CA1249400A - Muramyldipeptide derivatives - Google Patents
Muramyldipeptide derivativesInfo
- Publication number
- CA1249400A CA1249400A CA000432987A CA432987A CA1249400A CA 1249400 A CA1249400 A CA 1249400A CA 000432987 A CA000432987 A CA 000432987A CA 432987 A CA432987 A CA 432987A CA 1249400 A CA1249400 A CA 1249400A
- Authority
- CA
- Canada
- Prior art keywords
- carbon atoms
- group
- alanyl
- deoxy
- thio
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired
Links
Abstract
ABSTRACT OF THE DISCLOSURE
Muramyldipeptide derivatives represented by the formula:
Muramyldipeptide derivatives represented by the formula:
Description
MURAMYLDIPEPTIDE DERIVATIVES
FIELD OF THE INVENTION
This invention relates to novel muramyldipeptide derivatives having excellent immunoadjuvant activity and prophylactic and therapeutic effects against microbial infections and antitumor activity and, more speclfically, this invention relates to a muramyldipeptide derivative represented by the formula ~
~ O~SR3 (I) R2~
NH--A c~ 1 CONH
C~3C~ICO-Ala-NH-C~CH2CH~COOR4 . I
wherein Ala represents alanine; Acyl represents an acyl group having 2 to 40 carbon atoms; ~1 and R2 each represents a hydrogen atom, or Rl and R2, when taken together, may : form an alkylidene group having 1 to 6 carbon atoms; R3 represents a hy~rogen atom, an acyl group having 2 to 40 carbon atoms or an alkyl group having 1 to 40 carbon atoms;
and R4 represents a hydrogen atom or an alkyl gxoup having : 1 to 20 carbon atoms.
^' ~
3~
BACKGROUND OF THE INVENTION
Hitherto, certain types of muramyldipeptide derivatives have been known to have useful immunotherapeutic activities. For example, U.S. Paten-t 4,101,536 discloses muramyldipeptide derivatives and the salts thereof having potent immunoadjuvant activi-ty and antltumor activity and being applicable as an agent for the immunotherapy of cancer for human and animals~ These known compaunds, however, have a free hydroxy group at the l-position of muramic acid of M-acetylmuramyldipeptide and do not possess an S-substituted moiety at the.l-position.
. SIJMMARY O`F THE INVENTION
As a result of extensive studies on compounds useful as adjuvant substances, we have found that the compounds of the above formula (I) which are obtained by introducing a sul~ur atom to ~he l-position of muramic acid o N-acylmuramyldipeptides, the minimum unit consti-tuting bact~rial cell wall of human type Mycobacterium tubercul_sis, BCG and other mycobacteria as well as cellular parastic bacteria, have excellent immunoadjuvant activity, and thus completed the present invention.
The compounds according to the present invention are of low toxicity and possess excellent immunoadjuvant activity and, therefore, can be useful for various diseases attributed to reduction in i~munity function~ for example, ., , O~
as prophylactic and therapeutic ayents against microbial infectlons, antitumor agents and the like.
DETAILED DESCRIPTION OF THE INVENTION
The muramyldipeptide derivatives of the present s invention are represented by the formula (I) ~ 9R3 R20 \~ ~
~ ~H-Acyl fONH2 CH3CHCO-Ala-NEI-CHCH2CH2COOR~
wherein Ala represents alanine; Acyl represents an acyl group having 2 to 40 carbon atoms; Rl and R2 each represents a hydrogen atom, or Rl and R2, when taken together, may : 10 form an alkylidene group having 1 to 6 carbon atoms; R3 represents a hydrogen atom, an acyl group having 2 to 40 carbon atoms or an alkyl group having 1 to 40 carbon atoms;
and R4 represents a hydrogen atom or an alkyl group having 1 to 20 carbon atoms.
lS A preferred class of compounds according to the present invention is that having the formula (I) above wherein Rl and R2 each represents a hydrogen atomi R3 represents an acyl group having 2 to 40 carbon atoms; and 0~
R4 represents an alkyl group having 1 to 20 carbon atoms.
Particularly preferred compounds of the present invention are 2-acetamido-2-deoxy-1-S-decanoyl-3-O-(D-2-propanoyl-L-alanyl-D-isoglutamine methyl ester)-l-thio-~-D-glucopyranose, 2-acetamido-1-S-acetyl-2-deoxy-3-O-(D-
FIELD OF THE INVENTION
This invention relates to novel muramyldipeptide derivatives having excellent immunoadjuvant activity and prophylactic and therapeutic effects against microbial infections and antitumor activity and, more speclfically, this invention relates to a muramyldipeptide derivative represented by the formula ~
~ O~SR3 (I) R2~
NH--A c~ 1 CONH
C~3C~ICO-Ala-NH-C~CH2CH~COOR4 . I
wherein Ala represents alanine; Acyl represents an acyl group having 2 to 40 carbon atoms; ~1 and R2 each represents a hydrogen atom, or Rl and R2, when taken together, may : form an alkylidene group having 1 to 6 carbon atoms; R3 represents a hy~rogen atom, an acyl group having 2 to 40 carbon atoms or an alkyl group having 1 to 40 carbon atoms;
and R4 represents a hydrogen atom or an alkyl gxoup having : 1 to 20 carbon atoms.
^' ~
3~
BACKGROUND OF THE INVENTION
Hitherto, certain types of muramyldipeptide derivatives have been known to have useful immunotherapeutic activities. For example, U.S. Paten-t 4,101,536 discloses muramyldipeptide derivatives and the salts thereof having potent immunoadjuvant activi-ty and antltumor activity and being applicable as an agent for the immunotherapy of cancer for human and animals~ These known compaunds, however, have a free hydroxy group at the l-position of muramic acid of M-acetylmuramyldipeptide and do not possess an S-substituted moiety at the.l-position.
. SIJMMARY O`F THE INVENTION
As a result of extensive studies on compounds useful as adjuvant substances, we have found that the compounds of the above formula (I) which are obtained by introducing a sul~ur atom to ~he l-position of muramic acid o N-acylmuramyldipeptides, the minimum unit consti-tuting bact~rial cell wall of human type Mycobacterium tubercul_sis, BCG and other mycobacteria as well as cellular parastic bacteria, have excellent immunoadjuvant activity, and thus completed the present invention.
The compounds according to the present invention are of low toxicity and possess excellent immunoadjuvant activity and, therefore, can be useful for various diseases attributed to reduction in i~munity function~ for example, ., , O~
as prophylactic and therapeutic ayents against microbial infectlons, antitumor agents and the like.
DETAILED DESCRIPTION OF THE INVENTION
The muramyldipeptide derivatives of the present s invention are represented by the formula (I) ~ 9R3 R20 \~ ~
~ ~H-Acyl fONH2 CH3CHCO-Ala-NEI-CHCH2CH2COOR~
wherein Ala represents alanine; Acyl represents an acyl group having 2 to 40 carbon atoms; Rl and R2 each represents a hydrogen atom, or Rl and R2, when taken together, may : 10 form an alkylidene group having 1 to 6 carbon atoms; R3 represents a hydrogen atom, an acyl group having 2 to 40 carbon atoms or an alkyl group having 1 to 40 carbon atoms;
and R4 represents a hydrogen atom or an alkyl group having 1 to 20 carbon atoms.
lS A preferred class of compounds according to the present invention is that having the formula (I) above wherein Rl and R2 each represents a hydrogen atomi R3 represents an acyl group having 2 to 40 carbon atoms; and 0~
R4 represents an alkyl group having 1 to 20 carbon atoms.
Particularly preferred compounds of the present invention are 2-acetamido-2-deoxy-1-S-decanoyl-3-O-(D-2-propanoyl-L-alanyl-D-isoglutamine methyl ester)-l-thio-~-D-glucopyranose, 2-acetamido-1-S-acetyl-2-deoxy-3-O-(D-
2-propanyl-L-alanyl-D-isoglutamine methyl ester)-l-thio-~-D-glucopyranose, 2-acetamido-2-deoxy-3-O-(D-2-propanoyl~
L-alanyl-D-isoglutamine methyl ester)-l-thio-~-D-glucopyranose, 2-acetamido-2-deoxy-3-O-(D-2-propanoyl-L-alanyl-D-iso-glutamine)-l-thio-~-D-glucopyranose, 2-acetamido-2-deoxy-l-S-hexadecanoyl-3-O-(D-2-prop~noyl-L-alanyl-D-isoglutamine methyl ester)-l-thio-~-D-glucopyranose, 2-acetamido-2-deoxy-l-S-hexadecanoyl-3-O-(D-2-propanoyl-1-alanyl-D-iso-; glutamine)-l-thio-~-D-glucopyranose, and 2-acetamido-2-deoxy-1-S-eicosanoyl-3 O-(D-2-propanoyl-L-alanyl-D-iso-glutamine)-l-thio-~-D-glucopyranose.
The effects of the compounds of -the present invention were confirmed by the following method.
(1) Adjuvant Activity The adjuvant activity was confirmed by the enhancing activity on the induction of delayed-type hypersensitivity to N-acetyltyrosine-3-azobenzene-4'-arsonic acid (herein-after referred to as ABA-N-Ac-Tyr) in guinea pig.
That is, a water-in-oil emulsion prepared by 25 mixing 50 ~g/animal of ABA-N-Ac-Tyr and 100 ~g/animal of ~ S)~3 each of the compounds of the present invention with Freund's incomplete adjuvant ~hereinafter referred to as FIA) was administered to the soles of guinea pigs.
Two weeks later, 100 ~g of 3-azobenzene-4'-arsonic acid bonded to bovine serum albumin (hereinafter referred to as ABA-BSA3 was intradermally injected and the diameter of skin reaction (i.e., erythema and induration) was measured 24 hours and 48 hours after the injection. The diameter of the skin reaction is considered a measure of cellular immunity. The results obtained are shown in Table 1 below.
Table Adjuvant Activity on Induction of - Delayed-Type Hypersensitivity Skin Reaction Compound No.* Dose 24 Hours 48 Hours _ _ (~g~ (mm+SE) (mm+SE) 2 10021.0+1.0 21.0+0.7
L-alanyl-D-isoglutamine methyl ester)-l-thio-~-D-glucopyranose, 2-acetamido-2-deoxy-3-O-(D-2-propanoyl-L-alanyl-D-iso-glutamine)-l-thio-~-D-glucopyranose, 2-acetamido-2-deoxy-l-S-hexadecanoyl-3-O-(D-2-prop~noyl-L-alanyl-D-isoglutamine methyl ester)-l-thio-~-D-glucopyranose, 2-acetamido-2-deoxy-l-S-hexadecanoyl-3-O-(D-2-propanoyl-1-alanyl-D-iso-; glutamine)-l-thio-~-D-glucopyranose, and 2-acetamido-2-deoxy-1-S-eicosanoyl-3 O-(D-2-propanoyl-L-alanyl-D-iso-glutamine)-l-thio-~-D-glucopyranose.
The effects of the compounds of -the present invention were confirmed by the following method.
(1) Adjuvant Activity The adjuvant activity was confirmed by the enhancing activity on the induction of delayed-type hypersensitivity to N-acetyltyrosine-3-azobenzene-4'-arsonic acid (herein-after referred to as ABA-N-Ac-Tyr) in guinea pig.
That is, a water-in-oil emulsion prepared by 25 mixing 50 ~g/animal of ABA-N-Ac-Tyr and 100 ~g/animal of ~ S)~3 each of the compounds of the present invention with Freund's incomplete adjuvant ~hereinafter referred to as FIA) was administered to the soles of guinea pigs.
Two weeks later, 100 ~g of 3-azobenzene-4'-arsonic acid bonded to bovine serum albumin (hereinafter referred to as ABA-BSA3 was intradermally injected and the diameter of skin reaction (i.e., erythema and induration) was measured 24 hours and 48 hours after the injection. The diameter of the skin reaction is considered a measure of cellular immunity. The results obtained are shown in Table 1 below.
Table Adjuvant Activity on Induction of - Delayed-Type Hypersensitivity Skin Reaction Compound No.* Dose 24 Hours 48 Hours _ _ (~g~ (mm+SE) (mm+SE) 2 10021.0+1.0 21.0+0.7
3 10022.1+0.9 20.9+0.7
4 10023.1+1.2 21.0-~1.3 8 10014.5 ~a ~ 710.3+1.0 9 10013.5+0.8 12.4+0.7 10~17.4+1.7 13.6+1.8 11 10015.6+1.9 11.6+1.4 14 10020.4+0.7 21.0+1.4 - 25 15 10016.9+0.8 14.0+1.0 16 10011. 3+n ~ 85.6+1.4 17 100~1.8+0.8 23.1+1.1 Control ~ABA-Tyr+FIA) 4.5~0.3 2.9+0.7 o~
(2) Prophylactic ~ffect Against Microbial Infection Animals: Groups of 20 STD-ddy male mice, 5 weeks of age, were used.
