US20200289604A1 - Pharmaceutical compositions containing hyaluronic acid and carnosine and relative use - Google Patents

Pharmaceutical compositions containing hyaluronic acid and carnosine and relative use Download PDF

Info

Publication number
US20200289604A1
US20200289604A1 US16/753,152 US201816753152A US2020289604A1 US 20200289604 A1 US20200289604 A1 US 20200289604A1 US 201816753152 A US201816753152 A US 201816753152A US 2020289604 A1 US2020289604 A1 US 2020289604A1
Authority
US
United States
Prior art keywords
carnosine
pharmaceutical composition
composition according
kda
range
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Abandoned
Application number
US16/753,152
Other languages
English (en)
Inventor
Antonella Schiavinato
Valentina GRECO
Luciano Messina
Susanna Vaccaro
Enrico RIZZARELLI
Sebastiano SCIUTO
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Fidia Farmaceutici SpA
Original Assignee
Fidia Farmaceutici SpA
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Fidia Farmaceutici SpA filed Critical Fidia Farmaceutici SpA
Assigned to FIDIA FARMACEUTICI S.P.A. reassignment FIDIA FARMACEUTICI S.P.A. ASSIGNMENT OF ASSIGNORS INTEREST (SEE DOCUMENT FOR DETAILS). Assignors: GRECO, Valentina, MESSINA, LUCIANO, RIZZARELLI, ENRICO, SCHIAVINATO, ANTONELLA, SCIUTO, Sebastiano, VACCARO, SUSANNA
Publication of US20200289604A1 publication Critical patent/US20200289604A1/en
Abandoned legal-status Critical Current

Links

Images

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/04—Peptides having up to 20 amino acids in a fully defined sequence; Derivatives thereof
    • A61K38/05—Dipeptides
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/56—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule
    • A61K47/61—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule the organic macromolecular compound being a polysaccharide or a derivative thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/715—Polysaccharides, i.e. having more than five saccharide radicals attached to each other by glycosidic linkages; Derivatives thereof, e.g. ethers, esters
    • A61K31/726—Glycosaminoglycans, i.e. mucopolysaccharides
    • A61K31/728—Hyaluronic acid
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/54—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic compound
    • A61K47/542—Carboxylic acids, e.g. a fatty acid or an amino acid
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/62—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being a protein, peptide or polyamino acid
    • A61K47/64—Drug-peptide, drug-protein or drug-polyamino acid conjugates, i.e. the modifying agent being a peptide, protein or polyamino acid which is covalently bonded or complexed to a therapeutically active agent
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P19/00—Drugs for skeletal disorders
    • A61P19/02—Drugs for skeletal disorders for joint disorders, e.g. arthritis, arthrosis
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P19/00—Drugs for skeletal disorders
    • A61P19/08—Drugs for skeletal disorders for bone diseases, e.g. rachitism, Paget's disease
    • A61P19/10—Drugs for skeletal disorders for bone diseases, e.g. rachitism, Paget's disease for osteoporosis
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/0012—Galenical forms characterised by the site of application
    • A61K9/0014—Skin, i.e. galenical aspects of topical compositions
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/0012—Galenical forms characterised by the site of application
    • A61K9/0019—Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner
    • A61K9/0029—Parenteral nutrition; Parenteral nutrition compositions as drug carriers
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/0012—Galenical forms characterised by the site of application
    • A61K9/0053—Mouth and digestive tract, i.e. intraoral and peroral administration

