RU2772214C2 - Functionalized saccharides as anti-inflammatory agents - Google Patents

Abstract

FIELD: pharmacology; medicine.

SUBSTANCE: group of inventions relates to the field of pharmacology and medicine; it is intended for the effect on inflammation mechanisms in mammals. In particular, the use of a compound of the formula (Ib) is described, as well as a pharmaceutical composition based on this compound as an anti-inflammatory agent that affects inflammation mechanisms in animals. In addition, the group of inventions relates to a method (in vivo) for the suppression or inhibition of response inflammatory reactions by using the compound (Ib) and to a method for the suppression or inhibition of expression of at least one cytokine by injecting the compound (Ib). The present invention relates to compounds of functionalized saccharides used as anti-inflammatory agents that affect inflammation mechanisms in mammals.

EFFECT: providing an effective impact on inflammation mechanisms in mammals.

Formula (Ib)

11 cl, 8 tbl, 6 ex

Description

FIELD OF TECHNOLOGY TO WHICH THE INVENTION RELATES

The present invention relates to compounds of functionalized saccharides used as anti-inflammatory agents that affect the mechanisms of inflammation in mammals.

BACKGROUND OF THE INVENTION

Inflammation is a complex biological response of the immune system, aimed at protecting the body from substances harmful to health, from tissue damage and destruction. The inflammatory response can be initiated by physical, chemical, and infectious agents such as bacteria, viruses, and other pathogens. The purpose of inflammation is to eliminate the root cause of damage, to clean up damaged cells and tissues, and to initiate tissue repair.

The inflammation can be acute or chronic. Acute inflammation refers to the initial response of the body's immune system to harmful irritants. Chronic inflammation is a form of long-term inflammation that leads to the recruitment of macrophages and T-lymphocytes. Macrophages are large phagocytic leukocytes, and together with T-lymphocytes, which are known to produce cytokines and enzymes, they cause longer-term cell damage. Although it is essential for the body to have a properly functioning immune system, it is nevertheless important to control these inflammatory mechanisms, as chronic inflammation can lead to various pathologies and even cancer.

However, the term "inflammation" is not synonymous with the term "infection". Infection refers merely to the interaction of a microorganism invasion with the host's inflammatory response to that invasion. On the other hand, inflammation describes an inflammatory mechanism that occurs independently of the pathogen. Despite this, in medicine, inflammation or organ-specific inflammation is often referred to as infection because microbial invasion is commonly seen in association with inflammation. However, it is important to distinguish between the two because many pathological conditions involve inflammatory processes that are not due to microbial or viral invasion, and conversely, not all microbial or viral infections result in inflammation.

The confusion between the concepts of infection and inflammation is especially characteristic in cases of gastritis or the presence of inflammation in the stomach. Many doctors consider chronic gastritis to be the equivalent of a Helicobacter pylori (H. pylori) infection. However, it is known that there are many other causes of this disease. In addition, gastric pathology also persists even after successful eradication of H. Pylori. In a publication by Genta et al., American Journal of Gastroenterology, it was most recently reported that approximately 20% of patients from whom a gastrobiopsy was taken were diagnosed with gastritis not associated with Helicobacter pylori infection. Given its prevalence, in the publication of Borkman et al., New England Journal Of Medicine, it was proposed to consider gastritis not associated with Helicobacter pylori infection as a separate nosological form. Known causes of stomach inflammation in the absence of a bacterial infection, particularly H. pylori infection, include:

(i) viral infection (cytomegalovirus, herpes simplex);

(ii) chemical or reactive inflammation caused by reflux of bile and pancreatic juice into the stomach or by exogenous substances such as NSAIDs (non-steroidal anti-inflammatory drugs), ASAs (acetylsalicylic acid), alcohol or chemotherapy drugs;

(iii) autoimmune inflammation characterized by the presence of antibodies against parietal cells;

(iv) radiation and eosinophilic inflammation;

(v) inflammation during collagenosis;

(vi) inflammation in Crohn's disease;

(vii) inflammation in sarcoidosis;

(viii) stress-induced inflammation;

(ix) human gastric bacterial species other than H. pylori Helicobacter (NHPH), of which H. suis is the most common. They are associated with chronic gastritis, gastric ulcer and other pathological changes in the stomach, both in animals and humans (G. Zhang et al., Vet Res (2016) 47: 101, p. 1 1. 1-3);

(x) idiopathic: all cases of gastritis in which no cause can be identified.

In addition to the problem of non-Helicobacter gastritis, gastric ulcer recurrence remains a problem in the treatment of humans and farm animals, even after successful eradication of Helicobacter bacteria. For example, in the case of pigs, although ulcer scarring may occur in treated animals, ulcer recurrence is still a common problem. A recurrence of peptic ulcer causes discomfort and loss of appetite in the animal, leading to weight loss and negative economic consequences. But even in the treatment of people, the recurrence of stomach ulcers is a serious threat to health. Currently, patients suffering from chronic gastritis and/or gastric ulcers are forced to use long-term proton pump inhibitors (PPI) to block the formation of acid in the stomach. It is known that this long-term use of proton pump inhibitors is accompanied by many side effects and increases the risk of developing chronic kidney disease, renal failure and an increased risk of cardiovascular complications.

Alternatively, instead of blocking stomach acid production, gastric ulcers and gastritis are treated by targeting Helicobacter pylori bacteria.

In addition, the use of non-steroidal anti-inflammatory drugs (NSAIDs) is contraindicated to alleviate the state of inflammation of the stomach, since these drugs are known to be a risk factor for the occurrence of gastric pathology.

Based on the foregoing, it can be concluded that there is a need for anti-inflammatory drugs, as a result of which the disadvantages of modern methods of anti-inflammatory treatment can be overcome, such as a large number of side effects, namely an increased risk of chronic kidney disease, renal failure, increased the risk of cardiovascular complications and other similar negative side effects, limited scope, non-compliance by the patient with instructions for taking the drug, and as a result of the use of which problems of ulcer recurrence, in particular, recurrence of gastric ulcer, can be overcome.

SUMMARY OF THE INVENTION

To the surprise of the applicant of the present invention, it turned out that it is possible to create anti-inflammatory drugs that would satisfy the requirements mentioned above.

The aim of the present invention is to provide an alternative anti-inflammatory agent instead of non-steroidal anti-inflammatory drugs (NSAIDs) to eliminate inflammation in the stomach of mammals suffering from gastritis, gastric ulcer, or mammals at risk of developing these diseases.

The aim of the present invention is to provide an alternative to the long-term use of proton pump inhibitors (PPIs) in subjects suffering from gastric ulcers and chronic gastritis.

The aim of the present invention is to provide an anti-inflammatory agent that can act on the inflammatory mechanisms underlying gastric ulcer and gastritis in mammals.

The present invention relates to compounds useful as an anti-inflammatory agent that act on the mechanisms of inflammation in mammals [hereinafter referred to as anti-inflammatory compound (C)], wherein said anti-inflammatory compound (C) is at least one of the compounds represented by the general formulas (I), (II), (III) or (IV), or a pharmaceutically acceptable salt, pharmaceutically acceptable solvate, isomer or mixture thereof:

formula (I)

formula (II)

where

- each of R ¹ is independently selected from -OH or -N(R ^1a )(R ^1b ), where each of R ^1a and R ^1b , independently of each other in each case of their presence, is selected from the group consisting of H, protective groups for amino group, aryl, heteroaryl, aliphatic and heteroaliphatic group,

- each of R ² , R ⁴ , R ⁵ , R ¹¹ and R ¹² , independently of each other in each case of their presence, is selected from the group consisting of H, -OR ^z , -N(R ^z ) ₂ , aryl, a heteroaryl, aliphatic, and heteroaliphatic group, wherein R ^z is independently selected from the group consisting of H, a hydroxyl protecting group, an amino protecting group, an aryl, heteroaryl, aliphatic, heteroaliphatic, and carbohydrate moiety,

- each R ³ is independently selected from the group consisting of H, -OH, -NH ₂ , -SH, OR ^w , -NH(R ^w ), -N(R ^w ) ₂ , -SR ^w , -O(C =O)R ^w , -NH(C=O)R ^w , -O(C=NH)R ^w , -NH(C=NH)R ^w , -S(C=NH))R ^w , -NH( C=S)R ^w , -S(C=O)R ^w , -O(C=S)R ^w , -S(=S)R ^w , aryl, heteroaryl, aliphatic and heteroaliphatic group, where R ^w is selected from a group consisting of a carbohydrate fragment, an aryl, heteroaryl, aliphatic and heteroaliphatic group,

- each of R ⁶ , R ¹⁴ and R ¹⁵ , independently of each other and in each case of their presence, is selected from the group consisting of H, a protective group for a hydroxyl group, an aryl, heteroaryl, aliphatic, heteroaliphatic group and a carbohydrate moiety,

- each R ⁷ is independently selected from the group consisting of H, C(=O)N(R ^z' ) ₂ , -C(=O)OR ^z' , hydroxyl protecting group, aryl, heteroaryl, aliphatic, heteroaliphatic a group and a carbohydrate moiety, wherein R ^z' is independently selected from the group consisting of H, a hydroxyl protecting group, an amino protecting group, an aryl, a heteroaryl, an aliphatic, and a heteroaliphatic group,

- each R ¹⁰ is independently selected from the group consisting of -C(=O)N(R ^l )(R ^p ), -C(=O)OR ^k and -CH ₂ OR ^k where R ^l and R ^p are independently from each other and in each case of their presence, is selected from the group consisting of H, a protective group for an amino group, an aryl, heteroaryl, aliphatic, heteroaliphatic group and a carbohydrate fragment, where R ^k is independently selected from the group consisting of H, a protective group for hydroxyl group, aryl, heteroaryl, aliphatic, heteroaliphatic group and carbohydrate moiety,

- each of R ¹³ is independently selected from the group consisting of -OH and -N(R ^o'' )(R ^m'' ), where R ^o'' and R ^m'' , independently of each other in each case of their presence , is selected from the group consisting of H, -C(=O)R ^h , a protective group for the amino group, aryl, heteroaryl, aliphatic and heteroaliphatic group, where R ^h is selected from the group consisting of carbohydrate moiety, aryl, heteroaryl, aliphatic and heteroaliphatic group,

- each of (R ^o ) and (R ^m ), independently of each other in each case of their presence, is selected from the group consisting of H, -C(=O)R ^w' , a protective group for an amino group, aryl, heteroaryl, aliphatic and heteroaliphatic groups, where R ^w' is selected from the group consisting of carbohydrate fragment, aryl, heteroaryl, aliphatic and heteroaliphatic group;

