazinyl, indenyl, isoindenyl, morpholinyl, etc.), and mono- to tri-ring heterocyclic thio groups (for example, groups formed by bonding a thio group to the above-mentioned heterocyclic groups, for example, 4-piperidinylthio, 2-pyrimidinylthio group, 1,3,4-thiadiazol-2-ylthio group, 5-tetrazolylthio group, 2-benzothiazolylthio group, 8-quinolinethio group, etc.). These substituents may be present on the "hydrocarbyl" and the "alkanoyl, aroyl, heterocyclic carbonyl, alkylsulfonyl, arylsulfonyl, alkoxycarbonyl, and aryloxycarbonyl" within the chemically permitted range. , and in each case the number of substituents may be 1-5, preferably 1-3. It should be understood that when the number of substituents is 2 or more, the substituents may be similar or not. Where chemically permitted, these substituents may also be substituted with 1-3 substituents selected from amino, mono- or di-C _1-6 alkylamino, nitro, halogen, hydroxy, C _{1 -6} alkoxy, C _1-6 alkylcarbonyloxy, sulfonyloxy, C _1-6 alkylsulfonyloxy, mercapto, C _1-6 alkylthio, phosphonyloxy, cyano, Carbamoyl, mono- or di-C _1-6 alkylcarbamoyl, carboxy, C _1-6 alkoxycarbonyl, formyl, C _1-6 alkylcarbonyl, sulfo, and C _1-6 alkyl Sulfinyl.

R ¹ further includes groups of the following formula: R ^a -R ^b -R ^c -NH-[R ^a represents hydrogen or an amino acid residue that can be replaced; R ^b and R ^c can be the same or different, and represent a bond respectively, which can be substituted Amino acid residues, or YR ^d -(R ^d represents the group obtained by removing imino groups from substitutable amino acid residues; Y represents -O-, -S-, or -NR ^e -(R ^e represents hydrogen or lower alkyl))].

Provided that R ^a , R ^b and/or R ^c are amino acid residues, they are preferably linked together by an amide bond.

The above-mentioned 'amino acid' in the "amino acid residue" of R ^a , R ^b and R ^c and the "group obtained by removing the imino group ^of the amino acid residue" of R ^d and the "amino acid residue substituted by the amino acid residue" of R It generally means a group obtained by replacing at least one hydrogen atom in the core structure of a carboxylic acid with an amino group, including α-, β-, γ- and δ-amino acids with a core structure of 2-20 carbon atoms. Among these amino acids, α-amino acids (especially α-L-amino acids) are preferred, such as alanine, arginine, asparagine, aspartic acid, cysteine, glutamic acid, glutamine, Glycine, Histidine, Leucine, Isoleucine, Lysine, Methionine, Phenylalanine, Proline, Serine, Threonine, Tryptophan, Tyrosine, Valine acid, etc., and other amino acids such as norvaline, norleucine, 2-aminoadipic acid, 2-aminobutyric acid, 2-aminoisobutyric acid, 2-amino-4-pentenoic acid, 1 -Aminocyclopropanecarboxylic acid, 1-aminocyclopentanecarboxylic acid, 1-aminocyclohexanecarboxylic acid, thyronine, ornithine, hydroxyproline, hydroxylysine, (2-naphthyl) Alanine, Azaglycine and more.

等等。优选脯氨酸，羟基脯氨酸和六氢哌啶羧酸。The aforementioned amino acids include cyclic imino acids. "Cyclic imino acid" means a compound obtained by substituting an imino group for at least one methylene group of a cycloalkane carboxylic acid or a cycloalkene carboxylic acid, thus they include proline, hydroxyproline, 3,4-dehydroproline amino acid, hexahydropiperidine carboxylic acid Aziridinecarboxylic acid, and 2-azetidinecarboxylic acid

etc. Preference is given to proline, hydroxyproline and hexahydropiperidinecarboxylic acid.

The term amino acid used in the specification may be any amino acid residue commonly used in peptide chemistry, for example may be any of the aforementioned amino acid residues.

The "group obtained by removing an imino group (-NH-) from an amino acid residue" of R ^d may be a group obtained by removing an imino group from the above-mentioned amino acid residue, and therefore includes a carboxylic acid having the following group as a core Structurally derived groups: straight or branched _C1-10 alkyl (e.g., methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl , isopentyl, neopentyl, tert-pentyl, 1-ethylpropyl, hexyl, isohexyl, heptyl, octyl, decyl, etc.), C _2-10 alkenyl (for example, vinyl, Allyl, isopropenyl, 1-propenyl, 2-methyl-1-propenyl, 1-, 2- or 3-butenyl, 2-ethyl-1-butenyl, 3-methyl -2-butenyl, 1-, 2-, 3- or 4-pentenyl, 4-methyl-3-pentenyl, 1-, 2-, 3-, 4- or 5-hexenyl , and heptenyl, octenyl and decenyl with a double bond at any position respectively), C _7-20 aralkyl (for example, phenyl-C _1-5 alkyl such as benzyl, phenylethyl Base, 3-phenylpropyl, naphthyl-C _1-5 alkyl such as 1-naphthylmethyl, 2-naphthylmethyl, 9-fluorenylmethyl, etc.), C _3-7 cycloalkyl (such as , cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, etc.), C _3-7 cycloalkenyl (for example, 2-cyclopenten-1-yl, 3-cyclopenten-1 -yl, 2-cyclohexen-1-yl, 3-cyclohexen-1-yl, etc.), C _6-15 aryl (such as phenyl, naphthyl, anthracenyl, phenanthrenyl, acenaphthyl, fluorene etc.), and monocyclic or fused polycyclic C _3-20 heterocycloalkyl (for example, 4-imidazolylmethyl, 3-pyridylmethyl, 4-thiazolylmethyl, 3-indolylmethyl, 3-quinolinemethyl, etc.).

The above-mentioned amino acid residue and the above-mentioned group obtained by removing the imino group from the amino acid residue may have one or more substituents at appropriate positions, preferably 1-3 substituents, and the specific substituents that may exist include but not Limited to amino, acyl-substituted amino, guanidino, acylguanidino, acylamidino, amidino, acyl, carbamoyl, N-mono- or di-lower alkylcarbamoyl, carboxyl, lower alkoxycarbonyloxy radical, hydroxy, acylhydroxy, lower alkoxy, phenoxy, mercapto, acylmercapto, lower alkylthio, phenylthio, thio, cyano, azido, nitro, nitroso, and halogen.

Here, the "acyl" of the "acyl-substituted amino, acylguanidino, acylamidino, acylhydroxyl, or acylmercapto" includes the same acyl used for the "acyl" of the above-mentioned "acylamino" of ^R1 , especially C _1-20 alkanoyl (preferably C _1-6 alkanoyl).

The 'lower alkyl' of the "lower alkylcarbamoyl" includes but not limited to C _1-6 alkyl, such as methyl, ethyl, propyl, isopropyl, butyl, and isobutyl.

"Lower alkoxycarbonyloxy" includes but not limited to C _1-6 alkoxycarbonyloxy, such as methoxycarbonyloxy, ethoxycarbonyloxy, propoxycarbonyloxy, isopropoxycarbonyl oxy, butoxycarbonyloxy, isobutylcarbonyloxy, and the like.

"Lower alkoxy" includes but is not limited to C _1-6 alkoxy, such as methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy and the like.

"Lower alkylthio" includes, but is not limited to, C _1-6 alkylthio, such as methylthio, ethylthio, propylthio, isopropylthio and the like.

Lower alkyl groups for ^R include but are not limited to C _1-6 alkyl groups such as methyl, ethyl, propyl, isopropyl, butyl, pentyl, hexyl and the like.

The "α-L-amino acid" of R ⁷ in the "amino group which may be substituted by an α-L-amino acid residue" includes the same amino acids as previously described for R ^a , R ^b and R ^c An "α-L-amino acid" identical to an "α-L-amino acid" of residues.

The "substitutable hydrocarbon group" of R ⁸ includes the same examples of the substituents of the "substitutable amino group" of R ¹ above. Examples of ^R are C _1-10 alkyl, C _6-14 aryl-C _1-6 alkyl, C _2-10 alkenyl, C _2-10 alkynyl and the like, each of which can be was replaced. Preferred examples of R ⁸ are C _1-6 alkyl, C _2-6 alkenyl and cyclopropyl-C _1-3 alkyl, especially isobutyl and allyl.

Esterifiable carboxyl for R ² includes carboxyl, (lower (C _1-6 ) alkoxy) carbonyl (for example, methoxycarbonyl, ethoxycarbonyl, propoxycarbonyl, isopropoxycarbonyl, butoxy ylcarbonyl, isobutoxycarbonyl, tert-butoxycarbonyl, sec-butoxycarbonyl, pentyloxycarbonyl, isopentyloxycarbonyl, neopentyloxycarbonyl, tert-pentyloxycarbonyl, hexyloxycarbonyl, etc. ), (C _6-10 aryl)oxycarbonyl (for example, phenoxycarbonyl, 1-naphthyloxycarbonyl, etc.) and (C _7-10 aralkyl)oxycarbonyl (for example, (phenyl-C _{1 -4} alkoxy)carbonyl, such as benzyloxycarbonyl), diphenylmethoxycarbonyl, pivaloyloxymethoxycarbonyl, 1-(cyclohexyloxycarbonyloxy)ethoxycarbonyl. Among them, carboxyl, methoxycarbonyl and ethoxycarbonyl are preferred.

The amidatable carboxyl group for R ² includes carbamoyl and substituted carbamoyl. The substituents of the substituted carbamoyl group include, but are not limited to, unsubstituted or substituted lower (C _1-6 ) alkyl (for example, methyl, ethyl, propyl, isopropyl, butyl, isobutyl base, sec-butyl, tert-butyl, pentyl, isopentyl, neopentyl, hexyl, isohexyl, etc.), unsubstituted or substituted C _3-6 cycloalkyl (for example, cyclopropyl, cyclo butyl, cyclopentyl, cyclohexyl, etc.), unsubstituted or substituted C _6-10 aryl (for example, phenyl, 1-naphthyl, 2-naphthyl, etc.), unsubstituted or substituted C _7-12 aralkyl (for example, phenyl-C _1-4 alkyl such as benzyl and phenethyl, naphthyl-C _1-2 alkyl, etc.), and unsubstituted or substituted C _{6- 10} Arylsulfonyl (for example, benzenesulfonyl, 1-naphthalenesulfonyl, 2-naphthalenesulfonyl, etc.). There may be one or two similar or dissimilar groups selected from the above substituents. Such optionally substituted lower (C _1-6 )alkyl, optionally substituted C _3-6 cycloalkyl, optionally substituted C _6-10 aryl, optionally substituted C _7-12 aralkyl and optionally substituted Substituents for C _6-10 arylsulfonyl include halogen (for example, fluorine, chlorine, bromine, etc.), alkoxy groups that may be substituted by 1-3 halogens (for example, C _1-4 alkoxy such as methoxy , ethoxy, propoxy, etc.), alkyl which may be substituted by 1-3 halogens (for example, C _1-4 alkyl such as methyl, ethyl, propyl, etc.), amino, carboxyl and nitro and for these substituent groups there may be 1-5.

The substitutable amino group may be such a substituent that two substituents on the nitrogen atom form a cyclic amino group together with the nitrogen atom, and such cyclic amino groups include but are not limited to 1-azetidine, 1-pyrrolidine group, piperidino, morpholino, thiomorpholino, and 1-piperazinyl.

^R2 is preferably carboxyl.

The "aryl" of the "substitutable aryl" of Q includes, but is not limited to, monocyclic or fused polycyclic C _6-14 aromatic hydrocarbon groups. The above aromatic hydrocarbon groups include but are not limited to phenyl, 1- or 2-naphthyl, 1-, 2- or 9-anthracenyl, 1-, 2-, 3-, 4-, or 9-phenanthrenyl, and 1- , 2-, 4-, 5- or 6-azulene. This 'aryl' may have 1-5 suitable substituents at suitable positions, preferably 1-3 substituents, and such substituents include but are not limited to alkoxy (for example C _1-3 alkoxy such as methoxy, ethoxy and propoxy), halogen (for example, fluorine, chlorine, bromine, iodine), alkyl (for example, C _1-3 alkyl such as methyl, ethyl, and propyl), amino , nitro and cyano.

Q is preferably phenyl.

Protective groups for "protectable hydroxyl" of R ³ , R ⁴ , R ⁵ and R ⁶ include but are not limited to ether-forming protecting groups such as methoxymethyl, benzyloxymethyl, tert-butoxymethyl, 2-Methoxyethoxymethyl, 2-(trimethylsilyl)ethoxymethyl, methylthiomethyl, 2-tetrahydropyranyl, 4-methoxy-4-tetra Hydropyranyl, 2-tetrahydropyranyl, benzyl, p-methoxybenzyl, p-nitrobenzyl, o-nitrobenzyl, 2,6-dichlorobenzyl, trityl, etc. ; into a silyl ether protecting group, such as trimethylsilyl, triethylsilyl, triisopropylsilyl, isopropyldimethylsilyl, diethylisopropylsilyl base, tert-butyldimethylsilyl, tert-butyldiphenylsilyl, tribenzylsilyl, triphenylsilyl, methyldiphenylsilyl, etc.; and ester-forming Protecting groups such as formyl, acetyl, chloroacetyl, dichloroacetyl, trichloroacetyl, pivaloyl, benzoyl, benzyloxycarbonyl, 2-bromobenzyloxycarbonyl and the like. In addition to the above-mentioned protecting groups, protecting groups further include those groups having two hydroxyl groups among R ³ , R ⁴ , R ⁵ and R ⁶ , such as cyclic acetals such as methylene acetal, ethylene acetal, 4-Methoxyphenyl ethylene acetal, benzylidene acetal, 2,2,2-trichloroethylene acetal, etc., cyclic ketals such as isopropylidene ketal, cyclopentylidene Ketal, cyclohexylidene ketal, cycloheptylidene ketal, 1-phenylethylene ketal, 2,4-dimethoxybenzylidene ketal, etc., and cyclic orthoesters such as methoxy oxymethylene acetal, ethoxymethylene acetal and so on.

Preferred examples of R ³ , R ⁴ , R ⁵ and R ⁶ are hydroxyl groups, respectively.

An example of a preferred structure of the compound of formula (I) is the stereostructure represented by formula (V).

The method for producing the above-mentioned compound of the present invention is explained below.

[其中各符号定义如上]或其盐，或其反应性衍生物，与式(III)化合物[其中各符号定义如上]或其盐进行反应。The compound (I) or salt of the present invention is typically prepared by the following method: making the carboxylic acid of formula (II)

[wherein each symbol is as defined above] or a salt thereof, or a reactive derivative thereof, and a compound of formula (III) [wherein each symbol is as defined above] or a salt thereof is reacted.

Reactive derivatives of the carboxylic acids used in the above reactions can be prepared by, for example, the acid halide method, the azide method, the mixed anhydride method (useful 'pairing acids' include isobutoxycarbonyl chloride, pivaloyl chloride, etc.), symmetrical anhydride method, method using a fusing agent such as N,N'-carbodiimidazole, N,N'-dicyclohexylcarbodiimide, N,N'-diisopropyl Carbodiimide, 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide, N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline, Diethyl phosphocyanate, diphenylphosphoryl azide, 2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethyluronium tetrafluoroborate, hexafluoro 2-(1H-Benzotriazol-1-yl)-1,1,3,3-tetramethyluronium phosphate, benzotriazol-1-yloxytris(dimethylamino)phosphonium hexafluorophosphate Onium, benzotriazol-1-yloxytripyrrolidinylphosphonium hexafluorophosphate, bromotripyrrolidinylphosphonium hexafluorophosphate, 2-(5-norbornene-2,3-difluoroborate Formamido)tetramethyluronium or similar, in 4-dimethylaminopyridine, 3-hydroxy-3,4-dihydro-4-oxo-1,2,3-benzotriazole, N- A method using the above-mentioned condensing agents in the presence of hydroxysuccinimide, N-hydroxy-5-norbornene-2,3-dicarboxamide, 1-hydroxybenzotriazole or the like, or a method using their active esters .

The above reaction is generally carried out using 0.5-10 molar equivalents of compound (III) relative to compound (II) in a solvent. Usable solvents include aromatic hydrocarbons, such as benzene, toluene, xylene, etc.; halogenated hydrocarbons, such as methylene chloride, chloroform, etc.; saturated hydrocarbons, such as hexane, heptane, cyclohexane, etc.; ethers, such as diethyl ether , tetrahydrofuran, dioxane, etc.; nitriles, such as acetonitrile, etc.; sulfoxides, such as dimethyl sulfoxide; amides, such as N, N-dimethylformamide, etc.; water. These solvents may be used alone or in combination of two or more, for example, in a ratio of 1:1 to 1:10. The reaction temperature is usually about -80 to 100°C, preferably -50 to 50°C. The reaction time may be about 1 to 96 hours, preferably about 1 to 72 hours.

Compounds of formula (II) or salts thereof are typically synthesized by methods disclosed in Acta Chemica Scandinavica B36, 515-518 (1982).

Compounds of formula (III) can be synthesized by known methods. Commercially available products can also be used.

Compound (I) or a salt of the present invention can be synthesized by, for example, introducing a substituent into the amino group of compound (I) (wherein R ¹ =NH ₂ ). Substituents may, for example, be derived from carboxylic acids or salts or reactive derivatives thereof.

The compound (I) or its salt of the present invention can be prepared by combining the compound of formula (IV) [wherein each symbol is as defined above] or its salt or its reactive derivative with the compound of the following formula: R ⁹ -X [wherein R ⁹ represents A substituted acyl group and a hydrocarbyl group while X represents a leaving group] or a salt thereof or a reactive derivative thereof is prepared by reacting.

The "acyl" of R ⁹ includes the same 'acyl' as the aforementioned 'acyl' of the amido group of R ¹ . A preferred example of the acyl group at ^R is any of the aforementioned protectable α-L-amino acid residues or any sequence of two or more amino acid residues, more preferably the protectable L-valyl-L- Leucyl and protectable (S)-2-amino-4-pentenoyl.

The "hydrocarbyl" of ^R9 includes the same "hydrocarbyl" as the "hydrocarbyl" of the aforementioned optionally substituted hydrocarbyl-substituted amino group of ^R1 .

Examples of leaving groups for X are hydroxyl, halogen atoms (for example, fluorine, chlorine, bromine, especially chlorine), azido, C _1-20 acyloxy, C _1-6 alkoxy (for example methoxy group, ethoxy, propoxy, isopropoxy, butoxy, etc.), C _6-10 aryloxy group (for example, phenoxy, 1-naphthyloxy, 2-naphthyloxy, etc.) , C _7-11 aralkyloxy (for example, benzyloxy, etc.) and the like.

C _1-20 acyloxy as the leaving group of X includes, for example, C _1-20 alkanoyloxy, preferably C _1-6 alkanoyloxy (such as formyloxy, acetyloxy, propionyloxy , isopropionyloxy, butyryloxy, pentanoyloxy, hexanoyloxy, heptanoyloxy, octanoyloxy, nonanoyloxy, etc.), C _7-16 aroyloxy (for example benzoyloxy, 1-naphthoyloxy, 2-naphthoyloxy, etc.), 5- or 6-membered heterocyclylcarbonyloxy or fused heterocyclylcarbonyloxy (for example, 3-pyrrole Carbonyloxy, 1-imidazolecarbonyloxy, 1-pyrazolecarbonyloxy, 3-isothiazolecarbonyloxy, 3-isoxazolecarbonyloxy, pyrazinecarbonyloxy, 2-pyrimidinecarbonyloxy, 3 -pyrazinecarbonyloxy, 2-indenylcarbonyloxy, 2-isoindolecarbonyloxy, 1-indolecarbonyloxy, 2-furoyloxy, 2-thienoyloxy, Nicotinyloxy, isonicotinoyloxy, morpholinocarbonyloxy, piperidinocarbonyloxy, piperazinylcarbonyloxy, etc.), carbamoyloxy, mono-substituted carbamoyloxy or a di-substituted carbamoyloxy group (wherein the substituents include the same substituents as the above-mentioned "substitutable carbamoyl" for the acyl group of the amido group of ^R ), a thiocarbamoyloxy group, a -substituted thiocarbamoyloxy or di-substituted thiocarbamoyloxy (wherein the substituents include the same as the above-mentioned "substitutable thiocarbamoyl" for the acyl group of the amido group of ^R substituent), C _1-20 alkylsulfonyloxy, especially C _1-6 alkylsulfonyloxy (such as methylsulfonyloxy, ethylsulfonyloxy, propylsulfonyloxy, etc. etc.), C _6-14 arylsulfonyloxy groups which may be substituted by one or more C _1-3 alkyl groups (for example, benzenesulfonyloxy, 1-naphthylsulfonyloxy, 2-naphthylsulfonyl acyloxy, tosyloxy, etc.), sulfamoyloxy, mono-substituted sulfamoyloxy or di-substituted sulfamoyloxy (wherein the substituent includes the ^amide The same substituent as the "substitutable sulfamoyl" of the acyl group), C _1-20 alkoxycarbonyloxy, especially C _1-6 alkoxycarbonyloxy (for example, methoxycarbonyloxy , ethoxycarbonyloxy, propoxycarbonyloxy, etc.), C _6-14 aryloxycarbonyloxy (for example, phenoxycarbonyloxy, 1-naphthyloxycarbonyloxy, 2-naphthalene oxycarbonyloxy, etc.) and the like.

Preferable examples of the leaving group of X are hydroxyl group, halogen atom, azido group, C _1-20 acyloxy group, more preferably hydroxyl group and halogen atom.

The conditions of this reaction and the method of synthesizing the carboxylic acid derivative can be, for example, the same as the aforementioned reaction between compound (II) and compound (III).

Regardless of the R ^a in formula R ^a -R ^b -R ^c , R ^b , and the bond between each unit of R ^c is an amide bond or an ester bond, the carboxyl functional group of R ^d (its other functional groups can be properly protected in advance ) can be activated and condensed with a matching amine or alcohol compound (also suitably previously protected). This condensation product is partially purified and partially deprotected, if necessary. Then, the product was further subjected to a similar condensation reaction. Or when the condensation product is a protected final product, removal of all protecting groups and, if necessary, purification of the product yields a purified final product.

The following protecting groups can be used to protect amino, carboxyl, hydroxyl, and carbonyl groups used in the above series of synthetic reactions.

Amino protecting groups that can be used include amide-forming protecting groups such as formyl, acetyl, chloroacetyl, dichloroacetyl, trichloroacetyl, trifluoroacetyl, acetoacetyl, o-nitrophenylacetyl etc.; carbamate-forming protecting groups such as tert-butoxycarbonyl, benzyloxycarbonyl, p-methoxybenzyloxycarbonyl, p-nitrobenzyloxycarbonyl, 2-chlorobenzyloxycarbonyl, 2,4 -Dichlorobenzyloxycarbonyl, dibenzyloxycarbonyl, 2,2,2-trichloroethoxycarbonyl, 2-trimethylsilylethoxycarbonyl, 1-methyl-1-(4- Diphenyl)ethoxycarbonyl, 9-fluorenylmethoxycarbonyl, 9-anthracenylmethoxycarbonyl, isonicotinoyloxycarbonyl, 1-adamantyloxycarbonyl, etc.; trityl, and phthaloyl.

Hydroxyl protecting groups that can be used include ether-forming protecting groups such as methoxymethyl, benzyloxymethyl, tert-butoxymethyl, 2-methoxyethoxymethyl, 2-(trimethylmethoxy Silyl)ethoxymethyl, methylthiomethyl, 2-tetrahydropyranyl, 4-methoxy-4-tetrahydropyranyl, 2-tetrahydropyranyl, benzyl, p-methyl Oxybenzyl, p-nitrobenzyl, o-nitrobenzyl, 2,6-dichlorobenzyl, trityl, etc.; silyl ether protecting groups, such as trimethylsilyl, three Ethylsilyl, triisopropylsilyl, isopropyldimethylsilyl, diethylisopropylsilyl, tert-butyldimethylsilyl, tert-butyldiphenyl Silyl, tribenzylsilyl, triphenylsilyl, methyldiphenylsilyl, etc.; and ester-forming protecting groups such as formyl, acetyl, chloroacetyl, dichloroacetyl , Trichloroacetyl, pivaloyl, benzoyl, benzyloxycarbonyl, 2-bromobenzyloxycarbonyl, etc.

Carboxy protecting groups that can be used include ester-forming protecting groups such as methyl, ethyl, methoxymethyl, methoxyethoxymethyl, benzyloxymethyl, tert-butyl, benzyl, p-methoxy phenylbenzyl, p-nitrobenzyl, o-nitrobenzyl, benzhydryl, trityl, 2,2,2-trichloroethyl, 2-trimethylsilylethyl, allyl Group, cyclohexyl, cyclopentyl, phenacyl, etc.; silyl ester protecting group, such as trimethylsilyl, triethylsilyl, tert-butyldimethylsilyl, Isopropyldimethylsilyl, dimethylphenylsilyl and the like.

Carbonyl protecting groups include those that form acetal-, ketal-, dithioacetal-, or dithioketal, such as dimethyl, diethyl, dibenzyl, diacetyl, etc.; Substituting the protecting group of 1,3-dioxane or 1,3-dioxolane, forming the protecting group of 1,3-dithiane or 1,3-dithiolane, and forming N,N-dimethyl Protective groups for 2,4-dinitrophenyl and other substituted hydrazones.

Techniques for removing these amino-protecting groups, hydroxyl-protecting groups, carbonyl-protecting groups, and carboxyl-protecting groups include methods using acids, methods using bases, reduction methods, UV methods, hydrazine methods, phenylhydrazine methods, N-methyl disulfide Sodium carbamate, tetrabutylammonium fluoride method, palladium acetate method, mercuric chloride method and Lewis acid method. Those conventional methods and/or other known methods may optionally be used.

The method using an acid is a common method for the hydrolysis of amides, esters, silyl esters or silyl ethers and is suitable for removal of the corresponding class of protecting groups. For example, this method is generally used for tert-butoxycarbonyl, p-methoxybenzyloxycarbonyl, benzyloxycarbonyl, 9-anthracenylmethoxycarbonyl, 1-methyl-1-(4-diphenyl base) ethoxycarbonyl, adamantyloxycarbonyl, or trityl protected amino deprotection, and with methoxymethyl, tert-butoxymethyl, 2-tetrahydropyranyl, 4- Deprotection of methoxy-4-tetrahydropyranyl, 2-tetrahydrofuranyl or trityl protected hydroxy groups. Preferable acids include organic acids such as formic acid, trifluoroacetic acid, benzenesulfonic acid, p-toluenesulfonic acid and the like and inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid and the like.

Like the above-mentioned method using an acid, the method using a base is an ordinary method for hydrolyzing an amide, ester or the like bond, and is suitable for removing the corresponding kind of protecting group. For example, it is advantageous to remove an amino group protected with a 9-fluorenylmethoxycarbonyl group with an organic base. Preferred bases include such inorganic bases: alkali metal hydroxides, such as lithium hydroxide, sodium hydroxide, potassium hydroxide, etc.; alkaline earth metal hydroxides, such as magnesium hydroxide, calcium hydroxide, etc.; alkali metal carbon acid salts, such as sodium carbonate, potassium carbonate, etc.; alkaline earth metal carbonates, such as magnesium carbonate, calcium carbonate, etc.; alkali metal bicarbonates, such as sodium bicarbonate, potassium bicarbonate, etc.; alkali metal acetates, For example, sodium acetate, potassium acetate, etc.; alkaline earth metal phosphates, such as calcium phosphate, magnesium phosphate, etc.; alkali metal hydrogen phosphates, such as disodium hydrogenphosphate, dipotassium hydrogenphosphate, etc.; and ammonia water; and such organic bases : Trimethylamine, triethylamine, diisopropylethylamine, pyridine, picoline, N-methylpyrrolidine, piperidine, N-methylpiperidine, N-methylmorpholine, 1,5-di Azabicyclo[4.3.0]non-5-ene, 1,4-diazabicyclo[2.2.2.]octane, 1,8-diazabicyclo[5.4.0]-7-undecane ene and so on.

Generally, trichloroacetyl, trifluoroacetyl, o-nitrophenylacetyl, 2,2,2-trichloroethoxycarbonyl, benzyloxycarbonyl, p-nitrobenzyloxycarbonyl, 2,4- Amino groups protected by dichlorobenzyloxycarbonyl, isonicotinoyloxycarbonyl, trityl, etc.; hydroxy groups protected by benzyl, p-nitrobenzyl, etc.; and benzyloxymethyl, Benzyl, p-nitrobenzyl, phenacyl, 2,2,2-trichloroethyl, benzhydryl and other protected carboxyl groups. Preferred means of reduction include reduction with sodium borohydride, reduction with zinc/acetic acid, and catalytic reduction.

The ultraviolet method is generally used to remove the hydroxyl or carboxyl group protected with o-nitrobenzyl group.

The hydrazine method is generally used to remove amino groups protected with phthaloyl (eg, phthalimide).

The phenylhydrazine method is generally used to remove amino groups protected with acetoacetyl groups.

The sodium N-methyldithiocarbamate method is generally used to remove chloroacetyl-protected amino or hydroxyl groups.

The tetrabutylammonium fluoride method is generally used to remove 2-trimethylsilylethyl carbamate, silyl ether or silyl ester in order to regenerate amino, hydroxyl or carboxyl groups according to the situation.

The palladium acetate method is generally used to remove allyl esters to regenerate carboxyl groups.

The mercuric chloride method is generally used to remove hydroxyl groups protected with methylthiomethyl.

The Lewis acid method is generally used to remove hydroxyl groups protected with 2-methoxyethoxymethyl groups. Among other compounds, preferred Lewis acids include zinc bromide and titanium tetrachloride.

The intermediates, reaction products and target products produced by the above reaction series can be separated and purified according to known purification procedures or similar procedures, such as concentration, concentration under reduced pressure, solvent extraction, crystallization, recrystallization, recrystallization, etc. Partition and chromatographic purification.

As another alternative production method, the compound of the present invention [for example, the compound of the above paragraph (12), (13), (14), (15) and (16)] can be produced by a microbial method.

The microorganisms that can be used to implement the production method of the present invention include bacillus HC-70 (hereinafter sometimes referred to as strain HC-70) isolated from Japan NaraPrefecture soil and abnormal bacillus HC-72 isolated from Japan Aichi Prefecture soil ( Hereinafter, it may be referred to as strain HC-72).

The taxonomic investigation of the strain HC-70 according to conventional methods revealed that the microorganism was a Gram-positive mobile facultative anaerobic bacillus with a cell size of 1.3-1.4 microns x 3.0-4.2 microns. The formation of endospores was observed. The spores are ellipsoidal and the location of the spores is in the center of the cell; no swollen cysts are observed. As a chemotaxonomic characteristic of this species, the cell wall diaminopimelic acid is mesodiaminopimelic acid (endo rac-DAP), the main menaquinone is menaquinone-7 (MK-7), and the G+C content of DNA is 35.0 mol%. The major cellular fatty acids are iso-C _15:0 , C _16:0 anteiso-C _17:0 . According to the classification criteria given in Bergey's "Bergey' Manual of Systematic Bacteriology" Volume II, the species is Bacillus sp. This strain showed adequate growth on broth agar and hydrolyzed casein, gelatin and starch.

