WO2020094902A1 - Inhibitors of tgf-β1 and endoglin producers for use in the treatment of epidermolysis bullosa - Google Patents
Inhibitors of tgf-β1 and endoglin producers for use in the treatment of epidermolysis bullosa Download PDFInfo
- Publication number
- WO2020094902A1 WO2020094902A1 PCT/ES2019/070757 ES2019070757W WO2020094902A1 WO 2020094902 A1 WO2020094902 A1 WO 2020094902A1 ES 2019070757 W ES2019070757 W ES 2019070757W WO 2020094902 A1 WO2020094902 A1 WO 2020094902A1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- tgf
- raloxifene
- endoglin
- treatment
- levels
- Prior art date
Links
- 238000011282 treatment Methods 0.000 title claims abstract description 20
- 108010036395 Endoglin Proteins 0.000 title claims abstract description 18
- 102000012085 Endoglin Human genes 0.000 title claims abstract description 18
- 239000003112 inhibitor Substances 0.000 title claims description 6
- 206010014989 Epidermolysis bullosa Diseases 0.000 title abstract description 23
- 230000037361 pathway Effects 0.000 claims abstract description 8
- 239000008194 pharmaceutical composition Substances 0.000 claims abstract description 5
- GZUITABIAKMVPG-UHFFFAOYSA-N raloxifene Chemical compound C1=CC(O)=CC=C1C1=C(C(=O)C=2C=CC(OCCN3CCCCC3)=CC=2)C2=CC=C(O)C=C2S1 GZUITABIAKMVPG-UHFFFAOYSA-N 0.000 claims description 26
- 229960004622 raloxifene Drugs 0.000 claims description 26
- 150000001875 compounds Chemical class 0.000 claims description 20
- 229960004308 acetylcysteine Drugs 0.000 claims description 8
- UCJGJABZCDBEDK-UHFFFAOYSA-N bazedoxifene Chemical compound C=1C=C(OCCN2CCCCCC2)C=CC=1CN1C2=CC=C(O)C=C2C(C)=C1C1=CC=C(O)C=C1 UCJGJABZCDBEDK-UHFFFAOYSA-N 0.000 claims description 8
- 229960000817 bazedoxifene Drugs 0.000 claims description 8
- PWKSKIMOESPYIA-BYPYZUCNSA-N L-N-acetyl-Cysteine Chemical compound CC(=O)N[C@@H](CS)C(O)=O PWKSKIMOESPYIA-BYPYZUCNSA-N 0.000 claims description 7
- 208000017520 skin disease Diseases 0.000 claims description 6
- 206010053177 Epidermolysis Diseases 0.000 claims description 5
- 208000034607 Kindler epidermolysis bullosa Diseases 0.000 claims description 2
- 201000004290 Kindler syndrome Diseases 0.000 claims description 2
- 239000000546 pharmaceutical excipient Substances 0.000 claims description 2
- 239000000203 mixture Substances 0.000 claims 2
- 229940079593 drug Drugs 0.000 abstract description 9
- 239000003814 drug Substances 0.000 abstract description 9
- 102000046299 Transforming Growth Factor beta1 Human genes 0.000 abstract description 2
- 101800002279 Transforming growth factor beta-1 Proteins 0.000 abstract description 2
- 230000002401 inhibitory effect Effects 0.000 abstract description 2
- 230000004936 stimulating effect Effects 0.000 abstract 1
- 210000002950 fibroblast Anatomy 0.000 description 21
- 108090000623 proteins and genes Proteins 0.000 description 14
- 230000014509 gene expression Effects 0.000 description 12
- 102000004169 proteins and genes Human genes 0.000 description 11
- 206010016654 Fibrosis Diseases 0.000 description 10
- 230000004761 fibrosis Effects 0.000 description 10
- 108010035532 Collagen Proteins 0.000 description 9
- 102000008186 Collagen Human genes 0.000 description 9
- 229920001436 collagen Polymers 0.000 description 9
- 108010037362 Extracellular Matrix Proteins Proteins 0.000 description 8
- 239000002083 C09CA01 - Losartan Substances 0.000 description 7
- 102000010834 Extracellular Matrix Proteins Human genes 0.000 description 7
- KJJZZJSZUJXYEA-UHFFFAOYSA-N losartan Chemical compound CCCCC1=NC(Cl)=C(CO)N1CC1=CC=C(C=2C(=CC=CC=2)C=2[N]N=NN=2)C=C1 KJJZZJSZUJXYEA-UHFFFAOYSA-N 0.000 description 7
- 229960004773 losartan Drugs 0.000 description 7
- 102100025751 Mothers against decapentaplegic homolog 2 Human genes 0.000 description 6
- 101710143123 Mothers against decapentaplegic homolog 2 Proteins 0.000 description 6
- 102000007000 Tenascin Human genes 0.000 description 6
- 108010008125 Tenascin Proteins 0.000 description 6
- 230000015572 biosynthetic process Effects 0.000 description 6
- 230000008602 contraction Effects 0.000 description 6
- 230000007423 decrease Effects 0.000 description 6
- 201000010099 disease Diseases 0.000 description 6
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 6
- 102000005962 receptors Human genes 0.000 description 6
- 108020003175 receptors Proteins 0.000 description 6
- 238000004519 manufacturing process Methods 0.000 description 5
- 238000000034 method Methods 0.000 description 5
- 238000003786 synthesis reaction Methods 0.000 description 5
- 238000012360 testing method Methods 0.000 description 5
- 108060003393 Granulin Proteins 0.000 description 4
- 102100037765 Periostin Human genes 0.000 description 4
- 101710199268 Periostin Proteins 0.000 description 4
- 230000004913 activation Effects 0.000 description 4
- 238000003556 assay Methods 0.000 description 4
- 210000004027 cell Anatomy 0.000 description 4
- 230000001413 cellular effect Effects 0.000 description 4
- 230000000694 effects Effects 0.000 description 4
- 210000002744 extracellular matrix Anatomy 0.000 description 4
- 230000002206 pro-fibrotic effect Effects 0.000 description 4
- 230000008569 process Effects 0.000 description 4
- 210000002966 serum Anatomy 0.000 description 4
- 238000002965 ELISA Methods 0.000 description 3
- 108010067306 Fibronectins Proteins 0.000 description 3
- 102000016359 Fibronectins Human genes 0.000 description 3
- 206010061218 Inflammation Diseases 0.000 description 3
- 206010028980 Neoplasm Diseases 0.000 description 3
- -1 TGFi Proteins 0.000 description 3
- 239000000512 collagen gel Substances 0.000 description 3
- 238000011161 development Methods 0.000 description 3
- 238000003745 diagnosis Methods 0.000 description 3
- 239000000835 fiber Substances 0.000 description 3
- 230000004054 inflammatory process Effects 0.000 description 3
- 239000003446 ligand Substances 0.000 description 3
- 239000011159 matrix material Substances 0.000 description 3
- 230000007246 mechanism Effects 0.000 description 3
- 230000035772 mutation Effects 0.000 description 3
- 230000002018 overexpression Effects 0.000 description 3
