BRPI1100318A2 - A process for obtaining compounds derived from steroidal anti-inflammatory drugs (s), pharmaceutical compositions containing such compounds and their uses in the treatment of inflammatory diseases. - Google Patents

Abstract

PROCEDURE FOR OBTAINING STERAID ANTI-INFLAMMATORY DERIVATIVE COMPOUNDS (EIAs), PHARMACEUTICAL COMPOSITIONS CONTAINING SUCH COMPOUNDS AND THEIR USE IN THE TREATMENT OF INFLAMMATORY DISEASES. The present invention relates to the process of obtaining compounds derived from steroidal antiinflammatory drugs (EIAs) and thalidomide with antiinflammatory, analgesic and immunosuppressive properties useful in the treatment of inflammatory processes. The present invention also relates to pharmaceutical compositions containing such compounds and their uses in the manufacture of medicaments for the treatment of inflammatory diseases, especially those related to chronic inflammatory processes.

Description

Process of Obtaining Steroidal Anti-Inflammatory Derived Compounds (IIAs), Pharmaceutical Compositions Containing Such Compounds and Their Uses for the Treatment of Inflammatory Diseases

The present invention relates to the process of obtaining compounds derived from steroidal anti-inflammatory drugs (EIAs) and thalidomide with anti-inflammatory, analgesic and immunosuppressive properties useful in the treatment of inflammatory processes. The present invention also relates to pharmaceutical compositions containing such compounds and their uses in the manufacture of medicaments for the treatment of inflammatory diseases, particularly those related to chronic inflammatory processes. Fundamentals of the invention

Inflation is fundamentally an answer of

--protection — of— oxga η ± smo__d.e.s_anc postponed by too many physical styles,

----- chemical and biological substances that can cause pain, edema and, in some cases, lead to organ or tissue dysfunction. The inflammatory process is divided into acute and chronic patterns. Acute inflammation is characterized by vascular and exudative phenomena and has a short duration of hours or days. Generally, the acute inflammatory process is treated with non-steroidal anti-inflammatory drugs (NSAIDs). Failure to resolve the inflammation may lead to a chronic condition. Chronic inflammation is characterized by proliferative phenomena, with formation of fibrosis and lasting weeks, months or years.

Chronic inflammation occurs in situations of persistent infection, prolonged exposure to certain

20

25

30 irritants and autoimmune reactions.

Morphologically, the process is characterized by inflammatory infiltrates (edema) generated by mononuclear cells (macrophages, lymphocytes and plasmocytes);

tissue destruction produced by persistence of the offending agent or inflammatory cells; and attempted repair that produces connective tissue. The main cells present in the chronic inflammatory focus are macrophages that are responsible for the production of a series of chemotactic and inflammatory mediators, besides cytokines.

In chronic inflammation, the participation of proinflammatory cytokines plays an important role in the spread and. maintenance of the inflammatory condition. Among the proinflammatory cytokines produced by activated macrophages,

highlight α-interleukin-β (IL-β), IL-6, ~ TL-8 and -o · -a.cro.s.e_tumoxa factor] _alfa _ (_ TNE = cO _._

Tumor necrosis factor alpha. (TNF-a) _ is

biosynthesized by monocytes, activated macrophages, T cells, natural killer cells (NK).

TNF-α enhances chemotactic properties and is responsible for increasing neutrophil adhesion to the vascular endothelium due to stimulation for adhesion molecule production. It stimulates the production of free radicals and the synthesis of other inflammatory mediators such as IL-I and PGE2-0. TNF-α also induces changes in coagulant and anticoagulant properties and increases hepatic synthesis of some acute phase reagents. In addition, TNF-α is an important mediator of septic syndrome and endotoxic shock and is capable of suppressing biosynthesis of lipoprotein lipases and lipogenic enzymes in adipose tissue, impairing lipid storage in adipocytes. The ability of TNF-α to alter vascular endothelial anticoagulant properties and induce procoagulant activity on the endothelial cell surface, stimulating platelet activation factor (PAF) production, and increasing leukocyte adhesion to vascular endothelial cells , results in increased resistance to blood flow, hindering circulation and thus exacerbating microvascular stasis.

TNF-α plays an important role in acute and chronic inflammatory pain. In acute pain, TNF-α sensitizes

neurons ^ nocicept i ν s i_nd_iret ame η t and via_induction of other

proinflammatory cytokines such as IL-β, IL-6, IL-8, resulting in the release of prostaglandins. In chronic "" inflammatory pain, it appears to occur the participation of the "reeep-fe" - ^ - T-N-F = -R1-or-mE ^ p-5-5 -._; _ L______

There are also reports of the importance of TNF-α in peripheral hyperalgesia, and demonstrations that its inhibition is associated with pain reduction. TNF-α also appears to be involved in neuropathic pain, where increased concentration of this cytokine is related to worsening of painful conditions.

Inhibition of this cytokine has been used as a strategy in several diseases such as rheumatoid arthritis, lupus, psoriasis, inflammatory bowel diseases such as Chron's disease, ulcerative colitis, inflammatory eye diseases such as uveitis, asthma, cancer, leprosy and others. Blockage may occur at different levels and may be performed by macro or micromolecules. Among the inhibitors available in therapy are: monoclonal antibodies

(infliximab, etanercept, adalimunab) and micromolecules, represented by drugs structurally related to thalidomide, such as lenalidomide.

Thalidomide, used as a hypnotic and sedative in the 1950s and indicated for the treatment of morning sickness in pregnant women, was banned from the market in 1960 for inducing teratogenicity. However, this drug was reborn in 1998 with FDA approval for use in some diseases such as multiple myeloma, myelodysplastic syndrome, leprosy. These new indications occurred due to the discovery that. Thalidomide could inhibit TNF-α promoting benefits in diseases in which there is an exacerbated increase in this proinflammatory cytokine.

Studies of the "structure-relationship versus activity of

-mo-l-é cu-l-a-de-ta-l-idom ida_t-have shown the relevance of

phthalimide subunit — taken as pharmacophoric. in inhibiting TNF-α. These studies also showed the irrelevance of the glutamiridic subunit in this action. From this knowledge, and using the tools of pharmaceutical and medicinal chemistry, it is possible, keeping the primary pharmacophoric group, to plan bioactive structures in the inhibition of TNF-α devoid of toxicophoric groups, responsible for the teratogenic effect.

From this line of reasoning, it is desirable to plan and study new thalidomide analogs since they more effectively and safely inhibit TNF-α and thus constitute an alternative to the treatment of chronic inflammatory processes. The treatment of inflammatory processes occurs mainly with the use of anti-inflammatory drugs that may or may not be steroidal in nature. These are able to interfere with the body's reaction process of defense in order to minimize damage.

Steroidal anti-inflammatory drugs (EIAs), known as glucocorticoids (GC), have been widely used for their potent anti-inflammatory and immunosuppressive activity since the 1950s. Among the EIAs, hydrocortisone, dexamethasone, triamcinolone, cortisone, prednisone have been highlighted. fludrocortisone betamethasone methyl

prednisolone, prednisolone, flurandrenolide, fluocinolone,

beclomethasone, budenoside, flunisolide, triamcinolone acetonide. These drugs act by inhibiting the production of

prosLaglandins and leukotrienes by the —inhibitory action of -of-s-f-θ-1-i-if-A-2 -, - by-1-ib-exaction_of_mediator_pr_o_tAi.c_o_

-—- anti-inflammatory liporcortin-1. GCs affect all types of inflammatory reactions from reactions caused by invasive pathogens to inadequately developed immune responses such as those observed in hypersensitivity reactions or autoimmune disease. They are also used clinically to suppress graft rejection by suppressing the triggering and production of a new immune response more effective than the one already triggered. They act on cellular targets that regulate gene expression and protein biosynthesis via formation of steroid receptor complexes. Thus, they may reduce the transcription of various inflammatory enzymes / proteins such as

cyclooxygenase-2 (COX-2), induced nitric oxide synthase (iNOS) and some proinflammatory cytokines.

The different GCs can be administered orally, intravenously, intramuscularly, intranasally or topically and differ in pharmacokinetic and pharmacodynamic profile, as well as in their glucocorticoid actions rather than mineralocorticoids. Among the actions of glucocorticoids (GCs), the following stand out:

a) lower blood vessel neutrophil output and reduced neutrophil and macrophage activity due to decreased transcription of cell adhesion factor genes and cytokines;

b) decreased T-helper action and reduced β-clonal proliferation of Τ cells, primarily through decreased transcription of IL-2 genes;

c) decreased fibroblast function and, therefore, lower production of collagen and glyososaminoglycans;

-d.) - r-education_of_the_function_of_the_ steoblastas_e_maior

osteoclast activity- - --____________.

e) decreased production of prostanoids due to reduced expression of COX-2;

f) reduction in cytokine production: IL-1, IL-2, IL-3, IL-4, IL-5, IL-6, IL-8 and cell adhesion factors;

g) reduction in plasma concentration of complement components;

h) reduction in induced nitric oxide production;

i) lower histamine release from basophils; and

j) decreased IgG production.