- each of (R ^s ) and (R ^t ), independently of each other and in each case of their presence, is selected from the group consisting of H, -C(=O)R ^w'' , a protective group for an amino group, aryl, heteroaryl, aliphatic and heteroaliphatic groups, where R ^w'' is selected from the group consisting of carbohydrate fragment, aryl, heteroaryl, aliphatic and heteroaliphatic group;

- each of R ^6a and R ^6a' , independently of each other and in each case of their presence, is selected from H or -OH, preferably R ^6a is H and R ^6a' is -OH;

- each R ^c is independently selected from the group consisting of H, halogen, heteroaryl, -OR ^q , -N(R ^q ) ₂ , -SR ^q , NO ₂ , -NC, -CN, -N ₃ , -N( R ^q )=NR ^q , -CHO, -C(=O)R ^q , -C(=S)R ^q , C(=NR ^q )R ^q , -C(=O)OR ^q , -C(= NR ^q )OR ^q , -C(=NR ^q )N(R ^q ) ₂ , -C(=O)N(R ^q ) ₂ , -C(=S)OR ^q , -C(=O)SR ^q , -C(=S)SR ^q , -P(=O)(OR ^q ) ₂ , -S(=O)(OR ^q ), -S(=O) ₂ (OR ^q ), -P(=O )N(R ^q ) ₂ , -P(=O) ₂ N(R ^q ) ₂ , -C(=O)NR'S(=O) ₂ R ^q , -S(=O)N(R ^q ) ₂ and -S(=O) ₂ N(R ^q ) ₂ , where each R ^q is independently selected from the group consisting of H, an aliphatic, heteroaliphatic group, an aryl, a heteroaryl group, and a protecting group for a hydroxyl group,

- each R ⁱ is independently selected from the group consisting of H, a protecting group for a hydroxyl group, an aliphatic, a heteroaliphatic group, an aryl and a heteroaryl,

- each Lipid is independently selected from an H or C _1-30 aliphatic moiety in which 0 to 10 methylene units are optionally replaced by -O-, -NR ^x -, -S-, -C(=O)-, -C( =NR ^x )-, -S(=O)-, -S(=O) ₂ -, -N=N-, -C=N-, -C(R ^y )=C(R ^y' )-, -N-O-, arylene, or heteroarylene moiety, wherein R ^x is each independently selected from the group consisting of H, an aliphatic, heteroaliphatic, aryl, heteroaryl, or amino protecting group, and wherein R ^y and R ^y' are each , independently of each other and in each case of their presence, is selected from the group consisting of H, aliphatic, heteroaliphatic, aryl and heteroaryl groups.

In addition, the present invention relates to a pharmaceutical composition for use as an anti-inflammatory agent that affects the mechanisms of inflammation in mammals, which comprises a carrier and, as an active ingredient, an effective amount of an anti-inflammatory compound (C) as defined in any one of the inventions. implementation options.

In addition, the present invention relates to a method for preparing said pharmaceutical composition.

In addition, the present invention relates to an anti-inflammatory compound (C) defined in any one of the embodiments presented in the invention for use as a drug.

In addition, the present invention relates to a pharmaceutical composition comprising a carrier and, as an active ingredient, an effective amount of an anti-inflammatory compound (C) defined in any one of the embodiments presented in the invention, for use as a medicine.

In addition, the present invention relates to an anti-inflammatory compound (C) as defined in any one of the embodiments of the invention for use in the treatment of inflammatory diseases or inflammation-related diseases.

In addition, the present invention relates to an anti-inflammatory compound (C) as defined in any one of the embodiments of the invention for use in the treatment of a disease mediated by at least one cytokine.

In addition, the present invention relates to an in vivo method for suppressing or inhibiting inflammatory responses by using an anti-inflammatory compound (C) defined in any one of the embodiments presented in the invention.

In addition, the present invention relates to a method for suppressing or inhibiting the expression of at least one cytokine in a mammal.

In addition, the present invention relates to a method of treating inflammation-related diseases associated with cytokine expression levels in mammals.

ANTI-INFLAMMATORY COMPOUND (C) AND ITS PHARMACEUTICALLY ACCEPTABLE SALT

Further, it should be understood that all definitions and advantages described above for the anti-inflammatory compound (C) apply equally to this embodiment and to all additional embodiments described below.

Used above and below in the invention, the definitions have the following meanings, unless otherwise indicated.

The term "aliphatic" includes both saturated and unsaturated, non-aromatic straight chain (i.e. straight), branched, acyclic and cyclic hydrocarbon chain having from 1 to 30 carbon atoms, which are optionally substituted with one or more groups including, but not limited to, alkyl, alkynyl, alkenyl, aryl, halide, nitro, amino, ether, ketone, aldehyde, hydroxy, carboxylic acid, or alkoxy. In particular embodiments, the C _F-G aliphatic group used in the invention defines a C _F-G aliphatic group having F to G carbon atoms, e.g., C _1-12 aliphatic group defines an aliphatic group having 1 to 12 carbon atoms, C _1-10 aliphatic group defines an aliphatic group containing 1 to 12 carbon atoms.

It will be apparent to one of ordinary skill in the art that "aliphatic" as used herein includes, but is not limited to, alkyl, alkenyl, and alkynyl moieties. As used herein, the terms "alkyl", "alkenyl", and "alkynyl" have a wide range of conventional meanings and may include a moiety that is linear, branched, cyclic (cyclooalkyl, cycloalkenyl, cycloalkynyl) moieties, or a combination thereof.

The term "alkyl", used alone or in combination, denotes an alkane-derived radical which may be linear alkyl, branched alkyl or cyclic alkyl containing from 1 to 30 carbon atoms, unless otherwise indicated. A straight or branched alkyl group is attached at any available point to form a stable compound. Alkyl also includes a linear or branched alkyl group that contains a cycloalkyl moiety or is interrupted by a cycloalkyl moiety. In specific embodiments, C _A-B alkyl denotes a linear or branched alkyl radical having from A to B carbon atoms, for example C _1-15 alkyl denotes a linear or branched alkyl radical having from 1 to 15 carbon atoms, C _1-12 alkyl denotes a linear or branched alkyl radical having 1 to 12 carbon atoms, C _1-6 alkyl denotes a linear or branched alkyl radical having 1 to 6 carbon atoms, such as, for example, methyl, ethyl, 1-propyl, 2 -propyl, 1-butyl, 2-butyl, 2-methyl-1-propyl. In specific embodiments, cyclic C _C-D alkyl refers to a cyclic alkyl radical having C to D carbon atoms, such as C _3-6 cyclic alkyl.

The term "alkenyl", used alone or in combination, means a linear or branched hydrocarbon containing 1-30 carbon atoms, unless otherwise indicated, and at least one carbon-carbon double bond. Examples of alkenyl groups include ethenyl, propenyl, isopropenyl, butenyl, cyclohexenyl, cyclohexenylalkyl, and other similar alkenyl groups. Alkenyl also includes a linear or branched alkenyl group that contains a cycloalkyl moiety or is interrupted by a cycloalkyl moiety. The carbon-carbon double bonds may be present either in the cycloalkyl portion or in the straight chain or branched portion. In specific embodiments, C _H-I alkenyl is a linear or branched alkenyl radical having H to I carbon atoms, for example, C _1-6 alkenyl is a linear or branched alkenyl radical having 1 to 6 carbon atoms.

The term "alkynyl", used alone or in combination, means a linear, branched or cyclic hydrocarbon containing from 1 to 30 carbon atoms containing at least one carbon-carbon triple bond. Examples of alkynyl groups include ethynyl, propynyl, butynyl, and the like.

As used herein, the term "heteroaliphatic" refers to an aliphatic moiety as defined herein, which includes both saturated and unsaturated, non-aromatic, linear (i.e., unbranched), branched, acyclic, cyclic (i.e., heterocyclic), or polycyclic hydrocarbons having from 1 up to 30 carbon atoms, unless otherwise indicated, which are optionally substituted with one or more groups including, but not limited to, alkyl, alkynyl, alkenyl, aryl, halide, nitro, amino, ether, ketone, aldehyde, hydroxyl groups , a carboxylic acid group or an alkoxy group, and which contain one or more heteroatoms such as oxygen, sulfur, nitrogen, phosphorus or silicon atoms, for example, instead of carbon atoms. In specific embodiments, the heteroaliphatic moieties are substituted by independently replacing one or more of their hydrogen atoms with one or more substituents. For a person of ordinary skill in this field, it is obvious that "heteroaliphatic", as intended in the invention, includes heteroalkyl, heterocycloalkyl, heterocycloalkenyl and heterocycloalkynyl fragments. Therefore, the term "heteroalkyl" as used herein includes linear, branched and cyclic alkyl groups which contain one or more heteroatoms such as oxygen, sulfur, nitrogen, phosphorus or silicon, for example, instead of carbon atoms. A similar definition applies to other nomenclature terms such as "heteroalkenyl", "heteroalkynyl" and other similar terms. In addition, as used herein, the terms "heteroalkyl", "heteroalkenyl", "heteroalkynyl", and the like, encompass both substituted and unsubstituted groups.

As used herein, the term "aryl", used alone or in combination, means any aromatic group based on carbon atoms, including, but not limited to, phenyl, tolyl, xylyl, cumenyl, naphthyl, anthracenyl, and the like, optionally fused with cycloalkyl or heterocyclyl containing preferably 5-7, more preferably 5-6 ring atoms and/or optionally substituted with 1 to 5 groups or substituents. Aryl may be optionally substituted with one or more groups including, but not limited to, alkyl, alkynyl, alkenyl, aryl, halide, nitro, amino, ether, ketone, aldehyde, hydroxyl, carboxylic acid, or alkoxy groups, provided that the substituent is attached at one point to the aryl, or provided that the substituent is attached at two points to the aryl, to form a bicyclic system, for example, benzodioxole, benzodioxane, benzimidazole.