A taxonomic investigation of the species HC-72 in an ordinary manner revealed that the species is a Gram-positive mobile aerobic bacillus with a cell size of 1.3 x 3.0-4.2 microns. Endospores were ellipsoidal and the location of the spore was in the center of the cell; no swollen sporangia were observed. As a chemotaxonomic property of this species, meso-diaminopimelic acid (meso-DAP) was not observed as its cell wall component, and the isoprenoid quinone was menaquinone-7 (MK-7) , and the G+C content of DNA is 36.8 mol%. The main cellular fatty acids are iso-C _15:0 , anteiso-C _16:1 , anteiso-C _17:1 . According to the classification criteria given in Bergey's "Bergey' Manual of Systematic Bacteriology" Volume II, the species is Bacillus sp. In addition, none of acid formation from sugars, gelatin hydrolysis, and sodium chloride requirements were found. Therefore, this microorganism was identified as Bacillus anomaly.

Above-mentioned bacillus HC-70 has been deposited in Institute for Fermentation, Osaka (IFO) on June 20, 1997, and the accession number is IFO 16098. In addition, this microorganism has been deposited at the National Institute of Bioscience and Human Technology (NIBH, 1-3, Higashi 1-chome, Yatabe-cho, Tsukuba-shi, Ibaraki, Japan) on July 2, 1997 under the accession number FERM BP -6001.

The above-mentioned bacillus aneurysm HC-72 was deposited on June 1, 1998 under the accession number IFO16179. Furthermore, this microorganism was deposited with NIBH on July 8, 1998 under accession number FERM BP-6385.

As a general characteristic of microorganisms, Bacillus can also undergo mutations whether spontaneously or through mutagenesis. Thus, for example, obtained by irradiation with X-rays, gamma-rays or ultraviolet rays, or by treatment with different mutagenic substances, grown on media containing different mutagenic substances, or obtained by other means Mutants as well as spontaneous mutants can be used in the present invention, unless they are completely unable to synthesize substances related to HC-70 (for example, HC-70I, HC-70II, HC-70III, HC-70I-A, HC-70I- B, etc.).

The medium used in the method of the present invention may be liquid or solid as long as it contains nutrients that the specific strain can use, but liquid medium is preferred for mass production. Assimilable carbon sources, digestible nitrogen sources, inorganic substances and micronutrients are added to the medium. Carbon sources include, but are not limited to, glucose, lactose, sucrose, maltose, dextrin, starch, glycerol, mannitol, sorbitol, inositol, oils and fats (e.g., soybean oil, olive oil, rice bran oil, sesame oil, pork oil, chicken fat, etc.). Nitrogen sources include, but are not limited to, meat extracts, yeast extracts, dried yeast, soybean meal, corn steep liquor, peptone, cottonseed meal, sucrose honey, urea, and ammonium salts (e.g., ammonium sulfate, ammonium chloride, nitric acid ammonium, ammonium acetate, etc.). In addition, salts of sodium, potassium, calcium, magnesium, etc., salts of other metals such as iron, manganese, zinc, cobalt, nickel, etc., salts of phosphoric acid, boric acid, etc., and salts of organic acids such as acetic acid, propionic acid, etc. Appropriate amount can be selected after selection. In addition, amino acids (eg, glutamic acid, aspartic acid, alanine, lysine, valine, methionine, proline, etc.), vitamins (eg, vitamins B ₁ and B ₂ , niacin, vitamin B ₁₂ and C, etc.), and nucleic acids (for example, purine nucleotides, pyrimidine nucleotides and derivatives thereof) can also be added. Of course, for adjusting the pH of the medium, mineral or organic acids, bases, and/or buffers may be added. As an antifoaming agent, an appropriate amount of oil or surfactant can also be added.

A suitable culture method may be selected from fixed culture, shaking culture and aerated submerged culture. Of course, for bulk treatments, so-called aerated submerged cultures are preferred.

Of course, the culture conditions depend on the type and composition of the medium used, the specific bacterial species and the culture method selected, but the preferred incubation temperature is generally 15-37° C., and the initial pH value is about 5-9. In particular, the mid-culture temperature is preferably 20-30°C and the initial pH is preferably about 6-8. The incubation time also depends on the above conditions, but the incubation should be continued until the accumulation of the compound of interest reaches a maximum level. For shaking or aeration culture in liquid medium, the required incubation time is generally about 1-10 days.

Under the above culture conditions, the compounds HC-70I, II, and III described below were produced and accumulated, and then the respective compounds were extracted and purified according to their specific chemical properties. The HC-70I, II and III of interest can be collected from the culture fluid by suitable techniques selected from methods generally used for the collection of microbial metabolites from the culture fluid. For example, since HC-70I, II and III are water-soluble amphiphilic compounds and are mostly present in the culture supernatant, cells are first removed from the broth by filtration or centrifugation.

The culture supernatant thus isolated can be further purified by known chromatographic methods to obtain pure HC-70I, II and III. Chromatographic stationary phases that can be used for this purpose include those utilizing stationary phases having different adsorption affinities for substrate compounds, such as activated carbon [e.g., activated carbon for chromatography, granular carbon "Shirasagi" (Takeda Chemical Industry Co., Ltd. company), etc.], adsorbent resin [for example, DIAION HP-20, HP-20S or HP-20SS, SEPABEADS SP-207 or SP-850 (Mitsubishii Chemical Industry Co., Ltd.), Amberlite XAD-I or XAD-II (Rohm & Haas Company, USA) etc.], microcrystalline cellulose [for example, Avicel (Asahi Chemical Industry Co., Ltd.), Funacel (Funakoshi Co., Ltd.) etc.], and silica gel [for example, Kieselguhr 60 (Merck &. Co. , Germany) etc.]; those utilizing specific functional groups for stationary phases such as cation exchange resins [eg, Amberlite IR-120, IRC-50, or CG-50 (Rohm & Haas, USA), Dowex 50W (Dow Chemical company, USA), DIAION PK-216 or UBK-510L (Mitsubishii Chemical Industry Co., Ltd.), and CNP-80 (Bayer, Germany) etc.], anion exchange resin [for example, Amberlite IRA-402, IRA-67, or IRA-68 (Rohm & Haas Company, USA), Dowex 1 (Dow Chemical Company, USA), DIAION SA-21A, PA-406, PA-412 or WA-30 (Mitsubishii Chemical Industry Co., Ltd.), etc.], ion Exchange cellulose [CM-Cellulose (Pharmacia, Sweden) etc.], ion-exchange Sephadex [for example, QAE-Sephadex or CM-Sephadex (Pharmacia, Sweden) etc.]; and those stationary phases that take advantage of molecular weight differences, such as molecular sieves [eg, Sephadex G10 or LH-20 (Pharmacia, Sweden), etc.].

The solvent that is used as chromatographic mobile phase depends on the kind and the characteristic of stationary phase, for example can be any following solvent or suitable solvent mixture, the example of described solvent is water, alkali (such as sodium hydroxide, potassium hydroxide, carbonic acid Sodium hydrogen, ammonia, etc.) aqueous solution, acid (such as hydrochloric acid, sulfuric acid, acetic acid, formic acid, phosphoric acid, etc.) aqueous solution, saline solution (such as sodium chloride solution, acetate buffer, phosphate buffer, etc. etc.) and water-miscible organic solvents (for example, methanol, ethanol, isopropanol, acetone, acetonitrile, etc.).

Preparative high performance liquid chromatography (HPLC) may also be advantageously employed in the practice of the present invention in order to purify the compound of interest. When this method is used, the stationary phase is preferably one of octadecylsilane (ODS) series, polymer series or silica gel series. As the ODS series stationary phase, for example, YMC gel (YMC) or TSK gel (Tosoh) can typically be mentioned. As a polymer series stationary phase, for example, you can choose ODP (Asahi Chemical Industry Co., Ltd.), which is an octadecylated polymer; or NH2P (Asahi Chemical Industry Co., Ltd.), which is a polyamine-modified polymer. things. As the mobile phase, water, acidic aqueous solution, saline aqueous solution, methanol, acetonitrile and the like can be used alone or as a suitable mixture.

Crystallization is also a useful technique for purifying the compounds of interest of the present invention. As the crystallization solvent, water, methanol, ethanol, isopropanol, acetone, acetonitrile and the like can be used alone or as a suitable mixture.

HC-70I-A and HC-70I-B can also be obtained and purified from the culture (fermentation) liquid in the same manner as above.

The physical and chemical properties of HC-70I, II and III obtained according to Examples 1 and 2 below are as follows. These compounds are sometimes defined as Compound 1, Compound 2 and Compound 3, respectively.

HC-70I (compound 1)

1) Appearance: colorless crystal

2) Optical rotation: -89° (c=0.53, 0.1N HCl, 24°C)

3) Molecular weight: FAB-MS m/z 654(M+H) ⁺

4) Elemental analysis: (%) (calculated with 1 mole of water)

Found values: C, 55.23; H, 8.03; N, 10.47

Analytical values: C, 55.43; H, 7.95; N, 10.42

5) Molecular formula: C ₃₁ H ₅₁ N ₅ O ₁₀

6) UV spectrum: _λmax (ε)

In water, 258 nm (310)

7) Infrared spectrum: KBr: main absorption peak (wave number, cm ^-1 ):

3300, 2960, 1640, 1540, 1400, 1050, 7008) ¹³ C-NMR spectrum: DMSO-d ₆ , chemical shift (75MHz, δppm; Figure 1)

172.8, 172.6, 172.4, 172.3, 170.7, 142.6, 128.0,

126.5, 126.5, 71.0, 70.9, 67.2, 60.5, 59.1, 57.2,

51.1, 50.9, 49.0, 41.1, 40.5, 30.9, 30.7, 24.0,

23.0, 21.3, 19.3, 19.1, 18.1, 16.89) Amino acid analysis: after hydrolysis with 6N hydrochloric acid at 110°C for 72 hours

Leucine (1 mole), valine (2 moles) 10) color reaction:

Positive: Ninhydrin, Greig-Lieback

Negative: Ehrlich (Indole Test Reagent), Sakaguchi 11) High Performance Liquid Chromatography (HPLC):

Column: YMC-Pack ODS-A, A-312, 150×6.0 mm (YMC)

Mobile phase: 15% (v/v) acetonitrile/0.02M

Phosphate buffer (pH4.5)

Flow rate: 1.0ml/min

Detection: Ultraviolet absorption method, 214 nm

Retention time: 16.8 minutes 12) Thin layer chromatography (TLC):

Stationary phase: silica gel 60F ₂₅₄ , 0.25 mm (Merck, Germany)

Developing solvent: n-butanol/acetic acid/water (12:3:5)

Rf: 0.45HC-70II (Compound 2) 1) Appearance: Colorless crystals 2) Optical rotation: -69° (c=0.50, 0.1N HCl, 24°C) 3) Molecular weight: FAB-MS m/z 555 (M +H) ⁺ 4) Elemental analysis: (%) (calculated with 3 moles of water)

Found values: C, 51.44; H, 7.84; N, 9.32

Analytical values: C, 51.30; H, 7.95; N, 9.20

5) Molecular formula: C ₂₆ H ₄₂ N ₄ O ₉

6) UV spectrum: _λmax (ε)

In water, 257 nm (270)

7) Infrared spectrum: KBr: main absorption peak (wave number, cm ^-1 ):

3370, 2970, 2940, 1680, 1630, 1520, 1400, 1050, 690

8) ¹³ C-NMR spectrum: DMSO-d ₆ /trifluoroacetic acid (9:1), chemical shift (75MHz, δppm; Figure 2)

173.3, 173.0, 172.7, 168.3, 142.6, 128.7, 127.4,

127.1, 71.9, 71.5, 68.1, 61.2, 58.0, 52.0, 49.5,

41.5, 40.8, 30.5, 24.6, 23.4, 21.9, 18.8, 17.9

9) Amino acid analysis: after hydrolysis with 6N hydrochloric acid at 110°C for 24 hours

Leucine (1 mole), Valine (1 mole)

10) Color reaction:

Positive: Ninhydrin, Greig-Lieback

Negative: Ehrlich, Sakaguchi

11) High performance liquid chromatography (HPLC):

Column: YMC-Pack ODS-A, A-312, 150×6.0 mm (YMC)

Mobile phase: 15% (v/v) acetonitrile/0.02M

Phosphate buffer (pH4.5)

 Flow rate: 1.0ml/min

Detection: UV absorption method, 214 nm

  Retention time: 8.1 minutes

12) Thin layer chromatography (TLC):

Stationary phase: silica gel 60F ₂₅₄ , 0.25 mm (Merck, Germany)

Developing agent: n-butanol/acetic acid/water (12:3:5)

Rf: 0.41

HC-70III (compound 3)

1) Appearance: colorless crystals 2) Optical rotation: -67° (c=0.55, 0.1N HCl, 24°C) 3) Molecular weight: FAB-MS m/z 456 (M+H) ⁺ 4) Elemental analysis: ( %) (calculated with 1 mole of water)

Found values: C, 53.14; H, 7.14; N, 8.98

Analytical value: C, 53.27; H, 7.45; N, 8.875) Molecular formula: C ₂₁ H ₃₃ N ₃ O ₈ 6) UV spectrum: _λmax (ε)

In water, 257 nm (350) 7) infrared spectrum: KBr: main absorption peak (wave number, cm ^-1 ):

3390, 2970, 2930, 1660, 1540, 1400, 1070, 1050, 7008) ¹³ C-NMR spectrum: DMSO-d ₆ , chemical shift (75MHz, δppm; Figure 3)

174.3, 172.9, 172.4, 142.7, 127.9, 126.5, 71.2,

70.8, 67.6, 60.9, 52.3, 51.2, 49.0, 42.8, 41.3,

23.9, 23.0, 21.79) Amino acid analysis: after hydrolysis with 6N hydrochloric acid at 110°C for 24 hours

Leucine (1 mole) 10) color reaction:

Positive: Ninhydrin, Greig-Lieback

Negative: Ehrlich, Sakaguchi 11) High Performance Liquid Chromatography (HPLC):

Column: YMC-Pack ODS-A, A-312, 150×6.0 mm (YMC)

Mobile phase: 15% (v/v) acetonitrile/0.02M

Phosphate buffer (pH4.5)

Flow rate: 1.0ml/min

Detection: Ultraviolet absorption method, 214 nm

Retention time: 6.0 minutes 12) thin layer chromatography (TLC):

Stationary phase: silica gel 60F ₂₅₄ , 0.25 mm (Merck, Germany)

Developing solvent: n-butanol/acetic acid/water (12:3:5)

Rf: 0.35HC-70 The chemical formulas of I, II and III are as follows. Compound No. R ¹

Compound 1 is (S)-3-[(2S,3R,4R,5S)-2,3,4,6-tetrahydroxy-5-(L-valyl-L-valyl-L-leucine Acyl)aminocaproyl]amino-3-phenylpropanoic acid (HC-70I, the compound of Example 2).

Compound 2 is (S)-3-[(2S,3R,4R,5S)-2,3,4,6-tetrahydroxy-5-(L-valyl-L-leucyl)aminocaproyl] Amino-3-phenylpropionic acid (HC-70II, compound of Example 1).

Compound 3 is (S)-3-[(2S,3R,4R,5S)-2,3,4,6-tetrahydroxy-5-(L-leucyl)aminocaproyl]amino-3-phenyl Propionic acid (HC-70III, compound of Example 1).

A compound wherein R ¹ =NH ₂ or a salt thereof can be produced by allowing an enzyme to act on Compound 1, 2, or 3 or a salt thereof. To this end, enzymes that can be used include, but are not limited to, exopeptidases (eg, leucine aminopeptidase) and proteases [eg, Actinase E (Kaken Pharmaceutical Co., Ltd.)].

The reaction is generally carried out in water, and for pH control, inorganic or organic acids, bases or buffers can be added. The reaction temperature is not particularly critical unless the reaction of the enzyme is hindered, but it is generally about 10-50°C, preferably 20-40°C. The reaction time depends on the kind and amount of the enzyme, the reaction temperature, and the pH of the solution, but is generally several minutes to several hours.

The chemical formula of the resulting compound (compound 5) wherein R ¹ =NH ₂ is as follows:

Compound 5 is (S)-3-[(2S, 3R, 4R, 5S)-5-amino-2,3,4,6-tetrahydroxyhexanoyl]amino-3-phenylpropionic acid (instance 4 compound).

Since compound 5 is produced and accumulated in the fermentation broth of Bacillus HC-70 and Bacillus aneurysm HC-72, it can also be obtained and purified from the fermentation broth by the same method as above.

The compound of general formula (I) or a salt thereof is sometimes referred to as compound (I) below.

The salts of the compounds of formula (I) of the present invention include pharmaceutically acceptable base addition salts and acid addition salts. Base addition salts include, but are not limited to, alkali metal (eg, sodium, potassium, etc.) salts and alkaline earth metal (eg, calcium, magnesium, etc.) salts. Acid addition salts include, but are not limited to, those with inorganic acids (e.g., hydrochloric, hydrobromic, hydroiodic, sulfuric, phosphoric, etc.) and with organic acids (e.g., acetic, propionic, lactic, succinic, maleic, acid, tartaric acid, citric acid, gluconic acid, ascorbic acid, benzoic acid, methanesulfonic acid, p-toluenesulfonic acid, cinnamic acid, fumaric acid, malic acid, oxalic acid, etc.).

Hydrates and anhydrides of compounds of general formula (I) are also within the scope of the present invention. Examples of hydrates are 0.5 hydrate, 1 hydrate, 1.5 hydrate, 2 hydrate, 2.5 hydrate, 3 hydrate, 3.5 hydrate, 4 hydrate and the like.

Compound (I) of the present invention can be isolated and purified by known methods such as solvent extraction, pH change, phase transfer or redistribution, crystallization, recrystallization and chromatographic purification. The starting compound or salt of compound (I) can also be isolated and purified by a similar method, but the reaction mixture containing it can be directly supplied to the intended reaction.

In the case where the compound (I) of the present invention exists as optical isomers, stereoisomers, positional isomers or rotational isomers, these isomers are also within the scope of the present invention and each such isomer Constructs can be provided as single substances by known synthetic methods or fractionation techniques. For example, when the compound of the present invention exists as optical isomers, each isomer obtained by an optical resolution technique of the compound is also within the scope of the present invention.

Optical isomers can be produced by known methods. Specifically, desired optical isomers can be obtained by using optically synthesized intermediates or by optically resolving the product racemic mixture.

The aforementioned optical resolution can be achieved by known techniques such as the fractional recrystallization method, chiral column method and diastereomer method described below.

(1) Fractional recrystallization method

This method involves salting a racemic compound with an optically active compound, separating it by fractional crystallization, and optionally neutralizing it to provide the free optical isomer.

(2) Chiral column method

This method involves applying a racemic compound or a salt thereof to a chiral column. When using liquid chromatography, a typical procedure includes applying a mixture of optical isomers to a chiral column, such as ENANTIO-OVM (Tosoh), and water, a buffer (such as phosphate buffer), an organic solvent (such as ethanol, methanol, acetonitrile, etc.) or a mixture of such solvents to recover the desired optical isomer. When using gas chromatography, the use of a chiral column such as CP-Chirasil DeXCB (G.L. Science) can achieve the desired fractionation.

(3) Diastereomer method

This method involves reacting a racemic mixture with an optically active reagent to prepare a mixture of diastereomers, subjecting the mixture to conventional fractionation (e.g., fractional recrystallization, chromatography, etc.) to provide a single substance, and further subjecting it to chemical Treatment such as hydrolysis to cleave the optically active reagent moiety. For example, when the compound of the present invention has a hydroxyl group or a primary or secondary amino group, by reacting the substrate compound with an optically active organic acid (for example, MTPA [α-methoxy-α-(trifluoromethyl)phenylacetic acid], (-) -Methoxyacetic acid, etc.) can be subjected to condensation reactions to obtain the corresponding ester or amide diastereomers. On the other hand, if the compound of the present invention has a carboxyl group, the amide or ester diastereomers can be obtained by condensation reaction of the substrate compound with an optically active amine or alcohol reagent. The separated diastereomers can then be converted into the optical isomers of the starting compound by the method of acid or base hydrolysis.

Compound (I) of the present invention has low toxicity, has commendable pharmacological activity, for example has very high antibacterial activity to Helicobacter pylori bacterium represented by Helicobacter pylori, so can effectively prevent or treat Helicobacter pylori infection and / or diseases related to ammonium produced by Helicobacter pylori (such as duodenal ulcer, gastric ulcer, gastritis (including chronic gastritis), gastric cancer, gastric MALT lymphoma, hepatic encephalopathy, diabetes mellitus, urticaria).

Therefore, the pharmaceutical composition containing the compound (I) of the present invention can be administered orally or parenterally to humans and other mammals (for example, dogs) as a safe antibacterial agent or as a safe antiulcer drug alone or together with a pharmaceutical carrier. , felines, monkeys, rats, mice, horses, cattle, etc.) administration. Generally, oral administration is preferred.

Dosage forms for oral administration include, but are not limited to, tablets (including dragees and coated tablets), pills, granules, fine granules, powders, capsules (including soft capsules), syrups, emulsions, and suspensions. Dosage forms for parenteral administration include, but are not limited to, injections, infusions, drips and suppositories.

For example, the gastric mucoadhesive composition of the present invention is a composition containing (a) the compound (I) of the present invention (as an active ingredient having, for example, anti-Helicobacter pylori activity), (b) lipid and/or polyglycerol A composition of a fatty acid ester and (c) a viscous agent (a substance which becomes sufficiently viscous to adhere to the gastric mucosa in contact with water). The composition at least adheres itself to the gastric mucosa and/or then stays in the stomach and releases the active ingredients contained therein, such as anti-H. Helicobacter activity).

An example of the above gastric mucoadhesive composition may be a composition containing (a) an anti-H. , preferably the composition further contains (d) a substance that swells the viscous agent (for example, Alcaligenes polysaccharide and/or low-substituted hydroxypropyl cellulose as the swelling substance). Although there is no particular limitation on its dosage form, the composition is preferably a solid composition, especially a matrix-containing composition. For example, the matrix may be a gastric mucoadhesive matrix comprising (a), (b) polyglycerol fatty acid ester and (c), or a gastric mucoadhesive matrix comprising (a), (b) lipid and (c) . A preferred matrix is a gastric mucoadhesive matrix containing (b) polyglycerol fatty acid ester. A preferable example of the gastric adhesive composition of the present invention is a composition further comprising (d) a substance that swells a viscous agent.

The gastric mucoadhesive matrix comprising the four components (a), (b), (c) and/or (d) is preferably a matrix in which the viscous agent is dispersed in a polyglycerol fatty acid ester or lipid-containing In the matrix or a matrix coated with a viscous agent. For example, the melting point of the gastric mucoadhesive matrix may be from about 30°C to about 120°C, preferably from about 40°C to about 120°C.

The polyglycerol fatty acid ester used in the present invention is an ester of polyglycerol with a fatty acid, and may be mono- to polyester (diester, triester, etc.). Polyglycerol fatty acid esters are characterized in that they do not undergo polymorphic transformation or undergo any material reaction with active ingredients, enabling those coexisting ingredients to remain in a non-inactivated state and to be stable for a long period of time.

Polyglycerol is defined as "a molecule containing n (cyclic) to (n+2) (linear or branched) hydroxyl groups and (n-1) (linear or branched) to n (cyclic) ) ether-bonded polyol” [polyglyceride, Sakamoto Yakuhin Kogyo Co., Ltd., published on October 4, 1994], and any straight-chain or branched-chain ester can be used in the present invention.

(其中n代表不低于2的整数的聚合度)。n值一般约2至约50，优选约2至约20，更佳结果约2至约10。For example, compound (I) of the formula can be used

(wherein n represents the polymerization degree of an integer not lower than 2). The value of n is generally about 2 to about 50, preferably about 2 to about 20, more preferably about 2 to about 10.

Polyglycerols include, but are not limited to, diglycerol, triglycerol, tetraglycerol, pentaglycerol, hexaglycerol, heptaglycerol, octaglycerol, nonaglycerol, decaglycerol, fifteen glycerol, eicosylglycerol, and triacylglycerol. Among them, tetraglycerol, hexaglycerol or decaglycerol is used in many cases.

Fatty acids include, but are not limited to, saturated or unsaturated fatty acids each containing from about 8 to about 40, preferably from about 12 to about 25, more preferably from about 15 to about 22 carbon atoms. Preferred fatty acids are stearic acid, oleic acid, lauric acid, linoleic acid, linolenic acid, ricinoleic acid, caprylic acid, capric acid, or behenic acid.

Polyglycerol fatty acid esters include, but are not limited to, hexa(tetra)glycerides of behenate, mono(capric)glycerides of caprylate, diglycerides of caprylic acid, diglycerides of capric acid (di(tri)glycerides) Glycerides, mono(hexa)glycerides of laurate, mono(deca)glycerides of laurate, mono(tetra)glycerides of oleate, mono(hexa)glycerides of oleate, mono(deca)glycerides of oleate, oleic acid Two (three) glycerides, two (tetra)glycerides of oleate, sesqui(ten) glycerides of oleate, five (tetra) glycerides of oleate, five (six) glycerides of oleate, ten (ten) oleic acid Glycerides, Mono(7) Glycerides of Linoleate, Di(3) Glycerides of Linoleate, Di(3) Glycerides of Linoleate, Di(4) Glycerides of Linoleate, Di(6) Linoleate Glycerides, mono (di) glyceryl stearate, mono (tetra) glyceryl stearate, penta (tetra) glyceryl stearate, mono (ten) glyceryl stearate, tri (tetra) stearate Glycerides, penta(hexa)glyceryl stearate, tri(hexa)glyceryl stearate, deca(deca)glyceryl stearate, mono(tetra)glyceryl palmitate, mono(hexa)glyceryl palmitate , mono(dec)glyceryl palmitate, tri(tetra)glyceryl palmitate, tri(hexa)glyceryl palmitate, sesqui(hexa)glycerol palmitate, penta(tetra)glyceryl palmitate, pentaglycerol palmitate (6) Glycerides, deca(dec)glyceryl palmitate, and polyglyceryl polyricinoleate (eg, tetraglyceryl polyricinoleate).

Preferred polyglycerol fatty acid esters include, but are not limited to, for example, hexa(tetra)glycerol behenate (e.g., HB-310 ^™ , Sakamoto Yakuhin Kogyo Co., Ltd.; Poem J-46B ^™ , Riken Vitamin Co.), pentastearate (Tetra)glycerides (for example, PS-310 ^™ , Sakamoto Yakuhin Kogyo Co., Ltd.), mono(tetra)glycerides of stearate (for example, MS-310 ^™ , Sakamoto Yakuhin Kogyo Co., Ltd.), penta(hexa)stearate ) glycerides (for example, PS-500 ^™ , Sakamoto Yakuhin Kogyo Co., Ltd.), mono(deca)glyceryl stearate, polyglycerol polyricinoleate (for example, tetraglycerol polyricinoleate, etc.) (eg CRS-75 ^™ , Sakamoto Yakuhin Kogyo Co., Ltd.) and mixtures of these glycerides.

These polyglycerin fatty acid esters may be used alone or as a mixture of two or more, preferably about 2 or about 3 kinds.

The molecular weight of polyglycerol fatty acid ester is generally about 200 to about 5000, preferably about 300 to about 3000, more preferably about 2000 to about 3000. The polyglycerol fatty acid ester generally has a hydrophilic-lipophilic balance (HLB) value of about 1 to about 22, preferably about 1 to about 15, more preferably about 1 to about 9, more preferably about 1 to about 4. Two or more polyglycerol fatty acid esters different in HLB value from each other may be used in combination to provide a specified HLB value. The release and decomposition kinetics of the active drug substance can be controlled by rationally adjusting the HLB of the polyglycerol fatty acid ester as required.

Refer to the specific active ingredient (for example, anti-H. pylori agent, etc.), viscous agent, swelling substance (for example, Alcaligenes polysaccharide and/or low-substituted hydroxypropyl cellulose, etc.), its specific combination and the purpose of the composition Form, can choose suitable polyglycerol fatty acid ester. However, preference is given to using compounds which are solid at atmospheric temperature (about 15°C). For example, the polyglycerol fatty acid ester may have a melting point of about 15°C to about 80°C, preferably about 30°C to 75°C, more preferably about 45°C to about 75°C.

Select suitable polyglycerol fatty acid esters according to the type of active ingredient used and the dosage form used. In general, polyglycerols having a degree of polymerization of from about 2 to about 16 are preferred. A range of about 2 to about 10 is particularly preferred. The preferred ester should be that the fatty acid forms an ester bond with at least one (polymerization degree +2) hydroxyl group, more preferably one or more fatty acids with not less than about 60% of the total, especially not less than about 80% The hydroxyl groups in glycerol form ester bonds. The fatty acids are preferably saturated acids each containing from about 6 to about 22, more preferably from about 15 to about 25, even more preferably from about 18 to about 22 carbon atoms. The fatty acids involved in forming an ester bond may be of the same kind or different kinds.

In producing a solid composition using a mixture of two or more different polyglycerol fatty acid esters according to the present invention, liquid polyglycerol fatty acid esters may also be included in the mixture as long as the final composition is solid at atmospheric temperature.

When polyglycerol fatty acid ester is used as gastric mucoadhesive matrix, the amount of polyglycerol fatty acid ester is generally about 5 to about 98% (weight), preferably about 20 to about 95% ( weight), more preferably about 40 to about 95% (weight), and relative to the active ingredient in the composition, it can be, for example, about 0.01 to about 15000 times the weight, preferably about 0.1 to about 1000 times the weight, and the better result is about 0.1 to about 100 times its weight.

Lipids useful in the present invention are lipids having a melting point of about 40°C to about 120°C, preferably about 40°C to about 90°C.

Lipids include, but are not limited to, saturated fatty acids (e.g., myristic acid, stearic acid, palmitic acid, behenic acid, etc.) or salts thereof (sodium salts, potassium salts, etc.) having about 14 to about 22 carbon atoms ; higher alcohols having about 16 to about 22 carbon atoms (for example, cetyl alcohol, stearyl alcohol, etc.); fatty acid glycerides, such as monoglycerides, diglycerides, triglycerides, etc. , 1-monostearin, 1-monopalmitin, etc.); oils (for example, castor oil, cottonseed oil, tallow, etc., including corresponding hydrogenated oils); waxes (for example, beeswax, carnauba wax, whale waxes, etc.); hydrocarbons (eg, paraffins, microcrystalline waxes, etc.); and phospholipids (eg, hydrogenated lecithin, etc.). Among these lipids, oils, waxes, _C14-22 saturated fatty acids, _C16-22 higher alcohols and hydrocarbons are preferred. More preferred are hydrogenated cottonseed oil, hydrogenated castor oil, hydrogenated soybean oil, carnauba wax, stearic acid, stearyl alcohol and microcrystalline wax. Most preferred are hydrogenated castor oil or carnauba wax.

When lipid is used as gastric mucoadhesive matrix, the amount of lipid, relative to the total weight of the composition, is generally about 5 to about 98% (weight), preferably about 20 to about 95% (weight), more preferably about 40 to about 95% (weight), and relative to the active ingredient in the composition, it is about 0.01 to about 15000 times the weight, preferably about 0.1 to about 1000 times the weight, more preferably about 0.1 to about 100 times the weight.

The above polyglycerol fatty acid esters and lipids may be used in admixture. For example, a combination of polyglycerol fatty acid esters with paraffin or a combination of polyglyceryl fatty acid esters with hydrogenated oils may be mentioned. Specifically, there may be mentioned those selected from the group consisting of hexa(tetra)glyceryl behenate, penta(tetra)glycerol stearate, penta(tetra)glycerol stearate, polyglycerol polyricinoleate ( For example, a mixture of two, three or more of tetraglycerol polyricinoleate, etc.), carnauba wax, hydrogenated castor oil and microcrystalline wax.