Glucocorticoids are still used for the treatment of asthma and allergic diseases, usually in combination with other drugs. Moreover, they have important immunosuppressive applications in organ transplantation. Under organ transplantation conditions, corticosteroids are used in combination with other drugs such as calcineurin inhibitors (cyclosporine or tacrolimus) and antiproliferative agents.

Over the years, the esterified spheroidal anti-inflammatory derivatives available in therapy have been modified to obtain a more appropriate pharmacokinetic profile, ie:

a) more fat-soluble forms that have greater absorption and a slow (prolonged) release profile - generally used in the intramuscular route of administration;

(b) more hydrophobic forms for the use of intravenous or intramuscular routes, especially in emergency conditions that should be treated with steroidal anti-inflammatory drugs. '' ......

-A-p-Eednisone-and-a-p-redn-i-so-Canvas-are-glucocorticoids

(GCs) of - action ...... intermediate and were introduced - in the

therapy one year after fludrocortisone. These were more active as antirheumatic and antiallergenic agents and produced fewer undesirable adverse effects. These drugs have about 80% availability, high percentage protein binding (90%), and plasma half-life ranging from 2-4 hours. In addition, they present less mineralocorticoid action than hydrocortisone.

About 20 prednisolone ester derivatives are available, the fundamental difference of which is centered on the pharmacokinetic profile and / or pharmacotechnics (with modifications of administration routes). Prednisolone butyl acetate ester, available for injection use only, has a long duration of action due to its low water solubility and slow hydrolysis rate. Prednisolone sodium phosphate is a water-soluble sodium salt with a half-life of less than 5 minutes due to rapid hydrolysis by phosphatases.

These strategies used to modulate the pharmacokinetic and pharmacotechnical profile of steroidal anti-inflammatory drugs are called molecular modification.

Molecular modification is one of the strategies used for the discovery of new drugs. It basically consists of, from the prototype of

known biological structure and structure. _ ^ synthesize

counterparts, analogues or structural homologues thereof in order to alter their biopharmaceutical properties. Biopharmaceutical "properties" means the properties of

—F-is-i-Co = chem-cas_or_pharmacies_and_as_b-i-olo-g-___ Atraxés

of molecular modification one can discover ____ the ______ group (s)

pharmacoform (s), that is, the subunit (s) of the drug molecule responsible for its activity - and thus obtain better structures than those from which they derive, either in potency, toxicity and specificity, in duration, stability and even in relation to the cost of production. The goal of molecular modification is to perform chemical transformations of known active ingredients to obtain compounds with higher potency, better specific activity profile, better safety, better formulation for handling by health professionals, and better patient acceptability.

Latency is the transformation of the drug into an inactive transport form, which in vivo, by chemical or enzymatic reaction, releases the active subunit at or near the site of action. Latency is an important strategy for drug insertion in therapy.

The compounds obtained by this strategy are called prodrugs. Among the prodrugs available in therapy are enalapril, omeprazole, simvastatin and acyclovir, considered blockbusters due to their market potential.

Prodrugs are designed to overcome pharmaceutical and pharmacokinetic problems associated with the matrix molecule, which otherwise could have limited clinical use. Urruugrande. ^ Number ..of_J? A.rreIras _may ^^ drug.

In the pharmaceutical phase, related to the obtainment of the method, 'podo-s and "—o b s crvar; d e η t rc' or t r s sis as -; - f a It a

de-s-o-l-ub-i-l-id-ade-T-e-s-fea-b-i-1-id of -, - and -property

undesirable organoleptic agents. Solubility is often altered by modulating carrier activity by the introduction of fat-soluble or water-soluble groups. This type of change does not alter intrinsic activity of the drug after carrier release in vivo. Stability, likewise, can be modulated by electronic and steric effects, assisting in the stages of pharmacotechnical development. The properties

Organoleptic compounds can also be altered by synthesizing the prodrug which often has the ability to mask unpleasant tastes.

It is in the pharmacokinetic phase that the use of latency has been used with most interest. In this phase, the absorption, distribution, metabolism, excretion and toxicity of the drugs are taken into consideration. Thus, later stages of drug development have been a barrier to a number of ligands identified by the molecular modeling strategy. Many of these pharmacokinetic problems can be solved by the latency technique.

When designing a prodrug, consideration should be given to: the existence of functional groups in the matrix molecule capable of derivatization; the existence of mechanisms and / or systems in the body capable of bioactivating the prodrug; the ease and simplicity of

The purification of the prodrug; a_____stability

prodrug chemistry; the in vivo regeneration of the matrix molecule in optimal amounts; and the toxicity of the carrier and "prodrug" per se. - ■ -

-Following-i-me-n-t-Q -, - are-cited-some-do.c.umen.tas

related to this technology. However, everyone has differences in the process and compounds which are objects of this invention.

Patent application PI 0316240,. filed on 11/13/2003, corresponding to international publication WO 2004/045579, describes pharmaceutical compositions and unit dosage forms of thalidomide and their pharmaceutically acceptable salts, solvates, hydrates or clathrates. Such patent application also refers to methods of treating and preventing leprosy, chronic graft-versus-host disease, rheumatoid arthritis, sarcoidosis,

inflammatory bowel disease and cancer by employing new dosage forms. However, in this document there is no change in the chemical structure of thalidomide. This is at most derivatized into respective salts to alter solubility patterns.

International publication WO 97/37988 of 10/16/1997 describes some thalidomide analogues that inhibit angiogenesis in vivo in humans and animals. In WO 97/37988, inactive carriers have been inserted into thalidomide derivatives such as alkyl, urea, guanidinic and amino acid groups in order to alter only pharmacokinetic parameters. The resulting compounds in their cited publication are unrelated to the compounds described in this invention. Accordingly, thalidomide derivatives described in WO 97/3798888 do not. show action synergism, previously thought to be beneficial in the compounds now invented and described, since we observe "that the introduction of an anti-inflammatory

you-pooh-po-tency-1-i-za-the-anti-action. mud. Toxia_e_

immunosuppressive --------------------------. -, -__________________

International publication WO 97/45117 'of 14/12/1997 describes thalidomide prodrugs with differentiated physicochemical characteristics, such as better solubility. Prodrugs are proline-terminated dipeptide structures that can be characterized as follows: thalidomide-X-proline, where X may be different amino acids that are considered inactive transporters. The compounds of this document bear no resemblance to those proposed in the present invention.

The present invention, using the latency strategy, proposes the synthesis of steroidal anti-inflammatory derivatives (EIAs), more specifically prednisolone derivatives, and thalidomide derivatives to obtain prodrugs. The main advantages of the technology developed in the invention are pharmacokinetic and pharmacodynamic. From the pharmacodynamic point of view, the "association" of two subunits with anti-inflammatory and analgesic properties provokes a synergism of action, in order to increase the potency of these activities. The compounds obtained in the present invention act by synergism of action in the inhibition / modulation of inflammatory mediators, mainly tumor necrosis factor alpha (TNF-α). In addition, these compounds still have immunomodulatory properties, allowing

its use as an immunosuppressant in processes for____

organ rejection. From a pharmacokinetic point of view, the more lipophilic character allows, as previously adopted

for —IE esters, slow-release systems --- and-

s_t_files_entry_entry, _reducing_a.s_S-im, _a._neces.aity_of

multiple administrations, facilitating use and improving patient compliance with therapy.

Brief Description of the Invention

The present invention aims to synthesize and obtain prodrugs derived from steroidal anti-inflammatory drugs and thalidomide derivatives with potential anti-inflammatory effects due to inhibition / modulation of proinflammatory cytokines such as tumor necrosis factor alpha (TNF). -α). The compounds (prodrugs) obtained have anti-inflammatory activity and are potentially useful in the treatment of chronic inflammatory diseases. The present invention also relates to pharmaceutical compositions and uses containing such compounds. Brief Description of the Figures

Figure 1 shows the general structural planning and formation of new compounds from steroidal anti-inflammatory drugs and thalidomide.

Figure 2 shows carrageenan-induced rat paw edema.