The term "heteroaryl", used alone or in combination, means a monocyclic aromatic ring structure containing 5 or 6 ring atoms, or a bicyclic aromatic group having 8 to 10 atoms, containing one or more, preferably 1-4, more preferably 1-3 heteroatoms independently selected from the group O, S and N and optionally substituted with 1 to 5 groups or substituents including, but not limited to, alkyl, alkynyl, alkenyl, aryl, halide, nitro, amino , an ether, a ketone, an aldehyde, a hydroxyl group, a carboxylic acid group, or an alkoxy group. It is contemplated that heteroaryl also includes oxidized S or N forms such as sulfinyl, sulfonyl and N-oxide of the tertiary nitrogen atom in the ring. A carbon or nitrogen atom is attached to the heteroaryl ring structure in such a way that the stability of the aromatic ring is maintained. More specifically, the term "heteroaryl" includes, but is not limited to, pyridyl, furanyl, thiophenyl, thiazolyl, isothiazolyl, triazolyl, imidazolyl, isoxazolyl, pyrrolyl, pyrazolyl, pyrimidyl, benzofuranyl, isobenzofuranyl, benzothiazolyl, benzoisothiazolyl, benzotriazolyl, indolyl, isoindolyl, benzoxazolyl, quinolyl, isoquinolyl, benzimidazolyl, benzisoxazolyl, benzothiophenyl, dibenzofuran and benzodiazepin-2-on-5-yl; and other similar heteroaryl groups.

According to a specific embodiment of the present invention, Lipid is an unsaturated C _4-20 hydrocarbon chain in which 1 to 10 methylene units are replaced by -C(R ^y )=C(R ^y' )-, where each of R ^y and R ^y' , independently of each other in each case of their presence, is selected from the group consisting of H, aliphatic, heteroaliphatic, aryl or heteroaryl group; and wherein said hydrocarbon chain is optionally substituted with C _1-6 alkyl, C _3-6 cyclic alkyl, C _1-6 alkenyl or hydroxyl group, preferably Lipid is a C _4-20 hydrocarbon chain wherein 1 to 5 methylene units are replaced by -C(R ^y )=C(R ^y' )-, where each of R ^y and R ^y' , independently of each other in each case of their presence, is selected from the group consisting of H, C _{1– 6} alkyl and C _1-6 alkenyl, and wherein said hydrocarbon chain is optionally substituted with C _1-6 alkyl, C _3-6 cyclic alkyl, C _1-6 alkenyl, or a hydroxyl group, more preferably Lipid is selected from moieties having the following formulas:

Even more preferably, the Lipid is selected from moieties having the following formulas:

According to a specific embodiment of the present invention, Lipid is a saturated hydrocarbon chain having a common Z:

Formula Z

where

- each of R ^9' and R ^9'' , independently of each other and in each case of their presence, is selected from the group consisting of H, -OH and C _1-6 alkyl,

- each of R ^x' and R ^x'' , independently of each other and in each case of their presence, is selected from H, C _1-15 alkyl and aryl, where the specified aryl is optionally additionally substituted with C _1-15 alkyl, and where n=4-30, and m=0 or 1

Preferably, the saturated hydrocarbon chain is selected from moieties having the following formulas:

According to a specific embodiment of the present invention, R ¹ is independently selected from -OH or -N(R ^1a )(R ^1b ), where each of R ^1a and R ^1b , independently of each other in each case of their presence, is selected from the group consisting of H, C _1-20 aliphatic and C _1-20 heteroaliphatic, preferably R ¹ is OH, NH ₂ , or NH(R ^1b ), where R1b is a C _1-10 cycloalkyl group or C _1-10 cycloalkenyl which is optionally substituted with halogen, hydroxyl or alkoxy, more preferably R ¹ is -NH(R ^1b ), where R ^1b has the following formula:

According to a particular embodiment of the present invention, R ³ is independently selected from the group consisting of H, -OH, and -OR ^w , where R ^w is selected from the group consisting of a carbohydrate moiety, wherein said carbohydrate moiety is independently selected from the group consisting of D- and L-monosaccharides, such as, in particular, D-erythrose, L-erythrose, D-threose, L-threose, L-erythrulose, D-erythrulose, D-arabinose, L-arabinose, D-deoxyribose, L-deoxyribose , D-lyxose, L-lyxose, D-ribose, L-ribose, D-ribulose, L-ribulose, D-xylose, L-xylose, D-xylulose, L-xylulose, D-allose, L-allose, D -altrose, L-altrose, D-galactose, L-galactose, D-glucose, L-glucose, D-gulose, L-gulose, D-idose, L-idose, D-mannose, L-mannose, D-talose , L-talose, D-fructose, L-fructose, D-psicose, L-psicose, D-sorbose, L-sorbose, D-tagatose, L-tagatose, D-fucose, L-fucose, D-rhamnose and L -rhamnose, disaccharides such as in particular sucrose, lactose, trehalose and maltose, trisaccharides such as in particular a carbose, raffinose and melicitose, more preferably R ³ is -OR ^w , where R ^w is a D-monosaccharide, such as, in particular, D-erythrose, D-threose, D-erythrulose, D-arabinose, D -deoxyribose, D-lyxose, D-ribose, D-ribulose, D-xylose, D-xylulose, D-allose, D-altrose, D-galactose, D-glucose, D-gulose, D-idose, D-mannose , D-talose, D-fructose, D-psicose, D-sorbose, D-tagatose, D-fucose and D-rhamnose, even more preferably R ³ is -OR ^w , where R ^w is independently selected from the group consisting of from D-glucose, D-galactose, D-allose, D-altrose, D-mannose, D-idose, D-halose or D-talose, most preferably R ³ is -OR ^w where R ^w is D-glucose.

According to a specific embodiment of the present invention, each of R ² , R ⁴ , R ⁵ , R ¹¹ and R ¹² , independently of each other in each case of their presence, is selected from the group consisting of H, OR ^z , C _1-12 aliphatic , C _1-12 heteroaliphatic and carbohydrate fragment, where R ^z is selected from the group consisting of H, a protective group for the hydroxyl group, C _1-12 aliphatic group, C _1-12 heteroaliphatic group, aryl and heteroaryl, and where the specified carbohydrate fragment selected from the group consisting of D-monosaccharides, such as, in particular, D-erythrose, D-threose, D-erythrulose, D-arabinose, D-deoxyribose, D-lyxose, D-ribose, D-ribulose, D- xylose, D-xylulose, D-allose, D-altrose, D-galactose, D-glucose, D-gulose, D-idose, D-mannose, D-talose, D-fructose, D-psicose, D-sorbose D -tagatose, D-fucose and D-rhamnose, preferably each R ² , R ⁴ , R ⁵ , R ¹¹ and R ¹² , independently of each other in each case of their presence, is selected from the group consisting from H, OR ^z , a hydroxyl protecting group, C _1-6 alkyl, and a carbohydrate moiety, wherein R ^z is selected from the group consisting of H, a hydroxyl protecting group, and C _1-6 alkyl, and wherein said carbohydrate moiety is selected from the group consisting of D-glucose, D-galactose, D-allose, D-altrose, D-mannose, D-idose, D-halose and D-talose, more preferably R ² , R ⁴ , R ¹¹ and R ¹² , independently of one another and in each case present, is selected from H or OR ^z , where R ^z is selected from H or C _1-6 alkyl, more preferably R ⁵ is C _1-6 alkyl, even more preferably R ² is H, R ⁴ , R ¹¹ and R ¹² is -OH and R ⁵ is -CH ₃ .

According to a specific embodiment of the present invention, each of R ⁶ , R ¹⁴ and R ¹⁵ , independently of each other in each case of their presence, is selected from the group consisting of H, a protecting group for a hydroxyl group, C _1-12 aliphatic and C _{1 -12} heteroaliphatic group, preferably R ⁶ , R ¹⁴ and R ¹⁵ , independently of each other in each occurrence, are selected from the group consisting of H or a protecting group for a hydroxyl group, more preferably R ⁶ , R ¹⁴ and R ¹⁵ , independently of each other and in each case of their presence, were H.

According to a specific embodiment of the present invention, R ⁷ is a -C(=O)N(R ^z' ) ₂ group, where each R ^z' is independently selected from the group consisting of H, aryl, heteroaryl, and C _1-12 alkyl , preferably each R ^{z' is} independently selected from H or C _1-12 alkyl, more preferably each R ^{z' is} independently selected from the group consisting of H, methyl, ethyl and propyl, most preferably each R ^z' is independently H.

According to a particular embodiment of the present invention, each R ¹⁰ is independently selected from -C(=O)N(R ^l )(R ^p ) or -C(=O)OR ^k , where R ^l and R ^p are independently and in each case present, is selected from the group consisting of H, an amino protecting group, aryl, heteroaryl, C _1-12 aliphatic, and C _1-12 heteroaliphatic, and where R ^k is independently selected from the group consisting of H, a protecting group for hydroxyl, aryl, heteroaryl, C _1-30 aliphatic and C _1-30 heteroaliphatic, preferably each R ¹⁰ is independently selected from -C(=O)N(R ^l )(R ^p ) or - C(=O)OR ^k , where R ^l and R ^p , independently of each other and in each case of their presence, are selected from the group consisting of H, an amino protecting group, aryl, heteroaryl and C _1-6 alkyl, where each R ^k is independently selected from the group consisting of H, a protecting group for a hydroxyl group and C _1-6 alkyl, more preferably each R ¹⁰ is not independently selected from -C(=O)N(R ^l )(R ^p ) or -C(=O)OR ^k , where R ^l and R ^p , independently of each other in each case of their presence, are selected from the group consisting of from H, an amino protecting group, methyl, ethyl and propyl, wherein each R ^k is independently selected from the group consisting of H, a hydroxyl protecting group, methyl, ethyl and propyl, most preferably R ¹⁰ is -C (=O)NH ₂ or -C(=O)OH.

According to a specific embodiment of the present invention, each R ¹³ is independently selected from the group consisting of -OH and -N(R ^o'' )(R ^m'' ), where R ^o'' and R ^m'' , independently of each other each other in each occurrence is selected from the group consisting of H, -C(=O)R ^h , amino protecting group, C _1-12 aliphatic and C _1-12 heteroaliphatic, where R ^h is independently selected from C _{1 -12} aliphatic or C _1-12 heteroaliphatic group, preferably R ^o'' and R ^m' , independently of each other in each occurrence, are selected from the group consisting of H, -C(=O)R ^h and C _1-12 alkyl, where R ^h is C _1-12 alkyl, more preferably R ^o'' and R ^m'' , independently of each other in each occurrence, are selected from the group consisting of H, - C(=O)R ^h and C _1-6 alkyl, where R ^h is C _1-6 alkyl, most preferably R ^o'' is H and R ^m' is -C(=O)CH ₃ .