Definitions

  • the present invention relates to pharmaceutical compositions containing Hyaluronic Acid and Carnosine and their relative use.
  • Osteoarthritis is a severely disabling pathology characterized by the progressive erosion of articular cartilages due to degradation of the matrix and the loss of the main cartilaginous cellular components: chondrocytes.
  • MMP Metalloprotease
  • cytokines also stimulate the synthesis of high levels of nitric oxide (responsible for death by apoptosis of chondrocytes) and, moreover, inhibit the synthesis of proteoglycans (structural components of the matrix) (Dozin B. et al., Matrix Biology, 2002, 21:449-459).
  • RA rheumatoid arthritis
  • psoriatic arthritis Arend W. P. et al., Arthritis Rheum, 1995, 38:151-160.
  • RA Rheumatoid arthritis
  • the progression of RA determines a strong inflammatory response of the synovium with a consequent swelling of its cells, an excess of synovial fluid and the development of fibrous tissue.
  • the synovium is a membrane of mesenchymal origin formed by synoviocytes (of the macrophage and fibrinoidal type) which, in the disease, undergoes hyperplasia and hypertrophy and then grows in thickness (the two/three layers of a non-pathological condition become seven or more), forming the synovial pannus that begins to peripherally erode the bone not covered by cartilage.
  • the polymorphonuclear cells with T, B lymphocytes and plasma cells move into the synovial fluid, increasing the inflammation of the affected joint, the articular cartilage affected by RA (with relative underlying bone) then undergoes thinning with progressive destruction.
  • Rheumatoid nodules are typical, that can be formed at the pulmonary level with consequent pulmonary fibrosis, pleurisy and pleuropericarditis, at the cardiac level there can be an acceleration of coronary atherosclerosis, whereas at an ocular level there can be xerophthalmia, uveitis and scleritis.
  • Amyloidosis and osteoporosis are also complications of RA (Cecil, TEXTBOOK of MEDICINE, 1988).
  • Rheumatoid arthritis affects from 0.5 to 1% of adults in the world, and commonly begins between the age of 40 and 50 years.
  • RA RA-relieving and anti-inflammatory drugs, including steroids and NSAIDs, suppress the symptoms but do not stop the progression of the condition, anti-rheumatic drugs modifying the disease (DMARD) can, however, slow it down.
  • methotrexate is generally used, an antimetabolite inhibitor of the synthesis of folic acid which, at low doses, acts as an immunosuppressant, but recently biological drugs have been introduced that act in a more selective and specific way (Canete J D et al. Expert Opinion Biol Ther, 2017, 17:1-15), such as Etanercept, a fusion protein that acts by antagonizing the pro-inflammatory TNF cytokine.
  • TNF is part of that group of cytokines responsible for the acute phase of systemic inflammation; it is involved in numerous processes such as cell proliferation, differentiation and apoptosis, carcinogenesis and viral replication. It is mostly produced by macrophages; its synthesis can be stimulated by bacterial endotoxins and be inhibited by steroids.
  • cytokine Acting on numerous organs and systems, together with other cytokines, it promotes the inflammatory response which in turn ignites many pathologies (including autoimmune diseases) such as RA and OA, Crohn's disease, psoriasis and asthma; this cytokine is also capable of activating osteoclasts and therefore inducing bone resorption, it is capable of stimulating the macrophage production of molecules with an oxidizing action, it is involved in particular pathologies of the cardiovascular system participating in the formation of venous thrombi, in the pathogenesis of atherosclerosis and vasculitis (Alam J., Biomed Pharmacother, 2017, 92:615-633), and finally it is capable of increasing the resistance of tissues to insulin favouring the onset of type II diabetes (Nicolau J et al., Joint Bone Spine, 2017, 84 (4):411-416).
  • OA and RA there are various types of drugs which effective in slowing down the progression of the above-mentioned pathologies or in the treatment of related symptoms, however they can have important side-effects, often toxic, both as synthetic molecules (therefore not natural), for the pharmacological treatment regimen that must be sustained for very long periods of time, and also for the method of administration, generally parenteral, but above all oral and therefore involving the whole organism even if, in many cases, only the inflamed joint is to be treated.
  • the objective of the present invention is therefore to identify pharmaceutical compositions comprising hyaluronic acid (HA)/carnosine conjugate particularly effective for use in the treatment and prevention of OA and in the treatment of RA (with all the consequent diseases caused directly by RA or indirectly related to/depending on RA), preferably by oral administration (above all when the disease has acquired clear systemic manifestations) and/or by intra-articular administration in the joint in an early stage of the disease, or in an intensification localized phase in the same joint.
  • HA hyaluronic acid
  • carnosine conjugate particularly effective for use in the treatment and prevention of OA and in the treatment of RA (with all the consequent diseases caused directly by RA or indirectly related to/depending on RA)
  • oral administration above all when the disease has acquired clear systemic manifestations
  • intra-articular administration in the joint in an early stage of the disease or in an intensification localized phase in the same joint.
  • Carnosine is a dipeptide obtained from the condensation reaction between ⁇ -alanine and L-histidine, in the body. It is found in large quantities in the muscles and in the brain; scientific literature is unanimous in giving it an anti-oxidant, anti-radical and anti-inflammatory activity (Budzen S, et al., Adv Clin Exp Med, 2013, 22 (5): 739-44).
  • the cyclodextrin stabilizes the dipeptide, protecting it from the hydrolytic activity of carnosinase (Schaschke N. et al., JACS, 1998, 120 (28):7030-7038), thus allowing the above-mentioned molecule to exert its biological activity.
