Inflammation-modulating, antifibrotic, fibrolytic and tissue-regeneration inducing composition based on a mixture of matricins (collagenocins and elastocins) and polymeric type i atelocollagen and method for producing same

Abstract

The present invention relates to a process for preparing a chemical composition based on a mixture of matricins (collagenocins, elastocins) and polymeric type I atelocollagen. The invention further relates to the composition and use thereof for modulating inflammation, inducing peripheral immune tolerance, imparting antifibrotic and fibrolytic effects, and inducing tissue regeneration, with potential applicability in conditions such as osteoarthritis (OA), rotator cuff tendinosis, epicondylitis, bursitis, tenosynovitis, tendinitis, degenerative tendinosis, partial tendon rupture, rheumatoid arthritis (RA), scleroderma (SSc), scarring, keloid or hypertrophic scarring, pulmonary fibrosis, chronic obstructive pulmonary emphysema, liver cirrhosis, valve diseases, ulcers in diabetic patients, pressure ulcers, tendon sclerosis, fracture healing, achalasia and other similar conditions, the common denominator being the presence of acute and chronic inflammation, hyperinflammation, cytokine release syndrome and fibrosis.

Description

[0001] A COMPOSITION THAT MODULATES INFLAMMATORY, ANTIFIBROTICS, FIBROLYTIC AND INDUCES TISSUE REGENERATION BASED ON A MIXTURE OF MATRICINS (COLLAGENOCINS AND ELASTOCINS) AND POLYMERIC TYPE I ATELOPEPTIDE COLLAGEN AND A METHOD FOR OBTAINING THE SAME

[0002] Technical Field

[0003] The present invention relates to the pharmaceutical field since it provides a composition based on a mixture of:

[0004] a) matricins [collagenocins (matricins obtained by heat treatment of native type I collagen of porcine origin) and elastocins (matricins obtained by heat treatment of bovine type I elastin)], and b) atelopeptide type I polymeric collagen with low molecular weight povidone.

[0005] More particularly, the present invention relates to the identification of the matrix peptides that participate in the composition that produces the technical effects of the invention, which include the negative regulation of inflammation.

[0006] Background of the Invention

[0007] The most important technical background to the invention is that of the inventor herself in the following documents and of the applicant herself, namely, patent No. MX 214259 B which refers to a composition based on collagen-polyvinylpyrrolidone or polymerized collagen with an anti-fibrotic, regenerative effect, applicable in various conditions that have as a common denominator, the presence of fibrosis and reparative processes.

[0008] Patent No. MX 264089 B refers to the use of a collagen-polyvinylpyrrolidone complex to prepare a joint inflammation modulator drug for the treatment of osteoarthritis (OA).

[0009] Patent No. MX 222625 B refers to a composition comprising a mixture of collagen hydrolysate and elastin hydrolysate.

[0010] The present invention was developed from the discovery that:

[0011] • higher concentration of collagen-polyvinylpyrrolidone, greater regulation of inflammation, and

[0012] • A higher concentration of hydrolyzed peptides, i.e., collagen and elastin, leads to greater regulation of tissue repair.

[0013] The present invention enhances the effect of collagen-polyvinylpyrrolidone with peptides resulting from the hydrolysis of elastin and collagen. Thus, the invention relates to a composition based on a mixture of matricins [collagenocins (matricins obtained by heat treatment of native type I collagen of porcine origin) and elastocins (matricins obtained by heat treatment of bovine type I elastin)] and polymeric atelopeptide type I collagen with low molecular weight povidone.The descriptions that are exemplified below show a superior effect of the mixture in inducing the synthesis of proteoglycans and type II collagen, in the proliferation of chondrocytes, in preserving the integrity of the joint cavity and cartilage, in increasing the number of CD4+ regulatory T cells, in decreasing some mediators of inflammation (IL-8, TNF-α), Thl7 cells, type I collagen and MMP-13, compared to the previous art.

[0014] Until three decades ago, extracellular matrix (ECM) macromolecules were considered solely as an essential part of tissue structure and integrity—that is, as the structural and cementing material that formed the scaffold for cell adhesion and migration. However, it is now clear that ECM components regulate important cellular functions, including morphology, cell phenotype, differentiation, migration, mitogenic activity, cell activation, apoptosis, and the synthesis of matrix macromolecules and cytokines. Nevertheless, a new biological concept has emerged in the last decade: the bioactivity of cytokines, proteins, and peptides (matricins) of the ECM are interdependent in their effects and their respective synthesis.For example, cytokines affect the expression of the extracellular matrix (ECM), cellular receptors for matrix components, and the enzymes that induce its turnover. In turn, ECM proteins (ECMs), in their "native" or modified form, modulate the synthesis of cytokines and their respective cell surface receptors. Similarly, the effects of cytokines can be mediated by ECM proteins and peptides (matricins), whose synthesis is regulated by the cytokines themselves. Thus, cytokines bind to ECMs to be presented to the receptor in an optimal state, and the ECM modulates the cellular response to cytokine stimulation.

[0015] Based on the above, peptides and PMECs in their "native" and modified forms have gained importance over the last two decades as potential biotherapeutic agents, because they do not cause adverse effects, do not accumulate in the body, and are metabolized through specific proteases.

[0016] Thus, it is an object of the present invention to provide a composition based on a mixture of matricins [collagenocins (matricins obtained by heat treatment of native type I collagen of porcine origin) and elastocins (matricins obtained by heat treatment of bovine type I elastin)] and polymeric atelopeptide type I collagen with low molecular weight povidone.

[0017] Another object of the invention is to provide a composition:

[0018] (i) modulator of acute and chronic inflammation, hyperinflammation and cytokine release syndrome (ii) antifibrotic and fibrolytic

[0019] (iii) inducer of tissue regeneration

[0020] (iv) Inducer of peripheral immune tolerance based on a mixture of matricins (collagenocins, elastocins) with polymeric type I atelopeptide collagen which does not present rejection or have moderate or severe adverse side effects and which is applicable in the pharmaceutical, therapeutic and / or cosmetic area.

[0021] It is yet another object of the invention to satisfactorily alleviate and mitigate a) acute and chronic inflammatory conditions and systemic hyperinflammatory syndromes, including sepsis, secondary hemophagocytic lymphohistiocytosis, macrophage hyperactivation, acute respiratory failure, rotator cuff tendinitis, epicondylitis, bursitis, tenosynovitis, tendinitis, degenerative tendinosis, partial tendon rupture, rheumatoid arthritis (RA), scleroderma (SSc); b) scarring, keloid or hypertrophic scarring, diabetic ulcers, pressure ulcers, fracture consolidation; c) fibrosis such as pulmonary fibrosis, chronic obstructive pulmonary emphysema, hepatic cirrhosis, valvular heart disease, tendon sclerosis, achalasia and other similar conditions, which have as a common denominator the presence of acute or chronic inflammation and fibrosis.

[0022] Another object of the invention is to provide a chemical composition based on a mixture of matricins (collagenocins and elastocins) with polymeric type I atelopeptide collagen with povidone in different proportions [volume to volume (vol:vol), matricins: 0.1-9.9 with polymeric type I atelopeptide collagen: 9.9-0.1) of each of its components, which regulates inflammation and fibrosis in different pathologies as specified in the detailed description and in the examples cited below.

[0023] It is also an object of the invention to provide a composition based on a mixture of collagen matrixes, elastocins with polymeric type I atelopeptide collagen, administerable by different routes including topical, subcutaneous, intra-articular, intramuscular, oral, nasal and aerosol.

[0024] It is a further object of the present invention to provide a composition based on a mixture of collagenocins, elastocins with polymeric type I atelopeptide collagen with inflammation-modulating, antifibrotic, fibrolytic and tissue regeneration-inducing activity to compete advantageously with compositions of the compounds separately.

[0025] Consequently, the scope of the invention is determined in the area of ​​products applied to the pharmaceutical, therapeutic and / or cosmetic area.

[0026] The extracellular matrix (ECM) is the main component of any tissue and defines its structure and function. Under physiological and pathological conditions (alterations or loss of continuity in the ECM), processes such as migration, proliferation, cell differentiation, embryonic development, organogenesis, angiogenesis, cartilage remodeling, bone growth, tissue repair, wound healing, inflammation, growth, tumor metastasis, etc., depend on growth factors and cytokines contained in the ECM, as well as on ECM molecules that possess sequences similar to cytokines or growth factors and on soluble mediators synthesized de novo by parenchymal cells and the immune system.

[0027] Some of the modified PMECs have been shown to play a role in regulating chronic inflammation, and are currently being evaluated as part of the development of therapeutic strategies in cancer, neurodegenerative diseases, and inflammatory pathologies such as rheumatoid arthritis (RA) and osteoarthritis (OA).

[0028] Components of the extracellular matrix

[0029] The extracellular matrix (ECM) is a complex and dynamic structure that in mammals is composed of at least 1100 different proteins. Collectively, these extracellular proteins (ECPs) are known as the matrisosome. ECPs are classified into the collagen family, glycosaminoglycans (GAGs), proteoglycans, and glycoproteins. Matricryptins / matricins / matrixins

[0030] Matricins are small, bioactive peptides or cryptic subdomains with repetitive sequences, derived from the chemical, physical, or proteolytic degradation of extracellular matrix proteins (ECPs). The resulting peptides can have diverse shapes and sizes, and their biological activity differs from that of the original protein. They contain sequences with cytokine, chemokine, or growth factor activity. Matricins use integrins, growth factor receptors, cytokine receptors, or chemokine receptors as receptors. The binding of matricins to their ligand regulates proliferation, migration, cytokine production, protease production, ECP production, or apoptosis. There are two classes of matricins: "natural" matricins, which have exposed domains and generate direct signals in the extracellular matrix, and "cryptic" matricins, which require proteolytic, physical, or chemical processing to expose the ligand.Unlike traditional soluble growth factors, most matricins have low affinity for their receptors (micromolar concentrations) and have multiple valencies to increase avidity.