- 230000026731 phosphorylation Effects 0.000 description 3
- 238000006366 phosphorylation reaction Methods 0.000 description 3
- 230000009467 reduction Effects 0.000 description 3
- 230000019491 signal transduction Effects 0.000 description 3
- 210000003491 skin Anatomy 0.000 description 3
- 230000001225 therapeutic effect Effects 0.000 description 3
- 102000012422 Collagen Type I Human genes 0.000 description 2
- 108010022452 Collagen Type I Proteins 0.000 description 2
- 108700037244 Complement Component 7 Deficiency Proteins 0.000 description 2
- 208000031953 Hereditary hemorrhagic telangiectasia Diseases 0.000 description 2
- 108090001005 Interleukin-6 Proteins 0.000 description 2
- 102000009618 Transforming Growth Factors Human genes 0.000 description 2
- 239000000090 biomarker Substances 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- DDRJAANPRJIHGJ-UHFFFAOYSA-N creatinine Chemical compound CN1CC(=O)NC1=N DDRJAANPRJIHGJ-UHFFFAOYSA-N 0.000 description 2
- 230000001404 mediated effect Effects 0.000 description 2
- 108020004999 messenger RNA Proteins 0.000 description 2
- 230000008506 pathogenesis Effects 0.000 description 2
- 230000000144 pharmacologic effect Effects 0.000 description 2
- 238000002560 therapeutic procedure Methods 0.000 description 2
- 230000002792 vascular Effects 0.000 description 2
- MZOFCQQQCNRIBI-VMXHOPILSA-N (3s)-4-[[(2s)-1-[[(2s)-1-[[(1s)-1-carboxy-2-hydroxyethyl]amino]-4-methyl-1-oxopentan-2-yl]amino]-5-(diaminomethylideneamino)-1-oxopentan-2-yl]amino]-3-[[2-[[(2s)-2,6-diaminohexanoyl]amino]acetyl]amino]-4-oxobutanoic acid Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H](CC(O)=O)NC(=O)CNC(=O)[C@@H](N)CCCCN MZOFCQQQCNRIBI-VMXHOPILSA-N 0.000 description 1
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 1
- 101150084750 1 gene Proteins 0.000 description 1
- 101150056204 COL7A1 gene Proteins 0.000 description 1
- 208000005623 Carcinogenesis Diseases 0.000 description 1
- 201000009030 Carcinoma Diseases 0.000 description 1
- 108010017377 Collagen Type VII Proteins 0.000 description 1
- 102000004510 Collagen Type VII Human genes 0.000 description 1
- 108090000695 Cytokines Proteins 0.000 description 1
- 102000004127 Cytokines Human genes 0.000 description 1
- 108090000738 Decorin Proteins 0.000 description 1
- 102000004237 Decorin Human genes 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 208000032843 Hemorrhage Diseases 0.000 description 1
- 101100496573 Homo sapiens COL7A1 gene Proteins 0.000 description 1
- 208000002682 Hyperkalemia Diseases 0.000 description 1
- 208000013016 Hypoglycemia Diseases 0.000 description 1
- 208000001953 Hypotension Diseases 0.000 description 1
- 108010052285 Membrane Proteins Proteins 0.000 description 1
- 102100025744 Mothers against decapentaplegic homolog 1 Human genes 0.000 description 1
- 101100490437 Mus musculus Acvrl1 gene Proteins 0.000 description 1
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 description 1
- 102000001253 Protein Kinase Human genes 0.000 description 1
- 238000011529 RT qPCR Methods 0.000 description 1
- 208000035977 Rare disease Diseases 0.000 description 1
- 208000037014 Rare skin disease Diseases 0.000 description 1
- 208000001647 Renal Insufficiency Diseases 0.000 description 1
- 101700032040 SMAD1 Proteins 0.000 description 1
- 108060008682 Tumor Necrosis Factor Proteins 0.000 description 1
- 102000000852 Tumor Necrosis Factor-alpha Human genes 0.000 description 1
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 description 1
- 208000012886 Vertigo Diseases 0.000 description 1
- QQPGGBNMTNDKEY-UHFFFAOYSA-N [2-butyl-5-chloro-3-[[4-[2-(2-trityltetrazol-5-yl)phenyl]phenyl]methyl]imidazol-4-yl]methanol Chemical compound CCCCC1=NC(Cl)=C(CO)N1CC1=CC=C(C=2C(=CC=CC=2)C2=NN(N=N2)C(C=2C=CC=CC=2)(C=2C=CC=CC=2)C=2C=CC=CC=2)C=C1 QQPGGBNMTNDKEY-UHFFFAOYSA-N 0.000 description 1
- 230000001594 aberrant effect Effects 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 239000012190 activator Substances 0.000 description 1
- 239000000654 additive Substances 0.000 description 1
- 239000000853 adhesive Substances 0.000 description 1
- 230000001070 adhesive effect Effects 0.000 description 1
- 238000004873 anchoring Methods 0.000 description 1
- 208000007502 anemia Diseases 0.000 description 1
- 239000003963 antioxidant agent Substances 0.000 description 1
- 230000003078 antioxidant effect Effects 0.000 description 1
- 206010003549 asthenia Diseases 0.000 description 1
- 210000002469 basement membrane Anatomy 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 230000033228 biological regulation Effects 0.000 description 1
- 230000000740 bleeding effect Effects 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 201000011510 cancer Diseases 0.000 description 1
- 230000036952 cancer formation Effects 0.000 description 1
- 239000004202 carbamide Substances 0.000 description 1
- 231100000504 carcinogenesis Toxicity 0.000 description 1
- 230000003915 cell function Effects 0.000 description 1
- 229940109239 creatinine Drugs 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 230000002950 deficient Effects 0.000 description 1
- 210000004207 dermis Anatomy 0.000 description 1
- 208000002173 dizziness Diseases 0.000 description 1
- 230000004064 dysfunction Effects 0.000 description 1
- 210000002889 endothelial cell Anatomy 0.000 description 1
- 230000003511 endothelial effect Effects 0.000 description 1
- 229940011871 estrogen Drugs 0.000 description 1
- 239000000262 estrogen Substances 0.000 description 1
- 230000001076 estrogenic effect Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 239000000499 gel Substances 0.000 description 1
- 230000004547 gene signature Effects 0.000 description 1
- 230000002218 hypoglycaemic effect Effects 0.000 description 1
- 230000036543 hypotension Effects 0.000 description 1
- 238000003018 immunoassay Methods 0.000 description 1
- 230000006698 induction Effects 0.000 description 1
- 230000002757 inflammatory effect Effects 0.000 description 1
- 208000014674 injury Diseases 0.000 description 1
- 108010028309 kalinin Proteins 0.000 description 1
- 201000006370 kidney failure Diseases 0.000 description 1