Figure '3 shows carrageenan-induced rat paw edema in the presence of prednisolone (standard) and prednisolone 5 obtained in example 1.

Detailed Description of the Invention

This invention features as an innovative feature the synthesis of novel anti-inflammatory derivatives designed

a_party .. _anti-iηf 1 amajor _ steroidsu ^ protos;

glucocorticoids (1) and thalidomide (2), rationally designed through the molecular hybridization strategy as "phospholipase A2 inhibitors" and modulators of TNF-α with activity-an-td— in-f-mud-to-ti-a — and / or — analgesic (Figure 1). -., ... ,, - ■. ............-

The new derivatives were obtained in good to excellent chemical yields, using synthetic methodology as described in the scheme of figure 1, which has few synthetic steps, starting from commercially available compounds, which qualifies this synthetic methodology to industrial use.

The compounds of the present invention were designed through divergent syntheses using classical reactions such as:

• Regioselective Nucleophilic Substitution;

• Nucleophilic Addition to Carbonyl; • Condensation for imida formation. Specifically, the process comprises the steps of: Preparing suitably functionalized thalidomide derivatives by condensing suitably functionalized amines to phthalic anhydride using acetic acid or pyridine as solvents. Obtaining these compounds can also be obtained using domestic microwave radiation at times shorter than those obtained by conventional heating.

Nucleophilic substitution to obtain esters

using coupling agents (such as dicyclohexylcarbodiimide - DCC) in inert solvents (such as diclojromethane, chloroform, pyridine) at low.

temperatures at varying times. More specifically, the glucocorticoid derivatives (C) of the present invention may be prepared according to the synthetic route shown below.

20

Wf

η η H0?

VfT ♦

The

glucocorticoid thalidomide derivative (B)

(THE)

i) coupling agent, inert solvent, O0Cl 0.5-6 h (35-80%)

Thalidomide derivatives (A) are compounds which

present the structural formula shown in the above synthetic route:

Where:

R = C 6 H 4 (phenyl), (CH 2) n where η = 1, 2, 3, 4, 5, 6, 7, 8 or 9, C 5 H 3 N, C 4 H 3 N 2, C 3 H 2 N 3, NH, 2-hydroxyphenyl, 3-hydroxy -phenyl, 4-hydroxy-phenyl, 2-hydroxy-benzyl, 3-hydroxy-benzyl, 4-hydroxy-benzyl, 2-hydroxy-ethylbenzyl,

3-hydroxyethylbenzyl, 4-hydroxyethylbenzyl, benzyl, thiophene, furan, pyrrole, 2-hydroxypyridine, 3-hydroxypyridine, 4-hydroxypyridine, pyrazine, pyrimidine, benzothiophene, benzofuran, indole, quinoline, isoquinoline, naphthalene, CH2-2-hydroxythiophene, CH2-3-hydroxythiophene, CH2-2-hydroxy-furan, CH2-3-hydroxy-furan, CH2CH2-2-hydroxy-thiophene, CH2CH2-3-hydroxy- thiophene, CH 2 CH 2 -2-hydroxy furan, CH 2 CH 2 -3-hydroxy furan, glutarimide, hydroxymethyl glutarimide, succinylhydroxymethylglutarimide, phthalylhydroxymethylphthalimide, 2-aminophenyl, 4-aminophenyl, 2-amino-benzyl , 3-amino benzyl,

4-amino-benzyl, 2-amino-ethylbenzyl, 3-amino-ethylbenzyl, 4-amino-ethylbenzyl, 2-amino-pyridine, 3-amino-pyridine, 4-amino-pyridine, CH2-2-amino-thiophene, CH2-3-aminothiophene,

CH2 ^ 2 ^ amino ^ furan _C2 H2--. 3 - a myofuran, _CH2-CH2-2-amino-

thiophene, CH 2 CH 2 r-3-amino thiophene. CH2CH2-2-amino furan .... or. CH 2 CH 2 -3-amino-furan, 3-methyl-2,6-dioxo-piperidin-1-yl-methyl-propyl ester;

W = H, (4 and / or 5 and / or 6 and / or 7) -F; (4 and / or 5 and / or 6 and / or 7) -Cl; (4 and / or 5 and / or 6 and / or 7) -Br; (4 and / or 5 and / or 6 and / or 7) -NO 2; (4 and / or 5 and / or 6 and / or 7) -NH 2; (4 and / or 5 and / or 6 and / or 7) -OH; (4 and / or 5 and / or 6 and / or 7) -OCH 3; (4 and / or 5 and / or 6 and / or 7) -OCF3; (4 and / or 5 and / or 6 and / or 7) -CF 3; and

The selected glucocorticoids (B) are:

hydrocortisone, dexamethasone, triamcinolone, cortisone, prednisone, fludrocortisone, betamethasone,

methylprednisolone, prednisolone, flurandrenolide,

fluokinolone, desonide, alclometasone, flumetasone, deoxymethasone, beclomethasone, budenoside, flunisolide, triamcinolone acetonide;

Limiting glucocorticoid to prednisolone gives the synthetic route used for the synthesis of the compounds of the invention. Such synthesis route is shown below:

(THE)

The following describes in detail the synthesis of the following compounds:

Example 1 - (8S, 9S, 10R, JL3S, 14S, 17 ') 7,8,9,11,12,13,15,15,16,17-decahydro-11,17-dihydroxy-17- (2 - (1,3-dioxoisoindolin-2-yl) acetyl acetate-10,13-dimethyl-6H-cyclopenta [a] phenanthrene-3 (10H) -one (prednisolone prodrug-5) -;

Example 2 - (8S # 9S, 10R, 13S. 14S. 17R) -7. 8 9 11. 12. 13L.

14,15,16,17-dehydro-11,17,77, dihydroxy-17- (4- (1,3-dioxoisoindol-2-yl) benzoate acetyl -10,13-dimethyl-6H-cyclopenta [a] phenanthrene-3 (IOH) -one (prednisolone prodrug-8);

Example 3 - (8S, 9S, 10R, 13S, 14S, 17R) -7,8,9,11,12,13,11,15,16,17-decahydro-11,17-dihydroxy-17- (3- (1,3-dioxoisoindolin-2-yl) benzoate acetyl-10,13-dimethyl-6H-cyclopenta [a] phenanthrene-3 (10H) -one (prednisolone prodrug-11);

Example 4 - 8S, 9S, 10R, 13S, 14S, 17R) -7,8,9,11,12,13,11,15,16,17-decahydro-11,17-dihydroxy-17- (2- ( 1- (hydroxymethyl succinyl) -2,6-dioxopiperidin-3-yl) isoindoline-1,3-dione acetyl-10,13-dimethyl-6H-cyclopenta [a] phenanthrene-3 (10H) -one (prodrug prednisolone - 15);

EXAMPLES Example 1) Synthesis of prednisolone-derived prodrug (8S, 9S, 10R, 13S, 14S, 17R) -7,8,9,11,12,13,14,15,16,17-decahydro-11, 17-dihydroxy-17- (2- (1,3-dioxoisoindolin-2-yl) acetyl acetate-10,13-dimethyl-6H-cyclopenta [a]

phenanthrene-3 (IOH) -one (5)

a) Preparation of 2- (1,3-dioxoisoindolin-2-yl) acetic acid phthalimide derivative (3)

The 2- (1,3-dioxoisoindolin-2-yl) acetylic acid phthalimide derivative (3) 'was synthesized according to the methodology previously described in the literature. Reaction

_0.05 mol of ani.dr_ldo_f_t al.i_c.o __ (- 1 -) -, - 0 -, _ 0-5 — mo-1 — de-güe-i-na

(2), 30 mL of toluene - and 0.5 mL of triotane.lamine were added in a 125 mL flask, connected to a refluxed dean stark. The reaction mixture was kept under vigorous stirring and heating for 4 hours. After cooling the reaction mixture, the organic solvent was evaporated under reduced pressure and at the end of this process a white solid was obtained. 75 mL of ethyl acetate was added to this white solid and with the aid of a separatory funnel, the organic phase was washed 4 times with 40 mL of 1M HCl solution. After isolation, anhydrous sodium sulfate was added, filtered and the organic phase was evaporated under reduced pressure yielding 1.95g of a white powder (3) whose molecular formula is C10H11O4N, MW = 205.17, RF = O , 84 and MP = 187-193 ° C. The reaction yield was 79.5%.

1 H NMR (300 MHz; CDCl 3 d): δ 7.81-7.90 (m, 4H); 4.4 (s; 2H) ppm.

C13 NMR (300MHz; CDCl3d): δ 170; 168.3; 134.7; 131.1;

124.2; 39.1 ppm.

