JPH0372492A - Anthracycline derivative and its production - Google Patents

Abstract

NEW MATERIAL:A compound of formula I (R1 and R2 are each H or lower alkanoyl; R3 is lower alkanoyl). EXAMPLE:2,4,3''-O-triacetyl-2-hydroxyaclacinomycin A. USE:A carcinostatic. PREPARATION:Reaction is made under glycoside-modifying condition between an anthracycline derivative of formula II (R11 and R21 are each H or lower alkanoyl) and a trisaccharide of formula III (O-alpha-L-cinerulosyl-(1 4)-O-(3-O- acyl-2-deoxy-alpha-L-fucosyl)-(1 4)-alpha-L-rhodosamine) followed by, if needed, deacylation.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a novel anthracycline derivative and a method for producing the same. , R1 represents a lower alkanoyl group, and a method for producing the same.

Anthracycline antibiotics have a broad anticancer spectrum against experimental tumors and are widely used clinically as cancer chemotherapy agents. Attempts have been made to create various related compounds by various means such as microbial conversion methods, and some proposals have already been made [for example, Adriamycin (1973)].
-26713 publication or U.S. Patent No. 3.803.124
specification), aclacinomycin A (Japanese Patent Publication No. 54-38
099) 2-Hydroxyaculapinone (JP-A-56-49341) 2-hydroxyaclacinomycin A (JP-A-57-18672), anthracycline derivatives and their total synthesis (P, ^rca
mone, Topics in Ant+biotic
Chemistry, Vol 2. Pages 102 to 279.

εLLIS HORWOOD LIMITε mouth issuance, etc.]
. Among these compounds, adriamycin and aclacinomycin A are widely used clinically as cancer chemotherapeutic agents, but the search for more useful compounds is also underway. Some of the present inventors also used 2-hydroxyaclacinomycin A represented by the following formula as a kind of useful compound using a microorganism of the genus Streptomyces to obtain aglycone (2-hydroxyaclavinone) B, and then Furthermore, it is produced by glycosylation using another type of microorganism belonging to the genus Streptomyces, or it is produced by direct fermentation using a microorganism obtained by cell fusion of a microorganism capable of producing the above aglycone and a microorganism capable of glycosylating the aglycon. The present invention was also completed for the purpose of providing a semi-synthetic method for 2-hydroxyaclacinomycin A and its derivatives (see JP-A-58-26895). The anthracycline derivative represented by the general formula (1) provided by the invention, wherein R1 and R2 each independently represent a hydrogen atom or a lower alkanoyl group, and R3 represents a lower alkanoyl group, is an anthracycline derivative represented by the above-mentioned 2-hydroxy Not only can it be used as a synthetic precursor to adalacinomycin, but it also has excellent antitumor activity as described below, and can be used as an anticancer agent.

As used herein, the term "lower" means that the group to which this term is attached has no more than 6 carbon atoms, preferably no more than 4 carbon atoms.

Therefore, in the -monhole (1), R+R2 and R
The "lower alkanoyl group" represented by 3 is an alkanoic acid residue in which the alkyl moiety is linear or branched and has 1 to 5 carbon atoms, preferably 1 to 3 carbon atoms, such as acetyl, propionyl, etc. , n-butyryl, imbutyryl.

Included are n-penthiol, isopenthiol, and the like.

An acetyl group is particularly suitable as such a lower alkanoyl group.

The compound of formula (1) provided by the present invention is obtained by adding 2-hydroxyaclavinone prepared by the above-mentioned fermentation method (see JP-A-56-49341) or total synthesis method in the presence of boric acid. By reacting with a lower alkyl carboxylic acid anhydride, among the hydroxyl groups active in the glycosylation reaction described below, the 2- and 4-position hydroxyl groups, which are inconvenient for the reaction, are selectively removed.
-By acylation, an aglycone derivative represented by the formula (II) in which R+ and Rz have the same meanings as above and the general formula (III)H in which R1 represents a lower alkanoyl group , trisaccharides (〇-α-L-sinellosyl(1→4)-0-(
3-0-acyl-2-deoxy-α-L-fucosyl)-
(l→4>-α-L-rhodosamine) under glycosidation conditions to produce the anthracycline derivative represented by the above-mentioned (1), and this compound is further subjected to a partial deacylation reaction. By doing so, each can be obtained.