- 239000003550 marker Substances 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 230000009456 molecular mechanism Effects 0.000 description 1
- 238000012544 monitoring process Methods 0.000 description 1
- 238000010172 mouse model Methods 0.000 description 1
- 208000015122 neurodegenerative disease Diseases 0.000 description 1
- 238000010606 normalization Methods 0.000 description 1
- 229940000673 orphan drug Drugs 0.000 description 1
- 239000002859 orphan drug Substances 0.000 description 1
- 230000001575 pathological effect Effects 0.000 description 1
- 230000007170 pathology Effects 0.000 description 1
- 230000035778 pathophysiological process Effects 0.000 description 1
- 230000007310 pathophysiology Effects 0.000 description 1
- 238000011458 pharmacological treatment Methods 0.000 description 1
- 230000030716 positive regulation of phosphorylation Effects 0.000 description 1
- 229910052700 potassium Inorganic materials 0.000 description 1
- 239000011591 potassium Substances 0.000 description 1
- 238000004393 prognosis Methods 0.000 description 1
- 230000035755 proliferation Effects 0.000 description 1
- 108060006633 protein kinase Proteins 0.000 description 1
- 238000003753 real-time PCR Methods 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 208000011728 renal alteration Diseases 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 238000003757 reverse transcription PCR Methods 0.000 description 1
- 108020004418 ribosomal RNA Proteins 0.000 description 1
- 239000000523 sample Substances 0.000 description 1
- 230000002269 spontaneous effect Effects 0.000 description 1
- 230000006641 stabilisation Effects 0.000 description 1
- 238000011105 stabilization Methods 0.000 description 1
- 208000011580 syndromic disease Diseases 0.000 description 1
- 230000002103 transcriptional effect Effects 0.000 description 1
- 229940099456 transforming growth factor beta 1 Drugs 0.000 description 1
- 230000008733 trauma Effects 0.000 description 1
- 210000005167 vascular cell Anatomy 0.000 description 1
- 239000013598 vector Substances 0.000 description 1
- 231100000889 vertigo Toxicity 0.000 description 1
- 102000009310 vitamin D receptors Human genes 0.000 description 1
- 108050000156 vitamin D receptors Proteins 0.000 description 1
Classifications
-
- 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
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
- A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
- A61K31/445—Non condensed piperidines, e.g. piperocaine
- A61K31/4523—Non condensed piperidines, e.g. piperocaine containing further heterocyclic ring systems
- A61K31/4535—Non condensed piperidines, e.g. piperocaine containing further heterocyclic ring systems containing a heterocyclic ring having sulfur as a ring hetero atom, e.g. pizotifen
-
- 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
- A61K31/185—Acids; Anhydrides, halides or salts thereof, e.g. sulfur acids, imidic, hydrazonic or hydroximic acids
- A61K31/19—Carboxylic acids, e.g. valproic acid
- A61K31/195—Carboxylic acids, e.g. valproic acid having an amino group
- A61K31/197—Carboxylic acids, e.g. valproic acid having an amino group the amino and the carboxyl groups being attached to the same acyclic carbon chain, e.g. gamma-aminobutyric acid [GABA], beta-alanine, epsilon-aminocaproic acid or pantothenic acid
- A61K31/198—Alpha-amino acids, e.g. alanine or edetic acid [EDTA]
-
- 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
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/55—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having seven-membered rings, e.g. azelastine, pentylenetetrazole
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P17/00—Drugs for dermatological disorders
Definitions
- TGF-B1 inhibitors and endoqlin producers for use in the treatment of hulose
- the present invention relates to the repositioning of TGF-bI pathway modulating drugs and stimulators of endoglin expression, particularly raioxifene and apeledoxifene, for use in the treatment of fibrosis such as epidermolysis hullosa (EB).
- EB epidermolysis hullosa
- TGF-bI transforming growth factor-beta 1
- Epidermoiisis bullosa comprises a clinically and genetically heterogeneous group of rare skin diseases characterized by the formation of skin blisters, spontaneous or induced by minimal trauma (Fine el al 2009: Fine, Jo-David, Hintner, Helmut. Life with Epidermolysis Bullosa (EB): Etiology, Diagnosis, Multidiscipiinary Care and Therapy. SpringerWienNewYork. 2009, 338 p ISBN: 978-3-211-792-70-4).
- EBS simple
- EBJ joint!
- EPD dystrophic
- EBM mixed
- EBDR recessive EBD form
- EBDD dominant EBD form
- C7 Collagen Vil
- This protein is the main component of the anchoring fibrils that link type I collagen of the dermis with subepidermal basement membrane proteins such as laminin 332 acting as a true dermo-epidermal adhesive not only on the skin but also on mucosae such as the esophageal.
- laminin 332 acting as a true dermo-epidermal adhesive not only on the skin but also on mucosae such as the esophageal.
- the absence or pronounced decrease of C7 is the main characteristic of EBDR and its determination establishes a primary molecular diagnosis of the pathology.
- TGF-bI receptor complex consists of the type II receptor (TpRIl), the type I receptor (ALK5) and the co-receptor Endoglin.
- Endoglin modulates the activation of phosphorylation of ALK5 by the T Ril receptor, after binding of the TGF-bI ligand (Santiba ⁇ ez et al 2011: Santiba ⁇ ez JF, Quintanilia M, Bernabeu G. TQR-b / TQR-b receptor system and its role in physiological and pathological conditions. ClinSci (Lond). 2011 Sep; 121 (6): 233-51).
- the excess production of the TGF-bI ligand can be counteracted by an increase in endoglin, so that the phosphorylation of the Smad2 / 3 coactivators and their target genes involved in fibrosis (collagen synthesis) is prevented.
- fibronectin extracellular matrix proteins.
- Nystróm (Nystróm et al. 2015: Nystróm A, Thriene K, Mittapalli V, Kern JS, KiristiD, Dengjel J & Brucker-Tuderman L. Losar ⁇ an ameliorates dystrophic epidermo ⁇ ysis hullosa and uncovers new disease mechanisms.