Infrared (KBr pellet): υ Aromatic C-H = 3084 cm -1; υ alkyl C-H (symmetrical and asymmetrical) = 2927 and 2850 cm -1; υ C = O imide (symmetrical and asymmetrical) = 1786 cm-1 and 1717 cm "1; υ C = Acid = 1697 cm-1; υ C = Aromatic C = 1560 cm-1 and 1450 cm-1; υ CNC 1381 cm -1.

(b) Preparation of the prednisolone prodrug (5): (8S, 9S, 10R, 13S, 14S, 17 R1 - IQ, 9, 12, IJ, 14, 1,5,1617-dec. idro-11). .17-dihydroxy-17- (2- (1,3-dioxoisoindolin-2-yl) acetyl acetate-10,13-dimethyl-6H-cyclopenta [a] phenanthrene-3 (10H) -one. Reaction-

170 mg (0.83 mmol) of 2- (1,3-dioxoisoindolin-2-yl) acetic acid phthalimide derivative (3) was dissolved in mL of anhydrous dichloromethane in a 25 mL flask connected to a tube with drying, forming a solution that was kept at 0 ° C. 172 mg (0.83 mmol) dicyclohexylcarbodiimide (DCC) and 11 mg (0.83 mmol) dimethylaminopyridine (DMAP) were added to said solution. The mixture was kept under stirring at 0 ° C for 15 to 20 minutes. After this time, 3 drops of triethylamine and 300 mg (0.83 mmol) of prednisolone (4) were added to the solution. The mixture was stirred for 3 hours and followed by thin layer chromatography until completion of the reaction, where the mobile phase was a mixture of ethyl acetate and hexane in a ratio of 8: 2 and the stationary phase was composed of silica.

The reaction mixture was filtered and the filtrate was diluted with about 50 mL of dichloromethane. In a separatory funnel, the organic phase was washed 3 times with about 15 mL of 0.02M bicarbonate solution. After this step, anhydrous sodium sulfate was added to the organic phase. The product was filtered off and evaporated by reduced chromatographic column purified reaction yielding 215 mg of a white solid (prednisolone prodrug (5)) whose molecular formula is C31H33NO8; Mp = 54-7.22; mp 201-204 ° C. "Reaction yield was 47% .---------

1 H NMR (300 MHz; CDCl 3 d): δ. 7.81-7.90 (m; 4H); 7.32 (d; 1H;); 6.14 (d; 1H); 5.91 (s; 1H); 5.2 (s; 2H); 4.94 (s; 2H); 3.3 (s; 1H); 2.56 (d; 2H); 1.8-2.1 (m; 7H); 1.43-1.49 (m; 4H); 1.36 (s; 3H); 0.9 (s; 3H) ppm.

C13 NMR (300 MHz; CDCl3d): δ 211.1; 183.2; 170; 168.3 157.1; 134.9; 130.5; 128; 124.2; 120.1; 89.8; 69.2; 65.3 59.8; 50; 4 5.4; 40; 39.7; 37; 33.3; 32.1; 30.2; 29.7; 22 19.8; 17.1 ppm.

Infrared (KBr pellet): υ O-H = 3327 cm -1;-C-Haromatic = 3084 cm -1; υ C-Halkylic (symmetrical and asymmetrical) = 2943 and 2850 cm "1; υ C = Oimide (symmetrical and asymmetrical) = 1761 cm" 1 and 1705 cm "1; υ C = Oester = 172 9 cm" 1; C = Caromatic = 15 61 cm -1 and 1452 cm -1; υ CNC = 1380 cm "1. Example 2) Synthesis of prednisolone-derived prodrug (8S, 9S, 10R, 13S, 14S, 17R) -7,8,9,11,12,13, 14, 15 , 16,17-decahydro-11,17-dihydroxy-17- (4- (1,3-dioxoisoindolin-2-yl) benzoate acetyl -10,13-dimethyl-6H-cyclopenta [a] phenanthrene-3 (IOH) -ona (8)

(a) The preparation of 4- (1,3-dioxoisoindolin-2-yl) benzoic acid phthalimidic derivative (7)

The 4- (1,3-dioxoisoindolin-2-yl) benzoic phthalimidic derivative (7) was synthesized according to the methodology previously described in the literature.

1 g (7.29 mmol) of p-aminobenzoic acid - (- 6 -) -, - 1 — g

(6.75 mmol) phthalic anhydride (1) and 10. mL .- of glacial acetic acid were added in a 25 mL flask. The reaction mixture was heated at 120 ° C for 1 hour and stirred vigorously. After cooling, the reaction mixture was filtered and the white solid formed was washed with about 60 mL of ice water. Then the obtained solid was recrystallized from 95% ethanol yielding 1.62 g of white crystals (7) whose molecular formula is C 14 H 9 NO 4, MW = 267 and MP = 250 ° C. The reaction yield was 91%.

1H-NMR (400 MHz; DMSOd6): δ 7.96 (m; Hx and H2; 4H); 7.68 (dd; Hge H7; Jorto = 8.1Hz; Jmeta = 2.1Hz; 2H); 7.55 (dd; Hi0 and H6; Jorto = 8.1Hz; Jmeta = 2.1Hz; 2H) ppm.

Infrared (KBr tablet): υ O-H = 3327 cm -1;

Reaction

0

0

(1)

(6) 10

Aromatic CH = 3084 cm "1; υ alkyl CH (symmetrical and asymmetric) = 2927 and 2850 cm -1; υ C = O imide (symmetrical and asymmetrical) = 1786 cm -1 and 1717 cm -1; Ό C = O acid = 1715 cm "x; C = N furoxane = 1573 cm -1; C = aromatic C = 1560 cm -1 and 1450 cm -1; C-N-C 1381 cm-1.

(b) Preparation of the prednisolone prodrug (8): (8S, 9S, 10R, 13S, 14S, 17R) -7,8,9,11,12,13,14,15,16,17-decay 11,17-dihydroxy-17- (4- (1,3-dioxoisoindolin-2-yl) benzoate acetyl-10,13-dimethyl-6H-cyclopenta [a] phenanthrene-3 (1OH) -one.

The

The

= No

OH

15

(7)

(4)

Π ° = · -> I to ^ HO ..

H

222 mg (0.83 mmol) of the phthalimid acid derivative — 4 -

(1,3-Dioxoisoindolin-2-yl) benzoic acid (7) were dissolved in mL of anhydrous dichloromethane in a 25 mL flask connected to a drying agent tube to form a solution which was maintained at 0 ° C. 172 mg (0.83 mmol) dicyclohexylcarbodiimide (DCC) and 11 mg (0.83 mmol) dimethylaminopyridine (DMAP) were added to said solution. The mixture was kept under stirring at 0 ° C for 15 to 20 minutes. After this time, 3 drops of triethylamine and 300 mg (0.83 mmol) of prednisolone (4) were added to the solution. The mixture was stirred for 4 hours and followed by thin layer chromatography until completion of the reaction, where the mobile phase was a 6: 4 mixture of ethyl acetate and hexane and the stationary phase was silica. .

The reaction mixture was filtered and the filtrate was diluted with about 50 mL of dichloromethane. In a separatory funnel, the organic phase was washed 3 times with about 15 mL of 0.02M bicarbonate solution. After this step, anhydrous sodium sulfate was added to the organic phase. The product was filtered and evaporated under reduced pressure and purified by column chromatography affording 265 mg of a white solid (prednisolone prodrug (8)) whose molecular formula is C36H35NO8; MW = 609.66; MP: 188 ° -191 ° C. The reaction yield was 52.3%.

1 H NMR (300MHz; CDCl 3 d): δ 7.87-7.96 (m; 4H) _; _7, _57 '^' ^ (ddT "" 2HT; "7H," 4 4 (dd 2H) 6.3 (d; 1H;); 6.14 (d; 1H); 6.01 (s; 1H); 5.54 (s; 2H); 4.94 (s; 2H); 3 1.3 (s; 1H); 2.56 (d; 2H); 1.8-2.45 (m; 711); -1.4 -3_-1.4 (m; -4H); -, 36 - (s; 3H) v. "07" (s; 3H) ppm.

C13 NMR (300MHz; CDCl3d): δ 211.3; 183.1; 1666; L67.3; 157.2; 135.1; 134.7; 131; 130.1; 129.8; 126.7; 128.2; 123.4; 120.3; 89.7; 69.1; 65.4; 59.6; 50.2; 45.4; 39.6; 37.2; 33; 32.2; 30.4; 29.5; 22; 19.6; 17.3 ppm.