[0031] Elastin peptides, specifically those with the sequence X-Gli-XX-Pro-Gli, regulate chemotaxis, proliferation, and protease release in various cell lineages, such as fibroblasts and endothelial cells. They also induce angiogenesis and tubulogenesis, consequently having a direct effect on tissue repair and tumor metastasis. Their receptor, S-gal / EBP (spliced ​​[33-galactosidase / elastin binding protein], is expressed in fibroblasts, smooth muscle cells, endothelial cells, macrophages, neutrophils, leukocytes, monocytes, lymphocytes, and tumor cells.

[0032] Collagen peptides have several adhesion receptors such as elastin-binding protein (67 kDa), L-selectin, integrins, and chemokine receptors (CXCR1 and CXCR2). Their physiological functions include anti-angiogenesis, inhibition of cell proliferation and metastasis in melanoma, and inhibition of nerve migration and regeneration.

[0033] Clinical and therapeutic applications of matricryptins / matrixins / matricins

[0034] Alterations in body fluids regarding matricryptin levels have been useful as diagnostic, monitoring, or prognostic markers. For example, the concentration of endostatin, tumstatin, and endorepelin in the serum of patients with breast or lung cancer, idiopathic pulmonary arterial hypertension, in the bronchoalveolar lavage fluid of patients with pulmonary sarcoidosis, and in the cerebrospinal fluid of patients with neurodegenerative diseases has been found to be higher than in healthy individuals. Furthermore, antibodies generated against these neoepitopes have been used to develop solid-phase binding assays and to determine their concentration in the serum of patients with hepatic fibrosis. Modified matricryptins

[0035] Matricins are potential drugs, as demonstrated in several in vitro and in vivo trials in experimental models of various diseases. Among them is the 20 kDa matricin derived from the proteolysis of the globular domain of type XVIII collagen, endostatin. This matricin has anti-angiogenic properties when it binds to its endothelial receptors, α5pl and nucleolin, as it inhibits the FAK / c-Raf / MAP2K / p38 / ERK1 and CK2 (casein kinase 2) and CKD1 (cyclin-dependent kinase 1) pathways, respectively, and decreases the expression of mRNA for basic fibroblast growth factor (bFGF) and VEGF, thus inhibiting tumor growth and metastasis. The addition of an extra sequence at the N-terminus of endostatin improves its solubility and stability.This modified endostatin, called Endostar, received approval from the State Food and Drug Administration of China in 2005 for the treatment of patients with non-small cell lung carcinoma and is currently being evaluated for use in advanced gastric cancer, showing high effectiveness, good tolerability and few toxic effects.

[0036] Other studies have evaluated the combination of matricryptin with other drugs.

[0037] The 27 kDa matricin, derived from the proteolysis of the non-collagenous region (NC1) of the δ3 chain of type IV collagen, or tumstatin, binds to the integrin avp3 expressed on proliferating endothelial cells, inhibiting angiogenesis and, consequently, tumor growth. The binding of matricin to its receptor regulates the FAK / AkT / mT0R / 4E-BP1 pathway. Tumstatin, in combination with an anti-VEGF antibody, enhances its antitumor activity, as observed in a murine model of adenocarcinoma.

[0038] Other matriceptins require further evaluation in experimental models and clinical trials. For example, endorepelin, an 82 kDa matriceptin (V domain in the C-terminal region of the perlecan proteoglycan), has been shown to be neuroprotective, promoting nerve tissue repair through neurogenesis, neuroblast migration, and neuronal synapses after stroke. Endorepelin binds to the α5pl receptor, inducing ERK phosphorylation, which leads to the subsequent activation and stabilization of eIF4E and the inhibition of HIF-α activity, increasing VEGFR and VEGF expression and consequently neovascularization and inflammation. Furthermore, this matriceptin may be used as a therapy to inhibit p-amyloid deposition in Alzheimer's disease, reducing toxicity and restoring angiogenesis.

[0039] Polymeric type I atelopeptide collagen

[0040] Polymeric atelopeptide type I collagen is a radiation-irradiated mixture of porcine atelopeptide (pepsinized) type I collagen and polyvinylpyrrolidone (PVP) in a citrate buffer solution, which stabilizes the pH. Under culture conditions at 37 °C and neutral pH, the molecule does not form a gel, unlike collagen, and its electrophoretic, physicochemical, and pharmacological properties are modified by the covalent bond between the protein and the PVP. It has been demonstrated, both in vitro and in vivo, that the compound behaves differently than its components (collagen and PVP) separately.The addition of 1% polymeric atelopeptide type I collagen to synovial tissue cultures from patients with RA or OA has been shown to induce the negative regulation of the production of pro-inflammatory cytokines (IL-1 p, TNF-a, IL-8), the expression of adhesion molecules, ELAM-1 (endothelial leukocyte adhesion molecule), VCAM-1 (vascular cell adhesion molecule) and ICAM-1 (intercellular adhesion molecule), of the cyclooxygenase (Cox)-l enzyme, as well as collagenolytic activity through the modulation of the transcription factor NF-kB. Furthermore, polymeric type I collagen with PVP has been shown to induce the upregulation of the tissue inhibitor of MMPs, TIMP-1, the production of IL-10, the presence of regulatory T cells, and the proliferation of chondrocytes in co-cultures of synovial tissue and articular cartilage from patients with OA.However, the biopolymer appears to selectively stimulate synovial cell death through apoptosis. Finally, in vitro studies have shown that polymeric type I collagen with PVP stimulates the remodeling of the extracellular matrix (ECM) of the joint cavity, restoring the proportions of type I and III collagens without altering the total collagen content in the synovial tissue. It has also been observed to induce the recovery of type II collagen, cartilage oligomeric protein (COMP), and highly sulfated proteoglycans in cartilage.

[0041] Polymeric type I collagen with PVP, administered intra-articularly in a rat model of short- and long-term osteoarthritis (OA), has demonstrated that the biopolymer is a safe and effective treatment that modifies OA progression. It stimulates the upregulation of type II collagen and proteoglycans, the downregulation of type I collagen and MMP-13, and consequently acts as a chondroprotective agent and inducer of hyaline cartilage regeneration. The only adverse effect is pain at the injection site, lasting less than 15 minutes. The safety and efficacy of intra-articular administration of polymeric type I collagen with PVP were evaluated in two prospective, double-blind, placebo-controlled studies.The primary improvement criteria included the WOMAC functional disability questionnaire, the Lequesne index, pain intensity assessed by a visual analog scale (VAS), and the reduction in the number of nonsteroidal anti-inflammatory drug (NSAID) tablets. Secondary improvement criteria included disease status assessment (Likert scale) and evaluation of the molecule's effectiveness. Clinical improvement was defined as a 20 mm decrease in pain on the VAS and a final assessment showing at least a 20% improvement from baseline. Type II collagen degradation peptide (CTXII) concentrations were quantified in urine by ELISA, and the results were normalized to creatinine concentration. Laboratory variables assessed included complete blood count (CBC), liver function tests (LFTs), and urinalysis.The use of the biopolymer was found to induce a statistically significant improvement in the primary and secondary endpoints (p<0.05) in patients compared to baseline measurements and the placebo group. No alterations were found in blood cell count, liver function tests, or urinalysis. Furthermore, an increase in CTXII peptide concentration was observed in the placebo group compared to the group treated with polymeric type I collagen with PVP at 12 months of the study. Adverse reactions included pain at the injection site for 12 to 24 hours, and chemical arthritis developed in 2 patients (7%). These findings suggest that polymeric type I atelopeptide collagen has an excellent safety and clinical efficacy profile when administered intra-articularly.Thus, the biological role of polymeric type I atelopeptide collagen is not limited solely to providing protection, lubrication, and mechanical stability to the collagen network, cells in tissues, and joint surfaces with constant friction, but also induces chondrocyte proliferation, positively regulates the expression and synthesis of highly sulfated proteoglycans, type II collagen, cartilage oligomeric protein, and IL-10, and negatively regulates MMP-13 and pro-inflammatory cytokines, particularly IL-1p and TNF-α, acting as a disease progression modifier.

[0042] Brief Description of the Figures

[0043] Figure 1 shows the treatment of synovial tissue and cartilage co-cultures from patients with osteoarthritis (OA) or rheumatoid arthritis (RA) with a mixture of matricins and polymeric atelopeptide type I collagen (volume:volume = 1:1). The mixture of matricins and polymeric atelopeptide type I collagen (mixture, iv) restores proteoglycan concentration in the articular cartilage of patients with OA or RA (in E of Figure 1). In rows A and B of the Figure 1 micrograph: synovial tissue from patients with OA or RA. In rows C and D of the Figure 1 micrograph: cartilage from patients with OA or RA. Micrograph columns: Different treatments evaluated on day 7 of culture.

[0044] Figure 2 (B) illustrates how the mixture of matricins with polymeric type I atelopeptide collagen (mixture) increases chondrocyte proliferation (cell regeneration) in osteoarthritis (OA).

[0045] Figure 3 (A) shows the negative regulatory effect of the matrix mixture with polymeric type I atelopeptide collagen (iv) on TNF-α expression by synovial tissue cells of patients with rheumatoid arthritis (RA) or osteoarthritis (OA).