  • the carnosine conjugate with trehalose was subsequently patented as a system with an anti-oxidant, anti-glycating and anti-aggregating activity (EP1860116).
  • the derivative of HA is also known, which is obtained by conjugating carnosine with said polysaccharide by an amide bond involving the carboxyl of HA and the amino group of the dipeptide (WO2016016847).
  • this derivative could only be derivatized up to 25% (mole/mole ratio between HA and carnosine).
  • the present invention describes a composition comprising the amide derivative of HA with carnosine with a degree of derivatization higher than 25%, therefore a conjugate whose synthesis was not considered possible by skilled persons in the field based on the teaching of the state of the art.
  • HA is a hetero-polysaccharide composed of alternating residues of D-glucuronic acid and N-acetyl-D-glucosamine. It is a linear chain polymer with a molecular weight ranging from 50,000 to 13 ⁇ 106 Da, depending on the source from which it is obtained and the preparation methods used. It is present in nature in pericellular gels, in the fundamental substance of the connective tissue of vertebrate organisms, in the synovial fluid of the joints, in the vitreous humor and in the umbilical cord.
  • HA therefore plays an important role in the biological organism, both as a mechanical support of the cells of many tissues such as the skin, tendons, muscles and cartilage, and also as an active agent capable of modulating (through the CD44 receptor) many processes relating to cell physiology and biology, such as, for example, proliferation, migration, cell differentiation and angiogenesis; its role in maintaining the hydration of the tissues and lubrication of the joints is also known.
  • HA is in fact a polysaccharide that has particular viscoelastic properties, synthesized and secreted in the joint cavity mainly by synoviocytes (Asari A. et al., Arch. Histol.
  • HA acts as a viscous lubricant, whereas during fast movements, it absorbs with elastic properties any possible traumas or microtraumas that may strike the joint.
  • Balazs first suggested the possibility of modifying the evolution of the osteoarthritic process through the contribution of exogenous HA (especially with a high MW) directly in the joint cavity, and thanks to this treatment therapy it was possible to demonstrate the protective effect of HA against the degeneration involved in the cartilage of a joint damaged by inflammatory diseases or trauma.
  • the object of the present invention relates to a pharmaceutical composition
  • a pharmaceutical composition comprising a hyaluronic acid (HA)/carnosine conjugate with a direct amide bond between the carboxyl group of HA and the amino group of carnosine for use in the prevention and treatment of OA in the treatment of RA and pathologies caused directly by RA or indirectly related to/depending on RA, in particular for use in the treatment of diseases caused directly by RA selected from the group comprising pulmonary fibrosis, pleurisy and pleuropycarditis, xerophthalmia, uveitis and scleritis, or indirectly related to/depending on RA selected from the group including premature pathological aging, heart attack, stroke, arterosclerosis and coronary atherosclerosis, arterial hypertension, dementia, diabetes, osteoporosis and amyloidosis, cancer, proteinuria and nephritis, gastric lesions, cataracts, psoriatic arthritis, gastritis
  • compositions for use according to the present invention preferably comprise the HA/carnosine conjugate with a degree of derivatization (amidation) of the carboxyl group of HA (DS) higher than 25% (from 25% to 100%), preferably ranging from 30% to 100%, more preferably ranging from 30% to 60%, and even more preferably varying within the range of 35 ⁇ 3% (i.e. from 32% to 38%) to 45 ⁇ 5% (i.e. from 40% to 50%).
  • Said amide derivative of HA with carnosine is obtained by covalently combining the dipeptide (i.e. carnosine) with hyaluronic acid (HA) by forming a direct amide bond (without the aid of permanent spacers) between carboxyl of HA and amine group of carnosine.
  • the Applicant has in fact surprisingly discovered how the conjugate object of the invention, has unexpected synergistic effects between carnosine and HA in the curative treatment of RA and OA, achieving a significant clinical/histological improvement of the above-mentioned pathologies, with a significant reduction in plasma indicators of both inflammation and lipid oxidation.
  • the degree of derivatization of the conjugate object of the present invention is measured by the method described in the following Example 4.
  • the HA used for the synthesis of the HA/carnosine conjugate of the present invention can derive from any source, for example, by extraction from rooster combs (EP138572), by fermentation (from Streptococcus Equi or Zooepidemicus, EP716688), or by biosynthesis (from Bacillus, EP2614087), and can have an average molecular weight (MW) ranging from 400 to 3 ⁇ 106 Da, in particular from 1 ⁇ 10 5 Da to 1 ⁇ 10 6 Da, even more particularly within the range of 130-220 kDa and/or within the range of 500-750 kDa.
  • MW average molecular weight
  • compositions wherein the degree of derivatization of HA varies within the range of 30 to 60% and the average molecular weight of HA ranges from 1 ⁇ 10 5 Da to 1 ⁇ 10 6 Da; pharmaceutical compositions wherein the DS of HA varies within the range of 30 to 60% and the average molecular weight of HA is within the range of 130-220 kDa, or 500-750 kDa and relative mixtures, are more preferred.
  • compositions wherein the DS of HA varies within the range of 31 to 34% (35 ⁇ 3%) or within the range of 40 to 50% (45 ⁇ 5%) and relative mixtures, and the average molecular weight of HA is within the range of 130-220 kDa or 500-750 kDa and relative mixtures; in particular, pharmaceutical compositions are preferred wherein, when the DS of HA varies within the range of 35 ⁇ 3%, the average molecular weight of HA is within the range of 500-750 kDa, and when the DS of HA varies within the range of 45 ⁇ 5%, the average molecular weight of HA is within the range of 130-220 kDa.