According to a specific embodiment of the present invention, each of (R ^o ) and (R ^m ), independently of each other in each case of their presence, is selected from the group consisting of H, -C(=O)R ^w' , a protective group for the amino group , C _1-12 aliphatic and C _1-12 heteroaliphatic, where R ^w' is independently selected from the group consisting of C _1-12 aliphatic and C _1-12 heteroaliphatic, preferably (R ^o ) and (R ^m ) , independently of each other in each occurrence, were selected from the group consisting of H, -C(=O)R ^w' , an amino protecting group, and C _1-12 alkyl, where R ^w' is C _1-12 alkyl, more preferably (R ^o ) and (R ^m ), independently of each other in each case of their presence, are selected from the group consisting of H, -C(=O)R ^w' and C _1-6 alkyl, where R ^w' is C _1-6 alkyl, most preferably (R ^o ) is H and (R ^m ) is -C(=O)CH ₃ .

According to a specific embodiment of the present invention, each of (R ^s ) and (R ^t ), independently of each other and in each case of their presence, is selected from the group consisting of H, -C(=O)R ^w'' , protective groups for amino, C _1-12 aliphatic and C _1-12 heteroaliphatic, where R ^w'' is independently selected from C _1-12 aliphatic or C _1-12 heteroaliphatic, preferably (R ^s ) and (R ^t ) , independently of each other in each occurrence, were selected from the group consisting of H, -C(=O)R ^w'' , an amino protecting group, and C _1-12 alkyl, where R ^w'' is C _{1 –12} alkyl, more preferably (R ^s ) and (R ^t ), independently of each other in each occurrence, are selected from the group consisting of H, -C(=O)R ^w'' and C _{1– 6} alkyl, where R ^w'' is C _1-6 alkyl, most preferably (R ^s ) is H and (R ^t ) is -C(=O)CH ₃ .

According to a particular embodiment of the present invention, each R ^c is independently selected from the group consisting of H, -C(=O)OR ^q , -C(=O)SR ^q , -C(=S)OR ^q , -C( =O)SR ^q , -C(=S)SR ^q , where each R ^q is independently selected from the group consisting of H, C _1-12 alkyl, aryl, heteroaryl and a hydroxyl protecting group, preferably each R ^c is independently selected from the group consisting of H, -C(=O)OR ^q , -C(=O)SR ^q and -C(=O)SR ^q , where each of R ^q is independently selected from the group consisting of H, C _1-6 alkyl and a protecting group for a hydroxyl group, more preferably each R ^c is independently selected from H or -C(=O)OR ^q where each R ^q is independently selected from the group consisting of H, methyl, ethyl and propyl, most preferably R ^c is -C(=O)OH.

According to a specific embodiment of the present invention, each R ⁱ is independently selected from the group consisting of H, a protecting group for a hydroxyl group, C _1-12 aliphatic, C _1-12 heteroaliphatic, aryl and heteroaryl, preferably R ⁱ is independently selected from the group consisting of H, a hydroxyl protecting group, C _1-12 alkyl, aryl and heteroaryl, more preferably R ⁱ is independently selected from the group consisting of H, a hydroxyl protecting group and C _1-6 alkyl, even more preferably R ⁱ is independently selected from the group consisting of H, a hydroxyl protecting group, methyl, ethyl and propyl, most preferably R ⁱ is H.

The anti-inflammatory compounds (C) of the present invention described in detail above have multiple centers of chirality and exist in stereochemically isomeric forms. As used herein, the term "stereochemically isomeric forms" refers to all possible compounds consisting of the same atoms, linked by the same sequence of chemical bonds, but having different spatial structures that are not equivalent, which the anti-inflammatory compounds specified in the invention may have. (C).

Unless otherwise mentioned or indicated, the chemical designation of the above detailed anti-inflammatory compounds (C) embraces the mixture of all possible stereochemically isomeric forms that said anti-inflammatory compounds (C) may have. Said mixture may contain all diastereomers and/or enantiomers of the basic molecular structure of said anti-inflammatory compounds (C). It is intended that all stereochemically isomeric forms of the compounds of the present invention, either in pure form or in admixture with each other, are within the scope of the present invention.

Preferred anti-inflammatory compounds (C) are compounds of formula (Ia) [hereinafter referred to in the invention as class I anti-inflammatory compounds (C)] or a pharmaceutically acceptable salt, pharmaceutically acceptable solvate, isomer or mixture thereof,

Formula (Ia)

where

- each R ¹ is independently selected from OH, NH ₂ or NH(R ^1b ), where R 1b is a C 1-10 cycloalkyl group or C 1-10 cycloalkenyl which is optionally substituted with halogen, hydroxyl or alkoxy, more preferably R 1 was -NH(R ^1b ), where R ^1b has the following formula:

- each R ³ is independently selected from the group consisting of H, -OH and -OR ^w , where R ^w represents D-glucose;

- each of R ² and R ⁴ , independently of each other and in each case of their presence, is selected from H or -OH;

- each R ⁵ is independently selected from the group consisting of H, -OH and -CH ₃ ;

- Lipid is selected from the following fragments having formula L ₁ or formula L ₂ :

,

,

_Formula L1

Formula _L2

More preferably, the anti-inflammatory compound (C) is a mixture of class I anti-inflammatory compounds (C) as detailed above.

Non-limiting examples of commercially available anti-inflammatory compounds (C) suitable for use in the invention include flavomycin ^® .

The commercially available ^flavomycin® is also well known by its synonymous name bambermycin.

Bambermycin or Flavomycin ^® is a known complex of compounds mainly consisting of moenomycin A and C, in particular, the main compound is moenomycin A having the formula (Ib) described in detail below, and minor compounds are denoted as A ₁₂ , C ₁ , C ₃ and _C4 . Moenomycin A and minor compounds, referred to as A ₁₂ , C ₁ , C ₃ and C4, are compounds of formula (Ia) described above in detail.

In moenomycin A having the formula (Ib) detailed below and in the minor compounds A ₁₂ , C ₁ , C ₃ and C _{4 ,} Lipid has the formula L ₂ , R ² is H, R ¹ is -NH(R ^1b ), where R ^1b has the following formula:

In moenomycin A, R ³ is OR ^w , where R ^w is D-glucose, R ⁴ is OH, and R ⁵ is CH ₃ . In moenomycin A ₁₂ , R ³ is OR ^w , where R ^w is D-glucose, R ⁴ is H, and R ⁵ is OH.

In moenomycin C ₁ , R ³ is H, R ⁴ is H, and R ⁵ is OH. In moenomycin C ₃ , R ³ is H, R ⁴ is OH, and R ⁵ is CH ₃ . In moenomycin C ₄ , R ³ is OH, R ⁴ is OH and R ⁵ is CH ₃ .

Bambermycin or flavomycin® can be obtained from Streptomyces bambergiensis and Streptomyces ghanaensis, Streptomyces ederensis, Streptomyces geysiriensis and related strains, or biosynthesized to form the complex of compounds described above. Since the complex is not subjected to thorough purification, flavomycin® also contains the original components used in the biosynthesis and various intermediate compounds of the biosynthesis.

Bambermycin is also known as flavophospholipol. Bambermycin, flavophospholipol and flavomycin® are used interchangeably in the invention.

A preferred class I anti-inflammatory compound (C) is in particular moenomycin A having the formula (Ib), or a pharmaceutically acceptable salt, a pharmaceutically acceptable solvate, an isomer or a mixture thereof:

Formula (IB)

According to an alternative embodiment of the present invention, the anti-inflammatory compounds (C) are compounds of formula (IIa) [hereinafter referred to in the invention as class II anti-inflammatory compounds (C)] or a pharmaceutically acceptable salt, a pharmaceutically acceptable solvate, an isomer or a mixture thereof:

Formula (IIa)

where

- R ³ represents H or -OH;

- each of R ² and R ⁴ , independently of each other and in each case of their presence, is selected from H or -OH;

- each of R ^6a and R ^6a' , independently of each other and in each case of their presence, is selected from H or -OH, preferably R ^6a is H and R ^6a' is -OH;

- R ⁵ represents H, -OH or CH ₃ ;

- R ¹³ represents -OH or -NHC(=O)CH ₃ ;

- Lipid is selected from the following fragments having formula L ₃ , formula L ₄ or formula L ₅ :

According to an alternative embodiment of the present invention, the anti-inflammatory compounds (C) are compounds of formula (IIIa) [hereinafter referred to in the invention as class III anti-inflammatory compounds (C)] or a pharmaceutically acceptable salt thereof, a pharmaceutically acceptable solvate, an isomer mixture thereof:

Formula (IIIa)

where

- each of R ⁴ and R ⁵ , independently of each other and in each case of their presence, is selected from the group consisting of H, -CH ₃ and -OH;

- R ₇ is independently selected from H or -C(=O)NH ₂ , preferably R ₇ is C(=O)NH ₂ ;

- R ¹⁰ is independently selected from -C(=O)NH ₂ or -C(=O)OH, preferably R ¹⁰ is C(=O)NH ₂ ;

- R ¹³ is independently selected from the group consisting of -OH or -NHC(=O)CH ₃ , preferably R ¹³ is --NHC(=O)CH ₃

- Lipid is selected from the following fragments having formula L ₆ , formula L ₇ or formula L ₈ :

From the point of view of therapeutic use, the salts of the above-described anti-inflammatory compound (C) of the present invention are salts in which the counterion is pharmaceutically acceptable, and the salts of which may be referred to as pharmaceutically acceptable acid addition and base addition salts. However, salts of acids and bases which are not pharmaceutically acceptable may also find use, for example, in the preparation or purification of a pharmaceutically acceptable compound. All salts, whether or not they are pharmaceutically acceptable, are within the scope of the present invention.

The pharmaceutically acceptable acid and base addition salts are intended to include the active non-toxic acid and base addition salt forms which are capable of forming the anti-inflammatory compounds (C) of the present invention described in detail above. Pharmaceutically acceptable acid addition salts can be easily obtained by treating the base form with such an appropriate acid in anionic form. Suitable anions include e.g. , hexylresorcinate, hydrabamine, hydrobromide, hydrochloride, hydroxynaphthoate, iodide, isethionate, lactate, lactobionate, malate, maleate, mandelate, mesylate, methyl bromide, methyl nitrate, methyl sulfate, mucat, napsilate, nitrate, pamoate (embonate), pantothenate, phosphate/diphosphate, polygalacturonate, salicylate, stearate, bumacetate, succinate, sulfate, tannate, tartrate, theoclate, triethiodide and other similar anions. The selected counterion can be introduced using ion exchange resins. Conversely, said salt forms may be converted by treatment with an appropriate base to the free base form.

