Ascomycin, the method of production thereof and pharmaceutical composition based on them

 

(57) Abstract:

The invention relates to new ascomycin General formula I, where Y represents a phenylene; Z is selected from carboxyl and physiologically hydrolyzable of oxycarbonyl or alkyl, CNS, alkylamino or dialkylamino bearing from 1 to 4 carboxyl or physiologically hydrolyzable oxycarbonyl group; Q is O or S; R1Is H, alkyl or aryl; R2is hydrogen or hydroxyl; R3is methyl, ethyl, propyl or allyl; R4is hydroxyl or alkoxyl; R5-oxoprop or (H, OH), R6- oxoprop, H, HE H, alkoxyl); n is an integer 1 or 2, in free form or in the form of pharmaceutically acceptable salts. The compounds possess anti-inflammatory activity. There is also described a method of production thereof and pharmaceutical composition thereof. 4 C. and 11 C.p. f-crystals, 2 PL.

The invention relates to new ascomycin with pharmaceuticals, such as anti-inflammatory activity. The invention also provides a method of obtaining these new compounds containing pharmaceutical compositions.

Ascomycin, of which the most famous FK-506 and himself Ascomycin, constitute a class of lactam MACR aetsa macrolide immunosuppressant, produced by Streptomyces tsukubaensis N 9993. The structure of FK-506 is known (appendix to the Merck Index, 11 th ed. (1989)) as section A5. Ascomycin is described, for example, U.S. patent 3244592. We also know a large number of related compounds that retain the basic structure of FK-506 and Ascomycin (EP 184162, EP 315973, EP 323042, EP 423714, EP 427680, EP 465426, EP 474126, WO 91/02736, WO 91/13899, WO 91/19495, EP 484936, EP 532088, EP 532089. WO 93/5059). Ascomycin, FK-506 and their structurally similar analogues and derivatives in this specification, all referred to as "ascomycin".

As a class ascomycin are strong immunosuppressive and anti-inflammatory compounds, but their pharmaceutical utility is limited by their toxicity. FK-506, for example, may be toxic to kidneys, liver and Central nervous system in a pharmaceutically suitable doses. In addition, although systemic immunosuppression may be desirable under some conditions, such as graft rejection, it is usually undesirable in the treatment of local inflammatory conditions such as asthma or dermatitis.

It is now established that certain ascomycin carrying one or more than one physiologically hydrolyzable and acceptable oxycarbonyl groups, are highly effective locally active against aemia "plastic" drugs, which means that they are rapidly transformed in vivo to ascomycin carrying unprotected carboxyl group, and these compounds are not systemically active. Additionally openly that the relevant ascomycin bearing unprotected carboxyl groups, although much less active substance than ascomycin carrying physiologically hydrolyzable oxycarbonyl groups, however, are very potent at local, especially skin application. As an acid, and essential forms of ascomycin described here, well tolerated and unlike ascomycin used at the present time, does not cause systemic immunosuppression or noticeable side effects at pharmacologically active doses.

Thus, ascomycin according to this invention represents ascomycin, bearing one or more than one physiologically hydrolyzable and acceptable oxycarbonyl group, or one or more than one carboxyl group, which is applicable for the manufacture of systemically nimmanahaeminda, locally active drugs for local application, for example on the skin or in the respiratory tract, in particular the compound of formula I, Ia or IB, carboxyla grouping or grouping appropriately attached to ascomycin through nametables labile spacer, consisting of a length of from 1-18 carbon units, preferably 6-8 carbon units, and may contain cyclic (for example, aromatic or branched structure and/or one or more than one heteroatom, for example nitrogen, oxygen or sulfur. The spacer preferably contains Allenova grouping, for example fenelonov grouping. When determining the length of spacer elements grouping should be understood that the specified length is the length of the longest consecutive chain of atoms in the group, not counting the side chains or bridge part of cyclic structures. Thus, the length of the p-phenylcarbamoyl of the spacer, for example, can be considered equal to 6 carbon units - four p-phenylene, one for nitrogen and one for the carbonyl. This spacer respectively connected to ascomycin by hydroxyl in the 4-th position tsiklogeksilnogo ring ascomycin (i.e., in the 28th position, using the standard numbering for FK-506), for example by O-urethane or O-thiocarbamates combination. For example, the invention includes 28-O-carbarnoyl - or 28-O - thiocarbamoyl-ascomycin in which carnemolla or thiocarbamoyl grouping is not metabolically labile, but carries or is connected with one of the equipment through a carbon chain, with a length of 16 carbon units, preferably 4-6 carbon units, preferably containing fenelonov grouping. Under "system neomonetarist, locally active" refers to a compound which is at least 10 times more active at the local introduction, than when administered intravenously, in a suitable model of inflammation, for example, at least ten times more active when introduced by inhalation, than when administered intravenously, in sensitized model in Brown-Norway rats, described below.

Under physiologically hydrolyzable and acceptable oxycarbonyl mean a grouping of the formula RO-CO-, which is cleaved under physiological conditions with the formation of (1) alcohol (ROH), portable entered doses, and (2) ascomycin carrying one or more than one carboxyl group. Acceptable oxycarbonyl groups include, for example, alkoxycarbonyl, for example,

(C1-6)alkoxycarbonyl and aryloxyalkyl; preferably methoxycarbonyl, etoxycarbonyl, tert.-butoxycarbonyl or benzyloxycarbonyl.