- EBDR disease model mice have an excess production of serum TGF-bI, and that treatment with losartan decreases its amount although the side effects of Losar ⁇ an may limit its therapeutic use (for example: anemia ; dizziness, vertigo; hypotension; renal alteration, renal failure; asthenia, fatigue; hyperkalemia, increased blood urea, serum creatinine and potassium; hypoglycemia).
- N-acetiicysteine Another compound that activates endoglin production and inhibits TGF-bI production is N-acetiicysteine (NAC).
- the antioxidant N-acetylcysteine (NAC) has a recognized therapeutic value in inflammation, fibrosis, endoteiial dysfunction and promotes the synthesis of a large number of genes by modulating signaling pathways of different kinases, a useful concept for the treatment of certain degenerative diseases (Zafaruliah M, Li WQ, Sylvester J, Ahmad M. Molecular mechanisms of N-acetylcysteine actions.Cell Mol Life Sel. 2003 Jan; 60 (1): 6 ⁇ 20)
- drugs that are effective and selective for the treatment of fibrosis and in particular of a rare disease such as epidermolysis hullosa (EB) that represent an increase in the quality of life of patients with these diseases.
- EB epidermolysis hullosa
- the present invention relates to a compound characterized in that it is an inhibitor of the TGF-bI pathway and an endoglin stimulator for use in the treatment of genodermatosis.
- the invention relates to the compound according to the previously defined use selected for raloxifene and apeledoxifene.
- the invention relates to the compound according to the use defined above, where the disease is selected from epidermolysis hullosa (EB).
- EB epidermolysis hullosa
- the invention relates to the compound according to the use defined above, where EB is selected from simple EB (EBS), junctional EB (EBJ), dystrophic EB (EBD) and Kindler's syndrome
- EBD is selected from recessive EBD (EBDR) and dominant EBD (EBDD).
- the invention relates to the compound according to the use defined above, where the compound is selected from raloxifene, apeledoxifene and N-acetiicysteine, preferably where the compound raloxifene and N-acetiicysteine, for the treatment of genodermatosis, preferably for the treatment of epidermolysis hullosa (EB), more preferably for the treatment of simple EB (BBS), junctional EB (EBJ), dystrophic EB (EBD) or kindier syndrome and even more preferably for the treatment of recessive EBD (EBDR) or dominant EBD ( EBDD).
- EB epidermolysis hullosa
- BVS simple EB
- EBJ junctional EB
- ESD dystrophic EB
- EBDR recessive EBD
- EBDD dominant EBD
- Another aspect of the present invention relates to a pharmaceutical composition
- a pharmaceutical composition comprising one or more compounds characterized in that they are inhibitors of the TGF-bI pathway and endogiine stimulators for use in the treatment of genodermatosis and one or more pharmaceutically acceptable excipients. for its previously defined use.
- the invention relates to the pharmaceutical composition defined above comprising raloxifene and N-acetylcysteine.
- the invention relates to the pharmaceutical composition defined above comprising apeledoxifene and N-acetylcysteine.
- the present invention demonstrates that the treatment of fibroblasts from EBDR patients with Raloxifene and NAC causes the overexpression of endogiin and could be associated with the observed repression of the levels of TGF-bI and its targets of cellular matrix.
- the results show that the levels of the active form of TGF-bI in the serum of the EB patients tested is a biomarker of the disease.
- the extracellular matrix genes it regulates type 1 collagen, tenascin, decorin, fibronectin, periostin, TGF-bI are repressed and the extracellular matrix would normalize.
- FIG. 1 Determination of the levels of active TGF-bI in 23 sera from patients with Dominant Recessive Epidermolysis Bullosa (EBDR). Active (A) and total (B) TGF-bI levels in healthy subjects and EBDR patients.
- EBDR Dominant Recessive Epidermolysis Bullosa
- FIG. 2 Raloxifene and NAC modulate levels of endogfin protein and IGF-b1.
- the effect of raloxifene (0.2hM) and NAC (HQQuIVI ⁇ on endoglin levels and total and active TGF-bI was analyzed by ELISA under serum deprivation conditions and in the presence of the drug, during 48h, by ELISA.
- Figures 2A, 2C and 2E represent the rate of change (fold change) in the levels of endoglin and TGF in the fibroblasts of 3 patients.
- Figures 2B, 2D and 2F represent the mean of these changes in said 3 patients, * p ⁇ 0, Q5; ** p ⁇ 0.001; *** p ⁇ 0.001.
- FIG. 3 Raloxifene and NAC regulate mRNA expression of proteins associated with fibrosis.
- A Endoglin and TGF-b RNA levels were measured in fibroblasts from patients cultured in the presence or absence of raloxifene (0.2nM) or NAC (100uM) for 48h
- B Expression levels of proteins associated with fibrosis (Tenascin C, Periostin, TGFi, TSP and! L-6) measured by real quantitative PCR (qRT-PCR) under the above conditions. Transcript levels were normalized with the ACTB ribosomal RNA control, * p ⁇ Q, 05; ** p ⁇ 0.001; *** p ⁇ 0.001.
- FIG. 4 Test of contractile capacity of fibroblasts from 1 EBDR patient in geys of collagen in the absence and presence of raloxifene and iosartan.
- FSG 5 Test of contractile capacity of fibroblasts from 4 patients of the EBDR cohort in collagen geies in the absence and presence of NAC.
- FIG. 8 Mechanisms of drugs under study in fibroblasts from EBDR patients, on the TGF-bI signaling pathway, by analyzing the expression of phosphorylated Smad2 and Smad 1/5 proteins.
- Human fibroblasts from patients with EBDR were transfected with the CAGA-luc (A and B) and BRE-luc (C and D) promoter-report vectors. Iuciferase activity was measured in untreated or raloxifene or NAC-treated cells for 48b and in the presence of TGF-b for the last 24b (E) or 3h (F). Representative WBs are shown.
- Figures A and C represent individual results from 3 patients, and B and D the fold induction ratio of themselves.
- Iuciferase assays were performed as indicated in Material and Methods. Smad2 or total Smad1 / 5 was used as loading control, b-actin also shows a similar result. The differences are statistically significant according to the Student test * P ⁇ 0.05; ** P ⁇ 0.01; *** P ⁇ 0.005.
- Example 1 Determination of ios levels of active TGF-bI in 23 sera from patients with Epiderolysis buliosa.
- TGF-bI concentration was determined by enzyme immunoassay (ELISA; R&D Systems). TGF-bI levels were determined in both non-acidified (TGF-bI -total) and acidified (TGF-b ⁇ -active) sera.