Bac-teria: E. coli E77156 which has been maintained in a freezed-culture was used.
A~ents: Compounds of the present invention were dissolved or suspended in Dulbecco phosphate buffered saline (PBS: pH 7.4) at a concen-tration of 500 ~g/ml just before use.
Treatment and Infection:
The animals weEe treated subcutaneously with 0.2 ml of each of the above solution or suspension (equivalent to 100 ~g/mouse), follow~d by the subcutaneous challenge with E. coli 24 hours later.
The effect was judged from the percent survival o~ the mice seven days after the infection. As is apparent Erom the results shown in Table 2 belowl the compounds of the present invention exhibit excellent prophylactic effect '20 against microbial infection.
. . ..
, ~ ~
c)~
Table 2 Prophylactic Effect in Mice Infected with E. coli E 77156 ~ Survival 7 days after Infection Inoculum Size (cells/mouse) of E. coli E 77155 . _ Compound Number of 5No. Mice Used 4.~ x lo66.17 x 106 6 88 x lo6 : 24 20 65 PBS con~rol 20 5 5 5 (3) Antitumor Activity Against Meth-A System A mixture of tumor cells (~eth-A, 1.5 x 10 6 ) and 100 ~g of Compound 20 of -the present invention suspended in PBS was inoculated intradermally in BALB/c mice.
Suppress.ion of Meth-A growth in BALB/c on the 27th day after ;
:`
,~ , o~
inoculation was found to be 5/9. The value is expressed in te.rms of the number of tumor-free mice/the number of mice tested.
The test compounds used i'n the above experiments (1), (2) and (3) are as follows.
Compound 2: 2-Acetamido-l-S-acetyl-2-deoxy-3-O-(D-2-propanoyl-L-alanyl-D-isoglutamine methyl ester)-l-thio-~-D-glucopyranose Compound 3: 2-Acetamido-2-deoxy-3-O-(D-2-propanoyl-L-alanyl-D-isoglutamine methyl ester)-l-thio-~-D-glucopyranos~
Compound 4- 2-~cetamido-2-deoxy-3-O-(D-2-propanoyl-L-aranyl-D-isoglutamine)-l-thio-~-D-glucopyranose Compound 8: l-S-n-Butyl 2-acetamido-2-deoxy-3-O-(D-2-propanoyl-L-alanyl-D-isoglutamine methyl ester)-l-thio-~-D-glucopyranoside Compound 9: l-S-Hexadecanyl 2-acetamido-2-deoxy-3-O-(D-2-propanoyl-L alanyl-D-isoglutamine methyl ester)-l-thio-~-D-glucopyranoside Compound 10: l-S-n-Butyl 2-acetamido-2-deoxy-3-O-(D-2-. propanoyl-L-alanyl-D~isoglutamine)-l-thio-~-D-glucopyranoside Compound 11: l-S-Hexadecanyl 2-acetamido-2-deoxy-3-O-(D-2-propanoyL-L-alanyl-D-isoglutamine)-l-thi ~-D-glucopyranoside - ~ _ :, :
i o~
Compound 14: 2-Acetamido~2-deoxy-1-S-hexadecanoyl-3-O-(D-2-propanoyl-L-alanyl-D-isoglutamine methyl ester)-l-thio-~-D-glucopyranose Compound 15: 2-Acetamido-2-deoxy-4,6-O-isopropylidene-3-O-(D-2-propanoyl-L-alanyl-D-isoglutamine)-l-thio-~-D-glucopyranose Compound 16: 2-Acetamido-2-deoxy-1-S-hexadecanoyl-4,6-O-isopropylidene-3-O-(D-2-propanoyl-L-alanyl-D-isoglutamine)-l-thio-~-D-glucopyranose Compound 17: 2-Acetamido-2-deoxy-1-S-hexadecanoyl-3-O-(D-2-propanoyl-L.alanyl-D-isoglutamine)-l-thio-~-D-glucopyranose Compound 18: 2-Acetamido-2-deoxy-1-S-decanoyl-3-O-(D-2-propanoyl-L-alanyl-D-isoglutamine methy].
ester)-l-thio-~-D-glucopyrano5e Compound 19: 2-Acetamido-2-deoxy-1-S-octadecanoyl-3-O-(D-2-propanoyl-L-alanyl-D-isoglutamine me~hyl ester)-l-thio-~-D-glucopyranose Compound 20: 2-Acetamido-2-deoxy-1-S-triacontanoyl-3-O-(D-2-propanoyl-L-alanyl-D isoglutamine methyl ester)-l-thio-~-D-glucopyranose Compound 21: 2-Acetamido-2-deoxy-1-S-n-butanoyl-3-O-(D-2-propanoyl-L-alanyl-D-isoglutamine methyl ester)-l-thio-~-D-glucopyranose .
~L2~40C~
Compound 220 2 Acetamido-2-deoxy-1-S-dodecanoyl-3-0-(D-2-propanoyl-L-alanyl-D-isoglutamine)-l-thio-~-D-glucopyranose.
Compound 23: 2-Acetamido-2-deoxy-1-S-tetradecanoyl-3-0-(D-2-propanoyl-L-alanyl-D-isoglutamine) 1-thio-~-D-glucopyranose Compound 24: 2-Acetamido-2-deoxy-1-S-tetradecanoyl-3-0-(D-2-propanoyl-L-alanyl-D-isoglutamine methyl ester)-l-thio-~-D-glucopyranose.
Compound 25: 2-Acetamido-2-deoxy-1-S-eicosanoyl-3-0-(D-2-propanoyl-L-alanyl-D-isoglutamine)-l-thio-~-D-glucopyranose.
The compounds of the present invention can be prepared according to the following reaction scheme.
- .
R5 X ~ C~2- rO ) R6 ~ ~
NHAcyl NHAcyl CH3cHcooR7 ~ CH3cHcooR7 (II) (III) 12~
R5~<OC'~ ~ RSX~CH ~
NHAcyl NHAcyl ~IV) (V) R~<o/~o ~ ~ R6>o~¢
NHAcy 1 I NHAcyl CH3cHcooR7 (VI) (VII) > ( I ) I
I' ;3~0~
In the above reaction scheme, R5 represents a hydrogen atom or an alkyl group having 1 to 6 carbon atom, and R6 and R7 each represents an alkyl group having 1 to 6 carbon atom.
That is, the compound (II) is oxidized in the presence of a solvent, such as dichloromethane, using a chromic anhydride-pyridine complex at a temperature of about 0C to 60C to obtain the compound (III).
The compound (III) is reacted with a metal alcoholate in an alcohol solvent to obtain the compound (IV). The compound (IV) is dissolved in a solvent, such as a mixed . solvent of dichloromethane and carbon tetrachloride, and then reacted with hexamethylphosphorus triamide [P(NMe2)3]
at a low temperature, preferably -40C or lower, to obtain the compound (V). The compound (V) is dissolved in a solvent or a mixture of solvents, for example, a mixture of dichloromethane and acetone, and reacted wi-th an alkali thio-organic acid, for example, potassium thioacetate (C~3COSK), to obtain the compound (VI)~ The compound (VI) is reacted with an alkali hydroxide in the presence of a solvent, such as dioxane, to hydrolyze the S-acetyl group and alkoxycarbonyl group and then the hydrolysate is reacted with an organic acid anhydride such as acetic anhydride, and triethylamine to acylate the SH group thereby obt.aining the compound (VII).
'~
L~Lo~i The compound (VII) -thus obtained is then condensed with an alanyl-isoglutamine alk~l ester to obtain the desired compound of the formula (I) wherein Rl and R2 are bonded together to form an alkylidène group; R3 is an acyl group; and R4 is an alkyl group.
The above condensation reaction can be carried out by methods commonly employed for peptide syntheses, i.e., carbodiimide method, eintops method, active ester method, acid anhydride method and the like. For example, the compound (VII) is dissolved in a solvent such as dioxane, and dicyclohexylcarbodiimide and N-hydroxysuccinic acid imide or l-hydroxybenzotriazole are added to the solution at about 0C to 60C to form an active ester of the compound (~II). To the resulting reaction mixture is added an L-alanyl-D-isoglutamine alkyl ester at about 0C
to 60C to obtain the above-described compound (I).
The S-acyl group of the compound (I) thus obtained can be converted into a free SH group by reacting the compound (I) with a metal alcoholate in an alcoholic solution at about 0C to room tempera~ure, followed by neutralization with a cation exchange resin, etc.
The thus obtained SH-compound may be converted into an S-acylated compound by reacting with an acyl halide in a solvent, such as dichloromethane, in the presence of an acid acceptor such as pyridine, at about -10C to 50C.
$ L~ O~
Further, the S~alkylated compound can be obtained by reacting the above S-acylated compound with a sodium alcoholate in an alcohol and, without conducting neutralization, the resulting product is then xeacted with an alkyl halide at 0 to 40C. The alkylidene group can be released by reacting with a 50 - 90% aqueous solution of acetic acid at room temperature to 100C. The conversion of the alkoxycarbonyl group to a carboxyl group can be accomplished by reacting with an aqueous alkali hydroxide solution in an alcoholic solution at 0 to room temperature and then neutralizing with a cation exchange resin, etc.
The present in~ention will now be described in greater detail with reference to examples but the scope of the invention is not limited thereto. Unless otherwise indicated, a mixture of solvents is by volume.
Example (1) 164.7 mg of chromic anhydride and 0.25 ml of pyridine were added to 5 ml of dichloromethane and the mixture was stirred for 15 minutes. To the resulting mixture were added 180 mg of benzyl 2-acetamido-2-deoxy~
4,6-O-isopropylidine-3-~-[D-l-(methoxycarbonyl)ethyl] a-D-glucopyranoside (compound II) dissolved in 3 ml of dichloromethane and then 0.16 ml of acetic anhydride.
-The reaction mixture was stirred at 45C for 3 hours and passed through a column of silica gel (Wako~C-r~
:"
~,z~
200, 20 g), and the column was eluted with ethyl acetate to obtain 170 mg (yield: 92%) of 2-acetamido-1-o-benzoyl-2-deoxy-4,6 O-isopropylidene 3-o-[D-l-(methoxycarbonyl)-ethyl]-~-D-glucopyranose (compound III) having a melting point of 134-137C.
[~]25 +144 (c=1.5, methanol) Elemental Analysis for C22H29NOg:
Calcd. (%): C, 58.53; H, 6.47; N, 3.10 Found (%): C, 58.39; H, 6.58; N, 3.02 (2) To a solution of 4.0 g of the compound (III) in 60 ml of absolute methanol, about 30 mg of metai sodium was added under cooling. After allowing to stand at room temperature for 10 minutes, the mixture was treated with Amberlite IRC-50. The solution was concentrated and the resulting syrup was purified by column chromatography using silica gel (50 g) (developing agent (a): chloroform, developiny agent (b): chloroform-methanol = 50:1).
From the eEfluent (b), there was obtained 2.8 g ~yield: 93%) of 2-acetamido-2-deoxy-4,6-O-isopropylidene-3-O-~D-l-(methoxy-carbonyl)ethyl]-D-glucopyranose (compound IV) having a melting point of 180 - 184C.
[a]D5 -t44.3O (c=0.47, chloroform, equilibrium) Elemental Analysis for C15H25NO8:
Calcd. (%): C, 51.86; H, 7.25; N, 4.03 ; 25 Found (%): C, 51.81; H, 7.29; N, 3.96 -rf2 ,9 D ~ ~R ~ -9~
(3) 2.5 g of the compound (IV) was dissolved in a mixture of 50 ml of anhydrous dichloromethane and 2.17 g of anhydrous carbon tetrachloride and the mixture was cooled to -50C~ To the resulting mixture was added a solution of 1.6 g of hexamethylphosphorus triamide in 25 ml of anhydrous dichloromethane over 15 minukes while stirring at -50C, and the reaction was conducted for 1 hour. The reaction mixture was concentrated under reduced pressure (bath temperature: 40C). At this stage, the thin layer chromatography revealed the presence of a mix-ture of ~- and ~-chlorides and oxazoline. By repeating the above procedure of addition of anhydrous dichloromethane followed by concentration three to four times, all the chlorides were led to the a-chloro derivative. The a-chloride was dissolved in a mixed solution of 15 ml ofanhydrous dichloromethane and 15 ml of anhydrous acetone, a~d 2.4 g of potassium thioacetate was added thereto, followed by stirring the mixture at room temperatLlre overnight. After it was confirmed that all the chloride disappeared by thin la~er chromatography, the reaction ; mLxture was conc~ntrated under reduced pressure, and the residue was extracted with chloroform, washed with water, and dried over sodium sulfate. The solvent was removed, and the residue was crystallized from diethyl ether to 25 obtain 1.94 g (yield: 66.4%~ of 2-acetamido-1-S-acetyl-- 16 ~
:,., "
~2~
2-deoxy-4,6-O-isopropylidene-3-O-[D-l-(methoxycarbonyl)-ethyl]-l-thio-~-D-glucopyranose (compound VIj as a needle-like crystal. The filtxate was purified by silica gel (30 g) column chromatography (developing agent (a): chloroform;
(b): chloroform-methanol = 150:1). From the effluent (b) was obtained 500 mg (yield: 17.1%) of the compound ~VI).