The unit reactions at each stage shown in the above reaction are known per se, and can be carried out according to known methods. −
The glycoside reaction with 2,4-0-diacyl-2-hydroxyaclavinone shown by Monka (II) first begins with -
The trisaccharide (III) (1 to 5 equivalents, preferably 2 to 3 equivalents based on the aglycone derivative) is dissolved in an inert solvent such as dichloromethane chloroform or tetrahydrofuran at 0°C to -70°C (preferably -30℃~-
(at 70°C) pyridine compounds with alkyl side chains such as 2,4,6-collidine and 2,6-lutidine, or bases such as N,N-dimethyl-〇-)luidine (1 to 15
, preferably 4 to 1 g equivalent), in the presence of a sulfonylating agent, such as an acid anhydride or acid chloride of trifluoromethanesulfonic acid, methanesulfonic acid or p-)luenesulfonic acid (preferably 1 to 1. 5 equivalents> to convert the 1-position hydroxyl group of the sugar into the desired sulfonate, and then the quaternary ammonium salt of bromine (3 to IO
1 (preferably 4 to 6 equivalents), for example tetra-n-butylammonium bromide, to convert it into bromo sugar. Thereafter, a suitable solution of 2-hydroxyaclavinone, such as a dichloromethane solution, is added to carry out glycosidation. After gradually increasing the reaction temperature and stirring at 0°C to room temperature until the consumption of raw materials stops, the reaction solution was mixed with chloroform, dichloromethane,
Ethyl acetate.

Dilute with a solvent such as benzene, wash well with 5% K H, P O, aqueous solution and water, and remove the organic layer with anhydrous NazS04.
After drying using a desiccant such as, etc., the residue is obtained by drying under reduced pressure. When this is separated and purified by chromatography, for example, a silica gel column or thin layer chromatography using a chloroform/methanol-based developing solvent, the desired 2,4.3''-〇-)lyacyl- 2
-Hydroxyaclacinomycin is obtained.

In addition, the glycosidation reaction at this stage is carried out in the above-mentioned -Momona (II)
It can also be carried out by reacting a reactive derivative of the trisaccharide of I) under suitable glycosidation conditions. In this case, the reactive derivatives include, for example, a glycal product in which the 1-position hydroxyl group and 2-position hydrogen atom derived from rhodosamine of the trisaccharide represented by formula (I[I) are dehydrated, and the 1-position hydroxyl group derived from rhodosamine. Examples of glycosidation conditions include acidic conditions,
Glycosidation conditions known per se, in which the compound of the above-mentioned -Momonena (I[) and the reactive derivative are reacted in the presence of a Lewis acid such as tin tetrachloride or an organic sulfonic acid such as l)-toluenesulfonic acid ( For example, Carbohy
draLe Re5earch, 101. Chapter C1
~C4, 1982).

The above glycosidated raw acids can be deacetylated. For example, 2,4.3″-〇-triacetyl-2-hydroxyaclacinomycin 8 is dissolved in a solvent such as tetrahydrofuran, and 0.1N-Na is dissolved at -10°C to 5°C.
Add 3 to 5 equivalents of y'9/- solution of OH, and add 10 to 60
Stir for a minute. After that, acetic acid, KH, PO,, HCl
The crude product is neutralized with an aqueous solution of chloroform, benzene, ethyl acetate, etc., and the extract is washed with water and dried under reduced pressure to obtain a crude product.

When this is purified by chromatography, partially deacylated 3''-〇-acetyl-2-hydroxyaclacinomycin 8 can be obtained.

Furthermore, when the above methanolysis is carried out at room temperature for 1.5 to 3 hours, completely deacetylated 2-hydroxyaclacinomycin can be obtained. The physicochemical properties and biological activities of the purified product of this substance were consistent with those of 2-hydroxyaclacinomycin obtained by fermentation.

Note that the trisaccharide used in the reaction in this step is O-α-L-sinellosyl(l→4)-0-(2-deoxy-α-L
-fucosyl)-(l→4)-α-L-rhodosamine, such as the aforementioned aclacinomycin Δ, sinerpine Δ (＾nstimic
robial Agents and (he+noth
erapy-1970, pp. 68-77, 1
It can be easily prepared by acylating the hydroxyl group at the 3'' position of 971) etc. by a known method, and then purifying and collecting the a part released from catalytic reduction using palladium-barium sulfate as a catalyst.

In addition, in the compound of -Momonena (f), the compound (3''-〇-acyl-2-hydroxyaclacinomycin A) in which R1 and R2 represent a hydrogen atom can be directly converted to 2-hydroxyaclavinone by the formula (III). It can also be obtained by reacting trisaccharides.