- Figure 1A shows the elevation of the levels of active TGF-bI in the sera of EBDR patients, compared to the practically absence (detectability) of the active protein in healthy controls.
- Figure 1 B shows ios total TGF-bI levels in the sera of the samples analyzed.
- TGF-bI TGF-bI
- Example 2 Pharmacological effect of raloxifene, N-acetylclstefna (NAC) and losartan in fibroblasts from EBDR patients.
- NAC N-acetylclstefna
- Figure 2 shows the detected levels of Endoglin proteins and active TGF-bI and total TGF-bI, respectively, in the fibroblasts of three EBDR patients treated with raloxifene, N-acetylcysteine (NAC) and losartan. While the total TGF-bI levels show different responses to the treatments according to the patient, Figure 2B shows the significant reduction in the levels of active TGF-bI with the three pharmacological treatments. This decrease in the levels of the active cytokine is especially conspicuous with the treatment with raloxifene in the fibroblasts of patients 17 and 31 and with CAP in patient 8. Losartan appears especially active in patient 31, who seems to be the most sensitive to any of the treatments carried out.
- Losartan has not been reported to increase endoglin expression levels or that raloxifene and NAC modulate levels of active TGF-bI in human fibroblasts.
- Example 3 Raloxifene, NAC, and losartan suppress transcriptional levels of matrix proteins Tenascin C, Periostin, and TGFi in fibroblasts from EBDR patients,
- EBDR pathogenesis postulates a connection between C7 deficiency and exacerbated bioavailability and overexpression of TGF-bI and genes regulated by this factor, including expression of cellular matrix proteins.
- Proteomic analyzes of samples from patients with EBDR reveal a high expression in the levels of several proteins related to inflammation mediated by TGF-bI and of proteins of the extracellular matrix such as Tenascin C (Guerra et al 2017).
- Figure 3 shows the decrease in mRNA levels by RT-PCR determinations of the cellular matrix proteins Tenascin C, Periostin, TGFi and Thromboespondin-1 (TSP1) as well as that of the inflammatory marker cytokine IL-6.
- Example 4 Raloxifene inhibits fibroblast proliferation surface contraction of an EBDR patient in collagen gels.
- the collagen contraction test (Ferrer-Mayorga et ai 2017: Ferrer- Mayorga G, et a !. Vitamin D receptor expression and associated gene signature in tumor stromal fibrobiasts predict clinicai outcome in colorectai cancer, Gut 2017, 66: 1449-1462 ) is used to explore the activation of fibroblasts and consequent reduction of the surface occupied by cells in culture, in the absence of adhesion to the supports of the walls of the assay plate.
- Raloxifene inhibits one of the profibrotic properties, such as the contraction of collagen gel fibers (see Figure 4), which is an illustrative characteristic of the activation of fibroblasts.
- Example 5 ISI-acetylcysteine inhibits the collagen fibers contraction of fibroblasts in collagen gels from four EBDR patients.
- the collagen contraction assay (Ferrer-Mayorga et al 2017) is used to explore the activation of ios fibroblasts by reducing the surface occupied by cells in culture, in the absence of adhesion to the walls of the assay plate (see control panel in Figure 5).
- N-acetylcysteine inhibits one of the profibrotic properties such as the contraction of the collagen fibers in gel, which is an illustrative characteristic of the activation of the fibroblasts ( Figure 5).
- Example 6 Raloxifene and MAC inhibit phosphorylation levels of Smad2 / 3 and Smad 1/5 phosphorylated.
- Raloxifene and NAC appear to affect the TGF-b / Smad signaling pathway differently than Losar ⁇ an, according to the results of Nystrom et al.): While raloxifene appears to selectively inhibit the Smad2 / 3 pathway, ia NAC inhibits Both routes indiscriminately, Smad 2/3 and Smad 1/5.
Landscapes
- Health & Medical Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Pharmacology & Pharmacy (AREA)
- Veterinary Medicine (AREA)
- Medicinal Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Public Health (AREA)
- Epidemiology (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Engineering & Computer Science (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Dermatology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Organic Chemistry (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention relates to a drug for inhibiting the TGF-β1 pathway and stimulating endoglin, or pharmaceutical composition thereof, for use in the treatment of genodermatoses, such as epidermolysis bullosa (EB).
Description
Inhibidores de TGF-B1 y productores de endoqlina para su uso en el tratamiento de hullosaTGF-B1 inhibitors and endoqlin producers for use in the treatment of hulose
La presente invención se refiere ai reposicionamiento de fármacos moduladores de ¡a ruta de TGF-bI y estimuladores de la expresión de endoglina, particularmente raioxifeno y bazedoxifeno, para su uso en el tratamiento de fibrosis tales como la epidermóiisis hullosa (EB). The present invention relates to the repositioning of TGF-bI pathway modulating drugs and stimulators of endoglin expression, particularly raioxifene and bazedoxifene, for use in the treatment of fibrosis such as epidermolysis hullosa (EB).
ANTECEDENTES DE LA INVENCIÓN BACKGROUND OF THE INVENTION
Las proteínas que integran el factor de crecimiento transformante-beta 1 (TGF-bI) regulan la función celular y tienen un papel clave en la carcinogénesis y la fisiopatología de las fibrosis. The proteins that make up transforming growth factor-beta 1 (TGF-bI) regulate cell function and play a key role in the carcinogenesis and pathophysiology of fibrosis.