When the gastric mucoadhesive matrix containing a viscous agent in addition to the polyglycerol fatty acid ester and/or lipid is used for the composition of the present invention, the total amount of polyglycerol fatty acid ester and lipid, relative to the composition The total weight is generally about 5 to about 98% (weight), preferably about 20 to about 95% (weight), more preferably about 40 to about 95% (weight), and relative to the active ingredient in the composition, is About 0.01 to about 15000 times by weight, preferably about 0.1 to about 1000 times by weight, more preferably about 0.1 to about 100 times by weight.

Lipids can be added to the matrix containing polyglycerol fatty acid esters. Lipids are pharmacologically acceptable water-insoluble substances that modulate the decomposition kinetics of active ingredients. Lipids include those mentioned above.

When lipids and polyglycerol fatty acid esters are used in combination, the amount of lipids and polyglycerol fatty acid esters only needs to be within the range that does not damage the adhesion to the gastrointestinal tract mucosa, and can be selected within the range of the total amount; The amount of lipid can be about 0.01 to about 1000 times by weight relative to polyglycerol fatty acid ester, preferably about 0.1 to about 200 times by weight, more preferably about 0.1 to about 100 times by weight.

The swelling substance used in the present invention is a substance that swells a viscous agent or accelerates swelling of a viscous agent by water.

In the present invention, any kind of swelling substance can be used as long as it has the above-mentioned properties and can be used medicinally. For example, Alcaligenes polysaccharide and/or low-substituted hydroxypropylcellulose are preferred.

In the gastric mucoadhesive composition of the present invention, the amount of the swelling substance is about 0.5 to about 50% (weight), preferably about 1 to about 40% (weight), more preferably about 1 to about 40% (weight), relative to the total weight of the composition. About 30% by weight.

Alcaligenes polysaccharides used in the present invention are linear water-insoluble polysaccharides (β-1,3-glucans) produced by microorganisms (such as Alcaligenes faecalis subsp. myxa, etc.), which include species such as, For example, Alcaligenes polysaccharides 10C3K, 13140, 12607, 12665, 13127, 13256, 13259 and 13660 [New Food Industry, 20, No. 10, p. 49 (1978)]. Among these and other species of Alcaligenes polysaccharides, those acceptable as pharmaceutical bases or excipients can be used. A preferred example is Alcaligenes polysaccharide N (a food additive).

The amount of Alcaligenes polysaccharide in the gastric mucoadhesive composition of the present invention is about 0.5 to about 50% (weight), preferably about 1 to about 40% (weight), more preferably about 1 with respect to the total weight of the composition. to about 30% by weight.

The low-substituted hydroxypropyl cellulose used in the present invention is a cellulose derivative obtained by substituting some hydroxyl groups of cellulose with hydroxypropoxy groups, and its hydroxypropoxy group content is 5.0 to 16.0% (according to the Japanese Pharmacopoeia 12th edition stipulations in). The aforementioned low-substituted hydroxypropyl cellulose is useful, and those having a hydroxypropoxyl content of 7.0 to 13.0% are particularly preferred (for example, L-HPC ^™ , Shin-Etsu Chemical Co., Ltd.). Therefore, those derivatives whose degree of substitution is within the above-mentioned range and whose particle size varies, such as LH-11 ^™ (Shin-Etsu Chemical Co., Ltd., which has a hydroxypropoxyl content of 10.0 to 12.9%, on a 150-micron sieve, can be used among them. Particle size distribution ≥ 98% and particle size distribution ≤ 0.5% on 180 micron sieve), LH-20 ^TM (Shin-Etsu Chemical Co., Ltd., 13.0-16.0% hydroxyphenoxy content, granulation on 75 micron sieve Size distribution ≥ 90% and particle size distribution ≤ 1.0% on 106 micron sieve), LH-21 (Shin-Etsu Chemical Co., Ltd., hydroxyphenoxy content of 10.0-12.9%, particle size distribution ≥ 75 micron sieve 90% and particle size distribution ≤ 1.0% on 106 micron sieve), LH-22 (Shin-Etsu Chemical Co., Ltd., 7.0-9.9% hydroxyphenoxy content, particle size distribution ≥90% on 75 micron sieve and Particle size distribution ≤ 1.0% on 106 micron sieve), LH-31 (Shin-Etsu Chemical Co., Ltd., hydroxypropoxyl content 10.0-12.9%, average particle size not more than 30 micron, particle size under 45 micron sieve distribution ≥ 50% and particle size distribution ≤ 5.0% on a 75 micron sieve).

Preferably LH-22 or LH-31 is used.

The amount of low-substituted hydroxypropyl cellulose in the gastric mucoadhesive composition of the present invention is about 0.5 to about 50% (weight), preferably about 1 to about 40% (weight), more preferably about 0.5 to about 50% (weight) relative to the total weight of the composition. Best results are from about 1 to about 30% by weight.

In the present invention, any viscous agent can be used as long as it becomes viscous enough in contact with water to attach itself to the gastric mucosa and can be used medicinally. However, preference is given to those materials which swell significantly with water and develop a high viscosity. Therefore, viscous agents include synthetic polymers and natural viscous substances.

Preferred synthetic polymers are those having a 2% aqueous solution viscosity at 20°C of from about 3 to about 50,000 centipoise, preferably from about 10 to about 30,000 centipoise, more preferably from about 15 to about 30,000 centipoise. However, when using basic polymers or acidic polymers that viscosify upon neutralization, preferred polymers have a 0.2% solution viscosity of from about 100 to about 500,000 centipoise after neutralization at 20° C., preferably from about 100 to About 200,000 centipoise, more preferably about 1500 to about 100,000 centipoise.

Viscosity values are determined according to a Brookfield viscometer at about 20°C.

The aforementioned polymers are preferably acidic polymers, including but not limited to polymers containing carboxyl groups or sulfur groups and polymers containing corresponding salts. Particular preference is given to carboxyl group-containing polymers and carboxylate salt-containing polymers.

The carboxyl group-containing polymer (including its salt) is preferably an acrylic homopolymer or copolymer containing acrylic acid or its salt as a monomer unit. Salts include monovalent metal salts such as sodium salts, potassium salts and the like and divalent metal salts such as magnesium salts, calcium salts, ammonium salts and the like.

Acrylic polymers, including their salts, including carboxyl group-containing polymers, have a proportion of about 58 to 63% by weight and a molecular weight of about 20×10 ⁴ to about 600×10 ⁴ , preferably about 100×10 ⁴ to about 600×10 ⁴ , more preferably about 100×10 ⁴ to about 500×10 ⁴ . Preferred acrylic polymers, including salts thereof, include homopolymers of acrylic acid and salts thereof. Such polymers are listed under the heading of carboxyvinyl polymers in Japanese Standards for Pharmaceutical Ingredients (October 1998).

Specific examples of the acrylic polymer include carbomer [Carbopol ^™ (hereinafter referred to as Carbopol), The BFGoodrich Company] 940, 934, 934P, 941, 1342, 974P, 971P (NF XVIII), EX ₂ 14 etc., HIVISWAKO ^TM 103, 104, 105, 204 (Wako Pure Chemical Company), NOVEON AA1 ^TM (The BF Goodrich Company) and polycarbophil calcium (USP XXIII)).

Natural viscose agents include, but are not limited to, mucin, agar, gelatin, pectin, carrageenan, sodium alginate, locust bean gum, xanthan gum, tragacanth gum, pullulan, chitosan, waxy starch , sucralfate, Alcaligenes polysaccharide, and cellulose and its derivatives (cellulose sulfate, preferably hydroxypropylcellulose or hydroxypropylmethylcellulose).

The most preferred tackifiers are acrylic polymers or salts thereof.

These tackifiers can be used alone or in combination.

Regarding the amount of viscous agent used in the composition of the present invention, its amount in the gastric mucoadhesive matrix can be, for example, from about 0.005 to about 99% by weight, preferably from about 0.5 to about 45% by weight, more preferably about 1 to about 30% by weight, especially about 1 to about 25% by weight, and more preferably about 1 to about 20% by weight. For example, when the viscous agent is dispersed in the matrix containing polyglycerol fatty acid ester and or lipid, the amount of viscous agent is about 0.005 to about 95% (weight), preferably about 0.5 to about 30% (weight), more Preferably about 1 to about 25% by weight, more preferably about 1 to about 20% by weight, based on total weight. When the substrate is coated with adhesive, the ratio of adhesive is also about 0.005 to about 95% by weight, preferably about 0.5 to about 30% by weight, more preferably about 1 to about 25% by weight, more preferably about 1 to about 20% by weight, based on the total weight.

When the composition of the present invention contains Alcaligenes polysaccharide as a swelling substance, the composition itself can adhere to the gastrointestinal mucosa even without adding said viscous agent because Alcaligenes polysaccharide itself functions as a viscous agent. In this case, the Alcaligenes polysaccharide is formulated in an amount exceeding the range defined above to impart the desired adhesive effect.

The gastric mucoadhesive composition comprising a viscous agent dispersed in a matrix containing polyglycerol fatty acid esters and/or lipids may be polyglycerol fatty acid esters and/or lipids, viscous agents, Alcaligenes polysaccharides and/or low Optional dispersion of substituted hydroxypropyl cellulose and active ingredient. Dispersion can be carried out analogously to known techniques.

The amount of compound (I) in the pharmaceutical composition of the present invention is generally 2 to 85% by weight, preferably 5 to 70% by weight.

The production methods of the pharmaceutical composition containing the compound (I) of the present invention include those known methods commonly used in the pharmaceutical field. In addition, the composition can be produced using an appropriate amount of excipients, binders, disintegrants, lubricants, sweeteners, surfactants, suspending agents, emulsifying agents and the like generally used in the pharmaceutical industry.

For producing tablets containing compound (I), for example, the excipients, binders, disintegrants and lubricants are used. For the production of pills or granules, with excipients, binders and disintegrants. Excipients are also used in the production of powders or capsules, while sweeteners are added in the production of syrups. In producing emulsions or suspensions, suspending agents, surfactants and/or emulsifying agents are added. Excipients include, but are not limited to, lactose, sucrose, dextrose, starch, sucrose, microcrystalline cellulose, hay powder, mannitol, sodium bicarbonate, calcium phosphate, and calcium sulfate. Binders include, but are not limited to, 5 to 10% by weight starch solutions, 10 to 20% by weight gum arabic solutions or gelatin solutions, 1 to 5% by weight tragacanth solutions, carboxymethyl cellulose solutions, Sodium alginate solution and glycerol. Disintegrants include, but are not limited to, starch and calcium carbonate. Lubricants include, but are not limited to, magnesium stearate, stearic acid, calcium stearate, and purified talc. Sweeteners include, but are not limited to, glucose, fructose, invert sugar, sorbitol, xylitol, glycerin, and simple syrup. Surfactants include, but are not limited to, sodium laurate sulfate, polysorbate-80, sorbitan fatty acid monoester and polyoxystearate-40. Suspending agents include, but are not limited to, acacia, sodium alginate, sodium carboxymethylcellulose, methylcellulose and bentonite. Emulsifiers include, but are not limited to, acacia, tragacanth, gelatin, and polysorbate-80. In addition, coloring agents, preservatives, fragrances, correctives, stabilizers, thickeners and other commonly used additives for pharmaceutical applications can also be added in appropriate amounts during the production of the dosage form containing compound (I) .

An example of a method for producing the adhesive composition for gastric mucosal diseases of the present invention will be described below.

1. The gastric mucoadhesive composition, which is solid at atmospheric temperature, can be produced in a manner analogous to known techniques. A typical method comprises melting polyglycerol fatty acid esters and/or lipids at a temperature exceeding their melting point, adding said viscous agent, anti-bacillus agent, and Alcaligenes polysaccharide and/or low Substituted hydroxypropyl cellulose in order to disperse them in the melt, and to cool the dispersion. The heating temperature may be, for example, about 40°C to about 150°C, preferably about 50°C to about 110°C, more preferably about 50°C to about 100°C. This process is carried out with a conventional granulator, and the composition is preferably molded into solid spheres (eg, granules, granules, etc.) by spray cooling, such as spray condensation.

The spray condensation method generally comprises dropping viscous agent, Alcaligenes polysaccharide and/or low-substituted Mixed dispersion of hydroxypropyl cellulose and active ingredient in molten polyglycerol fatty acid esters and/or lipids. The rotating disk may be, for example, a smooth circular disk, typically made of aluminum and measuring from about 5 to about 100 centimeters in diameter, preferably from about 10 to about 20 centimeters in diameter. The drop rate of the molten dispersion is selected according to the specified particle size, generally from about 1 to about 1000 g/min, preferably from about 2 to about 200 g/min, more preferably from about 5 to about 100 g/min. The granules thus obtained are indeed spherical, so that a homogeneous film can be formed on their surface with good efficiency in the subsequent coating step.

An alternative production method involves kneading the viscous agent, Alcaligenes polysaccharide and/or low-substituted hydroxypropylcellulose and the active ingredient into polyglycerol fatty acid ester and/or lipid and granulating the resulting dispersion. The solvent used in this method may be a common type of solvent (for example, methanol, acetonitrile, chloroform, etc.).

Another alternative method of producing solid compositions involves the use of melt granulation techniques. A typical melt granulation process involves heating the polyglycerol fatty acid ester and/or lipid at a temperature close to its melting point, e.g. from its melting point to a temperature about 5° C. below the melting point, and granulating the resulting melt, e.g., spray condensation as described above, And heating and suspending the obtained fine particles together with a viscous agent, an anti-bacillus agent, and Alcaligenes polysaccharide and/or low-substituted hydroxypropyl cellulose at a suitable temperature to provide a viscous matrix-drug system. In this case, disturbance of the active ingredients by heat can be avoided.

Compositions containing a matrix made of polyglycerol fatty acid esters and/or lipids and coated with viscous agents may be coated with such viscous agents alone or viscous agents and swelling substances (for example, Alcaligenes polysaccharide and/or low Substituted hydroxypropyl cellulose, etc.), the preferred coating material only contains viscous agent or viscous agent plus Alcaligenes polysaccharide and/or low-substituted hydroxypropyl cellulose. The coating material may be a composition containing at least one component selected from the polyglycerin fatty acid ester, the lipid, and the water-insoluble polymer. When a viscous agent that is less or incompatible with the components of the composition is used for the coating, a solid composition is provided that has a film in which the viscous agent is dispersed. The coating material may further contain the aforementioned additives.

Water-insoluble (hydrophobic) polymers include, but are not limited to, hydroxypropylmethylcellulose phthalate (Japanese Pharmacopoeia Twelfth Edition), hydroxypropylmethylcellulose acetate succinate (ShinEtsu Chemicals Co., Ltd.), carboxy Methyl ethyl cellulose (Freund Industries Co., Ltd., CMEC, Japanese Standards for Pharmaceutical Ingredients, 1986), cellulose acetate trimellitate (Eastman), cellulose acetate phthalate (Japanese Pharmacopoeia Twelfth Edition), Ethyl cellulose (Asahi Chemical Industry Co., Ltd.), aminoalkyl methacrylate copolymer (Röhm-Pharma, Eudragit ^TM RS-100, RL-100, RL-PO, RS-PO, RS-30D, RL -30D), methacrylic acid-ethyl acrylate copolymer (Rhm-Pharma, Eudragit ^TM L100-55, methacrylic acid-methyl methacrylate copolymer (Rhm-pharma, Eudragit ^TM L-100, S-100), Eudragit ^TM L30D-55, Eudragit ^TM NE-30D (Rhm-Pharma) and polyvinyl acetate (Colorcon). These hydrophobic polymers can be used alone or as a mixture of two or more different polymers use.

The ratio of viscous agent in the coating material is based on the solid fraction of the whole coating material, and is about 0.005 to about 100% (weight), preferably about 0.05 to about 95% (weight), more preferably about 0.05 to about 30 % by weight, more preferably about 1 to about 10% by weight.

When at least one polyglycerol fatty acid ester, lipid and hydrophobic polymer are used in combination with the viscous agent for the coating material, the proportion of the viscous agent based on the total weight of the solid fraction of the coating material is about 0.05 to about 95% (weight ), preferably about 0.5 to about 95% (weight), more preferably about 0.5 to 30% (weight), especially 5 to about 30% (weight), more preferably about 5 to about 25% (weight).

Further for the coating material, two or more components selected from polyglycerin fatty acid esters, lipids and hydrophobic polymers may be used in combination. In this case, based on each part by weight of all polyglycerol fatty acid esters and/or lipids, the remaining components are present in an amount of about 0.0001 to about 1000 parts by weight, preferably about 0.01 to about 100 parts by weight, more preferably about 0.01 to about 10 parts by weight. A ratio of parts by weight is used.

The amount of the coating material can be selected according to the kind of solid composition and the required adhesion strength to the gastric mucosa. For example, for tablets, the amount of the solid coating material may be about 0.1 to about 30% by weight, preferably about 0.5 to about 20% by weight; and for fine particles, about 0.1 to about 30% by weight; 100% by weight, preferably from about 1 to about 50% by weight.

If desired, the coating material may be supplemented with common additives, such as the aforementioned additives. For example, coating materials and additives can be applied together or added separately, etc. The proportion of the additive is about 0.1 to about 70% by weight, preferably about 1 to about 50% by weight, more preferably about 20 to about 50% by weight relative to the solid component of the coating material.

Applicable coating techniques include various well-known methods, such as pan coating, fluidized bed coating, roller coating, and the like. When the coating material is a solution or dispersion containing water or an organic solvent, spray coating can also be utilized. There is no particular limitation on the kind of the water or organic solvent. For example, alcohols such as methanol, ethanol, isopropanol and the like; ketones such as acetone; and halogenated hydrocarbons such as chloroform, dichloromethane, chloroform and the like can be used.

When polyglycerol fatty acid ester and/or lipid are used for coating, by melting polyglycerol fatty acid ester and/or lipid optionally together with other additives under heating, emulsifying the melt with water, and using the obtained emulsion The surface of the solid composition is sprayed and the coating is dried to produce the desired coating material composition. An alternative procedure involves adding the coating material to a preheated solid composition in a coating material pan or the like, and melting the spreading coating material.

The solid composition is generally coated at a temperature of from about 25°C to about 60°C, preferably from about 25°C to about 40°C.

The coating time can be reasonably selected with reference to the coating method, the properties and quantity of the coating material, and the properties of the solid composition of the substrate.

In order to ensure adequate adhesion to the gastrointestinal mucosa, if necessary, the gastric mucosa-adhesive solid composition can be further coated with commonly used gastric coating materials or water-soluble coating materials.

The gastric mucoadhesive composition of the present invention is generally orally administered as it is, or in the form of a suitable preparation. Solid oral dosage forms include, but are not limited to, fine granules, granules, pills, tablets made by compressing said fine granules or granules with a tablet machine and filling said fine granules or granules into suitable capsule shells. capsule. Among these formulations, fine granules and granules are preferred.

For example, the fine particles may have a particle size distribution of not less than about 75% by weight of particles measuring from about 10 to about 500 microns in diameter, not more than about 5% by weight of particles greater than about 500 microns, and not greater than about 10% by weight of particles smaller than about 10 microns. A preferred distribution is about > 75% by weight from about 105 to about 500 microns, not more than about 5% by weight from about > 500 microns, and not more than about 10% by weight from about ≤ 74 microns. The particle size distribution of the granules can be, for example, not less than about 90% by weight from about 500 to about 1410 microns, and not more than about 5% by weight from about ≤ 177 microns.

2. When the gastric mucoadhesive composition is provided as a liquid composition, this liquid composition can be produced in a manner similar to known techniques. Typical methods include making polyglycerol fatty acid esters and/or lipids (liquid at atmospheric temperature), viscosifiers, active ingredients and swelling substances (e.g., Alcaligenes polysaccharide and/or low-substituted hydroxypropyl cellulose, etc. ) are mixed all at once or sequentially to provide a dispersion or solution.

Dosage forms containing such liquid mucosa-adhesive pharmaceutical systems include, but are not limited to, syrups, emulsions, suspensions, and capsule forms thereof.

The proportion of active ingredients (for example, anti-H. pylori agent etc.) in the composition of the present invention is about 0.005 to about 95% (weight), preferably about 1 to about 95% (weight), more preferably about 10 to about 95% % (weight), more preferably about 10 to about 50.

The pharmaceutical composition (especially the gastric mucosal adhesive composition) of the present invention comprising compound (I) or its salt is stable and has little toxicity, so it can be used safely. The daily dosage varies according to the clinical condition and body weight of the patient, the specific species of the compound and the route of administration; 500 mg, preferably about 10 to 200 mg of the active ingredient (compound (I) or a salt thereof).

In the pharmaceutical composition of the present invention, compound (I) can be used in combination with one or more other antibacterial and/or antiulcer agents.

Therefore, other antimicrobial agents that may be included include, but are not limited to, nitroimidazoles (eg, tinidazole and metronidazole), tetracyclines (eg, tetracycline, doxycycline, and minocycline), penicillins (eg, amoxicillin, ampicillin, and mezlocillin), cephalosporins (eg, cefaclor, cefadroxil, cefazolin, cefuroxime, cefuroxime axetil, cephalexin, cefpodoxime axetil, ceftazidime and ceftriaxone), carbapenems (e.g., imipenem and meropinine), aminoglycosides (e.g., paromomycin), macrolides (e.g., erythromycin , erythromycin and azithromycin), lincosamides (eg, clindamycin), rifamycins (eg, rifampicin) and nitronitrofurantoin. As antiulcer drugs used in combination with compound (I), gastric proton pump inhibitors (for example, lansoprazole and omeprazole, pantoprazole, rabeprazole, lemirazole) and _H2 receptor antagonists (eg, ranitidine, cimetidine, and famotidine).

The other antibacterial agents and/or antiulcer agents mentioned above may be used in combination of two or more. In this combination drug therapy, the daily dose (for adults) of said other antibacterial agents or drugs is 1 to 500 mg, preferably 5 to 200 mg, and the daily dose (for adults) of said other antiulcer drugs is 0.5 to 500 mg. 1000 mg, preferably 1 to 500 mg.

Best Mode for Carrying Out the Invention

The following examples, test examples and preparation examples are only used to illustrate the present invention more specifically, but not to limit the scope of the present invention. In these examples, percentages (%) mean weight/volume percentages unless otherwise stated. The mixing ratio of solvents is a volume ratio unless otherwise indicated. NMR spectra were recorded using a Bruker AC-300 spectrometer or a Varian gemini 200 spectrometer.

Example 1

(S)-3-[(2S,3R,4R,5S)-2,3,4,6-tetrahydroxy-5-(L-valyl-L-leucyl)aminocaproyl]amino-3 -Phenylpropanoic acid (HC-70II, compound 2) and (S)-3-[(2S,3R,4R,5S)-2,3,4,6-tetrahydroxy-5-(L-leucyl )aminocaproyl]amino-3-phenylpropionic acid (HC-70III, compound 3)

On the slant medium that is made up of glucose 0.1%, tryptone 0.5%, yeast extract 0.25%, and agar 1.5%, the bacterium ring bacillus HC-70 that fully grows is used for inoculating and is equipped with 500 milliliters of seed culture medium (pH =7.0) of 2 liters of Sakaguchi flask, the culture medium consists of glucose 2.0%, soluble starch 3.0%, corn steep liquor 0.3%, soybean flour 1.0%, polypeptone 0.5%, yeast extract 0.1%, oatmeal agar 0.2%, Sodium chloride 0.3% and precipitated calcium carbonate 0.5% constituted and incubation was carried out at 24°C for 2 days on a reciprocating shaker. 500 ml of culture was transferred to a 200 liter fermenter with 120 liters of production medium (pH=6.5) consisting of glucose 0.5%, dextrin 5.0%, soybean flour 3.5%, yeast extract 0.5%, sediment Calcium carbonate 0.7%, ACTOCOL ^TM 31-56 (Takeda Chemical Industry Co., Ltd.) 0.05% and silicone oil 0.05%, and the fermentation was carried out at a temperature of 22° C. and an internal pressure of 1.0 kg/cm 2 under aeration of 120 liters/minute and 120 rpm This was carried out for 42 hours with stirring.

The resulting culture solution (120 liters) was adjusted to pH = 7 and filtered with a filter aid (Radiolite 600, Showa Chemical Industry Co., Ltd.). The filtrate (130 L) was adjusted to pH = 7 and purified by HP-20 (7 L) column chromatography. After washing the column with water (21 L), elution was performed with 30% (v/v) isopropanol/water (28 L). The eluate was concentrated, and the residue was diluted with water to a volume of 30 liters and purified by CNP-80 (H-type, 15 liters) column chromatography. After washing the column with water (45 L), it was eluted with 2N-ammonia (53 L). The eluate was concentrated and purified by PA-412 (OH-form, 2 L) column chromatography. The column was washed successively with water (6 L) and 1M NaCl/water (2 L) and eluted successively with 1M NaCl/water (10 L) and 1N hydrochloric acid (4 L). The eluate was adjusted to pH 7, and purified by HP-20 (1 L) column chromatography. The column was washed with water (3 L) and eluted with 30% (v/v) isopropanol/water (3.4 L). The eluate was concentrated, adjusted to pH 7, and purified by HP-20S (400 mL) column chromatography. After washing the column with water (1.2 L), it was eluted sequentially with 5% (v/v) isopropanol/water (1.2 L) and 10% (v/v) isopropanol/water (1.2 L). The 5% (v/v) isopropanol/water eluate was concentrated and purified by HP-20SS (100 mL) column chromatography. Wash the column with water (200 ml) and wash with water (100 ml), 2% (v/v) isopropanol/water (300 ml) and 5% (v/v) isopropanol/water (300 ml) in sequence elute. The eluate was concentrated and allowed to stand at 7°C, and the crystals were collected to give HC-70III (compound 3; 1.3 g). The 10% (v/v) isopropanol/water eluate from the HP-20S (400 mL) column was concentrated, after addition of methanol, the concentrate was allowed to stand at 7°C and the resulting crystals (1.7 g) were collected by filtration. This crystal was recrystallized twice from water. In this way, crystals (1.3 g) mainly composed of HC-70II were obtained. Of these crystals, 719 mg was subjected to HP-20S (70 ml) column chromatography. The column was washed with water (210 ml), 2% (v/v) isopropanol/water (210 ml) and 5% (v/v) isopropanol/water (210 ml), and washed with 10% (v/v) ) isopropanol/water (420 mL) for elution. The HC-70II fraction was concentrated and allowed to stand at 7°C, and the resulting crystals were recovered by filtration to give HC-70II (compound 2; 479 mg).

Example 2

(S)-3-[(2S,3R,4R,5S)-2,3,4,6-tetrahydroxy-5-(L-valyl-L-valyl-L-leucyl)amino Hexanoyl]amino-3-phenylpropanoic acid (HC-70I, compound 1)

On the slant medium that is made up of glucose 0.1%, tryptone 0.5%, yeast extract 0.25%, and agar 1.5%, the bacterium ring bacillus HC-70 that fully grows is used for inoculating and is equipped with 500 milliliters of seed culture medium (pH =7.0) of 2 liters of sterile Sakaguchi flasks, the culture medium consists of glucose 2.0%, soluble starch 3.0%, corn steep liquor 0.3%, soybean flour 1.0%, polypeptone 0.5%, yeast extract 0.1%, oatmeal agar 0.2% %, sodium chloride 0.3% and precipitated calcium carbonate 0.5%, and the incubation was carried out on a reciprocating shaker at 24°C for 2 days. 500 ml of culture was transferred to a 200 liter fermenter with 120 liters of production medium (pH=6.5) consisting of glucose 0.5%, dextrin 5.0%, soybean flour 3.5%, yeast extract 0.5%, sediment Calcium carbonate 0.7%, ACTOCOL ^TM 31-56 (Takeda Chemical Industry Co., Ltd.) 0.05% and silicone oil 0.05%, and the fermentation was carried out at a temperature of 22° C. and an internal pressure of 1.0 kg/cm 2 under aeration of 120 liters/minute and 120 rpm Stirring was carried out for 24 hours.

Two equivalent batches of the resulting broth (120 liters) were filtered with a filter aid (Radiolite 600). The filtrate (245 L) was adjusted to pH=6 and purified by HP-20 (15 L) column chromatography. After washing the column with water (45 L), elution was performed with 30% (v/v) isopropanol/water (60 L). The eluate was purified by CNP-80 (H-type, 20 liters) column chromatography, washed with water (60 liters) and then eluted with 2N-ammonia water (80 liters). The eluate was concentrated, adjusted to pH=6, and purified by HP-20 (2.4 L) column chromatography. After washing the column with water (7.2 liters) and 5% (volume/volume) isopropanol/water (7.2 liters) successively, wash the column with 10% (volume/volume) /vol) isopropanol/water (11.5 L) for elution. The eluate was concentrated and passed through IR-120 (sodium-form, 1.5 L), IRA-67 (hydroxyl-form, 1.5 L) and SP-850 (2 L) columns successively. After washing with water (8 L), the SP-850 (2 L) was further washed with 0.2N ammonia (2 L), water (6 L) and 10% (v/v) isopropanol/water (2 L) in the above order. ) column. Then, it was eluted with 10% (v/v) isopropanol/water (4 L). The eluate was concentrated and allowed to stand at 7°C, and the resulting crystals (1.4 g) were harvested by filtration. Of these crystals, 1.2 g was subjected to HP-20 (150 ml) column chromatography. After the chromatographic column was washed successively with water (450 ml) and 2% (volume/volume) isopropanol/water (450 ml), it was washed with 5% (volume/volume) isopropanol/water (900 ml), 10% (volume /vol) isopropanol/water (900 mL) and 15% (vol/vol) isopropanol/water (450 mL) were eluted sequentially. The HC-70I fraction was concentrated and allowed to stand at 7°C, and the resulting crystals were recovered to obtain HC-70I (compound 1; 199 mg).

Example 3

(S)-3-[(2S,3R,4R,5S)-2,3,4,6-tetrahydroxy-5-(L-valyl-L-leucyl)aminocaproyl]amino-3 -Phenylpropionate hydrochloride (HC-70II monohydrochloride, compound 4)

To HC-70II (compound 2; 200 mg) were added 0.1N hydrochloric acid (3.4 ml) and water (60 ml), and the mixture was warmed to prepare a solution. This solution was filtered through DISMIC-25CS (0.45 µm, Toyo Roshi) and the filtrate was lyophilized to give HC-70II monohydrochloride (compound 4; 199 mg).

Elemental analysis: (C ₂₆ H ₄₂ N ₄ O ₉ ·HCl·2.5H ₂ O)

Found: C, 48.93; H, 7.18; N, 8.86; Cl, 5.64

Analytical values: C, 49.09; H, 7.61; N, 8.81; Cl, 5.57

Example 4

(S)-3-[(2S,3R,4R,5S)-5-amino-2,3,4,6-tetrahydroxyhexanoyl]amino-3-phenylpropanoic acid (compound 5)

HC-70III (compound 3; 190 mg) was dissolved in phosphate buffer (40 mmol, pH 8; 47.5 mL), then cobalt chloride (1 mol, 0.19 mL) and Actinase E (19 mg, Kaken Pharmaceutical Co. ) in aqueous solution and reacted at 37°C for 2 hours. The reaction mixture was filtered through filter paper (No. 2, Toyo Roshi), and the filtrate was subjected to HP-20 (50 ml) column chromatography for purification. The column was washed with water (50 mL) and eluted sequentially with water (100 mL) and 20% (vol/vol) isopropanol/water (200 mL). The eluate was concentrated and lyophilized to give a crude powder (149 mg).

The above coarse powder is carried out to preparative HPLC [column: YMC-Pack SH-363-15, ODS (YMC), mobile phase: 5% (volume/volume) acetonitrile/0.02 mole phosphate buffer (pH4.5), flow velocity: 12 ml/min]. Fractions 400-600 mL were pooled, pH adjusted to 7 and concentrated under reduced pressure to 120 mL. The concentrate was chromatographed on a HP-20 (60 mL) column and, after washing with water (180 mL), was eluted with 20% (vol/vol) isopropanol/water (240 mL). The eluate was concentrated and lyophilized to afford compound 5 as a white powder (103 mg).

