JPS60115574A - 1,2-dithiolane, manufacture, medicine and use - Google Patents

Abstract

(57) [Summary] This bulletin contains application data before electronic filing, so abstract data is not recorded.

Description

Detailed Description of the Invention [Industrial Field of Application] The present invention provides 1,2-dithiolane-
This invention relates to 1-oxide-carboxylic acids, their gostyls, amides and salts thereof.

The present invention is particularly characterized by the formula (■): ↓ (wherein R1 is hydroxy, lower alkoxy, amino,
represents cycloalkylamino/or 7- or di-lower alkylamino, where each alkyl moiety may be substituted or unsubstituted by aryl, heteroaryl or cycloalkyl, and n is from 0 to 1o.
is an integer of ) in free or salt form.

[Conventional technology]

Various free acids and alkyl esters and salts of the above compounds - e.g. Tetrahedron Letter
s13, 1073-5 (1973); Arch
, Biochem.

Br0P11YS, 185.576-83 (1978
); J, Org.

Chem, 40.58-62 (1,975); Ark
iv, Kemi25 (14-4), 263-77 (19
66); 6,8-dithiok△tanic acid amide monoxide, which is also related to JP-A No. 11.7616/1973, is disclosed in JP-A No. 39-1.0931
listed in the number. There is no particular description regarding the pharmaceutical uses of these compounds.

Surprisingly, it has now been found that (1), 2-dithiolane oxide carboxylic acid and its derivatives have pharmacological activity, particularly immunostimulatory activity as described below. Therefore, in this invention, especially 1,2-dithiolane-1
-Pharmaceutical compositions of oxidecarboxylic acids-or their pharmaceutically acceptable esters-amides or salts-and their compounds for use as pharmaceuticals, in particular as immunostimulants.

Among other things - the invention relates to pharmaceutical compositions comprising compounds of formula (I) as described above - or, where appropriate, pharmaceutically acceptable salts - and to compounds thereof for use as medicaments, in particular as immunostimulants. Pertains to.

Formula (IP): ↓ (wherein -R'1 represents cycloalkylamino or mono- or di-lower alkylamino, in which each alkyl moiety is substituted by aryl and heteroaryl is substituted by cycloalkyl - or unsubstituted) (or n is an integer from 0 to 10) are novel and are likewise included in the present invention.

The compounds of formula (■,) also consist of a preferred group of compounds of formula (I).

The compound of formula (Ip) can be obtained by oxidizing the corresponding compound of the formula (Ilp): (wherein and n have the meanings given above).

According to the invention, the process can use conventional oxidation reactions - for example using m-chloroperbenzoic acid.

The reaction may be carried out - preferably in an inert solvent such as a chlorinated hydrocarbon such as dichloromethane - preferably at a low temperature, e.g. water-cooled. The desired product can be isolated and purified by conventional methods.

Other compounds can be prepared analogously - or according to the methods described in the above-mentioned literature.

The compound of formula (I) generates a syn type and an anti type, giving rise to optical isomers. The mixture obtained by the reaction can be separated by conventional methods such as chromatography.

If a compound exhibits a particular isomeric form, this should be indicated in each case.

Some of the compounds of formulas (■, or alkyl, which may be substituted or unsubstituted by cycloalkyl-heteroaryl or aryl, and furthermore either one of 2 and R3 may represent hydrogen).

These two reaction operations are carried out by conventional methods as in the Examples described below. The compound of formula (1) is also novel and constitutes one aspect of the present invention. Other intermediate products are known or produced in a manner analogous to known procedures.

The intermediate product may be separated and purified by a conventional method, or may be used directly in the next reaction depending on the case.

Preferably, the lower alkyl group consists of 1 to 4, especially 1 or 2 carbon atoms. Aryl represents, for example, a substituted or preferably unsubstituted phenyl group. Heteroaryl preferably represents a heterocyclic group optionally fused to a substituted or unsubstituted benzene ring. An example of a substituent is alkoxy. The cycloalkyl group is preferably a 3- to 10-membered ring, especially a 6-membered ring.
~ Consists of an 8-membered ring. It is preferable that n is an even number, particularly preferably 0.2 or 4.