La Epidermóiisis ampollosa (EB; OMIM: 120120) comprende a un grupo clínica y genéticamente heterogéneo de enfermedades raras de piel caracterizadas por la formación de ampollas cutáneas, espontáneas o inducidas por un trauma mínimo (Fine el al 2009: Fine, Jo-David, Hintner, Helmut. Life with Epidermolysis Bullosa (EB): Etiology, Diagnosis, Multidiscipiinary Care and Therapy. SpringerWienNewYork. 2009, 338 p ISBN:978-3-211-792-70-4). En función del plano de ruptura por el que se produce la ampolla, se definen tres tipos de EB: simple (EBS), juntura! (EBJ), distrófica (EBD) y mixta (EBM). La prevalencia de EB en Europa se estima que es 0,60 por cada 10.000 individuos (Fine et al 2009). Tanto la forma EBD recesiva (EBDR) como la forma EBD dominante (EBDD) son causadas por mutaciones en el gen COL7A1 que codifica para la proteína Colágeno Vil (C7). Esta proteína es el componente principal de las fibrillas de anclaje que engarzan el colágeno de tipo I de la dermis con proteínas de la membrana basal subepidérmica tales como la laminina 332 actuando como un verdadero adhesivo dermo-epidérmico no solo en la piel sino en mucosas como la esofágica. La ausencia o disminución pronunciada de C7 es la característica principal de la EBDR y su determinación establece un diagnóstico molecular primario de la patología. En el desarrollo de la patogénesis de EB se postula una conexión entre el déficit de C7 y una exacerbada biodisponibilidad y sobreexpresión de TGF-bI (Guerra et ai
2017: Guerra L, Gdorisio T, Zambruno G, Castigiia D. Stromai mieroenvironment in type VII collagen-deficient skin: The ground for squamous ceil carcinoma deveiopment. Matrix Biol. 2017 83, 1-10). Análisis proteómicos de muestras de pacientes con EB revelan una elevada expresión en los niveles de varias proteínas de la matriz extracelular como Tenascina C, que alteran la rigidez de la matriz, así como de TNF-alfa e IL-6 relacionados con inflamación mediada por TGF-bI . Estas evidencias indican que el estroma aberrante en EBDR jugaría un papel crítico en la fibrosis y el desarrollo de tumores agresivos y que su normalización sería una vía terapéutica fundamental. El complejo receptor de TGF-bI está formado por el receptor tipo II (TpRIl), el receptor tipo I (ALK5) y el correceptor Endoglina. Epidermoiisis bullosa (EB; OMIM: 120120) comprises a clinically and genetically heterogeneous group of rare skin diseases characterized by the formation of skin blisters, spontaneous or induced by minimal trauma (Fine el al 2009: Fine, Jo-David, Hintner, Helmut. Life with Epidermolysis Bullosa (EB): Etiology, Diagnosis, Multidiscipiinary Care and Therapy. SpringerWienNewYork. 2009, 338 p ISBN: 978-3-211-792-70-4). Depending on the rupture plane through which the blister is produced, three types of EB are defined: simple (EBS), joint! (EBJ), dystrophic (EBD) and mixed (EBM). The prevalence of EB in Europe is estimated to be 0.60 per 10,000 individuals (Fine et al 2009). Both the recessive EBD form (EBDR) and the dominant EBD form (EBDD) are caused by mutations in the COL7A1 gene encoding the Collagen Vil (C7) protein. This protein is the main component of the anchoring fibrils that link type I collagen of the dermis with subepidermal basement membrane proteins such as laminin 332 acting as a true dermo-epidermal adhesive not only on the skin but also on mucosae such as the esophageal. The absence or pronounced decrease of C7 is the main characteristic of EBDR and its determination establishes a primary molecular diagnosis of the pathology. In the development of EB pathogenesis, a connection between C7 deficiency and an exacerbated bioavailability and overexpression of TGF-bI is postulated (Guerra et ai 2017: Guerra L, Gdorisio T, Zambruno G, Castigiia D. Stromai mieroenvironment in type VII collagen-deficient skin: The ground for squamous ceil carcinoma deveiopment. Matrix Biol. 2017 83, 1-10). Proteomic analyzes of samples from EB patients reveal a high expression in the levels of several extracellular matrix proteins such as Tenascin C, which alter the rigidity of the matrix, as well as TNF-alpha and IL-6 related to TGF-mediated inflammation -bI. These evidences indicate that the aberrant stroma in EBDR would play a critical role in fibrosis and the development of aggressive tumors and that its normalization would be a fundamental therapeutic route. The TGF-bI receptor complex consists of the type II receptor (TpRIl), the type I receptor (ALK5) and the co-receptor Endoglin.
La endoglina modula la activación de la fosforilación de ALK5 por parte del receptor T Ril, tras la unión del ligando TGF-bI (Santibañez y cois 2011 : Santibañez JF, Quintanilia M, Bernabeu G. TQR-b/TQR-b receptor system and its role in physiological and pathological conditions. ClinSci (Lond). 2011 Sep;121(6):233-51). El exceso de producción del ligando TGF-bI , puede ser contrarrestado, por un aumento de endoglina, de manera que se prevenga, de este modo, la fosforilación de los coactivadores Smad2/3 y de sus genes diana implicados en fibrosis (síntesis de colágeno, fibronectina, proteínas de matriz extracelular). Estos procesos dan lugar en fibroblastos a la síntesis de matriz extracelular que puede ser exacerbada en procesos profibróticos (exceso de TGF-bI) como sería el caso de la EBDR. Endoglin modulates the activation of phosphorylation of ALK5 by the T Ril receptor, after binding of the TGF-bI ligand (Santibañez et al 2011: Santibañez JF, Quintanilia M, Bernabeu G. TQR-b / TQR-b receptor system and its role in physiological and pathological conditions. ClinSci (Lond). 2011 Sep; 121 (6): 233-51). The excess production of the TGF-bI ligand can be counteracted by an increase in endoglin, so that the phosphorylation of the Smad2 / 3 coactivators and their target genes involved in fibrosis (collagen synthesis) is prevented. , fibronectin, extracellular matrix proteins). These processes give rise to extracellular matrix synthesis in fibroblasts, which can be exacerbated in profibrotic processes (excess TGF-bI), such as EBDR.
Nystróm (Nystróm et al. 2015: Nystróm A, Thriene K, Mittapalli V, Kern J S, KiristiD, Dengjel J &Brucker-Tuderman L. Losarían ameliorates dystrophic epidermoíysis hullosa and uncovers new disease mechanisms. EMBG Molecular Medicine 2015, 7: 1211-1228.) han demostrado que los ratones modelo de la enfermedad EBDR tienen un exceso de producción de TGF-bI en el suero y, que el tratamiento con losartán disminuye su cantidad aunque los efectos secundarios del Losarían pueden limitar su utilización terapéutica (por ejemplo: anemia; mareos, vértigo; hipotensión; alteración renal, fallo renal; astenia, fatiga; hiperpotasemia, aumento de la urea sanguínea, de la creatinina y del potasio séricos; hipoglucemia). Nystróm (Nystróm et al. 2015: Nystróm A, Thriene K, Mittapalli V, Kern JS, KiristiD, Dengjel J & Brucker-Tuderman L. Losarían ameliorates dystrophic epidermoíysis hullosa and uncovers new disease mechanisms. EMBG Molecular Medicine 2015, 7: 1211-1228 .) have shown that EBDR disease model mice have an excess production of serum TGF-bI, and that treatment with losartan decreases its amount although the side effects of Losarían may limit its therapeutic use (for example: anemia ; dizziness, vertigo; hypotension; renal alteration, renal failure; asthenia, fatigue; hyperkalemia, increased blood urea, serum creatinine and potassium; hypoglycemia).