Infrared (KBr pellet): υ O-H = 3327 cm -1; υ Aromatic C-H = 3084 cm "1; υ Alkyl C-H (symmetrical and asymmetrical) = 2927 and 2851 cm-1; υ C = O Imida (symmetrical and asymmetrical) = 1772 cm" 1 and 1707 cm-1; υ C = The ester = 1734 cm "

1; υ C = Ocetone = 1707 cm-1; υ C = Aromatic C = 1561 cm-1 and 1452 cm-1; C-N-C 1380 cm -1.

Example 3) Synthesis of prednisolone-derived prodrug (8S, 9S, 10R, 13S, 14S, 17R) -7,8,9,11,12,13,14,15,16,17-decaihydro-11,17 -dihydroxy-17- (3- (1,3-dioxoisoindolin-2-yl) benzoate acetyl-10,13-d-methyl-6H-c-clopenta [a]

phenanthrene-3 (IOH) -one (11)

a) Preparation of 3- (1,3-dioxoisoindolin-2-yl) benzoic acid phthalimidic derivative (10)

The 3- (1,3-dioxoisoindolin-2-yl) benzoic phthalimidic derivative (10) was synthesized according to the methodology previously described in the literature.

Reaction

H2N

1st WO

^ = / OH

o) o °)

M-Aminobenzoic acid Ig (7.29 mmol) (9), 1 g (6.75 mmol) phthalic anhydride (1) and 10 mL glacial acetic acid were added in a 25-ml flask. mL. The reaction mixture was heated at 120 ° C for 1 hr.

vigorously. After cooling, the reaction mixture was filtered and the white solid formed was washed with about 60 mL of ice water. Then the obtained solid was recrystallized from 95% ethanol yielding 1.39 g of white crystals (10) whose molecular formula is C 14 H 9 NO 4, MW = 267 and MP = 250 ° C. The reaction yield was 78%.

1H-NMR (400 MHz; DMSOd6): δ 8.05 (H8; JmeCa = 1.5Hz; 1H); 8.01 (H6; Jmeta = 1.5Hz; 1H); 7.99 (m; H2; 2H); 7.92 (m; H1; 2H); 7.72 (H10; Jorto = 7.75 Hz; Jraeta = 1.4 Hz; 1H); 7.66 (Hg, - Jorto = 7.92 Hz; 1H) ppm.

Infrared (KBr pellet): υ O-H = 3327 cm -1; υ Aromatic C-H = 3084 cm "1; υ Alkyl C-H (symmetrical and asymmetrical) = 2927 and 2851 cm-1; υ C = O imide (symmetrical and 10

asymmetric) = 1786 cm -1 and 1717 cm -1; C = O ester = 1734 cm-1 C = Ocetone = 1707 cm-1; C = Aromatic C = 1560 cm-1 and 1450 cm-1; C-N-C 1380. cm -1.

b) Preparation of the prednisolone prodrug (8S, 9S, 10R, 13S, 14S, 17R) -7, 8, 9, 11, 12, 13, 14, 15, 16, 17-decal-11, 17- dihydroxy-17- (3- (1,3-dioxoisoindolin-2-yl) benzoate acetyl-10,13-dimethyl-6H-cyclopenta [a] phenanthrene-3 (1OH) -one (11).

(11)

, (4)

222 mg (0.83 mmol) of the phthalimidic acid derivative 3-

(1,3-dioxoisoindolin-2-yl) benzoic (10) were dissolved

------- on — 5 — m-L · —of — i-will-rome-tano-anhydrous, _in_jam. 25 mL flask,

connected to a pipe with ■ drying agent. forming a solution that was kept at 0 ° C. 2 mg (0.83 mmol) dicyclohexylcarbodiimide (DCC) and 11 mg (0.83 mmol) dimethylaminopyridine (DMAP) were added to said solution. The mixture was kept under stirring at 0 ° C for 15-20 minutes. After this time, 3 to 5 drops of triethylamine and 300 mg (0.83 mmol) of prednisolone (4) were added to the solution. The mixture was kept under stirring for 5 hours and followed by thin layer chromatography until the completion of the reaction, where the mobile phase was a 6: 4 mixture of ethyl acetate and hexane and the stationary phase was. composed of silica. The reaction mixture was filtered and the filtrate was diluted with about 50 mL of dichloromethane. In a separatory funnel, the organic phase was washed 3 times with about 15 mL of 0.02M bicarbonate solution. After this step, anhydrous sodium sulfate was added to the organic phase. The product was filtered and evaporated under reduced pressure and purified by column chromatography affording 243 mg of a white solid (prednisolone prodrug (11)) whose molecular formula is C36H35NO8, MW = 609.66; MP: 146 ° -149 ° C. The reaction yield was 48%.

1 H NMR (300MHz; CDCl 3 d): δ 8.3 (s; 1H); 8.05 (d; 1H); 7.92-7.99 (m; 4H); 7.72 (d; 1H); 7.66 (dd; 1H) 6.3 (d; 1H;); 6.12 (d; 1H); 6.02 (s; 1H); 5.53 (s; 2H); 4.92- (s; _2Η) __; 3 ^ 31 Js; 1H); 2.58 (d; 2H); 1.81-2.19 (m; 7H); 1.43-1.49 (m; 4H); 1.36 (s; 3H); 1.09 ('s7 3h1 "ppmT" "

C13 NMR (300MHz; CDCl3d): δ 211.1; 183.2; 166; 167.3; • 157-. 1-; - 134.4; 132.7; 131.2; .130.8_; _287.7; 126.7; 125.5;

69.2; 65.3; 59.8; 50; 45.4; 39.7; 37; 33.J3; 32.1; 30.2; 29.7; 22; 19.8; 171T ~ ppm ~.

Infrared (KBr pellet): υ O-H = 3327 cm -1; υ Aromatic CH = 3084 cm "1; υ Alkyl CH (symmetrical and asymmetrical) = 2927 and 2851 cm-1; υ C = O imide (symmetrical and asymmetrical) = 1772 cm-1 and 1718 cm-1; υ C = O ester = 1734 cm "υ C = Ocetone = 1707 cm" 1; υ C = Aromatic C = 1561 cm "1 and 1452 cm" 1; υ CNC 138 0 cm "1.

Example 4) Synthesis of prednisolone-derived prodrug 8S, 9S, 10R, 13S, 14S, 17R) -7,8,9,11,12,13,14,15,16,17-decaihydro-11,17- dihydroxy-17- (2- (1- (hydroxymethyl succinyl) -2,6-dioxopiperidin-3-yl) isoindoline-1,3-dione acetyl-10,13-dimethyl-6H-cyclopenta [a] phenanthrene-3 ( 10H) -one (15) (a) Preparation of the thalidomide derivative: [3- (1,3-dihydro-1,3-dioxo-2H-isoindolyl-2-yl) -2,6-dioxo-piperidin-1-one ylmethyl] ester (14)

The thalidomide derivative: 3- (1,3-dihydro-1,3-dioxo-2H-isoindolyl-2-yl) -2,6-dioxo-piperidin-1-yl-methyl] ester (14) was synthesized to from 2- (1- (hydroxymethyl) -2,6-dioxopiperidin-3-yl) isoindoline-1,3-dione (13) as previously described in the literature. Reaction

b) Preparation of 2- (1- (hydroxymethyl) -2,6-dioxo piperidin-3-yl) isoindoline-1,3-dione (13)

„. 1/29 g (.50 mmol) of thalidomide (-12) was suspended in ml of a 35% aqueous formaldehyde solution. The suspension was kept under reflux and constant magnetic stirring until complete dissolution. The reaction medium was cooled and refrigerated for a few days until precipitate formation ceased. The precipitate was collected by filtration and washed with a 3% formaldehyde solution, yielding 10.1 g of a white solid product (13), whose molecular formula is C14Hi2N2O5; MW = 288.26 and MP: 165Â ° C The reaction yield was 70%.