The anti-inflammatory compounds (C) mentioned in the invention containing an acidic proton can also be converted into their non-toxic metal or amine addition salt forms by treatment with appropriate organic and inorganic bases in cationic form. Suitable base salts include salts formed with organic cations such as benzathine, chlorprocaine, choline, diethanolamine, ethylenediamine, megluumine, procaine and other similar organic cations; and salts formed by metal cations such as aluminum, calcium, lithium, magnesium, potassium, sodium, zinc and the like. Conversely, said salt forms can be converted by treatment with an appropriate acid to the free form.

As used above, the term "addition salt" also includes solvates which are capable of forming the anti-inflammatory compound (C) specified in the invention, as well as salts thereof. Such solvates are, for example, hydrates, alcoholates and the like.

It is well known that inflammation-related diseases involve a complex "system" of cytokine networks in the pathological process, as a result of which the functioning of the immune system is finely balanced by the action of pro-inflammatory and anti-inflammatory mediators or cytokines.

As detailed above, said anti-inflammatory compound (C) of the present invention acts on inflammatory mechanisms in mammals. Preferably, the anti-inflammatory compound (C) of the present invention described in detail above acts on inflammatory mechanisms in non-human mammals. The non-human mammal may be a pig; a ruminant such as, in particular, cattle, horses; camel, sheep, goat, cat, dog; or a rodent such as, in particular, a mouse, a rabbit or a rat, preferably a pig.

The Applicant was surprised to find that the anti-inflammatory compound (C) of the present invention described in detail above can affect the expression of at least one cytokine by suppressing or inhibiting the expression of at least one cytokine, wherein said cytokine is selected from the group consisting of pro-inflammatory cytokines, anti-inflammatory cytokines and chemokines.

In the context of the present invention, it is intended that the expressions "at least one cytokine" mean one or more than one cytokine. In other words, the anti-inflammatory compound (C) of the present invention can suppress or inhibit the expression of one, two, three or more cytokines, and thereby achieve higher therapeutic efficacy with fewer side effects.

In addition, the Applicant has found that by suppressing or inhibiting the expression of at least one cytokine by using the anti-inflammatory compound (C) of the present invention, there is a significant reduction in the infiltration of macrophages, B cells and T cells. This indicates that the anti-inflammatory compound (C) of the present invention has the property of suppressing or inhibiting inflammation.

Examples of diseases mediated by cytokines (C) and which are treatable by using the anti-inflammatory compound (C) of the present invention specifically include inflammatory diseases and inflammation-related diseases.

Non-limiting examples of inflammatory diseases specifically include arthritis, rheumatoid arthritis, inflammatory bowel disease, psoriasis, multiple sclerosis, systemic lupus erythematosus (SLE), pancreatitis, scleroderma, type I diabetes, inflammatory bowel disease, allergic conjunctivitis, glomerulonephritis, Sjögren's syndrome, uveitis, dermatitis, ankylosing spondylitis and fibromyalgia.

It should be understood that the term "inflammatory bowel disease" refers to diseases such as acute gastritis, chronic gastritis, development and recurrence of gastric ulcers, Crohn's disease, inflammatory bowel disease, irritable bowel syndrome and ulcerative colitis.

Non-limiting examples of inflammation-related diseases include, but are not limited to, neurodegenerative disorder, congestive heart failure, stroke, aortic valve stenosis, renal failure, allergy, fibrosis, atherosclerosis, metabolic disease, cardiovascular disease, chemotherapy/radiotherapy complication , liver disease, gastrointestinal disorder, ophthalmic disease, diabetic retinopathy, pulmonary disease, and leprosy.

Proinflammatory cytokines specifically include IL-1α, IL-1β, IL-2, IL-3, IL-6, IL-7, IL-9, IL-12, IL-17, IL-18, IL-23 , TNF-α, LT, LIF, oncostatin and IFN-α, IFN-β and IFN-γ.

Anti-inflammatory cytokines specifically include IL-4, IL-10, IL-11, W-13 and TGF-β.

Chemokines specifically include IL-8, Gro-α, MIP-1, MCP-1, ENA-78, and RANTES (a chemokine expressed and secreted by T cells upon activation).

Unless otherwise noted or indicated, the designated cytokines detailed above include all possible homologous forms that can be suppressed by the indicated anti-inflammatory compounds (C). Homology is defined as the mutual similarity between biological structures, proteins, or DNA sequences that stems from a common ancestor. For example, the term "IL-8" refers to IL-8 and all IL-8 homologues, such as IL-8 _Kc , IL-8 _MIP, IL-8 _LIX and other such cytokines in particular.

PHARMACEUTICAL COMPOSITIONS

In addition, the present invention relates to a pharmaceutical composition comprising a therapeutically effective amount of an anti-inflammatory compound (C) as defined above and as defined in any one of the embodiments presented in the invention, in particular, an anti-inflammatory compound (C) of any of the classes (I) defined above - (III), and a pharmaceutically acceptable carrier.

Hereinafter, the expression "anti-inflammatory compound (C)" means, in relation to the purposes of the present invention, both the plural and the singular, that is, it means that the composition according to the invention may include one or more than one anti-inflammatory compound (C) "

In yet another aspect, this invention relates to a method of preparing a pharmaceutical composition as detailed above, which comprises intimately mixing a pharmaceutically acceptable carrier with a therapeutically effective amount of an anti-inflammatory compound (C) as defined above and as defined in any one of the embodiments of the invention, in particular , anti-inflammatory compound (C) of any of the above-defined classes (I) - (III).

Therefore, said anti-inflammatory compounds (C) of the present invention, in particular, anti-inflammatory compounds (C) of any of the classes (I) to (III) defined above, can be prepared to be administered in various dosage forms. As suitable compositions there may be mentioned all compositions commonly used for the systemic administration of drugs. For the preparation of pharmaceutical compositions of this invention, an effective amount of a particular compound, optionally in the form of an addition salt or a metal complex, as the active ingredient is combined by intimate mixing with a pharmaceutically acceptable carrier which may take a wide variety of forms, depending on the formulation desired. for an introduction. Desirably, these pharmaceutical compositions are in unit dosage form, in particular for oral, rectal, transdermal or parenteral administration. For example, when preparing compositions in a dosage form for oral administration, conventional pharmaceutical media can be used, such as, for example, water, glycols, oils, alcohols and the like, in the case of oral liquid preparations, such as suspensions, syrups, elixirs, emulsions and solutions; or solid carriers such as starches, sugars, kaolin, lubricants, binders, disintegrants and the like in the case of powders, pills, capsules and tablets.

Because of their ease of administration, tablets and capsules are the most convenient single dose oral dosage forms, in which case, of course, solid pharmaceutical carriers are used. For parenteral compositions, the carrier will usually comprise sterile water, at least in large part, although other ingredients may be included, for example, to improve solubility. Injectable solutions may be prepared in which, for example, the carrier comprises saline, glucose solution, or a mixture of saline and glucose solution. Injectable suspensions may also be prepared in which case appropriate liquid carriers, suspending agents and the like may be used. They also include solid form preparations which are intended to be converted, shortly before use, to liquid form preparations. In compositions used for transdermal administration, the carrier optionally comprises a drug penetration enhancer and/or a suitable wetting agent, optionally combined with suitable additives of any nature in minor amounts that do not cause significant adverse effects on the skin.

The anti-inflammatory compound (C) as defined above and as defined in any one of the embodiments of the invention, in particular the anti-inflammatory compounds (C) of any of the classes (I) to (III) defined above, may also be administered by oral inhalation or insufflation using methods and preparations used in pharmaceuticals for administration in this way. Thus, in general, the anti-inflammatory compound (C) specified in the invention, in particular, anti-inflammatory compounds (C) of any of the classes (I) to (III) defined above, can be administered to the lungs in the form of a solution, suspension or dry powder, and the solution is the preferred form. Any system designed to deliver solutions, suspensions, or dry powders by oral inhalation or insufflation is suitable for administering the compounds of the present invention.

It is particularly advantageous to formulate the above-mentioned pharmaceutical compositions in single dose dosage form from the viewpoint of ease of administration and uniformity of dosing. As used herein, a single dose dosage form refers to physically discrete units suitable as unit doses, each unit containing a predetermined amount of the active ingredient calculated to produce the desired therapeutic effect, together with the required pharmaceutical carrier. Examples of such unit dose dosage forms are tablets (including scored or coated tablets), capsules, pills, suppositories, powder sachets, implant capsules, injectable solutions or suspensions, and the like, and divided multiples thereof.

It should be understood that the term "therapeutically effective amount" generally refers to the amount of an active compound or component or drug that allows a biological or drug response to be achieved in a tissue, system, animal or human being sought, in light of the present invention, by the researcher. , veterinarian, physician or other clinician, which includes alleviating the symptoms of the disease being treated.

The present invention contemplates that a therapeutically effective amount of an anti-inflammatory compound (C) as defined above, in particular an anti-inflammatory compound (C) of any of the classes (I) to (III) defined above, refers to an amount sufficient to prevent, stabilize or alleviate disease mediated by at least one cytokine selected from the group consisting of pro-inflammatory cytokines, anti-inflammatory cytokines and chemokines described above, in particular at least one cytokine selected from the group consisting of IL-8, Gro- α, MIP-1, MCP-1, ENA-78 and RANTES, IL-1α, IL-1β, IL-2, IL-3, IL-6, IL-7, IL-9, IL-12, IL- 17, IL-18, IL-23, TNF-α, LT, LIF, oncostatin, IFN-α, IFN-β, IFN-γ, IL-4, IL-10, IL-11, W-13 and TGF- β, more preferably at least one cytokine is selected from the group consisting of IL-8, IL-4, IL-6, IL-10, TNF-α, TNF-β, IL-17, IL-23 and IFN-γ, even more preferably S, at least one of these cytokines was selected from the group consisting of IL-8, IL-1β and IFN-γ.

USE OF ANTI-INFLAMMATORY COMPOUNDS (C) AND PHARMACEUTICAL COMPOSITIONS AS MEDICINES AND IN THE TREATMENT OF DISEASES

The present invention relates to an anti-inflammatory compound (C) as defined above, in particular an anti-inflammatory compound (C) of any of the classes (I) to (III) defined above, which is used as a medicine.

The present invention also relates to a pharmaceutical composition comprising a therapeutically effective amount of an anti-inflammatory compound (C) as defined above and as defined in any one of the embodiments presented in the invention, in particular, an anti-inflammatory compound (C) of any of the classes (I) defined above - ( III), and a pharmaceutically acceptable carrier that is used as a drug.