[0046] Figure 4 illustrates the negative regulation exerted by the mixture of matricins with polymeric type I atelopeptide collagen (iv) on IL-lp expression evaluated in cartilage and synovial culture supernatants from patients with rheumatoid arthritis (RA) or osteoarthritis (OA).

[0047] Figure 5 illustrates the downregulation of TNF-oc expression in the supernatants of cartilage and synovial co-cultures from patients with rheumatoid arthritis (RA) or osteoarthritis (OA) treated with the mixture of matricins and polymerized collagen (iv).

[0048] Figure 6 illustrates the downregulation of IL-8 expression in the supernatants of cartilage and synovium co-cultures from patients with rheumatoid arthritis (RA) or osteoarthritis (OA) treated with the mixture of matricins and polymerized collagen (iv).

[0049] Figure 7 illustrates the upregulation of IL-10 expression in the supernatants of cartilage and synovial tissue co-cultures from patients with rheumatoid arthritis (RA) or osteoarthritis (OA) treated with the mixture of matricins and polymerized collagen (iv).

[0050] Figure 8 illustrates the effect of the mixture of matricins and polymeric atelopeptide type I collagen (iv) on TIMP-1 expression in the supernatants of cartilage and synovial co-cultures from patients with rheumatoid arthritis (RA) or osteoarthritis (OA).

[0051] Figure 9 (B) illustrates the effect of the matrix and polymeric atelopeptide type I collagen mixture (iv) on the downregulation of TNF-α and IL-23 gene expression and the upregulation of Foxp-3 in cartilage and synovial co-culture tissues from patients with rheumatoid arthritis (RA). Figure 10 (A) and (B) show the effect of the matrix and polymeric atelopeptide type I collagen mixture (iv) on the downregulation of TNF-α gene expression and the upregulation of Foxp-3 in cartilage and synovial co-culture tissues from patients with osteoarthritis (OA).

[0052] Figure 11 (A), (B), (C), and (D) show the preventive and therapeutic effect of the matrix mixture with polymeric type I atelopeptide collagen in a collagen-induced arthritis (CIA) model in male DBAl / OlaHsd strain mice, preventing the increase in paw thickness and decreasing the arthritis severity score.

[0053] Figure 12 shows the leg thickness and histological image of joint sections stained with hematoxylin and eosin (100X), highlighting the negative regulatory effect of the matricin mixture with polymeric type I atelopeptide collagen (mixture) on inflammation and tissue damage. (A) in Figure 12 shows the first sacrifice, corresponding to 7 days after the last application of the mixture. (B) in Figure 12 shows the second sacrifice, corresponding to 56 days after the last application of the mixture.

[0054] Figure 13 (A), (B), (C) and (D) show the negative regulatory effect of the matrix mixture with polymeric type I atelopeptide collagen on the percentage of circulating effector CD4+ T cell subpopulations (Thl7 and Thl) and the positive regulatory effect on Foxp3+ regulatory T cells (Tregs) in the early arthritis model (preventive (A) and (B) of Figure 13) and in long-term (late or palliative) arthritis (C) and (D) of Figure 13.

[0055] Figure 14 (A), (B), (C) and (D) illustrate the effect of the mixture of matricins with polymeric type I atelopeptide collagen on the positive regulation of the NF-KB / IKB-Q complex of splenocytes in early arthritis (preventive (A) and (B) of Figure 14) and long-term arthritis (late or palliative (C) and (D) of Figure 14).

[0056] Figure 15 (A), (B), (C), (D) shows the beneficial effect of the matricin mixture with polymeric type I atelopeptide collagen (vol:vol) on weight, leg thickness, and temperature in rats with early and long-standing osteoarthritis (OA).

[0057] In (A), (B) of Figure 16, it is illustrated how the mixture of matricins with polymeric type I atelopeptide collagen (vol : vol ) preserves the radiographic 16 (A) and macroscopic 16 (B) characteristics of the joint in rats with early and long-standing osteoarthritis (OA).

[0058] Figure 17 shows how treatment with the matrix mixture with polymeric type I atelopeptide collagen (vol : vol ) preserves the proteoglycan content in the mid-zone of cartilage in rats with early and long-standing osteoarthritis (OA).

[0059] Figure 18 (A) and (B) illustrate how the mixture of matricins with polymeric type I atelopeptide collagen (vol : vol ) prevents fibrocartilage formation in rats with osteoarthritis (OA) by increasing the expression of type II collagen.

[0060] Figure 19 (A) and (B) illustrate how the mixture of matricins with polymeric type I atelopeptide collagen (vol : vol ) prevents fibrocartilage formation in rats with osteoarthritis (OA) by decreasing type I collagen production.

[0061] Figure 20 (A) and (B) show how the mixture of matricins with polymeric type I atelopeptide collagen (vol : vol ) prevents cartilage destruction by decreasing the expression of type 13 metalloproteinase (MMP-13) by chondrocytes.

[0062] The preceding descriptions exemplify how the object of the invention has a superior effect on inducing the synthesis of proteoglycans and type II collagen, on chondrocyte proliferation, on preserving the integrity of the joint cavity and cartilage, and on increasing the number of regulatory CD4+ T cells compared to the prior art. This also includes a decrease in some inflammatory mediators (IL-8, TNF-α), Thl7 cells, type I collagen, and MMP-13.

[0063] Detailed description of the invention

[0064] The present invention relates to a chemical composition modulating inflammation, inducing peripheral immune tolerance, anti-fibrotic, fibrolytic and inducing tissue regeneration based on a mixture of matricins [collagenocins (0.15-0.2% protein nitrogen), elastocins (1.5-2.0% protein nitrogen)] and polymeric type I atelopeptide collagen applicable in the pharmaceutical, therapeutic and / or cosmetic area.

[0065] The present invention relates to the study of the heat effect of protein derivatives of collagen (collagenocins) and elastin (elastocins) and their mixture in different proportions with polymeric type I atelopeptide collagen.

[0066] The present invention relates to a chemical composition that modulates inflammation, induces peripheral immune tolerance mechanisms, is antifibrotic and fibrolytic, and induces tissue regeneration, based on a mixture of collagens, elastokines, and polymeric type I atelopeptide collagen, applicable in the pharmaceutical, therapeutic, and / or cosmetic fields. The composition of the present invention includes collagen and elastin, combining a + b, where:

[0067] a) Collagen and elastin matrices. Compound based on:

[0068] I. an aqueous solution of 0.9-1.1% w / w of porcine type I collagen from skin, native, atelopeptide and highly purified.

[0069] II. a non-viscous aqueous solution of 5-10% elastin hydrolysates.

[0070] Matricins are obtained by thermohydrolysis in a pre-sterilized, moist-heat mixing kettle running at 60 ± 5 rpm. A portion of porcine type I collagen is mixed in citrate buffer (I), and water-soluble bovine type I elastin obtained by enzymatic hydrolysis (II) is gradually added. The mixture is stirred and transferred to a previously sterilized flask, which is then covered with medical-grade paper. This flask is subjected to an autoclave cycle for 30 minutes at 105–125 °C and a pressure of 13–25 psi. The solution is filtered through a membrane to maintain sterility. The solution is then subjected to a second autoclave cycle as a standard sterilization procedure. Collagen hydrolysates range in molecular weight from 1000 to 25,000 Da, while elastin hydrolysates range from 200 to 150,000 Da.The total particle size of the hydrolysates is considered to be preferably between 5,000 and 150,000 Da, and the nitrogen content in the range of 4.4–7.2 mg / ml. The collagen and elastin used have a degree of purification greater than 90%, which implies that there are no impurities responsible for the desired therapeutic effect.

[0071] With :

[0072] b) Polymeric type I atelopeptide collagen. The porcine type I atelopeptide collagen of the present invention, with a purity exceeding 90%, is dissolved in a buffered solution with a slightly acidic pH and mixed with a vinyl polymer. The vinyl polymer is selected from a group consisting of polyvinyl alcohol, polyvinyl chloride, polyvinylpyrrolidone, or any other pharmacologically acceptable vinyl polymer. This mixture is irradiated with gamma rays, obtaining controlled crosslinking between the collagen and the vinyl polymer. The resulting composition is preferably sterilized using standard procedures appropriate for heat-labile materials. It is considered that the composition of the present invention can be adequately formulated using collagen in the range of 5 to 30 mg / ml and the vinyl polymer from 5 to 50% of the composition.However, other wider intervals are possible, still effective, although less so, and still higher than those of alternative products.

[0073] It is clear that the present invention does not refer to a biopharmaceutical in which the amino acids have been sequenced, but to avoiding contamination with proteins other than those mentioned that are responsible for the desired therapeutic effect.

[0074] Once the two parts of the mixture are obtained, they are combined in different proportions ranging from 0.1 to 9.9 ml for the matricins with 9.9 to 0.1 ml of polymeric type I atelopeptide collagen, which have been shown to have different activities, including inducing healing, increasing the speed of healing, high-quality repair and tissue regeneration, as well as negatively regulating inflammation and inducing peripheral immune tolerance.

[0075] The mixture of matricins (collagenacins and elastocins) with polymeric type I atelopeptide collagen (in proportions of 1:9, 3:7 and 5:5) has antifibrotic, fibrolytic, inflammation-modulating, peripheral immune tolerance-inducing, and tissue-regenerating action in chronic degenerative joint diseases and may be indicated for the treatment of rotator cuff tendinosis, epicondylitis, bursitis, tenosynovitis, tendinitis, degenerative tendinosis, partial tendon rupture, scleroderma, scarring, keloid or hypertrophic scarring, pulmonary fibrosis, chronic obstructive pulmonary emphysema, hepatic cirrhosis, valvular heart disease, diabetic ulcers, pressure ulcers, tendon sclerosis, fracture consolidation, achalasia and other similar medical conditions.which have as a common denominator the presence of chronic inflammation and fibrosis through various mechanisms that are detailed later and were carefully studied.