Total yield: 2.44 g (83.5~); melting point: 171 - 173C.
[~]D +8.1 (c=0.4, chloroform) Elemental Analys~s for C17H27NO8S:
Calcd. (~): C, 50.36; H, 6.71; N, 3.45 Found (%): C, 50.28.; H, 6.59; N, 3.45 (4) 200 mg.of the compound (VI) was dissolved in 10 ml of 1,4-dioxane, and 12 ml of 0.2M potassium hydroxide was added thereto. After allowing the mixture to stand at room temperakure for 10 minutes, the mixture was treated with Amberlite IRC-50 to remove the base. The resin was separated by filtration and washed with methanol.
The filtrate and the washing were combined and concentrated under reduced pressure. The resulting syrup was dissolved 2~ in methanol, and 1 ml of triethylamine and 0.8 ml of anhydrous acetic acid were added to the solution under cooling, followed by reacting at room temperature for 30 minutes. The reaction mixture was concentrated under reduced pressure, and the resulting syrup was purified by silica gel (20 g) column chromatography ~developing .
..
~.2'~3 ?*~)~
agent (a): chloroform; (b): chloroform-methanol=50:1).
From the effluent (b), there was obtained 150 mg (yield: 78%) of 2-acetamido-1-S acetyl-3-O-(D-l-carboxyethyl)-2 deoxy-4,6-O-isopropylidene-l-tllio-a-D-glucopyranose (compound VII) as needle-like crystals. Recrystallization from diethyl ether gave a product having a melti,ng point of 193 - 200C (decomposition).
~a]D +10.5 (c=0.3, chloroform~
Elemental Analysis for C16H25NO8S:
Calcd. (~): C, 49.09; H, 6.44; N, 3.58 Found (%): C, 49.13; H, 6.51; N, 3.48
(2) Prophylactic ~ffect Against Microbial Infection Animals: Groups of 20 STD-ddy male mice, 5 weeks of age, were used.
Bac-teria: E. coli E77156 which has been maintained in a freezed-culture was used.
A~ents: Compounds of the present invention were dissolved or suspended in Dulbecco phosphate buffered saline (PBS: pH 7.4) at a concen-tration of 500 ~g/ml just before use.
Treatment and Infection:
The animals weEe treated subcutaneously with 0.2 ml of each of the above solution or suspension (equivalent to 100 ~g/mouse), follow~d by the subcutaneous challenge with E. coli 24 hours later.
The effect was judged from the percent survival o~ the mice seven days after the infection. As is apparent Erom the results shown in Table 2 belowl the compounds of the present invention exhibit excellent prophylactic effect '20 against microbial infection.
. . ..
, ~ ~
c)~
Table 2 Prophylactic Effect in Mice Infected with E. coli E 77156 ~ Survival 7 days after Infection Inoculum Size (cells/mouse) of E. coli E 77155 . _ Compound Number of 5No. Mice Used 4.~ x lo66.17 x 106 6 88 x lo6 : 24 20 65 PBS con~rol 20 5 5 5 (3) Antitumor Activity Against Meth-A System A mixture of tumor cells (~eth-A, 1.5 x 10 6 ) and 100 ~g of Compound 20 of -the present invention suspended in PBS was inoculated intradermally in BALB/c mice.
Suppress.ion of Meth-A growth in BALB/c on the 27th day after ;
:`
,~ , o~
inoculation was found to be 5/9. The value is expressed in te.rms of the number of tumor-free mice/the number of mice tested.
The test compounds used i'n the above experiments (1), (2) and (3) are as follows.
Compound 2: 2-Acetamido-l-S-acetyl-2-deoxy-3-O-(D-2-propanoyl-L-alanyl-D-isoglutamine methyl ester)-l-thio-~-D-glucopyranose Compound 3: 2-Acetamido-2-deoxy-3-O-(D-2-propanoyl-L-alanyl-D-isoglutamine methyl ester)-l-thio-~-D-glucopyranos~
Compound 4- 2-~cetamido-2-deoxy-3-O-(D-2-propanoyl-L-aranyl-D-isoglutamine)-l-thio-~-D-glucopyranose Compound 8: l-S-n-Butyl 2-acetamido-2-deoxy-3-O-(D-2-propanoyl-L-alanyl-D-isoglutamine methyl ester)-l-thio-~-D-glucopyranoside Compound 9: l-S-Hexadecanyl 2-acetamido-2-deoxy-3-O-(D-2-propanoyl-L alanyl-D-isoglutamine methyl ester)-l-thio-~-D-glucopyranoside Compound 10: l-S-n-Butyl 2-acetamido-2-deoxy-3-O-(D-2-. propanoyl-L-alanyl-D~isoglutamine)-l-thio-~-D-glucopyranoside Compound 11: l-S-Hexadecanyl 2-acetamido-2-deoxy-3-O-(D-2-propanoyL-L-alanyl-D-isoglutamine)-l-thi ~-D-glucopyranoside - ~ _ :, :
i o~
Compound 14: 2-Acetamido~2-deoxy-1-S-hexadecanoyl-3-O-(D-2-propanoyl-L-alanyl-D-isoglutamine methyl ester)-l-thio-~-D-glucopyranose Compound 15: 2-Acetamido-2-deoxy-4,6-O-isopropylidene-3-O-(D-2-propanoyl-L-alanyl-D-isoglutamine)-l-thio-~-D-glucopyranose Compound 16: 2-Acetamido-2-deoxy-1-S-hexadecanoyl-4,6-O-isopropylidene-3-O-(D-2-propanoyl-L-alanyl-D-isoglutamine)-l-thio-~-D-glucopyranose Compound 17: 2-Acetamido-2-deoxy-1-S-hexadecanoyl-3-O-(D-2-propanoyl-L.alanyl-D-isoglutamine)-l-thio-~-D-glucopyranose Compound 18: 2-Acetamido-2-deoxy-1-S-decanoyl-3-O-(D-2-propanoyl-L-alanyl-D-isoglutamine methy].
ester)-l-thio-~-D-glucopyrano5e Compound 19: 2-Acetamido-2-deoxy-1-S-octadecanoyl-3-O-(D-2-propanoyl-L-alanyl-D-isoglutamine me~hyl ester)-l-thio-~-D-glucopyranose Compound 20: 2-Acetamido-2-deoxy-1-S-triacontanoyl-3-O-(D-2-propanoyl-L-alanyl-D isoglutamine methyl ester)-l-thio-~-D-glucopyranose Compound 21: 2-Acetamido-2-deoxy-1-S-n-butanoyl-3-O-(D-2-propanoyl-L-alanyl-D-isoglutamine methyl ester)-l-thio-~-D-glucopyranose .
~L2~40C~
Compound 220 2 Acetamido-2-deoxy-1-S-dodecanoyl-3-0-(D-2-propanoyl-L-alanyl-D-isoglutamine)-l-thio-~-D-glucopyranose.
Compound 23: 2-Acetamido-2-deoxy-1-S-tetradecanoyl-3-0-(D-2-propanoyl-L-alanyl-D-isoglutamine) 1-thio-~-D-glucopyranose Compound 24: 2-Acetamido-2-deoxy-1-S-tetradecanoyl-3-0-(D-2-propanoyl-L-alanyl-D-isoglutamine methyl ester)-l-thio-~-D-glucopyranose.
Compound 25: 2-Acetamido-2-deoxy-1-S-eicosanoyl-3-0-(D-2-propanoyl-L-alanyl-D-isoglutamine)-l-thio-~-D-glucopyranose.
The compounds of the present invention can be prepared according to the following reaction scheme.
- .
R5 X ~ C~2- rO ) R6 ~ ~
NHAcyl NHAcyl CH3cHcooR7 ~ CH3cHcooR7 (II) (III) 12~
R5~<OC'~ ~ RSX~CH ~
NHAcyl NHAcyl ~IV) (V) R~<o/~o ~ ~ R6>o~¢
NHAcy 1 I NHAcyl CH3cHcooR7 (VI) (VII) > ( I ) I
I' ;3~0~
In the above reaction scheme, R5 represents a hydrogen atom or an alkyl group having 1 to 6 carbon atom, and R6 and R7 each represents an alkyl group having 1 to 6 carbon atom.
That is, the compound (II) is oxidized in the presence of a solvent, such as dichloromethane, using a chromic anhydride-pyridine complex at a temperature of about 0C to 60C to obtain the compound (III).
The compound (III) is reacted with a metal alcoholate in an alcohol solvent to obtain the compound (IV). The compound (IV) is dissolved in a solvent, such as a mixed . solvent of dichloromethane and carbon tetrachloride, and then reacted with hexamethylphosphorus triamide [P(NMe2)3]
at a low temperature, preferably -40C or lower, to obtain the compound (V). The compound (V) is dissolved in a solvent or a mixture of solvents, for example, a mixture of dichloromethane and acetone, and reacted wi-th an alkali thio-organic acid, for example, potassium thioacetate (C~3COSK), to obtain the compound (VI)~ The compound (VI) is reacted with an alkali hydroxide in the presence of a solvent, such as dioxane, to hydrolyze the S-acetyl group and alkoxycarbonyl group and then the hydrolysate is reacted with an organic acid anhydride such as acetic anhydride, and triethylamine to acylate the SH group thereby obt.aining the compound (VII).
'~
L~Lo~i The compound (VII) -thus obtained is then condensed with an alanyl-isoglutamine alk~l ester to obtain the desired compound of the formula (I) wherein Rl and R2 are bonded together to form an alkylidène group; R3 is an acyl group; and R4 is an alkyl group.
The above condensation reaction can be carried out by methods commonly employed for peptide syntheses, i.e., carbodiimide method, eintops method, active ester method, acid anhydride method and the like. For example, the compound (VII) is dissolved in a solvent such as dioxane, and dicyclohexylcarbodiimide and N-hydroxysuccinic acid imide or l-hydroxybenzotriazole are added to the solution at about 0C to 60C to form an active ester of the compound (~II). To the resulting reaction mixture is added an L-alanyl-D-isoglutamine alkyl ester at about 0C
to 60C to obtain the above-described compound (I).
The S-acyl group of the compound (I) thus obtained can be converted into a free SH group by reacting the compound (I) with a metal alcoholate in an alcoholic solution at about 0C to room tempera~ure, followed by neutralization with a cation exchange resin, etc.
The thus obtained SH-compound may be converted into an S-acylated compound by reacting with an acyl halide in a solvent, such as dichloromethane, in the presence of an acid acceptor such as pyridine, at about -10C to 50C.
$ L~ O~
Further, the S~alkylated compound can be obtained by reacting the above S-acylated compound with a sodium alcoholate in an alcohol and, without conducting neutralization, the resulting product is then xeacted with an alkyl halide at 0 to 40C. The alkylidene group can be released by reacting with a 50 - 90% aqueous solution of acetic acid at room temperature to 100C. The conversion of the alkoxycarbonyl group to a carboxyl group can be accomplished by reacting with an aqueous alkali hydroxide solution in an alcoholic solution at 0 to room temperature and then neutralizing with a cation exchange resin, etc.
The present in~ention will now be described in greater detail with reference to examples but the scope of the invention is not limited thereto. Unless otherwise indicated, a mixture of solvents is by volume.
Example (1) 164.7 mg of chromic anhydride and 0.25 ml of pyridine were added to 5 ml of dichloromethane and the mixture was stirred for 15 minutes. To the resulting mixture were added 180 mg of benzyl 2-acetamido-2-deoxy~
4,6-O-isopropylidine-3-~-[D-l-(methoxycarbonyl)ethyl] a-D-glucopyranoside (compound II) dissolved in 3 ml of dichloromethane and then 0.16 ml of acetic anhydride.
-The reaction mixture was stirred at 45C for 3 hours and passed through a column of silica gel (Wako~C-r~
:"
~,z~
200, 20 g), and the column was eluted with ethyl acetate to obtain 170 mg (yield: 92%) of 2-acetamido-1-o-benzoyl-2-deoxy-4,6 O-isopropylidene 3-o-[D-l-(methoxycarbonyl)-ethyl]-~-D-glucopyranose (compound III) having a melting point of 134-137C.