1H-NMR (300 MHz; CDCl3d6): δ 7.92-7.98 (m; 4H); 6.15 (t, Jab = 7.2 Hz, Jbc = 1.2 Hz, 1H, OH); 5.5 (m, 1H, CHCH 2); 5.1 (d, Jab = 7.2 Hz, 2H, NCH 2 OH); 2.88-3.14 (m, 2H, H-5 '); 2.17-2.69 (m, 2H, H-4 ') ppm. "

Infrared (KBr pellet): υ O-H = 3207 cm -1; Aromatic C-H = 3084 cm -1; Alkyl C-H (symmetrical and asymmetric) = 2972 and 2851 cm -1; υ C = O imide (symmetrical and asymmetrical) = 1782 cm -1 and 1711 cm -1; υ C = Aromatic C = 1560 cm -1 and 1454 cm -1; C-N-C 1380 cm -1. (c) Preparation of 3- (1,3-dihydro-1,3-dioxo-2H-

isoindolyl-2-yl) -2,6-dioxo-piperidin-1-yl-methyl] ester (14) 475 mg of 2- (1- (hydroxymethyl) -2,6-dioxopiperidin-3-yl) isoindolin-2-one 1,3-dione (13), 330 mg succinic anhydride, 30 mg dimethylaminopyridine (DMAP), 0.5 mL triethylamine and approximately 8.5 mL distilled dichloromethane were placed in a 125 mL flask connected to a tube containing drying agent. The reaction mixture was added under stirring at room temperature for 24 hours. Then the reaction mixture was extracted 3 times with 50 ml of a 5% hydrochloric acid solution and 3 times with 50 ml of water. .The Phase.

------- was -aeca-.com__sulfat 13 Hydro and the solvent was

evaporated under reduced pressure. The residue was crystallized from ethyl acetate and recrystallized from ethanol to afford a solid product (14) in 68% yield.

1 H NMR (300 MHz; CDCl 3 CF 6): δ 7.86-7.90 (m, 2H); 7.76-7.79 (m, 2Ή), 5.84, 5.88 (AB, Jab = 9.5 Hz, 2H), 5.05-5.11 (m, 1H), 2.81-3.05 (m, 3H), 2.61-2.68 ( m, 4H, CH 2 CH 2 COOH), 2.14-2.17 (m, 1H). C13 NMR (300 MHz; CDCl3d6): 173; 171.1; 170.9; 169.1;

167; 134.9; 131.1; 124.1; 63.1; 49.3; 28.3; 31; 20.7.

Infrared (KBr pellet): υ O-H = 3210 cm -1; υ Aromatic C-H = 3084 cm -1; υ alkyl CH (symmetrical and asymmetric) = 2990 and 2851 cm "1; υ C = O imide (symmetrical and asymmetrical) = 1782 cm" 1 and 1711 cm "1; υ C = O imide ester = 1740; υ C = O acid = 1704; υ C = aromatic C = 1560 cm -1 and 1454 cm -1; CNC 138 0 cm -1.

d) Preparation of the prednisolone prodrug: 8S, 9S, 10R, 13S, 14S, 17R) -7,8,9,11,12,13,14,15,16,17-decay-11,17- dihydroxy-17- (2- (1- (hydroxymethylsuccinyl) -2,6-dioxopiperidin-3-yl) isoindoline-1,3-dione acetyl-10,13-

dimethyl-6H-cyclopenta [a] phenanthrene-3 (10H) -one (15)

10

-N OH

0 0 -O ^ 0 +

0 o

(14)

15

323 mg (0.83 mmol) of thalidomide derivative (3- (1,3-dihydro-1,3-dioxo-2H-isoindolyl-2-yl) -2,6-dioxo-piperidin-1-yl-metrl 1 ester) (-14) - have been dissolved, in -5 ml. —DichloromeLano. anhydrous, in .urr. 25 mL flask connected to a

20

25

30

tube with drying agent, forming a solution that was

kept at 0 ° C. 172 mg (0.83 ..... mmol) of

dicyclohexylcarbodiimide (DCC) and 11 mg (0.83 mmol) of dimethylaminopyridine (DMAP) were added to said solution. The mixture was kept under stirring at 0 ° C for 15-20 minutes. After this time, 3 to 5 drops of triethylamine and 300 mg (0.83 mmol) of prednisolone (4) were added to the solution. The mixture was kept under stirring for 6 hours and followed by thin layer chromatography until the termination of the reaction, where the mobile phase was a mixture of ethyl acetate and hexane in a ratio of 8: 2 and the stationary phase was composed of silica.

The reaction mixture was filtered and the filtrate was diluted with about 50 mL of dichloromethane. In a separatory funnel, the organic phase was washed 3 times with about mL of 0.02M bicarbonate solution. After this step, anhydrous sodium sulfate was added to the organic phase. The product was filtered and evaporated under reduced pressure and purified by column chromatography affording 250 mg of a white solid (prednisolone prodrug (15)) whose molecular formula is C39H42N2O2; MW = 730.76; MP: 205-210 ° C. The reaction yield was 41%.

1 H NMR (300MHz; CDCl 3 d): δ 7.89-7.87 (m, 2H); 7.75-7.78 (m; 2H); 7.32 (d; 1H;); 6.14 (d; 1H); 5.91 (s; 1H); 5.88 (s; 2H); 5.2 (s; 2H); 5.05 (m; 1H); 3.3 (s; 1H); 3.05-2.81 (m, 2HJ; 2.63-2.69 (m, 4H); 2.56 (d; 2H); 2.13-2.18 (2H; m); 1 1.8-2.1 (m; 7H); 1.43-1.49 (m; 4H); 1.36 (s; 3H); 0.9 (s; 3H) ppm.

- ■ -NMR-C13 - (300MHz; CDCl 3 d): δ 211.1; -171; 17.1.2;

170..8; ..... 169; . .167. 1, L5X._U__134. 9; 131.1; 128; 123.7;

120.1; 89.8; 69.2; 65.3; 63.1; 59.8; 50; 49.4; 45 ..4; 3 9. 7; 37; 33.3; 32.1; 31.1; 30.2; 29.7; 28.2; 22; 20.8; 19.8; 17.1 ppm.

Infrared (KBr pellet): υ O-H = 3210 cm -1; Aromatic C-H = 3084 cm -1; υ C-H alkyl (symmetrical and asymmetrical) = 2990 and 2851 cm -1; υ C = The imide (symmetrical and asymmetrical) = 1782 cm -1 and 1711 cm -1; υ C = The imide ester = 1740; C = Ocetone = 1707 cm-1; C = Aromatic C = 1560 cm-1 and 1454 cm-1; C-N-C 1380 cm-1.

2) Methodologies used for pharmacological assays and Results obtained.

a) Acetic acid induced abdominal writhing test The peripheral analgesic activity of the compounds was evaluated in the 0.6% acetic acid induced abdominal writhing model. In this assay, the animals are pretreated with the compounds of examples 1, 2, 3 or 4 orally at a concentration of 100 μπιοΐ / kg (0.1 mL / 20g) or with the standard drug dipyrone (vo; 100 μπιοΐ / kg (0.1 mL / 20g) In the negative control group, 5% arabic gum suspension is administered orally. After 1 hour of substance administration, an acetic acid solution 0 is injected intraperitoneally, 6% (0.1 ml / 10g mouse) and the number of writhes is recorded for 30 minutes.The decrease in the number of writhes is related to the peripheral analgesic activity of the compounds, and is expressed by the formula:

% analgesic activity = (n-n ') / 100n (1)

---------- n = _ average — of_number ___ of_containings __..... control group

(gum arabic. 5%); .........____ __

n '= mean number of test group writhing (compound)

Substance preparation

All substances were prepared in 5% gum arabic suspension at a dose of 100 μπιοΙ / kg. The 5% arabic gum showed no analgesic activity and had the same number of saline writhes (results not shown).

Statistical analysis

Significance levels between the experimental and control groups were made using the Tukey test. Values were considered significant when * P <0.05. Results were expressed as mean ± standard error of the mean. Results obtained

Table 1 indicates the number of abdominal writhing of acetic acid-induced mice and the percentage of protection by prednisolone and its derivatives (examples 1-4) at a concentration of 100 μπιοΐ / kg orally (n = 6, mean + standard deviation ).

Table 1

Treatment Number of writhing (average)% protection 1 \ control 48.5 + 2.3 - prednisolone 34 + 1.4 29, 9 Example 1 (prednisolone 5) 26.2 + 1.3 * 46 Example 2 (prednisolone 8) 31.3 + 2.2 * 35.5 Example 3 (prednisolone 11) 31.8 + 0.7 * 34.5 Example 4 (prednisolone 15) 30.5 + 2.7 * 37.1

Δ, Tab.el.a_l .. shows., That 'prednisolone'. at a dose of 100

10_ μΓηο ^ ί / ΐ ^^ presented a 29.9% inhibition percentage of 0.6% acetic acid-induced abdominal writhing. Examples 1-4 showed a variable protection percentage between 34.5-46%.