¹³ C-NMR (DMSO-d ₆ , δppm): 174.9, 172.3, 143.4, 127.9,

126.3, 126.2, 71.4, 70.8, 66.6, 60.9, 53.3, 49.7, 43.1

Elemental analysis: (C ₁₅ H ₂₂ N ₂ O ₇ 1.5H ₂ O)

Found values: C, 49.11; H, 6.78; N, 7.89

Analytical values: C, 48.78; H, 6.82; N, 7.58

Example 5

(Use Bacillus aneurysm HC-72 to obtain HC-70III)

By glucose 0.1%, tryptone 0.5%, yeast extract 0.25%, and the slant medium that agar 1.5% constitutes fully grows a bacterium loop bacillus aneurysm HC-72 and is used for inoculating and is equipped with 500 milliliters of seed culture medium ( pH=7.0) in a 2 liter Sakaguchi flask, the medium was composed of glucose 2.0%, soluble starch 3.0%, corn steep liquor 0.3%, soybean flour 1.0%, polypeptone 0.5%, yeast extract 0.1%, sodium chloride 0.3 % and precipitated calcium carbonate 0.5%, and the incubation was carried out on a reciprocating shaker at 28°C for 1 day. 500 milliliters of cultures were transferred to 200 liters of fermenters with 120 liters of production medium (pH=7.0). The production medium consisted of 2.0% glucose, 3.0% soluble starch, 0.3% corn steep liquor, 1.0% soybean flour, poly Peptone 0.5%, yeast extract 0.1%, sodium chloride 0.3%, precipitated calcium carbonate 0.5%, ACTOCOL ^TM 31-56 (Takeda Chemical Industry Co., Ltd.) 0.05% and silicone oil 0.05%, and the incubation was at a temperature of 24°C It was carried out for 48 hours with an internal pressure of 1.0 kg/cm2, aeration at 120 liters/minute and stirring at 120 rpm. 10 liters of culture were transferred to 2000 liters of fermenters equipped with 1200 liters of production medium (pH 7.0) consisting of 0.5% glucose, 1.0% inositol, 5.0% soybean flour, 1.0% corn steep liquor, ACTOCOL ^TM 31-56 (Takeda Chemical Industry Co., Ltd.) 0.05% and silicone oil 0.05%, and the incubation was carried out for 114 hours at a temperature of 28° C. and an internal pressure of 1.0 kg/cm 2 , aeration at 840 liters/minute, and stirring at 30 rpm.

The obtained fermentation broth (1200 liters) was adjusted to pH 5, and flocculant [0.5 (weight/volume) Sanfloc C-109P, Sanyo Chemical Industry Co., Ltd.] was added to carry out flocculation. Then, the fermentation broth was filtered with a filter aid (Radiolite 600). The filtrate (1200 L) was adjusted to pH 5 and purified by activated carbon (Granular Shirasagi, 25 L) and SP-850 (100 L) column chromatography, and washed with water (300 L). A single SP-850 column was washed sequentially with 0.1N sodium hydroxide/water (300 liters), water (300 liters), 0.1N sulfuric acid (300 liters) and water (300 liters), and washed with 25% (v/v) iso Propanol/water (400 L) was used for elution. The HC-70III fraction was adjusted to pH 4.5, and purified by UBK-510L (sodium-form, 150 L) column chromatography. After washing the chromatographic column with water (150 liters), 0.01N-ammonia water (600 liters) was used for fractional elution. The HC-70III fraction was adjusted to pH 8.0 and columned through PK-216 (sodium-form, 25 liters) and IRA-67 ( _CH3COO -form, 25 liters) in the order described, followed by water (100 liters )washing. The effluent and washings were combined, adjusted to pH 5, concentrated, and allowed to stand at 7°C. The crystals were harvested by filtration to afford HC-70III (compound 3; 380 g).

Example 6

(S)-3-[(2S,3R,4R,5S)-2,3,4,6-tetrahydroxy-5-(L-valyl-L-isoleucyl)aminocaproyl-L- Leucyl]amino-3-phenylpropanoic acid (HC-70I-A, compound 1A) and (S)-3-[(2S,3R,4R,5S)-2,3,4,6-tetrahydroxy -5-(L-valyl-L-leucyl-L-leucyl)aminocaproyl]amino-3-phenylpropionic acid (HC-70I-B, compound 1B)

On the slant medium that is made up of glucose 0.1%, tryptone 0.5%, yeast extract 0.25%, and agar 1.5%, the bacterium ring bacillus HC-70 that fully grows is used for inoculating and is equipped with 500 milliliters of seed culture medium (pH = 6.5) in a 2 liter Sakaguchi flask, the medium was composed of glucose 2.0%, soluble starch 3.0%, corn steep liquor 0.3%, soybean flour 1.0%, polypeptone 0.5%, yeast extract 0.1%, sodium chloride 0.3% and precipitated calcium carbonate 0.5%, and the incubation was carried out on a reciprocating shaker at 24°C for 2 days. 500 ml of culture was transferred to a 200 liter fermenter with 120 liters of production medium (pH=6.5) consisting of dextrin 5.0%, glucose 0.5%, soybean flour 3.5%, yeast extract 0.5%, sediment Calcium carbonate 0.5%, ACTOCOL ^TM 31-56 (Takeda Chemical Industry Co., Ltd.) 0.05% and silicone oil 0.05%, and the fermentation was carried out at a temperature of 22° C. and an internal pressure of 1.0 kg/cm2, aeration at 120 liters/minute, and stirring at 120 rpm for 42 hours.

Two equivalent batches of the resulting broth (120 liters) were filtered with a filter aid (Radiolite 600). The filtrate (250 L) was adjusted to pH = 6 and purified by HP-20 (15 L) column chromatography. After washing the column with water (45 L), elution was performed with 30% (v/v) isopropanol/water (60 L). The eluate was purified by CNP-80 (H-type, 20 L) column chromatography, and after washing the column with water (60 L), it was eluted with 2N-ammonia water (80 L). The eluate was concentrated, adjusted to pH 6, and column-passed through IR-120 (NH ₄ -type, 1.5 L), IRA-67 (OH-type, 1.5 L) and SP-850 (2 L) in the order described, It was then washed with water (8 L). SP-850 (2 liters) column was sequentially filled with 0.2N ammonia (2 liters), water (6 liters), 0.1N hydrochloric acid (2 liters), water (6 liters), and 5% (v/v) isopropanol /water (6 L) in the above sequence and eluted fractionally with 20% (v/v) isopropanol/water (8 L) and 30% (v/v) isopropanol/water (6 L). Then, the fraction (11.5 liters) containing HC-70I-A and HC-70I-B was passed through IR-120 (NH ₄ -type, 1.5 liters) and IRA-67 (CH ₃ COO-type , 0.5 L). The effluent was concentrated and allowed to stand at 7°C, and the resulting crystals (9.6 g) were recovered by filtration. In this crystal, 4.0 g was dissolved in N,N-dimethylformamide and subjected to preparative HPLC [instrument: LC-300G, column: 100Φ×1000L (mm) (KuritaWater Industries Ltd.), stationary phase: YMC. GEL KE-ODS-10S (YMC), mobile phase: 17% (v/v) acetonitrile/0.02M phosphate buffer (pH4.5), flow rate: 30 ml/min] to obtain HC-70I-A fraction (60-85 minutes) and HC-70I-B fraction (87-100 minutes). The HC-70I-A fraction was concentrated and purified by HP-20 (100 mL) column chromatography. After washing the column with water (300 mL), fractionation was performed with 20% (v/v) isopropanol/water (600 mL) and 0.1N ammonia/20% (v/v) isopropanol/water (600 mL) elute. The HC-70I-A fraction (600 mL) was concentrated and allowed to stand at 7°C and the resulting crystals were collected by filtration to give HC-70I-A (compound 1A, 249 mg). The HC-70I-B fraction (0.4 L) eluted from the preparative HPLC column was concentrated and allowed to stand at 7°C, and the resulting crystals were recovered by filtration to give HC-70I-B (compound 1B; 400 mg).

HC-70I-A (compound 1A)

Optical rotation: -67° (c=0.50, 0.1N HCL, 21°C)

FAB-MS: m/z 668 (M+H) ⁺

Elemental analysis (%) (calculated based on 2 moles of water)

Found values: C, 54.54; H, 8.16; N, 10.12

Analysis value: C, 54.61; H, 8.16; N, 9.95 Molecular formula: C ₃₂ H ₅₃ N ₅ O ₁₀ ¹³ C-NMR spectrum (DMSO-d ₆ , 172.7, 172.4,

172.3, 172.2, 170.8, 142.6, 127.9, 126.5, 71.0, 70.8,

67.2, 60.5, 58.8, 56.5, 51.1, 50.8, 49.0, 41.2, 40.5,

36.7, 30.8, 24.3, 24.0, 23.1, 21.3, 19.0, 16.9, 15.3,

10.8 Amino acid analysis: analysis was performed after hydrolysis in 6N hydrochloric acid at 110° C. for 72 hours.

Leucine (1 mole), Isoleucine (1 mole), Valine (1 mole) High Performance Liquid Chromatography (HPLC)

Column: YMC-Pack ODS-A, A312, 150×6.0 mm (YMC)

Mobile phase: 15% (v/v) acetonitrile/0.02M phosphate buffer (PH4.5)

Flow rate: 1.0ml/min

Detection: Ultraviolet absorption method, 214 nm

Retention time: 27 minutes Thin layer chromatography (TLC):

Stationary phase: silica gel 60F ₂₅₄ , 0.25 mm (Merck, Germany)

Developing solvent: n-butanol/acetic acid/water (12:3:5)

Rf: 0.51HC-70I-B (Compound 1B) Optical rotation: -75° (c=0.50, 0.1N HCL, 21°C) FAB-MS: m/z 668 (M+H) ⁺ elemental analysis (%) ( Calculated based on 3.5 moles of water)

Found values: C, 52.59; H, 8.03; N, 9.78

Analytical values: C, 52.59; H, 8.27; N, 9.58 Molecular formula: C ₃₂ H ₅₃ N ₅ O ₁₀

¹³ C-NMR spectrum (in DMSO-d ₆ , δppm): 172.8, 172.5,

172.3, 171.9, 142.6, 128.0, 126.4, 71.0, 70.8, 67.2,

60.5, 59.0, 51.0, 50.7, 49.1, 41.2, 40.9, 40.4, 30.8,

24.1, 23.1, 22.9, 21.6, 21.3, 19.0, 16.9

Amino acid analysis: analysis after hydrolysis in 6N hydrochloric acid at 110°C for 72 hours

Leucine (2 moles), Valine (1 mole)

High Performance Liquid Chromatography (HPLC)

Column: YMC-Pack ODS-A, A312, 150×6.0 mm (YMC)

Mobile phase: 15% (v/v) acetonitrile/0.02M phosphate buffer (pH 4.5)

  Flow rate: 1.0ml/min

 Detection: UV absorption method, 214 nm

  Retention time: 39 minutes

Thin layer chromatography (TLC):

Stationary phase: silica gel 60F ₂₅₄ , 0.25 mm (Merck, Germany)

Developing agent: n-butanol/acetic acid/water (12:3:5)

Rf: 0.54

Example 7

(S)-3-[(2S,3R,4R,5S)-5-(N-acetyl-L-leucyl)amino-2,3,4,6-tetrahydroxyhexanoyl]amino-3-benzene Glypropionic acid monosodium salt (compound 6)

HC-70III (Compound 3; 50 mg) was dissolved in aqueous potassium bicarbonate (50 mmol, 20 mL), and then acetic anhydride (22 μL) was added to react at room temperature for 1 hour. The reaction mixture was adjusted to pH 6.5 and purified by HP-20 (5 mL) column chromatography. After washing the column with water (5 mL), it was eluted with water (10 mL) and 30% (v/v) isopropanol/water (30 mL). The eluate was concentrated and lyophilized to afford the title compound (compound 6; 49 mg).

¹ H-NMR (DMSO-d ₆ , δppm): 0.84 (3H, d, J=6.4Hz),

0.87(3H, d, J=6.5Hz), 1.44(2H, t, J=7.2Hz), 1.57(1H, m),

1.83(3H, s), 2.53(2H, d, J=6.8Hz), 3.45(2H, m),

3.48(1H, d, J=9.8Hz), 3.76(1H, d, J=9.8Hz), 3.95(1H, q

like), 4.12(1H, s), 4.30(1H, q like), 5.11(1H, q like),

7.16(1H, m), 7.23(2H, t like), 7.32(1H, d, J=7.0Hz),

7.33(2H, t like), 8.05(1H, d, J=8.3Hz),

8.75(1H, d, J=7.7Hz).

FAB-MS: m/z 536 (M+H) ⁺

Example 8

(S)-3-[(2S,3R,4R,5S)-2,3,4,6-tetrahydroxy-5-(L-leucyl)aminocaproyl]amino-3-phenylpropanoic acid Ester monohydrochloride (compound 7)

HC-70III (compound 3; 50 mg) was dissolved in methanol (10 ml), then hydrochloric acid-methanol reagent 10 (10 ml, Tokyo Kasei Kogyo) was added, and reacted at room temperature for 16 hours. Under nitrogen, the reaction mixture was concentrated to dryness, diluted with water (10 mL), adjusted to pH 6.5, further washed with water (40 mL), and purified by HP-20 (10 mL) column chromatography. The column was washed with water (30 ml) and 30% (v/v) isopropanol/water (10 ml), then washed with 30% (v/v) isopropanol/water (20 ml) and 50% (v/v) /vol) isopropanol/water (20 mL) for elution. The eluate was concentrated and lyophilized to afford the title compound (compound 7; 34 mg).

¹ H-NMR (DMSO-d ₆ , δppm): 0.86 (3H, d, J = 6.6 Hz),

0.89 (3H, d, J=6.7Hz), 1.24 (1H, ddd, J=4.8, 9.2, 13.5Hz),

1.46(1H, ddd, J=4.4, 9.0, 13.5Hz), 1.75(1H, m),

2.84(1H,dd,J=7.4,15.8Hz), 2.91(1H,dd,J=6.8,15.8Hz),

3.40-3.48(3H, m), 3.51(3H, s), 3.76(1H, m), 3.97(1H, q

like), 4.11 (1H, br d, J = 4.3Hz), 4.56 (1H, d, J = 6.3Hz),

4.63(1H, br s), 4.90(1H, br d, J=6.0Hz), 5.24(1H, br

d, J=7.0Hz), 5.27(1H, q like), 7.20-7.37(5H, m),

7.45(1H, br d, J=7.8Hz), 8.12(1H, br d, J=8.9Hz).

FAB-MS: m/z 470 (M+H) ⁺

Reference example 1

(2S,3R,4R,5S)-2,3,4,6-tetrahydroxy-5-(L-leucyl)aminocaproic acid (De-β_Phe-HC-70III)

HC-70III (compound 3; 910 mg) was dissolved in 0.5N sodium hydroxide/water (200 mL) and the solution was stirred at 37°C for 24 hours. The reaction mixture was adjusted to pH 5 and purified by SP-207 (100 mL) column chromatography. The chromatographic column was washed with water (300 ml), and the effluent and washings were combined and purified by activated carbon (Granular Shirasagi, 70 ml) column chromatography. After washing the column with water (210 mL), it was eluted with 10% (vol/vol) isopropanol/water (210 mL). The eluate was concentrated and column-passed through Sephadex G-10 (600 mL), eluted stepwise with water (600 mL). Fractions containing De-β-Phe-HC-70III were concentrated to dryness, and the residue was washed with water (2 ml) and ethanol (4 ml) and allowed to stand at 7°C. The resulting crystals were harvested by filtration to give the title compound (De-β-Phe-HC-70III; 300 mg).

¹ H-NMR (DMSO-d ₆ , δppm): 0.86 (3H, d, J=7.1 Hz),

0.89(3H, d, J=7.3Hz), 1.36(1H, m), 1.49(1H, m), 1.67(1H, m),

3.30(1H,dd,J=3.9,9.3Hz), 3.38(1H,dd,J=6.3,9.8Hz),

3.43(1H, dd, J=9.3, 9.8Hz), 3.58(1H, t like),

3.70(1H,d,J=9.3Hz), 3.85(1H,d,J=3.9Hz), 4.05(1H,d

like), 7.90 (1H, d, J=8.6Hz).

FAB-MS: m/z 309 (M+H) ⁺

Reference example 2

(2S,3R,4R,5S)-5-(N-Benzyloxycarbonyl-L-leucyl)amino-2,3,4,6-tetrahydroxyhexanoic acid diphenylmethyl ester

To (2S,3R,4R,5S)-2,3,4,6-tetrahydroxy-5-(L-leucyl)aminocaproic acid (300mg) in water (20ml) and tetrahydrofuran (5ml) To the solution were added benzyl chloroformate (0.167 ml) and sodium bicarbonate (245 mg), and the mixture was stirred at room temperature for 3 hours. After distilling off THF, the aqueous layer was acidified with 1N hydrochloric acid (3 mL) and extracted with ethyl acetate (100 mL×2). The extract was washed with saturated brine and dried over anhydrous sodium sulfate. After concentration under reduced pressure, the residue was dissolved in methanol (10 ml), and diphenyldiazomethane (400 mg) was added. The whole was stirred at room temperature for 14 hours and concentrated under reduced pressure. The residue was purified by silica gel column chromatography, and then eluted with ethyl acetate-methanol (10:1). The effective fractions were combined and concentrated under reduced pressure to give the title compound (495 mg).

¹ H-NMR (CD ₃ OD) δ: 0.86-0.90 (6H, m), 1.49-1.76 (3H, m),

3.62-3.69(2H,m), 3.83(1H,m), 3.99(1H,m), 4.16-

4.27(2H, m), 4.35(1H, m), 4.56(1H, d, J=1.8Hz), 5.05-

5.08(2H, m), 6.91(1H, s), 7.24-7.38(15H, m).

Reference example 3

(2S,3R,4R,5S)-2,3,4,6-tetraacetoxy-5-(N-benzyloxycarbonyl-L-leucyl)aminocaproic acid diphenylmethyl ester

To (2S, 3R, 4R, 5S)-5-(N-benzyloxycarbonyl-L-leucyl)amino-2,3,4,6-tetrahydroxyhexanoic acid diphenylmethyl ester (200 mg) To a solution of pyridine (5 ml) were added acetic anhydride (3 ml) and dimethylaminopyridine (40 mg) and the mixture was stirred at room temperature for 16 hr. After concentration under reduced pressure, 1N hydrochloric acid (10 mL) was added to the concentrate and the whole was extracted with ethyl acetate (100 mL×2). The extract was washed with saturated brine and dried over anhydrous sodium sulfate. Removal of the organic solvent gave a residue which was chromatographed on a silica gel column and eluted with ethyl acetate-hexane (1:2). The effective fractions were combined and concentrated under reduced pressure to give the title compound (256 mg).

¹ H-NMR (CDCl ₃ ) δ: 0.91-0.95 (6H, m), 1.46-1.69 (3H, m),

1.81(3H,s), 1.98(3H,s), 2.07(3H,s), 2.17(3H,s),

3.86(1H, dd, J=11.4Hz, 6.6Hz), 3.99-4.17(2H, m),

4.47(1H, m), 5.01-5.09(2H, m), 5.22(1H, d, J=1.8Hz),

5.40(1H, d, J=9.6Hz), 5.52(1H, d, J=9.6Hz),

6.37(1H, d, J=8.8Hz), 6.76(1H, s), 7.26-7.34(16H, m).

Reference example 4

(2S,3R,4R,5S)-2,3,4,6-Tetraacetoxy-5-(N-benzyloxycarbonyl-L-leucyl)aminocaproic acid

Diphenylmethyl (2S,3R,4R,5S)-2,3,4,6-tetraacetoxy-5-(N-benzyloxycarbonyl-L-leucyl)aminocaproate (270 mg) It was dissolved in trifluoroacetic acid (5 ml), and the solution was stirred at room temperature for 1 hour. After concentration under reduced pressure, the residue was purified by silica gel column chromatography, and then eluted with ethyl acetate-methanol (10:1). Effective fractions were combined and concentrated under reduced pressure to give the title compound (214 mg).

¹ H-NMR (CD ₃ OD) δ: 0.92-0.96 (6H, m), 1.50-1.67 (3H, m),

1.96(3H,s), 2.04(3H,s), 2.06(3H,s), 2.11(3H,s),

3.83(1H, m), 4.11-4.23(3H, m), 4.52(1H, m), 4.95(1H, m),

5.07(1H, m), 5.36(1H, d, J=10.0Hz), 5.46(1H, d, J=10.0Hz),

7.28-7.32(5H, m).

Example 9

(S)-3-[(2S,3R,4R,5S)-2,3,4,6-tetrahydroxy-5-(L-leucyl)aminocaproyl]amino-3-phenylpropionate salt

To (S)-3-[(2S,3R,4R,5S)-2,3,4,6-tetrahydroxy-5-(L-leucyl)aminocaproyl]amino-3-phenylpropanoic acid To a solution of (Compound 3) (2.00 g) in water (50 ml) was added 1N hydrochloric acid (4.83 ml). The mixture was filtered through a membrane filter, and the filtrate was concentrated under reduced pressure. Recrystallization from methanol-ether gave the title compound (1.84 g).

¹ H-NMR (CD ₃ OD) δ: 1.02 (3H, d, J = 6.4 Hz),

1.03(3H, d, J=6.4Hz), 1.60-1.80(3H, m),

2.85(1H, dd, J=16.0Hz, 7.0Hz), 3.65-4.40(7H, m), 5.30-

5.50(1H, m), 7.20-7.45(5H, m), 8.42(1H, d, J=8.6Hz).

Example 10

(S)-3-[(2S,3R,4R,5S)-2,3,4,6-tetrahydroxy-5-(L-leucyl)aminocaproyl]amino-3-phenylpropionic acid di Phenyl methyl ester

To (S)-3-[(2S,3R,4R,5S)-2,3,4,6-tetrahydroxy-5-(L-leucyl)aminocaproyl]amino-3-phenylpropanoic acid To a solution of hydrochloride (1.84 g) in methanol (40 ml) was added diphenyldiazomethane (1.45 g) in methanol (20 ml), and the mixture was stirred at room temperature for 4 hours. After adding acetic acid (0.1 mL), the reaction mixture was extracted with ethyl acetate. The extract was washed with aqueous sodium bicarbonate solution and saturated brine, respectively, and the ethyl acetate solution was dried over anhydrous sodium sulfate. Removal of the organic solvent gave a residue, which was purified by silica gel column chromatography. Elution with ethyl acetate-methanol (2:1) gave the title compound (2.05 g).

¹ H-NMR (CD ₃ OD) δ: 0.93 (3H, d, J = 5.0 Hz),

0.96(3H, d, J=4.8Hz), 1.10-1.90(3H, m),

3.02(1H,dd,J=15.8Hz,7.6Hz), 3.13(1H,dd,J=15.8Hz,5.8Hz),

3.35-4.35(7H, m), 5.35-5.50(1H, m), 6.73(1H, s), 7.10-

7.40(15H, m).

Example 11

(S)-3-[(2S,3R,4R,5S)-2,3,4,6-tetrahydroxy-5-(L-leucyl)aminocaproyl]amino-3-phenylpropionic acid di Phenyl methyl ester hydrochloride

At room temperature, to (S)-3-[(2S,3R,4R,5S)-2,3,4,6-tetrahydroxy-5-(L-leucyl)aminocaproyl]amino-3-benzene To a solution of diphenylmethyl phenylpropionate (246 mg) in methanol (1 ml) was added 1N hydrochloric acid (0.396 ml). Concentration gave a residue, which was recrystallized from methanol-ether to give the title compound (180 mg).

¹ H-NMR (CD ₃ OD) δ: 1.00 (3H, d, J = 5.6 Hz),

1.02(3H, d, J=5.6Hz), 1.60-1.80(3H, m),

3.04(1H,dd,J=16.0Hz,7.0Hz), 3.15(1H,dd,J=16.0Hz,5.4Hz),

3.55-4.40 (7H, m), 5.43 (1H, dd, J = 7.6Hz, 5.4Hz), 6.73 (1H, s),

7.10-7.40 (15H, m).

Example 12

(S)-3-[(2S,3R,4R,5S)-2,3,4,6-tetrahydroxy-5-(L-leucyl)aminocaproyl]amino-3-phenylpropanoic acid ester

To (S)-3-[(2S,3R,4R,5S)-2,3,4,6-tetrahydroxy-5-(L-leucyl)aminocaproyl]amino-3-phenylpropanoic acid (Compound 3) (500 mg) was added with 28% hydrogen chloride in ethanol (200 ml), and the mixture was stirred at room temperature for 20 hours. After concentration, the residue was subjected to silica gel column chromatography and eluted with acetonitrile-water (5:1). Effective fractions were combined and concentrated under reduced pressure. The residue was dissolved in water (10 mL) and neutralized with aqueous sodium bicarbonate. The solution was passed through a DIAION HP-20SS column (Mitsubishi Kasei Co.), and then eluted with water-acetonitrile. Effective fractions were combined and concentrated under reduced pressure. The residue was recrystallized from chloroform-diethyl ether to obtain the title compound (123 mg).

¹ H-NMR (DMSO-d ₆ ) δ: 0.85-1.00 (6H, m),

1.07(3H, t, J=6.8Hz), 1.40-1.80(3H, m), 2.70-3.00(2H, m),

3.00-5.50 (8H, m), 3.96 (2H, q, J=6.8Hz), 7.10-7.45 (5H, m),

8.00-8.30 (2H, m).

Example 13

(S)-3-[(2S,3R,4R,5S)-5-(N-Benzyloxycarbonyl-L-leucyl)amino-2,3,4,6-tetrahydroxyhexanoyl]amino-3 -Phenylpropionic acid

To (S)-3-[(2S,3R,4R,5S)-2,3,4,6-tetrahydroxy-5-(L-leucyl)aminocaproyl]amino-3-phenylpropanoic acid To a mixture of (compound 3) (2.28 g) and 0.2N aqueous sodium hydroxide solution (25 ml) were added benzyl chloroformate (0.714 ml) and 1N aqueous sodium hydroxide solution (0°C). After stirring at room temperature for 3 hours, the mixture was washed with ether and acidified with 1N hydrochloric acid (20 mL). The aqueous solution was extracted with ethyl acetate, and the organic layer was washed with saturated brine, and dried over anhydrous sodium sulfate. Removal of the organic solvent gave a residue, which was recrystallized from ether-hexane to give the title compound (1.51 g).

¹ H-NMR (DMSO-d ₆ ) δ: 0.86 (3H, d, J = 6.0 Hz),

0.87(3H, d, J=6.0Hz), 1.30-1.80(3H, m), 2.60-3.00(2H, m),

3.20-5.40(8H, m), 5.04(2H, s), 7.10-7.60(11H, m),

8.16(1H, d, J=8.8Hz).

Example 14

(S)-3-[(2S,3R,4R,5S)-5-(N-Benzyloxycarbonyl-L-leucyl)amino-2,3,4,6-tetrahydroxyhexanoyl]amino-3 -Diphenylmethyl phenylpropionate

Under ice-cooling, to (S)-3-[(2S,3R,4R,5S)-2,3,4,6-tetrahydroxy-5-(L-leucyl)aminocaproyl]amino-3- To a solution of phenylpropanoid hydrochloride (4.35 g) in methanol (100 ml) was added diphenyldiazomethane (3.40 g) in methanol (100 ml), and the mixture was stirred at room temperature for 15 hours. Removal of the organic solvent gave a residue which was suspended in water (200 mL). Sodium bicarbonate (2.20 g) and benzyl chloroformate (18 ml) were added to the suspension, and the mixture was stirred at room temperature for 2 hours. The reaction mixture was extracted with ethyl acetate, and the extract was washed with saturated brine. The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was recrystallized from ethyl acetate to obtain the title compound (5.95 g).

¹ H-NMR (CD ₃ OD) δ: 0.91-0.97 (6H, m), 1.55-1.78 (3H, m),

3.03-3.10(2H,m), 3.63-3.73(3H,m),

3.91 (1H, dd, J = 9.6Hz, 1.4Hz), 4.16-4.23 (1H, d, J = 1.4Hz),

5.10(2H, s), 5.44(1H, t, J=6.2Hz), 7.13-7.36(20H, m).

Example 15

(S)-3-[(2S,3R,4R,5S)-5-(N-Benzyloxycarbonyl-L-leucyl)amino-2,3,4,6-tetrahydroxyhexanoyl]amino-3 -Phenylpropionic acid

(S)-3-[(2S,3R,4R,5S)-5-(N-benzyloxycarbonyl-L-leucyl)amino-2,3,4,6-tetrahydroxyhexanoyl]amino- A solution of diphenylmethyl 3-phenylpropanoate (1.5 g) in trifluoroacetic acid (100 ml) was stirred at room temperature for 3 hours and concentrated under reduced pressure. The residue was passed through a silica gel column and eluted with ethyl acetate:methanol (1:1). Effective fractions were combined and concentrated under reduced pressure. Recrystallization from ethyl acetate gave the title compound (0.86 g), which had the same ¹ H-NMR as in Example 13.

Example 16

(S)-3-[(2S,3R,4R,5S)-5-(N-Benzyloxycarbonyl-L-leucyl)amino-2,3,4,6-tetrahydroxyhexanoyl]amino-3 -pivaloyloxymethyl phenylpropionate

To (S)-3-[(2S,3R,4R,5S)-5-(N-benzyloxycarbonyl-L-leucyl)amino-2,3,4,6-tetrahydroxyhexanoyl]amino- To a solution of 3-phenylpropionic acid (295 mg) and 1,8-diazabicyclo[5.4.0]undec-7-ene (0.082 mL) in dimethylformamide (2 mL) was added pivaloyl Iodomethyl ester (139 mg) was dissolved in dimethylformamide (1 ml), and the mixture was stirred at room temperature for 1 hour. After adding water, the reaction mixture was extracted with ethyl acetate, and the extract was washed with 10% aqueous citric acid, saturated aqueous sodium bicarbonate and saturated brine, respectively. The organic solution was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was passed through a silica gel column and eluted with ethyl acetate:methanol (10:1). Effective fractions were combined and concentrated under reduced pressure. Recrystallization from chloroform gave the title compound (160 mg)

¹ H-NMR (DMSO-d ₆ ) δ: 0.85 (3H, d, J = 6.2 Hz),

0.86(3H, d, J=6.2Hz), 1.06(9H, s), 1.30-1.80(3H, m), 2.90-

3.05(2H, m), 3.20-5.40(8H, m), 5.04(2H, s), 5.61(2H, s),

7.10-7.60 (7H, m).

Example 17

(S)-3-[(2S,3R,4R,5S)-2,3,4,6-tetrahydroxy-5-(L-leucyl)aminocaproyl]amino-3-phenylpropanoic acid valeryloxymethyl ester

To (S)-3-[(2S,3R,4R,5S)-5-(N-benzyloxycarbonyl-L-leucyl)amino-2,3,4,6-tetrahydroxyhexanoyl]amino- To a solution of pivaloyloxymethyl 3-phenylpropanoate (150 mg) in methanol (5 ml) was added 10% palladium/activated carbon (30 mg) and the mixture was stirred at room temperature under a hydrogen atmosphere for 1.5 hr. After filtering off the catalyst, the filtrate was concentrated under reduced pressure. Recrystallization from ethyl acetate-hexane gave the title compound (101 mg).