Los moduladores selectivos de ios receptores estrogénicos tales como raioxifeno (Albiñana et al 2010: Albiñana V, Bernabeu-Herrero ME, Zarrabeitia R, Bernabéu C, Botella LM. Estrogen therapy for hereditary haemorrhagic telangiectasia (HHT): Effects of raloxifene, on Endoglin and ALK1 expression in endothelial cells.
ThrombHaemost 2010 Mar;1G3(3):525~34) y bazedoxifeno (Zarrabeitia et aL, 2016: Zarrabeiíia R, Ojeda-Fernandez L, Recio L, Bernabéu C, Parra JA, Aibiñana V, Botella LM. Bazedoxifene, a new orphan drug for the íreaiment of bleeding in hereditary haemorrhagic telangiectasia. Thromb Haemost. 2016 Jun 2; 115(6): 1167-77), producen un aumento de los niveles proteicos de endoglina y un descenso en los niveles proteicos de TGF-bI en ios modelos de endotelio vascular. Otro compuesto que activa la producción de endoglina e inhibe la producción de TGF-bI es la N- acetiicisteína (NAC). El antioxidante N-acetilcisteína (NAC) tiene un reconocido valor terapéutico en la inflamación, fibrosis, disfunción endoteiial y promueve la síntesis de un gran número de genes mediante la modulación de la señalización de vías de diferentes quinasas, un concepto útil para el tratamiento de ciertas enfermedades degenerativas (Zafaruliah M, Li WQ, Sylvester J, Ahmad M. Molecular mechanisms of N-acetylcysteine actions.Cell Mol Life Sel. 2003 Jan;60(1):6~20) Selective modulators of estrogenic receptors such as raioxifene (Albiñana et al 2010: Albiñana V, Bernabeu-Herrero ME, Zarrabeitia R, Bernabéu C, Botella LM. Estrogen therapy for hereditary haemorrhagic telangiectasia (HHT): Effects of raloxifene, on Endoglin ALK1 expression in endothelial cells. ThrombHaemost 2010 Mar; 1G3 (3): 525 ~ 34) and bazedoxifene (Zarrabeitia et aL, 2016: Zarrabeiíia R, Ojeda-Fernandez L, Recio L, Bernabéu C, Parra JA, Aibiñana V, Botella LM. Bazedoxifene, a new orphan drug for the íreaiment of bleeding in hereditary haemorrhagic telangiectasia. Thromb Haemost. 2016 Jun 2; 115 (6): 1167-77), cause an increase in endoglin protein levels and a decrease in TGF-bI protein levels in ios vascular endothelial models. Another compound that activates endoglin production and inhibits TGF-bI production is N-acetiicysteine (NAC). The antioxidant N-acetylcysteine (NAC) has a recognized therapeutic value in inflammation, fibrosis, endoteiial dysfunction and promotes the synthesis of a large number of genes by modulating signaling pathways of different kinases, a useful concept for the treatment of certain degenerative diseases (Zafaruliah M, Li WQ, Sylvester J, Ahmad M. Molecular mechanisms of N-acetylcysteine actions.Cell Mol Life Sel. 2003 Jan; 60 (1): 6 ~ 20)
Por tanto, sería deseable disponer de fármacos que sean eficaces y selectivos para el tratamiento de las fibrosis y en particular de una enfermedad rara como la epidermólisis hullosa (EB) que supongan un aumento en la calidad de vida de los pacientes de estas enfermedades. Therefore, it would be desirable to have drugs that are effective and selective for the treatment of fibrosis and in particular of a rare disease such as epidermolysis hullosa (EB) that represent an increase in the quality of life of patients with these diseases.
DESCRIPCIÓN DE LA INVENCIÓN DESCRIPTION OF THE INVENTION
En un primer aspecto, la presente invención se refiere a un compuesto caracterizado por que es inhibidor de la ruta de TGF-bI y estimulador de endoglina para su uso en el tratamiento de genodermatosis. In a first aspect, the present invention relates to a compound characterized in that it is an inhibitor of the TGF-bI pathway and an endoglin stimulator for use in the treatment of genodermatosis.
En otra realización la invención se refiere al compuesto según el uso definido anteriormente seleccionado de raloxifeno y bazedoxifeno. In another embodiment the invention relates to the compound according to the previously defined use selected for raloxifene and bazedoxifene.
En otra realización la invención se refiere al compuesto según el uso definido anteriormente, donde la enfermedad se selecciona de epidermólisis hullosa (EB). In another embodiment the invention relates to the compound according to the use defined above, where the disease is selected from epidermolysis hullosa (EB).
En otra realización la invención se refiere al compuesto según el uso definido anteriormente, donde la EB se selecciona de EB simple (EBS), EB juntural (EBJ), EB distrófica (EBD) y síndrome de Kindler
En otra realización la invención se refiere al compuesto según el uso definido anteriormente, donde la EBD se selecciona de EBD recesiva (EBDR) y EBD dominante (EBDD). In another embodiment the invention relates to the compound according to the use defined above, where EB is selected from simple EB (EBS), junctional EB (EBJ), dystrophic EB (EBD) and Kindler's syndrome In another embodiment the invention relates to the compound according to the use defined above, where EBD is selected from recessive EBD (EBDR) and dominant EBD (EBDD).
En otra realización la invención se refiere al compuesto según el uso definido anteriormente, donde el compuesto se selecciona de raloxifeno, bazedoxifeno y N- acetiicisteína, preferiblemente donde el compuesto raloxifeno y N-acetiicisteína, para el tratamiento de genodermatosis, preferiblemente para el tratamiento de epidermólisis hullosa (EB), más preferiblemente para el tratamiento de EB simple (BBS), EB juntural (EBJ), EB distrófica (EBD) o síndrome de Kindier y aún más preferiblemente para el tratamiento de EBD recesiva (EBDR) o EBD dominante (EBDD). In another embodiment the invention relates to the compound according to the use defined above, where the compound is selected from raloxifene, bazedoxifene and N-acetiicysteine, preferably where the compound raloxifene and N-acetiicysteine, for the treatment of genodermatosis, preferably for the treatment of epidermolysis hullosa (EB), more preferably for the treatment of simple EB (BBS), junctional EB (EBJ), dystrophic EB (EBD) or Kindier syndrome and even more preferably for the treatment of recessive EBD (EBDR) or dominant EBD ( EBDD).
Otro aspecto de ¡a presente invención se refiere a una composición farmacéutica que comprende uno o más compuestos caracterizados por que son inhibidores de la ruta de TGF-bI y estimuladores de endogiina para su uso en ei tratamiento de genodermatosis y uno o más excipientes farmacéuticamente aceptables para su uso definido anteriormente. Another aspect of the present invention relates to a pharmaceutical composition comprising one or more compounds characterized in that they are inhibitors of the TGF-bI pathway and endogiine stimulators for use in the treatment of genodermatosis and one or more pharmaceutically acceptable excipients. for its previously defined use.