[~]25 +144 (c=1.5, methanol) Elemental Analysis for C22H29NOg:
Calcd. (%): C, 58.53; H, 6.47; N, 3.10 Found (%): C, 58.39; H, 6.58; N, 3.02 (2) To a solution of 4.0 g of the compound (III) in 60 ml of absolute methanol, about 30 mg of metai sodium was added under cooling. After allowing to stand at room temperature for 10 minutes, the mixture was treated with Amberlite IRC-50. The solution was concentrated and the resulting syrup was purified by column chromatography using silica gel (50 g) (developing agent (a): chloroform, developiny agent (b): chloroform-methanol = 50:1).
From the eEfluent (b), there was obtained 2.8 g ~yield: 93%) of 2-acetamido-2-deoxy-4,6-O-isopropylidene-3-O-~D-l-(methoxy-carbonyl)ethyl]-D-glucopyranose (compound IV) having a melting point of 180 - 184C.
[a]D5 -t44.3O (c=0.47, chloroform, equilibrium) Elemental Analysis for C15H25NO8:
Calcd. (%): C, 51.86; H, 7.25; N, 4.03 ; 25 Found (%): C, 51.81; H, 7.29; N, 3.96 -rf2 ,9 D ~ ~R ~ -9~
(3) 2.5 g of the compound (IV) was dissolved in a mixture of 50 ml of anhydrous dichloromethane and 2.17 g of anhydrous carbon tetrachloride and the mixture was cooled to -50C~ To the resulting mixture was added a solution of 1.6 g of hexamethylphosphorus triamide in 25 ml of anhydrous dichloromethane over 15 minukes while stirring at -50C, and the reaction was conducted for 1 hour. The reaction mixture was concentrated under reduced pressure (bath temperature: 40C). At this stage, the thin layer chromatography revealed the presence of a mix-ture of ~- and ~-chlorides and oxazoline. By repeating the above procedure of addition of anhydrous dichloromethane followed by concentration three to four times, all the chlorides were led to the a-chloro derivative. The a-chloride was dissolved in a mixed solution of 15 ml ofanhydrous dichloromethane and 15 ml of anhydrous acetone, a~d 2.4 g of potassium thioacetate was added thereto, followed by stirring the mixture at room temperatLlre overnight. After it was confirmed that all the chloride disappeared by thin la~er chromatography, the reaction ; mLxture was conc~ntrated under reduced pressure, and the residue was extracted with chloroform, washed with water, and dried over sodium sulfate. The solvent was removed, and the residue was crystallized from diethyl ether to 25 obtain 1.94 g (yield: 66.4%~ of 2-acetamido-1-S-acetyl-- 16 ~
:,., "
~2~
2-deoxy-4,6-O-isopropylidene-3-O-[D-l-(methoxycarbonyl)-ethyl]-l-thio-~-D-glucopyranose (compound VIj as a needle-like crystal. The filtxate was purified by silica gel (30 g) column chromatography (developing agent (a): chloroform;
(b): chloroform-methanol = 150:1). From the effluent (b) was obtained 500 mg (yield: 17.1%) of the compound ~VI).
Total yield: 2.44 g (83.5~); melting point: 171 - 173C.
[~]D +8.1 (c=0.4, chloroform) Elemental Analys~s for C17H27NO8S:
Calcd. (~): C, 50.36; H, 6.71; N, 3.45 Found (%): C, 50.28.; H, 6.59; N, 3.45 (4) 200 mg.of the compound (VI) was dissolved in 10 ml of 1,4-dioxane, and 12 ml of 0.2M potassium hydroxide was added thereto. After allowing the mixture to stand at room temperakure for 10 minutes, the mixture was treated with Amberlite IRC-50 to remove the base. The resin was separated by filtration and washed with methanol.
The filtrate and the washing were combined and concentrated under reduced pressure. The resulting syrup was dissolved 2~ in methanol, and 1 ml of triethylamine and 0.8 ml of anhydrous acetic acid were added to the solution under cooling, followed by reacting at room temperature for 30 minutes. The reaction mixture was concentrated under reduced pressure, and the resulting syrup was purified by silica gel (20 g) column chromatography ~developing .
..
~.2'~3 ?*~)~
agent (a): chloroform; (b): chloroform-methanol=50:1).
From the effluent (b), there was obtained 150 mg (yield: 78%) of 2-acetamido-1-S acetyl-3-O-(D-l-carboxyethyl)-2 deoxy-4,6-O-isopropylidene-l-tllio-a-D-glucopyranose (compound VII) as needle-like crystals. Recrystallization from diethyl ether gave a product having a melti,ng point of 193 - 200C (decomposition).
~a]D +10.5 (c=0.3, chloroform~
Elemental Analysis for C16H25NO8S:
Calcd. (~): C, 49.09; H, 6.44; N, 3.58 Found (%): C, 49.13; H, 6.51; N, 3.48
(5) 160 m~ of the compound (VII) was dissolved in 5 ml of anhydrous 1,4-dioxane, and 210 mg of dicyclohexyl-carbodiimide and 100 mg of N-hydroxysuccinimide were added to the solution, and the mixture was stirred at room temperature for 30 minutes. The diphenylurea formed was separated by filtration and washed with 2 ml of dioxane.
The filtrate and the washing were combined, and 200 mg of L-alanyl-D-isoglutamine methyl ester trifluoroacetate and 5 drops of triethylamine were added thereto while stirring to effect a reaction at room temperature for 40 minutes. After completion of the reaction was confirmed by thin layer chromatography, the reaction mixture was concentrated under reduced pressure, and the resulting syrup was purified by silica gel (20 g) column chromatograpy (developing agent (a): chloroform; (b): chloroform-methanol=
100:1; (c): chloroform-metAanol=50:1). From the ef~luent (c), there was obtained 235 mg (yield: 95%) of th~ desired compounds, 2-acetamido-1-S-acetyl-2-deoxy-4,6-O-isopropylidene-3-O-(D-2-propanoyl-L-alanyl-D-isoglutamine methyl ester)-l-thio-~-D-glucopyranose (compound I). Recrystallization from diethyl ether gave a product having a melting point of 148 - 151C.
~a]D +11 (c=0.2, chlorofoxm) Elemental Analysis for C25H40N4OllS:
Calcd. (%): C, 49.6~; H, 6.67; N, 9.27 Found (%): C, 49.76; H, 6.65; N, 9.30
The filtrate and the washing were combined, and 200 mg of L-alanyl-D-isoglutamine methyl ester trifluoroacetate and 5 drops of triethylamine were added thereto while stirring to effect a reaction at room temperature for 40 minutes. After completion of the reaction was confirmed by thin layer chromatography, the reaction mixture was concentrated under reduced pressure, and the resulting syrup was purified by silica gel (20 g) column chromatograpy (developing agent (a): chloroform; (b): chloroform-methanol=
100:1; (c): chloroform-metAanol=50:1). From the ef~luent (c), there was obtained 235 mg (yield: 95%) of th~ desired compounds, 2-acetamido-1-S-acetyl-2-deoxy-4,6-O-isopropylidene-3-O-(D-2-propanoyl-L-alanyl-D-isoglutamine methyl ester)-l-thio-~-D-glucopyranose (compound I). Recrystallization from diethyl ether gave a product having a melting point of 148 - 151C.
~a]D +11 (c=0.2, chlorofoxm) Elemental Analysis for C25H40N4OllS:
Calcd. (%): C, 49.6~; H, 6.67; N, 9.27 Found (%): C, 49.76; H, 6.65; N, 9.30
(6) 100 mg of the~compound (I) was dissolved in 5 ml ; of a 80% aqueous acetic acid solu-tion and heated at 45C
for 45 minutes while stirring. The reaction mixture was concentrated under reduced pressure at below 40C. The acetic acid was completely removed as an azeotropic mixture with toluene to obtain 2-acetamido-1-S-acetyl~2-deoxy-3-O-(D-2-propanoyl-L-alanyl-D-isoglutamine methyl ester)-l-thio-~-D-glucopyranose (Compound 2) that showed a single spot on the thin layer chromatogram. ~ecrys~allization from diethyl ether gave a pure product having a melting point of 158 - 162C.
[o~]D5 +62 (c=0.2, chloroform-methanol=l:l) Elemental Analysis for C22H36N4OllS:
Calcd. (%): C, 46.80; H, 6.43; N, 9.92 Found (%): C, 46.51; H, 6.65; N, 9.85 Example 2 20 mg of Compound 2 as prepared in Example 1-(6) was dissolved in 2 ml of absolute methanol, and a small amount of metal sodium was added thereto. Three minutes later, disappearance of any unreacted substances was 10 conEirmed by thin layer chromatography, and the reaction mixture was treated with Amberiite IR-120 (H ~ ollowed by filtration. From the filtrate was obtained 17.6 mg (yield: 95%) of 2-acetamido-2-deoxy-3-O (D-2-propanoyl-L-alanyl-D-isoglutamine methyl ester)-l-thio-~-D-glucopyranose (Compound 3) having a melting point of 117 - 125C
(decomposition).
[a]D5 ~24 (c=0.2, methanol, equilibrium) Elemental Analysis for C20H34N4OloS
Calcd. (g6): C, 45.97; H, 6.56; N, 10.72 20 Found (~): C, 45.49, H, 6.53, N, 10.88 Example 3 25 mg of Compound 2 as prepared in Example 1-(6) was dissolved in 2 ml of absolute methanol, and a small amount of metal sodium was added thereto. Three minutes ,, ~ :
: :, :",:
later, 1 ml of 0.2M potassium hydroxide was added to the reaction mixture. After confirming completion of the reaction by thin layer chromatography, the reaction mixture was treated with Amberlite [R-120 (H ) to remove the base. The solution was concentrated under reduced pressure and freeze-dried from 1,4-dioxane to obtain 22.4 mg (quantitative yield) of 2-acetamido-2-deoxy-3-O-(D-2-propanoyl-L-alanyl-D-isoglutamine)-l-thio-~-D-glucopyranose (Compound 4) having a melting point of 157 -166C (decomposition).
[~]25 -~20 (c=0.2, methanol, e~uilibrium) Elemental Analysis for ClgH32N4OloS:
Calcd. (%): C, 44.87; H, 6.34; N, 11.02 Found (~): C, 44-.45; H, 6.53; N, 10.99 Example 4 60 mg of Compound 1 as prepared in Example 1-(5) was dissolved in 5 ml of absolute methanol, and 6 mg of sodium mekhoxide was added thereto. Ninety minutes later, a~ter it was confirmed by thin layer chromatoyraphy tha-t the reactant had completely be converted to 2-acetamido-l-S-sodium-2-deoxy-4,6-O-isopropylidene-3-O-(D-2-propanoyl-L-alanyl-D~isogulutamine methyl ester)-1-thio-~-D-glucopyranose (Compound 5), 15 mg of n-butyl bromide was added to the reaction mixture, followed by stirring at room temperature overnight. The reaction mixture was concentrated under :
~ .
o~
reduced pressure, and the resulting syrup was purified by silica gel (10 g) column chromatography (developing agent (a): chloroform; (b): chloroform-methanol=50:1) to obtain 60 mg (yield: 98%) of l-S-n-butyl 2 acetamido-2-deoxy-4,6-O-isopropylidene-3-O-(D-2-propanoyl-L-alanyl-D-isoglutamine methyl ester)-l-thio-~-D-glucopyranoside (Compound 6) having a melting point of 75 - 78C.
[a]25 +25 ~c=0.5, methanol) Elemental Analysis for C27H46N4OloS:
Calcd. (%): C, 52.41; H, 7.49; N, 9.06 Found (%): C, 52.25; H, 7.45; N, 9.01 ~ Exampl`e 5 Compound 5 was prepared in the same manner as in Example 4 but using 83 mg of Compound 1. 43 mg of hexadecanyl bromide was added to the reaction mixture containing Compound 5, and the resulting mixture was stirred at room temperature overnight. The reaction mixture was concentrated under reduced pressure and then purified by silica gel (lO g) column chromatography (developing agent (a): chloroform-methanol=100:1, (b): chloroform-methanol=30:1). From the effluent (b)~ there was obtained lO0 mg (yield: 93%) of l-S-hexadecanyl 2-acetamido-2-deoxy-4,6-O-isopropylidene-3-O-(D-2-propanoyl-L-alanyl-D-isoglutamine methyl ester)-l-thio-3-D-glucopyranoside (Compound 7) having a melting ~.2~
point of 93 - 96C.
[a]D +35 (c=0.3, chloroform) Elemental Analysis for C39H70N4OloS
Calcd. (%): C, 59.51; H; 8.97; N, 7.17 Found (%): C, 59.33; H, 8.86; N, 7.15 Example 6 50 mg of Compound 6 as prepared in Example 4 was dissolved in 3 ml of a 8% aqueous acetic acid solution, and hydrolysis was carried out at 45C for 60 minutes.