[0076] The dosage and dosing regimen are determined by the progression of the condition and the route of administration. It can be administered intravenously to open lesions when the aim is to induce healing and prevent fibrosis, or via local injection, subcutaneous injection, intra-articular injection, intramuscular injection, oral injection, nasal injection, aerosol application, patch application, and other methods when treating a chronic local or systemic inflammatory and / or fibrosing condition. It is worth noting that, to date, no adverse effects of the composition are known, which implies that the composition of the invention is harmless. This conclusion is based on in vitro and in vivo studies evaluating the effect of the matricin mixture (collagenocins and elastocins) with polymeric type I atelopeptide collagen on inflammation and extracellular matrix (ECM) turnover of cartilage and the synovial membrane in patients with osteoarthritis (OA) or rheumatoid arthritis (RA), as well as in murine and rat models.

[0077] Example 1

[0078] Evaluation of the effect of the mixture of matricins (collagenocins and elastocins) and polymeric type I atelopeptide collagen in co-cultures of synovial membrane and cartilage from patients with rheumatoid arthritis (RA) and osteoarthritis (OA).

[0079] Osteoarthritis (OA) ranks first among the 10 most frequent causes of morbidity in people aged 65 and over, with a prevalence of 2.3% (1.7–2.9%), while rheumatoid arthritis (RA) has a prevalence of 0.3% (0.1–0.6%) in the Mexican population. There is no specific treatment for OA, and in the case of RA, most of the drugs used produce significant side effects, while their cost is very high. With the intention of finding more efficient therapeutic options than those currently available, and with fewer adverse effects for both chronic inflammatory diseases, the effects of a mixture of modified PMECs (matricines with polymeric atelopeptide type I collagen) were evaluated.

[0080] For this purpose, co-cultures of cartilage and synovium were performed from 9 patients with RA and 8 patients with OA undergoing hip or knee prosthesis replacement (Table 1). The articular tissue was cultured with (i) RPMI culture medium (control), (ii) 1% matricins (collagenocins and elastocins), (iii) 1% polymeric atelopeptide type I collagen (polymeric collagen), and (iv) a mixture of 1% matricins plus polymeric atelopeptide type I collagen (mixture), for 7 days. Supernatants and tissues from days 1 and 7 were collected to determine the effect of different conditions (ii, iii and iv) on ECM turnover, cell proliferation (ki 67) and on the concentration of soluble mediators associated with inflammation (IL-1, IL-8, IL-10, IL-12, TNF-oc and IFN-y).

[0081] Table 1. Demographic data and clinical history of the disease

[0082]

[0083]

[0084] 1 Gender: female, M: male, 2 D.E.: standard deviation; 3 DMARDs: Disease-Modifying Drugs; 4 NSAIDs: Non-Steroidal Anti-Inflammatory Drugs; 5 ESR: Erythrocyte sedimentation rate.

[0085] Determination of proteoglycans in co-cultures of cartilage and synovial tissue from patients with RA or OA.

[0086] The mixture (condition iv) had a greater and statistically significant effect on inducing tissue remodeling than the culture conditions with matricin (ii), polymerized collagen (iii), and the control (i) because it increased the concentration of highly sulfated proteoglycans (blue-stained area, Fig. 1A-1D). It should be noted that inflammatory foci decreased in the synovial membranes treated under all three culture conditions (ii), (iii), and (iv), with the main effect observed in tissues treated with mixture (iv) on day 7 of culture. In samples from OA patients treated with polymerized collagen (iii), the relative percentage of proteoglycans increased 4.0 times, and in those treated with mixture (iv) 5.1 times, on day 7 of culture. the control culture (In E of Figure 1). The treatment with the mixture (iv) was 1.1 times higher than the treatment with polymerized collagen (iii).

[0087] Figure 1. Treatment with the mixture of matricins and polymeric atelopeptide type I collagen (IV) restores proteoglycan concentration in articular cartilage from patients with osteoarthritis (OA) or rheumatoid arthritis (RA). Rows A and B: Synovial tissue from patients with OA and RA. Rows C and D: Cartilage from patients with OA and RA. Columns: Control 0: Tissue under basal conditions. Control 7: Control tissue cultured for 7 days and untreated. Matricins: Mixture of collagens and elastokines. Polymerized atelopeptide type I collagen (polymerized collagen). Mixture: Matricins and polymerized collagen. Highly sulfated proteoglycans stain blue. (E) Relative percentage of proteoglycan content. Basal: tissue under basal conditions. Control Day 7: control tissue cultured with RPMI for 7 days and which received no treatment.Matricins: mixture of collagens and elastocins. Mixture: mixture of matricins and polymeric atelopeptide type I collagen. Each culture was performed in triplicate for each condition, of which at least two sections of each tissue were analyzed. The results are expressed as the mean (thick bar) and median (thin bar) of the percentage of proteoglycans in 9 co-cultures from patients with RA and 8 with OA. *R<0.01.

[0088] Chondroproliferation assessment: The addition of matricins (ii), polymerized collagen (iii), or mixture (iv) significantly decreased the proliferation (Ki 67) of synovial membrane cells from RA patients, but not of OA cells (In A of Fig. 2 top panel) vs. the control culture. In contrast, cartilage chondrocyte proliferation from OA patients treated with mixture (iv) increased 10–12 times vs. The control (7.3 + 3.3 vs. 1.2 + 0.5 or 0.3 + 0.3, treated cultures vs. basal or untreated culture; F<0.05; In (B) of Fig. 2, lower panel). While the increase was between 5 and 6 times in the cultures treated with polymerized collagen (iii) vs. the controls. No cell death was determined by cytotoxicity of the compounds, the latter being evaluated by histology and by the presence of the Ki-67 antigen. Figure 2.The mixture of matricins with polymeric atelopeptide type I collagen (mixture) increases chondrocyte cell proliferation (cell regeneration) in osteoarthritis. Percentage of (A) synovial cells and (B) proliferating cartilage. Rheumatoid arthritis (RA): left panel. Osteoarthritis (OA): right panel. Basal: tissue under basal conditions. Day 7 Control: control tissue cultured with RPMI for 7 days. Matricins: collagen and elastokine matrices. Mixture: matricins and polymeric atelopeptide type I collagen. Each culture was performed in triplicate for each condition, with at least two sections of each tissue analyzed. Results are expressed as the mean ± standard error (SEM) of the percentage of immunoreactive cells in 9 co-cultures from patients with RA and 8 with OA. *P<0.05.

[0089] Negative regulation of pro-inflammatory cytokine synthesis by cartilage and synovial cells in patients with RA or OA.

[0090] The addition of both matricins (ii) and polymerized collagen (iii) and the mixture of matricins with polymerized collagen (iv) to the co-cultures of AR or OA decreased tissue expression of TNF-oc vs. controls in synovial tissue (A and B of Fig. 3).

[0091] Figure 3. Negative regulatory effect of the mixture of matricins with polymeric atelopeptide type I collagen (iv) on TNF-α expression. TNF-κB producing cells in (A) of Figure 3, synovial tissue, or in (B) of Figure 3, cartilage from co-cultures. Rheumatoid arthritis (RA): left panel. Osteoarthritis (OA): right panel. Basal: tissue under basal conditions. Day 7 control: tissue cultured with RPMI for 7 days and untreated. Matricins: mixture of collagenins and elastokinins. Mixture: mixture of matricins with polymeric atelopeptide type I collagen. Each culture was performed in triplicate for each condition, and at least two sections of each tissue were analyzed. Results are expressed as the mean ± standard error (SEM) of the percentage of immunoreactive cells in 9 co-cultures from patients with RA and 8 with OA. *P< 0.002.

[0092] Negative regulation of cytokine release into the supernatant of cartilage and synovium co-culture from patients with RA or OA.

[0093] The addition of matricins (ii), polymerized collagen (iii) and the mixture of matricins and polymerized collagen (iv) to AR or OA co-cultures decreased the expression of IL-1 and TNF-oc vs. controls in the supernatants on day 7 of culture (Fig. 4 and Fig. 5, respectively).

[0094] Figure 4. Negative regulation of IL-ip expression in the supernatants of cartilage and synovial co-cultures from patients with rheumatoid arthritis (RA) or osteoarthritis (OA) treated with the mixture of matricins and polymerized collagen (iv). Data represent the mean ± SD of 9 cartilage and synovial tissue co-cultures from patients with RA and 8 co-cultures from patients with OA. RA: left panel. OA: right panel. Basal: tissue under basal conditions. Day 7 Control: control tissue cultured with RPMI for 7 days. Matricins: collagenokines and elastokines. Mixture: mixture of matricins and polymerized atelopeptide type I collagen. *P < 0.008.

[0095] Figure 5. Negative regulation of TNF-α expression in the supernatants of cartilage and synovial co-cultures from patients with rheumatoid arthritis (RA) or osteoarthritis (OA) treated with the mixture of matricins and polymerized collagen(iv). Data represent the mean ± SD of 9 cartilage and synovial tissue co-cultures from patients with RA and 8 co-cultures from patients with OA. RA: left panel. OA: right panel. Baseline: tissue under basal conditions. Day 7 Control: control tissue cultured with RPMI for 7 days. Matricins: collagens and elastocins. Mixture: mixture of matricins and polymerized athelopeptide type I collagen. *P < 0. 001 .

