JPS6345280A - Ikarugamycin derivative - Google Patents

Abstract

NEW MATERIAL:The compound of formula I {A is group of formula II (R1 is H or CH3) or group of formula III [R2 is R1; R3 is CH3, (substituted) allyl and benzyl or phenacyl]}. USE:A drug useful as a preventive, remedy and treatment agent for infectious diseases of human and animal with Gram-positive bacteria, fungi and mycoplasma, a feed additive for animal or a preventive or elimination agent for contamination with mycoplasma in cell culture. PREPARATION:A compound of formula I wherein A is group of formula II and R1 is CH3 can be produced by methylating ikarugamycin of formula IV with dimethyl sulfate in the presence of an alkali metal carbonate in a solvent such as dimethylformamide preferably at 20-50 deg.C for 3-15hr. The product can be converted to another compound of formula I wherein A is group of formula II and R1 is H by reacting at 0-80 deg.C for 5min-5hr in the presence of pyridine, thereby demethylating the compound.

Description

DETAILED DESCRIPTION OF THE INVENTION [Industrial Field of Application] The present invention relates to a novel icargamison derivative, and more specifically to the following general formula (1).

(represents a hydrogen atom or a methyl group) or the group represents a methyl radical, R1 represents a methyl group, an allyl group that may have a substituent group, a beazyl group or a zener group that may have a substituent group] The present invention relates to the represented icargamycin derivatives.

[Conventional technology and its problems]

Mycoplasma is not only pathogenic to humans and animals, but also contaminates the culture medium during tissue culture and suppresses cell growth.

It is a microorganism that is known to suppress the production of substances from cells and even cause cell death. Therefore.

Research and development of effective preventive and therapeutic drugs for mycoplasma, which is pathogenic to humans and animals (mainly livestock and poultry), has been carried out, but it is difficult to say that the results are sufficient, and there is a need for even better compounds. development is desired. In addition, many of the mycoplasmas that are contaminated during tissue culture are not pathogenic to humans or animals, and have not been the subject of research until now, which is why no excellent antibacterial agents for tissue culture media have been developed. It was the current situation.

[Means to solve the problem]

The present inventors conducted various studies to obtain a substance with excellent anti-mycoplasma action, and found that a new compound partially modified with Ikarugamysino has excellent anti-mycoplasma action. The present invention has been completed based on this discovery.

Therefore, the present invention provides a novel icargamainone derivative represented by the above formula (1).

The ikarugamay/in derivative (1) of the present invention further comprises the following two groups of compounds (Ia) and (Ib).

(In the formula, R, R, and It have the meanings described above.) Each of them is prepared by, for example, one of the methods shown below.

Method 1: For compounds of formula (I a), RI is a methyl group.

Ikarugamaiso 7 (n) Manufactured by subjecting it to an e-methylation reaction. The starting material, icargamycin, is a compound represented by the following formula (It), which has anti-trichomoniacal activity as well as antibacterial activity against Durham-positive bacteria (Japanese Patent Publication No. 46-28833 and Journal of Antibiotinox vol, 25i5127
1-280.1972).

The above methylation reaction was carried out according to a conventional method using dimethyl sulfuric acid and an alkali carbonate, and dimethyl formamide.

Preferably, the reaction is carried out in a solvent such as dimethyl sulfoxide at a temperature of 20 to 50°C for 3 to 15 hours.

Method 2 A compound in formula (la) in which R1 is a hydrogen atom is produced by subjecting the corresponding compound in which R5 is a methyl group to a demethylation reaction. This demethylation reaction proceeds quantitatively by leaving it for 5 minutes to 5 hours at a temperature of 0 to 80° C. in the presence of a tertiary amine such as pyridino, and methoxyl groups are demethylated.

Method 3: The compound represented by formula (Ib) is prepared from Ikarugamai 7 (
n) and methyl halide, substituent? Allyl halide that may have a substituent, benzyl halide that may have a substituent, or phenacyl halide in a medium such as dimethylformamide or dimethyl sulfoxide in the presence of an alkali carbonate at a temperature of ~80°C for 1 to 5 hours. Manufactured by stirring.