After confirming by thin layer chromatography, the reaction mixture was freeze-dried to quantitatively obtain l-S-n-butyl 2-acetamido-2-deoxy-3-O-(D-2-propanoyl-L-alanyl-D-isoglutamine methyl ester) l-thio-~-D-glucopyranoside (Compound 8) having a melting point of 155 - 160C
(decomposition).
[a]D ~14 (c=0.3, methanol) Elemental Analysis for C24H42N4OllS:
Calcd. (%): C, 49.81; H, 7.32; N, 9.68 Found (~): C, 49.45; H, 7.61; N, 9.55 Example 7 50 mg of Compound 7 as prepared in Example 5 was dissolved in 3 ml of a 80% aqueous acetic acid solution and heated at 45C for 1 houx. After confirming disappearance of any unreacted substances by thin layer - 23 ~
chromatography, the reaction mixture was freeze-dried to obtain l-S-hexadecanyl 2-acetamido-2-deoxy-3-O-(D-2-propanoyl-L-alanyl-D-isoglutamine methyl ester)-l-thio-~-D-glucopyranoside (Compound 9) having a melting point of 179 - 182C in a quantitative yield.
- [~]D5 +44 (c=0.2, chloroform-methanol=l:l) Elemental Analysis for C36H66N4OloS:
Calcd. (~): C, 57.88; H, 8.91; N, 7.51 Found (%): C, 57.65; H, 8.96; N, 7.43 Example 8 20 mg of Compound 8 as prepa~ed in Example 6 was dissolved in 2 ml of methanol, and 2 ml of 0.2M
potassium hydxoxide was added thereto. Twenty minutes later, the ester was completely hydrolyzed. The reaction ; 15 mixture was treated with Amberlite IR-120 (H ), the resulting solution was concentrated under reduced pressure and freeze-dried using 1,4-dioxane to quantitatively obtain l-S-n-hutyl 2-acetamido-2-deoxy-3-O-(D-2-propanoyl-L-alanyl-D-isoglutamine)-l-thio-~-D-glucopyranoside (Compound lO) having a melting point of 108 - 112iC
(decomposition).
[~]D5 -15 (c=0.3, methanol) Elemental Analysis for C23H40N4OllS:
Calcd. (~): C, 48.92; H, 7.14; N, 9.92 Found (%)~ C, 48.65; H, 7.49; N, 9.65 --:
Example 9 17 mg of Compound 9 as prepared in Example 7 was dissolved in a mixture of 3 ml of 1,4-dioxane and l ml of methanol, and 2 ml of 0.2M potassium hydroxide was added thereto~ Twenty minutes later, the methyl ester was completely hydrolyzed. The reaction mixture was treated with Amberlite IR-120 (H ), the resulting solution was concentrated and ~reeze-dried from dioxane to quantitatively give l-S hexadecanyl 2-acetamido-2-deoxy-3-O-(D-2-propanoyl-L-alanyl-D-isoglutamine)-l-thio-~-D-glucopyranoside (Compound ll) having a melting point of 110 - 115C (decomposition).
~]D +24 (c=U.15, methanol) Elemental Analysis for C35H64N4OloS:
Calcd. (~): C, 57.35; H, 8.80; N, 7.64 Found (~): C, 57.09; H, 8.95; N, 7.55 Example lO
85 mg of Compound l as prepared in Example 1-(5) was dissolved in 5 ml o~ absolute methanol, and lO mg of sodium methoxide was added thereto. After 1.5 hours, the reaction mixture was treated with Amberlite IRC-50 (~t~ and then concentrated. The resulting syrup was purified by silica gel (lO g) column chromatography (developing agent (a): chloroform; (b): chloro~orm-methanol=40:1). From the effluent (b) was obtained 76 mg ; (yield: 96~) of 2-acetamido-2-deoxy-4,6-O isopropylidene~
. :
3 0-(D-2-propanoyl-L-alanyl-D-isoglutamine methyl ester)-l-thio-~-D-glucopyranose (Compound 12). Recrystallization from diethyl ether gave a product having a melting point of 105 - 107C (decomposition).
[a]D ~33 (c=0.5, chloroform) Elemental Analysis for C23~38N401oS:
Calcd. (%): C, 49.10; H, 6.81; N, 9.96 Found (~): C, 49.Zl; H, 6.77; N, 9.83 Example 11 60 mg of Compound 12 as prepared in Example 10 was dissolved in a mixture of 2 ml of anhydrous dichloro-methane and 1 ml of pyridine, and 1 ml of dichloromethane containing 40 mg of hexa~decanoyl chlorlde was added thereto under cooling. The reaction completed in 1 hour.
lS To the reaction mixture was added 1 ml of methanol, and the mixture was concentrated under reduced pressure.
The resulting syrup was extracted with chloroform, washed with water and dried over sodiurn sulfate. The chloroform was distilled off, and the residue was purified by silica gel -(10 g) column chromatography (developing agent (a): chlO~Qform-methanol=loo:l; (b): chloroform-methanol=
50:1; (c): chloroform-methanol=30:1). From the effluent (c), there was obtained 70 mg (yield: 82~) of 2-acetamido-2-deoxy-1-S-hexadecanQyl-4,6-0-isopropylidene-3-0-(D-2-propanoyl-L-alanyl-D-isoglutamine methyl ester)-l-thio--., ~f~ 3~
~-D-glucopyranose (Compound 13) having a melting point of 98 - 100C.
~a]D +30 ~c=0.2, chloroform) Elemental Analysis for C H N O S:
Calcd. (%): C, 5~8.47; H, 8.56; N, 6.99 Found (%): C, 58.53; H, 8.63; N, 6.75 Example 12 50 mg of Compound 13 as prepared in ~xample 11 was dissolved in 3 ml of a 80% aqueous acetic acid solution and heated at 45C for 1 hour to hydrolyze the isopropylidene group. The reaction mixture was concen-trated under reduced pressure and crystallized from diethyl ether to give 43 mg (yield: 91%) of 2-acetamido~
2-deoxy-1-S-hexadecanoyl-3-O-~D-2-propanoyl-L-alanyl-D-isoglutamine methyl ester)-l-thio-~D-glucopyranose (Compound 14) having a melting point or 181 - 183C which showed a single spot on thin layer chromatography.
[a~D5 '63 (c=0.2, chloroform-methanol~
Elemental Analysis for C36H6~N~OllS:
Calcd. (%): C, 56.82; H, 8.48; N, 7.36 Found (%): C, 56.77; H, 8.56; N, 7.35 Example 13 80 mg of Compound 1 as o~epared in Example 1 (5) was dissolved in 5 ml of absolute methanol, and 10 mg of sodium methoxide was added thereto. After the mixture was allowed to stand a-t room temperature for 1 hour, 5 ml of 0.2M potassium hydroxide was added thereto.
Five minutes later, the base was removed by treating with Amberlite IRC-50 (H ) resin, and the resin was washed with methanol. The filtrate and the washing were combined and concentrated under reduced pressure at a temperature below 40C to obtain 2-acetamido-2-deoxy-~,6-0-isopropylidene-3-G-(D-2-propanoyl ~-alanyl-D-isoglutamine)-l-thio-~-D-glucopyranose (Compound 15). The product was dissolved in a mixture of 2 ml of anhydrous dichloro-methane and l ml of pyridine, and to the resulting solution was added a solution of 45 mg of hexadecanoyl chloride in l ml of anhydrous dichloromethane under ice-cooling, followed by stirring for 1.5 hou~s. The excess of the chloride was decomposed with methanol, and the de-composition solution was concentrated under reduced pres3ure. The residue was extracted with chloroform, washed with water and dried over sodium sulfate.
The chloroform was distilled off, and the resulting syrup was purified by silica yel (20 g) column chromato-graphy (developing agent (a): chloroform-methanol-50:1;
(b) chloroform-methanol=10 1). From the effluent (b), there was obtained 75 mg (yield: 72~) of 2-acetamido-2-deoxy-1-S-hexadecanoyl-4,6~0-isopropylidene-3-0-(D-2-propanoyl-L-alanyl-D-isoglutamine)-l-thio-~-D-glucopyranose (Compound 16) having a melting point of 103 - 106C.
[~]D +42 (c=0.15, chloroform-methanol=l:l) Elemental Analysis for C38H66N4OllS:
Calcd. (%): C, 57.99; H, 8.45; N, 7.12 Found (%) C, 57.65; H, 8.69; N, 7.05 Example 14 30 mg of Compound 16 as prepared in Example 13 was dissolved in 2 ml of a 80% aqueous asetic acid solution and heated at 45C for 1 hour to completely hydrolyze the isopropylidene group. After confirming that the thin layer chromatography showed a single spot, the reaction mixture was freeze-dried to obtain 28 mg of 2-acetamido-2-deoxy-1 S-hexadecanoyl-3-O-(D-2-propanoyl-L-alanyl-D-isoglutamine)-l-thio-~-D-glucopyranose (Compound 17) as a colorless amorphous substance having a melting point of 125 - 129C.
[a] ~ ~62 ~ 5 (caO ~ 2, chloroform-methanol=l:l) Elemental Analysis for C35H62N4OllS:
Calcd. (~): C, 56.28; Ht 8.73; N, 7.50 Found (~): C~ 55.95, H, 8.51; N, 7.46 Example 15 To a solution of 85 mg of Compound 1 in 5 ml of methanol was added 10 mg of sodium methoxide and the mix-ture was kept for 1~ 5 hours at room temperature, and !
- 2~ -treated with Amberlite IRC-50 (H ) resin to remove the base. The mixture was concentrated and the residue was chromatographed on a column of silica gel (10 g) with chloroform and 40:1 chloroform-met~anol. The latter eluate gave 76 mg of 2-acetamido-2-deoxy-4,6-O-isopropylidene-3-O-(D-2-propanoyl-L-alanyl-D-isoglutamine methyl ester)-l-thio-~-D-glucopyranose as crystals having a melting point of 105 - 107C (decomposition).
~a]25 _33o (c-0.5, chloroform) Elemental Analysis for C23H38N4OloS
Calcd. (%): C, 49.10; H, 6.81; N, 9.96 Found . (~): C, 48089; H, 6.85; N, 9.83 To an lce-cooled solution of 80 mg of the above compound in 1 ml of dry pyridine and 2 ml of dichloromethane was dropwise added, with stirring, a solution of 52 mg of decanoyl chloride in 1 ml of dichloromethane, and the mixture wa~ stirred for 40 minutes at 5 to 10. 0.5 ml of methanol was added to the mixture and the resulting mixture was concentrated and then extracted with chloroform.
The extract was washed with water, dried over sodium sulfa~e and concentrated to a syrup which was then chromato-graphed on a column of silica gel (15 g) with (a) 150:1, (b) 70:1, and (c) 30:1 chloroform ~ methanol. Eluant (a) gave 65 mg of amorphous 2-acetamido-2-deoxy-1-S-decanoyl-4,6-O-isopropylidene-3-O-(D-2-propanoyl-L-alanyl-D-isoglutamine methyl ester)-l-thio-~-D-glucopyranose.
[~]D5 +6.6 (c=0.33, chloroform) Elemental Analysis for C33H55N4OllS4 A solution of 50 mg of the above compound in 2 ml of 80 % aqueous acetic acid was heated for 2 hours at 45C and the mixture was concentrated to give a crystalline mass. Recrystallization from ether gave 2-acetamido-2-; deoxy-l-S-decanoyl-3-O-(D-2-propanoyl-L-alanyl-D-iso~lutamine methyl ester)-l-thio-~-D-glucopyranose ~Compound 18) having a melting point of 176.5.
[a]D5 ~41.3 (c=0.44, 1:1 chloroform - methanol).
Elemental AnalysiS ~or C30H52N4llS
Calcd. (%): C, 53.23; H, 7.74; N, 8.28 Found (%)~ C, 53.31; H, 7.89; N, 8.33 Examples 16 and 17 ___ _ ~ ____ The followiny compounds were prepared ln the same manner as above.
2-Acetamido-2-deoxy-1-S-octadecanoyl-3-O-(D-2-propanoyl-L-alanyl-D-isoglutamine methyl ester)-l-thio-~-D glucopyranose (Compound 19) as an amorphous substance~
[~]D5 +29.0 (c=0.64, 1~1 chloroform - methanol) Elemental Analysis for C38H68N4OllS
Calcd. (~): C, 57.84; H, 8.69; N, 7.10 Found (%): C, 57.55; H, 8.73; N, 7.01 2-Acetamido-2-deoxy-1-S-triacontanoyl-3-O-(D-2-propanoyl~L-alanyl-D-isoglutamine methyl ester)-l--thio-~-D-glucopyranose (Compound 20).
m.p. 182, [a]25 +83.3 (c=0.6, l:l'chloroform - methanol).