¹ H-NMR (DMSO-d ₆ ) δ: 0.86 (3H, d, J = 5.8 Hz),

0.89(3H, d, J=5.8Hz), 1.07(9H, s), 1.10-1.90(3H, m),

2.97(2H, d, J=6.2Hz), 3.20-5.40(8H, m), 5.64(2H, s), 7.15-

7.45(5H, m), 7.84(1H, d, J=8.8Hz), 8.14(1H, d, J=9.2Hz).

Example 18

(S)-3-[(2S,3R,4R,5S)-5-(N-Benzyloxycarbonyl-L-leucyl)amino-2,3,4,6-tetrahydroxyhexanoyl]amino-3 -Phenylpropanamide

(S)-3-[(2S,3R,4R,5S)-5-(N-benzyloxycarbonyl-L-leucyl)amino-2,3,4,6-tetrahydroxyhexanoyl]amino- A solution of 3-phenylpropionic acid (295 mg), N-hydroxysuccinimide (58 mg) and N, N'-dicyclohexylcarbodiimide (103 mg) in acetonitrile (10 ml) was stirred at room temperature for 3 hours and the insoluble solid was filtered off. After removing the organic solvent, the obtained residue was dissolved in dimethylformamide (5 mL), and then 25% aqueous ammonia solution (1 mL) was added. The mixture was stirred at room temperature for 1 hour and concentrated under reduced pressure. The residue was extracted with ethyl acetate, and the extract was washed with 1N hydrochloric acid, saturated aqueous sodium bicarbonate and saturated brine, respectively. After drying over anhydrous sodium sulfate, the organic solvent was removed to give a residue, which was recrystallized from methanol-ether to give the title compound (97 mg).

¹ H-NMR (DMSO-d ₆ ) δ: 0.75-0.90 (6H, m), 0.95-1.80 (3H, m),

2.45-2.75(2H, m), 3.25-5.65(8H, m), 7.15-7.60(10H, m).

Example 19

(S)-3-[(2S,3R,4R,5S)-2,3,4,6-tetrahydroxy-5-(L-leucyl)aminocaproyl]amino-3-phenylpropanamide

To (S)-3-[(2S,3R,4R,5S)-5-(N-benzyloxycarbonyl-L-leucyl)amino-2,3,4,6-tetrahydroxyhexanoyl]amino- To a solution of 3-phenylpropanamide (200 mg) in methanol (10 ml) was added 10% palladium/activated carbon (50 mg) and the mixture was stirred at room temperature under hydrogen atmosphere for 20 hr. After filtering off the catalyst, the filtrate was concentrated under reduced pressure. Recrystallization from methanol-ether gave the title compound (140 mg).

¹ H-NMR (DMSO-d ₆ ) δ: 0.87 (3H, d, J = 6.0 Hz),

0.89(3H, d, J=6.0Hz), 1.10-2.20(3H, m), 2.40-2.80(2H, m),

3.10-5.40 (8H, m), 7.15-7.50 (5H, m), 7.79 (1H, d, J=7.8Hz),

8.34(1H, d, J=8.4Hz).

Example 20

(S)-3-[(2S,3R,4R,5S)-5-amino-2,3,4,6-tetrahydroxyhexanoyl]amino-3-phenylpropanoic acid diphenylmethyl ester hydrochloride

To (S)-3-[(2S,3R,4R,5S)-5-amino-2,3,4,6-tetrahydroxyhexanoyl]amino-3-phenylpropanoic acid (compound 5) (3.40 g ) in 1N hydrochloric acid (11 ml) was added methanol (10 ml) and the mixture was concentrated under reduced pressure. The residue was dissolved in methanol (100 ml), and diphenyldiazomethane (3.88 g) was added. The mixture was stirred at room temperature for 1.5 hours and concentrated under reduced pressure. The residue was washed with ether to give the title compound (5.36 g).

¹ H-NMR (DMSO-d ₆ ) δ: 3.07 (1H, d, J=7.0Hz), 3.50-5.80 (6H, m),

6.68(1H, s), 7.10-7.20(15H, m), 8.24(1H, d, J=8.8Hz).

Example 21

(S)-3-[(2S,3R,4R,5S)-2,3,4,6-tetrahydroxy-5-(L-ornithyl)aminocaproyl]amino-3-phenylpropionic acid di Hydrochloride

To a solution of ^Nα -benzyloxycarbonyl- ^Nδ -tert-butoxycarbonyl-L-ornithine (550 mg) in dimethoxyethane (7.5 mL) was added N-hydroxysuccinimide (173 mg) and N,N'-dicyclohexylcarbodiimide (309 mg) at 0°C and the mixture was incubated at 0°C for 24 hours. The insoluble matter formed was filtered off, and the filtrate was concentrated under reduced pressure. The residue was dissolved in dimethylformamide (5 ml), then triethylamine (0.209 ml) and (S)-3-[(2S,3R,4R,5S)-5-amino-2,3 , 4,6-Tetrahydroxyhexanoyl]amino-3-phenylpropanoic acid diphenylmethyl ester hydrochloride (818 mg) and stirred at room temperature for 72 hours. To the mixture was added 10% aqueous citric acid and the whole was extracted with ethyl acetate. The extract was washed with saturated aqueous sodium bicarbonate solution and saturated brine, respectively, and dried over anhydrous sodium sulfate. After removing the organic solvent, the obtained residue was purified by flash silica gel column chromatography, and then eluted with methanol-ethyl acetate (1:20). Effective fractions were combined and concentrated under reduced pressure. To the residue was added 4N hydrogen chloride in ethyl acetate (10 ml) and the mixture was stirred at room temperature for 2 hr. After concentration, the residue was dissolved in methanol (20 mL) and stirred with 10% palladium/activated carbon (200 mg) at room temperature under hydrogen atmosphere for 4 hours. After filtration, 1N hydrochloric acid (50 mL) was added to the filtrate and the whole was washed with ether. The aqueous layer was concentrated under reduced pressure to obtain a residue. Recrystallization from methanol-acetonitrile gave the title compound (376 mg).

¹ H-NMR (DMSO-d ₆ ) δ: 1.50-2.30 (4H, m), 2.76-6.00 (12H, m),

7.10-7.50(5H, m), 8.10-8.50(2H, m).

Example 22

(S)-3-[(2S,3R,4R,5S)-5-(α-L-glutamyl)amino-2,3,4,6-tetrahydroxyhexanoyl]amino-3-phenylpropane Hydrochloride

Following the same method described in Example 21 above, substituting N ^α -benzyloxycarbonyl-L-glutamic acid γ-tert-butyl ester for N ^α -benzyloxycarbonyl-N ^δ -tert-butoxycarbonyl-L-ornithine, Preparation of the title compound.

¹ H-NMR (DMSO-d ₆ ) δ: 1.80-2.30 (4H, m), 2.76-3.00 (2H, m),

3.50-6.00(8H, m), 7.10-7.50(5H, m).

Example 23

(S)-3-[(2S,3R,4R,5S)-2,3,4,6-tetrahydroxy-5-(O-methyl-L-seryl)aminocaproyl]amino-3- Phenylpropionic acid

At 0° C., N-hydroxysuccinimide (173 mg) and N , N'-dicyclohexylcarbodiimide (309 mg) and the mixture was incubated at 0°C for 24 hours. The formed insoluble matter was filtered off and the filtrate was concentrated under reduced pressure. The residue was dissolved in dimethylformamide (5 ml), then triethylamine (0.209 ml) and (S)-3-[(2S,3R,4R,5S)-5-amino-2,3 , 4,6-Tetrahydroxyhexanoyl]amino-3-phenylpropanoic acid diphenylmethyl ester hydrochloride (818 mg) and stirred at room temperature for 72 hours. To the mixture was added 10% aqueous citric acid and the whole was extracted with ethyl acetate. The extract was washed with saturated aqueous sodium bicarbonate solution and saturated brine, respectively, and dried over anhydrous sodium sulfate. After removing the organic solvent, the obtained residue was purified by flash silica gel column chromatography, and then eluted with methanol-ethyl acetate (1:20-1:3). Effective fractions were combined and concentrated under reduced pressure. The residue was dissolved in methanol (20 mL) and stirred with 10% palladium/charcoal (200 mg) at room temperature under hydrogen atmosphere for 2 hours. After filtration, water was added to the filtrate and the whole was washed with ether. The aqueous layer was concentrated under reduced pressure to obtain a residue. Recrystallization from methanol-ether gave the title compound (263 mg).

¹ H-NMR (DMSO-d ₆ ) δ: 2.60-2.90 (2H, m), 3.26 (3H, s), 3.20-

5.40(8H, m), 7.10-7.50(5H, m), 7.50-8.40(2H, m).

Example 24

(S)-3-[(2S,3R,4R,5S)-5-(O-benzyl-L-seryl)amino-2,3,4,6-tetrahydroxyhexanoyl]amino-3- Phenylpropionic acid

To a solution of O-benzyl-N-benzyloxycarbonyl-L-serine (329 mg) in acetonitrile (5 mL) was added N-hydroxysuccinimide (115 mg) and N,N'-bicyclo Hexylcarbodiimide (206 mg) and the mixture was stirred at room temperature for 3 hours. The insoluble matter formed was filtered off, and the filtrate was concentrated under reduced pressure. The residue was dissolved in dimethylformamide (5 ml), then triethylamine (0.139 ml) and (S)-3-[(2S,3R,4R,5S)-5-amino-2,3 , 4,6-Tetrahydroxyhexanoyl]amino-3-phenylpropanoic acid diphenylmethyl ester hydrochloride (545 mg) and stirred at room temperature for 15 hours. To the mixture was added 10% aqueous citric acid and the whole was extracted with ethyl acetate. The extract was washed with saturated aqueous sodium bicarbonate solution and saturated brine, respectively, and dried over anhydrous sodium sulfate. After removing the organic solvent, the obtained residue was purified by flash silica gel column chromatography, and then eluted with methanol-ethyl acetate (1:20). Effective fractions were combined and concentrated under reduced pressure. The residue was dissolved in methanol (30 mL) and stirred with palladium hydroxide/charcoal (100 mg) at room temperature under hydrogen atmosphere (3-4 atm) for 6 hours. After filtration, the filtrate was concentrated under reduced pressure, and the resulting residue was passed through a DIAION CHP-20P column (Mitsubishiikasei Co.), and then eluted with water-acetonitrile. Effective fractions were combined and concentrated under reduced pressure. The residue was recrystallized from methanol-ether to give the title compound (97 mg).

¹ H-NMR (DMSO-d ₆ ) δ: 2.55-2.90 (2H, m), 3.20-5.40 (10H, m),

4.50(2H, s), 7.10-7.50(10H, m), 7.84(1H, d, J=8.8Hz),

8.29(1H, d, J=11.4Hz).

Example 25

(S)-3-[(2S,3R,4R,5S)-5-(O-tert-butyl-N-fluorenylmethoxycarbonyl-L-seryl)amino-2,3,4,6 -Diphenylmethyl-tetrahydroxyhexanoyl]amino-3-phenylpropanoate

To a solution of O-tert-butyl-N-fluorenylmethoxycarbonyl-L-serine (575 mg) in acetonitrile (7.5 ml) at room temperature was added N-hydroxysuccinimide (173 mg) and N,N '-Dicyclohexylcarbodiimide (309 mg), and the mixture was stirred at room temperature for 3 hours. The insoluble solid formed was filtered off, and the filtrate was concentrated under reduced pressure. The residue was dissolved in dimethylformamide (5 ml), then triethylamine (0.139 ml) and (S)-3-[(2S,3R,4R,5S)-5-amino-2,3 , 4,6-Tetrahydroxyhexanoyl]amino-3-phenylpropanoic acid diphenylmethyl ester hydrochloride (545 mg) and stirred at room temperature for 24 hours. To the mixture was added 10% aqueous citric acid and the whole was extracted with ethyl acetate. The extract was washed with saturated aqueous sodium bicarbonate solution and saturated brine, respectively, and dried over anhydrous sodium sulfate. The organic solvent was removed, and the obtained residue was recrystallized from ether-hexane to obtain the title compound (1.156 g).

¹ H-NMR (DMSO-d ₆ ) δ: 1.19 (9H, s), 2.86-6.00 (15H, m),

6.80(1H, s), 7.05-8.20(23H, m).

Example 26

(S)-3-[(2S,3R,4R,5S)-5-(O-tert-butyl-L-seryl)amino-2,3,4,6-tetrahydroxyhexanoyl]amino-3 -Diphenylmethyl phenylpropionate

To (S)-3-[(2S,3R,4R,5S)-5-(O-tert-butyl-N-fluorenylmethoxycarbonyl-L-seryl)amino-2,3,4,6 -Diphenylmethyl-tetrahydroxyhexanoyl]amino-3-phenylpropanoate (874 mg) was added piperidine (5 ml) and the mixture was stirred at room temperature for 5 hours. Removal of the organic solvent gave a residue, which was purified by flash column chromatography on silica gel, and then eluted with methanol-ethyl acetate (1:2). Effective fractions were combined and concentrated under reduced pressure. The residue was recrystallized from ether-hexane to obtain the title compound (539 mg).

¹ H-NMR (DMSO-d ₆ ) δ: 1.12 (9H, s), 3.05 (3H, d, J=7.0Hz), 3.20-

5.40(8H, m), 6.67(1H, s), 7.10-7.40(15H, m),

7.74(1H, d, J=8.0Hz), 8.15(1H, d, J=9.0Hz).

Example 27

(S)-3-[(2S,3R,4R,5S)-2,3,4,6-tetrahydroxy-5-(L-seryl)aminocaproyl]amino-3-phenylpropanoic acid

To (S)-3-[(2S,3R,4R,5S)-5-(O-tert-butyl-L-seryl)amino-2,3,4,6-tetrahydroxyhexanoyl]amino- To diphenylmethyl 3-phenylpropanoate (300 mg) was added 4N hydrogen chloride in ethyl acetate (10 ml) and the mixture was stirred at room temperature for 1.5 hours. The organic solvent was removed to give a residue which was extracted with water. The aqueous layer was washed with ether and concentrated under reduced pressure. The residue was recrystallized from methanol-ether to obtain the title compound (208 mg).

¹ H-NMR (DMSO-d ₆ ) δ: 2.65-3.00 (2H, m), 3.20-5.60 (10H, m),

7.15-7.50(5H, m).

Example 28

(S)-3-[(2S,3R,4R,5S)-2,3,4,6-tetrahydroxy-5-(L-isoleucyl)aminocaproyl]amino-3-phenylpropanoic acid

To a solution of N-benzyloxycarbonyl-L-isoleucine (1114 mg) in acetonitrile (20 ml) was added N-hydroxysuccinimide (506 mg) and N,N'-dicyclohexylcarbohydrate at room temperature Diimine (867 mg), and the mixture was stirred at room temperature for 3 hours. The insoluble solid formed was filtered off, and the filtrate was added to (S)-3-[(2S,3R,4R,5S)-5-amino-2,3,4,6-tetrahydroxyhexanoyl]amino-3-phenyl A mixture of propionic acid (compound 5) (1369 mg) and triethylamine (0.558 mL) in dimethylformamide (100 mL). The mixture was stirred at room temperature for 96 hours and concentrated under reduced pressure. The residue was dissolved in methanol (20 mL) and stirred with 10% palladium/charcoal (1.0 g) at room temperature under hydrogen atmosphere for 24 hours. After adding 1N hydrochloric acid, the whole was filtered, and the filtrate was concentrated under reduced pressure. The residue was passed through a DIAION HP-20SS column (Mitsubishi Kasei Co.), and then eluted with water-acetonitrile. Effective fractions were combined and concentrated under reduced pressure. The residue was recrystallized from methanol-ether to obtain the title compound (199 mg).

¹ H-NMR (DMSO-d ₆ ) δ: 0.70-0.95 (6H, m), 1.20-1.90 (3H, m),

2.50-3.00(2H, m), 3.10-4.20(7H, m), 5.10-5.30(1H, m),

7.10-7.40 (5H, m), 7.90 (1H, d, J=8.0Hz),

8.30(1H, d, J=8.0Hz).

Example 29

(S)-3-[(2S,3R,4R,5S)-2,3,4,6-tetraacetoxy-5-(N-benzyloxycarbonyl-L-leucyl)aminocaproyl]amino -Diphenylmethyl 3-phenylpropionate

To (S)-3-[(2S,3R,4R,5S)-5-(N-benzyloxycarbonyl-L-leucyl)amino-2,3,4,6-tetrahydroxyhexanoyl]amino- To a solution of diphenylmethyl 3-phenylpropanoate (200 mg) in pyridine (5 ml) was added acetic anhydride (3 ml) and the mixture was stirred at room temperature for 4 days. After concentration under reduced pressure, the residue was dissolved in ethyl acetate (100 ml) and washed with 1N hydrochloric acid, saturated aqueous sodium bicarbonate and saturated brine, respectively. The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by silica gel column chromatography, and then eluted with ethyl acetate-hexane (1:1). Effective fractions were combined and concentrated under reduced pressure to give the title compound (85 mg).

¹ H-NMR (CDCl ₃ ) δ: 0.93-0.95 (6H, m), 1.51-1.78 (3H, m),

1.87(3H,s), 1.99(3H,s), 2.07(3H,s), 2.16(3H,s),

2.81(1H, dd, J=16.1Hz, 5.6Hz), 3.12(1H,dd, J=16.1Hz, 4.4Hz),

3.85(1H, dd, J=11.4Hz, 6.6Hz), 4.05(1H,dd, J=11.4Hz, 6.6Hz),

4.49(1H, m), 5.02(1H, m), 5.10(2H, s), 5.23(1H, d, J=1.8Hz),

5.33(1H, dd, J=10.0Hz, 1.8Hz), 5.41(1H, m), 5.51(1H, m),

6.36(1H, m), 6.76(1H, s), 7.01-7.34(21H, m).

Example 30

(S)-3-[(2S,3R,4R,5S)-5-(L-alanyl)amino-2,3,4,6-tetrahydroxyhexanoyl]amino-3-phenylpropanoic acid

To a solution of N-benzyloxycarbonyl-L-alanine (234 mg) in acetonitrile (30 mL) was added N-hydroxysuccinimide (123 mg) and N,N'-dicyclohexylcarbodisulfide at room temperature. imine (217 mg) and the mixture was stirred at room temperature for 3 hours. The insoluble matter formed was filtered off, and the filtrate was added to (S)-3-[(2S,3R,4R,5S)-5-amino-2,3,4,6-tetrahydroxyhexanoyl]amino-3-phenyl A solution of propionic acid (compound 5) (342 mg) and triethylamine (0.139 ml) in dimethylformamide (100 ml). The mixture was stirred at room temperature for 20 hours and concentrated under reduced pressure. 1N Hydrochloric acid was added to the residue and the whole was extracted with ethyl acetate-acetonitrile. The organic layer was washed with saturated brine and dried over anhydrous sodium sulfate. The organic solvent was removed to give a residue which was dissolved in methanol (30 mL) and stirred with 10% palladium/charcoal (200 mg) at room temperature under an atmosphere of hydrogen for 18 hours. After filtration, the filtrate was concentrated under reduced pressure, and the resulting residue was passed through a DIAIONHP-20ss column (Mitsubishii Kasei Co.), and then eluted with water-acetonitrile. Effective fractions were combined and concentrated under reduced pressure. The residue was recrystallized from methanol-ether to give the title compound (312 mg).

¹ H-NMR (DMSO-d ₆ ) δ: 1.20 (3H, d, J = 7.0 Hz), 2.60-

2.80(2H, m), 3.30-4.20(6H, m), 5.10-5.30(1H, m), 7.70-

7.90(1H, m), 8.30-8.50(1H, m).

Example 31

(S)-3-[(2S,3R,4R,5S)-2,3,4,6-tetrahydroxy-5-(L-valyl)aminocaproyl]amino-3-phenylpropanoic acid

To N-benzyloxycarbonyl-L-valine (460 mg) and (S)-3-[(2S,3R,4R,5S)-5-amino-2,3,4,6-tetrahydroxyhexanoyl Diethyl cyanophosphonate (298 mg) and triethylamine (0.191 ml) were added to a solution of diphenylmethyl amino-3-phenylpropanoate (500 mg) in dimethylformamide (50 ml) And the whole was stirred at room temperature for 15 hours. After concentration under reduced pressure, 0.1N hydrochloric acid (50 ml) was added to the residue and the whole was extracted with ethyl acetate (100 ml). The organic layer was washed with aqueous sodium bicarbonate solution and saturated brine, and dried over anhydrous sodium sulfate. The organic solvent was removed to give a residue which was dissolved in methanol (20 mL) and stirred with 10% palladium/charcoal (150 mg) at room temperature under hydrogen atmosphere for 1 hour. After filtration, the filtrate was concentrated under reduced pressure, and the resulting residue was passed through a DIAION HP-20SS column (Mitsubishii Kasei Co.), and then eluted with water-acetonitrile. Effective fractions were combined and concentrated under reduced pressure. The residue was recrystallized from methanol-ethyl acetate to obtain the title compound (663 mg).

¹ H-NMR (D ₂ O) δ: 0.85-1.10 (6H, m), 2.10 (1H, m),

2.65(2H, d, J=6.9Hz), 3.54-3.81(5H, m), 4.19-4.26(2H, m),

5.07(1H, t, J=6.9Hz), 7.24(5H, br s).

Example 32

(S)-3-[(2S,3R,4R,5S)-5-((S)-2-aminopentanoyl)amino-2,3,4,6-tetrahydroxyhexanoyl]amino-3-benzene Propionic acid

The title compound was prepared in the same manner as described in Example 31, substituting (S)-2-(benzyloxycarbonylamino)pentanoic acid for N-benzyloxycarbonyl-L-valine.

¹ H-NMR (D ₂ O) δ: 0.79 (3H, t, J=6.8Hz), 1.23 (2H, m),

1.73(2H, m), 2.61(2H, d, J=7.0Hz), 3.53-3.92(5H, m), 4.17-

4.24(2H, m), 5.06(1H, t, J=7.0Hz), 7.17-7.23(5H, m).

Example 33

(S)-3-[(2S,3R,4R,5S)-5-((S)-2-aminobutyryl)amino-2,3,4,6-tetrahydroxyhexanoyl]amino-3-benzene propionyl hydrochloride

To a solution of (S)-2-(benzyloxycarbonylamino)butanoic acid (500 mg) in acetonitrile (10 mL) was added N-hydroxysuccinimide (291 mg) and N,N'-bicyclo Hexylcarbodiimide (561 mg) and the mixture was stirred at room temperature for 3 hours. The insoluble matter formed was filtered off, and the filtrate was added to (S)-3-[(2S,3R,4R,5S)-5-amino-2,3,4,6-tetrahydroxyhexanoyl]amino-3-phenyl Propionic acid (Compound 5) (722 mg) and triethylamine (0.558 mL) in dimethylformamide (50 mL). The mixture was stirred at room temperature for 18 hours and concentrated under reduced pressure. To the residue was added 1N hydrochloric acid (50 mL) and the whole was extracted with ethyl acetate-tetrahydrofuran (1:1, 50 mL×3). The organic layer was washed with saturated brine and dried over anhydrous sodium sulfate. The organic solvent was removed to give a residue which was dissolved in methanol (50 mL) and stirred with 10% palladium/charcoal (200 mg) at room temperature under hydrogen atmosphere for 1 hour. After filtration, the filtrate was concentrated under reduced pressure, and the resulting residue was dissolved in 0.1N hydrochloric acid (0.1 ml) and passed through a DIAION HP-20SS column (Mitsubishii Kasei Co.), followed by elution with water-acetonitrile. Effective fractions were combined and concentrated under reduced pressure. The residue was recrystallized from methanol-ethyl acetate to obtain the title compound (200 mg).

¹ H-NMR (CD ₃ OD) δ: 1.05 (3H, t, J=7.6Hz), 1.85-1.96 (2H, m),

2.74(2H, d, J=6.4Hz), 3.68-3.91(5H, m), 4.30-4.33(2H, m),

5.32(1H, t, J=6.4Hz), 7.24-7.42(5H, m).

Example 34

(S)-3-[(2S,3R,4R,5S)-2,3,4,6-Tetrahydroxy-5-(L-phenylalanyl)aminocaproyl]amino-3-phenylpropanyl acid

The title compound was prepared in the same manner as described in Example 33, substituting N-benzyloxycarbonyl-L-phenylalanine for (S)-2-(benzyloxycarbonylamino)butanoic acid.

¹ H-NMR (D ₂ O) δ: 2.59 (2H, d, J = 6.4 Hz),

2.94(1H,dd,J=14.0Hz,9.0Hz), 3.18(1H,dd,J=14.0Hz,5.4Hz),

3.44-3.76(4H, m), 4.09-4.21(3H, m), 5.08(1H, t, J=6.4Hz),

7.17-7.25(10H, m).

Example 35

(S)-3-[(2S,3R,4R,5S)-5-((S)-3-Acetamido-2-aminopropionyl)amino-2,3,4,6-tetrahydroxyhexanoyl] Amino-3-phenylpropionic acid

The title compound was prepared in the same manner as described in Example 33, substituting (S)-3-acetylamino-2-(benzyloxycarbonylamino)propanoic acid for (S)-2-(benzyloxycarbonylamino)butanoic acid.

¹ H-NMR (D ₂ O) δ: 1.87 (3H, s), 2.58 (2H, d, J=7.0Hz), 3.36-

3.79(6H,m), 4.04(1H,m), 4.16-4.24(2H,m),

5.08(1H, t, J=7.0Hz), 7.23(5H, br s).

Example 36

(S)-3-[(2S,3R,4R,5S)-2,3,4,6-tetrahydroxy-5-(L-prolyl)aminocaproyl]amino-3-phenylpropanoic acid

The title compound was prepared in the same manner as described in Example 33, substituting N-benzyloxycarbonyl-L-proline for (S)-2-(benzyloxycarbonylamino)butanoic acid.

¹ H-NMR (D ₂ O) δ: 1.85-1.98 (3H, m), 2.34 (1H, m),

2.58(2H, d, J=7.0Hz), 3.16-3.32(2H, m), 3.48-3.77(4H, m),

4.18-4.30(3H, m), 5.06(1H, t, J=7.0Hz), 7.21-7.26(5H, m).

Example 37

(S)-3-[(2S,3R,4R,5S)-5-((S)-2-amino-5,5,5-trifluoropentanoyl)amino-2,3,4,6-tetra Hydroxycaproyl]amino-3-phenylpropanoic acid

The title compound was prepared in the same manner as described in Example 33, substituting (S)-2-(benzyloxycarbonyl)aminobutyric acid for (S)-2-benzyloxycarbonylamino-5,5,5-trifluoropentanoic acid .

¹ H-NMR (D ₂ O) δ: 1.95-2.30 (4H, m), 2.58 (2H, d, J=6.8Hz),

3.49-3.78(4H, m), 3.98(1H, t, J=8.4Hz), 4.18-4.24(2H, m),

5.06(1H, t, J=6.8Hz), 7.21-7.24(5H, m).

Example 38

(S)-3-[(2S,3R,4R,5S)-5-((S)-2-amino-4,4,4-trifluorobutyryl)amino-2,3,4,6-tetra Hydroxycaproyl]amino-3-phenylpropanoic acid

The title compound was prepared in the same manner as described in Example 33, substituting (S)-2-(benzyloxycarbonylamino)butyric acid for (S)-2-benzyloxycarbonylamino-4,4,4-trifluorobutanoic acid .

¹ H-NMR (D ₂ O) δ: 2.74 (2H, d, J=6.8Hz), 2.87-2.99 (2H, m),

3.64-3.98(4H, m), 4.34-4.41(3H, m), 5.21(1H, t, J=6.8Hz),

7.30-7.60 (5H, m).

Example 39

(S)-3-[(2S,3R,4R,5S)-5-((S)-2-amino-3-(methylsulfonylamino)propionyl)amino-2,3,4,6-tetra Hydroxycaproyl]amino-3-phenylpropanoic acid

The title compound was prepared in the same manner as described in Example 33, substituting (S)-2-(benzyloxycarbonylamino)butanoic acid for (S)-2-benzyloxycarbonylamino-3-(methylsulfonylamino)propanoic acid .

¹ H-NMR (D ₂ O) δ: 2.70 (2H, d, J=7.0Hz), 3.06 (3H, s), 3.54-

3.89(6H, m), 4.17(1H, t, J=6.6Hz), 4.28-4.35(2H, m),

5.18(1H, t, J=7.0Hz), 7.20-7.36(5H, m).

Example 40

(S)-3-[(2S,3R,4R,5S)-5-((S)-2-Amino-5-fluoropentanoyl)amino-2,3,4,6-tetrahydroxyhexanoyl]amino -3-Phenylpropanoic acid

The title compound was prepared in the same manner as described in Example 33, substituting (S)-2-(benzyloxycarbonylamino)butanoic acid for (S)-2-benzyloxycarbonylamino-5-fluoropentanoic acid.

¹ H-NMR (D ₂ O) δ: 1.80-2.15 (3H, m), 2.30-2.55 (1H, m),

2.69(2H, d, J=6.6Hz), 3.33-3.42(2H, m), 3.60-3.87(4H, m),

4.28-4.40(3H, m), 5.17(1H, t, J=6.6Hz), 7.32-7.34(5H, m).

Example 41

(S)-3-[(2S,3R,4R,5S)-5-((S)-2-amino-3-(formylamino)propionyl)amino-2,3,4,6-tetrahydroxy Hexanoyl]amino-3-phenylpropanoic acid

The title compound was prepared in the same manner as described in Example 33, substituting (S)-2-(benzyloxycarbonylamino)butanoic acid for (S)-2-benzyloxycarbonylamino-3-(formylamino)propanoic acid.

¹ H-NMR (D ₂ O) δ: 2.70 (2H, d, J=7.0Hz), 3.59-3.98 (7H, m),

4.17-4.33 (2H, m), 5.19 (1H, t, J=7.0Hz), 7.33-7.35 (5H, m),

8.11(1H, s).

Example 42

(S)-3-[(2S,3R,4R,5S)-5-(N-tert-butoxycarbonyl-O-(4-methoxybenzyl)-L-homoselinyl)amino-2,3 , 4,6-Tetrahydroxyhexanoyl]amino-3-phenylpropanoic acid diphenylmethyl ester

Using the same method described in Example 25, replace O-tert-butyl-N-fluorenylmethoxycarbonyl-L with N-tert-butoxycarbonyl-O-(4-methoxybenzyl)-L-homoserine - Serine, to prepare the title compound.

¹ H-NMR (DMSO-d ₆ ) δ: 1.37 (9H, s), 1.60-2.10 (2H, m),

3.05(2H, d, J=6.6Hz), 3.20-5.40(12H, m), 3.73(3H, s),

6.67(1H, s), 6.80-7.40(19H, m).

Example 43

(S)-3-[(2S,3R,4R,5S)-5-(L-homoserinyl)amino-2,3,4,6-tetrahydroxyhexanoyl]amino-3-phenylpropanoic acid

At room temperature, to (S)-3-[(2S, 3R, 4R, 5S)-5-(N-tert-butoxycarbonyl-O-(4-methoxybenzyl)-L-homoselinyl)amino -2,3,4,6-tetrahydroxyhexanoyl]amino-3-phenylpropionic acid diphenylmethyl ester (200 mg) was added with 4N hydrogen chloride in ethyl acetate (10 ml) solution, the mixture was stirred at room temperature for 3 Hour. After concentration under reduced pressure, the residue was dissolved in water and washed with ether. The aqueous layer was passed through a DIAION CHP-20P column (Mitsubishi Kasei Co.), and then eluted with water-acetonitrile. Effective fractions were combined and concentrated under reduced pressure. The residue was recrystallized from methanol-ether to obtain the title compound (53 mg).

¹ H-NMR (DMSO-d ₆ ) δ: 1.40-2.00 (2H, m), 2.55-2.80 (2H, m),

3.00-5.30(9H, m), 7.10-7.40(5H, m).