The present invention relates to an anti-inflammatory compound (C) as defined above, in particular an anti-inflammatory compound (C) of any of the classes (I) to (III) defined above, which is used in the treatment of inflammatory diseases selected from the group of arthritis, rheumatoid arthritis, inflammatory bowel disease, psoriasis, multiple sclerosis, systemic lupus erythematosus (SLE), pancreatitis, scleroderma, type I diabetes, inflammatory bowel disease, allergic conjunctivitis, glomerulonephritis, Sjögren's syndrome, uveitis, dermatitis, ankylosing spondylitis and fibromyalgia, preferably for use in the treatment of inflammatory diseases of the gastrointestinal tract, such as acute gastritis, chronic gastritis, development and recurrence of gastric ulcers, Crohn's disease, inflammatory bowel disease, irritable bowel syndrome and ulcerative colitis, more preferably for use in the treatment of acute gastritis, chronic gastritis, development and recurrence of stomach ulcers.

The present invention relates to an anti-inflammatory compound (C) as defined above, in particular an anti-inflammatory compound (C) of any of the classes (I) to (III) defined above, which is used in the treatment of inflammation-related diseases selected from the group of neurodegenerative disorder, congestive heart failure, stroke, aortic valve stenosis, renal failure, allergies, fibrosis, atherosclerosis, metabolic disease, cardiovascular disease, chemotherapy/radiation therapy complications, liver disease, gastrointestinal disorders, eye disease, diabetic retinopathy, lung disease and leprosy.

Therefore, the anti-inflammatory compound (C) as defined above, in particular the anti-inflammatory compound (C) of any of the classes (I) to (III) defined above, and the pharmaceutical composition as defined above can be used in the manufacture of a medicament used for the treatment, prevention or counteracting a disorder or disease associated with cytokine expression, including inflammatory diseases as defined above and inflammation-associated diseases as defined above.

The present invention relates to an anti-inflammatory compound (C) as defined above, in particular an anti-inflammatory compound (C) of any of the classes (I) to (III) defined above, which is used in the treatment of a disease mediated by at least one cytokine.

At least one cytokine is preferably selected from the group consisting of pro-inflammatory cytokines, anti-inflammatory cytokines and chemokines as detailed above, preferably at least one cytokine is selected from the group consisting of IL-8, Gro-α, MIP- 1, MCP-1, ENA-78 and RANTES, IL-1α, IL-1β, IL-2, IL-3, IL-6, IL-7, IL-9, IL-12, IL-17, IL- 18, IL-23, TNF-α, LT, LIF, oncostatin, IFN-α, IFN-β, IFN-γ, IL-4, IL-10, IL-11, W-13 and TGF-β, more preferably that at least one cytokine is selected from the group consisting of IL-8, IL-4, IL-6, IL-10, TNF-α, TNF-β, IL-17, IL-23 and IFN-γ , even more preferably, at least one of these cytokines is selected from the group consisting of IL-8, IL-1β and IFN-γ.

It is well known to those skilled in the art of medicine that the exact dosage and frequency of administration depend on the specific anti-inflammatory compound (C) used, the particular condition being treated, the severity of the condition being treated, age, body weight, sex, degree of disorder, and general physical condition of a particular patient, as well as from another drug that the patient may be taking. In addition, it is obvious that the specified effective daily amount can be reduced or increased depending on the response of the subject being treated and/or depending on the judgment of the attending physician who prescribed the use of the compounds of the present invention.

As mentioned above, the above-described anti-inflammatory compound (C) of the present invention can act on inflammatory mechanisms in animals.

In the case of administration to animals, an anti-inflammatory compound (C) as defined above, in particular an anti-inflammatory compound (C) of any of the classes (I) to (III) defined above, and a pharmaceutical composition as defined above comprising at least one of said anti-inflammatory compounds (C) as an active ingredient, can be added to animal feed or drinking water, can be administered by spraying as a probiotic to a nursing female, can be administered in the form of a sustained release bolus or in the form of a lick stone.

An effective amount of an anti-inflammatory compound (C) as defined above, in particular an anti-inflammatory compound (C) of any of (I) to (III) as defined above, which specifically suppresses or inhibits the expression of at least one cytokine detailed above, for the treatment of animals is preferably at least 1 ppm, preferably at least 2 ppm, more preferably at least 3 ppm, based on the total weight of the animal feed. On the other hand, the effective amount of the anti-inflammatory compound (C) defined above, in particular the anti-inflammatory compound (C) of any of the above (I) to (III), composition (C) is preferably less than 130 ppm, preferably less than than 70 ppm, more preferably less than 35 ppm, based on the total weight of the animal feed, where the animal in this case is a rodent, in particular a mouse. It should be borne in mind that, in relation to the purposes of the present invention, the value of the upper limit is not critical, and the effective amount depends on the particular species of animal being treated. The invention further relates to an animal feed comprising an anti-inflammatory compound (C) as defined above, in particular an anti-inflammatory compound (C) of any of the classes (I) to (III) defined above, a pharmaceutical composition comprising said anti-inflammatory compound (C ) as the active ingredient.

METHODS OF TREATMENT OF INFLAMMATORY DISEASES USING ANTI-INFLAMMATORY COMPOUNDS (C)

The present invention relates to an in vivo method for suppressing or inhibiting inflammatory responses by using at least one anti-inflammatory compound (C) as defined above, in particular an anti-inflammatory compound (C) of any of the classes (I) to (III) defined above. and defined in any one of the embodiments of the invention.

In addition, the present invention relates to a method for suppressing or inhibiting the expression of at least one cytokine in a mammal, which method includes administering to a mammal in need thereof an amount of an anti-inflammatory compound (C) as defined above and as defined in any one of the following. in the invention embodiments, sufficient to inhibit or suppress the expression of at least one cytokine, where the cytokine is selected from the group consisting of IL-8, Gro-α, MIP-1, MCP-1, ENA-78 and RANTES, IL -1α, IL-1β, IL-2, IL-3, IL-6, IL-7, IL-9, IL-12, IL-17, IL-18, IL-23, TNF-α, LT, LIF , oncostatin, IFN-α, IFN-β, IFN-γ, IL-4, IL-10, IL-11, W-13 and TGF-β, more preferably at least one cytokine is selected from the group, consisting of IL-8, IL-4, IL-6, IL-10, TNF-α, TNF-β, IL-17, IL-23 and IFN-γ, even more preferably at least one of these cytokines were selected from the group consisting of IL-8, IL-1β and IFN-γ.

In addition, the present invention relates to a method of treating inflammation-related diseases associated with cytokine expression levels in a mammal, which method includes administering to a mammal in need thereof an amount of an anti-inflammatory compound (C) as defined above and as defined in any one of those presented in embodiments of the invention effective for treating said inflammation-related disease.

The following examples are only illustrations of non-limiting embodiments of the present invention.

EXAMPLES

MATERIALS

a) Setting up the experiment

Experiments were carried out in a mouse model to evaluate the effect of feed additives with flavomycin® as an anti-inflammatory compound (C).

For in vivo experiments, the BALB/c mouse strain was chosen due to the fact that this strain of mice has a more pronounced pathology of the stomach compared to other strains of mice. Mice were divided into 4 groups of 8 animals each. The 4 groups have the following characteristics:

1) infected + standard diet: comparative example 1

Mice of the "comparative example 1" group were inoculated intragastrically with 108 viable Helicobacter suis (H. suis) HS1 strains twice for 48 hours (day 8 and day 9). On day 17, mice were given ad libitum access to the control diet (standard diet without flavomycin®). After 73 days, mice were sacrificed by dislocation of the cervical vertebrae under deep anesthesia with isoflurane. A strip of gastric tissue was taken to perform the experimental procedures described below.

2) Infected + Flavomycin® Supplemented Diet (Amount of Flavomycin® <100 ppm based on total diet provided): Example 1

Mice of the "Example 1" group were inoculated twice intragastrically with 108 viable Helicobacter Suis (H. suis) HS1 strain bacteria for 48 hours (Day 8 and Day 9). On day 17, mice were given ad libitum access to the flavomycin® supplemented diet (flavomycin® <100 ppm based on total diet provided). After 73 days, mice were sacrificed by dislocation of the cervical vertebrae under deep anesthesia with isoflurane. A strip of gastric tissue was taken to perform the experimental procedures described below.

3) non-infected + standard diet: comparative example 2

The mice of the "comparative example 2" group were not subjected to intragastric inoculation and were given ad libitum access to the control chow, a standard diet without flavomycin®. After 73 days, mice were sacrificed by dislocation of the cervical vertebrae under deep anesthesia with isoflurane. A strip of gastric tissue was taken to perform the experimental procedures described below.

4) Uninfected + Flavomycin® Supplemented Diet (Amount of Flavomycin® <100 ppm based on total feed provided): Example 2

Mice of the "Example 2" group were not subjected to intragastric inoculation and were given ad libitum access to the diet supplemented with flavomycin® (amount of flavomycin® <100 ppm based on the total amount of diet provided). After 73 days, mice were sacrificed by dislocation of the cervical vertebrae under deep anesthesia with isoflurane. A strip of gastric tissue was taken to perform the experimental procedures described below.

b) Verification of the experimental setup - quantification of H. suis

The presence of H. suis infection was determined by real-time polymerase chain reaction (qRT-PCR). Experiments were carried out as detailed below. Table 1 shows that the "Comparative Example 1" and "Example 1" groups were infected with H. suis bacteria. The "Comparative Example 2" and "Example 2" groups were not infected as shown in Table 1 because H. suis bacteria were not detected in these animals.

Table 1. Determination of the presence of H. suis infection by qRT-PCR expressed as logarithmic ratios of H. suis per mg of tissue (absolute nucleic acid quantitation (amount of bacterial DNA) per given amount of sample (mg of tissue).