Elemental Analysis for C50H92N4OllS
Calcd. (%): C, 62.73; H, 9.69; N, 5.85 Found (%): C, 62.65; H, 9.83; N, 5~81 While the invention has been described in detail and with reference to specific embodiments thereof, it will be apparent to one skilled in the art that various changes and modifications can be made therein without departing from the spirit and scope thereof.
for 45 minutes while stirring. The reaction mixture was concentrated under reduced pressure at below 40C. The acetic acid was completely removed as an azeotropic mixture with toluene to obtain 2-acetamido-1-S-acetyl~2-deoxy-3-O-(D-2-propanoyl-L-alanyl-D-isoglutamine methyl ester)-l-thio-~-D-glucopyranose (Compound 2) that showed a single spot on the thin layer chromatogram. ~ecrys~allization from diethyl ether gave a pure product having a melting point of 158 - 162C.
[o~]D5 +62 (c=0.2, chloroform-methanol=l:l) Elemental Analysis for C22H36N4OllS:
Calcd. (%): C, 46.80; H, 6.43; N, 9.92 Found (%): C, 46.51; H, 6.65; N, 9.85 Example 2 20 mg of Compound 2 as prepared in Example 1-(6) was dissolved in 2 ml of absolute methanol, and a small amount of metal sodium was added thereto. Three minutes later, disappearance of any unreacted substances was 10 conEirmed by thin layer chromatography, and the reaction mixture was treated with Amberiite IR-120 (H ~ ollowed by filtration. From the filtrate was obtained 17.6 mg (yield: 95%) of 2-acetamido-2-deoxy-3-O (D-2-propanoyl-L-alanyl-D-isoglutamine methyl ester)-l-thio-~-D-glucopyranose (Compound 3) having a melting point of 117 - 125C
(decomposition).
[a]D5 ~24 (c=0.2, methanol, equilibrium) Elemental Analysis for C20H34N4OloS
Calcd. (g6): C, 45.97; H, 6.56; N, 10.72 20 Found (~): C, 45.49, H, 6.53, N, 10.88 Example 3 25 mg of Compound 2 as prepared in Example 1-(6) was dissolved in 2 ml of absolute methanol, and a small amount of metal sodium was added thereto. Three minutes ,, ~ :
: :, :",:
later, 1 ml of 0.2M potassium hydroxide was added to the reaction mixture. After confirming completion of the reaction by thin layer chromatography, the reaction mixture was treated with Amberlite [R-120 (H ) to remove the base. The solution was concentrated under reduced pressure and freeze-dried from 1,4-dioxane to obtain 22.4 mg (quantitative yield) of 2-acetamido-2-deoxy-3-O-(D-2-propanoyl-L-alanyl-D-isoglutamine)-l-thio-~-D-glucopyranose (Compound 4) having a melting point of 157 -166C (decomposition).
[~]25 -~20 (c=0.2, methanol, e~uilibrium) Elemental Analysis for ClgH32N4OloS:
Calcd. (%): C, 44.87; H, 6.34; N, 11.02 Found (~): C, 44-.45; H, 6.53; N, 10.99 Example 4 60 mg of Compound 1 as prepared in Example 1-(5) was dissolved in 5 ml of absolute methanol, and 6 mg of sodium mekhoxide was added thereto. Ninety minutes later, a~ter it was confirmed by thin layer chromatoyraphy tha-t the reactant had completely be converted to 2-acetamido-l-S-sodium-2-deoxy-4,6-O-isopropylidene-3-O-(D-2-propanoyl-L-alanyl-D~isogulutamine methyl ester)-1-thio-~-D-glucopyranose (Compound 5), 15 mg of n-butyl bromide was added to the reaction mixture, followed by stirring at room temperature overnight. The reaction mixture was concentrated under :
~ .
o~
reduced pressure, and the resulting syrup was purified by silica gel (10 g) column chromatography (developing agent (a): chloroform; (b): chloroform-methanol=50:1) to obtain 60 mg (yield: 98%) of l-S-n-butyl 2 acetamido-2-deoxy-4,6-O-isopropylidene-3-O-(D-2-propanoyl-L-alanyl-D-isoglutamine methyl ester)-l-thio-~-D-glucopyranoside (Compound 6) having a melting point of 75 - 78C.
[a]25 +25 ~c=0.5, methanol) Elemental Analysis for C27H46N4OloS:
Calcd. (%): C, 52.41; H, 7.49; N, 9.06 Found (%): C, 52.25; H, 7.45; N, 9.01 ~ Exampl`e 5 Compound 5 was prepared in the same manner as in Example 4 but using 83 mg of Compound 1. 43 mg of hexadecanyl bromide was added to the reaction mixture containing Compound 5, and the resulting mixture was stirred at room temperature overnight. The reaction mixture was concentrated under reduced pressure and then purified by silica gel (lO g) column chromatography (developing agent (a): chloroform-methanol=100:1, (b): chloroform-methanol=30:1). From the effluent (b)~ there was obtained lO0 mg (yield: 93%) of l-S-hexadecanyl 2-acetamido-2-deoxy-4,6-O-isopropylidene-3-O-(D-2-propanoyl-L-alanyl-D-isoglutamine methyl ester)-l-thio-3-D-glucopyranoside (Compound 7) having a melting ~.2~
point of 93 - 96C.
[a]D +35 (c=0.3, chloroform) Elemental Analysis for C39H70N4OloS
Calcd. (%): C, 59.51; H; 8.97; N, 7.17 Found (%): C, 59.33; H, 8.86; N, 7.15 Example 6 50 mg of Compound 6 as prepared in Example 4 was dissolved in 3 ml of a 8% aqueous acetic acid solution, and hydrolysis was carried out at 45C for 60 minutes.
After confirming by thin layer chromatography, the reaction mixture was freeze-dried to quantitatively obtain l-S-n-butyl 2-acetamido-2-deoxy-3-O-(D-2-propanoyl-L-alanyl-D-isoglutamine methyl ester) l-thio-~-D-glucopyranoside (Compound 8) having a melting point of 155 - 160C
(decomposition).
[a]D ~14 (c=0.3, methanol) Elemental Analysis for C24H42N4OllS:
Calcd. (%): C, 49.81; H, 7.32; N, 9.68 Found (~): C, 49.45; H, 7.61; N, 9.55 Example 7 50 mg of Compound 7 as prepared in Example 5 was dissolved in 3 ml of a 80% aqueous acetic acid solution and heated at 45C for 1 houx. After confirming disappearance of any unreacted substances by thin layer - 23 ~
chromatography, the reaction mixture was freeze-dried to obtain l-S-hexadecanyl 2-acetamido-2-deoxy-3-O-(D-2-propanoyl-L-alanyl-D-isoglutamine methyl ester)-l-thio-~-D-glucopyranoside (Compound 9) having a melting point of 179 - 182C in a quantitative yield.
- [~]D5 +44 (c=0.2, chloroform-methanol=l:l) Elemental Analysis for C36H66N4OloS:
Calcd. (~): C, 57.88; H, 8.91; N, 7.51 Found (%): C, 57.65; H, 8.96; N, 7.43 Example 8 20 mg of Compound 8 as prepa~ed in Example 6 was dissolved in 2 ml of methanol, and 2 ml of 0.2M
potassium hydxoxide was added thereto. Twenty minutes later, the ester was completely hydrolyzed. The reaction ; 15 mixture was treated with Amberlite IR-120 (H ), the resulting solution was concentrated under reduced pressure and freeze-dried using 1,4-dioxane to quantitatively obtain l-S-n-hutyl 2-acetamido-2-deoxy-3-O-(D-2-propanoyl-L-alanyl-D-isoglutamine)-l-thio-~-D-glucopyranoside (Compound lO) having a melting point of 108 - 112iC
(decomposition).
[~]D5 -15 (c=0.3, methanol) Elemental Analysis for C23H40N4OllS:
Calcd. (~): C, 48.92; H, 7.14; N, 9.92 Found (%)~ C, 48.65; H, 7.49; N, 9.65 --:
Example 9 17 mg of Compound 9 as prepared in Example 7 was dissolved in a mixture of 3 ml of 1,4-dioxane and l ml of methanol, and 2 ml of 0.2M potassium hydroxide was added thereto~ Twenty minutes later, the methyl ester was completely hydrolyzed. The reaction mixture was treated with Amberlite IR-120 (H ), the resulting solution was concentrated and ~reeze-dried from dioxane to quantitatively give l-S hexadecanyl 2-acetamido-2-deoxy-3-O-(D-2-propanoyl-L-alanyl-D-isoglutamine)-l-thio-~-D-glucopyranoside (Compound ll) having a melting point of 110 - 115C (decomposition).
~]D +24 (c=U.15, methanol) Elemental Analysis for C35H64N4OloS:
Calcd. (~): C, 57.35; H, 8.80; N, 7.64 Found (~): C, 57.09; H, 8.95; N, 7.55 Example lO
85 mg of Compound l as prepared in Example 1-(5) was dissolved in 5 ml o~ absolute methanol, and lO mg of sodium methoxide was added thereto. After 1.5 hours, the reaction mixture was treated with Amberlite IRC-50 (~t~ and then concentrated. The resulting syrup was purified by silica gel (lO g) column chromatography (developing agent (a): chloroform; (b): chloro~orm-methanol=40:1). From the effluent (b) was obtained 76 mg ; (yield: 96~) of 2-acetamido-2-deoxy-4,6-O isopropylidene~
. :
3 0-(D-2-propanoyl-L-alanyl-D-isoglutamine methyl ester)-l-thio-~-D-glucopyranose (Compound 12). Recrystallization from diethyl ether gave a product having a melting point of 105 - 107C (decomposition).
[a]D ~33 (c=0.5, chloroform) Elemental Analysis for C23~38N401oS:
Calcd. (%): C, 49.10; H, 6.81; N, 9.96 Found (~): C, 49.Zl; H, 6.77; N, 9.83 Example 11 60 mg of Compound 12 as prepared in Example 10 was dissolved in a mixture of 2 ml of anhydrous dichloro-methane and 1 ml of pyridine, and 1 ml of dichloromethane containing 40 mg of hexa~decanoyl chlorlde was added thereto under cooling. The reaction completed in 1 hour.
lS To the reaction mixture was added 1 ml of methanol, and the mixture was concentrated under reduced pressure.
The resulting syrup was extracted with chloroform, washed with water and dried over sodiurn sulfate. The chloroform was distilled off, and the residue was purified by silica gel -(10 g) column chromatography (developing agent (a): chlO~Qform-methanol=loo:l; (b): chloroform-methanol=
50:1; (c): chloroform-methanol=30:1). From the effluent (c), there was obtained 70 mg (yield: 82~) of 2-acetamido-2-deoxy-1-S-hexadecanQyl-4,6-0-isopropylidene-3-0-(D-2-propanoyl-L-alanyl-D-isoglutamine methyl ester)-l-thio--., ~f~ 3~
~-D-glucopyranose (Compound 13) having a melting point of 98 - 100C.
~a]D +30 ~c=0.2, chloroform) Elemental Analysis for C H N O S:
Calcd. (%): C, 5~8.47; H, 8.56; N, 6.99 Found (%): C, 58.53; H, 8.63; N, 6.75 Example 12 50 mg of Compound 13 as prepared in ~xample 11 was dissolved in 3 ml of a 80% aqueous acetic acid solution and heated at 45C for 1 hour to hydrolyze the isopropylidene group. The reaction mixture was concen-trated under reduced pressure and crystallized from diethyl ether to give 43 mg (yield: 91%) of 2-acetamido~
2-deoxy-1-S-hexadecanoyl-3-O-~D-2-propanoyl-L-alanyl-D-isoglutamine methyl ester)-l-thio-~D-glucopyranose (Compound 14) having a melting point or 181 - 183C which showed a single spot on thin layer chromatography.
[a~D5 '63 (c=0.2, chloroform-methanol~
Elemental Analysis for C36H6~N~OllS:
Calcd. (%): C, 56.82; H, 8.48; N, 7.36 Found (%): C, 56.77; H, 8.56; N, 7.35 Example 13 80 mg of Compound 1 as o~epared in Example 1 (5) was dissolved in 5 ml of absolute methanol, and 10 mg of sodium methoxide was added thereto. After the mixture was allowed to stand a-t room temperature for 1 hour, 5 ml of 0.2M potassium hydroxide was added thereto.
Five minutes later, the base was removed by treating with Amberlite IRC-50 (H ) resin, and the resin was washed with methanol. The filtrate and the washing were combined and concentrated under reduced pressure at a temperature below 40C to obtain 2-acetamido-2-deoxy-~,6-0-isopropylidene-3-G-(D-2-propanoyl ~-alanyl-D-isoglutamine)-l-thio-~-D-glucopyranose (Compound 15). The product was dissolved in a mixture of 2 ml of anhydrous dichloro-methane and l ml of pyridine, and to the resulting solution was added a solution of 45 mg of hexadecanoyl chloride in l ml of anhydrous dichloromethane under ice-cooling, followed by stirring for 1.5 hou~s. The excess of the chloride was decomposed with methanol, and the de-composition solution was concentrated under reduced pres3ure. The residue was extracted with chloroform, washed with water and dried over sodium sulfate.