The compound of general formula (1) obtained as above has excellent antitumor activity as described above, and exhibits almost the same activity as 2-hydroxyaclacinomycin obtained by fermentation method. .

The antitumor activity of the compound (1) provided by the present invention can be demonstrated by the experiments described below.

The compound of the present invention can be used in mouse leukemia cultured cells (L1210
) and markedly inhibits the proliferation and nucleic acid synthesis. for example,
Incubate L1210 cells 5X in RPMI 1640 medium (Rothwell Park Laboratories 1640) containing 20% calf serum.
I (1'/rnl) was inoculated, and at the same time the compound of the present invention was added at a concentration of 0.01 to 0.5 μg/-, 37
The cells were cultured in a carbon dioxide gas incubator at ℃ for 48 hours, and the 50% growth inhibition concentration relative to the control group was determined. Furthermore, the above L121
0 cultured cells were suspended in RPMI1640 medium containing 10% calf serum to a concentration of 5 XIO' and incubated at 37°C.
After culturing in a carbon dioxide gas incubator for 1 to 2 hours, the compound of the present invention was added at various concentrations, and after 15 minutes, an additional 1 hour incubation was carried out.
C-uridine (0,05 μCi/−) and 4C-thymidine (0,05 μCi/mf) were added and incubated at 37°C for 60
Incubate for minutes. 10% trichloroacetic acid solution was added to the reaction solution to stop the reaction, and at the same time, to precipitate acid insoluble materials,
After further washing three times with 10-5% trichloroacetic acid, it was dissolved in formic acid, the radioactivity in the acid-insoluble matter was measured, and the 50% uptake inhibition concentration was determined from the radioactivity uptake rate with respect to the control group. The results are shown in Table 1.

As is clear from the experimental results described above, the compound (I) provided by the present invention is L1210
Superior proliferation and DNA and RNA against leukemia cells
It exhibits a synthetic action and can be predicted to have excellent antitumor activity.

The present invention will be explained in more detail with reference to production examples below. Incidentally, since 2,4-0-diacyl-2-hydroxyacrabinone is a raw material for the production method of the present invention and is a new compound, an example of its production will be shown.

Production example 1: 2.4-0-Diacetyl-2-hydroxyaclavinone 120 mg (0.28 mmol) of 2-hydroxyaclavinone and 0.51 mg of acetic anhydride containing B(○H) = 52 mg (0.84 mmol) (5.04 mmol) and dissolved by adding 2,4-tetrahydrofuran. After stirring at 23°C for 30 minutes, pyridine 0.0
27mj! (0.34 mmol) was added to complete the reaction (23°C, 30 minutes).

Next, the reaction solution was poured into 20 d of ice water, and 30 ml of chloroform was added.
The chloroform extract was extracted with water (30mj!x
2>, washed with 2% Na HCOs aqueous solution (30 mg) and again with water (30 mg), dried over anhydrous NatSOa, and evaporated to dryness. The resulting yellow residue was crystallized using silica gel column chromatography (7 g of silica gel, chloroform) from chloroform/n-hexane to give 125.4 mg of 2.4-0-diacetyl-2-hydroxyaclavinone (87, 3%).

mp, : 113-115°C [α) l:' : +90.7° (c = 0.03
．． CHC1,) UV-Vis (λ, nm
, (E”%))1CflCls maw 1cm260(5
04,7), 286 (213J), 417 (169,3
), 432 (170,0) IR (KBr, am-'): 1780, 1735. 1680. 1625NMR
(CDCis, δppm): 1.08: C11
3 (t, J=6Hz), 1.5 ~ 2.35:CH
*X2 (m).

2.35:C0C)Is(S>, 2.45:C0C
L(s), 3.48:'l-roH(d, J=3.5
H2), 3.69:C00CH3(S), 3.
88:9-RoH(S), 4.05'1(IH(S)
, 5.32:7-H (bs, 11t+ =811z)
.