[0096] A similar pattern was observed with respect to IL-8 concentration in the co-culture supernatants (Fig. 6). The greatest decrease was observed in co-cultures of tissues from RA patients treated with the matrix and polymerized collagen mixture (iv). Neither IL-12 nor IFN-γ were detected in the culture supernatants.

[0097] Figure 6. Negative regulation of IL-8 expression in the supernatants of cartilage and synovial co-cultures from patients with rheumatoid arthritis (RA) or osteoarthritis (OA) treated with the mixture of matricins and polymerized collagen (iv). Data represent the mean ± SD of 9 cartilage and synovial tissue co-cultures from patients with RA and 8 co-cultures from patients with OA. RA: left panel. OA: right panel. Basal: tissue under basal conditions. Day 7 Control: control tissue cultured with RPMI for 7 days. Matricins: collagenokines and elastokines. Mixture: mixture of matricins and polymerized athelopeptide type I collagen. *P<0.05 OO 6. Increase in the expression of anti-inflammatory mediators in the supernatants of cartilage and synovial co-cultures from patients with RA or OA

[0098] The addition of matricins (ii), polymerized collagen (iii), or the mixture of matricins and polymerized collagen (iv) to the RA or OA co-cultures increased the concentration of IL-10, a cytokine with anti-inflammatory activity, by 10 to 25 times in the supernatants of the 7-day co-cultures vs. on day 1 of culture (Fig. 7). None of the culture conditions had any effect on TIMP-1 levels (Fig. 8).

[0099] Figure 7. Upregulation of IL-10 expression in the supernatants of cartilage and synovial tissue co-cultures from patients with rheumatoid arthritis (RA) or osteoarthritis (OA) treated with the mixture of matricins and polymerized collagen (iv). Data represent the mean ± SD of 9 cartilage and synovial co-cultures from patients with RA and 8 co-cultures from patients with OA. Each culture was performed in triplicate for each condition. RA: left panel. OA: right panel. Basal: tissue under basal conditions. Control Day 7: control tissue cultured with RPMI for 7 days. Matricins: collagenins and elastokines. Mixture: mixture of matricins and polymerized atelopeptide type I collagen. *P < 0.006.

[0100] Figure 8. Effect of the mixture of matricins and polymeric atelopeptide type I collagen (iv) on TIMP-1 expression in the supernatants of cartilage and synovial co-cultures from patients with rheumatoid arthritis (RA) or osteoarthritis (OA). Data represent the mean ± SD of 9 cartilage and synovial co-cultures from patients with RA and 8 co-cultures from patients with OA. RA: left panel. OA: right panel. Basal: tissue under basal conditions. Day 7 Control: control tissue cultured with RPMI for 7 days. Matricins: collagens and elastokines. Mixture: matricins and polymeric atelopeptide type I collagen. *P < 0.006.

[0101] The mixture of matricins and polymeric atelopeptide type 1 collagen regulates the gene expression of pro- and anti-inflammatory cytokines in cartilage and synovial co-culture tissues from patients with RA or OA

[0102] Polymerized collagen (iii) and the mixture of matricins and polymerized collagen (iv) negatively regulated the transcription of tnf-α and il-23 (In A and B of Figure 9) and positively regulated that of foxp3 at statistically significant levels compared to the control (In (B) of Fig. 9). Figure 9. Effect of the mixture of matricins and polymeric atelopeptide type I collagen (iv) on the regulation of gene expression of cytokines and pro- and anti-inflammatory factors in the tissues of cartilage and synovial co-cultures from patients with rheumatoid arthritis (RA). Figure 9 (A) shows the quantification of gene expression of pro-inflammatory cytokines (il-lfl, tnf-a, il-6, 11-23) and Figure 9 (B) shows cytokines and anti-inflammatory factors (il-10, foxp3) in the synovial membrane and cartilage co-cultured for 7 days.The bars show the mean ± SD of transcript levels from samples normalized across the expression of the constitutive gadph gene as determined by 2AAct. Data represent the mean ± SD of 9 co-cultures of cartilage and synovium from RA patients. Each culture was performed in triplicate for each condition. Basal: tissue under basal conditions. Day 7 Control: control tissue cultured with RPMI for 7 days. Matricins: collagens and elastokines. Mixture: matricins and polymeric atelopeptide type I collagen. Whereas the addition of polymerized collagen (iii) or the mixture of matricins and polymerized collagen (iv) to joint tissue co-cultures from patients with OA induced a negative regulation of tnf-a gene expression and a positive regulation of il-6 and foxp3 at statistically significant levels with respect to the control (Fig. 10).

[0103] Figure 10. Effect of the mixture of matricins and polymeric atelopeptide type I collagen (iv) on the regulation of gene expression of pro- and anti-inflammatory cytokines and factors in co-cultured cartilage and synovial tissues from patients with Osteoarthritis (OA). (A) Quantification of gene expression of (A) pro-inflammatory cytokines (il-lfl, tnf-a, il-6, IL-23) and (B) anti-inflammatory cytokines and factors (11-10, foxp3) in co-cultured synovial membrane and cartilage for 7 days. Bars show the mean ± SD of transcript levels from samples normalized through the expression of the constitutive gadph gene determined by 2AAct. The data represent the mean ± SD of 9 co-cultures of cartilage and synovium from patients with RA. Each culture was performed in triplicate for each condition. Baseline: tissue under basal conditions. Control Day 7: control tissue cultured with RPMI for 7 days.Matricins: collagens and elastocins. Mixture: matricins and polymeric type I atelopeptide collagen.

[0104] It was shown that the addition of the matrix mixture with polymeric atelopeptide type I collagen (iv) to cartilage and synovial co-cultures from RA patients decreased synovial cell proliferation and the synthesis of pro-inflammatory cytokines such as: IL-ip, IL-8, IL-23 and TNF-oc, which could be directly or indirectly related to the observed increase in IL-10 levels and Foxp3 gene expression.

[0105] On the other hand, in the co-cultures of joint tissue from patients with OA treated with the mixture of matricins and polymeric type I atelopeptide collagen (iv), an increase in the proliferation of chondrocytes was determined, evaluated by the presence of the Ki-67 antigen (tissue regeneration), an increase in the recovery of sulfated proteoglycans (10-12X; recovery of cartilage PMECs), a decrease in pro-inflammatory mediators (TNF-oc) and an increase in the anti-inflammatory cytokine, IL-10, and the gene expression of Foxp3, a factor associated with regulatory T cells.

[0106] In both diseases, the synovial membrane, rather than the cartilage, appears to be the tissue that contributes most significantly to the synthesis of soluble pro-inflammatory factors. The action of the different conditions (ii, iii, and iv) seems to depend on the stage of cell activation, since the addition of matricins, polymeric atelopeptide collagen type I, or a mixture of both, to resting cells does not induce any response; that is, it has no effect.

[0107] The results obtained are consistent with the analysis of various PMECs that have been used as potentially therapeutic molecules in animal models of:

[0108] a) autoimmune diseases, such as RA in which synthetic fibronectin peptides have been evaluated to inhibit or interrupt the infiltration of inflammatory cells into the joint cavity, or thrombospondin-1 (TSP-1) and angiostatin (plasminogen fragments) as a strategy to inhibit angiogenesis.

[0109] b) Suppression or inhibition of tumor growth and metastasis, such as endostatin (collagen fragments XVII I) - angiostatin and soluble Tie-2 which increases the suppression of prosthetic tumor growth and melanoma or endostatin complexes with DNA within cationic liposomes to prevent metastasis of osteosarcoma.

[0110] In conclusion, the mixture of matricins and polymeric atelopeptide type I collagen (iv) induces tissue remodeling, promoting the recovery of cartilage proteoglycans, negatively regulating the expression of some pro-inflammatory cytokines, and stimulating chondrocyte proliferation and the production of anti-inflammatory cytokines. Example 2

[0111] Evaluation of the inflammation-regulating, immuno-tolerance-inducing effect and mechanism of action of the matricin mixture (collagenocins and elastocins) with polymeric type I atelopeptide collagen in a murine model of collagen-induced arthritis (CIA)

[0112] The objective of the study was to evaluate whether the intramuscular application of a mixture of matricins (elastocins and collagenocins) with polymeric type I atelopeptide collagen or the mixture combined with methotrexate (MTX) prevented the onset or progression of collagen injection-induced arthritis (CIA) in rodents. The effect was determined at the clinical, histological, and molecular levels by comparing the results with the gold standard treatment: MTX.

[0113] To achieve this, 3 models were implemented:

[0114] Toxicity Model. Twenty-one 8-week-old male DBAl / OlaHsd strain mice were treated with 100 / I of (i) Placebo (citrate buffer)

[0115] (ii) Polymeric type I atelopeptide collagen (polymerized collagen),

[0116] (iii) The mixture of matricins and polymeric type I atelopeptide collagen (matricins + polymerized collagen) (iv) MTX (2.5 mg / kg),

[0117] (v) Polymeric type I atelopeptide collagen and MTX (vol : vol ) (polymerized collagen + MTX) ,

[0118] (vi) The mixture of matricins + polymerized collagen and MTX (vol : vol ) (matricins + polymerized collagen + MTX), once a week for six weeks.

[0119] Early Arthritis Model. Fifty mice were immunized at the base of the tail with 100 g of emulsified chicken type II collagen in Freund's complete adjuvant. The booster was applied on day 21 at the base of the tail with 100 g of emulsified type II collagen in Freund's incomplete adjuvant. On the same day as the booster, intramuscular (i-vi) treatments were administered once a week for six weeks.

[0120] Established Arthritis Model. Arthritis was induced as in the previous model in 50 mice and treatments were applied intramuscularly, after 2 weeks of booster, every week for 6 weeks.

[0121] From each model and each treatment group, 3 animals were sacrificed at weeks 7 and 13 post-treatment. The spleen, lymph nodes, joint tissue, liver, lungs, kidneys, and heart were obtained in order to assess inflammatory infiltrate and predominant cell populations, cytokine expression, and ECM turnover.

[0122] Clinimorphometric measurements were performed under baseline conditions and weekly for 13 weeks. Leg thickness increase was measured using an electronic caliper. Weight and temperature were also recorded. The degree of inflammation was assessed in each leg according to the following scale: 0, no inflammation; 1, toe erythema (mild); 2, erythema, leg inflammation, and ankle edema (moderate); and 3, edema, ankylosis, and complete immobility of the limb (severe). The assessment was performed blindly by a single observer.

[0123] Histological evaluation. The inflammatory infiltrate and tissue architecture were evaluated by H&E and Masson's trichrome staining, and proteoglycan content was assessed using the PAS technique. Cytometric evaluation. The percentage of Thl cell populations (CD4) was determined by FACS. + / IFN-y +), Th2 (CD4+ / IL-4 +), Treg (CD4+ / FOXP3+) and Thl7 (CD4+ / IL-17 +) in spleen cells.

[0124] Effect of the mixture of matricins and polymeric type I ate lopeptide collagen in the toxicity model.

[0125] The anatomical, macroscopic, and histological analysis of the kidneys, heart, lungs, spleen, lymph nodes, and hind legs of the animals showed normal tissue structure. None of the treatments evaluated in the study produced toxicity in the mice.

[0126] Effect of the mixture of matricins with polymeric type I atelopeptide collagen on clinimetrics in the early (preventive) and long evolution (late or palliative) arthritis model.

[0127] DBAl / OlaHsd mice developed signs of arthritis 21 days after the second immunization. In the preventative model, severe arthritis was reached around days 35-42. Arthritic mice treated with polymerized collagen (ii), matricins + polymerized collagen (iii), polymerized collagen + MTX (v), or matricins + polymerized collagen + MTX (vi) showed significant reductions in arthritis severity compared with placebo (In A and B of Fig. 11).

[0128] In the long-term model, treatments with polymerized collagen (ii), matricin + polymerized collagen (iii), polymerized collagen + MTX (v), or matricin + polymerized collagen + MTX (vi), rather than methotrexate monotherapy, significantly suppressed arthritis severity scores (Figures C and D in Fig. 11). Notably, paw thickness was normal in these groups. Methotrexate treatment did not reduce edema and induced fever.

[0129] Figure 11. Preventive and therapeutic effects of the matricin mixture with polymeric type I atelopeptide collagen in mice with collagen-induced arthritis (CIA). Eight-week-old DBAl / OlaHsd strain male mice were treated intramuscularly with 100.1 of (i) placebo (citrate buffer), (ii) polymerized collagen, (iii) matricins + polymerized collagen, (iv) MTX (2.5 mg / kg), (v) polymerized collagen + MTX (vol:vol), (vi) matricins + polymerized collagen + MTX (vol:vol), once a week for 6 weeks. Leg thickness in the early arthritis model (A) of Figure 11 and the long-term arthritis model (C) of Figure 11. Arthritis score in the early arthritis model (B) of Figure 111 (n=6) and the long-term arthritis model (D) of Figure 111 (n=6). Data represent the mean ± SD (each group, n=6). *P<0.05.

[0130] Histopathological analysis of the effect of the mixture of matricins with polymeric type I atelopeptide collagen in the early (preventive) and long evolution (late or palliative) arthritis model.

[0131] Histopathological analysis was performed on the hind legs of mice with CIA. Representative images of H&E and PAS-stained sections from the groups treated with (i) placebo, (ii) polymerized collagen, (iii) matricins + polymerized collagen, (iv) MTX (2.5 mg / kg), (v) polymerized collagen + MTX (vol:vol), (vi) matricins + polymerized collagen + MTX (vol:vol) are presented in Figure 12. Mice with CIA exhibited extensive infiltration of inflammatory cells, synovial hyperplasia, loss of joint space, and bone erosion. Treatment with (ii) polymerized collagen reduced the degree of inflammation and preserved joint structure. While treatment with (v) polymerized collagen + MTX (vol : vol ) resulted in a significant reduction of inflammatory infiltrates and recovery of tissue architecture.However, treatment with the mixture of (iii) matricins + polymerized collagen, with or without (vi) MTX, resulted in the disappearance of cellular infiltration, inhibition of pannus formation, and inhibition of cartilage and bone destruction in arthritic joints in both models (preventive and palliative). MTX induced some tissue abnormalities, such as the presence of nodules (amorphous fibrin tissue) and poor-quality lesion repair tissue, as well as inflammatory infiltrates. The effect of the different conditions was maintained until the second sacrifice (Fig. 12). Figure 12. Palliative effect of the mixture of matricins with polymeric type I atelopeptide collagen on histological damage in mice with CIA. Hematoxylin and eosin stain. (In A of Figure 12) Representative section from the first sacrifice. (In B of Figure 12) Representative section of the second sacrifice. The magnification of the histologies corresponds to 100X.

[0132] Effect of the mixture of matricins with polymeric atelopeptide type I collagen in the early (preventive) and long-term (late or palliative) arthritis model on the percentage of CD4+ T cell subpopulations of the spleen. The percentage of autoimmune pro-inflammatory cells, Thl 7, in the spleen of CIA-free mice (controls) was ~1.3%, while in CIA-free mice in the early arthritis model it was 2.5% and 4.3%, for the first and second sacrifice, respectively (In A and B of Figure 13) and 2.4% and 3.6%, for the first and second sacrifice, respectively in the long-term arthritis model (In C and D of Figure 13).

[0133] In the treatment group with the mixture of (iii) matrices + polymerized collagen and (vi) matrices + polymerized collagen + MTX, a sustained and statistically significant reduction in Thl 7 cell levels was determined within intervals considered normal, both in the early arthritis model (first and second sacrifice: 1.0-1.4% and 0.9-1.35%; in A and B of Figure 13), and in long-standing arthritis (first and second sacrifice: 0.5-0.45% and 0.9-1.45%; in C and D of Figure 13). (iv) MTX was not as effective in suppressing the percentage of Thl 7 cells from the spleen.

[0134] The number of circulating Th2 cells increased with treatment of the mixture of (vi) matrices + polymerized collagen + MTX during the second sacrifice of palliative model mice vs. placebo-treated control group of CIA mice (In D of Figure 13).

[0135] Exclusively, in the long-standing arthritis model, the mixture of (iii) matricins + polymerized collagen or (vi) matricins + polymerized collagen + MTX decreased the percentage of Thl cells in the spleen to normal concentrations during the first and second sacrifice (In C and D of Figure 13), while (iv) MTX decreased the number of Thl cells in the spleen to statistically significant levels.

[0136] It is worth noting that treatment of early and long-standing arthritis with the mixture of (iii) matricins + polymerized collagen or (vi) matricins + polymerized collagen + MTX induced a significant increase in Foxp3+ regulatory T cells (Tregs) compared to that of arthritic mice treated with (i) placebo or vehicle and to that of healthy mice in both the first and second sacrifices (In A, B, C and D of Figure 13). In contrast, (iv) MTX induced a significant reduction in the percentage of these cells in both models.

[0137] Figure 13. Negative regulatory effect on the percentage of circulating effector CD4+ T cell subpopulations of the matrix mixture with polymeric type I atelopeptide collagen in the early (preventive) and long-standing (late or palliative) arthritis models. Spleen cells obtained ex vivo in the early arthritis model during the first sacrifice (Figure 13, A) and second sacrifice (Figure 13, B). Spleen cells obtained ex vivo in the long-standing arthritis model during the first sacrifice (Figure 13, C) and second sacrifice (Figure 13, D). Intracellular production of IL-17A, IL-4, IFN-γ, and Foxp3 was detected by flow cytometry. The results are representative of 6 mice analyzed in each group. The horizontal dotted line represents the average normal values, obtained from mice without CIA (n = 3). Data represent the mean ± standard error of the mean (SEM). *P < 0.05.

[0138] Negative regulatory effect of the mixture of matricins with polymeric atelopeptide type 1 collagen in the early (preventive) and long evolution (late or palliative) arthritis model on the transcription factor NF-kB.

[0139] We inferred that the mechanism of action of the matrix + polymerized collagen mixture could be mediated through the regulation of the transcription factors NF-κB and AP-1. In particular, NF-κB regulates the expression of proinflammatory cytokines, chemokines, cell adhesion molecules, and apoptosis. In light of this knowledge, the expression of NF-κBp65 and its inhibitor IKBCI was analyzed in splenocytes ex vivo (Fig. 14).

[0140] In mice with early or long-standing CIA, the percentage of NF-KBp65+ (active form) and IKB‰+ cells was higher, while that of NF-KB+ / IKB‰+ (inactive NF-KB) was lower compared to healthy mice and arthritic mice treated with the mixture of (iii) matricins + polymerized collagen or (vi) matricins + polymerized collagen + MTX (Fig. 14). In short- and long-standing arthritic mice, the percentage of NF-KB+ / IKB‰+ complexes increased significantly in those groups treated with (ii) polymerized collagen; (vi) the mixture of matricins + polymerized collagen + MTX; and (iii) the mixture of matricins + polymerized collagen, suggesting downregulation of the transcription factor and consequently of inflammation.

