DE3241199A1 - (ALPHA), (ALPHA) -TREHALOSE-6,6'-DIESTER MEDIUM CHAIN ALIPHATIC ACID, AND PHARMACEUTICAL AGENTS WITH A CONTENT THEREOF - Google Patents

Description

Z.

1A-4070

FP-SS-3

SS PHARiIACEUTICAL CO., LTD. Tokyo, Japan

cc, ct-trehalose-6,6'-diester medium chain, aliphatic Acids as well as pharmaceutical agents with a content

the same

The present invention relates to new α, α-trehalose-6,6'-diesters medium chain aliphatic acids as well as pharmaceutical agents which show them to be effective Part included.

α, α-Trehalose is one of the so-called disaccharides ^ which consist of two glucose molecules are composed and are widespread in the field. Various derivatives

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of these compounds have been synthesized and examined for their pharmacological activities. So For example, it has been reported that a mixture of different diesters in which the aliphatic acids are attached to different Positions are bound to show antitumor activity.

In the course of the synthesis of various β, β'-aliphatic Acid diesters of α, α-trehalose and in the investigations Its pharmacological effects have been found by the inventors to be a cc, cc-trehalose-6,6'-diester medium-chain, aliphatic acids according to the following formula (I)

CH ₃ (CH ₂ J _n COO

where η is an integer from 6 to 10, excellent antitumor activity, excellent antibiotic activity and an excellent effectiveness in terms of delay-type hypersensitivity and is useful for pharmaceuticals. The present invention is based on these studies.

It is accordingly the object of the invention to provide a new cc, cc-trehalose-6,6'-diester medium-chain, aliphatic acids according to the above formula (I) are available place. It is also an object of the invention to provide a pharmaceutical agent which is effective Part of an oCjOc-Trehalose-G, 6'-diester medium-chain, contains aliphatic acids according to the above formula (I).

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The inventive a, a-trehalose-6,6'-diester medium-chain, of aliphatic acids (I) (hereinafter referred to simply as "trehalose diesters of medium-chain, aliphatic Acids ") can be prepared, for example, according to the following reaction scheme. Thereafter, α, α-trehalose (II) in 2,3,2 '^' - tetra-o-benzyl-ocjOc-trehalose (III) transferred and then with a medium-chain, aliphatic acid (IV) (the term "medium chain" means here and below an aliphatic acid Remainder of medium chain length) or its active derivative to a new intermediate 2,3,2 ', 3'-tetra-above-zyl-a, a-trehalose-6,6'-diester medium-chain, aliphatic acids (V) implemented and this will continue using a reducing catalyst, reducingly debenzylated:

OBn

OBn

CH ₃ (CH ₂ ) _n COO-n

CH _JL (CH ₂ ) _n COOH fc _B \

(IV) ⁵ ^ HO ¹ -! * ⁰

OBn

Reduction catalyst

(D

where Bn is a benzyl group and η is the one given above Has meaning.

In the above procedure, the 2,3,2 ', 3'-tetra-above-zyl-a, cc-trehalose according to the formula (III) prepared by the trehalose according to the formula (II) with benzaldehyde in a conventional manner to form 4,6, 4 ', 6'-di-o-benzylidene-a, α-trehalose and thereafter this compound with benzyl chloride to form a 2,3,2 ', 3'-tetra-o-benzy! derivative reacts and finally reacts the same with an acid.

The reaction between the compound (III) and the medium-chain, aliphatic acid (IV) or its active derivative under conventional conditions undergoes an esterification reaction performed by using the medium-chain, aliphatic acid (IV) or its active derivative in a molar ratio of preferably 1.8 to 2.4 mol / mol the compound (III) in the presence of an acid binder, such as pyridine and triethylamine, is reacted.

The reaction is preferably carried out in the presence or absence of a solvent between ⁰ ° C. with ice cooling to 30 ^° C. for 1 to 24 hours. The solvent used therein includes, for example, methylene chloride, chloroform, benzene, toluene, ether, tetrahydrofuran and dioxane.

The reducing debenzylation reaction to which the thus obtained compound (V) is subjected is carried out by means of a catalyst, preferably in an amount of 0 _? 5 to 10 mol / 1 mol of compound (V) in a suitable solvent.

The reducing catalyst includes, for example, palladium in ear, palladium-carbon and Raney nickel. The solvents used include, for example, alcohols, such as methanol, ethanol and isopropanol; halogenated compounds such as chloroform and methylene chloride; as well as mixtures this solvent.

The pharmacological activities of the invention Trehalose diesters of medium-chain, aliphatic acids (I) are shown below.

(1) Cell-killing activity of trehalose diester of medium-chain aliphatic acids

(a) Leukemia L-5178Y cells are used and adjusted in such a way that 1Cr cells / ml are present in an RPMI 1640 medium containing 10% ox fetal serum. The samples are applied to the medium up to a predetermined concentration. After cultivation in an incubator at 37 ° C. under 5% COp for 4-3 h, the number of living cells is counted under a microscope and compared with a control group in order to determine the multiplication factors. These are plotted on a logarithmic probable paper and the values for ICj-Q v / earth are determined based on the curve obtained in this way. The concentration for each sample is 25, 50, 100, 200 and 'respectively. 400 / µg / ml

The results are shown in Table 1.