EXAMPLE Log H.suis/mg tissue Comparative Example 1 7,880 Example 1 36,900 Comparative Example 2 0 Example 2 0

METHODS

Real-time polymerase chain reaction (qRT-PCR) analysis

1) Quantification of H.suis

J.L. et al. (2004). Присутствие ДНК H.suis в экстрактах определяли с использованием специфичного для H. suis метода qRT-PCR с абсолютной количественной оценкой, основанной на гене ureA (Blaecher C. et al. (2013)). Сначала генерировали стандарт путем амплификации части кластера генов ureAB (1236 п.о.) из штамма H.suis HS5 с использованием праймеров U430F и U1735R (

J.L. et al. (2004)). Вкратце, реакции амплификации PCR включали 1 × реакционный буфер [67 мМ Tris/HCl, 16 мМ (NH₄)₂SO₄, 0,45% Triton Х-100, 0,2% желатин], одну единицу ДНК-полимеразы Taq (Biotech International), 200 мкМ каждого дезоксинуклеотидтрифосфата, 2 мМ MgCl₂, 10 пмоль каждого олигонуклеотидного праймера и 1 мкл разведенной ДНК (обычно разведение исходного образца в соотношении 1:10, содержащее приблизительно 20-100 нг/мкл) в конечном объеме 50 мкл. Условиями циклирования были начальная денатурация при 94°С в течение 3 минут, затем 35 циклов при 94°С в течение 10 секунд, 52°С в течение 30 секунд и 72°С в течение 1,5 минут с последующей конечной стадией элонгации при 72°С в течение 5 минут. Все реакции проводили с использованием термоциклера Perkin Elmer PE2400. Продукты PCR разделяли на мини-гелях с агарозой в буфере TAE (40 мМ Tris/ацетат, 1 мМ ЭДТА) и фотографировали при УФ-трансиллюминации после окрашивания бромидом этидия. Продукты PCR очищали перед секвенированием с помощью системы очистки Wizard PCR Preps DNA Purification System (Promega) или фильтров Centricon-100 (Amicon). Затем амплифицированную ДНК непосредственно секвенировали с использованием набора для секвенирования циклов ABI PRISM Ready Reaction DyeDeoxy Terminator Cycle Sequencing Kit (PE Applied Biosystems) и системы GeneAmp® PCR System 2400 (Perkin Elmer) в соответствии с протоколами фирм-производителей. Продукты секвенирования разделяли на секвенаторах model 377 DNA Sequencer и анализировали с использованием программ, содержащихся в прилагаемом пакете (PE Applied Biosystems). Во всех случаях, обе цепи ДНК секвенировали с помощью непрерывных перекрытий. После получения стандарта, были сделаны 10-кратные разведения, начиная с 108 ампликонов PCR, для каждого 9 мкл реакционной смеси. Один микролитр экстрагированной ДНК-матрицы добавляли к 9 мкл реакционной смеси, состоящей из 0,25 мкл обоих праймеров, расположенных внутри фрагмента 1236 п.о. (для получения продукта PCR 150 п.о.), 3,5 мкл воды хроматографической чистоты и 5 мкл SensiMix™ SYBR No-ROX (Bioline Reagents Ltd, London, UK). Смысловой праймер представлял собой BF_HsuisF1: 5'-AAA ACA MAG GCG ATC GCC CTG TA-3'. Антисмысловой праймер представлял собой BF_Hsuis R1: 5'-TTT CTT CGC CAG GTT CAA AGC G-3'. Температура отжига составила 62°С. Оба стандарта и образцы были подвергнуты двум параллельным анализам на системе CFX96™ RT-PCR System с термоциклером C1000 Thermal Cycler (Bio-Rad, Hercules, CA, USA). Результаты выражены в единицах абсолютного количества нуклеиновой кислоты (количества бактериальной ДНК) на данное количество образца (мг ткани).Quantification of the formation of H. suis colonies in the stomach was carried out in accordance with the publications of Blaecher C. et al. (2013) and

JL et al. (2004). The presence of H. suis DNA in the extracts was determined using the H. suis-specific qRT-PCR method with absolute quantification based on the ureA gene (Blaecher C. et al. (2013)). First, a standard was generated by amplifying a portion of the ureAB gene cluster (1236 bp) from the H. suis HS5 strain using primers U430F and U1735R (

JL et al. (2004)). Briefly, PCR amplification reactions included 1× reaction buffer [67 mM Tris/HCl, 16 mM (NH ₄ ) ₂ SO ₄ , 0.45% Triton X-100, 0.2% gelatin], one unit of Taq DNA polymerase ( Biotech International), 200 μM of each deoxynucleotide triphosphate, 2 mM MgCl ₂ , 10 pmol of each oligonucleotide primer, and 1 μl of diluted DNA (typically a 1:10 dilution of the original sample containing approximately 20-100 ng/μl) in a final volume of 50 μl. Cycling conditions were initial denaturation at 94°C for 3 minutes followed by 35 cycles at 94°C for 10 seconds, 52°C for 30 seconds and 72°C for 1.5 minutes followed by a final elongation step at 72 °C for 5 minutes. All reactions were performed using a Perkin Elmer PE2400 thermal cycler. PCR products were resolved on agarose mini-gels in TAE buffer (40 mM Tris/acetate, 1 mM EDTA) and photographed under UV transillumination after staining with ethidium bromide. PCR products were purified prior to sequencing using the Wizard PCR Preps DNA Purification System (Promega) or Centricon-100 filters (Amicon). The amplified DNA was then directly sequenced using the ABI PRISM Ready Reaction DyeDeoxy Terminator Cycle Sequencing Kit (PE Applied Biosystems) and GeneAmp® PCR System 2400 (Perkin Elmer) according to the manufacturer's protocols. The sequencing products were separated on model 377 DNA Sequencers and analyzed using the programs contained in the attached package (PE Applied Biosystems). In all cases, both strands of DNA were sequenced using continuous overlaps. After obtaining the standard, 10-fold dilutions were made, starting with 108 PCR amplicons, for each 9 μl of the reaction mixture. One microliter of the extracted DNA template was added to 9 μl of the reaction mixture consisting of 0.25 μl of both primers located within the 1236 bp fragment. (to obtain a 150 bp PCR product), 3.5 µl chromatographic grade water, and 5 µl SensiMix™ SYBR No-ROX (Bioline Reagents Ltd, London, UK). The sense primer was BF_HsuisF1: 5'-AAA ACA MAG GCG ATC GCC CTG TA-3'. The antisense primer was BF_Hsuis R1: 5'-TTT CTT CGC CAG GTT CAA AGC G-3'. The annealing temperature was 62°C. Both standards and samples were run in duplicate on the CFX96™ RT-PCR System with C1000 Thermal Cycler (Bio-Rad, Hercules, CA, USA). The results are expressed in terms of the absolute amount of nucleic acid (amount of bacterial DNA) per given amount of sample (mg of tissue).

2). Cytokine expression levels

Cytokine expression levels were assessed by qRT-PCR according to Flahou B. et al. (2012) and Liu C. et al. (2016). RNA was extracted using the RNeasy Mini Kit® (Qiagen, Hilden, Germany) according to the manufacturer's instructions. The resulting RNA concentrations were measured using a NanoDrop® spectrophotometer (Isogen Life Science, Utrecht, The Netherlands), after which the concentration of all samples taken from all mice belonging to the 4 groups defined above in the experimental setup section was adjusted to 1 µg/µl , then cDNA was synthesized using the iScript™ cDNA Synthesis Kit (Bio-Rad, California, USA). Then, expression analysis was performed for genes encoding host factors involved in inflammation. The constitutive genes PPIa, H2afz and HPRT were included as reference genes. All primer sequences used are shown in Table 2. mRNA expression levels of the reference and target genes were quantified using the reverse transcriptase polymerase chain reaction (RT-PCR) method described previously (Flahou B. et al. (2012)). Matrix-controlled reaction mixtures were used and all of these samples were subjected to two replicate analyses. Cycle threshold (Ct) values were first normalized to the geometric mean Ct of the reference genes. The multiplicity of changes was calculated by the ΔΔCT method (72) from the average Ct values in mice with a negative result for the presence of H. suis. Finally, for each target gene, the results were expressed as fold changes in mRNA expression levels in H. suis positive mice relative to mRNA expression levels in H. suis negative mice. The results are shown in tables 3 and 4 below.

Table 2. Primer sequences used in the qRT-PCR method

Sequence (5'-3') primer CTGACCTAGAGAAGACACAT IFN-γ - direct GGTCAGTGAAGTAAAGGTAC IFN-γ - reverse CACCTCACAAGCAGAGCACAAG IL-1β - direct GCATTAGAAAACAGTCCAGCCCATAC IL-1β - reverse TGCCTGAAGACCCTGCCAAGG IL-8 _MIP - straight GTTAGCCTTGCCTTTGTTCAG IL8 _MIP - Reverse Constitutive genes GGTATCACCCCTCGTCACTT H2afz - straight TCAGCGATTTGTGGATGTGT H2afz - Reverse CAGGCCAGACTTTGTTGGAT HPRT - straight TTGCGCTCATCTTAGGCTTT HPRT - Reverse AGCATACAGGTCCTGGCATC PPIa - straight TTCACCTTCCCAAAGACCAC PPIa - reverse

Immunohistochemical studies - determination of immunohistochemistry of T cells, B cells, parietal cells, necrotic cells and macrophage infiltration

Immunohistochemical studies were performed in accordance with the publication of Flahou B. et al. (2010). Histological sections successively 5 μm thick were made from paraffin-embedded tissues. After rehydration and deparaffinization, heat-induced antigen unmasking was performed in citrate buffer (pH 6.0) using a microwave oven. Slides were incubated with 3% H ₂ O ₂ in methanol (5 minutes) and 30% goat serum (30 minutes) to block endogenous peroxidase activity and non-specific reactions, respectively. Differentiation between T and B lymphocytes was performed by staining for CD3 and CD20 antigens. CD3 antigens were detected using a polyclonal rabbit anti-CD3 antibody (1/100; DakoCytomation). CD20 antigens were detected using a polyclonal anti-CD20 rabbit antibody (1/100; Thermo Scientific, Fremont, USA), respectively. Incubation with primary antibodies against CD3 and CD20 was followed by incubation with biotinylated goat anti-rabbit IgG (1/500; DakoCytomation). After washing, sections were incubated with streptavidin-biotin-HRP complex (DakoCytomation) and stained with diaminobenzidine tetrahydrochloride (DAB) and H ₂ O ₂ . Primary antibody against cell surface marker F4/80 (1/50; Santa Cruz Biotechnology, Inc., Santa Cruz, USA) was used to isolate mature macrophages. Detection was performed using a rat ABC staining system (Santa Cruz Biotechnology, Inc.). Apoptotic cells were identified by immunohistochemical staining using a rabbit polyclonal antibody against active caspase-3 and an anti-rabbit HRP-AEC cell and tissue staining kit (R&D Systems, Minneapolis, USA). Parietal cells were identified by immunohistochemical staining for hydrogen potassium adenosine triphosphatase using a mouse monoclonal antibody (1/200; Abcam Ltd, Cambridge, UK) and a biotinylated goat anti-mouse IgG antibody (1/200; DakoCytomation). Apoptotic cells were identified by immunohistochemical staining using a rabbit polyclonal antibody against active caspase-3 and an anti-rabbit HRP-AEC cell and tissue staining kit (R&D Systems, Minneapolis, USA). Parietal cells were identified by immunohistochemical staining for hydrogen potassium adenosine triphosphatase using a mouse monoclonal antibody (1/200; Abcam Ltd, Cambridge, UK) and a biotinylated goat anti-mouse IgG antibody (1/200; DakoCytomation). Finally, positive cells were counted in five randomly selected high magnification microscope fields (magnification: x400). The average number of positive cells was then determined for each mouse. All results are presented in tables 5, 6, 7 and 8.