The halogen atom of the halogen 1t'' product used in this reaction is preferably chlorine, bromine or iodine, and specific examples of allyl-ride which may have a substituent include allyl bromide, l-bromo-3methyl Examples of the benzyl halide in which -2-butene etc. can have a substituent include benzyl bromide, p-methoxybenzyl bromide, p-bromobenzyl bromide, etc.; examples of the phenacyl halide include phenacyl bumide, p- Examples include bromphenacil bromide.

Next, regarding the thus obtained icargamycin derivative (1) of the present invention, what is the antibacterial activity? The results of the investigation are shown in Tables 1 and 2. As is clear from these results, the compound (1) of the present invention exhibited a certain level of antibacterial activity against pathogenic microorganisms such as various Durum-positive bacteria, fungi, and mycoplasma, and exhibited particularly excellent activity against mycoplasma. .

(1) Antibacterial spectrum test method> Bacteria and fungi were measured according to the MIC measurement method of the Japanese Society of Chemotherapy. The media used were Mueller Hinoton Medilum (manufactured by Difco) for bacteria and glucose peptone agar medium for fungi. Also, regarding mycoplasma, what is the test mycoplasma? The cells were cultured in Chanonk's liquid medium, and the minimum inhibitory concentration of gold was determined by the liquid dilution method using the same medium.

0.002% phenol red i was added to the medium, and the judgment was positive.Is there a color change in Conotrol? I did it on the day I woke up.

As is clear from Table 2 below, the compound (1) of the present invention was found to have excellent antibacterial activity against mycoplasma. Therefore, the compound (1) of the present invention is suitable for both humans and animals (
For example, it can be used for the prevention, treatment, and treatment of mycoplasma infections in mammals, poultry, etc.).

When the compound (1) of the present invention is used as a drug for the prevention, treatment, or treatment of infectious diseases, the compound may be formulated as it is according to a conventional method.

For formulation, a variety of commonly used media are used, including water, salt solutions, alcohol, and vegetable oils.

Polyethylene glycol, gelatin, lactose.

Amylose, magnesium stearinoate, talc.

Paraffin, aromatic oil, fatty acid monoglyceride, dicryceride, droxymethyl cellulose, etc. can be used, and the dosage form can be capsules, single tablets, powders, *7O tablets, suppositories, injections, etc. In addition, it can be used directly as a feed additive or mixed into feed as a premix shaving for the purpose of preventing infection and promoting the growth of animals.

On the other hand, recently, research on culturing cells and on producing useful substances by culturing whole cells in large quantities has been actively conducted. However, I! Mycoplasma contamination is a problem when culturing II cells, and due to this contamination!
It has been pointed out that it affects the proliferation and substance production of ll cells. Methods for preventing such contamination and removing mycoplasma include antibiotic treatment, high temperature treatment,
Treatment with specific antiserum, treatment with chemicals, etc. are listed, but
Both have their advantages and disadvantages, and generally they are often treated with antibiotics. Since the compound (1) of the present invention exhibits a good action against mycoplasma, it can be expected to have an excellent action in contaminating, preventing and removing mycoplasma.

〔Effect of the invention〕

The Ikarugamai 7 derivative (1) of the present invention has an antibacterial effect on Durum-positive bacteria, fungi, and mycoplasma, and therefore can be used for the prevention and treatment of Durum-positive bacteria, fungi, and mycoplasma infections in humans and animals.

It is useful as a control agent or as an animal feed additive. It is also useful as a preventive or removal agent for mycoplasma contamination in cell culture.

[Implementation row]

Next, the present invention will be explained in detail with reference to Examples, but the present invention is not limited to these Examples.

Example 1 Icargamycin 0.96 g (2 mmol) dimethylformamide 60 ml #a'/1 & potassium carbonate 5
52,! i' (40 ml mol) f In addition, 3.86 g (30 mmol) dimethyl sulfate was added under stirring at room temperature.
f It was added dropwise over about 1 hour, and further stirred under the same conditions for 10 hours. After pressurizing the reaction 'ttk into 200 ml of ice water and leaving it at 4°C overnight.

All precipitated precipitate was collected and dried. This precipitate was separated and purified by silica gel column C mudgraphy, and 2% (
V/V) The crystals obtained from the methanol-chloroform mixture elution fraction were recrystallized from an acetone-methanol mixture to obtain (Ia) colorless needle-like crystals of the icargamycin derivative (compound l) represented by R= -CH3 in the formula. 0.32
g (yield 40.o) was obtained.