According to the present invention, 1,2-dithiolane-1-oxygcarboxylic acid- and its pharmaceutically acceptable esters, amides and salts, in particular compounds of the formula (hereinafter referred to as compounds of the invention), have surprisingly In particular, it was found to have immunostimulatory activity.

In particular, the compounds have been found to stimulate not only antigen-reactive lymphoid proliferation, but also antibody production (primary and secondary immune responses) and cell-mediated immune responses.

The immunoactivity of the compounds described above is demonstrated in standard tests both in vitro and in vivo. Positive immunostimulatory activity of the compounds of the invention is demonstrated - for example, by the test method described below.

In vitro: Test ■: Michelle/Du
tton) Test - Induction of humoral response by primary immunization against xenogeneic red blood cells of mouse lung cells cultured in suspension [5
science 153.1004 (1966) and J.
, Exp.

Med, 126, 423 (1967)).

Mouse lung cells are cultured for 3-4 days in the presence of antigen (sheep red blood cells, SE) and test substance. Cells are harvested, washed and plated in semi-solid medium with fresh antigen (SE). After incubating for 60 minutes, complement is added and incubation continues for an additional 90 minutes. During the primary culture period, sensitization of mouse lymphocytes to the antigen releases antibodies. Due to the presence of complement and secreted SE-specific antibodies, sheep red blood cells are lysed (plaque formation). The compound of the present invention was added at a concentration of 0.01 to 10.
Activation of plaque forming cells is observed when used at 0 μf/me.

Test ■; Mixed lymphocyte reaction - CBach et al., J. Exp.
.

Med, 136.1430 (1972)].

reaction (i.e., proliferation and differentiation of lymphoid cells [mouse (Ba1b/c) [1 visceral cells]] co-incubated with allogeneic lung cells obtained from irradiated mice (C:B'A4) for 3 days). Measure in the presence and absence of the test substance. The reaction without the addition of the test substance was used as a control,
Let this be 100%. The response in the presence of the test substance is expressed as a % change compared to the -lOO% control response. Activation of the reaction is observed when the compound of the present invention is used at a concentration of 0.4 to 10 μf/lle −'.

Test ■: T cell specific antigen dinitrophenyl hookhole DN
Three weeks after immunization with P-K LH, mice are boosted with the same antigen. One to four weeks after antigen administration, the lungs are removed and cell cultures are prepared.

A specific antibody expressed in response to the DNI'-KLH antigen is collected from one supernatant and measured by ELISA.

The test substance is added at various concentrations during the incubation of the in vitro cell culture. Antibody expression without test substance is used as a control - this is taken as 100%.

The response in the presence of the test substance is expressed as the % change in response compared to the 100% control response. The compound of the present invention at a concentration of 0.4 to
Reaction (immune response) when used at a concentration of 10μ9/me
activation is observed.

Similar to the above test ①, lymphocytes [mouse (Ba1b/c) spleen cells] in the presence or absence of the test substance.
are co-incubated for 5 days with allogeneic lung cells obtained from irradiated mice (CBA f ). Collect the supernatant,
Amount of immune interferon (L1 infected with virus)
929 fibroblast protection). Interferon units are determined using a human leukocyte interferon preparation of known titer as a standard.

Activation of immune interferon production is achieved by using the compound of the present invention at approximately 0.2 to 5.0 μf/ml! observed in the above test method when used at a concentration of

In vivo: 1) Ietrich et al., Int, Arcb, Aller
gy 38.24fi (1970)].

Antigen (oxacilone) is applied to the abdomen of the mouse to locally sensitize it, and this is designated as day 0. Test compounds are 1 to 5
Administer daily intraperitoneally (i,p) or orally. 9
A challenge (antigen) is performed by applying the antigen to the whites of the eyes and the right ear. After another 24 hours, the skin thickness of both the right ear and the untreated left ear is measured using a microcapilla (micrometer caliper). The average difference in ear thickness between both ears was used as a measure of response evaluation. 0.1 of the compound of the invention daily
Upon oral administration of a dose of ~10 mg/K9 for 5 days i4) or in healthy adult mice with normal immune responses, a significant stimulatory response of a delayed-type hypersensitivity reaction is observed.

et al. “Cell-binding antibodies” (Numa S and Koprow
ski et al.) = Wistar In5t, Pre
ss,, Ph1ladelphia.