  • average molecular weight refers to the weight average MW calculated with the “intrinsic viscosity” method (Terbojevich et al., Carbohydr Res, 1986, 363-377).
  • the hyaluronic acid used for the synthesis of the conjugate in question is selected from the sodium salt of HA (EP138572), preferable for the synthesis of the HA/carnosine conjugate up to a DS of about 40% (therefore from 25% to 40%), and the tetrabutylammonium (TBA) salt of HA (EP216453), preferable for the synthesis of the HA/carnosine conjugate with a DS higher than 40% (therefore from 40% to 100%), the final product (described in the preparation examples provided hereunder) being (preferably) prepared as the sodium salt of the HA/carnosine conjugate, regardless of the HA salt used as reagent.
  • TSA tetrabutylammonium
  • a further object of the invention relates to biomaterials consisting of conjugated HA derivatives (again through the amide bond) with carnosine, in the modes and in the degrees described for HA, defined herein as “HA/carnosine biomaterials”.
  • HA derivatives that can be used in the formation of the above-mentioned biomaterials are listed hereunder:
  • HYAFF® esters of HA with alcohols of the aliphatic, araliphatic, cycloaliphatic, aromatic, cyclic and heterocyclic series, with an esterification percentage that can vary depending on the type and length of the alcohol used, preferably from 1 to 70%, whereas the remaining percentage of non-esterified HA can be salified with organic and/or inorganic bases (EP216453);
  • HYADD® amides of HA with amines of the aliphatic, araliphatic, cycloaliphatic, aromatic, cyclic and heterocyclic series, with an amidation percentage ranging from 0.1 to 50%, whereas the remaining percentage of HA not subjected to amidation can he salified with organic and/or inorganic bases (EP1095064);
  • ACP® internal esters of HA with an esterification percentage not higher than 20%, preferably ranging from 0.05 to 10% of esterification, whereas the remaining percentage of non-esterified HA can be salified with organic and/or inorganic bases (EP341745);
  • HYOXX® derivatives of percarboxylates of HA obtained by the oxidation of the primary hydroxyl of the N-acetyl-glucosamine fraction with a percarboxylation degree ranging from 0.1 to 100% and, preferably, from 25 to 75%.
  • the carboxyl groups of HA can be salified with organic and/or inorganic bases (EP1339753).
  • HA/carnosine conjugate it is also possible to start from the HA/carnosine conjugate to synthesize (according to what is known to skilled persons in the field) the above-mentioned HA/carnosine biomaterials further derivatizing the HA: it is therefore possible to use the conjugate in question as starting material (instead of the HA sodium salt or TBA salt) to proceed with the synthesis of HYAFF, ACP, HYADD, HYOXX or sulfated derivatives according to what is known to skilled persons in the field, to obtain products previously defined as HA/carnosine biomaterials.
  • Esters and amides are, among the derivatives of HA, those that are particularly important in the formation process of the biomaterials object of the invention; particularly interesting under this profile are benzyl esters (HYAFF) with an esterification percentage preferably ranging from 5 to 50% and the hexadecyl amide of HA (HYADD4) with a derivatization degree of up to 5% (measured in HPLC), this amide has proved to be particularly relevant for its role as visco-elastic gel that can be clinically used as a synovial substitute in the case of joint injury (EP1853279).
  • HYAFF benzyl esters
  • HYADD4 hexadecyl amide of HA
  • HYAFF is a completely biocompatible biodegradable polymer
  • EP618817 ester derivatives of HA
  • EP618817 ester derivatives of HA
  • non-woven form or as a sponge can constitute a three-dimensional matrix that can be used in the dermatological and orthopedic fields.
  • ACP derivatives have been used successfully as skin fillers and as anti-adhesion gels (EP0850074), whereas for the sulfated derivative of HA, its anti-inflammatory effects are being studied as a topical component of compositions for dermatological use (EP2429515), or its protective/regenerative effect of the cartilage matrix in the case of oral or intra-articular compositions (EP2021078, EP2786782).
  • the new biomaterials formed by the amide bond of the carboxyl of the above-mentioned HA derivatives with carnosine find, according to the present invention, effective use in the treatment of RA (in addition to all diseases caused directly by RA or indirectly related to/depending on RA) and OA (and also in the prevention of OA), as biomaterials processed in the form of fibers, gels, hydrogels, microspheres, sponges, woven or non-woven fabrics, films.
  • RA rheumatoid nodules
  • pulmonary fibrosis pulmonary fibrosis
  • pleurisy pulmonary fibrosis
  • pleuropericarditis pulmonary fibrosis
  • xerophthalmia uveitis and scleritis.
  • pathologies indirectly related to/depending on RA are defined herein as pathologies that have been indirectly caused by RA as they are linked to serious situations of altered immune profile and/or to conditions of high oxidative stress.
  • Oxidative stress can therefore be considered as a pathological condition caused by the breaking of a delicate balance between the production and disposal of free radicals.
  • Oxidative stress is currently considered as actively contributing to the onset of RA and, consequently, other important diseases that frequently arise contextually and/or depend on RA, such as premature pathological aging, heart attack, stroke, arteriosclerosis and coronary atherosclerosis, arterial hypertension, dementia, diabetes, osteoporosis and amyloidosis, many forms of cancer, certain hepatic or renal diseases such as proteinuria and nephritis, gastric lesions, cataracts.
  • Diseases linked to an altered immune profile include psoriatic arthritis, gastritis, vasculitis, type I diabetes and autoimmune diseases including, for example, autoimmune thyroiditis.