Melting point 169-171℃ UV λ”t0H311nrn 'H-NMRδppm (CDC13) 7.35(d
, LH), 6.44 (d, d, IH), 4.
32(s, 3H).

3.00 (s, 3H) 13C-NMRδ ppm (CDCis) 194.
3(s), 182.1(s), 178.6(s), 1
66.5(s).

146.0(d), 139.2(d), 1310(d)
), 129.8(d).

128.3(d), 124.3(d), 100.7(s
), 66.7(d).

63.8 (,q), 48.6(d), 48.6
(dL 47.5(d).

47, t(d), 46.8(d,), 43.0
(d), 41.7(d).

38.4(t), 38.0(t), 36.7(
t), 329(d).

28.7(q), 25.6(t), 24.8(
t), 21.5(t).

21.0 (t), 17.4 (C1), 13.0
((1) Mass M" m/z 506 IRv 3380, 1660, 1685 Example 2 Ikarugamay 7 derivative obtained in Example 1 (Compound 1:
(la) In the formula, R= CH3) 101 m9 (
0.2 mmol) and 5 ml of pyridine and dissolve it, 50
It was heated at ℃ for 1 hour. The reaction solution was dried to dryness under reduced pressure, and the residue was recrystallized from acetone to give 90 m9 of colorless needle-shaped crystals of icargamycin visiting conductor (compound 2) represented by the formula (la) where R = -1 (yield: 918%). ) was obtained.

Melting point 240-242℃ UV λEL0H327nm 'H-NMRa ppm (pyridine-d
5) 7.47 (cl, LH), 6.81 (d, d,
LH), 2.81(s, 3H)13C-NMRδ
ppm (pyridine-d5+CD, 0D) 1
95.5(s), 182.9(s), 175.1(s)
, 167.8(s).

142.0(d), 139.4(d), 133
．． 3(d), 131.4(d).

129.4(d), 125.8(d), 103.8(s
), 64.8(d).

49.6(d), 49.5(d), 48.6(
d), 48.0(d).

47.7(d), 43.3(ci), 424(
t), 39.2(d).

39.2(t), 38°0(L), 33.8(
d), 26.7(q).

26.1(t), 26.0(t), 224(t
), 22.1(t).

18.2(q), 13.8(q) Mass M” m/z 492IRyKB”
cm-' 3350.1645.1575 Mitsu IM Shiori 3 Icargamycin 0.969 (2 mmol) dissolved in 60 ml of dimethylformamide and potassium carbonate 1.38 F
l (L OmmoL) 't, in addition, under stirring at room temperature,
Methyl iodide (3.969 (30 mmol)) was added dropwise over about 1 hour, and the mixture was further stirred for 10 hours under the same conditions. Reaction 1' was pressurized to 200 iz of ice water, and the obtained crystals were then recrystallized using an ether-acetone mixture solution in the same manner as in Example 1 to obtain (lb) represented by R1=CH3, R1=-CH3 Colorless crystals of Ikarugamysino derivative (compound 3) (0.389<yield: 37.6%) were obtained.

Melting point 131-133℃ UV λctoo 250 nm (sh,)'H-N
MRa ppm (CDC13) 6.80 (d, d
, LH), 6.30 (d, LH), 2.9
9 (s, 3H).

1.45 (S, 3H) 13C-NMRδ ppm (CDCI!s) 204
．． 6(s), 188.3(s), 169.2(
s), 166.2(s).

154.7(d), 142.5(d), 1312(d)
), 128.0(d).

123.0(d), 121.5(d), 67.0(
d), 65.7(d).

50.7(d), 50.3(d), 48. ．． 1(d),
47.1(d).

46.9(d), 46.7(d), 41.6(d), 3
8.1(t').

38.1(t), 38.1(t), 32.8(d), 2
9.7(t).

27.5(q), 27.5(t), 24. t(
tL 21.4 (t).

t7.4(q), 15.8(q), 13.0(q) Ma
ss M" m/z 5Q6ERv,, x cm 3400.1705.1670.1615Example 4 Ikarugamaishi/derivatives were prepared in the same manner as in Example 3 except that hbenzyl odor was used instead of methyl iodide in Example 13. Colorless crystals of (Compound 4) were obtained.