[0141] Figure 14. Effect of the matrix mixture with polymeric atelopeptide type I collagen on the upregulation of the NF-KB / IKB-OI complex in splenocytes from early (preventive) and long-standing (late or palliative) arthritis. Spleen cells from early arthritis model mice during the first (in A of Figure 14) and second (in B of Figure 14) sacrifice. Spleen cells from long-standing arthritis model mice during the first (in C of Figure 14) and second (in D of Figure 14) sacrifice. Intracellular levels of NF-KBp65 and IKBQ were detected by flow cytometry. The results are representative of 6 mice analyzed in each group. The horizontal dashed line represents the average normal values ​​obtained from mice (n = 3) without CIA. The data represent the mean ± SEM. *P <0.05.

[0142] Adverse effects.

[0143] After the placebo injection or the different treatments throughout the study, the only adverse event observed was pain at the injection site, which lasted less than 15 minutes. In conclusion, the toxicity model showed that there was no damage to the organs analyzed, in any of the conditions evaluated.

[0144] The study demonstrated that both the mixture of (iii) matrices + polymerized collagen as monotherapy, as well as the mixture of (vi) matrices + polymerized collagen + MTX, exhibit both preventive and palliative effects in the short- and long-term CIA model, through the negative regulation of the Thl7 cell subpopulation (1.5-2.0 and 3.0-4.0X, respectively) and the positive regulation of Treg cells (5.0-6.0 and 5.5-6.0, respectively). The mechanism of action appears to be directly related to the regulation of the transcription factor NF-α.

[0145] Clinimorphic analysis showed that the best treatment was the mixture of (iii) matricins + polymerized collagen, followed by (vi) matricins + polymerized collagen + MTX, then (ii) polymerized collagen, and finally (v) polymerized collagen + MTX, with MTX being the least effective treatment (P<0.05). The joints of mice with ASD treated with the (iii) mixture of matricins + polymerized collagen maintained a similar architecture to normal, without inflammatory infiltrates, bone erosions, or loss of joint space, as well as proteoglycan content. The PMEC-based treatments were safe and effective. No adverse effects were reported. Example 3

[0146] Evaluation of the effect of the mixture of matricins (collagenocins and elastocins) with polymeric atelopeptide type I collagen on articular cartilage repair and inflammation regulation in a rat model of osteoarthritis (OA).

[0147] The objective of the study was to evaluate whether the intra-articular application of a mixture of matricin and polymerized collagen in different proportions could modify the progression of early and long-standing osteoarthritis in a rat model. The effect was determined at the clinical, radiological, histological, and molecular levels.

[0148] For this purpose, 48 male Wistar rats of 130-150 g, grown under pathogen-free conditions, were analyzed, which were divided into 2 control groups and 3 models (In A, B and C of Fig.

[0149] 15) :

[0150] Control Group without Surgery: Six rats without meniscectomy, surgery, training, or treatment were raised ad libi tum. Three animals were sacrificed at the ninth and three at the fifteenth week after the start of the study.

[0151] Control Group with Surgery: Six rats without meniscectomy, but with surgery, with 3 weeks of training and no treatment were raised ad libitum. Three animals were sacrificed in the ninth and three in the fifteenth week after the start of the study. Toxicity Model: Six rats without meniscectomy, without surgery, but subjected to 3 weeks of training were treated with 4 applications, one per week, of 50.1 intra-articularly of the mixture of (v) matricins + polymerized collagen (5:5, vol:vol). Three animals were sacrificed in the first and seventh weeks post-treatment.

[0152] Early OA model. Fifteen rats underwent partial meniscectomy of the right rear knee, 3 weeks of training and treatment with 4 intra-articular applications, one per week of 50.1 of:

[0153] (i) placebo (vehicle: citrate buffer; n=3);

[0154] (ii) polymeric type I atelopeptide collagen (polymerized collagen) (n=3);

[0155] (iii) the mixture of matricins with polymeric type I atelopeptide collagen (matricins + polymerized collagen 1 : 9 , vol : vol ) ( n=3 );

[0156] (iv) 1 mixture of matrixes + polymerized collagen 3: 7 (vol: vol, n=3); and

[0157] (v) 1 a mixture of matricins + polymerized collagen 5 : 5 ( vol : vol , n=3 ) .

[0158] The animals were sacrificed in the first week post-treatment.

[0159] Long-term OA model. Fifteen rats underwent partial meniscectomy of the right rear knee, 3 weeks of training and treatment with 4 intra-articular applications, one per week of 50.1 of the treatments(iv). The animals were sacrificed in the seventh week post-treatment.

[0160] From all animals, the spleen, lymph nodes, joint tissue, liver, lungs, kidneys, and heart were obtained in order to assess inflammatory infiltrate and predominant cell populations, cytokine expression, and ECM turnover.

[0161] Clinical measurements were performed under baseline conditions and then weekly for 15 weeks. Leg thickness increase was measured with an electronic caliper. Weight and temperature were determined. Radiographic progression was assessed at weeks 1 and 7 post-treatment in a blinded manner by the same observer. Tissue architecture and proteoglycan content were determined using the O-Safranin staining technique (red-stained area). Immunofluorescence was used to determine the expression of MMP-13 (cartilage degradation), type I collagen (fibrocartilage formation), and type II collagen (hyaline cartilage formation) by quantifying the number of fluorescent pixels (fluorescence intensity).

[0162] Clinical and demographic characteristics.

[0163] With the exception of the placebo-treated OA group (i), all other groups (ii-v) showed a significant increase in weight (Figure 15, A). No differences were found between the study groups. Only the placebo-treated OA rats (i) showed an increase in temperature at week 8 when compared to the other study groups (ii-v) (Figure 15, C). The thickness of the right hind knee was significantly greater from week 11 onwards in the placebo-treated OA rats (i) compared to the other study groups (ii-v) (Figure 15, B).

[0164] Figure 15. Effect of the matrix mixture with polymeric atelopeptide type I collagen in the early and long-standing osteoarthritis (OA) model. Fifty units of each treatment were administered intra-articularly once a week for 4 weeks in the right hind leg (n = 6, per group) • (A) Weight, (B) knee thickness in early OA (9 a week) and long evolution (15 a week) and (C) temperature of the rats. Data represent mean ± standard error (SEM). * P<0.05.

[0165] Radiological and macroscopic findings of the effect of the mixture of matricins and polymeric type I atelopeptide collagen in the joint cavity.

[0166] Radiographic assessment was scored on a scale of 0 (no damage) to 3 (severe damage), according to the consensus of two blinded evaluators. The control group without meniscectomy and without training; the control group without meniscectomy, with surgery and with training; the toxicity group; and the short- and long-standing OA groups treated with (v) the matrix + 5:5 polymerized collagen mixture preserved the normal joint cavity structure (grade 0; in A of Fig. 16). Short- and long-standing OA rats treated with (ii) polymerized collagen had a grade of 0–1. The groups treated with the (iii) matrix + 1:9 polymerized collagen and (iv) matrix + 3:7 polymerized collagen mixtures had a severity score of 1–2. Whereas rats with OA treated with placebo (i) severe joint cavity damage with a score of 3.The OA was accompanied by increases in the volume, thickness, and contour of the cortical plate, alterations in the bone mineralization status, changes in the architecture of the subchondral trabecular bone and bone mass, the formation of bone cysts, the appearance of osteophytes, and narrowing of the joint space. Tilting with bone contact was also observed (in A of Fig. 16). The radiological findings correlated directly with the macroscopic findings, with the mixture of (v) matricins + 5:5 (vol:vol) polymerized collagen remaining the best treatment, followed by (ii) polymerized collagen (in B of Fig. 16).

[0167] Figure 16. The mixture of matricins with polymeric atelopeptide type I collagen (5:5) preserves the radiographic and macroscopic characteristics of a healthy joint cavity in the early and long-standing osteoarthritis (OA) model. (In A of Figure 16) Radiographs of the knees of the right hind legs that underwent partial meniscectomy and were treated with placebo, both in the short and long term, show loss of joint space (white arrow) and chondrocalcinosis; while treatment with the mixture of matricins + polymerized collagen (5:5) preserves the joint cavity. (In B of Figure 16) The cartilage surface of the control groups without OA shows condyles with a white, smooth, and shiny surface. The cartilage surface of the OA group treated with placebo appears opalescent, opaque, rough, and eroded. Cartilage loss also occurs in some regions (black circle).Joints treated with the mixture of matricins + polymerized collagen (5:5) (one application per week / 4 weeks, via intra-articular injection), exhibit an architecture similar to normal.

[0168] Histological findings of the effect of the mixture of matricins and polymeric type 1 ate lopeptide collagen on the proteoglycan content of hyaline cartilage.

[0169] The knee cartilage from the two control groups and the toxicity group showed the typical chondrocytes of the three main tissue zones: flattened chondrocytes in the superficial zone, rounded secretory chondrocytes in the middle zone, and hypertrophic chondrocytes in the deep zone. Homogeneous proteoglycan expression was observed, covering all three cartilage zones (Fig. 17).

[0170] The cartilage of rats with OA treated with placebo showed deep fibrillations, and the surface zone was lost in some regions of the cartilage. The number of fibroblasts and fibrous tissue increased. A decrease in the number of chondrocytes was also observed; hypertrophic chondrocytes appeared shrunken, and empty lacunae increased. Furthermore, a progressive and severe reduction in proteoglycan content was observed. The ECM showed a disorganized architecture, and the surface became fibrillar. The junctional zone appeared as a blurred region and in many cases was not visible (Fig. 17).