The chloroform was distilled off, and the resulting syrup was purified by silica yel (20 g) column chromato-graphy (developing agent (a): chloroform-methanol-50:1;
(b) chloroform-methanol=10 1). From the effluent (b), there was obtained 75 mg (yield: 72~) of 2-acetamido-2-deoxy-1-S-hexadecanoyl-4,6~0-isopropylidene-3-0-(D-2-propanoyl-L-alanyl-D-isoglutamine)-l-thio-~-D-glucopyranose (Compound 16) having a melting point of 103 - 106C.
[~]D +42 (c=0.15, chloroform-methanol=l:l) Elemental Analysis for C38H66N4OllS:
Calcd. (%): C, 57.99; H, 8.45; N, 7.12 Found (%) C, 57.65; H, 8.69; N, 7.05 Example 14 30 mg of Compound 16 as prepared in Example 13 was dissolved in 2 ml of a 80% aqueous asetic acid solution and heated at 45C for 1 hour to completely hydrolyze the isopropylidene group. After confirming that the thin layer chromatography showed a single spot, the reaction mixture was freeze-dried to obtain 28 mg of 2-acetamido-2-deoxy-1 S-hexadecanoyl-3-O-(D-2-propanoyl-L-alanyl-D-isoglutamine)-l-thio-~-D-glucopyranose (Compound 17) as a colorless amorphous substance having a melting point of 125 - 129C.
[a] ~ ~62 ~ 5 (caO ~ 2, chloroform-methanol=l:l) Elemental Analysis for C35H62N4OllS:
Calcd. (~): C, 56.28; Ht 8.73; N, 7.50 Found (~): C~ 55.95, H, 8.51; N, 7.46 Example 15 To a solution of 85 mg of Compound 1 in 5 ml of methanol was added 10 mg of sodium methoxide and the mix-ture was kept for 1~ 5 hours at room temperature, and !
- 2~ -treated with Amberlite IRC-50 (H ) resin to remove the base. The mixture was concentrated and the residue was chromatographed on a column of silica gel (10 g) with chloroform and 40:1 chloroform-met~anol. The latter eluate gave 76 mg of 2-acetamido-2-deoxy-4,6-O-isopropylidene-3-O-(D-2-propanoyl-L-alanyl-D-isoglutamine methyl ester)-l-thio-~-D-glucopyranose as crystals having a melting point of 105 - 107C (decomposition).
~a]25 _33o (c-0.5, chloroform) Elemental Analysis for C23H38N4OloS
Calcd. (%): C, 49.10; H, 6.81; N, 9.96 Found . (~): C, 48089; H, 6.85; N, 9.83 To an lce-cooled solution of 80 mg of the above compound in 1 ml of dry pyridine and 2 ml of dichloromethane was dropwise added, with stirring, a solution of 52 mg of decanoyl chloride in 1 ml of dichloromethane, and the mixture wa~ stirred for 40 minutes at 5 to 10. 0.5 ml of methanol was added to the mixture and the resulting mixture was concentrated and then extracted with chloroform.
The extract was washed with water, dried over sodium sulfa~e and concentrated to a syrup which was then chromato-graphed on a column of silica gel (15 g) with (a) 150:1, (b) 70:1, and (c) 30:1 chloroform ~ methanol. Eluant (a) gave 65 mg of amorphous 2-acetamido-2-deoxy-1-S-decanoyl-4,6-O-isopropylidene-3-O-(D-2-propanoyl-L-alanyl-D-isoglutamine methyl ester)-l-thio-~-D-glucopyranose.
[~]D5 +6.6 (c=0.33, chloroform) Elemental Analysis for C33H55N4OllS4 A solution of 50 mg of the above compound in 2 ml of 80 % aqueous acetic acid was heated for 2 hours at 45C and the mixture was concentrated to give a crystalline mass. Recrystallization from ether gave 2-acetamido-2-; deoxy-l-S-decanoyl-3-O-(D-2-propanoyl-L-alanyl-D-iso~lutamine methyl ester)-l-thio-~-D-glucopyranose ~Compound 18) having a melting point of 176.5.
[a]D5 ~41.3 (c=0.44, 1:1 chloroform - methanol).
Elemental AnalysiS ~or C30H52N4llS
Calcd. (%): C, 53.23; H, 7.74; N, 8.28 Found (%)~ C, 53.31; H, 7.89; N, 8.33 Examples 16 and 17 ___ _ ~ ____ The followiny compounds were prepared ln the same manner as above.
2-Acetamido-2-deoxy-1-S-octadecanoyl-3-O-(D-2-propanoyl-L-alanyl-D-isoglutamine methyl ester)-l-thio-~-D glucopyranose (Compound 19) as an amorphous substance~
[~]D5 +29.0 (c=0.64, 1~1 chloroform - methanol) Elemental Analysis for C38H68N4OllS
Calcd. (~): C, 57.84; H, 8.69; N, 7.10 Found (%): C, 57.55; H, 8.73; N, 7.01 2-Acetamido-2-deoxy-1-S-triacontanoyl-3-O-(D-2-propanoyl~L-alanyl-D-isoglutamine methyl ester)-l--thio-~-D-glucopyranose (Compound 20).
m.p. 182, [a]25 +83.3 (c=0.6, l:l'chloroform - methanol).
Elemental Analysis for C50H92N4OllS
Calcd. (%): C, 62.73; H, 9.69; N, 5.85 Found (%): C, 62.65; H, 9.83; N, 5~81 While the invention has been described in detail and with reference to specific embodiments thereof, it will be apparent to one skilled in the art that various changes and modifications can be made therein without departing from the spirit and scope thereof.
Claims (33)
1. A process for the preparation of a muramyldipeptide derivative represented by the formula (I):
wherein Ala represents alanine; Acyl represents an acyl group having 2 to 6 carbon atoms; R1 and R2 each represents a hydro-gen atom or, when taken together, may form an alkylidene group having 1 to 6 carbon atoms; R3 represents a hydrogen atom, an acyl group having 2 to 40 carbon atoms or an alkyl group having 1 to 40 carbon atoms; and R4 represents a hydrogen atom or an alkyl group having 1 to 6 carbon atoms, comprising a process selected from the group consisting of:
(a) a process to produce compounds of the formula (I) wherein R1 and R2 when taken together form an alkylidene group having 1 to 6 carbon atoms, R3 is an acyl group having 2 to 40 carbon atoms and R4 is an alkyl group having 1 to 6 carbon atoms, comprising condensing a compound of the formula IV
IV
wherein R31 represents an acyl group having 2 to 40 carbon atoms, R5 represents a hydrogen atom or an alkyl group having 1 to 6 carbon atoms, and R6 represents an alkyl group having 1 to 6 carbon atoms, with an alanyl-isoglutamine alkyl ester, (b) a process to produce compounds of the formula (I) wherein R1 and R2 when taken together form an alkylidene group having 1 to 6 carbon atoms, R3 is a hydrogen atom, and R4 is an alkyl group having 1 to 6 carbon atoms, com-prising process (a) above further including reacting the resultant product of process (a) with. a metal alcoholate;
(c) a process to produce compounds of the formula (I) wherein R1 and R2 when taken together form an alkylidene group having 1 to 6 carbon atoms, R3 is an alkyl group having 1 to 40 carbon atoms, and R4 is an alkyl group having 1 to 6 carbon atoms, comprising process (b) above further including reacting the resultant product of process (b) with an alkyl halide;
(d) a process to produce compounds of the formula (I) wherein R1 and R2 each is a hydrogen atom, R3 is an acyl group having 2 to 40 carbon atoms and R4 is an alkyl group having 1 to 6 carbon atoms, comprising process (a) above further including the step of cleavage of the alkylidene group;
(e) a process to produce compounds of the formula (I) wherein R1 and R2 when taken together from an alkylidene group having 1 to 6 carbon atoms, R3 is an acyl group having 2 to 40 carbon atoms, and R4 is a hydrogen atom, comprising process (a) above further including the step of hydrolysis of the ester group;
(f) a process to produce compounds of the formula (I) wherein R1 and R2 each is a hydrogen atom, R3 is an acyl group having 2 to 40 carbon atoms, and R4 is a hydrogen atom, comprising process (a) above further including the steps of cleavage of the alkylidene group and hydrolysis of the ester group;
(g) a process to produce compounds of the formula (I) wherein R1 and R2 each is a hydrogen atom, R3 is a hydrogen atom, and R4 is an alkyl group having 1 to 6 carbon atoms comprising process (b) above further including the step of cleavage of the alkylidene group;
(h) a process to produce compounds of the formula [I) wherein R1 and R2 when taken together form an alkylidene group having 1 to 6 carbon atoms, R3 is a hydrogen atom, and R4 is a hydrogen atom, comprising process (b) above further including the step of hydrolysis of the ester group;
(i) a process to produce compounds of the formula (I) wherein R1, R2, R3 and R4 each is a hydrogen atom comprising process (b) above further including the steps of cleavage of the alkylidene group and hydrolysis of the ester group;
(j) a process to produce compounds of the formula (I) wherein R1 and R2 each is a hydrogen atom, R3 is an alkyl group having 1 to 40 carbon atoms, and R4 is an alkyl group having 1 to 6 carbon atoms, comprising process (c) above further including the step of cleavage of the alkylidene group;
(k) a process to produce compounds of the formula (I) wherein R1 and R2 when taken together form an alkylidene group having 1 to 6 carbon atoms, R3 is an alkyl group having 1 to 40 carbon atoms, and R4 is a hydrogen atom, comprising process (c) above further including the step of hydrolysis of the ester group, and (l) a process to produce compounds of the formula (I) wherein R1, R2 and R4 each is a hydrogen atom and R3 is an alkyl group having 1 to 40 carbon atoms, comprising process (c) above further including the steps of cleavage of the alkylidene group and hydrolysis of the ester group.
wherein Ala represents alanine; Acyl represents an acyl group having 2 to 6 carbon atoms; R1 and R2 each represents a hydro-gen atom or, when taken together, may form an alkylidene group having 1 to 6 carbon atoms; R3 represents a hydrogen atom, an acyl group having 2 to 40 carbon atoms or an alkyl group having 1 to 40 carbon atoms; and R4 represents a hydrogen atom or an alkyl group having 1 to 6 carbon atoms, comprising a process selected from the group consisting of:
(a) a process to produce compounds of the formula (I) wherein R1 and R2 when taken together form an alkylidene group having 1 to 6 carbon atoms, R3 is an acyl group having 2 to 40 carbon atoms and R4 is an alkyl group having 1 to 6 carbon atoms, comprising condensing a compound of the formula IV
IV
wherein R31 represents an acyl group having 2 to 40 carbon atoms, R5 represents a hydrogen atom or an alkyl group having 1 to 6 carbon atoms, and R6 represents an alkyl group having 1 to 6 carbon atoms, with an alanyl-isoglutamine alkyl ester, (b) a process to produce compounds of the formula (I) wherein R1 and R2 when taken together form an alkylidene group having 1 to 6 carbon atoms, R3 is a hydrogen atom, and R4 is an alkyl group having 1 to 6 carbon atoms, com-prising process (a) above further including reacting the resultant product of process (a) with. a metal alcoholate;
(c) a process to produce compounds of the formula (I) wherein R1 and R2 when taken together form an alkylidene group having 1 to 6 carbon atoms, R3 is an alkyl group having 1 to 40 carbon atoms, and R4 is an alkyl group having 1 to 6 carbon atoms, comprising process (b) above further including reacting the resultant product of process (b) with an alkyl halide;
(d) a process to produce compounds of the formula (I) wherein R1 and R2 each is a hydrogen atom, R3 is an acyl group having 2 to 40 carbon atoms and R4 is an alkyl group having 1 to 6 carbon atoms, comprising process (a) above further including the step of cleavage of the alkylidene group;
(e) a process to produce compounds of the formula (I) wherein R1 and R2 when taken together from an alkylidene group having 1 to 6 carbon atoms, R3 is an acyl group having 2 to 40 carbon atoms, and R4 is a hydrogen atom, comprising process (a) above further including the step of hydrolysis of the ester group;
(f) a process to produce compounds of the formula (I) wherein R1 and R2 each is a hydrogen atom, R3 is an acyl group having 2 to 40 carbon atoms, and R4 is a hydrogen atom, comprising process (a) above further including the steps of cleavage of the alkylidene group and hydrolysis of the ester group;
(g) a process to produce compounds of the formula (I) wherein R1 and R2 each is a hydrogen atom, R3 is a hydrogen atom, and R4 is an alkyl group having 1 to 6 carbon atoms comprising process (b) above further including the step of cleavage of the alkylidene group;
(h) a process to produce compounds of the formula [I) wherein R1 and R2 when taken together form an alkylidene group having 1 to 6 carbon atoms, R3 is a hydrogen atom, and R4 is a hydrogen atom, comprising process (b) above further including the step of hydrolysis of the ester group;
(i) a process to produce compounds of the formula (I) wherein R1, R2, R3 and R4 each is a hydrogen atom comprising process (b) above further including the steps of cleavage of the alkylidene group and hydrolysis of the ester group;
(j) a process to produce compounds of the formula (I) wherein R1 and R2 each is a hydrogen atom, R3 is an alkyl group having 1 to 40 carbon atoms, and R4 is an alkyl group having 1 to 6 carbon atoms, comprising process (c) above further including the step of cleavage of the alkylidene group;
(k) a process to produce compounds of the formula (I) wherein R1 and R2 when taken together form an alkylidene group having 1 to 6 carbon atoms, R3 is an alkyl group having 1 to 40 carbon atoms, and R4 is a hydrogen atom, comprising process (c) above further including the step of hydrolysis of the ester group, and (l) a process to produce compounds of the formula (I) wherein R1, R2 and R4 each is a hydrogen atom and R3 is an alkyl group having 1 to 40 carbon atoms, comprising process (c) above further including the steps of cleavage of the alkylidene group and hydrolysis of the ester group.