Ascomycin according to this invention preferably are compounds of the formula I:

< / BR>
where Y is uglevodorov structure, possibly containing up to three heteroatoms, for example nitrogen, oxygen or sulfur and bearing from 1 to 4 carboxyl or physiologically hydrolyzable and acceptable oxycarbonyl groups;

Q is O or S;

R1Is H, alkyl or aryl:

R2is hydrogen or hydroxyl;

R3is methyl, ethyl, propyl or allyl;

R4is hydroxyl or alkoxyl;

R5- oxoprop or (H, OH);

R6- oxoprop, (H, OH) or (H, alkoxyl);

n is 1 or 2; and

the relationship indicated by parallel solid or dashed lines, are either simple or double bond;

provided that when R1is alkyl or aryl, then Y carboxymethyl or physiologically hydrolyzable and acceptable oxycarbonyl)methyl, or aryl, or aryl or alkyl-, alkoxy-, alkylamino or dialkylamino)-aryl, bearing from 1 to 4 carboxyl or physiologically hydrolyzable and acceptable oxycarbonyl groups;

in free form or in the form of a physiologically acceptable salt.

For formula I, independently, preferred the following placeholders:

Y is preferably (1) an alkyl, aryl, alkaryl, alkoxyaryl, aralkyl, Ala or dial-aminoalkyl, or Ala or dial-aminoaryl bearing from 1 to 4 physiologically hydrolyzable and acceptable hydroxy or dial-aminoaryl, bearing from 1 to 4 carboxyl groups. Most preferably, Y carboxymethyl or physiologically hydrolyzable and acceptable oxycarbonyl)methyl, or aryl or alkyl-, alkoxy, alkylamino - or dialkyl-amino)-aryl, bearing from 1 to 4 carboxyl or physiologically hydrolyzable and acceptable oxycarbonyl groups.

Q is preferably O.

R1- preferably H, alkyl or benzyl, for example, H, methyl, benzyl, most preferably H.

R2- preferably hydroxyl.

R3- preferably ethyl or allyl.

R4- preferably hydroxyl.

R5- preferably oxo-group.

R6- preferably (H, methoxy group).

n is preferably 2.

The relationship indicated by parallel solid or dashed lines, is preferably a simple link.

Especially preferred compounds of formula I are the compounds of formula Ia:

< / BR>
in which Q, R1- R6, n and the dotted line such as defined above,

W is methylene or phenylene, and

Z is selected from carboxyl and physiologically hydrolyzable and acceptable oxycarbonyl, and when W - f is or physiologically hydrolyzable and acceptable oxycarbonyl groups

in free form or in the form of pharmaceutically acceptable salts.

Preferably, W is p-phenylene.

Where W is phenylene, such as p-phenylene, Z is preferably selected from benzyloxycarbonylamino, alkoxycarbonyl, (alkoxycarbonyl)1-4of alkyl, (alkoxycarbonylmethyl)1-2amino group, alkoxycarbonyl groups, such as methoxycarbonylmethyl, ethoxycarbonylmethyl, benzyloxycarbonylamino, 2-(methoxycarbonyl)ethyl, 3- (methoxycarbonyl)cut, methoxycarbonyl, 2-(methoxycarbonyl)Attila, ethoxycarbonylmethoxy group, di(methoxycarbonyl)-methyl, di(tert-butoxycarbonyl)-methyl, 2-di(methoxycarbonyl)-ethyl, 1-(diethoxycarbonyl-methyl) -2-di(methoxycarbonyl)-ethyl or di(ethoxycarbonylmethyl)amino group or carboxyl and carboxyethyl, such as carboxymethyl. Where W is methylene, Z is preferably carboxyl or alkoxycarbonyl, such as methoxycarbonyl or etoxycarbonyl.

In particular, a preferred class of compounds are, therefore, the compounds of formula IB:

< / BR>
where Q, R1, R3and Z, as defined above,

in free form or in the form of pharmaceutically acceptable salts.

The most preferred methoxycarbonylmethyl.

It should be understood that the compounds of formulas I, Ia and IB can exist in free form or in some cases can form salts, for example salts obtained by the joining of acid or base at the amino - or carboxyl groups, respectively. It is believed that all these physiologically acceptable form of salts are included in the scope of this invention.

Used here, the terms "ALK" or "alkyl" preferably refers to an aliphatic group containing up to six carbon atoms (chain which may be branched, linear or cyclic, unsaturated possible, possibly interrupted by one or more than one ether (-O-) communication and/or possibly substituted, for example, one or more than one molecule of halogen), especially saturated branched or linear C1-4the alkyl; and the terms "ar" or "aryl" preferably refers to an aromatic hydrocarbon group with a single or double ring, such as phenyl, benzyl, tollu, naphthyl and the like (which may be substituted, for example one or more than one molecule of halogen), especially phenyl or benzyl.

It should be understood that the compounds of formulas I, Ia and IB may exist in F. the practical isomers due to the presence of asymmetric carbon atoms and double bonds, in the form of tautomers thanks enolizatsii or other balanced rearrangements, which are also included in the scope of this invention. The preferred conformation is the same as ascomycin or FK-506, for example of the formula IB.