RESULTS

Experiment 1

Measurement of cytokine expression levels in H. suis infected animals using the qRT-PCR method described in detail above.

Table 3. Results of a qRT-PCR experiment to determine the effects of flavomycin supplementation ^and on cytokine levels in H. Suis infected animals, expressed as fold change in mRNA expression levels.

Expression (fold change) Cytokine Comparative Example 1 Example 1 IL-1β 0.82 -1.38 IL-8MIP 2.65 1.60 IFN-γ 2.15 -3.88

Table 3 shows the reduction in expression levels of IL-1β, IL-8MIP and IFN-γ in infected animals fed a diet supplemented with ^flavomycin® (Example 1) compared to a control group fed a standard diet (Comparative Example 1).

Experiment 2

Measurement of cytokine expression levels in the absence of H. suis infection using the qRT-PCR method detailed above.

Table 4 Results of a qRT-PCR experiment to determine the effect of flavomycin® supplementation on cytokine levels in uninfected animals, expressed as fold change in mRNA expression levels.

Expression (fold change) Cytokine Comparative example 2 Example 2 IL-1β 1.08 -1.42 IL-8MIP 2.47 2.34 IFN-γ 1.25 1.99

Table 4 shows the decrease in IL-1β expression even in the absence of infection. IL-8MIP expression levels remain stable, with a slight increase in IFN-γ expression.

Experiment 3

Evaluation of the inflammatory response in H. suis infected animals by conducting the immunohistochemical studies described in detail above.

Table 5. Results of immunohistochemical studies to determine the effect of ^flavomycin® supplementation on the number of infiltrating T cells, B cells and macrophages in H. Suis infected animals, expressed as the mean number of positive cells counted.

Inflammation index Comparative Example 1 Example 1 T cells 60.28 21.38 B cells 3.08 0.48 Macrophages 4.75 1.35

Table 5 shows a strong decrease in the number of infiltrating T cells, B cells and macrophages in H. suis infected mice treated with ^flavomycin® (Example 1) compared to Comparative Example 1.

Experiment 4

Evaluation of the inflammatory response in the absence of H. suis infection by conducting the immunohistochemical studies described in detail above.

Table 6. Results of immunohistochemical studies to determine the effect of ^flavomycin® supplementation on the number of infiltrating T-cells, B-cells and macrophages in uninfected animals, expressed as the average number of positive cells counted.

Inflammation score Comparative Example 2 Example 2 T cells 54.00 14.10 B cells 0.85 1.65 Macrophages 1.15 0.18

Table 6 shows the anti-inflammatory effect of ^flavomycin® in the group of non-infected mice fed a supplemented diet (Example 2) compared to Comparative Example 2. In particular, a decrease in the number of T cells and macrophages was observed.

Based on the results of experiments 3 and 4, it is shown that flavomycin® has an anti-inflammatory effect, regardless of the presence of H. suis infection.

Experiment 5

Physiological evaluation of H. suis infected animals by performing the immunohistochemical studies described in detail above.

Table 7. Results of immunohistochemical studies to determine the effect of flavomycin ^® supplementation on the number of necrotic and parietal cells in H. Suis infected animals, expressed as the average number of positive cells counted.

Physiological indicator Comparative Example 1 Example 1 Necrotic cells 0.95 0.50 parietal cells 151.25 213.25

Table 7 shows the reduction in the number of necrotic cells for a group of infected animals fed a diet supplemented with flavomycin ^® (Example 1) compared to comparative example 1. This indicates that animals fed a diet supplemented with flavomycin ^® experience less cellular necrosis than that which could be explained by the suppression of inflammatory responses. As regards the number of parietal cells, a higher number of cells is found, measured in the group of animals treated with a diet supplemented with flavomycin ^® . This can be explained by the fact that the reduction in inflammation resulting from the administration of flavomycin ^® causes less damage to the parietal cells of the stomach.

Experiment 6

Physiological assessment of H. suis non-infected animals by performing the immunohistochemical studies described in detail above.

Table 8. Results of immunohistochemical studies to determine the effect of flavomycin ^® supplementation on the number of necrotic and parietal cells in uninfected animals, expressed as the average number of positive cells counted.

Physiological indicator Comparative Example 1 Example 1 Necrotic cells 2.18 0.60 parietal cells 214.63 211.38

Table 8 shows the reduction in the number of necrotic cells resulting from the reduction in inflammation. The number of parietal cells is approximately the same value as in example 2 and in comparative example 2. This indicates that treatment with flavomycin does not adversely affect the number of parietal cells.

Taking into account all the results obtained, it can be concluded that the administration of flavomycin ^® has an anti-inflammatory effect, since the infiltration of B-cells, T-cells and macrophages, as well as the levels of IL-1β, IL-8MIP and IFN-γ decreased in groups of animals treated with a diet supplemented with flavomycin ^® . Moreover, it has been shown that the anti-inflammatory effect is independent of the presence of H. suis infection.

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Claims (13)

1. The use of a compound hereinafter referred to as an anti-inflammatory compound (C) as an anti-inflammatory agent that acts on the mechanisms of inflammation in mammals, where said anti-inflammatory compound (C) is a compound represented by formula (Ib) or a pharmaceutically acceptable salt thereof, pharmaceutically acceptable hydrate or mixture thereof:

Formula (Ib). 2. The use of an anti-inflammatory compound (C) according to claim 1, wherein said anti-inflammatory compound (C) acts on the mechanisms of inflammation in non-human mammals, in particular the non-human mammal is a pig; a ruminant such as, in particular, cattle, horses; camel sheep goat; cat dog or a rodent, such as, in particular, a mouse, rabbit or rat; preferably a pig. 3. The use of anti-inflammatory compounds (C) according to any one of paragraphs. 1 or 2, wherein said anti-inflammatory compound (C) affects the expression of at least one cytokine by suppressing or inhibiting the expression of at least one cytokine, wherein said cytokine is selected from the group consisting of pro-inflammatory cytokines, in particular IL-1α, IL- 1β, IL-2, IL-3, IL-6, IL-7, IL-9, IL-12, IL-17, IL-18, IL-23, TNF-α, LT, LIF, oncostatin and IFN- α, IFN-β, IFN-γ or mixtures thereof; from anti-inflammatory cytokines, in particular IL-4, IL-10, IL-11, W-13, TGF-β or mixtures thereof; and from chemokines, in particular IL-8, Gro-α, MIP-1, MCP-1, ENA-78, RANTES or mixtures thereof. 4. The use of anti-inflammatory compounds (C) according to any one of paragraphs. 1-3 in the treatment of inflammatory diseases or inflammation-related diseases, wherein said inflammatory diseases or inflammation-related diseases are mediated by at least one cytokine as defined in claim 3. 5. Use of an anti-inflammatory compound (C) according to claim 4, wherein the inflammatory disease is selected from the group consisting of arthritis, rheumatoid arthritis, inflammatory bowel disease, psoriasis, multiple sclerosis, systemic lupus erythematosus (SLE), pancreatitis, scleroderma, type I diabetes , inflammatory disease of the gastrointestinal tract, allergic conjunctivitis, glomerulonephritis, Sjögren's syndrome, uveitis, dermatitis, ankylosing spondylitis and fibromyalgia. 6. The use of the anti-inflammatory compound (C) according to claim 5, wherein the inflammatory disease is selected from the group consisting of inflammatory diseases of the gastrointestinal tract, such as acute gastritis, chronic gastritis, gastric ulcer development and recurrence, Crohn's disease, inflammatory bowel disease, irritable bowel syndrome and ulcerative colitis. 7. Use of the anti-inflammatory compound (C) of claim 4, wherein the inflammation-related disease is selected from the group consisting of neurodegenerative disorder, congestive heart failure, stroke, aortic valve stenosis, renal failure, allergy, fibrosis, atherosclerosis, metabolic disease, cardiac – vascular disease, chemotherapy/radiation therapy complications, liver disease, gastrointestinal disorders, ophthalmic disease, diabetic retinopathy, pulmonary disease and leprosy. 8. Use of the pharmaceutical composition as an anti-inflammatory agent, wherein the pharmaceutical composition comprises a therapeutically effective amount of the anti-inflammatory compound (C) as defined in claim 1 and a pharmaceutically acceptable carrier. 9. The use of a pharmaceutical composition according to claim 8, wherein said pharmaceutical composition is in the form of a single dose dosage form suitable for oral, rectal, transdermal or parenteral administration, in particular in the form of tablets, capsules, pills, suppositories, powder sachets, capsules-implants, injectable solutions or suspensions and in other similar forms, and their divided multiples. 10. In vivo method for suppressing or inhibiting inflammatory responses by using an anti-inflammatory compound (C) as defined in claim 1. 11. A method for suppressing or inhibiting the expression of at least one cytokine in a mammal, wherein said method comprises administering to a mammal in need thereof an amount of an anti-inflammatory compound (C) as defined in claim 1 sufficient to inhibit or suppress at least one cytokine, where the specified cytokine is selected from the group consisting of IL-8, Gro-α, MIP-1, MCP-1, ENA-78 and RANTES, IL-1α, IL-1β, IL-2, IL-3, IL-6 , IL-7, IL-9, IL-12, IL-17, IL-18, IL-23, TNF-α, LT, LIF, oncostatin, IFN-α, IFN-β, IFN-γ, IL-4 , IL-10, IL-11, W-13 and TGF-β, or that at least one cytokine is selected from the group consisting of IL-8, IL-4, IL-6, IL-10, TNF-α, TNF-β, IL-17, IL-23 and IFN-γ, or that at least one of these cytokines is selected from the group consisting of IL-8, IL-1β and IFN-γ.