  • compositions comprising HA/carnosine conjugates and HA/carnosine biomaterials are described for a topical, oral, intra-articular, parenteral or surgical application, in all fields in which said administration is required in relation to the prevention or treatment of the pathologies described above and related/dependent diseases, and can be associated with pharmacologically and/or biologically active substances such as, for example, steroids, anti-inflammatory cytokines, interferon, growth factors (such as, for example, BMP2 and BMP7), NSAIDs, and/or with controlled drug delivery systems such as, for example, cyclodextrins, and/or natural polymers, such as HA and its derivatives, preferably with the amide derivative of HA with hexadecylamine called HYADD4 (EP1095064), or of a synthetic nature.
  • pharmacologically and/or biologically active substances such as, for example, steroids, anti-inflammatory cytokines, interferon, growth factors (such as, for example, BMP
  • HA/carnosine conjugates object of the invention whose conjugation process between carnosine methyl ester and hyaluronic acid sodium salt or tetrabutylammonium salt takes place in a medium comprising DMSO or DMSO/H 2 O as the main solvent.
  • the synthesis therefore takes place in a dipolar solvent, aprotic and strongly hygroscopic, at room temperature.
  • the synthesis in THF according to the state of the art was effected in an aprotic, volatile solvent, having a lower polarity, and was therefore carried out at 4° C.
  • the reaction was carried out in an anhydrous environment.
  • 1.5 g of carnosine were treated, under stirring, inside a 250 ml flask, with 50 ml of an acetyl chloride solution in anhydrous methanol (pre-mixed) in a 1:20 ratio (v/v) for 12 hours, after which about 90% of the solvent was removed by evaporation under vacuum.
  • 20 ml of anhydrous methanol were added to the reaction residue and again about 90% of the solvent was removed by evaporation. The operation was repeated until all the (which was formed during the reaction) had been removed; the product was then brought to dryness under vacuum.
  • the product was subjected to control by electrophoresis on paper and the various components of the sample were detected using ninhydrin.
  • hyaluronic acid sodium salt (HANa) with a MW of 700 kDa were introduced into a reactor in which 80 ml of a mixture of H 2 O:DMSO (in a ratio of 1:1 v/v) were subsequently added under constant stirring (at room temperature) for at least 4 hours.
  • HTBA hyaluronic acid tetrabutylammonium salt
  • TAEA Tris [2-(2-methoxyethoxy) ethyl]amine
  • HOOBt 3-hydroxy-1,2,3-benzotriazin-4(3H)-one
  • DMSO DMSO containing 240 mg of N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride
  • Example 2 showed a degree of derivatization to the carboxyl of HA of 35% mol/mol, whereas the sample of Example 3 showed a degree of derivatization equal to 50% mol/mol.
  • CIA has for many years been recognized as a valid experimental model of RA in mice or rats, as it determines humoral, cellular, histological and pathological characteristics of RA in the animal (Holmdahl et al., Immunological reviews, 1990, 118: 193-232). Following the induction of CIA in mice, in fact, a large number of activated neutrophils/macrophages/lymphocytes are present in its joints, with the consequent production of the synovial pannus of RA.
  • Type II chicken collagen (CII) was dissolved in acetic acid 0.01M at a concentration of 2 mg/ml, the complete Freund's adjuvant (CFA) was then prepared by the addition of Mycobacterium tuberculosis H37Ra (2 mg/ml) to the incomplete adjuvant (composed of water/mineral oil emulsion), the two components (CII and CFA) were then mixed and emulsified in the same ratio v/v.
  • CFA complete Freund's adjuvant
  • mice Nine-week old DBA/IJ male mice were used, about 30 g each, 20 mice per group.
  • Group 1 CIA-control: the animals were treated by CIA induction and subsequently with distilled water administered orally every 24 hours starting from the 25th day after the CIA until the 35th day, end of the experimentation.
  • Group 2 CIA+HA/carnosine conjugate: the animals were treated by CIA induction and subsequently with the above-mentioned conjugate every 24 hours starting from the 25th day after the CIA until the 35th day.
  • the conjugate used for the test had been prepared according to Example 3, it was therefore derivatized at 50% mol/mol, the dose, administered orally, was 81 mg/kg of conjugate (20 mg of carnosine and 61 mg of HA).
  • Group 3 CIA+HA/carnosine mixture: the animals were treated by CIA induction and subsequently with HA/carnosine every 24 hours starting from the 25th day after the CIA until the 35th day.
  • This preparation consisted of a mixture of HA sodium salt mixed with carnosine in water, administered orally at a dose of 20 mg/kg of carnosine with 61 mg/kg of HA.
  • the total CIA development index (defined as Arthritic Paw score) was calculated by summing the values obtained (and evaluated as above) for each paw.
  • the clinical severity index is given by the volume of the paws (measured every two days by a plethysmometer) which progressively increased throughout the whole analysis period based on the severity of the disease: foot increase.
  • the animals were sacrificed, the treated paws removed (including the knee joints) and fixed in 10% formalin; subsequently decalcified and soaked in paraffin for the preparation of 5 ⁇ m microtome sections, then stained with hematoxylin/eosin.
  • the optical microscope analysis was effected following the following Histological score:
  • TNF- ⁇ levels were measured in animal plasma at the end of the experimentation using the ELISA kit of Calbiochem-Novabiochem Corporation, IT, for determining the TNF- ⁇ levels (reading limit 10 pg/ml).
  • the attack by free radicals of the lipids present in the biological membranes determines the start of an oxygen-dependent deterioration process (lipid peroxidation) which leads to the impairment of the integrity of the biological membranes, and to the formation of oxidized lipid proteins and lipid peroxides, with by-products such as aldehydes including malondi aldehyde (MDA). All of these processes lead to oxidative stress.
  • lipid peroxidation oxygen-dependent deterioration process
  • MDA malondi aldehyde