Example 44

(S)-3-[(2S,3R,4R,5S)-5-((S)-2-amino-3-cyanopropionyl)amino-2,3,4,6-tetrahydroxyhexanoyl] Amino-3-phenylpropionic acid

To a solution of (S)-2-benzyloxycarbonylamino-3-cyanopropionic acid (300 mg) in acetonitrile (10 mL) was added N-hydroxysuccinimide (177 mg) and N,N' - Dicyclohexylcarbodiimide (303 mg) and the mixture was stirred at room temperature for 2 hours. The insoluble solid formed was filtered off, and (S)-3-[(2S,3R,4R,5S)-5-amino-2,3,4,6-tetrahydroxyhexanoyl]amino- A solution of 3-phenylpropanoic acid (compound 5) (503 mg) and triethylamine (0.410 ml) in dimethylformamide (30 ml). The mixture was stirred at room temperature for 18 hours, then concentrated under reduced pressure. 1N Hydrochloric acid (50 mL) was added to the residue and the whole was extracted with ethyl acetate (100 mL×2). The extract was washed with saturated brine and dried over anhydrous sodium sulfate. After removal of the organic solvent, the obtained residue was dissolved in 4N hydrogen chloride in ethyl acetate (20 ml) and stirred at room temperature for 1 hr. After concentration, the residue was passed through a DIAION HP-20SS column (Mitsubishi Kasei Co.), and then eluted with water-acetonitrile. Effective fractions were combined and concentrated under reduced pressure. The residue was recrystallized from methanol-ethyl acetate to obtain the title compound (75 mg).

¹ H-NMR (D ₂ O) δ: 2.60 (2H, d, J=6.4Hz), 3.48-3.79 (4H, m),

4.13-4.26(3H, m), 5.18(1H, t, J=6.4Hz), 7.15-7.35(5H, m).

Example 45

(S)-3-[(2S,3R,4R,5S)-2,3,4,6-tetrahydroxy-5-(L-methionyl)aminocaproyl]amino-3-phenylpropanoic acid

To a solution of N-tert-butoxycarbonyl-L-methionine (260 mg) in acetonitrile (15 ml) was added N-hydroxysuccinimide (132 mg) and N,N'-dicyclohexyl Carbodiimide (227 mg) and the mixture was stirred at room temperature for 3 hours. The insoluble solid formed was filtered off, and (S)-3-[(2S,3R,4R,5S)-5-amino-2,3,4,6-tetrahydroxyhexanoyl]amino- 3-Phenylpropanoic acid (compound 5) (356 mg), triethylamine (0.217 mL) and dimethylformamide (50 mL) solution. The mixture was stirred at room temperature for 20 hours, then concentrated under reduced pressure. To the residue was added 4N hydrogen chloride in ethyl acetate (30 ml), and the whole was stirred at room temperature for 1.5 hr. After concentration, the residue was passed through a DIAION HP-20SS column (Mitsubishi Kasei Co.), and then eluted with water-acetonitrile. Effective fractions were combined and concentrated under reduced pressure. The residue was recrystallized from methanol-ether to obtain the title compound (310 mg).

¹ H-NMR (D ₂ O) δ: 2.12 (3H, s), 2.20 (2H, m),

2.62(2H, t, J=7.3Hz), 2.74(1H, d, J=6.9Hz), 3.60-

3.94(4H, m), 4.16(1H, t, J=6.7Hz), 4.33-4.40(2H, m),

5.20(1H, t, J=6.9Hz), 7.30-7.48(5H, m).

Example 46

(S)-3-[(2S,3R,4R,5S)-2,3,4,6-tetrahydroxy-5-(S-methyl-L-cysteinyl)aminocaproyl]amino-3 -Phenylpropionic acid

The title compound was prepared in the same manner as described in Example 45, substituting N-tert-butoxycarbonyl-S-methyl-L-cysteine for N-tert-butoxycarbonyl-L-methionine.

¹ H-NMR (DMSO-d ₆ ) δ: 2.05 (3H, s), 2.56-2.86 (4H, m), 3.40-

3.80(11H, m), 4.05(1H, m), 4.13(1H, s), 5.22(1H, m), 7.19-

7.37(5H, m), 7.84(1H, d, J=8.8Hz), 8.24(1H, d, J=8.8Hz).

Example 47

(S)-3-[(2S,3R,4R,5S)-5-((S)-2-Amino-4-pentenoyl)amino-2,3,4,6-tetrahydroxyhexanoyl]amino -3-Phenylpropanoic acid

To (S)-2-tert-butoxycarbonylamino-4-pentenoic acid (220 mg) in acetonitrile (7.5 mL) was added N-hydroxysuccinimide (118 mg) and N,N '-Dicyclohexylcarbodiimide (210 mg) and the mixture was stirred at room temperature for 2.5 hours. The insoluble solid formed was filtered off, and (S)-3-[(2S,3R,4R,5S)-5-amino-2,3,4,6-tetrahydroxyhexanoyl]amino- 3-Phenylpropanoic acid (compound 5) (342 mg), triethylamine (0.28 mL) and dimethylformamide (40 mL) solution. The mixture was stirred at room temperature for 2 days, then concentrated under reduced pressure. To the residue was added 4N hydrogen chloride in ethyl acetate (20 ml), and the whole was stirred at room temperature for 2 hr. After concentration under reduced pressure, the residue was passed through a DIAION HP-20SS column (Mitsubishi Kasei Co.), and then eluted with water-acetonitrile. Effective fractions were combined and concentrated under reduced pressure, then lyophilized. The residue was passed through a Sephadex LH-20 column (Pharmacia, Sweden) and eluted with water. Effective fractions were combined and concentrated under reduced pressure. The residue was recrystallized from methanol-ethyl acetate to obtain the title compound (57 mg).

¹ H-NMR (D ₂ O) δ: 2.64-2.76 (4H, m), 3.62-

3.82(2H, m), 3.89(1H, d, J=9.8Hz), 4.12(1H, m), 4.35(2H, m),

5.20(1H, t, J=6.8Hz), 5.25-5.35(2H, m), 5.77(1H, m), 7.30-

7.46(5H, m).

Example 48

(S)-3-[(2S,3R,4R,5S)-5-(L-alanyl-L-leucyl)amino-2,3,4,6-tetrahydroxyhexanoyl]amino-3 -Phenylpropionic acid

To a solution of N-benzyloxycarbonyl-L-alanine (50 mg) in acetonitrile (2 mL) was added N-hydroxy-5-norbornene-2,3-dicarboxyimine (47 mg) and N, N'-Dicyclohexylcarbodiimide (51 mg) and the mixture was stirred at room temperature for 1 hour. The insoluble solid formed was filtered off, and (S)-3-[(2S,3R,4R,5S)-2,3,4,6-tetrahydroxy-5-(L-leucyl ) Aminocaproyl]amino-3-phenylpropanoic acid (compound 3) (100 mg) and triethylamine (0.031 ml) in dimethylformamide (10 ml). The mixture was stirred at room temperature for 16 hours and concentrated under reduced pressure. To the residue was added 1N hydrochloric acid (0.956 ml), and the whole was extracted with ethyl acetate. The organic layer was washed with saturated brine and dried over anhydrous sodium sulfate. The organic solvent was removed to give a residue which was dissolved in methanol (10 mL) and stirred with 10% palladium/charcoal (50 mg) at room temperature under hydrogen atmosphere for 1 hour. After filtration, the filtrate was concentrated under reduced pressure, and the resulting residue was passed through a DIAION HP-20SS column (Mitsubishikasei Co.), and then eluted with water-acetonitrile. Effective fractions were combined and concentrated under reduced pressure. The residue was recrystallized from methanol-ethyl acetate to obtain the title compound (75 mg).

¹ H-NMR (CD ₃ OD) δ: 0.93-1.00 (6H, m), 1.55 (3H, d, J=7.4Hz),

1.60-1.80(3H, m), 2.68(2H, d, J=6.6Hz), 3.65-3.72(3H, m),

3.86-3.93(2H, m), 4.15-4.36(3H, m), 5.31(1H, t, J=6.6Hz),

7.22-7.41 (5H, m).

Example 49

(S)-3-[(2S,3R,4R,5S)-2,3,4,6-tetrahydroxy-5-(N-methylglycyl-L-leucyl)aminocaproyl]amino -3-Phenylpropanoic acid

The title compound was prepared in the same manner as described in Example 48, substituting N-benzyloxycarbonyl-N-methylglycine for N-benzyloxycarbonyl-L-alanine.

¹ H-NMR (CD ₃ OD) δ: 0.94 (3H, d, J = 5.8 Hz),

0.98(3H, d, J=3.2Hz), 1.60-1.80(3H, m),

2.63(2H, d, J=6.2Hz), 2.72(3H, s), 3.66-3.76(3H, m), 3.83-

3.85(3H, m), 4.14(1H, t, J=6.2Hz), 4.27-4.35(2H, m),

5.32(1H, t, J=6.2Hz), 7.20-7.40(5H, m).

Example 50

(S)-3-[(2S,3R,4R,5S)-2,3,4,6-tetrahydroxy-5-(L-phenylalanyl-L-leucyl)aminocaproyl]amino -3-Phenylpropanoic acid

The title compound was prepared in the same manner as described in Example 48, substituting N-benzyloxycarbonyl-L-phenylalanine for N-benzyloxycarbonyl-L-alanine.

¹ H-NMR (CD ₃ OD) δ: 0.94 (3H, d, J = 6.4 Hz),

0.97(3H, d, J=7.8Hz), 1.60-1.80(3H, m),

2.72 (2H, d, J = 6.6Hz), 3.05 (1H, dd, J = 14.4Hz, 8.0Hz),

3.30(1H, dd, J=14.4Hz, 4.2Hz), 3.66-3.78(3H, m),

3.91(1H, m), 4.02(1H, m), 4.22(1H, t, J=6.6Hz), 4.30-

4.36(2H, m), 5.33(1H, t, J=6.6Hz), 7.20-7.39(10H, m).

Example 51

(S)-3-[(2S,3R,4R,5S)-2,3,4,6-tetrahydroxy-5-(L-lysyl-L-leucyl)aminocaproyl]amino-3 -Phenylpropionic acid dihydrochloride

To a solution of ^Nα -benzyloxycarbonyl- ^Nε -tert-butoxycarbonyl-L-lysine (126 mg) in acetonitrile (3 mL) was added N-hydroxy-5-norbornene-2,3-di Carboxylimine (64 mg) and N,N'-dicyclohexylcarbodiimide (69 mg), the mixture was stirred at room temperature for 1 hour. The insoluble solid formed was filtered off and the filtrate was added to (S)-3-[(2S,3R,4R,5S)-2,3,4,6-tetrahydroxy-5-(L-leucyl)aminohexyl Acyl]amino-3-phenylpropionic acid (compound 3) (150 mg) and triethylamine (0.042 ml) in dimethylformamide (14 ml). The mixture was stirred at room temperature for 16 hours, then concentrated under reduced pressure. To the residue was added 1N hydrochloric acid (1.3 ml) and the whole was extracted with ethyl acetate. The extract was washed with saturated brine and dried over anhydrous sodium sulfate. The organic solvent was removed, and the resulting residue was dissolved in methanol (20 mL) and stirred with 10% palladium/charcoal (100 mg) at room temperature under hydrogen atmosphere for 2 hours. After filtration, the filtrate was concentrated under reduced pressure to obtain a solution of the residue dissolved in 4N hydrogen chloride in ethyl acetate (10 ml), and the mixture was stirred at room temperature for 2 hours. After concentration, the residue was passed through a DIAION HP-20SS column (Mitsubishi Kasei Co.), and then eluted with water-acetonitrile. Effective fractions were combined and concentrated under reduced pressure. The residue was recrystallized from methanol-ethyl acetate to obtain the title compound (180 mg).

¹ H-NMR (CD ₃ OD) δ: 0.97 (3H, d, J = 6.0 Hz),

1.00(3H, d, J=6.0Hz), 1.40-2.00(10H, m), 2.74(1H, m), 2.93-

2.99 (2H, m), 3.61-3.73 (3H, m), 3.84-4.00 (2H, m), 4.22-

4.46(3H, m), 5.37(1H, m), 7.23-7.41(5H, m).

Example 52

(S)-3-[(2S,3R,4R,5S)-5-(α-L-glutamyl-L-leucyl)amino-2,3,4,6-tetrahydroxyhexanoyl]amino -3-Phenylpropanoic acid

N-Hydroxy-5-norbornene-2,3-dicarboxyimine (64 mg ) and N, N'-dicyclohexylcarbodiimide (69 mg), and the mixture was stirred at room temperature for 1 hour. The insoluble solid formed was filtered off and the filtrate was added to (S)-3-[(2S,3R,4R,5S)-2,3,4,6-tetrahydroxy-5-(L-leucyl)aminohexyl Acyl]amino-3-phenylpropionic acid (compound 3) (150 mg) and triethylamine (0.042 ml) in dimethylformamide (14 ml). The mixture was stirred at room temperature for 16 hours, then concentrated under reduced pressure. To the residue was added 1N hydrochloric acid (1.3 ml) and the whole was extracted with ethyl acetate. The extract was washed with saturated brine and dried over anhydrous sodium sulfate. The organic solvent was removed, and the resulting residue was dissolved in methanol (20 mL) and stirred with 10% palladium/charcoal (100 mg) at room temperature under hydrogen atmosphere for 2 hours. After filtration, the filtrate was concentrated under reduced pressure to obtain a residue which was dissolved in trifluoroacetic acid (20 mL) and stirred at room temperature for 1 hour. After concentration, the residue was passed through a DIAION HP-20SS column (Mitsubishi Kasei Co.), and then eluted with water-acetonitrile. Effective fractions were combined and concentrated under reduced pressure. The residue was recrystallized from methanol-ethyl acetate to obtain the title compound (124 mg).

¹ H-NMR (CD ₃ OD) δ: 0.70 (3H, d, J = 6.2 Hz),

0.74(3H, d, J=8.0Hz), 1.38-1.60(3H, m), 1.70-2.05(2H, m),

2.21-2.35(2H, m), 2.40-2.64(2H, m), 3.44-3.47(3H, m),

3.63-3.68 (2H, m), 3.97 (1H, t, J=7.6Hz), 4.09-4.15 (2H, m),

5.12(1H, m), 6.98-7.17(5H, m).

Example 53

(S)-3-[(2S,3R,4R,5S)-5-(N-(4-aminobutyryl)-L-leucyl)amino-2,3,4,6-tetrahydroxyhexanoyl ]amino-3-phenylpropionic acid

To a solution of 4-(benzyloxycarbonylamino)butanoic acid (78 mg) in acetonitrile (10 mL) was added N-hydroxysuccinimide (41 mg) and N,N'-dicyclohexylcarbodiimide (71 mg), and the mixture was stirred at room temperature for 3 hours. The insoluble solid formed was filtered off and the filtrate was added to (S)-3-[(2S,3R,4R,5S)-2,3,4,6-tetrahydroxy-5-(L-leucyl)aminohexyl Acyl]amino-3-phenylpropionic acid (compound 3) (150 mg) and triethylamine (0.115 ml) in dimethylformamide (30 ml). The mixture was stirred at room temperature for 2 days, then concentrated under reduced pressure. To the residue was added 1N hydrochloric acid (50 ml) and the whole was extracted with ethyl acetate. The organic layer was washed with saturated brine and dried over anhydrous sodium sulfate. The organic solvent was removed and the resulting residue was dissolved in methanol (30 mL) and stirred with 10% palladium/charcoal (70 mg) at room temperature under hydrogen atmosphere for 1 hour. After filtration, the filtrate was concentrated under reduced pressure, and the obtained residue was passed through a DIAION HP-20SS column (Mitsubishi Kasei Co.), and then eluted with water-acetonitrile. Effective fractions were combined and concentrated under reduced pressure. The residue was recrystallized from methanol-ethyl acetate to obtain the title compound (130 mg).

¹ H-NMR (CD ₃ OD) δ: 0.92 (3H, d, J = 5.8 Hz),

0.96(3H, d, J=6.0Hz), 1.61-1.80(3H, m), 1.91-2.01(2H, m),

2.32(1H, m), 2.46(1H, q, J=5.8Hz), 2.65(2H, d, J=7.0Hz),

2.92-3.04(2H, m), 3.66-3.78(3H, m), 3,89(2H, d, J=9.6Hz), 4.15(1H, t, J=6.2Hz), 4.29(2H, m) , 5.32(1H, t, J=7.0Hz), 7.20-7.40(5H, m).

Example 54

(S)-3-[(2S,3R,4R,5S)-2,3,4,6-tetrahydroxy-5-(L-ornithyl-L-leucyl)aminocaproyl]amino-3 -Phenylpropionic acid

The title compound was prepared in the same manner as described in Example 53, substituting N ^α ,N ^δ -dibenzyloxycarbonyl-L-ornithine for 4-(benzyloxycarbonylamino)butanoic acid.

¹ H-NMR (CD ₃ OD) δ: 0.92-0.98 (6H, m), 1.63-1.72 (7H, m),

2.65(2H, d, J=6.6Hz), 2.90-2.92(2H, m), 3.44(1H, m), 3.65-

3.75(3H, m), 3.88(1H, dd, J=9.8Hz, 1.8Hz),

4.19 (1H, dt, J = 6.2Hz, 1.8Hz), 4.31 (1H, d, J = 1.2Hz),

4.45(1H, t, J=7.4Hz), 5.32(1H, t, J=6.6Hz), 7.20-

7.41(5H, m).

Example 55

(S)-3-[(2S,3R,4R,5S)-5-(L-asparaginyl-L-leucyl)amino-2,3,4,6-tetrahydroxyhexanoyl]amino- 3-Phenylpropanoic acid

The title compound was prepared in the same manner as described in Example 53, substituting N-benzyloxycarbonyl-L-asparagine for 4-(benzyloxycarbonylamino)butanoic acid.

¹ H-NMR (CD ₃ OD) δ: 0.92-0.99 (6H, m), 1.60-1.80 (3H, m),

2.69 (2H, d, J = 7.0Hz), 2.80 (1H, dd, J = 16.8Hz, 7.4Hz),

2.95(1H, dd, J=16.8Hz, 5.2Hz), 3.67-3.75(3H, m),

3.88(1H, m), 4.07-4.22(2H, m), 4.30-4.40(2H, m),

5.53(1H, t, J=7.0Hz), 7.20-7.41(5H, m).

Example 56

(S)-3-[(2S,3R,4R,5S)-5-(L-glutaminyl-L-leucyl)amino-2,3,4,6-tetrahydroxyhexanoyl]amino- 3-Phenylpropanoic acid

The title compound was prepared in the same manner as described in Example 53, substituting N-benzyloxycarbonyl-L-glutamine for 4-(benzyloxycarbonylamino)butanoic acid.

¹ H-NMR (CD ₃ OD) δ: 0.93-1.02 (6H, m), 1.60-1.80 (3H, m),

2.10-2.20(2H,m), 2.48-2.60(2H,m), 2.91-2.98(2H,m),

3.68-4.55(8H, m), 5.42(1H, m), 7.29-7.37(5H, m).

Example 57

(S)-3-[(2S,3R,4R,5S)-5-(N-((S)-3-acetylamino-2-aminopropionyl)-L-leucyl)amino-2,3 , 4,6-Tetrahydroxyhexanoyl]amino-3-phenylpropanoic acid

The title compound was prepared in the same manner as described in Example 53, substituting (S)-3-acetylamino-2-(benzyloxycarbonylamino)propanoic acid for 4-(benzyloxycarbonylamino)butanoic acid.

¹ H-NMR (CD ₃ OD) δ: 0.93-1.00 (6H, m), 1.64-1.70 (3H, m),

1.98(3H, s), 2.70(2H, d, J=6.8Hz), 3.51-3.98(7H, m), 4.21-

4.41(3H, m), 5.33(1H, t, J=6.8Hz), 7.20-7.43(5H, m).

Example 58

(S)-3-[(2S,3R,4R,5S)-5-(N-((S)-2,3-diaminopropionyl)-L-leucyl)amino-2,3,4 , 6-Tetrahydroxyhexanoyl]amino-3-phenylpropanoic acid

The title compound was prepared in the same manner as described in Example 53, substituting (S)-2,3-(dibenzyloxycarbonylamino)propanoic acid for 4-(benzyloxycarbonylamino)butanoic acid.

¹ H-NMR (D ₂ O) δ: 0.74-0.81 (6H, m), 1.40-1.65 (3H, m),

2.59 (2H, d, J = 7.0Hz), 2.98 (1H, dd, J = 15.2Hz, 7.6Hz),

3.15(1H, dd, J=15.2Hz, 5.8Hz), 3.46-3.78(5H, m), 4.09-

4.30(3H, m), 5.06(1H, t, J=7.0Hz), 7.18-7.27(5H, m).

Example 59

(S)-3-[(2S,3R,4R,5S)-2,3,4,6-tetrahydroxy-5-((O-methyl-L-threonyl)-L-leucyl) Aminocaproyl]amino-3-phenylpropionic acid

The title compound was prepared in the same manner as described in Example 53, substituting N-benzyloxycarbonyl-O-methyl-L-threonine for 4-(benzyloxycarbonylamino)butanoic acid.

¹ H-NMR (CD ₃ OD) δ: 0.94-1.01 (6H, m), 1.15 (3H, d, J=6.6Hz),

1.62-1.73(3H, m), 2.75-3.00(2H, m), 3.43(3H, s), 3.60-

3.74(4H, m), 3.88-3.94(2H, m), 4.12-4.22(2H, m),

4.33(1H, s), 5.37(1H, t, J=6.2Hz), 7.32-7.38(5H, m).

Example 60

(S)-3-[(2S,3R,4R,5S)-5-(N-((S)-2-amino-3-cyclohexylpropionyl)-L-leucyl)amino-2,3 , 4,6-Tetrahydroxyhexanoyl]amino-3-phenylpropanoic acid

The title compound was prepared in the same manner as described in Example 53, substituting (S)-2-benzyloxycarbonylamino-3-cyclohexylpropanoic acid for 4-(benzyloxycarbonylamino)butanoic acid.

¹ H-NMR (D ₂ O) δ: 0.77-1.13 (11H, m), 1.50 (11H, m),

2.58(2H, d, J=5.8Hz), 3.54-4.22(8H, m),

5.05(1H, t, J=5.8Hz), 7.23(5H, br s).

Example 61

(S)-3-[(2S,3R,4R,5S)-5-(N-((S)-2-amino-5,5,5-trifluoropentanoyl)-L-leucyl)amino -2,3,4,6-Tetrahydroxyhexanoyl]amino-3-phenylpropanoic acid

The title compound was prepared in the same manner as described in Example 53, substituting (S)-2-dibenzyloxycarbonylamino-5,5,5-trifluoropentanoic acid for 4-(benzyloxycarbonylamino)butanoic acid.

¹ H-NMR (D ₂ O) δ: 0.75-0.81 (6H, m), 1.42-1.60 (3H, m), 1.98-

2.22(4H, m), 2.59(2H, d, J=6.6Hz), 3.43-3.59(3H, m), 3.73-

3.95(2H, m), 4.08-4.36(3H, m), 5.06(1H, t, J=6.6Hz), 7.15-

7.35(5H, m).

Example 62

(S)-3-[(2S,3R,4R,5S)-2,3,4,6-tetrahydroxy-5-(L-leucyl-L-leucyl)aminocaproyl]amino-3 -Phenylpropionic acid

The title compound was prepared in the same manner as described in Example 53, substituting N-benzyloxycarbonyl-L-leucine for 4-(benzyloxycarbonylamino)butanoic acid.

¹ H-NMR (CD ₃ OD) δ: 0.80-1.00 (12H, m), 1.00-1.90 (6H, m),

2.60-2.80(2H, m), 3.20-5.30(9H, m), 7.10-7.60(5H, m).

Example 63

(S)-3-[(2S,3R,4R,5S)-2,3,4,6-tetrahydroxy-5-(L-isoleucyl-L-leucyl)aminocaproyl]amino- 3-Phenylpropanoic acid

The title compound was prepared in the same manner as described in Example 53, substituting N-benzyloxycarbonyl-L-isoleucine for 4-(benzyloxycarbonylamino)butanoic acid.

¹ H-NMR (CD ₃ OD) δ: 0.70-1.80 (18H, m), 2.60-2.90 (2H, m),

3.00-5.30(9H, m), 7.10-7.50(5H, m).

Example 64

(S)-3-[(2S,3R,4R,5S)-2,3,4,6-tetrahydroxy-5-((O-methyl-L-seryl)-L-leucyl) Aminocaproyl]amino-3-phenylpropionic acid

The title compound was prepared in the same manner as described in Example 53, substituting N-benzyloxycarbonyl-O-methyl-L-serine for 4-(benzyloxycarbonylamino)butanoic acid.

¹ H-NMR (DMSO-d ₆ ) δ: 0.85 (3H, d, J = 6.0 Hz),

0.88(3H, d, J=6.0Hz), 1.40-1.70(3H, m), 2.70-2.80(2H, m),

3.25(3H, s), 3.10-5.30(10H, m), 7.10-7.60(5H, m), 8.00-

8.40(2H, m).

Example 65

(S)-3-[(2S,3R,4R,5S)-2,3,4,6-tetrahydroxy-5-(N-(2-phenylglycyl)-L-leucyl)amino Hexanoyl]amino-3-phenylpropanoic acid

The title compound was prepared in the same manner as described in Example 53, substituting N-benzyloxycarbonyl-2-phenylglycine for 4-(benzyloxycarbonylamino)butanoic acid.

¹ H-NMR (CD ₃ OD) δ: 0.70-1.00 (6H, m), 1.50-1.70 (3H, m),

2.60-2.80(2H, m), 3.40-5.50(9H, m), 7.10-7.60(10H, m).

Example 66

(S)-3-[(2S,3R,4R,5S)-2,3,4,6-tetrahydroxy-5-((N-methyl-L-valyl)-L-leucyl) Aminocaproyl]amino-3-phenylpropionic acid

The title compound was prepared in the same manner as described in Example 53, substituting N-benzyloxycarbonyl-N-methyl-L-valine for 4-(benzyloxycarbonylamino)butanoic acid.

¹ H-NMR (DMSO-d ₆ ) δ: 0.70-1.00 (12H, m), 1.40-1.90 (4H, m),

2.19(3H, s), 2.50-5.30(11H, m), 7.10-7.50(6H, m),

8.03(1H, d, J=9.2Hz), 8.20(1H, d, J=8.8Hz).

Example 67

(S)-3-[(2S,3R,4R,5S)-5-(N-((S)-2-aminobutyryl)-L-leucyl)amino-2,3,4,6- Tetrahydroxyhexanoyl]amino-3-phenylpropion hydrochloride

To a solution of (S)-2-(tert-butoxycarbonylamino)butanoic acid (70 mg) in acetonitrile (10 mL) was added N-hydroxysuccinimide (41 mg) and N,N'-dicyclohexyl Carbodiimide (71 mg), and the mixture was stirred at room temperature for 3 hours. The insoluble solid formed was filtered off and the filtrate was added to (S)-3-[(2S,3R,4R,5S)-2,3,4,6-tetrahydroxy-5-(L-leucyl)aminohexyl Acyl]amino-3-phenylpropionic acid (compound 3) (150 mg) and triethylamine (0.115 ml) in dimethylformamide (30 ml). The mixture was stirred at room temperature for 2 days, then concentrated under reduced pressure. To the residue was added 1N hydrochloric acid (50 ml) and the whole was extracted with ethyl acetate. The extract was washed with saturated brine and dried over anhydrous sodium sulfate. The organic solvent was removed, and the obtained residue was dissolved in 4N hydrogen chloride in ethyl acetate (10 ml) and stirred at room temperature for 2 hours. After concentration, the residue was passed through a DIAION HP-20SS column (Mitsubishi Kasei Co.), and then eluted with water-acetonitrile. Effective fractions were combined and concentrated under reduced pressure. The residue was recrystallized from methanol-ethyl acetate to obtain the title compound (108 mg).

¹ H-NMR (CD ₃ OD) δ: 0.94-1.09 (9H, m), 1.6 2-1.72 (3H, m),

1.84-1.98(2H, m), 2.69(2H, d, J=6.4Hz), 3.68-3.83(3H, m),

4.19-4.43(5H, m), 5.32(1H, t, J=6.4Hz), 7.22-7.43(5H, m).

Example 68

(S)-3-[(2S,3R,4R,5S)-2,3,4,6-tetrahydroxy-5-(L-norvalyl-L-leucyl)aminocaproyl]amino- 3-Phenylpropion hydrochloride

The title compound was prepared in the same manner as described in Example 67, substituting N-tert-butoxycarbonyl-L-norvaline for (S)-2-(tert-butoxycarbonylamino)butanoic acid.

¹ H-NMR (CD ₃ OD) δ: 0.93-1.02 (9H, m), 1.44-1.52 (2H, m),

1.62-1.74(3H, m), 1.81-1.90(2H, m), 2.69(2H, d, J=6.2Hz),

3.68-3.83(3H, m), 4.18-4.47(5H, m), 5.32(1H, t, J=6.2Hz),

7.19-7.43 (5H, m).

Example 69

(S)-3-[(2S,3R,4R,5S)-2,3,4,6-tetrahydroxy-5-(L-norleucyl-L-leucyl)aminocaproyl]amino- 3-Phenylpropion hydrochloride

The title compound was prepared in the same manner as described in Example 67, substituting N-tert-butoxycarbonyl-L-norleucine for (S)-2-(tert-butoxycarbonylamino)butanoic acid.

¹ H-NMR (CD ₃ OD) δ: 0.91-1.00 (9H, m), 1.37-1.50 (6H, m),

1.63-1.77(3H, m), 1.87-1.94(2H, m), 2.82-2.86(2H, m),

3.64-3.72(2H,m), 3.85-3.95(2H,m), 4.09-4.23(2H,m),

4.30-4.41(2H, m), 5.37(1H, t, J=6.6Hz), 7.23-7.42(5H, m).

Example 70

(S)-3-[(2S,3R,4R,5S)-5-(D-alanyl-L-leucyl)amino-2,3,4,6-tetrahydroxyhexanoyl]amino-3 -Phenylpropion hydrochloride

The title compound was prepared in the same manner as described in Example 67, substituting N-tert-butoxycarbonyl-D-alanine for (S)-2-(tert-butoxycarbonylamino)butanoic acid.

¹ H-NMR (CD ₃ OD) δ: 0.92-1.01 (6H, m), 1.51 (3H, d, J=7.0Hz),

1.58-1.73(3H, m), 2.66-2.71(2H, m), 3.60-3.74(2H, m),

3.91-4.02(2H, m), 4.09-4.36(4H, m), 5.33(1H, m), 7.25-

7.40(5H,m).

Example 71

(S)-3-[(2S,3R,4R,5S)-5-((β-cyano-L-alanyl)-L-leucyl)amino-2,3,4,6-tetra Hydroxycaproyl]amino-3-phenylpropanoic acid

The title compound was prepared in the same manner as described in Example 67, substituting N-tert-butoxycarbonyl-β-cyano-L-alanine for (S)-2-(tert-butoxycarbonylamino)butanoic acid.

¹ H-NMR (CD ₃ OD) δ: 0.93-0.99 (6H, m), 1.61-1.82 (3H, m),

2.74-2.95(4H, m), 3.62-3.92(5H, m), 4.17-4.46(3H, m),

5.37(1H, t, J=6.6Hz), 7.26-7.38(5H, m).