Table 1

Test connection [an- IC, - ₀

give is the number for °

η in formula (I)] (yug / ml)

6,350 according to the invention

Connections 3 255

10.93

Comparison compounds 14> 400

20> 400

(b) In the same way as under (a), ground the cell killing activities the trehalose diester of medium-chain, aliphatic acids against cells of Leukemia L-1210 examined. The results are shown in Table 2.

Table 2

Test compound [an- - ^ ro

give is the number for

η in formula (I)] (/ Ug / ml)

6,350 according to the invention

Connections 8 150

10 210

Comparison compounds 14> 1000

20 _> 1000

(2) Anti-cancer activities of the trehalose diesters of medium-chain, aliphatic acids

Male std-ddy mice, 5 weeks old, are used namely six mice / group each. Sarcoma 180 cancer cells are found in the peritoneal cavity of each mouse transplanted, namely 5 × 10 cells. Starting 24 h later, a test compound is suspended in 30? ά Propylene glycol, administered intraperitoneally in an amount of 150 mg / kg once a day continuously for 5 days. Samples of the abdominal ascites are taken 7 days after the cancer cells are transplanted to reveal the Amount of abdominal ascites and the cell ratio

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* m Jr \

to determine. The tumor growth ratio (T / C in %) is determined using TPCV (total pack cell volume), which is the product of the amount of abdominal ascites and the cell ratio according to the following formula. The anticancer activity of the test compound is assessed. The results are shown in Table 3.

} TPCV for the treated group ^{; =} TPCV for control group ^x

Table 3

Test compound [indicated tumor growth rate is the number for η ratio T / C in formula (I) 1 (%)

6 according to the invention 9.1

Connections 8 1.9

10 6.2

Comparative compounds 14 37.7 20 55.8

Control group 100

^{T. <T} ie from the above description, v / eist the trehalose diesters of medium-chain aliphatic acids, a remarkably stronger anti-cancer activity than the corresponding compounds having 6 to 22 carbon atoms.

(3) Antimicrobial Activity

The minimum inhibitory concentrations (MIC) of the compounds according to the invention are investigated against various bacteria. The results are shown in Table 4. The tests were carried out according to the IIIC measurement method approved by the Japan Society of Chemotherapy. The Mueller-Hinton medium (Difco Laboratories, USA) was used as the culture medium. The test compounds were dissolved in 25% DMSO to concentrations of 1 mg / ml and then treated with sterile, distilled

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Hard water diluted to the predetermined concentrations .

Results: Table 4

MIC values 12, (/ Ug / ml) comparison
14 20 7 100 Test compound [spec
give is the number of η
in formula (I) 1 invention
6 8 12, > 100 Bacterial strains 7100 5 MOO 7 100 Bacillus subtilis
ATCC 6633 50 7 100 5> 1OO 7 100 Staphylococcus aureus
ATCC 6538P 50 > 100 > 100 > 100 Staphylococcus et> idermi-
of ATCC 12228 100 7 100 Streptococcus faecalis var
ginogenes IFO 3939 * 50 7 100 Sarcina lutea ATCC 9341 100

(4) Effect on delay type hypersensitivity (delayed-type hypersensitivity)

Several groups of 6 week old female ICR mice, Each group of ten mice are provided and given red blood cells subcutaneously of the sheep (SRBC), 0.05 ml / mouse at a concentration of 3 x 10 cells / ml. The injection takes place in the sole of the foot of the right hind paw, with the test compounds immediately thereafter intraperitoneally injected (day 0). 4 days later, the same amount of SRBC (0.05 ml / mouse) of each mouse is in the sole of the foot The left hind paw is injected subcutaneously, and a further 24 h later the thickness of the left sole of the foot is measured using a Measuring circle measured.

Other groups that instead of the test compounds are physiological Saline is used as controls, and other groups only subjected to the second injection

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used as a reference (base). The swelling ratio of the sole of the foot is obtained from the following equation:

Swelling ratio (9ό) = £ ~ "T ~ § ^{x 10} ° where T, C and 3 are each the thickness of the sole of the left hind paw of the test groups, the control groups" and of the reference groups mean. The results are shown in Table 5.

Table 5

Test compound; an- thickness
given is the number
of η in formula (I) the sole of the foot
(R.A.M) Swelling
ratio (5o) Compounds according to the invention 6th
3
10 3.71
3.75
3.82 . 126
133
138 Comparison compounds 14th
20th 3.61
3.55 109
98 control 3.56 100
.It

Dose: 100 mg / kg; Vehicle: normal saline + 1 drop of Tween 80

The trehalose diester according to the invention of medium-chain, aliphatic acids is almost non-toxic and so represents a safe connection, which is also from the fact It is clear that, for example, sucrose esters of aliphatic acids which are similar to the esters according to the invention are approved as food additives and are widely used Scope as harmless surfants in the field of food, drugs, cosmetics or the like. be used.