  • FIG. 2 shows the trend of the foot increase: also in this case the treatment with the conjugate object of the invention confirms the surprising effect of FIG. 1 , in which, at the end of the experiment, the swelling of the paw treated with the conjugate was significantly lower versus both the untreated control and also versus the group being treated with the HA/Carnosine mixture.
  • FIG. 3 at the 35th day (end) of the experimentation, the histological evaluation of CIA-control animals revealed clear and important histological signs of the development of a severe form of RA, with hone erosion and moderate/severe necrosis of the joint tissues. Treatment with the HA/Carnosine conjugate, on the other hand, significantly reduced this erosion both versus the control and versus the group treated with the HA/Carnosine mixture.
  • the plasma level of the pro-inflammatory cytokine TNF- ⁇ was analyzed in all the animals subjected to CIA and not (Sham sample, i.e. in which CIA had not been induced to have the plasma TNF value of non-operated healthy animals as a comparison).
  • the value of the above-mentioned inflammatory cytokine in the CIA-control group was extremely high, almost 5 times higher than the Sham control, whereas the treatment with the conjugate object of the invention more than halved the TNF- ⁇ value, also significantly reducing it versus the group to which the HA/Carnosine mixture had been administered.
  • MIA Monosodium Iodoacetate
  • OA was induced in the posterior knee joints of the rats being tested, by the intra-articular injection of MIA: 25 ⁇ l of saline containing 1 mg/kg of MIA were injected into the joint (knee) of the right paw of each animal (under treatment) through the infrapatal ligament, whereas the left joint received an equal volume of saline 0.9% (Sagar D R et al, Ann. Rheum. Dis., 2013, 73: 1558-1565).
  • Group 1 MIA-control: the animals were treated by MIA induction and subsequently with 25 ⁇ l of distilled water administered intra-articularly in the right knee at day 7, 10, 13, 16 and 19 after MIA, on the 21st day, the experimentation ended with euthanasia of the animals.
  • Group 2 MIA+HA/carnosine conjugate: the animals were treated by MIA induction and subsequently with 25 ⁇ l of the conjugate prepared according to Example 2, then derivatized at 35% mol/mol, intra-articularly in the right knee at day 7, 10, 13, and 16 19 after MIA until the 21st day.
  • the 25 ⁇ l of HA/carnosine conjugate contained 250 ⁇ g of conjugate, i.e. 45.75 ⁇ g of carnosine and 204.25 ⁇ g of HA.
  • Group 3 MIA+HA/carnosine mixture: the animals were treated by MIA induction and subsequently with 25 ⁇ l of HA and sine intra-articularly in the right knee at day 7, 10, 13, 16 and 19 after MIA until the 21st day.
  • This preparation consisted of a mixture of HA sodium salt mixed with carnosine in water, therefore 25 ⁇ l of this preparation contained 45.75 ⁇ g of carnosine and 204.25 ⁇ g of HA.
  • the TNF- ⁇ levels were measured in the plasma of the animals using the ELISA kit of Calbiochem-Novabiochem Corporation, IT, for determining the TNF- ⁇ levels (reading limit 10 pg/ml).
  • FIG. 6 shows the results obtained in the treatment groups and in the Sham group, defined as a group in which the animals had not been subjected to treatment with MIA: also in this case, the plasma level of the TNF cytokine of the MIA-control group proved to be extremely high, whereas the treatment with the HA/Carnosine conjugate lowered the TNF- ⁇ value significantly not only versus this control, but also versus the group to which the HA/Carnosine mixture had been administered that did not show any efficacy, demonstrating once again the striking effect of the conjugate object of the invention, and therefore its level of derivatization which is particularly high with respect to the state of the art.
  • mice Male Sprague-Dawley rats, of about 230 g each, were used, 20 rats per group.
  • Group 1 MIA-control: the animals were treated by MIA induction and subsequently with distilled water administered orally every 24 hours from the 3rd day after MIA until the 20th day, end of the experimentation.
  • Group 2 MIA+HA/carnosine conjugate: the animals were treated by MIA induction and subsequently with the above-mentioned conjugate every 24 hours from the 3rd day after MIA until the 20th day.
  • the conjugate used for the experimentation was prepared according to Example 2, it was then derivatized at 35% mol/mol, the dose, administered orally, was equal to 88.5 mg/kg of conjugate (16.2 mg of carnosine and 72.3 mg of HA).
  • Group 3 MIA+HA/carnosine mixture: the animals were treated by MIA induction and subsequently with HA/carnosine every 24 hours starting from the 3rd day after MIA up to the 20th day.
  • This preparation consisted of a mixture of HA sodium salt mixed with carnosine in water, administered orally at a dose of 16.2 mg/kg of carnosine with 72.3 mg/kg of HA.
  • Group 4 MIA+naproxen: the animals were treated by MIA induction and subsequently with naproxen administered orally at a dose of 10 mg/kg every 24 hours starting from the 3rd day after MIA until the 20th day.
  • the animals were sacrificed, the treated legs removed (including the knee joints) and fixed in 10% formalin. They were subsequently decalcified and soaked in paraffin for the preparation of 5 ⁇ m microtome sections, then stained with hematoxylin/eosin.
  • the optical microscope analysis was performed following a modified Histological score by Mankin, with a score range of 0-12, i.e. from a normal cellular organization to complete disorganization and hypocellularity; (Mankin H J et al., J Bone Joint Surg Am., 1971, 53 (3): 523-537).
  • the histological examination of the samples of group 1 showed irregularities in the organization of the tissues with fibrillation of the surface layer, a reduced cellularity and degeneration of the cartilage layer.
  • Oral treatment with the HA/carnosine conjugate very significantly reduced histological damage and cartilage degeneration, totally comparable to the effect elicited by naproxen (known NSAIDs of documented effectiveness), whereas treatment with the HA/carnosine mixture did not lead to any histological improvement with respect to the untreated samples of Group 1 (MIA-control) ( FIG. 7 ).