7.23:3-H (d, J=2.5Hz), 7.61:
If-H(s), 7.96:1-H(d, J=2.
5Hz), 13.25:6-0H(s) Example 1:
2.4,3”-0-)lyacetyl-2hydroxyaclacinomycin A O-α-L-cinellosyl-(1→4)-0-(3-0
-acetyl-2-deoxy-α-L-fucosyl)-(l
→4) -α,β-ri 87.5mg of dosamine (0,
19 mmol>, tetra-n-butylammonium bromide 122B (0.38 mmol) and 2° 4,6-collidine 0.075 mj! (0.57 mmol) in dichloromethane was cooled to -60°C. , trifluoromethanesulfonic anhydride 0.042 rnl (0°24
After 10 minutes, 65 mg of 2.4-0-diacetyl-2-hydroxyacrapinone (0,127
A dichloromethane solution (1rn1) of 1 mmol) was added using a syringe, the reaction temperature was gradually raised, and the mixture was stirred at room temperature for 1 hour. The reaction solution was again cooled to -60° C., and the same amount of freshly prepared glycosidation solution as above was poured into the solution, followed by reaction at room temperature for 2 hours. Next, the reaction solution was mixed with 30 In of benzene! Dilute with
Water (30dX2), 5% KHz PO4 aqueous solution (30mt
'X2), after washing with water (30dX2) again, anhydrous Nat
It was dried with SOn and evaporated to dryness. The obtained residue was purified by silica gel column chromatography (5 g of silica gel). First, develop with chloroform and unreacted 2,
410-Diacetyl-2-hydroxyactabinone 29
．． 7*g (4.57%) was recovered and the title compound 2,4.
3″-0-)lyacetyl-2-hydroxyaclacinomycin:/A 36.2mg (29.9%), further o
5.7 mg (4.9%) of the 2-position deacetylated compound 4.3''-〇-diacetyl-2-hydroxyaclacinomycin A was obtained from o-form/methanol (20/l).

2.4.3″-o-triacetyl-2-hydroxyaclacinomycin A: mp, : 1'37~140°C (α1g': 113.3° (c~0.0
．． 3. CHCA, ) HCl3 UV-Vis (λ, nm, (E1%))
:max 1cm 259 (267,7), 266 (268,3), 418
(82,0).

438 (74,0) IR (KBr, cm-'): 1780, 1730. 1675. 163ONMR
(CDCis, δppm): 1.0 ~ IJ5
:CLX4(m), 2.08:COCl1s(s)
.

2.17:N(CL)z(s), 2,37:COC
l1*(s), 2.49:C0C)+3 (S
), 3.66:C00CH, (s), 4.
(19:1O-)1(s).

4.95-5.55:7-H,I'-)1. 1″-H
, 1'''-N, 3''-H(a+), 7.39:3-
H(d), 7.64:1l-H(s), 7.98:1-
)1(d), 13.24:6-014(s)4.3
"-0-Diacetyl-2-hydroxyaclacinomycin A mp,: 167-171°C Cal g': -110,5° (c ~ 0.04
．． CHCl 5) LIV-Vis(λ,
nm, (E'%)): CllCl.

max1cm 270 (320,8), 41B (94,0), 44
0(82,4)I R(KB r, cs-'):
1770, 1730. 1675.1625NMR(C
DCis, δppm): 1.0 to 1.4: CHs
X4(m), 2.09:C0CL(s).

2.29:N(CHs)*(s), 2.38:C0C
H5(S), 3.75:C00C113(S), 4
, 10:1O-H(s), 4.95-5.55:7-
H.

1'-H, 1"-H, 1°"-H, 3"-H (a),
6.58:3-H(d).

7.17:1-H(d), ? , 41:1l-H(S)
, 13.30:6-0H(s) Example 2:3″-O-
Acetyl-2-hydroxyaclacinomycin A 2.4.3'-0-)Lyacetyl-2-hydroxyaclacinomycin p, 20 mg (0,021 mmol) in dry tetrahydrone 2InI! Dissolved at 0℃
Cool to 0. A 0.84-mL methanol solution of IN-N a 0CHs was added. After 20 minutes, the reaction solution was reduced to 5% K.
H.

The mixture was neutralized with an aqueous solution of PO, diluted with 20% of chloroform, washed with water, dried over anhydrous Na 2 SO, and evaporated to dryness under reduced pressure. The obtained residue was subjected to silica gel column chromatography (4 g of silica gel, chloroform/methanol (
20/l)) and purified with 3" O-acetyl-2-hydroxyaclacinomycin A 13.4+ng (73
,6% n was obtained.