Compounds of formulas I, Ia and IB are preferably produced by the interaction of 28-O-activated ascomycin (for example the compounds of formula I, where the oxygen in the 4th position collegecollege ring carries an activating group instead carboloy or thiocarbamoyl group) with a compound of formula Y-N(R1)H, where Y and R1such as defined above (in a carboxy-protected form, for example, where Y has one or more carboxy), removing the protection of the product, if necessary, and excretion of compounds of formula I, Ia and IB in free or salt form, 28-O-activated ascomycin respectively produced by the interaction of the source ascomycin, for example the compounds of formula I, having the hydroxyl at the 4-Ohm position tsiklogeksilnogo rings, such as Ascomycin or FK - 506, with O-activating reagent, for example triphosgene in the case of urethane compounds or thiophosgene if THIOCARBAMATE compounds under appropriate conditions of reaction, such as pradov the presence of organic bases. Perhaps the original ascomycin can be hydroxyamine to prevent activation by other localization, in addition to desirable, and protection from the reaction product is removed either before or after the introduction of aminosalicylates, but this operation is not required, especially in the case when activation is carried out at low temperatures, because the hydroxy-group on tsiklogeksilnogo the ring ascomycin, as a rule, much more accessible and reactive, than any other of hydrategraph molecules.

Compounds of formulas I, Ia, IB receive, respectively, as follows:

(1) to obtain compounds of formulas I, Ia and IB, carrying one or more physiologically hydrolyzable and acceptable oxycarbonyl groups, atrificial appropriate ascomycin, bearing one or more carboxyl groups, an appropriate alcohol, such as hydroxyalkyl or hydroxyaryl, in particular methanol, ethanol, tert.-butanol or benzyl alcohol; or

(2) to obtain compounds of formulas I, Ia and IB, carrying one or more carboxyl groups, hydrolyzing the corresponding ascomycin, bearing one or more oxycarbonyl groups

and produce the compound of formula immunodepressive and anti-inflammatory activity. Their peculiarity is the fact that they inhibit antigen-induced infiltration of inflammatory cells, for example, in the respiratory tract or the skin. In vivo this activity manifests itself after local injection, for example, after the local introduction of inhaled through the lungs or application to the skin. A distinctive feature of ascomycin according to the invention is that they exhibit significantly reduced activity or largely devoid of such activity, such as anti-inflammatory or immunosuppressive activity in vivo with the systemic administration, for example oral or intravenous.

Immunosuppressive and anti-inflammatory properties ascomycin according to the invention can be demonstrated in standard test models in vitro and in vivo, for example, as follows:

1. In vitro immunosuppression:

The reaction of the mixed culture of murine lymphocytes

About 0.5 to 106lymphocytes from spleens of females (8-10 weeks) mice of Balb/c incubated for 5 days in 0.2 ml of cell growth medium with 0.5 to 106lymphocytes from spleens of females (8-10 weeks) mice of CBA. The test substance is added to the medium at various concentrations. Activity assessed what about the radioactive thymidine.

Ascomycin of the present invention with a physiologically hydrolyzable oxycarbonyl group, inhibit thymidine incorporation at concentrations of the order of from 0.005 to 0.025 ág/ml of the Compounds of examples 1, 2, 7 and 8 demonstrate the analysis activity, similar or only slightly smaller than the FK-506, with the relative IR50from about 3 to about 15. The acid forms of these compounds, however, are much less active. For example, the compounds of examples 1 and 7 both are transformed in vivo to the acid of example 19, which has a relative IR50equal in this analysis is approximately 150.

2. Model of asthma:

Allergen-induced lung eosinophilia

Effects on rats Brown Norway inhaled antigen (ovalbumin, OA) causes lung eosinophilia after a maximum of 48 hours. In addition to the number of eosinophils, the activation status of these cells can be assessed by measuring the enzymatic activity of the enzyme of the granules eosinophil - peroxidase eosinophils (EPO). In these experiments evaluate the inhibition of the accumulation of pulmonary eosinophils by ascomycin according to this invention.

Ovalbumin (10 μg/ml) is stirred in a mixer for 1 hour on ice) hydroxide alums line Brown Norway (about 200 g). The OA injection with adjuvant repeated after 15 and 21 days. On the 28th day of sensitized rats isolated in plastic tubes and expose aerosol OA (3.2 mg/ml) for one hour, using the system exclusively nasal breathing. Animals are euthanized after 48 hours phenobarbital (250 mg/kg intraperitoneally). Light washed using 3 aliquots (4 ml) in Hanks solution (Hank's) (balanced salt solution HBSS Hanks x 10, 100 ml; EDTA 100 mm, 100 ml; N-2-hydroxyethylpiperazine-N'-2-econsultancy acid (HEPES), 1 M, 10 ml; 1 l of water), the selected cells unite, dried in air by smear and stained for differentiation by types of cells. Cells are identified and counted under oil immersion (x 1000). By counting at least 500 cells in a smear and compute the total population of cells of each type.

In order to determine whether the activity is primarily local or systemic evaluate the relative effectiveness of different techniques. When oral administration of the test substance is administered orally suspended in tragakant, by means of a gastric probe either 1 or 6 hours before and 24 hours after antigen exposure. For intravenous or vorarephilia the introduction of the compound is administered in powder form, suspended in physiological solution containing dimethyl sulfoxide (DMSO) (2,5%). To study the action of a substance through inhalation test substance micronizer for the introduction of experimental animals, fixed in the chamber, adapted for flow of breathing only through the nose. In all cases, the introduction of commit 1 hour or 6 hours before and 24 hours after injection of ovalbumin.

Untreated animals introduction OA induces an increase in all types of cells in BAL (bronchoalveolar lavage) fluid 24 hours after injection. Preliminary introduction of ascomycin in accordance with this invention by inhalation at doses of about 0.1 to 15.0 mg/kg reduces the number of eosinophils in BAL dose-dependent manner compared with control animals not exposed to processing. A number of other leukocytes (macrophages, neutrophils) also decreases. For example, the compounds of examples 1, 2 and 7, below, after intratracheal administration at a dose of about 1 mg/kg or inhalation at a dose of about 0.4 mg/kg demonstrate the ability to inhibit more than 50% accumulation of eosinophils. Unlike FK-506, which is too strong in this model, whether it intravenously, nutrit is almost inactive, or at least 10 times less active when administered intravenously, orally or subcutaneously, thereby demonstrating that their activity is local and not systemic.

3. Skin model:

3.1 Allergic contact dermatitis in the mouse

Estimated anti-inflammatory activity of compounds at the local drawing on models of allergic contact dermatitis in mice. Groups of 8 female mice of NMRI (about 30 g) sensitized by cutaneous application of 2% oxazolone, put a provocative test of oxazolone in the area of the right ear. After 30 minutes the test area treated with topical 10 μl test compound (in the test groups of animals) or vehicle (control group). Left ears are not processed provocative breakdown and do not undergo treatment. The activity determined after 24 hours after application provocative samples by determining the weight of the outer ear as a measure of edema in experimental and control animals. Active compounds inhibit the increase in the weight of the outer ear.

Ascomycin according to this invention, especially those that have one or more than one carboxyl group, demonstrate in this model, the local activity of the same order or slightly above that and FK-506. Ascomycin on this image is ntrace of 0.004 - 0.01% of the compound from example 21, for example, shows from 47% to 69% inhibition compared to 57% inhibition for FK-506 in concentrations of about 0.01%. Ascomycin this invention having one or more than one physiologically hydrolyzable and acceptable oxycarbonyl group, also active, for example, inhibit the swelling of the outer ear by about 35% at concentrations of about 0.01% in the case of compound from example 2.

3.2 Allergic contact dermatitis in domestic pigs

Anti-inflammatory activity entered locally compounds evaluated at the known porcine model of allergic contact dermatitis (J. Invest. Dermatol. (1992) 98: 851-855). Young domestic pigs sensitized with 2,4-dinitrofluorobenzene (DNFB), enter provocative test 1% DNFB on 24 test zones on both dorsolateral sides of the back. The test zone is treated locally dissolved test compounds or only media through 0.5 and 6 hours after administration provocative tests. One day after the introduction of provocative samples semi-quantitatively assess the changes of the areas treated with the test compound and a carrier. Active compounds inhibit erythema and infiltration.

Ascomycin on this retenu, bearing one or more than one carboxyl group, somewhat more potent, for example, at a concentration of 0.13% of the compound from example 20 shows 49% inhibition compared to the areas treated by the media, and the compound from example 2 in the same concentration indicates a 25% inhibition.

Ascomycin according to this invention, accordingly, useful for treatment of diseases or conditions that are sensitive or requiring local anti-inflammatory, immunosuppressive and related therapies, such as local injection in the treatment of diseases or conditions of the eye, nasal passages, buccal cavity or rectum and especially of the skin, respiratory tract or lung. Feature ascomycin this invention is the fact that they allow local anti-inflammatory, immunosuppressive and family therapy, avoiding or reducing unwanted systemic side effects, such as General systemic immunosuppression.

Ascomycin this invention is particularly useful in the treatment by inhalation of diseases and conditions of the Airways or lung, in particular of inflammatory and obstructive diseases is wow, associated with or characterized by infiltration of inflammatory cells or other inflammatory event, accompanied by inflammatory cells, such as accumulation of eosinophils and/or neutrophils. The greatest advantage they have in the treatment of asthma.

Ascomycin according to this invention are useful in the treatment of asthma of any type and origin, including hereditary asthma and acquired bronchial asthma. They are useful in the treatment of atonic and diatopically asthma, asthma induced by exercise, bronchial asthma, including allergic asthma, bronchial asthma induced asthma caused by exercise, manufacturing asthma, asthma induced moved bacterial infection, as well as other non-allergic asthma. Treatment of asthma should be understood as an extensive treatment of the syndrome startrange the baby is breathing", i.e. treatment of the subjects, for example, in the age of 4-5 years, manifesting the symptoms of stridor, especially at night, and diagnosticum as "wheezing infants", which are a separate category of patients that needs close medical supervision and at the present time, more correctly defined as early asthmat the subjects, whose status asthmaticus is steroid-dependent or steroidology.

Ascomycin this invention are also useful for the treatment of bronchitis or for the treatment of chronic or acute respiratory diseases associated with bronchitis. Ascomycin according to this invention can be used for the treatment of bronchitis of any type or origin, including, for example, acute bronchitis, arachidonic bronchitis, catarrhal bronchitis, chronic bronchitis, lobar bronchitis, meningitis tuberculous bronchitis and so forth.

Ascomycin this invention are also useful in the treatment of pneumoconiosis (an inflammatory, widespread disease of the lungs, frequently accompanied by airway obstruction, chronic and acute problems that arise when repeated inhalation of dusts) of any type and origin, including, for example, aluminas, antraks, asbestosis, beryllos, helicos, Philos, sideros, silicosis, tabacos and especially bissines.

Ascomycin according to this invention can also be used to treat associated with eosinophils disorders in the respiratory tract (including pathological infiltration of eosinophils in respiratory tissues) including hypereosinophilia in Utah, resulting from or related syndrome Leffler (Loffler's), eosinophilic pneumonia, parasitic (in particular matatini) infestation (including tropical eosinophilia), bronchopulmonary to aspergillosis, multiple nodular to Takayasu (including syndrome Sharga-Strauss (Churg-Strauss)), the eosinophilic granuloma and eosinophil-related disorders in the respiratory tract, caused by a reaction to medications.

The word "treatment", as used above in connection with the treatment of diseases of the respiratory tract and the lungs, and especially asthma, should be understood as covering both symptomatic and prophylactic treatment, which is an emergency treatment, for example, acute inflammation (symptomatic treatment), as well as advanced treatments for the prevention or limitation of long-term symptomology (prophylactic treatment). The term "treatment" used in the present description and in the claims in connection with these diseases need to be interpreted appropriately, including both symptomatic and prophylactic treatment, for example in the case of asthma symptomatic treatment to relieve acute inflammation and prophylactic treatment to limit SS="ptx2">

Ascomycin according to this invention can also be used to treat any disease or condition of the respiratory tract or lung, requiring immunosuppressive therapy, for example for the treatment of autoimmune diseases of the lungs (for example for the treatment of sarcoidosis, alveolitis or chronic hypersensitive pneumonitis) or to maintain allogeneic transplant lung, for example, if a lung transplant or heart.

As previously described for the purposes specified above, ascomycin according to this invention should be entered locally within the respiratory tract, for example through the lungs and/or by inhalation. Also previously indicated that ascomycin according to this invention is highly effective for local introduction, but no system activity or show a relatively low system activity, for example, by oral administration. Ascomycin according to this invention, therefore, provide a means for treatment of the above diseases and conditions of the Airways or lungs, to avoid undesirable systemic side effects, for example, unintentional ingestion of a drug during inhalation therapy. (Swapimage drugs will be OK to zaputyvaetsja than inhaled).

By applying ascomycin, active in the local introduction of, for example, effective when inhaled, but systemically inactive, this invention makes the treatment ascomycin available for individuals for whom this therapy could be excluded, for example, due to the risk of systemic immunosuppressive side effects.

Ascomycin according to this invention (especially bearing carboxyl group) can also be entered dermal, namely topically on the skin, for example for the treatment of skin diseases mediated by immune mechanisms, such as psoriasis, contact dermatitis, atopic dermatitis, alopecia alopecia, multiple erythema, herpes dermatitis, scleroderma, vitiligo, hypersensitivity vasculitis, urticaria, bullous pemphigoid, lupus, disease, congenital bullous bullosa and other inflammatory and allergic conditions of the skin. Ascomycin according to this invention can be administered in conjunction with anti-inflammatory, immunoagressivnymi or other pharmacologically active agents, such as corticosteroids, antihistamines, antibiotics, antifungal drugs, etc.

Ascomycin according to this invention can be ommanney or inflammatory component, for the treatment of which can be used for local therapy, for example in the treatment of diseases and conditions of the eye such as conjunctivitis, keratoconjunctivitis sicca, and vernal conjunctivitis, as well as to maintain corneal transplant, diseases, acting on the nasal system, including allergic rhinitis, and rectal diseases, such as Crohn's disease (Crohn's) and ulcerative colitis.

For the purposes specified above, ascomycin according to this invention can be applied in any dosage form suitable for local introduction in the desired area.

Thus, for the treatment of diseases of the respiratory tract or lung ascomycin according to this invention can be introduced through the lungs by inhalation using a suitable spray apparatus. For this purpose, ascomycin according to this invention can be produced in any reasonable fine or easily dispersible form suitable for introduction into the respiratory tract or lungs, for example in the form of fine dry particles or in the form of a dispersion or solution in any suitable suitable for introduction through the lungs) solid or liquid carrier. For administration in the form of dry particles ascomycin this is tion materials, in breeding with other suitable powdered inert solid carriers or diluents (such as glucose, lactose, mannitol, sorbitol, ribose, mannose or xylose), in the form of particles with a shell or other suitable form known from the prior art for introducing light through finely ground substances.

Pulmonary administration can be carried out using any suitable system known in the prior art for administration of medicinal substances in dry or liquid form by inhalation, for example aerosol inhaler, atomizer, nebulizer, dry powder, and similar devices. Preferably you should use the device dosed introduction, i.e. a device capable of at each actuation to issue a certain number of ascomycin. Such devices are known from the prior art.

Acceptable forms for local injection for the treatment of diseases and conditions of the skin include, for example, creams, gels, ointments, pastes, poultices, plasters, transdermal stickers, etc. Preparations for application to the skin should contain known from the prior art substance, which improves penetration into the skin, for example on the liquid form through the nasal applicator. The forms used for the treatment of eye, will include lotions, tinctures, gels, ointments and pads for the eyes, known from the prior art. For rectal administration, for example for local therapy of the rectum, ascomycin according to this invention can be administered in the form of a suppository or enema, in particular in solution, such as vegetable oil or similar oil system for use as a retention enema.

The present invention additionally features:

A. a Method of treating diseases or conditions requiring anti-inflammatory, immunosuppressive or family therapy, the subject in need of such treatment, which act locally effective amount of ascomycin according to this invention; and

B. Ascomycin according to this invention for use as a pharmaceutical, e.g. for use in the treatment of diseases and conditions requiring anti-inflammatory, immunosuppressive or family therapy, for example for use in the above method A.

The method described above And is particularly applicable for the treatment of diseases and conditions of the eye, nose, throat, mouth, rectum, or osobennyh next. Ascomycin according to this invention, the bearing physiologically hydrolyzable and acceptable oxycarbonyl group or groups, are particularly suitable for the treatment of diseases or conditions of the Airways or lungs, requiring anti-inflammatory and similar therapies, including graft rejection, lung, and especially any disease or condition of the respiratory tract or lungs, which is characterized by infiltration of inflammatory cells, such as for the treatment of asthma. Ascomycin according to this invention, carrying a carboxy group, or groups, are particularly suitable for the treatment of diseases or skin conditions requiring anti-inflammatory or similar therapy, for example for the treatment of psoriasis, contact dermatitis, atopic dermatitis and other inflammatory or allergic conditions of the skin.

The present invention additionally features:

Century Pharmaceutical composition for local administration, namely in a form suitable for local administration, containing ascomycin according to this invention together with a pharmaceutically acceptable diluent or carrier, or ascomycin according to this invention in the form of, or means, or device, capable of being the firs, mentioned above in In are diluents or carriers acceptable to the local introduction in this field therapy, namely diluents or carriers acceptable to the local insertion through the lungs, skin, nasal, eyes, or rectally. Preparations in the form of local administration, that is capable of carrying out or facilitating local introduction, include, for example, the preparations of the active ingredient (i.e., in this invention) in the form of dry powder in substantially pure form, for example, such known from the prior art drugs used for administration of the inhalation device for dry powder. Medium or device capable of carrying out or facilitating local introduction, include devices such as inhalers and containers and similar means by which the active ingredients can be delivered in a format suitable for local application. The preferred embodiments according to the paragraph are the following: (1) providing the opportunity for local injection into the respiratory tract or lungs, for example by inhalatio, in the case of ascomycin according to this invention, bearing one or more than one oxycarbonyl grouping, and (2) providing in the appropriate one or more than one carboxyl group.

Dose ascomycin according to this invention used for the implementation of the method according to this invention will, of course, vary depending on the places intended for treatment of a particular state, intended for the treatment, the severity of the condition, the subject to be treated (for example, relative to its weight, age and the like), and the desired effect.

In General, for the treatment of diseases or conditions of the Airways or lungs, for example when used in the treatment of inflammatory or obstructive diseases of the Airways such as asthma, ascomycin according to this invention will be administered locally in the respiratory tract or lungs, for example, by inhalation, in doses of about 0.01-50 mg per day, such as 0.1-5 mg / day, most preferably from 0.4 to 1.6 mg/day, for example, if the introduction of the system dosed introduction to the series from 1 to 5 breaths each time with the introduction of repeated introductions from 1 to 4 times a day, for example, 200 to 800 mg once or twice daily by inhalation. The dose at each introduction will therefore be approximately 0,0025 - 10 mg, more acceptable to 0.1 - 1.0 mg, for example, with the introduction of the system has dosed the introduction, in particular on ascomycin according to this invention will, basically, be entered in the form of appropriate compositions, for example, eye drops, gel, lotion for the eyes, etc. or nasal drops, aerosol for nasal or so on, when the content of 0.005 to about 5%, especially from about 0.01 to about 1% of ascomycin by weight, in the corresponding acceptable for ophthalmic or nasal introduction of the diluent or carrier for administration to the surface of the eye or nasal in the amount of from about 0.05 to about 0.2 ml of the compositions, for example from about 0.05 to about 0.1 ml of the composition, one or two to three times daily.

For the treatment of diseases or conditions of the rectum in General, an acceptable daily dose of ascomycin according to the invention will be of the order of from about 0.01 to about 5, preferably from about 0.1 to about 1.0 mg/kg when injected into the form of a retention enema once or in divided doses 2x a day. Each entered the dose will be, thus, respectively, contain from about 0.1 to about 350, preferably from about 1 to about 150, or more preferably from about 5 to about 70 mg ascomycin according to this invention together with suitable for rectal injection diluent or carrier. Acceptable concentration osCommerce 2, preferably from about 0.1 to about 1.0 mg/ml

For dermal injection in the treatment of diseases and conditions skin ascomycin according to this invention will be entered into primarily acceptable for application to the skin form containing a therapeutically effective concentration of ascomycin according to this invention, for example from about 0.001 to 10%, for example 0,004% - 1% by weight of ascomycin according to this invention together with acceptable for application to the skin diluent or carrier. Preparations for dermal injection can be in the form of creams, ointments, gels and systems for percutaneous introduction, for example patches, and in addition to inert diluents or carriers may also contain appropriate agents that improve the penetration into the skin, like the compositions known from the prior art. Such compositions will be applied to land for treatment, in the amount of roughly 0.005 to about 0.05 g/cm2, 1, 2 or 3 times a day.

EXAMPLES

Table I compares the compounds of formula IB in which Z, Q, R1and R3are as specified. Bz means benzyl.

Table II provides additional examples of compounds of the formula I, in which R2and R4- the Noah and straight lines, is a simple link, a, Y, Q, R1and R3such as specified. The stereochemistry of all compounds refers to the formula IB.

Example 1 is prepared as follows.

To a solution of Ascomycin (6.0 g, EUR 7.57 mmol) and 4-dimethylaminopyridine (4,62 g, 37.8 mmol) in 75 ml of dichloromethane is added dropwise a solution of triphosgene (0.84 g, and 2.83 mmol) in dichloromethane (15 ml) at -77oC. After 1 hour, add solid dimethyl ester 2-(4-AMINOPHENYL)-malonic acid (2.7 g, 12.1 mmol). The cooling bath is removed and the slurry is heated to the ambient temperature. The reaction mixture is stirred for another hour and add ethyl acetate and a saturated aqueous solution of NaCl. Then the organic phase is twice washed (1N. HCl) and evaporated to dryness, getting foamy residue, after purification with flash chromatography gives pure product, so pl. 115-120oC (ethanol-water), (M + Li)+= 1047.

Example 2 is prepared as follows.

To a solution of FK506 (10.0 g, 12.4 mmol) and 4-dimethylaminopyridine (7.5 g, of 61.7 mmol) in 60 ml of dichloromethane is added dropwise a solution of triphosgene (1,36 r, 4.6 mmol) in dichloromethane (40 ml) at -77oC. After one hour, add methyl ester 4-aminophenylarsonic acid (3.1 g, 18.5 mmol). The cooling bath is removed and samlaut ethyl acetate and a saturated aqueous solution of NaCl. Then the organic phase is twice washed (1N. HCl) and evaporated to dryness, getting foamy residue, after purification by the method of flash-chromatography gives pure product, so pl. 105-109oC (ethanol-water); (M+Li)+= 1001;13C-NMR(CDCl3) selected data: 212,79, 298,10, 172,03, 153,04, 137,03, 129,76, 119,75, 96,90, 51,94, 40,41, 39,57, 9,30.

Examples 3-18 and 21-26 prepared similarly, using as source material Ascomycin, where R3- ethyl, and FK506, where R3-allyl, and replacing the corresponding molar amount of an amine of the formula X-Y'-N(R')H (X, Y' and R' are the same as described above) in dimethyl ester 2-(4-AMINOPHENYL)-malonic acid from example 1 or the methyl ester of 4-aminophenylarsonic acid from example 2. Thiocarbamide connection (for example, where Q - S) are prepared similarly, using thiophosgene instead of triphosgene as aktivirujushchej reagent.

Example 19 was prepared as follows. To a solution of Ascomycin (6.0 g, EUR 7.57 mmol) and 4-dimethylaminopyridine (4,62 g, 37.8 mmol) in 75 ml dichloromethane added dropwise a solution of triphosgene (0.84 g, and 2.83 mmol) in dichloromethane (15 ml) at -77oC. After one hour, add trimethylsilyloxy ether 4-amino-phenylacetic acid (12.1 mmol). The cooling bath is removed, and the slurry is heated aqueous solution of NaCl. Then the organic phase is twice washed (1N. HCl) and evaporated to dryness, obtaining a residue, which is purified by the method of flash chromatography. The resulting product can be further purified by precipitation from a mixture of ethanol-water. The purified compound has a melting point 132-135oC.

Example 20 is prepared as example 19, using FK-506 as a starting material instead of ascomycin in order to obtain a pure compound with a melting point 135-138oC.

1. Ascomycin formula I

< / BR>
where Y represents a phenylene;

Z is selected from carboxyl, physiologically hydrolyzable and acceptable oxycarbonyl, or from alkyl, CNS, alkylamino or dialkylamino bearing from 1 to 4 carboxyl or physiologically hydrolyzable oxycarbonyl groups;

Q represents O or S;

R1represents H, alkyl or aryl;

R2represents hydrogen or hydroxyl;

R3represents methyl, ethyl, propyl or allyl;

R4represents a hydroxyl or alkoxyl;

R5is oxoprop or (H, HE);

R6is oxoprop, (H, OH) or (H, alkoxyl);

is made or a simple or double bond,

in free form or in the form of pharmaceutically acceptable salts.

2. Connection on p. 1 of formula IB

< / BR>
where Q, R1and R3such, as defined in paragraph 1 for formula I;

Z represents carboxyl or physiologically hydrolyzable and acceptable oxycarbonyl or alkyl, alkoxy, alkylamino or dialkyl-amino group bearing one to four carboxyl or physiologically hydrolyzable oxycarbonyl groups

in free form or in the form of pharmaceutically acceptable salts.

3. Connection on p. 2 of formula IB, where Q represents O; R1represents H; R3represents ethyl or allyl; Z represents a carboxymethyl or methoxycarbonylmethyl.

4. The compound according to any one of paragraphs.1 to 3, bearing from one to four carboxyl groups.

5. The compound according to any one of paragraphs.1 to 3, bearing from one to four physiologically hydrolyzable and acceptable oxycarbonyl groups.

6. Connection on p. 3, where R3represents allyl, and Z is methoxycarbonylmethyl.

7. Connection on p. 3, where R3represents allyl, and Z represents the immunosuppressive anti-inflammatory agent.

9. Ascomycin formula I, bearing one or more than one physiologically hydrolyzable and acceptable oxycarbonyl group or one or more than one carboxyl group, PP.1 to 7 for the manufacture of the system nimmanahaeminda local active drugs for local application.

10. Ascomycin under item 9, which is a connection on p. 4 or 7, for the manufacture of a medicinal product for the treatment of inflammatory or autoimmune diseases of the skin.

11. Ascomycin under item 9, which is a connection on p. 5 or 6, for the manufacture of a medicinal product for the treatment of asthma.

12. Pharmaceutical compositions with local anti-inflammatory activity, not with systemic immunosuppressive properties containing the compound according to any one of paragraphs.1 - 7, possibly in combination or in combination with a pharmaceutically acceptable diluent or carrier.

13. Pharmaceutical composition for p. 12 containing compound under item 4 or 7, for the treatment of inflammatory or autoimmune diseases of the skin by topical application.

14. Pharmaceutical composition for p. 12 containing compound under item 5 or 6, the treatment is different in that, that (1) 28-About-activated ascomycin obtained by the interaction of ascomycin with triphosgene or thiophosgene, is subjected to the interaction with the compound of the formula Z-Y-N(R1)N, in which Z, Y, R1are specified in the PP. 1 - 7, possibly in carboxyterminal form, and, if necessary, followed by removal of the protection of the product, or (2) to obtain a connection on p. 5 atrificial corresponding connection on p. 4 the corresponding alcohol, or (3) to obtain a connection on p. 4 hydrolyzing the corresponding connection on p. 5, and excrete the compound of formula I and IB in free form or in salt form.

Priority points:

06.04.1995 and 20.12.1995 - the invention is equally disclosed in the first application corresponding to the specified priorities.

 

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Esters of rapamycin // 2160739

The invention relates to a derivative of rapamycin, method of production thereof, their use as pharmaceuticals and pharmaceutical compositions containing them

The invention relates to the field of synthesis of therapeutically active product

The invention relates to previously unknown compounds, useful in medical and veterinary practice, to their pharmaceutically acceptable salts and biopremier derivatives, to methods for obtaining data of new compounds, to pharmaceutical compositions containing these new compounds, to a single dosage forms of these compositions and to methods of treating patients using these compositions and dosage forms

The invention relates to medicine, specifically to medicines, exhibiting anti-allergic, anti-asthma and anti-inflammatory effect

The invention relates to the field of medicine and is suitable for the treatment of seasonal and perennial allergic rhinitis, conjunctivitis, hay fever, urticaria, including chronic idiopathic urticaria, angioedema, psevdoallergicakie reactions caused by the release of histamine, itching dermatoses, allergic reactions to insect bites and itching of various etiology

The invention relates to cyclopentapeptide formula (I):

cyclo (A-B-C-E-F-(D)-Ala)

in which A, B, C, E and F, independently of one another, may be the same or different and represent a residue of a natural amino acid except cysteine (Cys) and tryptophan (Trp), and accordingly can be an alanine (Ala), arginine (Arg), asparagine (Asn), aspartic acid (Asp), glutamine (Gln), glutamic acid (Glu), glycine (Gly), histidine (His), isoleucine (Ile), leucine (Leu), lysine (Lys), methionine (Met), phenylalanine (Phe), Proline (Pro), serine (Ser), tryptophan (Thr), tyrosine (Tyr) or valine (Val), as well as their physiologically compatible salts

The invention relates to new derivatives of asola General formula I, where R1and R2the same or different, each represents hydrogen, cycloalkyl and so forth, or R1and R2forming (a) a condensed ring, (b) or (C), which may be optionally substituted substituted lower alkyl, amino group and the like; R3, R6, R7, R8the same or different, each represents a hydrogen atom, and so on; R4represents a cyano, tetrazolyl, -COOR9and so on; R5represents a hydrogen atom or lower alkyl; D represents optionally substituted lower alkylene; X and Z are the same or different, each represents oxygen or sulfur, Y is-N= or-CH=; A is-B is-O-, -S-B-, -B-S - or-In-; represents the lowest alkylene or lower albaniles; n = 2

The invention relates to medicine, in particular to pharmacology concerns anti-allergic and anti-inflammatory agents of the formula I, with low toxicity

The invention relates to the medical industry, namely the production of medicines, containing four, pathcomponent mixture of acetylsalicylic acid, paracetamol, ascorbic acid, organic salts of calcium, Dimedrol, and(or) routine used for relief of acute manifestations of influenza
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