These data allow us to infer that the phthalimid subunit derived from thalidomide present in Examples 1-3 contributed to the investigated analgesic activity. In Example 4, thalidomide was derivatized to obtain a reciprocal prodrug joined by the presence of a spacer. In the latter case, the protection percentage was 37.1%, increasing the analgesic activity of prednisolone by 7% in this model.

b) Carrageenan-induced rat paw edema assay Injection of carrageenan into the Wistar rat paw causes edema and local inflammation which may be evidenced by increased rat paw thickness. Pretreatment with compounds having anti-inflammatory activity decreases edema and inflammation caused by carrageenan administration.

The compounds of the present invention were administered 1 hour prior to inoculation of the irritant (carrageenan) into the paws of animals. The follow-up of inflammation and anti-

The inflammatory reaction of the compounds of the invention was performed by measuring the rat paw thickness in millimeters.

The control group received subplantar, in the hind legs, the irritating chemical agent (carrageenan) and,

- —orally ", - saline solution. · - ----------- -

- ____________ The __, paws__ρ as.t.ex ior_es ._____ were measured_before

treatments and hourly,. for 6 hours after carrageenan administration by thickness gauge to determine carrageenan volumes (in mm). Results were expressed as the difference between paw readings before and after treatments.

Statistical analysis

Significance levels between the experimental and control groups were made using the Tukey test. * Values were considered significant when * P <0.05. Results were expressed as mean ± standard error of the mean.

Results Obtained Only the most active prednisolone-derived prodrug was evaluated for analgesic activity evaluation (Example 1) at a concentration of 100 μιηοΐ / kg.

Figure 2 shows the anti-inflammatory activity of prednisolone in the model used. Intraplantar animals were administered 0.1 mL carrageenan per paw at a concentration of 2 mg / mL. In the right paw only carrageenan was administered and in the left paw 0.1 ml of a 0.9% saline solution was also administered intraplantar. As a test, prednisolone 100 ymol / kg orally was administered 1 hour before carrageenan administration.

Figure 3 shows the anti-inflammatory activity of

____Example 1 on model used_ compared to carrageenan

(control inflammation) and prednisolone used as a control drug at a dose of 100 μπιοΐ / kg. As you can see, it is presented as anti-activity.

______ inXl_amatÁria_s ^ _gnifÁca.tiva_ when. „compared_a.„ carrageenan and

prednisolone used as a control drug. The antiinflammatory activity of example 1 is observed from the second hour onwards and shows a significant difference from prednisolone until the sixth hour. Figure 3 shows that the introduction of thalidomide-derived phthalimidic subunit increased the anti-inflammatory activity of the compound in the inflammation model used, this means that the greatest anti-inflammatory effect is possibly due to a synergism of action between the two subunits present in the compound. example 1 planned as a prodrug.

c) Gastric injury test

In order to characterize the gastric lesion profile caused by the new anti-inflammatory derivatives, the extent of the lesions caused by the administration of anti-inflammatory drugs was measured.

Gastric ulcerogenesis was verified in the same animals of the groups used for the paw edema model.

After 6 hours of paw readings, the animals were euthanized in CO2, and had their stomachs removed, opened in the direction of greatest curvature and washed with saline.

Through exposure of the mucosa, its color and integrity were observed. In case of lesions, they were counted and measured according to the gastric ulcerogenesis index (I.U.G.), which meets the criteria

for the classification of gastric mucosal lesions. Lesions smaller than 1 mm receive the number 1; at

------ lesions between 1.5 and 2.5 mm receive the number — 2;

____________ between .... 2, 5 and 3.5 mm receive the number_3; the lesions between 3, 5_ _

and 4,5 receive the number 4 and the lesions greater than 4,5 mm

receive the number 5. The results obtained were reported 20 as means ± Ε. Ρ M.

Statistical analysis

Significance levels between the experimental and control groups were made using the Tukey test. Values were considered significant when * P <0.05. Results were expressed as mean ± standard error of the mean.

Results obtained

In the gastric ulcerogenesis assay performed with the compound of example 1 no gastroulceration was observed. Common adverse effect on many anti-inflammatory drugs, especially those cyclooxygenase inhibitors.

The compounds of the invention may be administered in a variety of dosage forms, for example orally in the form of tablets, capsules, sugar or film-coated tablets, liquid solutions or suspensions; rectal route in the form of suppositories; parenterally, ie intramuscularly, or by intravenous and / or intrathecal and / or intraspinal infusion or injection.

The present invention also includes pharmaceutical compositions comprising such steroidal anti-inflammatory compounds, or pharmaceutically acceptable salts thereof, in combination with a pharmaceutically acceptable excipient (which may be a carrier or diluent).

Pharmaceutical compositions containing the compounds of

invention are "normally prepared by following methods"

-------------- Conventional — and — are — admired — in — form — fa-maeêu-t-i-Ga

appropriate. For example, solid oral pharmaceutical forms may contain together with the active compound diluents, that is: lactose, dextrose, sucrose, cellulose, cornstarch or potato starch; lubricants, ie: silica, talc, stearic acid, magnesium or calcium stearate, and / or polyethylene glycols; binding agents, for example: starches, gum arabic, gelatin, methylcellulose, carboxymethylcellulose or polyvinyl pyrrolidone; disintegrating agents, such as starch, alginic acid, starch or sodium starch alginates or glycolate; effervescent mixtures; dyes; sugars; wetting agents such as lectin, polysorbates, lauryl sulfates; and, generally, pharmacologically inactive and non-toxic substances used in pharmaceutical formulations. The pharmaceutical preparations of the present invention may be manufactured by mixing, granulating, tableting, sugar coating, or film coating processes, all already widely known and used.

Liquid dispersions for oral administration may be, for example, syrups, emulsions and suspensions. Syrups may contain as a carrier, for example, sucrose or sucrose with glycerine and / or manita and / or sorbitol. Suspensions and emulsions may contain as a carrier, for example, a natural gum, agar, sodium alginate, pectin, methylcellulose, carboxymethylcellulose, or alcohol.

polyvinyl. ^ __________.

Intramuscular injection suspensions or solutions may contain, together with the active compound, a pharmaceutically acceptable carrier, that is, sterile water, olive oil. _ _The. 1 and to _- .. de_e-tila, —g li cóis —, —this — is — p-ropi 1-eno — g-üc o i,, - and .se _.de.s_e_j.ado , associated with. an appropriate amount of lidocaine hydrochloride. Solutions for intravenous injections or infusions may contain as a carrier, for example sterile water or preferably they may be in the form of sterile, aqueous, isotonic saline or they may contain as propylene glycol carrier.

Suppositories may contain together with the active compound a pharmaceutically acceptable carrier, for example cocoa butter, polyethylene glycol, sorbitan polyoxyethylene, fatty acid ester surfactant or lecithin.

Claims (29)