mp,: 169-170°C (XE +t': 42.5°
(c ~0.04.CHC1,n('1lcI
3 UV-Vis (λ, nm, (E'%))
:max 1cm 270(240,0), 2B? (210,4), 4
42(124,4)[R(KBr, cl'): 1735. 1675. 1625 NMR (CDCf, δppm): 1.0 to 1.
3: CHs x4 (m), 2.08: C0CH5 (S
)2.22:N(CHs)2(s), 3,73:C00
CH,(s), 4.10:10~H(s), 4
．． 9-5.6:7-H,1°-H21″-H,l°″-
H23”-H(m), 6.26:3-H(d), 6
．． 74:1-1 ((d), 7.41:11)1(S),
11.92: A r OH (br), 12
．． 64: ArOH (br) Example 3: 2-hydroxyaclacinomycin A2.4
．． 3″-〇-) lyacetyl-2-hydroxyaclacinomycin Δ1 gmg (0,019 mmol) was dissolved in 1.8-ml of dry tetrahydrofuran at 0°C under nitrogen.
0. Methanol solution of IN Na OCHs 0.
After adding 76-, the mixture was reacted at 15 to 20°C for 2 hours. The reaction solution was neutralized with a 5% K 82 P 2 O aqueous solution, washed with water by adding 20 ml of chloroform, dried over anhydrous NaxSOI, and then dried under reduced pressure to obtain a yellowish brown powder.

This was subjected to preparative thin layer chromatography (silica gel 60
．． F-254, Merck & Co., Ltd., chloroform/methanol (7/1)) to give 10.2 mg (65.4%) of 2-hydroxyaclacinomycin A.

mp,: 166-167℃ [α]g': +43.2' (cm0.04.M
eOH) UV-Vis (λ, nm, (E
'%)).

IICI3 max 1cm 222 (369), 256 (231), 295
(204), 450(108)IR(KBr,
cm-'): 1735, 1675. 1620. 1611)NM
R (CDCis, δppm): 1.0 to 1.
4: CL x 4 (m), 2.30: N (CL) 2 (s
).

3.72:C mouth OCI+3(S), 4.08:1O
-OH(S), 4.95 ~ 5.60:7-H, 1
°-N, 1'-H,, 1''-)1(m), 6.
24:3-H(d), 6.73:1-H(d), 7.3
0:1l-H(s) Literature value (The Journal
of Antibiotics, 34°916-91
11. 1981) mp, : 165~167℃ (ff]H”: +42.3° (cm0.04.M
eOH) UV-Vis (A90%IJ eOH
, nm > max 222, 256. 295. 450IR (KBr,
cm-'): 1735, 1675. 1620. 1610ffJ
1! i Example 4: 3″-〇-acetyl-2-hydroxyaclacinomycin A 〇-α-L-cinellosyl(l→4)-0-(3-O
-acetyl-2-deoxy-α-L-fucosyl)-(1
-4) -α,β-L-Rhodosamine 138mg (0,3
mmol) 'l, 4.6-collidine 0.118
A solution of ml' (0.9 mmol) in 3 ml of dichloromethane was cooled to -50° C. and 0.05+nf (0.3 mmol) of trifluoromethanesulfonic anhydride was added.

After stirring for 5 minutes, the solution was mixed with 42.7 mg (0.1 mmol) of 2-hydroxyaclavinone, 193 mg (0.6 mmol) of tetra-n-butylammonium bromide and 1.0 g of pimply sieves 4A in 10 rn of dichloromethane. ! solution (cooled to -50°C). After raising the reaction temperature and stirring at 20°C for 1.5 hours, Example 1
A crude product was obtained by post-treatment in the same manner as above, and this was purified by silica gel column chromatography (5 g of silica gel, 1 chloroform/methanol (40/1)) to obtain L, 3''-〇-acetyl-2-hydroxyaclacinomycin from A. A
6 mg was obtained (which showed the same physicochemical properties as the product of Example 2).

Claims (1)

[Claims] 1. General formula ▲ There are mathematical formulas, chemical formulas, tables, etc. ▼ In the formula, R_1 and R_2 each independently represent a hydrogen atom or a lower alkanoyl group, and R_3 represents a lower alkanoyl group. Anthracycline derivatives. 2. General formula ▲ There are mathematical formulas, chemical formulas, tables, etc. ▼ In the formula, R_1_1 and R_2_1 represent a hydrogen atom or a lower alkanoyl group.
There are chemical formulas, tables, etc. ▼ In the formula, R_3 represents a lower alkanoyl group. A general product characterized by reacting a sugar residue shown by or a reactive derivative thereof under glycosidation conditions and deacylating it if necessary. A method for producing an anthracycline derivative represented by the formula ▲ There are mathematical formulas, chemical formulas, tables, etc. ▼ In the formula, R_1 and R_2 each independently represent a hydrogen atom or a lower alkanoyl group, and R_3 represents a lower alkanoyl group.