Example 72

N-[(S)-3-[(2S,3R,4R,5S)-2,3,4,6-tetrahydroxy-5-(L-leucyl)aminocaproyl]amino-3-phenyl propionyl]-L-alanine

To (S)-3-[(2S,3R,4R,5S)-5-(N-benzyloxycarbonyl-L-leucyl)amino-2,3,4,6-tetrahydroxyhexanoyl]amino- 3-Phenylpropanoic acid (200 mg), L-alanine benzyl p-toluenesulfonate (238 mg) and N-hydroxy-5-norbornene-2,3-dicarboxyimine (183 mg) N,N'-dicyclohexylcarbodiimide (140 mg) and triethylamine (0.094 ml) were added to a solution of acetonitrile (10 ml), and the mixture was stirred at room temperature for 15 hours. The insoluble solid formed was filtered off, and the filtrate was concentrated under reduced pressure. The residue was dissolved in ethyl acetate and washed with saturated brine, then dried over anhydrous sodium sulfate. The organic solvent was removed, and the resulting residue was dissolved in methanol (10 mL) and stirred with 10% palladium/charcoal (60 mg) at room temperature under hydrogen atmosphere for 3 hours. After filtration, the filtrate was concentrated under reduced pressure to obtain a residue, which was passed through a DIAIONHP-20SS column (Mitsubishi Kasei Co.), and then eluted with water-acetonitrile. Effective fractions were combined and concentrated under reduced pressure. The residue was recrystallized from methanol-ethyl acetate to obtain the title compound (58 mg).

¹ H-NMR (CD ₃ OD) δ: 0.99-1.01 (6H, m), 1.26 (3H, d, J=7.4Hz),

1.70-1.73(3H, m), 2.74-2.79(2H, m), 3.67-3.82(3H, m),

3.87-3.95(2H, m), 4.12-4.32(2H, m), 4.35(1H, d, J=1.2Hz),

5.39(1H, dd, J=7.0Hz, 5.4Hz), 7.20-7.39(5H, m).

Example 73

N-[(S)-3-[(2S,3R,4R,5S)-2,3,4,6-tetrahydroxy-5-(L-leucyl)aminocaproyl]amino-3-phenyl propionyl]-L-leucine

The title compound was prepared in the same manner as described in Example 72, substituting L-alanine benzyl p-toluenesulfonate for L-leucine benzyl p-toluenesulfonate.

¹ H-NMR (CD ₃ OD) δ: 0.79-1.00 (12H, m), 1.30-1.96 (6H, m),

2.78-2.81(2H, m), 3.66-3.81(3H, m), 3.85-3.92(2H, m),

4.20-4.39(3H, m), 5.38(1H, t, J=6.0Hz), 7.20-7.34(5H, m).

Example 74

(S)-3-[(2S,3R,4R,5S)-2,3,4,6-tetrahydroxy-5-((Oβ-methyl-α-L-aspartyl)-L-leu Aminoacyl)aminocaproyl]amino-3-phenylpropionic acid

To a solution of N-tert-butoxycarbonyl-L-aspartic acid β-methyl ester (194 mg) in acetonitrile (5 mL) was added N-hydroxysuccinimide (56 mg) and N,N'-di Cyclohexylcarbodiimide (95 mg), and the mixture was stirred at room temperature for 2 hours. The insoluble solid formed was filtered off, and the filtrate was added to (S)-3-[(2S,3R,4R,5S)-2,3,4,6-tetrahydroxy-5-(L-leucyl)aminocaproyl ]Amino-3-phenylpropionic acid (compound 3) (200 mg) and triethylamine (0.10 ml) in dimethylformamide (30 ml). The mixture was stirred at room temperature for 17 hours, and concentrated under reduced pressure. To the residue was added 1N hydrochloric acid (50 ml) and the whole was extracted with ethyl acetate. The organic layer was washed with saturated brine and dried over anhydrous sodium sulfate. The organic solvent was removed, and the obtained residue was dissolved in trifluoroacetic acid (20 ml) and stirred at room temperature, and concentrated under reduced pressure to obtain a residue, which was passed through a DIAION HP-20SS column (Mitsubishi kasei company), and then eluted with water-acetonitrile . Effective fractions were combined and concentrated under reduced pressure. The residue was recrystallized from methanol-ethyl acetate to obtain the title compound (92 mg).

¹ H-NMR (CD ₃ OD) δ: 0.93-1.00 (6H, m), 1.63-1.76 (3H, m),

2.74 (2H, d, J = 6.3Hz), 2.87 (1H, dd, J = 17.4Hz, 7.8Hz),

3.04(1H, dd, J=17.4Hz, 5.4Hz), 3.65-3.69(3H, m),

3.73(3H, s), 3.86(1H, dd, J=9.6Hz, 1.4Hz), 4.07(1H, m),

4.21(1H, m), 4.33-4.39(2H, m), 5.34(1H, t, J=6.3Hz), 7.20-

7.43(5H, m).

Example 75

(S)-3-[(2S,3R,4R,5S)-5-((N-Benzyloxycarbonylglycyl)-L-leucyl)amino-2,3,4,6-tetrahydroxyhexyl Acyl]amino-3-phenylpropanoic acid diphenylmethyl ester

At 0°C, add N-hydroxysuccinimide (1.15 g) and N, N'-dicyclohexyl carbonation in a solution of N-benzyloxycarbonylglycine (2.09 g) in dimethoxyethane (50 ml). Diimine (2.06 g) and the mixture was incubated at 4°C for 62 hours. The insoluble solid formed was filtered off, the filtrate was concentrated under reduced pressure, and the obtained residue was recrystallized from dichloromethane-hexane to give N-benzyloxycarbonylglycine N-hydroxysuccinimide ester (3.00 g).

N-Benzyloxycarbonylglycine N-hydroxysuccinimide ester (148 mg) was dissolved in dimethylformamide (6 ml) and (S)-3-[(2S,3R, 4R,5S)-diphenylmethyl 2,3,4,6-tetrahydroxy-5-(L-leucyl)aminocaproyl]amino-3-phenylpropanoate (300 mg). The mixture was stirred at room temperature for 1.5 hours, then 10% aqueous citric acid was added. The whole was extracted with ethyl acetate, and the extract was washed with saturated aqueous sodium bicarbonate and saturated brine, respectively, and dried over anhydrous sodium sulfate. The organic solvent was removed, and the obtained residue was recrystallized from ethyl acetate-diethyl ether to obtain the title compound (275 mg).

¹ H-NMR (CD ₃ OD) δ: 0.92 (3H, d, J = 5.8 Hz),

0.95(3H, d, J=5.8Hz), 1.20-1.80(3H, m),

2.99 (1H, dd, J = 15.8Hz, 7.4Hz), 3.10 (1H, dd, J = 15.8Hz, 6.2Hz),

3.50-4.60 (9H, m), 5.08 (2H, s), 5.43 (1H, dd, J=7.4Hz, 6.2Hz),

6.95(1H, s), 7.10-7.40(20H, m).

Example 76

(S)-3-[(2S,3R,4R,5S)-5-(Glycyl-L-leucyl)amino-2,3,4,6-tetrahydroxyhexanoyl]amino-3-benzene Propionic acid

(S)-3-[(2S,3R,4R,5S)-5-((N-Benzyloxycarbonylglycyl)-L-leucyl)amino-2,3,4,6-tetrahydroxyhexyl Acyl]amino-3-phenylpropanoic acid diphenylmethyl ester (273 mL) was dissolved in methanol (30 mL) and stirred with 10% palladium/charcoal (50 mg) at room temperature under hydrogen atmosphere for 1.5 hours. After dilution with methanol, the mixture was filtered, and the filtrate was concentrated under reduced pressure. The obtained residue was recrystallized from methanol-diethyl ether to obtain the title compound (154 mg).

¹ H-NMR (CD ₃ OD) δ: 0.94 (3H, d, J = 6.2 Hz),

0.99(3H, d, J=6.2Hz), 1.10-1.85(3H, m),

2.66(1H, d, J=7.6Hz), 3.50-4.40(9H, m),

5.33(1H, t, J=7.6Hz), 7.10-7.45(5H, m).

Example 77

(S)-3-[(2S,3R,4R,5S)-5-((N-Benzyloxycarbonyl-L-prolyl)-L-leucyl)amino-2,3,4,6- Tetrahydroxyhexanoyl]amino-3-phenylpropanoic acid diphenylmethyl ester

The title compound was prepared in the same manner as described in Example 75, substituting N-benzyloxycarbonyl-L-proline for N-benzyloxycarbonylglycine.

¹ H-NMR (DMSO-d ₆ ) δ: 0.60-2.30 (13H, m), 3.00-5.40 (14H, m),

6.68(1H, s), 7.05-7.45(20H, m).

Example 78

(S)-3-[(2S,3R,4R,5S)-2,3,4,6-tetrahydroxy-5-(L-prolyl-L-leucyl)aminocaproyl]amino-3 -Phenylpropionic acid

According to the same method described in Example 76, use (S)-3-[(2S, 3R, 4R, 5S)-5-((N-benzyloxycarbonyl-L-prolyl)-L-leucyl) Amino-2,3,4,6-tetrahydroxyhexanoyl]amino-3-phenylpropanoic acid diphenylmethyl ester instead of (S)-3-[(2S,3R,4R,5S)-5-(( Diphenylmethyl N-benzyloxycarbonylglycyl)-L-leucyl)amino-2,3,4,6-tetrahydroxyhexanoyl]amino-3-phenylpropanoate to prepare the title compound.

¹ H-NMR (DMSO-d ₆ ) δ: 0.80-1.00 (6H, m), 1.40-2.20 (7H, m),

2.60-5.30(13H, m), 7.15-7.45(5H, m), 7.59(1H, d, J=8.8Hz),

8.15-8.30 (2H, m).

Example 79

(S)-3-[(2S, 3R, 4R, 5S)-5-((O-benzyl-N-benzyloxycarbonyl-L-seryl)-L-leucyl)amino-2,3 , 4,6-Tetrahydroxyhexanoyl]amino-3-phenylpropanoic acid diphenylmethyl ester

The title compound was prepared in the same manner as described in Example 75, substituting O-benzyl-N-benzyloxycarbonyl-L-serine for N-benzyloxycarbonylglycine.

¹ H-NMR (DMSO-d ₆ ) δ: 0.85-0.95 (6H, m), 1.50-1.70 (3H, m),

2.97(1H,dd,J=14.6Hz,7.0Hz), 3.09(1H,dd,J=14.6Hz,6.6Hz),

3.60-4.50(10H, m), 4.47(1H, d, J=11.6Hz),

4.56 (1H, d, J = 11.6Hz), 5.05 (1H, d, J = 12.0Hz),

5.13 (1H, d, J = 12.0Hz), 5.43 (1H, dd, J = 7.0Hz, 6.6Hz),

6.72(1H, s), 7.10-7.40(20H, m).

Example 80

(S)-3-[(2S,3R,4R,5S)-5-((O-benzyl-L-seryl)-L-leucyl)amino-2,3,4,6-tetra Hydroxycaproyl]amino-3-phenylpropanoic acid

According to the same method described in Example 76, use (S)-3-[(2S, 3R, 4R, 5S)-5-((O-benzyl-N-benzyloxycarbonyl-L-seryl)-L -Leucyl)amino-2,3,4,6-tetrahydroxyhexanoyl]amino-3-phenylpropanoic acid diphenylmethyl ester instead of (S)-3-[(2S,3R,4R,5S) -Diphenylmethyl 5-((N-benzyloxycarbonylglycyl)-L-leucyl)amino-2,3,4,6-tetrahydroxyhexanoyl]amino-3-phenylpropanoate, Preparation of the title compound.

¹ H-NMR (CD ₃ OD) δ: 0.93 (3H, d, J = 6.6 Hz),

0.96(3H, d, J=6.6Hz), 1.05-1.80(3H, m),

2.76(2H, d, J=6.6Hz), 3.50-4.50(10H, m), 4.62(2H, s), 5.30-

5.50(1H, m), 7.15-7.50(10H, m).

Example 81

(S)-3-[(2S,3R,4R,5S)-2,3,4,6-tetrahydroxy-5-(L-seryl-L-leucyl)aminocaproyl]amino-3 -Phenylpropionic acid

To a solution of O-benzyl-N-benzyloxycarbonyl-L-serine (228 mg) in acetonitrile (20 mL) was added N-hydroxysuccinimide (83 mg) and N,N'-dicyclohexylcarbodi imine (143 mg), and the mixture was stirred at room temperature for 3 hours. The insoluble solid formed was filtered off, and (S)-3-[(2S,3R,4R,5S)-2,3,4,6-tetrahydroxy-5-(L-leucyl ) Aminocaproyl]amino-3-phenylpropanoic acid (compound 3) (300 mg) and triethylamine (0.092 ml) in dimethylformamide (60 ml). The mixture was stirred at room temperature for 21 hours and concentrated under reduced pressure. 1N Hydrochloric acid was added to the residue, and the whole was extracted with ethyl acetate. The organic layer was washed with saturated brine and dried over anhydrous sodium sulfate. The organic solvent was removed to give a residue which was dissolved in methanol (20 mL) and stirred with palladium hydroxide/charcoal (200 mg) at room temperature under an atmosphere of hydrogen (3 atm) for 10 hours. After filtration, the filtrate was concentrated under reduced pressure, and the resulting residue was passed through a DIAION HP-20SS column (Mitsubishi Kasei Co.), and then eluted with water-acetonitrile. Effective fractions were combined and concentrated under reduced pressure. The residue was recrystallized from methanol-ether to obtain the title compound (60 mg).

¹ H-NMR (CD ₃ OD) δ: 0.94 (3H, d, J = 5.8 Hz),

0.98(3H, d, J=5.8Hz), 1.60-1.85(3H, m),

2.67(2H, d, J=6.6Hz), 3.50-4.50(10H, m),

5.33(1H, t, J=6.6Hz), 7.15-7.50(5H, m).

Example 82

(S)-3-[(2S,3R,4R,5S)-5-((N-Benzyloxycarbonyl-L-valyl)-L-leucyl)amino-2,3,4,6- Tetrahydroxyhexanoyl]amino-3-phenylpropanoic acid diphenylmethyl ester

Following the same procedure described in Example 75, substituting N-benzyloxycarbonyl-L-valine for N-benzyloxycarbonylglycine, the title compound was prepared.

¹ H-NMR (DMSO-d ₆ ) δ: 0.75-0.95 (12H, m), 1.40-2.10 (4H, m),

3.04(1H, d, J=6.8Hz), 3.40-5.40(9H, m), 5.04(2H, s),

6.68(1H, s), 7.10-7.50(20H, m), 7.98(1H, d, J=7.6Hz),

8.16(1H, d, J=9.2Hz).

Example 83

(S)-3-[(2S,3R,4R,5S)-6-Acetoxy-5-((N-benzyloxycarbonyl-L-valyl)-L-leucyl)amino-2, 3,4-Trihydroxyhexanoyl]amino-3-phenylpropanoic acid diphenylmethyl ester

To (S)-3-[(2S, 3R, 4R, 5S)-5-((N-benzyloxycarbonyl-L-valyl)-L-leucyl)amino-2,3,4,6 To a solution of diphenylmethyl tetrahydroxyhexanoyl]amino-3-phenylpropanoate (200 mg) in pyridine (1 ml) was added acetic anhydride (0.0243 ml), and the mixture was stirred at room temperature for 14 hours. After concentration under reduced pressure, the residue was purified by silica gel column chromatography, and then eluted with ethyl acetate-methanol (20:1). The effective fractions were combined and concentrated under reduced pressure to obtain a residue. This was recrystallized from methanol-ether to obtain the title compound (103 mg).

¹ H-NMR (CD ₃ OD) δ: 0.80-1.05 (12H, m), 1.45-2.20 (4H, m),

2.02(3H, s), 3.03(1H, dd, J=13.4Hz, 8.4Hz),

3.21(1H, dd, J=13.4Hz, 9.6Hz), 3.60-4.60(8H, m),

5.08(2H, s), 5.40-5.50(1H, m), 6.73(1H, s), 7.10-

7.40(20H, m).

Example 84

(S)-3-[(2S,3R,4R,5S)-6-Acetoxy-2,3,4-trihydroxy-5-(L-valyl-L-leucyl)aminocaproyl ]amino-3-phenylpropionic acid

According to the same method described in Example 76, with (S)-3-[(2S, 3R, 4R, 5S)-6-acetoxy-5-((N-benzyloxycarbonyl-L-valyl)- Diphenylmethyl L-leucyl)amino-2,3,4-trihydroxyhexanoyl]amino-3-phenylpropanoate instead of (S)-3-[(2S,3R,4R,5S)- 5-((N-Benzyloxycarbonylglycyl)-L-leucyl)amino-2,3,4,6-tetrahydroxyhexanoyl]amino-3-phenylpropanoic acid diphenylmethyl ester, preparation title compound.

¹ H-NMR (CD ₃ OD) δ: 0.90-1.15 (12H, m), 1.50-2.30 (4H, m),

2.03(3H, s), 2.80(2H, d, J=6.6Hz), 5.32(1H, t, J=6.6Hz),

7.10-7.50(5H, m).

Example 85

(S)-3-[(2S,3R,4R,5S)-5-((N-Benzyloxycarbonyl-L-valyl)-L-leucyl)amino-2,3-dihydroxy-4 , 6-(O-isopropylidene)dioxyhexanoyl]amino-3-phenylpropanoic acid diphenylmethyl ester

To (S)-3-[(2S, 3R, 4R, 5S)-5-((N-benzyloxycarbonyl-L-valyl)-L-leucyl)amino-2,3,4,6 -Diphenylmethyl-tetrahydroxyhexanoyl]amino-3-phenylpropanoate (200 mg) in tetrahydrofuran (5 mL) was added 2,2-dimethoxypropane (0.288 mL) and p-toluenesulfonic acid monohydrate (4 mg), and the mixture was stirred at room temperature for 1 hour. After adding a saturated aqueous sodium bicarbonate solution, the whole was extracted with ethyl acetate, and the extract was washed with saturated brine, followed by drying over anhydrous sodium sulfate. The organic solvent was removed, and the obtained residue was recrystallized from ethyl acetate-hexane to obtain the title compound (183 mg).

¹ H-NMR (CD ₃ OD) δ: 0.80-1.10 (12H, m), 1.10-2.10 (4H, m),

1.34(3H, s), 1.44(3H, s), 2.80-3.10(2H, m), 3.30-

4.40(8H, m), 5.05(1H, d, J=12.4Hz), 5.13(1H, d, J=12.4Hz),

6.81(1H, s), 7.10-7.40(20H, m).

Example 86

(S)-3-[(2S,3R,4R,5S)-2,3-dihydroxy-4,6-(O-isopropylidene)dioxy-5-(L-valyl-L -leucyl)aminocaproyl]amino-3-phenylpropanoic acid

According to the same method described in Example 76, use (S)-3-[(2S, 3R, 4R, 5S)-5-((N-benzyloxycarbonyl-L-valyl)-L-leucyl) Amino-2,3-dihydroxy-4,6-(O-isopropylidene)dioxyhexanoyl]amino-3-phenylpropanoic acid diphenylmethyl ester instead of (S)-3-[(2S , 3R, 4R, 5S)-5-((N-benzyloxycarbonylglycyl)-L-leucyl)amino-2,3,4,6-tetrahydroxyhexanoyl]amino-3-phenylpropane acid diphenylmethyl ester to prepare the title compound.

¹ H-NMR (CD ₃ OD) δ: 0.79 (3H, d, J=6.8Hz), 0.80-0.95 (9H, m),

1.33(3H, s), 1.40(3H, s), 1.30-2.10(4H, m), 2.55-

2.90 (3H, m), 3.00-5.50 (11H, m), 7.10-7.40 (5H, m), 8.00-

8.45(3H, m).

Example 87

(S)-3-[(2S,3R,4R,5S)-2,3,4,6-tetrahydroxy-5-(L-tryptophanyl-L-leucyl)aminocaproyl]amino-3 -Phenylpropionic acid

To a solution of N-benzyloxycarbonyl-L-tryptophan (355 mg) in acetonitrile (30 mL) was added N-hydroxysuccinimide (127 mg) and N,N'-dicyclohexylcarbodiimide ( 217 mg), and the mixture was stirred at room temperature for 3 hours. The insoluble solid formed was filtered off and the filtrate was added to (S)-3-[(2S,3R,4R,5S)-2,3,4,6-tetrahydroxy-5-(L-leucyl)aminohexyl Acyl]amino-3-phenylpropionic acid (compound 3) (456 mg) and triethylamine (0.139 ml) in dimethylformamide (100 ml). The mixture was stirred at room temperature for 19 hours, then concentrated under reduced pressure. 1N Hydrochloric acid was added to the residue and the whole was extracted with ethyl acetate-acetonitrile. The organic layer was washed with saturated brine and dried over anhydrous sodium sulfate. The organic solvent was removed and the resulting residue was dissolved in methanol (30 mL) and stirred with 10% palladium/charcoal (200 mg) at room temperature under hydrogen atmosphere for 24 hours. After filtration, the filtrate was concentrated under reduced pressure to obtain a residue which was purified by flash silica gel column chromatography, and then eluted with acetonitrile-water (4:1). Effective fractions were combined and concentrated under reduced pressure, and the resulting residue was passed through a DIAION HP-20SS column (Mitsubishi Kasei Co.), and then eluted with water-acetonitrile. Effective fractions were combined and concentrated under reduced pressure. The residue was recrystallized from methanol-ether to give the title compound (323 mg).

¹ H-NMR (DMSO-d ₆ ) δ: 0.91 (3H, d, J = 6.2 Hz),

0.95(3H, d, J=6.2Hz), 1.50-1.80(3H, m),

2.69(2H, d, J=6.2Hz), 3.20-4.50(10H, m),

5.33 (1H, t, J=6.2Hz), 6.95-7.50 (9H, m),

7.73(1H, d, J=7.4Hz).

Example 88

(S)-3-[(2S,3R,4R,5S)-2,3,4,6-tetrahydroxy-5-(N-(2-aminoisobutyryl)-L-leucyl)aminohexyl Acyl]amino-3-phenylpropionic acid

To a solution of 2-(benzyloxycarbonylamino)isobutyric acid (81 mg) in acetonitrile (10 mL) was added N-hydroxysuccinimide (41 mg) and N,N'-dicyclohexylcarbodiimide (71 mg), the mixture was stirred at room temperature for 3 hours. The insoluble solid formed was filtered off and the filtrate was added to (S)-3-[(2S,3R,4R,5S)-2,3,4,6-tetrahydroxy-5-(L-leucyl)aminohexyl Acyl]amino-3-phenylpropionic acid (compound 3) (150 mg) and triethylamine (0.061 ml) in dimethylformamide (30 ml). The mixture was stirred at room temperature for 22 hours, then concentrated under reduced pressure. The residue was dissolved in methanol (30 mL) and stirred with 10% palladium/charcoal (70 mg) at room temperature under hydrogen atmosphere for 1 day. After filtration, the filtrate was concentrated under reduced pressure, and the obtained residue was passed through a DIAION HP-20SS column (Mitsubishi Kasei Co.), and then eluted with water-acetonitrile. Effective fractions were combined and concentrated under reduced pressure. The residue was recrystallized from methanol-diisopropyl ether to obtain the title compound (114 mg).

¹ H-NMR (D ₂ O) δ: 0.89 (3H, d, J=5.4Hz), 0.94 (3H, d, J=5.8Hz),

1.62(3H, s), 1.64(3H, s), 1.60-1.85(3H, m),

2.74(2H, d, J=7.0Hz), 3.60-3.80(3H, m),

3.90(1H, d, J=9.9Hz), 4.22-4.48(3H, m),

5.21(1H, t, J=7.0Hz), 7.32-7.48(5H, m).

Example 89

(S)-3-[(2S,3R,4R,5S)-5-(N-(1-aminocyclohexyl)carbonyl)-L-leucyl)amino-2,3,4,6-tetrahydroxy Hexanoyl]amino-3-phenylpropanoic acid

The title compound was prepared in the same manner as described in Example 88, substituting 1-(benzyloxycarbonylamino)cyclohexanecarboxylic acid for 2-(benzyloxycarbonylamino)isobutyric acid.

¹ H-NMR (DMSO-d ₆ ) δ: 0.84-0.90 (6H, m), 1.10-1.84 (13H, m),

2.69-2.75(2H, m), 3.15-4.09(10H, m), 4.13(1H, s),

4.38(1H, m), 5.20-5.28(2H, m), 5.76(1H, m), 7.21-

7.40(5H, m), 7.49(1H, m), 8.10-8.28(2H, m).

Example 90

(S)-3-[(2S,3R,4R,5S)-2,3,4,6-tetrahydroxy-5-(L-methionyl-L-leucyl)aminocaproyl]amino-3 -Phenylpropionic acid

To a solution of N-tert-butoxycarbonyl-L-methionine (86 mg) in acetonitrile (10 ml) was added N-hydroxysuccinimide (41 mg) and N,N'-dicyclohexylcarbodiene Amine (71 mg), and the mixture was stirred at room temperature for 2.5 hours. The insoluble solid formed was filtered off and the filtrate was added to (S)-3-[(2S,3R,4R,5S)-2,3,4,6-tetrahydroxy-5-(L-leucyl)aminohexyl Acyl]amino-3-phenylpropionic acid (compound 3) (150 mg) and triethylamine (0.061 ml) in dimethylformamide (30 ml). The mixture was stirred at room temperature for 18.5 hours, then concentrated under reduced pressure. To the residue was added 4N hydrogen chloride in ethyl acetate (5 ml) and the whole was stirred at room temperature for 2 hours. The organic solvent was removed, and the obtained residue was passed through a DIAION HP-20SS column (Mitsubishi Kasei Co.), and then eluted with water-acetonitrile. Effective fractions were combined and concentrated under reduced pressure. The residue was recrystallized from methanol-ethyl acetate to obtain the title compound (60 mg).

¹ H-NMR (DMSO-d ₆ ) δ: 0.84-0.91 (6H, m), 1.45-1.99 (5H, m),

2.03(3H, s), 2.47-2.56(2H, m), 2.69-2.75(2H, m), 3.40-

4.00(6H,m), 4.13(1H,s), 4.37(1H,m), 5.21(1H,m), 7.21-

7.38(5H, m), 7.49(1H, br d, J=8.8Hz), 8.18(1H, m),

8.29(1H, br d, J=7.0Hz).

Example 91

(S)-3-[(2S,3R,4R,5S)-2,3,4,6-tetrahydroxy-5-((S)-methyl-L-cysteinyl)-L-leucine Acyl)aminocaproyl]amino-3-phenylpropionic acid

Following the same procedure described in Example 90, substituting N-tert-butoxycarbonyl-S-methyl-L-cysteine for N-tert-butoxycarbonyl-L-methionine, the title compound was prepared.

¹ H-NMR (DMSO-d ₆ ) δ: 0.84-0.91 (6H, m), 1.46-1.73 (3H, m),

1.91(2H, m), 2.05(3H, s), 2.55-2.88(4H, m), 3.40-

4.09(6H,m), 4.13(1H,s), 4.38(1H,m), 5.22(1H,m), 7.21-

7.38(5H, m), 7.50(1H, br d, J=8.4Hz), 8.14-8.25(2H, m).

Example 92

(S)-3-[(2S,3R,4R,5S)-5-(N-((S)-2-amino-4-pentanoyl)-L-leucyl)amino-2,3,4 , 6-Tetrahydroxyhexanoyl]amino-3-phenylpropanoic acid

Following the same procedure described in Example 90, substituting (S)-2-t-butoxycarbonylamino-4-pentenoic acid for N-t-butoxycarbonyl-L-methionine, the title compound was prepared.

¹ H-NMR (DMSO-d ₆ ) δ: 0.84-0.90 (6H, m), 1.43-1.64 (3H, m),

2.12-2.48(2H, m), 2.69-2.75(2H, m), 3.40-4.09(6H, m),

4.13(1H, s), 4.38(1H, m), 5.00-5.28(3H, m), 5.76(1H, m),

7.21-7.40 (5H, m), 7.49 (1H, br d, J=8.6Hz), 8.13 (1H, br

d, J=7.6Hz), 8.28(1H, br d, J=10.8Hz).

Example 93

(S)-3-[(2S,3R,4R,5S)-2,3,4,6-tetrahydroxy-5-(N-((S)-2-pyrrolidone-5-carbonyl)-L-leu Aminoacyl)aminocaproyl]amino-3-phenylpropionic acid

To a solution of L-pyroglutamic acid (44 mg) in acetonitrile (10 mL) was added N-hydroxysuccinimide (41 mg) and N, N'-dicyclohexylcarbodiimide (71 mg), and the mixture Stir at room temperature for 2 hours. The insoluble solid formed was filtered off and the filtrate was added to (S)-3-[(2S,3R,4R,5S)-2,3,4,6-tetrahydroxy-5-(L-leucyl)aminohexyl Acyl]amino-3-phenylpropionic acid (compound 3) (150 mg) and triethylamine (0.061 ml) in dimethylformamide (30 ml). The mixture was stirred at room temperature for 22 hours, then concentrated under reduced pressure. The residue was passed through a DIAION HP-20SS column (Mitsubishi Kasei Co.), and then eluted with water-acetonitrile. Effective fractions were combined and concentrated under reduced pressure. The residue was passed through a Sephadex LH-20 column (Pharmacia, Sweden) and eluted with water. Effective fractions were combined and concentrated under reduced pressure. The residue was recrystallized from methanol-ethyl acetate to obtain the title compound (24 mg).

¹ H-NMR (CD ₃ OD) δ: 0.94 (3H, d, J = 6.2 Hz),

0.97(3H, d, J=6.6Hz), 1.62-1.78(3H, m), 2.11-2.48(4H, m),

2.89(2H, t, J=6.4Hz), 3.65-3.72(3H, m),

3.91(1H, d, J=9.8Hz), 4.19-4.26(2H, m),

4.32(1H, d, J=1.4Hz), 4.50(1H, m), 5.38(1H, t, J=6.4Hz),

7.23-7.42(5H, m).

Example 94

(S)-3-[(2S,3R,4R,5S)-2,3,4,6-tetrahydroxy-5-((N,N-dimethyl-L-valyl)-L-leu Aminoacyl)aminocaproyl]amino-3-phenylpropanoic acid diphenylmethyl ester

To N,N-dimethyl-L-valine (58 mg) and (S)-3-[(2S,3R,4R,5S)-2,3,4,6-tetrahydroxy-5-( Diethyl cyanophosphonate (82 mg ) and triethylamine (0.14 ml), the mixture was stirred at room temperature for 18 hours. After adding water, the whole was extracted with ethyl acetate, and the extract was washed with saturated brine and dried over anhydrous sodium sulfate. The residue obtained by removing the organic solvent was recrystallized from ethyl acetate-hexane to obtain the title compound (282 mg).

¹ H-NMR (DMSO-d ₆ ) δ: 0.70-0.95 (12H, m), 1.00-2.00 (4H, m),

2.17(6H, s), 3.04(2H, d, J=7.0Hz), 3.30-5.40(12H, m),

6.67(1H, s), 7.10-7.40(16H, m), 8.01(1H, d, J=8.8Hz),

8.15(1H, d, J=8.8Hz).

Example 95

(S)-3-[(2S,3R,4R,5S)-2,3,4,6-tetrahydroxy-5-(L-valyl-(N-methyl-L-leucyl)amino Hexanoyl]amino-3-phenylpropanoic acid

To a solution of (N-tert-butoxycarbonyl-L-valyl)-N-methyl-L-leucine (240 mg) in acetonitrile (20 mL) was added N-hydroxysuccinimide (88 mg ) and N, N'-dicyclohexylcarbodiimide (151 mg), and the mixture was stirred at room temperature for 3 hours. The insoluble solid formed was filtered off and the filtrate was added to (S)-3-[(2S,3R,4R,5S)-5-amino-2,3,4,6-tetrahydroxyhexanoyl]amino-3-benzene Propionic acid (compound 5) (239 mg) and triethylamine (0.097 ml) in dimethylformamide (70 ml). The mixture was stirred at room temperature for 16 hours, then concentrated under reduced pressure. To the residue was added 4N hydrogen chloride in ethyl acetate (20 ml) and the whole was stirred at room temperature for 1 hour. The organic solvent was removed, and the obtained residue was passed through a DIAION HP-20SS column (Mitsubishi Kasei Co.), and then eluted with water-acetonitrile. Effective fractions were combined and concentrated under reduced pressure. The residue was recrystallized from methanol-ether to obtain the title compound (123 mg).