<formula> formula see original document page 38 </formula> 1. Anti-inflammatory and immunosuppressive compounds, characterized in that they have the general formula: 0 Where: R = C6H4 (phenyl), (CH2) n where η = 1, 2, 3, 4, 5, 6, 7, 8 or 9, C5H3N C 4 H 3 N 2, C 3 H 2 N 3, NH, 2-hydroxyphenyl, 3-hydroxyphenyl, 4-hydroxyphenyl, 2-hydroxybenzyl, .3-hydroxybenzyl. . 4-hydroxybenzyl 1a, 4. 2-hydroxy-ethyl benzyl, 3-hydroxy-ethylbenzyl, 4-hydroxy-ethylbenzyl, benzyl, thiophene, furan, pyrrole, 2-hydroxy-pyridine, 3-hydroxy-pyridine, "" "4-hydroxy-pyridine , ..... "pyrazine, ..... -pi-r-im-i-di -, -— be-nzo-t-io-f-e - * - be-nzofur ano ^ - —I-ndo-1RT-quinoline, i quino 1 iia, - - therein allene ·, - - Ctt2 -? .- hydroxy-thiophene, ··· -QH2-3-hydroxy-Lyophene, C 1-2-hydroxy-furan, CH2-3-hydroxy-furan, CH2CH2-2-hydroxy-thiophene, CH2CH2-3-hydroxy-thiophene, CH2CH2-2-hydroxy-furan, CH2 CH2-3-hydroxy- furan, glutarimide, hydroxymethyl glutarimide, succinylhydroxymethylglutarimide, phthalylhydroxymethylphthalimide, 2-amino-phenyl, 3-amino-phenyl, 4-amino-phenyl, 2-amino-benzyl, 4-amino-benzyl, 2- amino-ethylbenzyl, 3-amino-ethylbenzyl, -4-amino-ethylbenzyl, · 2-amino-pyridine, 3-amino-pyridine, -4-amino-pyridine, CH2-2-amino-thiophene, CH2-3-amino -thiophene, CH2-2-amino-furan, CH2-3-amino-furan, CH2CH2-2-amino-thiophene, CH2CH2-3 -amino-thiophene, CH 2 CH 2 -2-amino-furan, CH 2 CH 2 -3-amino-furan or 3-methyl-2,6-dioxo-piperidin-1-yl-methyl-propyl ester; and W = H, (4 and / or 5 and / or 6 and / or 7) -F; (4 and / or 5 -and / or 6 and / or 7) -Cl; (4 and / or 5 and / or 6 and / or 7) -Br; (4 and / or 5 and / or 6 and / or 7) -NO 2; (4 and / or 5 and / or 6 and / or 7) -NH 2; (4 and / or 5 and / or 6 and / or 7) -OH; (4 and / or 5 and / or 6 and / or 7) -OCH 3; (4 and / or 5 and / or 6 and / or 7) -OCF3; (4 and / or 5 and / or 6 and / or 7) -CF 3. Anti-inflammatory compound according to claim 1, characterized in that W is hydrogen and R is CH2.1. Anti-inflammatory compound according to claims 1 and 2, characterized in that the compound is (8S, 9S, 10R, 13S, 14S, 17RJ.-7, 8, 9, .11, .12,13.14,15,15,16,17-decahydro-11,17-dihydroxy-17- (2- (1,3-dioxoisoindolin-2-yl) acetylacetate-10,13-dimethyl-6H -cyclopenta [a] --f-enanthrene-3 (1 OH) -one, this is prcdnisolor prodrug.a '_______ (5.L ._____________-_____: _: _______________ 4. An anti-inflammatory compound according to claim 1, wherein W is hydrogen and R is CeH4 in the para position. Anti-inflammatory compound according to claims 1 and 4, characterized in that the compound is (8S, 9S, 10R, 13S, 14S, 17R) -7,8,9,11,12,13 , 14,15,16,17-decahydro-11,17-dihydroxy-17- (4- (1,3-dioxoiso indolin-2-yl) benzoateacetyl-10,13-dimethyl-6H-cyclopenta [a] phenanthrene 3 (IOH) -one, that is prednisolone prodrug (8). Anti-inflammatory compound according to claim 1, characterized in that W is hydrogen and R is C6H4 in the meta position. Anti-inflammatory compound according to claims 1 and 6, characterized in that the compound is (8S, 9S, 10R, 13S, 14S, 17R) -7,8,9,11,12,13 , 14,15,16,17-decahydro-11,17-dihydroxy-17- (3- (1,3-dioxoiso indolin-2-yl) benzoateacetyl-10,13-dimethyl-6H-cyclopenta [a] phenanthrene 3 (IOH) -one ie prednisolone prodrug (H) · Anti-inflammatory compound according to claim 1, characterized in that W is hydrogen and R is 3-methyl-2,6-dioxo-piperidin-1-yl-methyl-propyl ester. Anti-inflammatory compound according to claim 1 and 8. characterized by the fact that the compound is 8S, 9S, 10R, 13S, 14S, 17R) -7,8,8,11,12,13,14,15,16,17-decahydro-11,17-dihydroxy- 17- (2- (1- (hydroxymethyl succinyl) -2,6-dioxopiperiperin-3-yl) isoindorin-1,3-dione acetyl-10,13-dimethyl-1,6H-cyclopen t a- [a -] - f-na-ntren © - ^ - (- I-OH-) -one, ie prednisolone prodrug (15). ■ - ■■ - - ---- --- r: ------- A process for preparing the compound as defined in claim 1 comprising the steps of: a) dissolving in a 25 mL flask the thalidomide derivative (A) in 5 mL of anhydrous dichloromethane; b) keeping the solution formed at 0 ° C; c) adding 172 mg (0.83 mmol) dicyclohexylcarbodiimide (DCC) and 11 mg (0.83 mmol) dimethylaminopyridine (DMAP) to said solution; d) stirring at 0 ° C the mixture obtained for 15-20 minutes; e) adding from 3 to 5 drops of triethylamine and 300 mg (0.83 mmol) of prednisolone to the solution; f) stirring the mixture obtained in step e) for a period of 3 to 6 hours and monitoring the reaction termination by thin layer chromatography, where the mobile phase is a mixture of ethyl acetate and hexane in a proportion ranging from 6 to 8: 2-4 and the stationary phase is silica; g) filtering the reaction mixture and diluting the filtrate with 50 mL of dichloromethane; h) Washing the organic phase 3 times with a separatory funnel with 15 ml of 0.02 M bicarbonate solution; i) adding anhydrous sodium sulfate to the organic phase; j) filtering, evaporating and purifying the product obtained in step i) to obtain a white solid which is prednisolone prodrug. A process according to claim 10, characterized in that when the derivative of --- fea-li-dominate ---- (- A -) - is —Θ — áe-gone-2 --- (- 1 -, - 3- ^ 1οχθί-3θ-ί-ηάο1-ί-η - 2 --- ί - 1-) t acetic (compound 3) The prednisolone prodrug — obtained will be the compound of claim 3. The process of claim 10, wherein when the thalidomide derivative (A) is 4- (1,3-dioxoisoindolin-2-yl) benzoic acid (compound 7) The prednisolone prodrug obtained will be the compound of claim 5. Process according to Claim 10, characterized in that when the thalidomide derivative (A) is 4- (1,3-dioxoisoindolin-2-yl) benzoic acid (compound 10) prednisolone drug obtained will be the compound of claim 7. Process according to Claim 10, characterized in that when the thalidomide derivative (A) is (3- (1,3-dihydro-1,3-dioxo-2H-isoindolyl-2-yl) -2,6-dioxopiperidin-1-ylmethyl] ester) (compound 14), the prednisolone prodrug obtained will be the compound of claim 9. Pharmaceutical composition comprising any of the compounds of claims 3, 5, 7 or 9 or pharmaceutically acceptable salts thereof in association with a pharmaceutically acceptable excipient. Pharmaceutical composition according to Claim 15, characterized in that it is in solid, liquid or suppository presentation forms. Pharmaceutical composition according to claim 15 or 16, characterized in that, when in solid presentation form, the "pharmaceutically acceptable excipient is a diluent", a lubricant, a dehydrating agent. bonding, -a-agent-des-ag-reg-ante-, -a-effervescent mixture, a dye, a sugar, or a wetting agent. Pharmaceutical composition according to claim 17, characterized in that the diluent is selected from the group consisting of lactose, dextrose, sucrose, cellulose, corn starch or potato starch. 19. Pharmaceutical composition of. Claim 17, characterized in that the lubricant is selected from the group consisting of silica, talc, stearic acid, magnesium or calcium stearate, and / or polyethylene glycols. Pharmaceutical composition according to Claim 17, characterized in that the binding agent is selected from the group consisting of starches, gum arabic, gelatin, methylcellulose, carboxymethylcellulose or polyvinyl pyrrolidone. Pharmaceutical composition according to claim 17, characterized in that the disintegrating agent is selected from the group consisting of starch, alginic acid or sodium or starch glycolate. Pharmaceutical composition according to claim 17, characterized in that the wetting agent is selected from the group consisting of lectin, polysorbates, lauryl sulfates. Pharmaceutical composition according to claim 15 or 16, characterized in that, when in liquid presentation form, the composition is a syrup, an emulsion or a suspension. ........... " 24. The composition of the invention, as claimed in claim 23, characterized in that: because the syrup contains as a carrier a sucrose or a sucrose with glycerin and / or manita and / or sorbitol. Pharmaceutical composition according to Claim 23, characterized in that the suspension and the emulsion contain as carrier a natural gum, agar, sodium alginate, pectin, methylcellulose, carboxymethylcellulose, polyvinyl alcohol or sterile aqueous saline. isotonic. Pharmaceutical composition according to Claim 25, characterized in that the carrier is still sterile water, olive oil, ethyl oleate, propylene glycol, optionally associated with lidocaine hydrochloride. Pharmaceutical composition according to Claim 15 or 16, characterized in that, when in the form of a suppository, the carrier is selected from the group consisting of cocoa butter, polyethylene glycol, sorbitan polyoxyethylene, surfactant. fatty acid or lecithin ester. Use of any of the compounds as defined in claims 3, 5, 7 or 9 for the preparation of a medicament for the treatment of chronic inflammatory processes. Use according to claim 28, characterized in that. whereas chronic inflammatory processes originate from a variety of diseases such as rheumatoid arthritis, lupus, psoriasis, Inflammatory Diseases, Linear "," "Inflammatory Eye Diseases", among others .—; —------- -------—