U, S, A, pages 109-1221].

Mice were sensitized by intravenously injecting sheep red blood cells (SE),
The test substance is administered i, p on day 0 and month. After 4-10 days,
Kill the mouse-n! Prepare the visceral cell suspension. The cell suspension is plated on semi-solid agar with the new antigen (SE) and incubated for 25 hours in the presence of complement. Sensitized lymphocytes secrete antibodies in response to the antigen and dissolve the antigen (SE) in the presence of complement (plaque formation).

Administration of 0.1-1.0/immediate doses of the compounds of the invention results in an increase in both IgM and IgG antibodies.

The test substance is administered i, p or orally to nude (athymic) mice (Balb/c or C57/BL strain). 16
After an hour, the lungs are removed and the lung cells are incubated with chromium-labeled target cells (YAK-1) for 4 hours. Natural killer cells destroy target cells and release labeled chromium into the supernatant. The supernatant is collected and the amount of released labeled chromium is measured using a scintillation counter. Ill! of untreated mice! - The released amount of labeled cells incubated with visceral cells is used as a control, which is taken as 100%, and the released amount due to test substance administration is expressed as a % change in the released amount compared to the control.

An increase in chromium release is observed and observed by administering 9 doses of test compound from 0.1 to 10.0 q/.

Therefore, the compound represented by formula (I) is an immunostimulant L 1
It was found that it can be used as a systemic immune enhancer and a non-specific host tolerance activator. Thus, a compound of the invention may be used, for example, in the treatment or supportive therapy of conditions associated with impaired immune responses, particularly humoral responses, and/or delayed-type hypersensitivity, and conditions in which an enhancement of the immune response is somehow indicated (i.e. , in combination with other specific or supportive treatments). In particular, the two compounds of the invention have application in the treatment or supportive care of pathological conditions resulting from immune deficiencies found in patients with primary immune deficiencies or geriatric diseases, and in patients with severe burns or systemic infections. The compounds of the invention are also indicated for the treatment and supportive care of viral diseases (for example disseminated herpes - progressive cotton and disseminated varicella) - and Hodgkin's disease and other malignancies.

For the above uses, for the purpose of adjuvant effects such as supportive care, a single oral dose of about 0.1 to about 70 w1 g is applied, and in the latter case divided into 2 to 4 doses per day. It is advantageous to administer the compound of formula (■) in a sustained-release form--and therefore, when daily administration is desired, the indicated unit dosage form for oral administration is from about 0.0025 m9 to about 3.0 m
5 mg, or up to 70 my of a compound of formula (I) if three adjuvant treatments are desired.

Based on the immunostimulatory activity of the compounds of formula (I), they are also applied as adjuvants in vaccines. For the purpose of this application, an administration method is applied in which an oral dose of 0.5 to ~100 my - preferably about 70 mg is administered on the day of vaccination, and if desired, the same dose is readministered as appropriate 2 to 4 weeks later. .

As already mentioned, the suitable daily dosage of a particular compound of the invention depends, among other things, on its relative potency of activity. Preferred compounds in the present invention, namely (anti)-1,2-
Dithiolane-4-carboxylic acid cyclohexylamide-5
-oxide (compound A compared and shown in the primary table).

Compounds of formula (I) in which kl represents a hydroxy or amine group may be used in free form - or in the form of a pharmaceutically acceptable salt, the latter also being equivalent to the free form. It shows some activity.

The compound may be administered enterally, e.g. orally or parenterally.
For example, it can be administered as an injection. Pharmaceutical compositions containing the compounds of this invention are prepared according to standard compounding techniques--eg, by mixing with conventional pharmaceutically acceptable diluents, carriers, or excipients. Such formulations are conveniently formulated - for example, in tablet or capsule form, or in a suitable injection form.