¹ H-NMR (DMSO-d ₆ ) δ: 0.70-1.00 (12H, m), 1.00-2.20 (4H, m),

2.50-2.80(2H, m), 2.86(3/2H, s), 2.89(3/2H, s), 3.00-

5.30(9H, m), 7.10-7.40(5H, m).

Example 96

(S)-3-[(2S,3R,4R,5S)-5-(N-((S)-2-amino-3-methylbutyl)-L-leucyl)amino-2,3 , 4,6-Tetrahydroxyhexanoyl]amino-3-phenylpropanoic acid

At 0°C, N-tert-butoxycarbonyl-L-valine (240 mg) and (S)-3-[(2S,3R,4R,5S)-2,3,4,6-tetrahydroxy Sodium cyanoborohydride (63 mg) was added to a solution of diphenylmethyl 5-(L-leucyl)aminocaproyl]amino-3-phenylpropanoate (278 mg) in methanol (10 ml), The mixture was stirred at 0°C for 2 hours and at room temperature for 18 hours. After concentration under reduced pressure, the residue was purified by flash silica gel column chromatography, and then eluted with ethyl acetate-methanol (20:1). Effective fractions were combined and concentrated under reduced pressure. To the residue was added 4N hydrogen chloride in ethyl acetate (10 ml) and the whole was stirred at room temperature for 1 hour. The organic solvent was removed, and the obtained residue was passed through a DIAION HP-20SS column (Mitsubishi Kasei Co.), and then eluted with water-acetonitrile. Effective fractions were combined and concentrated under reduced pressure. The residue was recrystallized from methanol-ether to give the title compound (25 mg).

¹ H-NMR (DMSO-d ₆ +3% TFA) δ: 0.85-1.10 (6H, m), 1.40-

2.60(4H, m), 2.70-2.90(2H, m), 2.90-5.40(9H, m), 7.20-

7.45(5H, m), 8.00-8.40(2H, m).

Example 97

(S)-3-[(2S,3R,4R,5S)-2,3,4,6-tetrahydroxy-5-((S)-2-(L-norvalyl)amino-4-pentane Enyl)aminocaproyl]amino-3-phenylpropanoic acid

To a solution of N-tert-butoxycarbonyl-L-norvaline (54 mg) in acetonitrile (1 mL) was added N-hydroxysuccinimide (29 mg) and N,N'-dicyclohexylcarbodi imine (52 mg), and the mixture was stirred at room temperature for 3 hours. The insoluble solid formed was filtered off, and the filtrate was concentrated under reduced pressure. The residue was dissolved in dimethylformamide (8 mL). (S)-3-[(2S,3R,4R,5S)-5-((S)-2-amino-4-pentenoyl)amino-2,3,4,6-tetrahydroxyhexyl Acyl]-3-phenylpropanoic acid (110 mg) and triethylamine (0.035 ml). The mixture was stirred at room temperature for 20 hours, then concentrated under reduced pressure. To the residue was added 4N hydrogen chloride in ethyl acetate (10 ml) and the whole was stirred at room temperature for 1 hour. The organic solvent was removed, and the obtained residue was passed through a DIAION HP-20SS column (Mitsubishi Kasei Co.), and then eluted with water-acetonitrile. Effective fractions were combined and concentrated under reduced pressure. The residue was recrystallized from methanol-ether to obtain the title compound (56 mg).

¹ H-NMR (DMSO-d ₆ ) δ: 0.86 (3H, t, J=7.0Hz), 1.20-

1.80 (4H, m), 2.20-2.90 (4H, m), 3.30-5.90 (12H, m), 7.10-

7.45(5H, m), 7.54(1H, d, J=7.8Hz), 8.28(1H, d, J=12.8Hz).

Example 98

(S)-3-[(2S,3R,4R,5S)-2,3,4,6-tetrahydroxy-5-((S)-2-(L-isoleucyl)amino-4-pentane Enyl)aminocaproyl]amino-3-phenylpropanoic acid

Following the same procedure described in Example 97, substituting N-tert-butoxycarbonyl-L-isoleucine for N-tert-butoxycarbonyl-L-norvaline, the title compound was prepared.

¹ H-NMR (DMSO-d ₆ ) δ: 0.75-0.95 (6H, m), 0.95-1.80 (3H, m),

2.10-2.90(4H,m), 3.10-4.50(8H,m), 4.95-5.90(4H,m),

7.15-7.20 (5H, m).

Example 99

(S)-3-[(2S,3R,4R,5S)-2,3,4,6-tetrahydroxy-5-((S)-2-(L-methionyl)amino-4-pentane Enyl)aminocaproyl]amino-3-phenylpropanoic acid

Following the same procedure as described in Example 97, substituting N-tert-butoxycarbonyl-L-methionine for N-tert-butoxycarbonyl-L-norvaline, the title compound was prepared.

¹ H-NMR (DMSO-d ₆ ) δ: 1.50-2.10 (2H, m), 2.10-3.00 (6H, m),

2.03(3H, s), 3.20-5.90(9H, m), 7.15-7.40(5H, m).

Example 100

(S)-3-[(2S,3R,4R,5S)-2,3,4,6-tetraacetoxy-5-(N-benzyloxycarbonyl-L-leucyl)aminocaproyl]amino -Ethyl 3-(4-methylphenyl)propionate

To [(2S,3R,4R,5S)-2,3,4,6-tetraacetoxy-5-(N-benzyloxycarbonyl-L-leucyl)aminocaproic acid (250 mg) in acetonitrile ( 10 ml) solution was added N-hydroxyl-5-norbornene-2,3-dicarboxyimide (110 mg) and N,N'-dicyclohexylcarbodiimide (110 mg), the mixture was at room temperature Stir for 1 hour. After adding ethyl (S)-3-amino-3-(4-methylphenyl)propionate (150 mg) and triethylamine (0.13 ml) in acetonitrile (10 ml), the whole was stirred at room temperature for 18 Hour. The insoluble solid formed was filtered off, and the filtrate was concentrated under reduced pressure. The residue was dissolved in ethyl acetate (100 mL) and washed with saturated brine (50 mL×2), then dried over anhydrous sodium sulfate. After concentration under reduced pressure, the residue was purified by silica gel column chromatography, and then eluted with ethyl acetate-hexane (2:1). Effective fractions were combined and concentrated under reduced pressure to obtain the title compound (184 mg).

¹ H-NMR (CD ₃ OD) δ: 0.89-0.96 (6H, m), 1.15 (3H, q, J=7.2Hz),

1.45-1.73(3H, m), 1.96(3H, s), 2.01(3H, s), 2.04(3H, s),

2.08(3H, s), 2.28(3H, s), 2.76(1H, dd, J=15.8Hz, 7.4Hz),

2.88(1H, dd, J=15.8Hz, 7.4Hz), 3.80(1H,dd, J=11.0Hz, 7.2Hz),

3.99-4.21(7H, m), 4.51(1H, t, J=7.4Hz), 5.07(2H, s),

5.22 (1H, t, J = 7.4Hz), 5.37 (2H, s), 7.10 (2H, d, J = 8.0Hz),

7.18(2H, d, J=8.0Hz), 7.30-7.36(5H, m).

Example 101

(S)-3-[(2S,3R,4R,5S)-2,3,4,6-tetrahydroxy-5-(L-leucyl)aminocaproyl]amino-3-(4-methyl Phenyl)propionic acid

(S)-3-[(2S,3R,4R,5S)-2,3,4,6-tetraacetoxy-5-(N-benzyloxycarbonyl-L-leucyl)aminocaproyl] A solution of ethyl amino-3-(4-methylphenyl)propanoate (184 mg) in methanol (10 mL) was stirred with 10% palladium/charcoal (100 mg) at room temperature under an atmosphere of hydrogen for 2 hours. After filtration, the filtrate was concentrated under reduced pressure. The residue was dissolved in methanol (10 ml), and to the solution was added 1N aqueous sodium hydroxide solution (2.2 ml) under ice-cooling. The mixture was stirred under ice-cooling for 1 hr, and then 1N hydrochloric acid (2.2 ml) was added. After concentration under reduced pressure, the residue was passed through a DIAION HP-20SS column (Mitsubishi Kasei Co.), and then eluted with water-acetonitrile. Effective fractions were combined and concentrated under reduced pressure. The residue was recrystallized from methanol-ethyl acetate to obtain the title compound (50 mg).

¹ H-NMR (CD ₃ OD) δ: 0.98-1.02 (6H, m), 1.60-1.85 (3H, m),

2.27(3H, s), 2.67(2H, d, J=6.4Hz), 3.65-3.74(3H, m), 3.85-

3.90(2H, m), 4.24-4.31(2H, m), 5.27(1H, t, J=6.4Hz),

7.09(2H, d, J=8.0Hz), 7.26(2H, d, J=8.0Hz).

The structural formulas of the compounds obtained in Reference Examples and Examples are shown below. The abbreviation 'Ac' means acetyl.

Test example 1

In vitro antibacterial test: antibacterial activity against Helicobacter pylori in vitro

Using Helicobacter pylori (NCTC 11637) as a test strain, the antibacterial activities of HC-70I (compound 1) and HC-70II (compound 2) were determined by the following agar dilution method. HC-70I and HC-70II were separately dissolved in dimethyl sulfoxide, and using sterile distilled water, a two-fold dilution series for use as samples was prepared. Using 7% horse blood-supplemented Brucella agar as the medium, plates were prepared by mixing 2 ml of each sample with 18 ml of 7% horse blood-brucella agar. To prepare the inoculum, Helicobacter pylori was cultured with shaking in 2.5% fetal bovine serum-brucella broth at 37°C for 20 days using a gas pak container filled with CampyPak ^TM [BBL Beckton Dickinson Microbiology Systems]. Hour. Assay plates were inoculated with 5 microliters of each cell suspension adjusted to ¹⁰⁶ CFU/ml with 2.5% FBS-Brucella Broth and placed in an inflated container containing a CampyPak ^™ and a water-soaked tampon , kept at 37°C for 4 days. After incubation, bacterial growth was roughly assessed and the lowest concentration at which no growth was observed was recorded as the MIC (Minimum Inhibitory Concentration). For HC-70I and HC-70II, the MIC value is 0.025 (microgram/ml).

Test example 2

In vivo antibacterial test:

Mice (Crj: ICR, male, 5 weeks old) were fasted for 20 hours and each mouse was inoculated with Helicobacter pylori TN2F4 at a dose of 10 ^7.79 CFU. From 11 days after the infection, 50 mg/kg of the test compound suspended in 0.5% methylcellulose/water was orally administered twice a day in the morning and evening for two consecutive days. On the day after the last dose, stomachs were isolated from infected mice and homogenized, and a 10-fold dilution series of the homogenates were inoculated on charcoal-modified Skirrow medium. Cultivate at 37°C for 4 days under the condition of micro-aeration, and determine the clearance rate according to the growth of bacteria.

The results are shown in Table 1. Bacteria counts are expressed as mean ± standard error, and statistical analysis was performed by comparison with the control group according to Dunnett's method.

Table 1 sample Dose (mg/kg) clearance rate(%) Bacterial recovery (LogCFU/gastric wall) Control (0.5% methylcellulose) 0 0/4(0) 4.67±0.06 HC-70II.HCl 50 4/4(100) ND HC-70III 50 4/4(100) ND

ND: not found

As can be seen from Table 1, HC-70II hydrochloride (compound 4) and HC-70III (compound 3) achieved 100% clearance at the 50 mg/kg dose level. Therefore, it is apparent that the pharmaceutical composition of the present invention is effective in the prevention and treatment of gastritis, gastric ulcer, duodenal ulcer and gastric cancer associated with Helicobacter pylori.

Test example 3

5-week-old MON/Jms/Gbs Mongolian gerbils were intragastrically inoculated with 10 ^7.58 CFU of Helicobacter pylori TN2GF4. Four weeks after infection, the compound of Example 47 suspended in 0.5% methylcellulose was orally administered twice a day at a dose of 30 mg/kg for two consecutive days. The day after the final treatment, the animals were sacrificed.

The stomachs were removed and homogenized with 3 ml of Brucella broth, and the number of bacteria in the homogenate was determined by serial dilution and titration on modified Skirrow plates. Plates were incubated at 37°C for 4 days under microaerobic conditions prior to counting. The absence of H. pylori in the stomach on the day after the final treatment was defined as complete eradication.

The compound of Example 47 at a dose of 30 mg/kg twice daily for two days reduced the number of infected organisms; 2 out of 4 gerbils achieved complete clearance.

Table 2 The inhibitory effect of repeated administration of the compound of Example 47 on gastric infection caused by Helicobacter pylori TN2GF4 in MON/Jms/Gbs Mongolian gerbils compound Dose (mg/kg) Clearance Rate Cleared / Total (%) Bacterial recovery Log CFU/gastric wall mean ± SE Vehicle control 0 0/3(0) 6.05±0.08 Compound of Example 47 30 2/4(50) 2.23±0.44 ^**

^** According to Dunnett's test, p∠0.01 for vehicle control

Test example 4

Anti-Helicobacter Pylori Effect of Gastric Mucoadhesive Preparations in Vivo

Mongolian gerbils (MON/Jms/Gbs) infected with Helicobacter pylori were respectively with 3 mg/kg, 10 mg/kg HC-70-II twice a day for 7 consecutive days of oral preparation containing HC-70 obtained in Example 3 -II gastric mucoadhesive preparation (HC-70-II AdMMS-1 in Table 3), and 0.5% methylcellulose suspension containing HC-70-II (HC-70-II suspension in Table 3 ). Sixteen hours after the final dose, the stomach was removed, the stomach wall was homogenized, and serial dilutions were plated on H. pylori selective medium. The inoculated medium was incubated at 37°C for 4 days under minimal oxygen aeration and the number of viable cells was counted. The results are shown in Table 3.

table 3 formula Dose (mg/kg) HC-70-II Bacterial recovery Log CFU/gastric wall mean ± SE control 0 6.69±0.19 HC-70-II AdMMs-1 3 4.11±1.08 HC-700-II, suspension 10 4.09±0.80

Compared with HC-70-II suspension, the gastric mucoadhesive formulation containing HC-70-II had the same level of anti-inflammatory activity as HC-70-II suspension (which was 1/3 dose of HC-70-II suspension). Helicobacter pylori activity.

Preparation example 1

As a therapeutic agent for Helicobacter pylori infection, the compound or salt of the present invention is generally administered in the following dosage forms.

1. Capsules

(1) HC-70I 100 mg

(2) Lactose 90 mg

(3) Microcrystalline Cellulose 70 mg

(4) Magnesium stearate 10 mg

270 mg/capsule

Mix and granulate (1), (2), and whole amounts of (3) and half of (4). The remaining (4) is added to the granulation and the whole composition is filled into the gelatin capsule wall.

2. Tablets

(1) HC-70I 100 mg

(2) Lactose 35 mg

(3) Corn starch 150 mg

(4) Microcrystalline Cellulose 30 mg

(5) Magnesium stearate 5 mg

Each tablet weighs 320 mg

Mix and granulate (1), (2), and whole amounts of (3), 2/3 of (4) and half of (5). The remaining (4) and (5) are added to the granulation and the whole composition is compressed into tablets.

Preparation example 2

A mixture of hydrogenated castor oil (Lubri Wax 101 ^™ , Freund Industries Co., Ltd.) (40 g) and hexa(tetra)glycerol behenate (HB-310 ^™ , Sakamoto Yakuhin Kogyo Co., Ltd.) (39 g) was carried out at 85° C. melt. Add 1 gram of compound 2 (HC-70-II), 10 grams of acrylic polymer (HIVISWAKO 104 ^™ , Wako Pure Chemical Industry Co., Ltd.) and 10 grams of low-substituted hydroxypropyl cellulose (L -HPC ^™ , Shin-Etsu Chemicals), the mixture was stirred and dispersed at a constant temperature of 85° C. for 2 hours. This molten mixture was dropped into a 15 cm (diameter) aluminum pan rotating at 2700 rpm at a flow rate of 50 g/min to obtain spherical fine particles passing through 42 mesh.

Preparation example 3

A mixture of hydrogenated castor oil (Lubri Wax 101 ^™ , Freund Industries Co., Ltd.) (20 g) and hexa(tetra)glyceride behenate (HB-310 ^™ , Sakamoto Yakuhin Kogyo Co., Ltd.) (59 g) was carried out at 85° C. melt. Add 1 gram of compound 2 (HC-70-II), 10 grams of acrylic polymer (HIVISWAKO 104 ^™ , Wako Pure Chemical Industry Co., Ltd.) and 10 grams of low-substituted hydroxypropyl cellulose (L -HPC ^™ , Shin-Etsu Chemicals), the mixture was stirred and dispersed at a constant temperature of 85° C. for 2 hours. This molten mixture was dropped into a 15 cm (diameter) aluminum pan rotating at 2700 rpm at a flow rate of 50 g/min to obtain spherical fine particles passing through 42 mesh.

Preparation example 4

A mixture of hexa(tetra)glycerol behenate (HB-310 ^™ , Sakamoto Yakuhin Kogyo Co., Ltd.) (69 g) was melted at 80°C. Add 1 gram of compound 2 (HC-70-II), 10 grams of acrylic polymer (HIVISWAKO 104 ^™ , Wako Pure Chemical Industry Co., Ltd.) and 20 grams of low-substituted hydroxypropyl cellulose (L -HPC ^™ , Shin-Etsu Chemicals), the mixture was stirred and dispersed at a constant temperature of 80° C. for 2 hours. This molten mixture was dropped into a 15 cm (diameter) aluminum pan rotating at 2400 rpm at a flow rate of 50 g/min to obtain spherical fine particles passing through 42 mesh.

Preparation example 5

A mixture of hydrogenated castor oil (Lubri Wax 101 ^™ , Freund Industries Co., Ltd.) (30 g) and hexa(tetra)glyceride behenate (HB-310 ^™ , Sakamoto Yakuhin Kogyo Co., Ltd.) (49 g) was carried out at 85° C. melt. Add 1 gram of compound 2 (HC-70-II), 10 grams of acrylic polymer (HIVISWAKO 104 ^™ , Wako Pure Chemical Industry Co., Ltd.) and 10 grams of low-substituted hydroxypropyl cellulose (L -HPC ^™ , Shin-Etsu Chemicals), the mixture was stirred and dispersed at a constant temperature of 85° C. for 2 hours. This molten mixture was dropped into a 15 cm (diameter) aluminum pan rotating at 2700 rpm at a flow rate of 50 g/min to obtain spherical fine particles passing through 42 mesh.

Preparation example 6

A mixture of hydrogenated castor oil (Lubri Wax 101 ^™ , Freund Industries Co., Ltd.) (20 g) and hexa(tetra)glyceride behenate (HB-310 ^™ , Sakamoto Yakuhin Kogyo Co., Ltd.) (59 g) was carried out at 85° C. melt. Add 1 gram of compound 2 (HC-70-II), 10.0 grams of acrylic polymer (FX-214 ^™ , BF Goodrich Industries, Ltd.) and 10 grams of low-substituted hydroxypropyl cellulose (L -HPC ^™ , Shin-Etsu Chemicals), the mixture was stirred and dispersed at a constant temperature of 85° C. for 2 hours. This molten mixture was dropped into a 15 cm (diameter) aluminum pan rotating at 2700 rpm at a flow rate of 50 g/min to obtain spherical fine particles passing through 42 mesh.

Example 7

A mixture of hexa(tetra)glycerol behenate (HB-310 ^™ , Sakamoto Yakuhin Kogyo Co., Ltd.) (60 g) was melted at 80°C. Add 30 grams of compound 2 (HC-70-II), 6 grams of acrylic polymer (HIVISWAKO 104 ^™ , Wako Pure Chemical Industry Co., Ltd.) and 4 grams of low-substituted hydroxypropyl cellulose (L -HPC ^™ , Shin-Etsu Chemicals), the mixture was stirred and dispersed at a constant temperature of 80° C. for 2 hours. This molten mixture was dropped into a 15 cm (diameter) aluminum pan rotating at 3960 rpm at a flow rate of 50 g/min to obtain spherical fine particles passing through 42 mesh.

Preparation example 8

A mixture of hydrogenated castor oil (Lubri Wax 101 ^™ , Freund Industries Co., Ltd.) (10 g) and hexa(tetra)glycerol behenate (HB-310 ^™ , Sakamoto Yakuhin Kogyo Co., Ltd.) (50 g) was carried out at 85° C. melt. Add 30 grams of compound 2 (HC-70-II), 6 grams of acrylic polymer (HIVISWAKO 104 ^™ , WakoPure Chemical Industry Co., Ltd.) and 4 grams of low-substituted hydroxypropyl cellulose (L- HPC ^™ , Shin-Etsu Chemicals), the mixture was stirred and dispersed at 85° C. for 2 hours. This molten mixture was dropped into a 15 cm (diameter) aluminum pan rotating at 3960 rpm at a flow rate of 50 g/min to obtain spherical fine particles passing through 42 mesh.

Test example 9

A mixture of hexa(tetra)glycerol behenate (HB-310 ^™ , Sakamoto Yakuhin Kogyo Co., Ltd.) (60 g) was melted at 80°C. Add 30 grams of compound 2 (HC-70-II), 6 grams of acrylic polymer (EX-214 ^™ , BFGoodrich Industries, Ltd.) and 4 grams of low-substituted hydroxypropyl cellulose (L -HPC ^™ , Shin-Etsu Chemicals), the mixture was stirred and dispersed at a constant temperature of 80° C. for 2 hours. This molten mixture was dropped into a 15 cm (diameter) aluminum pan rotating at 3960 rpm at a flow rate of 50 g/min to obtain spherical fine particles passing through 42 mesh.

Preparation example 10

A mixture of hydrogenated castor oil (Lubri Wax 101 ^™ , Freund Industries Co., Ltd.) (10 g) and hexa(tetra)glycerol behenate (HB-310 ^™ , Sakamoto Yakuhin Kogyo Co., Ltd.) (50 g) was carried out at 85° C. melt. Add 30 grams of compound 2 (HC-70-II), 6 grams of acrylic polymer (EX-214 ^™ , BF Goodrich Industries, Ltd.) and 4 grams of low-substituted hydroxypropyl cellulose (L -HPC ^™ , Shin-Etsu Chemicals), the mixture was stirred and dispersed at a constant temperature of 85°C for 2 hours. This molten mixture was dropped into a 15 cm (diameter) aluminum pan rotating at 3960 rpm at a flow rate of 50 g/min to obtain spherical fine particles passing through 42/119 mesh.

Industrial applicability

The compound (I) of the present invention has outstanding and very high antibacterial activity against Helicobacter pylori bacteria represented by Helicobacter pylori. Therefore, the compound (I) can achieve the desired anti-H. pylori effect at significantly lower doses compared to conventional antibacterial agents used to control Helicobacter pylori bacteria, especially Helicobacter pylori.

Compound (I) is effective in preventing and treating various diseases related to Helicobacter pylori, such as duodenal ulcer, gastric ulcer, chronic gastritis and gastric cancer. In addition, since Helicobacter pylori is the main factor of ulcer recurrence, compound (I) is also effective in preventing ulcer recurrence.

In addition, compound (I) is to some Gram-positive bacteria such as Staphylococcus vulgaris and Bacillus and some Gram-negative bacteria such as Escherichia vulgaris, Pseudomonas, Proteus, Klebsiella, Serratia Bacteria, Salmonella, Citrobacter, Alcalibacteria, etc. are inactive. Therefore, compound (I) can selectively prevent or treat diseases related to Helicobacter pylori, but has weak effects on other bacteria and fungi, so it can be used as a safe drug.

The gastric mucoadhesive composition of the present invention can reduce the amount of the active ingredient to half to one-twentieth of the dosage of its suspension.

Claims (38)

1. formula (I) compound

R wherein ¹Represent commutable amino; R ²But representative esterification or amidated carboxyl; R ³, R ⁴, R ⁵, and R ⁶Represent the hydroxyl that to protect respectively; Q represents commutable aryl; Or its salt.

2. according to the compound of claim 1, R wherein ¹Be amido or the amino that replaced by commutable alkyl.

3. according to the compound of claim 2, the amino that replaced by amino-acid residue of amido wherein.

4. according to the compound of claim 3, wherein amino-acid residue is the a-amino acid residue.

5. according to the compound of claim 1, R wherein ¹Be amino or following formula group:

R wherein ⁷Be the amino that can be replaced by α-L-amino-acid residue that α-L-amino-acid residue replaces, R ⁸It is commutable alkyl; R ²Representation carboxy; R ³, R ⁴, R ⁵, and R ⁶The difference representation hydroxy; Q represents phenyl.

6. according to the compound of claim 5, it is represented by following formula V:

R wherein ⁷And R ⁸Identical definition with claim 5.

7. according to the compound of claim 5, R wherein ⁸Be C _1-10Alkyl, C _6-14Aryl-C _1-6Alkyl, C _2-10Alkenyl or C _2-10Alkynyl, these groups can be substituted.

8. according to the compound of claim 7, R wherein ⁸Be C _1-6Alkyl or C _2-6Alkenyl.

9. according to the compound of claim 7, R wherein ⁷Be can be by amino valyl, that valyl is valyl, valyl isoleucyl or valyl leucyl replace.

10. according to the compound of claim 8, R wherein ⁸Be isobutyl-or allyl group.

11. according to the compound of claim 1, wherein R ¹Be amino.

12. according to the compound of claim 1, it is (S)-3-[(2S, 3R, and 4R, 5S)-2,3,4, and the amino hexanoyl of 6-tetrahydroxy-5-(L-valyl-L-valyl-L-leucyl)] amino-3-phenylpropionic acid.

13. according to the compound of claim 1, it is (S)-3-[(2S, 3R, and 4R, 5S)-2,3,4, and the amino hexanoyl of 6-tetrahydroxy-5-(L-valyl-L-isoleucyl--L-leucyl)] amino-3-phenylpropionic acid.

14. according to the compound of claim 1, it is (S)-3-[(2S, 3R, and 4R, 5S)-2,3,4, and the amino hexanoyl of 6-tetrahydroxy-5-(L-valyl-L-leucyl-L-leucyl)] amino-3-phenylpropionic acid.

15. according to the compound of claim 1, it is (S)-3-[(2S, 3R, and 4R, 5S)-2,3,4, and the amino hexanoyl of 6-tetrahydroxy-5-(L-valyl-L-leucyl)] amino-3-phenylpropionic acid.

16. according to the compound of claim 1, it is (S)-3-[(2S, 3R, and 4R, 5S)-2,3,4, and the amino hexanoyl of 6-tetrahydroxy-5-(L-leucyl)] amino-3-phenylpropionic acid.

17. according to the compound of claim 1, it is (S)-3-[(2S, 3R, and 4R, 5S)-and 5-((S)-2-amino-4-amylene acyl) amino-2,3,4,6-tetrahydroxy hexanoyl] amino-3-phenylpropionic acid.

18. according to the compound of claim 1, it is (S)-3-[(2S, 3R, and 4R, 5S)-and 5-(the amino butyryl of (S)-2-) amino-2,3,4,6-tetrahydroxy hexanoyl] amino-3-phenylpropionic acid.

19. according to the compound of claim 1, it is (S)-3-[(2S, 3R, and 4R, 5S)-2,3,4, and the amino hexanoyl of 6-tetrahydroxy-5-(L-isoleucyl--L-leucyl)] amino-3-phenylpropionic acid.

20. according to the compound of claim 1, it is (S)-3-[(2S, 3R, and 4R, 5S)-2,3,4, and the amino hexanoyl of 6-tetrahydroxy-5-(L-methionyl-L-leucyl)] amino-3-phenylpropionic acid.

21. according to the compound of claim 1, it is (S)-3-[(2S, 3R, and 4R, 5S)-2,3,4,6-tetrahydroxy-5-((S)-2-(L-norvalyl) amino-4-amylene acyl] amino-3-phenylpropionic acid.

22. contain pharmaceutical composition according to the compound of claim 1.

23. according to the composition of claim 22, it is an anti-helicobacter pylori agent.

24. according to the anti-helicobacter pylori agent of claim 23, it is prevention and the curative that is used for the helicobacter pylori infection diseases associated.

25., be stomach ulcer or duodenal ulcer, gastritis, cancer of the stomach or gastric MALT lymphoma wherein with the helicobacter pylori infection diseases associated according to the anti-helicobacter pylori agent of claim 24.

26. anti-helicobacter pylori agent, it comprises according to the compound of claim 1 and at least a other antiseptic-germicide or/and anti-ulcerative drug.

27. according to the composition of claim 22, it is a stomach mucous membrane adhesion pharmaceutical composition.

28. according to the composition of claim 27, it comprises (a) compound according to claim 1, (b) lipid and/or poly-glycerine fatty acid ester and (c) can become the sticky agent of viscosity with water.

29. according to the composition of claim 28, wherein (c) sticky agent is acrylic acid polymer or its salt.

30. according to the composition of claim 28, it comprises the material of this sticky agent of (d) a kind of energy swelling.

31. according to the composition of claim 30, material that wherein can this sticky agent of swelling is curdlan and/or the low hydroxypropylcellulose that replaces.

32. a method for preparing according to the compound of claim 1, it comprises the carboxylic acid that makes formula (II)

R wherein ¹Be commutable amino; R ³, R ⁴, R ⁵, and R ⁶Represent the hydroxyl that can protect respectively, or its salt, or its reactive derivative; With formula (III) compound

R wherein ²But representative esterification or amidated carboxyl; Q represents commutable aryl, or its salt reacts.

33. a method for preparing according to the compound of claim 1, it comprises makes formula (IV) compound R wherein ²But representative esterification or amidated carboxyl; R ³, R ⁴, R ⁵And R ⁶Represent the hydroxyl that can protect respectively, Q represents commutable aryl, or its salt, or its reactive derivative; With following formula: compound:

R ⁹-X is R wherein ⁹Represent acyl group, or commutable alkyl; X represents leavings group, or its salt, or its activity derivatives reaction.

34. method for preparing according to the compound of claim 5, it is included in the substratum growth can produce microorganism strains according to the bacillus (Bacillus) of the compound of claim 5, allows this bacterial strain grow in fermented liquid and accumulates this compound and collection compound.

35. according to the method for claim 34, wherein microorganism strains is genus bacillus HC-70 or unusual genus bacillus HC-72 (Bacillus insolitus HC-72).

36. can produce genus bacillus HC-70 or unusual genus bacillus HC-72 according to the compound of claim 5.

37. with the method for helicobacter pylori infection diseases associated, it comprises formula (I) compound to the administration significant quantity of needs in prevention or the treatment Mammals R wherein ¹Represent commutable amino; R ²But representative esterification or amidated carboxyl; R ³, R ⁴, R ⁵, and R ⁶Represent the hydroxyl that to protect respectively; Q represents commutable aryl; Or its salt.

38. formula (I) compound is used to prepare the purposes of anti-helicobacter pylori agent

R wherein ¹Represent commutable amino; R ²But representative esterification or amidated carboxyl; R ³, R ⁴, R ⁵, and R ⁶Represent the hydroxyl that to protect respectively; Q represents commutable aryl; Or its salt.

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