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • General Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Animal Behavior & Ethology (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Engineering & Computer Science (AREA)
  • Epidemiology (AREA)
  • Rheumatology (AREA)
  • General Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Physical Education & Sports Medicine (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Orthopedic Medicine & Surgery (AREA)
  • Immunology (AREA)
  • Pain & Pain Management (AREA)
  • Molecular Biology (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Dermatology (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
US16/753,152 2017-10-03 2018-10-03 Pharmaceutical compositions containing hyaluronic acid and carnosine and relative use Abandoned US20200289604A1 (en)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
IT102017000110784A IT201700110784A1 (it) 2017-10-03 2017-10-03 Composizioni farmaceutiche contenenti Acido Ialuronico e Carnosina e relativo uso
IT102017000110784 2017-10-03
PCT/IB2018/057697 WO2019069258A1 (en) 2017-10-03 2018-10-03 PHARMACEUTICAL COMPOSITIONS CONTAINING HYALURONIC ACID AND CARNOSINE AND USE THEREOF

Related Parent Applications (1)

Application Number Title Priority Date Filing Date
PCT/IB2018/057697 A-371-Of-International WO2019069258A1 (en) 2017-10-03 2018-10-03 PHARMACEUTICAL COMPOSITIONS CONTAINING HYALURONIC ACID AND CARNOSINE AND USE THEREOF

Related Child Applications (1)

Application Number Title Priority Date Filing Date
US17/482,696 Continuation US20220031797A1 (en) 2017-10-03 2021-09-23 Pharmaceutical compositions containing hyaluronic acid and carnosine and relative use

Publications (1)

Publication Number Publication Date
US20200289604A1 true US20200289604A1 (en) 2020-09-17

Family

ID=61024973

Family Applications (2)

Application Number Title Priority Date Filing Date
US16/753,152 Abandoned US20200289604A1 (en) 2017-10-03 2018-10-03 Pharmaceutical compositions containing hyaluronic acid and carnosine and relative use
US17/482,696 Pending US20220031797A1 (en) 2017-10-03 2021-09-23 Pharmaceutical compositions containing hyaluronic acid and carnosine and relative use

Family Applications After (1)

Application Number Title Priority Date Filing Date
US17/482,696 Pending US20220031797A1 (en) 2017-10-03 2021-09-23 Pharmaceutical compositions containing hyaluronic acid and carnosine and relative use

Country Status (18)

Country Link
US (2) US20200289604A1 (fi)
EP (1) EP3691691B1 (fi)
KR (1) KR20200091380A (fi)
CN (1) CN111163808A (fi)
AR (1) AR114337A1 (fi)
CA (1) CA3073797A1 (fi)
DK (1) DK3691691T3 (fi)
EA (1) EA202090524A1 (fi)
FI (1) FI3691691T3 (fi)
HR (1) HRP20240187T1 (fi)
IT (1) IT201700110784A1 (fi)
LT (1) LT3691691T (fi)
PL (1) PL3691691T3 (fi)
PT (1) PT3691691T (fi)
RS (1) RS65191B1 (fi)
SI (1) SI3691691T1 (fi)
WO (1) WO2019069258A1 (fi)
ZA (1) ZA202001174B (fi)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN117700583A (zh) * 2023-12-22 2024-03-15 珠海瑞玞生物工程有限公司 一种透明质酸基生物材料的制备方法

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
IT202000014017A1 (it) 2020-06-11 2021-12-11 Fidia Farm Spa Nuovi derivati ottenuti da acido ialuronico e carnosina

Family Cites Families (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
MXPA05010778A (es) * 2003-04-09 2005-12-12 Genentech Inc Terapia para enfermedad autoinmune en un paciente con una respuesta inadecuada a un inhibidor tnf-alfa.
ITPD20040053A1 (it) * 2004-02-27 2004-05-27 Fidia Advanced Biopolymers Srl Biomateriali costituiti da derivati dell'acido ialuronico come nuova terapia di cura per la protezione e la la riparazione della cartilagine articolare danneggiata per osteoartrosi
KR101234476B1 (ko) * 2004-03-05 2013-02-18 덴끼 가가꾸 고교 가부시키가이샤 히알루론산/메토트렉세이트 화합물
ITPD20050056A1 (it) * 2005-03-02 2006-09-03 Fidia Farmaceutici Derivati ammidici del'acido ialuronico in osteoartrosi
IT1391251B1 (it) * 2008-10-15 2011-12-01 Innovative Nutrition & Pharma Srl Composizione per il trattamento delle patologie articolari
US20120321603A1 (en) * 2011-06-16 2012-12-20 Nayan Patel Composition and method for stabilization and delivery of therapeutic molecules
PL3174555T3 (pl) * 2014-07-31 2019-08-30 Sebastiano SCIUTO Pochodne otrzymane z kwasu hialuronowego i karnozyny