2. A muramyldipeptide derivative represented by the formula:
I
wherein Ala represents alanine; Acyl represents an acyl group having 2 to 6 carbon atoms; R1 and R2 each represents a hydrogen atom or, when taken together, may form an alkylidene group having 1 to 6 carbon atoms; R3 represents a hydrogen atom, an acyl group having 2 to 40 carbon atoms or an alkyl group having 1 to 40 carbon atoms; and R3 represents a hydro-gen atom or an alkyl group having 1 to 6 carbon atoms, when produced by the process of claim 1 or an obvious chemi-cal equivalent.
I
wherein Ala represents alanine; Acyl represents an acyl group having 2 to 6 carbon atoms; R1 and R2 each represents a hydrogen atom or, when taken together, may form an alkylidene group having 1 to 6 carbon atoms; R3 represents a hydrogen atom, an acyl group having 2 to 40 carbon atoms or an alkyl group having 1 to 40 carbon atoms; and R3 represents a hydro-gen atom or an alkyl group having 1 to 6 carbon atoms, when produced by the process of claim 1 or an obvious chemi-cal equivalent.
3. Process (a) of claim 1.
4. The compound of formula (I) as claimed in claim 2 wherein R1 and R2 when taken together form an alkylidene group having 1 to 6 carbon atoms, R3 is an acyl group having 2 to 40 carbon atoms and R4 is an alkyl group having 1 to 6 carbon atoms when produced by the process of claim 3 or an obvious chemical equivalent.
5. Process (b) of claim 1.
6. Process (c) of claim 1.
7. Process (d) of claim 1.
8. The compound of the formula (I) as claimed in claim 2 wherein R1 and R2 each is a hydrogen atom, R3 is an acyl group having 2 to 40 carbon atoms and R4 is an alkyl group having 1 to 6 carbon atoms when produced by the process of claim 7.
9. Process (e) of claim 1.
10. Process (f) of claim 1.
11. Process (g) of claim 1.
12. Process (h) of claim 1.
13. Process (i) of claim 1.
14. Process (j) of claim 1.
15. Process (k) of claim 1.
16. Process (l) of claim 1,
17. A process to produce 2-acetamido-2-deoxy-l-S-decanoyl-3-0-(D-2-propanoyl-L-alanyl-D-isoglutamine methyl ester)-l-thio-.beta.-D-glucopyranose comprising condensing 2-acetamido-2-deoxy-l-S-decanoyl-4,6-0-isopropylidene-3-0-(D-2-propanoyl)-l-thio-.beta.-D-glucopyranose with alanyl-iso-glutamine methyl ester and subjecting the resultant product to cleavage of its alkylidene group.
18. 2-Acetamido-2-deoxy-l-S-decanoyl-3-0-(D-2-propanoyl-L-alanyl-D-isoglutamine methyl ester)-l-thio-.beta.-D-glucopyranose, when prepared by the process of claim 17 on an obvious chemical equivalent.
19. A process to produce 2-acetamido-l-S-acetyl-2-deoxy-3-0-(D-2-propanoyl-L-alanyl-D-isoglutamine methyl ester)-l-thio-.beta.-D-glucopyranose, comprising condensing 2-acetamido-2-deoxy-l-S-acetyl-4,6-0-isopropylidene-3-O-(D-2 propanoyl)-l-thio-.beta.-D-glucopyranose with alanyl-isoglutamine methyl ester and subjecting the resultant product to cleavage of its al-kylidene group.
20. 2-Acetamido-l-S-acetyl-2-deoxy-3-O-(D-2-propanoyl-L-alanyl-D-isoglutamine methyl ester)-l-thio-.beta.-D-glucopyranose, when prepared by the process of claim 19 or an obvious chemical equivalent.
21. A process to prepare 2-acetamido-2-deoxy-3-O-(D-2-propanoyl-L-alanyl-D-isoglutamine methyl ester)-l-thio-.beta.-D-glucopyranose, comprising condensing 2-acetamido-2-deoxy-l-S-acetyl-4,6-0-isopropylidene-3-O-(D-2-propanoyl)-l-thio-.beta.-D-glucopyranose with alanyl-isoglutamine methyl ester and then subjecting the resultant product to reaction with a metal al-coholate and to cleavage of its alkylidene group.
22. 2-Acetamido-2-deoxy-3-O-(D-2-propanoyl-L-alanyl-D-isoglutamine methyl ester)-l-thio-.beta.-D-glucopyranose, when prepared by the process of claim 21 or an obvious chemical equivalent.
23. A process to prepare 2-acetamido-2-deoxy-3-O-(D-2-propanoyl-L-alanyl-D-isoglutamine)-1-thiO-.beta.-D-glucopyra-nose comprising condensing 2-acetamido-2-deoxy-1-S-acetyl-4, 6-O-isopropylidene-3-O-(D-2-propanoyl)-1-thio-.beta.-D-glucopyranose with alanyl-isoglutamine methyl ester, and then subjecting the resultant product to reaction with a metal alcoholate, to cleavage of its alkylidene group and to hydrolysis of its ester group.
24. 2-Acetamido-2-deoxy-3-O-(D-2-propanoyl-L-alanyl-D-isoglutamine)-l-thio-.beta.-D-glucopyranose, when prepared by the process of claim 23 or an obvious chemical equivalent.
25. A process to prepare 2-acetamido-2-deoxy-l-S-hexadecanoyl-3-O-(D-2-propanoyl-L-alanyl-D-isoglutamine methyl ester)-l-thio-.beta.-D-glucopyranose, comprising condensing 2-acetamido-2-deoxy-l-S-hexadecanoyl-4,6-O-isopropylidene-3-O-(D-2-propanoyl)-l-thio-.beta.-D-glucopyranose with alanyl-isoglutamine methyl ester and subjecting the resultant product to cleavage of its alkylidene group.
26. 2-Acetamido-2-deoxy-l-S-hexadecanoyl-3-O-(D-2-propanoyl-L-alanyl-D-isoglutamine methyl ester)-l-thio-.beta.-D-glucopyranose, when prepared by the process of claim 25 or an obvious chemical equivalent.
27. A process to prepare 2-acetamido-2-deoxy-l-S-hexa-decanoyl-3-O-(D-2-propanoyl-L-alanyl-D-isoglutamine)-l-thio-.beta.-D glucopyranose, comprising the process of claim 25 further including subecting the resultant product to hydrolysis of its ester group.
28. 2-Acetamido-2-deoxy-1-S-hexadecanoyl-3-O-(D-2-propanoyl-L-alanyl-D-isoglutamine)-1-thio-.beta.-D-glucopyranose, when prepared by the process of claim 27 or an obvious chemi-cal equivalent.
29. A process to prepare 2-acetamido-2-deoxy-1-S-eicosanoyl-3-O-(D-2-propanoyl-L-alanyl-D-isoglutamine)-1-thio-.beta.-D-glucopyranose, comprising condensing 2-acetamino-2-deoxy-1-S-eicosanoyl-4,6-O-isopropylidene-3-O-(D-2-propanoyl) -1-thio-.beta.-D-glucopyranose with alanyl-isoglutamine alkyl ester and subjecting the resultant product to cleavage of its alkylidene group and then to hydrolysis of its ester group.
30. 2-Acetamido-2-deoxy-1-S-eicosanoyl-3-O-(D-2-propanoyl-L-alanyl-D-isoglutamine)-1-thio-.beta.-D-glucopyranose, when prepared by the process of claims 29 or an obvious chemical equivalent.
31. The process claimed in claim 1 wherein R3 represents an acyl group having 2 to 6 carbon atoms or an alkyl group having 1 to 40 carbon atoms.
32. The process claimed in claim 3, 5 or 6 wherein R3 represents an acyl group having 2 to 5 carbon atoms or an alkyl group having 1 to 40 carbon atoms.
33. The process claimed in claim 1 wherein R3 represents an acyl group having 2 to 6 carbon atoms or an alkyl group having 1 to 6 carbon atoms.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP7051982A JPH0248671B2 (en) | 1982-04-28 | 1982-04-28 | KASADAKADEKEIRYONAGOSEIHIKAKU |
| JP130705/1982 | 1982-07-27 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CA1249400A true CA1249400A (en) | 1989-01-24 |
Family
ID=13433851
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA000432987A Expired CA1249400A (en) | 1982-04-28 | 1983-07-22 | Muramyldipeptide derivatives |
Country Status (2)
| Country | Link |
|---|---|
| JP (1) | JPH0248671B2 (en) |
| CA (1) | CA1249400A (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH0694627B2 (en) * | 1989-04-19 | 1994-11-24 | 帝人コードレ株式会社 | Leather-like sheet |
-
1982
- 1982-04-28 JP JP7051982A patent/JPH0248671B2/en not_active Expired - Lifetime
-
1983
- 1983-07-22 CA CA000432987A patent/CA1249400A/en not_active Expired
Also Published As
| Publication number | Publication date |
|---|---|
| JPH0248671B2 (en) | 1990-10-25 |
| JPS58191282A (en) | 1983-11-08 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US4101536A (en) | Muramyldipeptide derivatives and process for the preparation thereof | |
| US4331803A (en) | Novel erythromycin compounds | |
| US4317771A (en) | Muramyldipeptide derivatives | |
| KR900016242A (en) | N-acyl derivatives of LL-E33288, antitumor antibiotics and methods for their production and use thereof | |
| CS205025B2 (en) | Process for preparing derivatives of glukosamine | |
| US4606857A (en) | Muramyldipeptide derivatives | |
| JPH0371440B2 (en) | ||
| CA1249400A (en) | Muramyldipeptide derivatives | |
| CA1298579C (en) | Glycolipid containing n-glycolylneuraminic acid and method of producingthe same | |
| SU656505A3 (en) | Method of obtaining a-3-(3,4-dioxyphenyl)-2-methylallaninepeptides | |
| CA1185236A (en) | Immunologically active dipeptidyl saccharides and methods of preparation | |
| US3869347A (en) | Process for the production of new pepstatins having anti-pepsin activity | |
| Yamamoto et al. | Studies on Amino Sugars. I. Preparation of N-Acyl Derivatives of 2-Acetamido-2-deoxy-β-D-glucosylamine | |
| FR2458556A1 (en) | PAROMOMYCIN DERIVATIVES, PROCESS FOR PREPARING THEM AND THEIR USE AS A MEDICINAL PRODUCT | |
| CA1335892C (en) | Saccharide derivatives | |
| Knapp Jr | Telluroamino acids: synthesis of telluromethionine | |
| US4515715A (en) | L-Alanyl-D-isoglutamine adamantylamide | |
| EP0222172B1 (en) | N-glycolylneuraminic acid derivative | |
| US3423398A (en) | Sangivamycin and derivatives thereof | |
| Seto et al. | Isolation and Structure of Urochloralic Acid from Urine of Calves Fed Trichloroethylene1 | |
| IL39302A (en) | Antibiotic em-49 | |
| US3950325A (en) | Novel process for preparing 2,2'-cyclocytidine or 1-β-D-arabinofuranosyl cytosine | |
| US3975366A (en) | Process for the production of new pepstatins having anti-pepsin activity | |
| Tamura et al. | Actinotiocin, a new sulfur-containing peptide antibiotic from Actinomadura pusilla | |
| RU2024546C1 (en) | β-GLYCYRRHIZIC ACID GLYCOPEPTIDE WITH L-GLUTAMIC ACID DIBUTYL ETHER SHOWING ANTI-AIDS-ACTIVITY |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| MKEX | Expiry |