Melting point 145-146℃ tJV 2°tOR25Qnm (sh,)'H-N
MRδ ppm (CDCe,) 8.27 (br, L
H), 7.13 (s, 5H), 6.80 (d, d, LH
).

6.42 (br, LH), 6.25 (d, IH) 13C
NMRδ ppm (CDCe,) 206.9(s),
187.4 (s), 171.3 (s), 166.9 (s
).

155.7(d), 142.8(d), 134.5(s
), 131°4(d).

130.1(d), 130.1(d), 128.3(d
), 128.3(d).

128.3(d), 127.3(d), 123.5(d
), 122.9(d).

71.5(s), 62.8(d), 50.6(
d), 49.7(d).

48.3(d), 47.4(d), 47.4(
d), 47.0(d).

42.3(d), 38.5(t), 38.1(
t), 38.0(t).

379(t), 332(d), 30.3(t)
, 29°4(t).

25.1(t), 21.7(t), 17.6(
q), 13.2(q).

M ass M” m/z 56
gIRvK11'cm-' 3400.17 to 0,1665.1615 Example 3 l-bromo-
Except for using 3-methyl-2-butene, “# mf+1
1°, :1ril! K L-C(Ib) E-, =-
Hog, = j,, 7- CH2CH==CCH3)2
Compound 5)
Colorless crystals were obtained. -Melting point 128-130℃ EtOI! UV λ 250nm (sh,)'H-
NMRδ ppm (CDCl!s) 8.55 (b
r, IH), 6.78 (d, d, lH), 6.46 (b
r, LH) 6.25 (d, IH), 4.94 (t,
LH), 1.60 (s, 6H) 13C-NMR
δ ppm (CDCe3) 206.9(s), L8
8.1(s), 171.6(s), 166.7(s).

155.1(d), 149.9(d), 136.9(s
), 131.4(d).

128.2(d), 123.4(d), 122.7(d
), 116.2(d).

69.7(s), 62.7(d), 50.9(
d), 50.1(d).

48.4(d), 47.3(d), 47.3(
d), 47.0(d).

42.0(d), 38.4(t), 38.1(
t), 38.1(t).

33.1(d), 30.8(t), 30.2(
t), 30.2(t).

25.8(q), 25.0(t), 21.6(
t), 17.9 (ct).

t7.6(q), t3. t(q) Mass M” m/z 546IRν
3400.1710, 1670.1615 and above Procedural amendment (spontaneous) May 19, 1985 Patent Application No. 81878 2, Title of invention: Icarugamycin Exodus 3, Relationship to the case of the person making the amendment Application Name: Nisnis Pharmaceutical Co., Ltd. 4, Agent name (6870) Patent attorney Miyuki Ariga 1゛□,
, ：Address same as above
Shi-yu.

Name (8632) Patent attorney Nobuo Ono, 1';
, -y6, "Detailed Description of the Invention" column 7 of the specification subject to amendment, Contents of amendment (1) In the specification, page 16, bottom line 2 r (la) In the formula, R = -CHsJ is r(la) in the formula R,=-CI
, correct it as J.

(2) Same, page 18, line 1 r (+,) where R=-CHlJ is replaced by r (1m
) In the formula, correct it as R,=-CH,J.

(3) Same, page 18, line 4 r　In the formula (1m), R=-HJ Correct it to 1(,=-)IJ in the r(1&) formula.

(4) Same, page 19, line 11 r (Ib) where R1=-CH3+ J''-CR3J
A certain r (lb) in the formula R2=CH3+ R3=CH
Correct as 3J〇(5) Same, page 20, line 14 r (Ib) In formula R,=-HI R,=-CH,-C>
J means "(Ib) in the formula R,=-H,R,=-C
H, PeD,” he corrected.

(6) Same, page 21, line 3 from the bottom [(Ib) formula intermediate melting=-HIR,=J is replaced with "(Ib
) in the formula is corrected as R2''-H+ R3=J.

Claims (1)

[Claims] 1. The following general formula (I), ▲There are mathematical formulas, chemical formulas, tables, etc.▼(I) [In the formula, A is a group ▲There are mathematical formulas, chemical formulas, tables, etc.▼ (Here, R_1 is (represents a hydrogen atom or methyl group) or group ▲ Numerical formula, chemical formula, table, etc. An icargamycin derivative represented by: