Classifications

• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07J—STEROIDS
  • C07J43/00—Normal steroids having a nitrogen-containing hetero ring spiro-condensed or not condensed with the cyclopenta(a)hydrophenanthrene skeleton
  • C07J43/003—Normal steroids having a nitrogen-containing hetero ring spiro-condensed or not condensed with the cyclopenta(a)hydrophenanthrene skeleton not condensed
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
  • A61P1/04—Drugs for disorders of the alimentary tract or the digestive system for ulcers, gastritis or reflux esophagitis, e.g. antacids, inhibitors of acid secretion, mucosal protectants
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P11/00—Drugs for disorders of the respiratory system
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P11/00—Drugs for disorders of the respiratory system
  • A61P11/06—Antiasthmatics
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P11/00—Drugs for disorders of the respiratory system
  • A61P11/08—Bronchodilators
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P13/00—Drugs for disorders of the urinary system
  • A61P13/12—Drugs for disorders of the urinary system of the kidneys
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P15/00—Drugs for genital or sexual disorders; Contraceptives
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P17/00—Drugs for dermatological disorders
• • A—HUMAN NECESSITIES
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  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P19/00—Drugs for skeletal disorders
  • A61P19/02—Drugs for skeletal disorders for joint disorders, e.g. arthritis, arthrosis
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P25/00—Drugs for disorders of the nervous system
  • A61P25/02—Drugs for disorders of the nervous system for peripheral neuropathies
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P25/00—Drugs for disorders of the nervous system
  • A61P25/28—Drugs for disorders of the nervous system for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P27/00—Drugs for disorders of the senses
  • A61P27/02—Ophthalmic agents
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P35/00—Antineoplastic agents
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P37/00—Drugs for immunological or allergic disorders
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P37/00—Drugs for immunological or allergic disorders
  • A61P37/02—Immunomodulators
  • A61P37/06—Immunosuppressants, e.g. drugs for graft rejection
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P5/00—Drugs for disorders of the endocrine system
  • A61P5/24—Drugs for disorders of the endocrine system of the sex hormones
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P5/00—Drugs for disorders of the endocrine system
  • A61P5/38—Drugs for disorders of the endocrine system of the suprarenal hormones
  • A61P5/44—Glucocorticosteroids; Drugs increasing or potentiating the activity of glucocorticosteroids
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P9/00—Drugs for disorders of the cardiovascular system
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07J—STEROIDS
  • C07J41/00—Normal steroids containing one or more nitrogen atoms not belonging to a hetero ring
  • C07J41/0033—Normal steroids containing one or more nitrogen atoms not belonging to a hetero ring not covered by C07J41/0005
  • C07J41/005—Normal steroids containing one or more nitrogen atoms not belonging to a hetero ring not covered by C07J41/0005 the 17-beta position being substituted by an uninterrupted chain of only two carbon atoms, e.g. pregnane derivatives

Definitions

• the present invention relates to steroidal compounds having an improved pharmacological activity and lower side effects and an improved receptor affinity on the specific receptors of endogenous steroids.
• the invention relates to steroidal compounds having an improved receptor affinity on the specific receptors of the endogenous steroids and having an improved pharmacological activity and lower side effects, in particular: those affecting the bony tissue, such for example osteoporosis, osteonecrosis and myopathies, which in patients affected by asthma or by COPD (Chronic Obstructive Pulmu- nary Disease) can determine a remarkable reduction of the respiratory activity; those affecting the gastrointestinal apparatus.
• COPD Choronic Obstructive Pulmu- nary Disease
• the invention relates to compounds having a steroidal structure in particular having not only an improved antiin- flammatory activity at peripheral level, but also an improved anti-neurodegenerative activity, an improved antiarthritic acitivity, an improved immunodepressive activity, an improved angiostatic/angiogenetic and antiasthmatic activity; or usable in substitutive hormonal therapies, for example in the post- menopause therapy.
• the present invention relates also to steroid compounds of the glucocorticoid class which can be used as bronchodilators in respiratory pathologies characterized by broncho-constrictive events.
• the compounds according to the present invention are therapeutically useful in the treatment of illnesses wherein steroidal products are generally applied, with increased benefit, in terms of improved tolerability as above defined and improved efficacy.
• the present invention products give a combination of results, considered as improvement of the therapeutic performance, i.e. improved pharmacotherapeutic efficacy and improved tolerability, compared with the prior art products.
• the present invention products are characterized in that they show an improved therapeutic profile: high activity in the above applications combined with lower side effects as above defined.
• steroids comprise: corticosteroids, classified in glucocorticoids active on the glucogenesis and on the metabolism of proteins, lipids, carbohydrates and calcium in general, mineralcorti- coids active on the water and saline balance; sexual steroids, including estrogens and androgens . It is well known that glucocorticoids represent a first choice pharmacological approach in the therapy of various pathologies.
• Said class of drugs among which, for example, hy- drocortisone, cortisone, prednisone, prednisolone, fludrocor- tisone, desoxycorticosterone, methylprednisolone, tria- mcinolone, paramethasone, betamethasone, dexamethasone, tri- amcinolone acetonide, fluocinolone acetonide, beclomethasone, acetoxypregnelone, etc. can be mentioned, exerts marked phar- maco-toxicological effects on various organs. For said reason the prolonged clinic use and the interruption of the pharmacological treatment cause side effects, some of them very serious.
• nitrooxy derivatives of steroids which are usable also as cardiovascular agents for the coronary insufficiency or angina pectoris therapy, are described.
• German patent DE 2,222,491 describes the preparation of pregnane derivatives having in position 21 the -CH 2 -0-N0 2 group.
• said derivatives have a cardiotropic activity. This activity represents a drawback for said compounds, since they modify the cardiac frequency.
• USP 3,494,941 describes steroid derivatives from 3- hydroxy-extrane or from extr-4 en-3 one, used as vasodilators in the treatment of cardiac affections such as coronary insufficiency and angina pectoris.
• a 0N0 2 group is at the free end of the alkylene chain which is linked by an ether bond to the steroid in position 17.
• nitrate groups also in the positions 3 and 16 of the steroidal structure.
• the same drawbacks mentioned above as regards the effects on the cardiac frequency can be repeated for the compounds of this patent.
• WO 98/15568 in the name of the Applicant describes nitrate esters of steroidal compounds, wherein between the steroidal structure and the nitrooxy group a bivalent linking group is inserted. Said compounds show a good efficacy and/or good tolerability with respect to the corresponding precursors. However in the examples and in the description no data are reported on the receptor activity. No indication is therefore reported suggesting steroidal compounds having an improved receptor activity and an improved pharmacological activity combined with lower side effects.
• Patent application WO 00/61604 in the name of the Applicant describes nitrooxy derivatives of steroidal compounds with various linking groups having at one end a nitrooxy group, and covalently linked with the other end to a steroidal compound.
• the uses concern the compounds usable in the treatment of patients in oxidative stress.
• Said compounds contain in the molecule also a bivalent linking group which must be capable to prevent the free radicals production and is selected on the basis of the tests reported therein. No indication is therefore given suggesting steroidal compounds having an improved receptorial activity and an improved pharmacological activity combined with lower side effects .
• the Applicant has surprisingly and unexpectedly found a specific class of steroidal compounds which in the above pathologies unexpectedly and surprisingly show not only an improved efficacy but also an improved tolerability and lower side effects compared with the steroids of the prior art .
• An object of the present invention are nitrooxy derivatives of steroidal compounds of general formula
• B is a steroidal radical having the following structure:
• RAi is H, CH 3 ;
• RA 2 is a C ⁇ C ⁇ 0 linear or branched alkyl chain, preferably C ⁇ -C 3 , still more preferably CH 3 , or a saturated cycloaliphatic ring having 5-6 carbon atoms or an aromatic ring optionally substituted in para position with N(Ric)2 wherein R lc is a C ⁇ -C 10 , preferably C ⁇ C 4 , linear or branched alkyl;
• R" in position 17 is a bivalent radical having one of the following meanings :
• na, n'a, and n''a, equal to or different from each other are integers from 0 to 6, preferably 1-3; nb, n'b, n' 'b and n' ' 'b, equal to or different from each other, are integers equal to to 0 or 1; R 4 , R 5 , R 4 » , Rs- , R 4 » , R 5" , equal to or different from each other, are selected from H, C 1 -C 5 , preferably C1-C 3 linear or branched alkyl;
• Xx is a bivalent linking group selected from the following: ⁇ YARI• '
• nIX is an integer from 0 to 10, preferably 1-3; nllX is an integer from 1 to 10, preferably 1-5; RTIX, RTIX', RT IIX , R TI IX 1 ⁇ equal to or different from each other are H or C 1 -C 4 linear or branched alkyl; preferably RTIX, RTIX RTIIX, RTIIX' 3 ⁇ 6 H;
• Y 3 is a saturated, unsaturated or aromatic heterocyclic ring, having 5 or 6 atoms, containing from one to three heteroatoms, preferably from one to two, said heteroatoms being equal or different and selected from nitrogen, oxygen, sulphur; preferably nitrogen; t3 is zero or 1; Z has the following meaning:
• T has the following meanings:
• n3 and n' 3 are as above defined.
• the linking group X x links to the radical B with the indicated valence which does not bring the oxygen.
• Y 3 is selected from the following bivalent radicals :
• Y 3 (Y12), having the two free valences in the ortho positions with respect to the nitrogen atom; (Y16) with the two valences linked to the two heteroatoms, (Yl) (pyrazol) 3, 5-disubstituted; (Y16) is particularly preferred.
• the invention preferred compounds are those wherein the precursor of B has the meanings mentioned below.
• precursors of the steroids of the present invention can be mentioned those described in the Merck Index, 12th Ed. 1996, herein integrally incorporated by reference, in which also the respective synthesis methods are mentioned, and in the patents indicated hereinafter.
• the precursors are the following: corticoster- oids selected from Budesonide, Hydrocortisone, Alclomethasone, Algestone, Beclo ethasone, Betamethasone, Chloroprednisone, Ciclesonide (USP 5,482,934), Clobetasol, Clobetasone, Clocor- tolone, Cloprednol, Cortisone, Corticosterone, Deflazacort, Desonide, Desoximethasone, Dexamethasone, Dexamethasone 17- furoate, Diflorasone Diflucortolone, Difluprednate, Flu- azacort, Flucloronide, Flumethasone,
• R" -0-, wherein the free valence of the oxygen is saturated with H;
• H in the formula (IA) is substituted with a group different from OH.
• the precursors of the bivalent radicals X x as above defined, wherein the oxygen free valence is saturated with H and the free valence of the end carbon atom is saturated either with a carboxylic or hydroxyl or amminic group, are commercial products or they can be synthesized according to known methods of the prior art .
• the preferred compounds according to the present invention are the following: Hydrocortisone 21- (4' -nitrooxymethyl) benzoate
• Dexamethasone-21 [2- [4- [2- [4- (nitrooxymethyl) benzoyloxy] ethyl] 1-piperazinyl] acetate]
• Flumethasone-21 [2- [4- [2- [4- (nitrooxymethyl) benzoyloxy] ethyl] 1-piperazinyl] acetate]
• connection between B and Xi is, as said, of ester or amidic type (NH or NR 1C , as defined in X 0 ) .
• ester or amidic type NH or NR 1C , as defined in X 0
• synthesis methods de- scribed in the prior art are usable.
• the compounds according to the present invention when at least a functional group salifiable with acids, for example an amminic group, is present, can be transformed into the corresponding salts.
• one way to form the salts is the following: when one basic nitrogen atom is present in the molecule, it is reacted in organic solvent such for example acetonitrile, tetrahydrofuran, with an equimolecular amount of the corresponding organic or inorganic acid.
• organic acids are: oxalic, tartaric, maleic, succinic, citric, tri luoroacetic acid.
• inorganic acids are: nitric, hydrochloric, sulphuric, phosphoric acid.
• the precursor compounds usable in the present invention have one or more chiral cores, they can be in a racemic form or as diastereosisomer mixtures, as single enantiomers or single diastereoisomers; if they show a geometric asymmetry the compounds can be used in the cis or trans form.
• acylhalides used in the invention compound synthesis can be prepared according to known methods of the prior art, for example by reacting the corresponding carboxylic acids with thionyl chloride or oxalyl chloride, with P 111 or P v hal- ides in solvents inert under the reaction conditions. 1. If the steroid reactive function is the hydroxyl group (B- OH) and the bond between the steroid and the linking group Xi is of the ester type, the most used synthesis methods are the following: la.
• the corresponding nitrooxy derivative is obtained by reacting the compound (IA.l) obtained from the previous reaction with AgN0 3 in an organic solvent as acetonitrile, tetrahydrofuran at a temperature in the range 25°C-80°C, according to the following scheme (IA.2):
• the compound HO-C (0) -X 1A -Hal wherein Hal and X XA have the above meanings can be treated with a carboxyl activating agent, selected from N,N-dicarbonyldiimidazol (CDI), N-hydroxy benzo- triazol or dicyclohexylcarbodiimide (DCC) , in an organic solvent such for example DMF, tetrahydrofuran, chloroform, etc., at a temperature in the range -5 and 50 °C.
• a carboxyl activating agent selected from N,N-dicarbonyldiimidazol (CDI), N-hydroxy benzo- triazol or dicyclohexylcarbodiimide (DCC)
• CDI N,N-dicarbonyldiimidazol
• DCC dicyclohexylcarbodiimide
• the obtained compound is reacted in situ with the steroid (B-OH) to give the compound of formula
• the usable synthesis method is for example the following:
• the steroid reactive function is the carboxyl group (R- COOH) and the bond between the steroid and the linking group Xi is of ester type, the most used synthesis method is the following:
• the steroid (R-COOH) is treated with an agent activating the carboxyl selected from N, N-dicarbonyldiimidazol (CDI), N- hydroxybenzotriazol or dicyclohexylcarbodiimide (DCC) in an organic solvent such for example DMF, tetrahydrofuran, chloroform, etc., at a temperature in the range -5°C-50°C.
• CDI N, N-dicarbonyldiimidazol
• DCC dicyclohexylcarbodiimide
• the obtained compound is reacted in situ with the precursor of Xi of formula HO-X ⁇ A -Hal wherein X iA is a radical obtained from Y AR ⁇ or Y P omitting the oxygen atom -0-, and Hal is as above defined.
• the obtained compound, having general formula B-C (0) -0- X ⁇ A -Hal is reacted with AgN0 3 as above described -
• the steroid reactive function is the carboxyl group (R- COOH) and the bond between the steroid and the linking group Xi is of amidic type, the most used synthesis method is the following:
• the steroid (R-COOH) is treated with an agent activating the carboxyl selected from dicyclohexylcarbodiimide (DCC) in an organic solvent as for example DMF, tetrahydrofuran, chloroform, etc., at a temperature in the range -5° and 50°C and the obtained compound is reacted in situ with the precursor of Xi of formula H 2 N-X ⁇ A -Hal wherein X iA and Hal are as defined above.
• DCC dicyclohexylcarbodiimide
• organic solvent as for example DMF, tetrahydrofuran, chloroform, etc.
• the invention compounds wherein the linking group Xi is selected from the above mentioned bivalent radicals, allow to obtain, see the examples of the receptor binding assays, results unexpectedly and surprisingly improved with respect to the nitrooxy derivatives wherein the linking group Xi is an alkylene and/or with respect to the corresponding precursor steroids.
• the present invention compounds do not affect the cardiocirculatory parameters and therefore the present invention compounds do not give undesired effects on the systemic pressure and on the cardiac frequency.
• the invention compounds show an improved pharmacological activity combined with lower side effects, in particular: affecting the bony tissue, such for example osteroporo- sis; affecting the gastrointestinal apparatus.
• the invention compounds have not only an improved antiin- flammatory activity -at a peripheral level, but also an improved anti-neurodegenerative activity, the compounds being active on the neurodegenerative diseases on an inflammatory and traumatic basis of the nervous system, such for example spinal trauma and lesions and cerebral trauma, inflammation of the nervous tracts such as the sclerosis multipla.
• the invention compounds show furthermore an improved antiarthritic activity, improved immunodepressive activity, improved angi- ostatic/angiogenetic and antiasthmatic activity.
• the invention compounds are usable in substitutive hormonal therapies, for example in the post-menopause therapy.
• the compounds according to the present invention are therapeutically useful in the treatment of morbid conditions wherein steroidal precursor products are used, but with increased benefit, in terms of improved tolerability as defined above and improved efficacy.
• the present invention products are characterized in that they show an improved therapeutic profile: high activity in the above applications combined with lower side effects as defined above.
• the invention compounds show lower side effects, in particular as regards : those affecting the bony tissue, such for example osteoporosis, osteonecrosis and myopathies, which in patients affected by asthma or by COPD (Chronic Obstructive Pulmu- nary Disease) can determine a remarkable reduction of the respiratory capabilities; those affecting the cardiovascular system which generate hypertensive responses and/or cardiac frequency diseases; lower predisposition to infections; those affecting the gastrointestinal apparatus.
• COPD Choronic Obstructive Pulmu- nary Disease
• the compounds object of the present invention are formulated in the corresponding pharmaceutical compositions, also with belated release, for parenteral, oral and topic use, such as for example sublingual, inhalatory, suppository,- transdermal, enema, according to the well known techniques in the art, together with the usual excipients; see for example the publication "Remington's Pharmaceutical Sciences” 15th Ed.
• the amount on a molar basis of the active principle in said compositions is generally the same, or lower than that of the corresponding precursor drug.
• the daily administrable doses are those of the precursor drugs, or optionally lower.
• the precursor daily doses can be found in the publications of the field, such for example in the "Physician's Desk reference".
• the present invention compounds are used for the treatment of pathologies wherein the precursor steroids are used.
• the use is mentioned as drugs in rheumatic diseases, renal and bronchial pathologies, ocular and dermatolog- ical diseases, autoimmune diseases, tumoral processes, also in combination with chemotherapeutic and/or radiotherapeutic treatments, in neurodegenerative diseases, for example in spinal lesions from trauma and in the post-transplant therapy.
• chemotherapeutic and/or radiotherapeutic treatments in neurodegenerative diseases, for example in spinal lesions from trauma and in the post-transplant therapy.
• gatrointes- tinal system Crohn disease, ulcerous colitis and IBD (inflammatory bowel diseases) can be mentioned.
• the Applicant has surprisingly and unexpectedly found that the invention steroids of the glucocorticoid class can be used, differently from precursors, in respiratory pathologies characterized by broncho-obstructive events. Said fact is quite unexpected since the precursors are substantially ineffective under said morbid conditions; indeed they must be associated with broncho-dilators as beta-agonists such for example salbutamol.
• the Applicant has found that not only the nitrooxyderivatives of the steroids according to the present invention are effective, but also the derivatives in which the linking group Xi in formula (I) is an aliphatic linking group of the glucocorticoid class of formula (I), selected from the following:
• R'O An alkylenoxy group R'O wherein R' is C ⁇ -C 20 '- linear or branched when possible, preferably having from 2 to 6 carbon atoms, or a cycloalkylene having from 5 to 7 carbon atoms, in the cycloalkylene ring one or more carbon atoms can be substituted by heteroatoms, the ring can have side chains of R' type, R' being as above defined;
• nf is an integer from 1 to 6 preferably from 1 to 4;
• R ⁇ f H, CH 3 and nf is an integer from 1 to 6; preferably from 1 to 4.
• the present invention compounds, differently from the precursors, have no side effects on the bony system, in particular they do not cause bony reabsorption, besides they show a high gastric tolerability.
• N-t-butoxycarbonylpiperazine (3 mmoles, 558 mg) (prepared according to the procedure described by Boschi D. et Al . Arch. Pharm. 1994, 327, 661-667) is dissolved in 15 ml of anhydrous CH2CI2, and to said solution TEA (3.3 mmoles, 0.46 ml) is added and it is brought to 0°C. l-bromo-3-chloropropane (3.3 moles, 0.32 ml) is cold added, it is brought under reflux (50°C) . After 3 hours the same amount of TEA and of l-bromo-3- chloropropane is added and the reaction is maintained under reflux for 24 hours.
• Prednisolone-21- [2- [4- (3-chloropropyl) piperazin-1-yl] acetate] (0.55 mmoles, 310 mg) is dissolved in 8 ml of anhydrous CH3CN and 6 mi of anhydrous THF, and to said solution AgN03 (1.65 mmoles, 280 mg) ' is added and it is brought under reflux (100°C) under nitrogen, sheltered from the light for 5 hours. It is filtered and the solvent is evaporated at reduced pressure. The residue is purified by chromatography on silica gel using an eluent mixture of AcOEt/MeOH 9:1 (v/v). 155 mg of product have been obtained as a brown solid (48% yield). M.p.: 116 0 -118°C.'
• Example 8 The compound isolated at the end of Example 8 (50 mg) is dissolved in 6 ml of a mixture MeOH/DCM (dichloromethane) (1:1). To the solution cooled at 0°C some drops of a HCl/MeOH solution are added. After 5 minutes at 0°C the solvent is removed at reduced pressure and the residue is treated with ethyl ether. A white solid is formed which is filtered. M.p.: >240°C. Elemental analysis:
• the compound isolated at the end of the previous step (570 mg, 1.1 mmoles) is dissolved in 10 ml of a mixture aceto- nitrile/THF (4:1 v/v) and to the solution, cooled at 0°C, TEA (0.3 ml, 2.15 mmoles) and p-chloro-methylbenzoyl chloride (233 mg, 1.18 mmoles) are added.
• the reaction mixture is brought to room temperature, it is dried after 3 hours, the residue is treated with water (5 ml) and DCM (3x10 ml) .
• the joined organic extracts are washed with brine (5 ml), anhydrified by a2 ⁇ 4 and dried. From the raw product purified by flash- chromatography (DCM/MeOH 9.5/0.5) 731 mg of product (80% yield) are recovered as a white solid. M.p.: 215°-217°C.
• Prednisolone-21- [2- [4- (4' -chloromethylbenzoyloxy) propyl piperazin-1-yl] acetate] (0,82 mmoles, 560 mg) is dissolved in a mixture formed by anhydrous CH3CN (16 ml) and anhydrous THF (12 ml). AgN03 (24.6 mmoles, 418 mg) is added. The mixture is heated under reflux shletered from the light for 3 hours. It is filtered and the solvent is removed at reduced pressure. The residue is purified by chromatography on silica gel, using a mixture AcOEt/MeOH 9/1 (v/v) . 560 mg of product (96% yield) have been obtained.
• Prednisolone-21- [2- [4- (4' -nitrooxymethylbenzoyloxy) propyl piperazin-1-yl] acetate] 50 mg are dissolved in 6 ml of a mixture MeOH/DCM (dichloromethane) (1:1) and to the solution cooled at 0°C some drops of a HCl/MeOH solution are added. After 5 minutes the formed precipitate is filtered obtaining a white solid. Elemental analysis:
• Example 8 The compound isolated at the end of Example 8 (50 mg) is dissolved in 5 ml of acetonitrile. To the solution cooled at 0°C some drops of a trifluoroacetic acid solution (0.4 ml) in acetonitrile (4 ml) are added. After 5 minutes at 0°C the solvent is removed at reduced pressure and the residue is treated with ethyl ether. A white solid is formed which is filtered. Elemental analysis:
• the interactioon between the steroid molecules with specific receptor proteins located in the target organ tissues determines the receptor activation and causes a series of biochemical and physiological transformations inside the tissues, which are the steroid pharmacological effect.
• nitrooxyderivative efficacy according to the present invention and the corresponding nitrooxyderivatives having an aliphatic linking group has been determined in a binding model to a glucocorticoid receptor.
• the corticosteroid binding itself to the receptor activates the human membrane protein CD163, isolated and characterized by Morganelli P.M. et al., J. Immunol., 1988, 140, 2296-2304.
• the isolated cells have been transferred in test tubes (1x10 s cells, the measurements have been carried out in duplicate) containing the culture medium RPMI 1640 and glutamine 1%.
• test tubes (1x10 s cells, the measurements have been carried out in duplicate) containing the culture medium RPMI 1640 and glutamine 1%.
• 3 H-dexamethasone 50 nM in DMSO
• the tested compounds dissolved in the same solvent (DMSO) at the concentrations indicated in the Tables reported hereunder have then been added.
• the test tube content has been mixed using a Vortex equipment.
• the test tubes have then been incubated at 37 °C for 1 hour.
• PBS phosphate buffer cooled in ice bath
• Hydrocortisone-21- (4-nitrooxybutyrate) (Hydr-C 4 -ON0 2 ) prepared as described in patent application WO 98/15568;
• Sprague Dawley normotensive male rats have been divided in groups and treated, respectively, with Prednisolone 21- [ (4 '-nitrooxymethyl) benzoate] (Ex. 3) 5 mg/Kg/die i.p. for 3 weeks and with the corresponding precursor at the same dose.
• the controls have been treated with the carrier (peanut oil 0.5 ml/rat/die i.p. for 3).
• the average arterial pressure (MABP) and the heart-beat have been controlled in the rats.
• NO-Budesonide (635 ⁇ g/ml) and Budesonide (448 ⁇ g/ml) dissolved in a mixture (v/v) DMSO 20%, ethanol 10%, saline physiological solution 70%, or the carrier, have been administered to the animals as aerosols, in a sealed room, using a wright nebulizer operating by compressed air at a pressure of 21.38xl0 3 Pa (20 p.s.i.) and a flow of 0.5 ml/min. The administration lasted 15 minutes.
• prednisolone-21- (4' -nitrooxymethyl) - benzoate vs. prednisolone has been determined in a model of arthritis in rats.
• Lewis female rats weighing 150-200 g fed by a standard diet and with free access to water have been stabulated with cycles of 12 hours light/dark.
• the rats were anaesthetized with halothane (day zero) , then at the base of the tail, by intradermal injection, a collagen suspension II/Freund's incomplete adjuvant (400 ⁇ g/rat) was injected, prepared as described hereinafter: nasal bovine collagen of type II (Sigma- Aldrich, 4 mg/ml) has been dissolved in acetic acid (0.01 M) and emulsified with a same volume of cold Freund' s incomplete adjuvant (Sigma-Aldrich) .
• the rats From the 12th to the 18th day subsequent to the injection, the rats, divided in 3 groups of 10 animals each, have been treated i.p. according to the following protocol: Group 1: prednisolone-21- (4' -nitrooxymethyl) benzoate (4 ⁇ moles/kg) ; Group 2: prednisolone (4 ⁇ moles/kg);
• Group 3 carrier control (peanut seed oil 0.5 ml/kg, i.p. ) .
• a fourth group of healthy rats has been taken as a further reference.
• the anti- arthritic activity has ben evaluated by the following parameters : average paw volume determined by a plethysmometer; clinical evaluation of the hip functionality by an arbitrary score from 0 (absence of inflammation) to 3 (serious inflammation, which affects both the hip articulation and the animal paw) .
• apatite calcium phosphate
• the obtained samples have been analyzed with an inverse microscope (Diaphot TMD; Nikon, Japan) connected to an imagine acquisition system (Argus-10, Hamamatsu Photonics, Enfield, UK) .
• an inverse microscope Diaphot TMD; Nikon, Japan
• an imagine acquisition system Arx-10, Hamamatsu Photonics, Enfield, UK
• the celiac arteries of anaesthetized rats (6 animals/group) have been temporarily occluded with surgical forceps and a HCI solution (1 ml, 0.1 N) has been introduced in O 03/064443 the gastric lumen. After 30 minutes from the introduction of the acid solution the circulation has been reactivated and after 60 minutes from the restarting of the blood circulation the gastric damage has been determined by a lesion intensity index score (LI) .
• LI lesion intensity index score
• Prednisolone-21- (4' -nitrooxymethyl) benzoate and prednisolone (28 ⁇ moles/kg) have been administered to rats by os 2 hours before the ischaemia.
• Rats (no. 3 groups of 10 animals each) have been subjected to a trauma of the spinal cord at the thoracic level by a weight fall (10 g) . In this way a spinal lesion is provoked, which determines a remarkable compromising condition of the motor function.
• rats are treated once a day for 5 days with Prednislone-21- (4' -nitrooxymethyl) benzoate (dissolved in saline solution/ethanol 1:8, 20 mg/kg, s.c.) and Prednisolone (20 mg/kg, s.c, likewise dissolved) or with the only carrier.
• the animal behaviour is evaluated on the third, fifth and seventh day subsequent to the trauma by a multiple score (BBB score) .
• BBB score multiple score
• the zero value is assigned when the condition of the motor function is severely compromised (the animal does not walk) ; the 20 value corresponds to the normal motor functionality.

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