The present invention also provides compounds of the invention which are applied as medicines as mentioned above, in particular as immunostimulants - for example in the treatment or supportive therapy of conditions associated with disorders of the immune response as mentioned above.

The invention further comprises administering an effective amount of a compound of the invention, as described above. Methods of stimulating an immune response in a subject in need of such treatment are provided.

The present invention also provides pharmaceutical compositions comprising a pharmaceutically acceptable diluent or carrier and an inventive compound, as described above.

Among the invention compounds, the compound of formula (1) is preferred.

Of particular interest are compounds of formula (Ip).

Preferred substituents are -R1=a) l) Hlo-alkyl or N■-12
, b) NH-alkyl, c) N-(alkyl)2, d) substituted monoalkylamino, or e) NHCH2(] or 2)-phenyl, cycloalkyl-bicyclic heteroaryl-n==aN even b) O12 or 4, or C)0. Further, preferred positions of COR□ are a) 3 or 5- or b) 4.

A special group of compounds represents a monoalkylamino group in formula (I+3) in which -R1 is substituted with cycloalkyl-phenyl or bicyclic heteroaryl, in which case phenyl or bicyclic heteroaryl is itself substituted with alkoxy It is a compound which is substituted with a group and may be unsubstituted, or a compound where one group is a cycloalkylamine and -0 is 0.

Another group of compounds are those of formula (I) in which kl represents hydroxy, amino, mono- or di-lower alkylamino, lower alkoxy, or MeO, in which case -Me means metal equivalent, and the alkyl group may be substituted with aryl or unsubstituted, and ■ is an integer from 0 to 1o. Among this group of compounds, compounds in which □ is other than hydroxy, MeO1 lower alkoxy or amine are particularly preferred.

In any of the above groups of compounds, compounds in which k□ is in the 4-position are particularly preferred.

The preferred compounds for the above applications are of the (anti) form.

The only particularly preferred compound is (anti)-1°2-dithiolane-4-carboxylic acid cyclomethylamide-5-oxide.

The following examples further illustrate the invention:
All temperatures are expressed in degrees Celsius.

0.37 ml of asparagus acid veratramamide is taken up in 15 ml of dry dichloromethane and cooled to water bath temperature. 15ml of dry dichloromethane! Add 0.21y of 1n-chloroperbenzoic acid dissolved in . After 1 hour, the reaction mixture is shaken with sodium bicarbonate solution, dried over anhydrous magnesium sulphate and concentrated by evaporation.

The crystalline residue is taken up in a small amount of dichloromethane, separated on a medium pressure ram and purified by Merck Lichrosolve.
Li Chro 5orl)) Si (i Q-size C; eluent ethyl diacetate) Syn-isomer: Rf=0.23-mp ~164~
166° anti-isomer: Rf-0,35= mp=
191-193° The following compounds can be obtained in the same manner as in Example 1.

(The "Position" column indicates the bonding position of one side chain; 1t
r-values were determined using toluene/ethyl acetate - 1/1).

The necessary starting materials can be prepared as follows.

a) N-Hydroxysuccinimide ester of asparagus acid 0.32 y of asparagus acid dissolved in 2 Q me of dry dichloromethane, 0.62 F of di(N-succinimide) carbonate, 0.3 me of pyridine and 1 y of anhydrous magnesium sulfate. Add. The mixture is heated to 50° in a nitrogen stream and refluxed for 3 hours with stirring. Next, the reaction mixture is allowed to cool, the magnesium monosulfate is removed, and the Oki liquid is concentrated by evaporation. The residue is purified on a medium pressure ram (Merck Lichrosolve 5i60-size C0 eluent; toluene/ethyl acetate = 1/1; Rf = 0.56), m
, p, 129°.

b) Asparagasic acid veratrylamide 0.062y of N-hydroxysuccinimide ester of asparagasic acid was dissolved in 5me of dry dichloromethane -
To this is added 0.112 minutes 7 of veratrylamine at room temperature. After 20 hours, one reaction mixture is concentrated by evaporation and the residue is purified on a medium pressure ram (Merck Lichrosolve 5i5Q, size A0 eluent; toluene/ethyl acetate = 1/1; Rf = 0.32), in, p .