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN117700583A (zh) * 2023-12-22 2024-03-15 珠海瑞玞生物工程有限公司 一种透明质酸基生物材料的制备方法

Also Published As

Publication number Publication date
HRP20240187T1 (hr) 2024-05-10
US20220031797A1 (en) 2022-02-03
LT3691691T (lt) 2024-02-26
RS65191B1 (sr) 2024-03-29
PL3691691T3 (pl) 2024-04-29
ZA202001174B (en) 2021-08-25
EP3691691A1 (en) 2020-08-12
EA202090524A1 (ru) 2020-05-28
AR114337A1 (es) 2020-08-26
EP3691691B1 (en) 2023-12-13
KR20200091380A (ko) 2020-07-30
IT201700110784A1 (it) 2019-04-03
WO2019069258A1 (en) 2019-04-11
CA3073797A1 (en) 2019-04-11
CN111163808A (zh) 2020-05-15
DK3691691T3 (da) 2024-02-05
PT3691691T (pt) 2024-01-31
FI3691691T3 (fi) 2024-02-08
SI3691691T1 (sl) 2024-03-29

Similar Documents

Publication Publication Date Title
US20220031797A1 (en) Pharmaceutical compositions containing hyaluronic acid and carnosine and relative use
US9301972B2 (en) Hyaluronic acid derivatives
US20050164980A1 (en) Hyaluronic acid modification product
US8202986B2 (en) Branched hyaluronic acid and method of manufacture
AU2002221528B2 (en) Use of hyaluronic acid derivatives for the prevention of inflammatory arthritis
CA2598331A1 (en) Amide derivatives of hyaluronic acid in osteoarthrosis
BRPI0708128A2 (pt) derivado de ácido hialurÈnico, processo para produção de um derivado de ácido hialurÈnico, composição, artigo cosmético, artigo sanitário, médico ou cirúrgico, cápsula ou microcápsula de medicamento, métodos para realizar procedimentos em oftalmologia, no tratamento de osteoartrite, e no tratamento de cáncer, para realizar administração transdérmica e dérmica de um agente farmacologicamente ativo, e administração dérmica de um cosmético, e para realizar procedimentos no tratamento de perda de cabelo ou calvìcie, métodos oftalmológico, de tratamento de osteoartrite, de uma ferida, e de perda de cabelo ou calvìcie, e, uso de um derivado de ácido hialurÈnico
JP4990446B2 (ja) 関節症治療用注入剤
ES2281265B1 (es) Composiciones para el tratamiento de la artrosis.
AU2010247814B2 (en) Lipid-polymer conjugates, their preparation and uses thereof
Kaderli et al. A novel oxido-viscosifying Hyaluronic Acid-antioxidant conjugate for osteoarthritis therapy: Biocompatibility assessments
EA042176B1 (ru) Фармацевтические композиции, содержащие гиалуроновую кислоту и карнозин, и их соответствующее применение
US20170216343A1 (en) Viscosupplement composition comprising ulvan for treating arthritis
Twitchell Restoring Joint Lubrication: The Development of a Biomimetic Lubricin Peptidoglycan to Reduce Friction at the Articular Carilage Surface
Cai Glycosaminoglycans and their binding proteins: Biochemical studies and development of potential wound healing biomaterials
Patel et al. Hyaluronic Acid (Hyaluronan): A Review on Pharmacokinetics and its Application
WO2014164723A1 (en) Short-chain fatty acid hexosamine analogs and their use in tissue engineering applications

Legal Events

Date Code Title Description
AS Assignment

Owner name: FIDIA FARMACEUTICI S.P.A., ITALY

Free format text: ASSIGNMENT OF ASSIGNORS INTEREST;ASSIGNORS:SCHIAVINATO, ANTONELLA;GRECO, VALENTINA;MESSINA, LUCIANO;AND OTHERS;REEL/FRAME:052302/0527

Effective date: 20200224

STPP Information on status: patent application and granting procedure in general

Free format text: APPLICATION DISPATCHED FROM PREEXAM, NOT YET DOCKETED

STPP Information on status: patent application and granting procedure in general

Free format text: DOCKETED NEW CASE - READY FOR EXAMINATION

STPP Information on status: patent application and granting procedure in general

Free format text: NON FINAL ACTION MAILED

STPP Information on status: patent application and granting procedure in general

Free format text: RESPONSE TO NON-FINAL OFFICE ACTION ENTERED AND FORWARDED TO EXAMINER

STPP Information on status: patent application and granting procedure in general

Free format text: NON FINAL ACTION MAILED

STCB Information on status: application discontinuation

Free format text: ABANDONED -- FAILURE TO RESPOND TO AN OFFICE ACTION