The compounds according to the invention can also lead to various Types of preparations are formulated, depending on the type of application, such as oral or non-oral administration. For example, oral

ORIGINAL BATHROOM

- Λ Λ _%

administrable preparations such as tablets, capsules, Powders, granules and liquids, as well as formulations that cannot be administered orally, such as injection solutions for subcutaneous, intramuscular or intravenous injection, transfusions and suppositories and the like will.

The above-mentioned preparations can be produced according to any known method. This means, The trehalose diester of medium-chain, aliphatic acids can be in tablets, capsules, powders or granules be deformed by the compound optionally in combination with excipients such as starch, lactose and Mannitol; or binders such as sodium carboxymethyl cellulose and hydroxypropyl cellulose; Disintegrants such as crystalline cellulose and calcium carboxymethyl cellulose; Lubricants such as talc and magnesium stearate; Fluidity improve, such as light, anhydrous silica and the like., formulated. Liquids and injections can also be prepared by treating the trehalose diester medium-chain, aliphatic acids in vegetable oils or the like. To form an oil-soluble injection solution or by conventionally dissolving the compound in water or the like to form a Dissolves or suspends syrups. In addition, suppositories can be made by making the compound in a commonly used substrate, e.g. cocoa butter, synthetic oils and fats or the like, dispersed and then solidified.

Including the amount in the application of the invention Trehalose diester of medium-chain, aliphatic acids depends on the degree of the disease, it will preferred, in adults the compound at a dose of 0.1 to 2000 mg / kg for oral administration and 0.05

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- Ψ- ix.

up to 1000 mg / kg for non-oral administration, either all at once or several times in portions during the day.

The invention is explained in more detail below with the aid of examples.

example 1

20 ml of pyridine and 20 ml of dichloromethane are added to 2.11 g of 2,3,2 ', 3'-tetra-o-benzyl-α, a-trehalose and the whole is dissolved. While cooling with ice, 1.26 g of decanoyl chloride are added dropwise ^{at 0 ° C. with stirring.} After stirring for 30 minutes, the temperature is raised to room temperature and stirring is continued for an additional hour. The reaction mixture obtained in this way is poured into ice-water and extracted with chloroform. This is then distilled off under reduced pressure. The residue is purified by silica gel column chromatography. 2.79 g of 2,3,2 ', 3'-tetra-o-benzyl-6,6'-di-o-decanoyl-a, a-trehalose are obtained as a colorless, oily product (yield 3Z%) .

Example 2

2.67 g 2,3,2 '^' - Tetra-o-benzyl-ö ^ '- di-o-decanoyl-ajtttrehalcse, obtained in Example 1, v / earth dissolved in 40 ml of chloroform, and the solution is catalytic for 1 h Subject to reduction. To this, 1.00 g of palladium black are added as a catalyst and hydrogen gas initiated into the solution. The reaction solution obtained in this way is filtered to separate off the catalyst and the filtrates are concentrated and concentrated to dryness under reduced pressure. The residue is recrystallized from isopropanol. 1.06 g of 6,6'-di-o-decanoyl-oc, cc-trehalose are obtained as colorless, needle-shaped crystals (62% yield).

Example 3

The compounds listed in Table 6 below are obtained in the same manner as in Example 1. Table 6 also contains the compound obtained in Example 1.

Example 4

The compounds listed in Table 7 below are obtained in the same way as in Example 2. The Table 7 also contains the compounds obtained in Example 2.

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Example 5 Tablets

A tablet is produced in the conventional way, which contains the following ingredients in the specified amounts.

mg

Trehalose diester medium-chain aliphatic acid (decanoate) 100

D-mannitol - 150

crystalline cellulose 50

Strength 23

Calcium Carboymethyl Cellulose 16

Talc 4

Magnesium stearate 2

Total amount 350 mg

Example 6

Capsules

A granulate is produced in the conventional way, which contains the following ingredients in the specified amounts. The granules are listed in No. Capsules filled.

Trehalose diester medium-chain, ali-

phatic acid (caprylate) 25

crystalline cellulose 17

light, anhydrous silica 7

Magnesium stearate '1

Example 7 In.ielctions solutions

A solution for injection is made by the following ingredients each formulated in the specified denominations in a conventional manner.

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Trehalose diester medium-chain, ali-

Phatic Acid (Decanoate) 25 mg

made up to 1 ml with olive oil

Example 8 Suppositories

The following ingredients are each melted and stirred in the specified amounts and then shaped to solidify into a piece of suppository using conventional technique will,

Trehalose diester medium-chain, ali-

phatic acid (laurate) 100

Cocoa butter 1100

Total amount 1200 mg.

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Claims (2)

3241139 Claims a, cx-Trehalose-6,6'-diester medium chain, aliphatic Acids according to the following formula (I) CH ₃ (CH ₂ ) _n COO where η is an integer from β to 10. 2. Pharmaceutical agent, characterized in that it is an α, α-trehalose-6,6'-diester as the active ingredient medium-chain, aliphatic acids according to the following formula (I) CH ₃ (CH ₂ ) ^ OH contains, where η is an integer from 6 to 10.