[0171] The early and long evolution OA groups treated with 50 pL of (v) the mixture of matricins + 5:5 polymerized collagen and (ii) polymerized collagen exhibited cartilage with histological characteristics and proteoglycan content similar to those observed in the control groups (Fig. 17).

[0172] It is important to mention that the contralateral leg of these rats did not show OA-related degeneration (data not shown).

[0173] Figure 17. Preservation of proteoglycan content in the mid-zone of cartilage in early and long-standing osteoarthritis, treated with the mixture of matricins and polymeric atelopeptide type I collagen 5:5. The arrows show the fibrillated surface of the tissue, the arrowhead points to a group of chondrocytes, and the star indicates the loss of proteoglycans (stained red). Magnification at X400.

[0174] Effect of the matrix mixture with polymeric atelopeptide type I collagen on type II collagen expression by chondrocytes. Chondrocytes in the cartilage of the control groups showed significantly higher type II collagen expression compared to the cartilage of rats with OA treated with (i) placebo (in A and B of Fig. 18). The early and long-standing OA group treated with 4 intra-articular administrations of 50 pL of (v) the matrix mixture + polymerized collagen in a 5:5 (vol:vol) ratio and (ii) polymerized collagen showed a similar number of type II collagen-producing chondrocytes to those observed in the control groups (In A and B of Fig. 18). However, it should be noted that the best result was obtained with (v) 1 a mixture of matricins + polymerized collagen 5 : 5 (In B of Fig. 18 ).

[0175] Figure 18. Effect of the matrix mixture with 5:5 polymerized atelopeptide type I collagen on the increase in type II collagen expression by cartilage chondrocytes. (A) Type II collagen was labeled with FITC (green) and nuclei were counterstained with DAPI (blue). (B) The graph shows a statistically significant decrease in type II collagen in the cartilage of OA rats treated with placebo versus the training control, surgical control, toxicity model, or early and long-term OA treatment group with 50 pL of the (v) matrix mixture + 5:5 polymerized collagen or of the (ii) polymerized collagen (*P<0.05). Data from three independent experiments (n=6). Results are expressed as mean ± standard deviation (SD).

[0176] Negative regulatory effect of the mixture of matricins and polymeric atelopeptide type I collagen on the expression of type 1 collagen by chondrocytes.

[0177] Chondrocytes in the cartilage of the control groups showed significantly lower expression of type I collagen compared to the cartilage of placebo-treated OA rats (fibrocartilage; In A and B of Fig.

[0178] 19)

[0179] The early and long-standing OA groups treated with 4 intra-articular administrations of 50 pL of (v) the matrix + polymerized collagen mixture in a 5:5 (vol:vol) ratio and (ii) polymerized collagen showed a similar number of type I collagen-producing chondrocytes to those observed in the control groups (In A and B of Fig. 19). However, it should be noted that the best result was obtained with (v) 1 matrix + polymerized collagen mixture 5:5 (vol:vol) (In B of Fig. 19).

[0180] Figure 19. Negative regulatory effect of the matrix mixture with polymeric atelopeptide type I collagen on fibrocartilage formation in rats with osteoarthritis (OA). (In A of Figure 19) Expression of type I collagen in rat articular cartilage. Type I collagen was labeled with FITC (green) and nuclei were counterstained with DAPI (blue). (In B of Figure 19) The graph shows a statistically significant increase in type I collagen in the cartilage of rats with OA treated with placebo versus exercise control, surgical control, toxicity model, or the early and long-term OA treatment group treated with 4 intra-articular administrations, once a week, with 50 pL of the matrix + polymerized collagen mixture or polymerized collagen alone (*P<0.05). Data from three independent experiments (n = 6). Results are expressed as mean ± standard deviation (SD).

[0181] Effect of the mixture of matricins and polymeric atelopeptide type I collagen on the expression of MMP-13 by chondrocytes.

[0182] Chondrocytes from the control groups showed significantly lower expression of MMP-13 compared to cartilage from placebo-treated OA rats, which is associated with less destruction of articular cartilage (in A and B of Fig. 20).

[0183] The early and long-standing OA groups treated with 4 intra-articular administrations of 50 pL of (v) the mixture of matricins + polymerized collagen in a 5:5 (vol:vol) ratio and (ii) polymerized collagen showed a similar number of MMP-13 producing chondrocytes to those observed in the control groups (in A and B of Fig. 20).

[0184] Adverse effects.

[0185] There was no evidence of adverse events following the placebo injection or the different treatments throughout the study. The assessment of adverse events included the determination of pseudoseptic reaction, crystal arthritis, swelling, local skin reactions, etc. The only adverse event observed was pain lasting less than 15 minutes at the injection site.

[0186] Figure 20. Mixture of matricins with polymeric atelopeptide type I collagen decreases the expression of metalloproteinase type 13 (MMP-13) by chondrocytes. (In A of Figure 20) MMP-13 expression in rat articular cartilage. MMP-13 was labeled with FITC (green) and nuclei were counterstained with DAPI (blue). (In B of Figure 20) The graph shows a significant increase in MMP-13 in the cartilage of OA rats treated with placebo versus the training control, surgical control, toxicity model, or the early and long-term OA treatment group with 4 intra-articular administrations, once a week, with 50 pL of the (v) matrix + polymerized collagen mixture or the (ii) polymerized collagen (*P<0.05). Data from three independent experiments (n = 6). Results are expressed as mean ± standard deviation (SD).

[0187] In conclusion, 4 intra-articular administrations of the mixture of matricins + polymerized collagen in a 5:5 (vol:vol) ratio is an effective treatment that modifies the progression of OA, inducing chondroprotection and high-quality cartilage (hyaline cartilage), whose function is not only restricted to providing protection, lubrication and mechanical stability to the collagen network, cells and articular surfaces, but also positively regulates the synthesis of highly sulfated proteoglycans and type II collagen and negatively regulates the expression of type I collagen, as well as the collagenolytic activity mediated by MMP13. The mixture of matricins + polymerized collagen in a 5:5 (vol:vol) ratio was safe as determined in the toxicity model, well tolerated and without adverse events, except for pain lasting less than 15 min at the injection site.

[0188] The invention has been described based on a preferred embodiment; however, it is evident that the invention accepts numerous changes and modifications, all within the spirit and scope of the invention. The present invention therefore includes all such changes and modifications, being limited only by the terms of the claims.

Claims

RE VIND CAC IONE S 1. A chemical composition based on a mixture of matrixes and polymeric type I atelopeptide collagen.

2. A composition according to claim 1, wherein the matrices are selected from collagens and elastocins.

3. A chemical composition according to claim 1, wherein the collagens have 0.15-0.2% protein nitrogen.

4. A chemical composition according to claim 1, wherein the astocins have 1.5-2.0% protein nitrogen.

5. A chemical composition according to claim 1, comprising a mixture of matricins and polymeric type I atelopeptide collagen in a volume-to-volume ratio from 0.1-9.9 to 9.9-0.1 of each component, respectively.

6. A chemical composition according to claim 1, comprising a mixture of matricins, specifically collagens and elastocins.

7. Process for preparing a composition based on matricins obtained by hydrolysis by heat at 105-125°C for 30 minutes, at a pressure between 13 and 25 pounds, at a slightly acidic pH of a mixture of water-soluble bovine elastin and native porcine collagen type I (3:1 ratio) to obtain collagen hydrolysates (collagenocins) that vary in molecular weight from 1000 to 25,000 Da, while those of elastin (elastoins) range from 200 to 150,000; with a porcine atelopeptide type I collagen in a buffer solution, and a vinyl polymer with acidic pH, this polymer being selected from a group consisting of polyvinyl alcohol, polyvinyl chloride, polyvinylpyrrolidone or any other pharmacologically acceptable vinyl polymer; this collagen is mixed with the vinyl polymer and the resulting composition is sterilized, and crosslinked by gamma radiation.

8. A process according to claim 7, wherein the collagen is "native" type I collagen (5 to 30 mg), is solubilized in a buffer solution and polymer, the mixture is irradiated with gamma rays obtaining a controlled crosslinking between the collagen and the vinyl polymer.

9. A process according to claim 7 or 8, wherein the polymer is selected from the group consisting of polyvinyl alcohol, polyvinyl chloride, polyvinylpyrrolidone or a pharmacologically acceptable vinyl polymer.

10. A process according to any of claims 7 to 9, wherein the polymer is in a proportion of between 5 and 50% of the composition.

11. Use of a composition of collagens, elastocins and polymeric type I atelopeptide collagen, wherein the composition is applicable in the pharmaceutical therapeutic or cosmetic areas.

12. Use in accordance with claim 11, wherein the composition is an inflammation modulator, inducer of peripheral immune tolerance, antifibrotic, fibrolytic and inducer of tissue regeneration.

13. Use of a composition of collagenocins, elastocins and polymeric type I atelopeptide collagen, according to any of claims 1 to 6, for preparing a medicament in the treatment of osteoarthritis (OA), rotator cuff tendinosis, epicondylitis, bursitis, tenosynovitis, tendinitis, degenerative tendinosis, partial tendon rupture, rheumatoid arthritis (RA), scleroderma (SSc), scarring, keloid or hypertrophic scarring, pulmonary fibrosis, chronic obstructive pulmonary emphysema, hepatic cirrhosis, valvular heart disease, diabetic ulcers, pressure ulcers, tendon sclerosis, fracture healing, achalasia and other similar conditions, which have as a common denominator the presence of acute inflammation, chronic inflammation, hyperinflammation, cytokine release syndrome and fibrosis.

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