154° The following starting materials can be obtained in a similar manner (the "Position" column indicates the attachment position of one side chain).

Other compounds which can be used in the present invention are - for example - syn- and anti-1,2-
Dithiolane-3-carboxylic acid-1-oxide (respectively m, P, 115-7° and -148° decomposition λ and syn- and anti-1,2-dithiolane-4-carboxylic acid-1-oxide (respectively m, l ).

130-140° and 130-136°).

Patent Applicant Sand Akchengesellschaft Representative Patent Attorney Aoyama Aoyama and 1 other person (Continued from page 1 209:DO7132-4C 339:00)

Claims (1)

[Claims] +11 A pharmaceutical composition comprising 1,2-dithiolane-1-oxidecarboxylic acid, or a pharmaceutically acceptable ester, amide or salt thereof, and a pharmaceutically acceptable diluent or base. . (2) Formula (■): (wherein, R1 is hydroxy, lower alkoxy, amino,
represents cycloalkylamino- or mono- or di-lower alkylamino, in which each alkyl moiety may be substituted or unsubstituted by aryl-heteroaryl or cycloalkyl, and n is from 0 to 1o.
A pharmaceutical composition comprising a compound in free form or in the form of a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable diluent or base. (3) in formula (I), R1 represents cycloalkylamino, or mono- or di-lower alkylamino - in which each alkyl moiety may be substituted or unsubstituted by aryl, heteroaryl or cycloalkyl; - The pharmaceutical composition according to claim 2, wherein n is an integer of 0 to 10. (4) Formula (Ip): (wherein -kl represents cycloalkylamino- or mono- or di-lower alkylamino, where each alkyl moiety is substituted or unsubstituted by aryl-heteroaryl or cycloalkyl) and n is an integer from θ to 10) in free form or, where possible, in salt form. (5) R' is a) NH-alkyl- b) N-(alkyl)2- C) substituted monoalkylamino- or d) N1-1-CI-12(1 or 2)-phenyl-cycloalkyl, selected from bicyclic heteroaryl and/or the position of the group containing '1 is selected from a) 3 or 5, or b) 4 position - and/or n is a) an even number, b) O=2 or 5. A compound according to claim 4 in free form or, where possible, in salt form, selected from C)0. (6) R'1 represents -cycloalkylphenyl, t or monoalkylamino substituted with bicylic heteroaryl, in which phenyl or bicylic heteroaryl is itself substituted with alkoxy;
or unsubstituted, or in which Ri is cycloalkynogemino and -n is 0, in free form or possibly in salt form. The compound according to any one of claims 4 to 6, wherein (7) has a group containing □ at the 4th position. (8) (Anti)-1,2-dithiolane-4-carboxylic acid cyclohexylmethylamide-1-s-oxide. (9) (Anti)'-1,2-dithiolane-4-carboxylic acid-3,4-dimethoxybenzylamide-1-3-
oxide, (anti)-1,2-dithiolane-4-carboxylic acid benzylamide-1-s-oxide, (anti)
-1,2-dithiolane-4-carboxylic acid cyclohexylamide-1-s-oxide, (anti)-1,2-dithiolane-4-carboxylic acid cyclooctylamide-1-s
-oxide, (anti)-1,2-'; thiolane-4
- A compound selected from carboxylic acid-5-methoxyindol-3-ylamide-1-s-oxide and (anti)-1,2-dithiolane-3-carboxylic acid cyclohexylmethylamide-1-s-oxide. tlO) Claims 4 to 9 which are anti-type
A compound according to any one of paragraphs. 11) A patent consisting of oxidizing a compound of the corresponding formula (Ilp): (wherein -lI□ and n have the meanings given above) and recovering the compound obtained in free form or optionally in salt form. A method for producing the compound and salt thereof according to claim 4. (17J Compound represented by formula (II, A) or (III).