En otra realización la invención se refiere a la composición farmacéutica definida anteriormente que comprende raloxifeno y N-acetilcisteína. In another embodiment the invention relates to the pharmaceutical composition defined above comprising raloxifene and N-acetylcysteine.
En otra realización la invención se refiere a la composición farmacéutica definida anteriormente que comprende bazedoxifeno y N-acetilcisteína. In another embodiment the invention relates to the pharmaceutical composition defined above comprising bazedoxifene and N-acetylcysteine.
Así pues, la presente invención demuestra que el tratamiento de ios fibroblastos de pacientes de EBDR con Raloxifeno y NAC provoca la sobreexpresión de endogiina y, podría está asociado con la represión observada de ios niveles de TGF-bI y de sus dianas de matriz celular. En este sentido, los resultados muestran que ios niveles de la forma activa de TGF-bI en el suero de los pacientes de EB probados es un biomarcador de la enfermedad. Ai disminuir la expresión de TGF-bI activo, ios genes de matriz extraceluiar que regula (colágeno tipo 1 , tenascina, decorina, fibronectina, periostina, TGF-bI) son reprimidos y ia matriz extraceluiar se normalizaría. Thus, the present invention demonstrates that the treatment of fibroblasts from EBDR patients with Raloxifene and NAC causes the overexpression of endogiin and could be associated with the observed repression of the levels of TGF-bI and its targets of cellular matrix. In this sense, the results show that the levels of the active form of TGF-bI in the serum of the EB patients tested is a biomarker of the disease. By decreasing the expression of active TGF-bI, the extracellular matrix genes it regulates (type 1 collagen, tenascin, decorin, fibronectin, periostin, TGF-bI) are repressed and the extracellular matrix would normalize.
Este mecanismo farmacológico de represión de ios niveles de TGF-bI y ia consiguiente regulación de las proteínas de matriz celular, justifica ia protección de
estos fármacos en ¡os procesos fisiopatológicos de fibrosis. This pharmacological mechanism of repression of TGF-bI levels and the consequent regulation of cellular matrix proteins, justifies the protection of these drugs in the pathophysiological processes of fibrosis.
A ¡o ¡argo de la descripción y las reivindicaciones ¡a palabra "comprende" y sus variantes no pretenden excluir otras características técnicas, aditivos, componentes o pasos. Para ios expertos en ia materia, otros objetos, ventajas y características de ¡a invención se desprenderán en parte de la descripción y en parte de ¡a práctica de ¡a invención. Los siguientes ejemplos y figuras se proporcionan a modo de ilustración, y no se pretende que sean ¡imitativos de ¡a presente invención. Through the description and claims the word "comprises" and its variants are not intended to exclude other technical characteristics, additives, components or steps. For those skilled in the art, other objects, advantages, and characteristics of the invention will emerge in part from the description and in part from the practice of the invention. The following examples and figures are provided by way of illustration, and are not intended to be imitative of the present invention.
BREVE DESCRIPCIÓN DE LAS FIGURAS BRIEF DESCRIPTION OF THE FIGURES
FIG. 1 Determinación de ¡os niveles de TGF-bI activo en 23 sueros de pacientes con Epidermóüsis Bullosa Recesiva Dominante (EBDR). Niveles de TGF-bI activo (A) y total (B) en sujetos sanos y pacientes EBDR. FIG. 1 Determination of the levels of active TGF-bI in 23 sera from patients with Dominant Recessive Epidermolysis Bullosa (EBDR). Active (A) and total (B) TGF-bI levels in healthy subjects and EBDR patients.
FIG, 2 Raloxifeno y NAC modulan los niveles de proteína de endogfína y de IGF- b1. El efecto del raloxifeno (0,2hM) and NAC (HQQuIVI} en ios niveles de endoglina y TGF-bI activo y total fue analizado mediante ELISA bajo condiciones de deprivación de suero y en presencia de¡ fármaco, durante 48h, mediante ELISA. Las figuras 2A, 2C y 2E representan la ratio de cambio (fold change) en los niveles de endoglina y TGF en los fibroblastos de 3 pacientes, Las figuras 2B, 2D y 2F representan la media de dichos cambios en dichos 3 pacientes, *p<0,Q5; **p<0,001 ; ***p<0,001. FIG, 2 Raloxifene and NAC modulate levels of endogfin protein and IGF-b1. The effect of raloxifene (0.2hM) and NAC (HQQuIVI} on endoglin levels and total and active TGF-bI was analyzed by ELISA under serum deprivation conditions and in the presence of the drug, during 48h, by ELISA. Figures 2A, 2C and 2E represent the rate of change (fold change) in the levels of endoglin and TGF in the fibroblasts of 3 patients. Figures 2B, 2D and 2F represent the mean of these changes in said 3 patients, * p < 0, Q5; ** p <0.001; *** p <0.001.
FIG. 3 El raloxifeno y el NAC regulan expresión a nivel mRNA de proteínas asociadas a fibrosis. (A) se midieron ¡os niveles de RNA de endoglina y TGF-b en fibroblastos de pacientes cultivados en presencia o ausencia de raloxifeno (0,2nM) o NAC (100uM) durante 48h (B) Niveles de expresión de proteínas asociadas a fibrosis (Tenascina C, Periostina, TGFi, TSP y !L-6) medidas por PCR cuantitativa real (qRT- PCR) en ¡as condiciones anteriores. Los niveles de tránscrito fueron normalizados con el control de RNA ribosómico ACTB, *p<Q,05; **p<0,001 ; ***p<0,001. FIG. 3 Raloxifene and NAC regulate mRNA expression of proteins associated with fibrosis. (A) Endoglin and TGF-b RNA levels were measured in fibroblasts from patients cultured in the presence or absence of raloxifene (0.2nM) or NAC (100uM) for 48h (B) Expression levels of proteins associated with fibrosis (Tenascin C, Periostin, TGFi, TSP and! L-6) measured by real quantitative PCR (qRT-PCR) under the above conditions. Transcript levels were normalized with the ACTB ribosomal RNA control, * p <Q, 05; ** p <0.001; *** p <0.001.
FIG. 4 Ensayo de capacidad contráctil de fibroblastos de 1 paciente EBDR en geies de
colágeno en ausencia y presencia de raloxifeno y iosartán. FIG. 4 Test of contractile capacity of fibroblasts from 1 EBDR patient in geys of collagen in the absence and presence of raloxifene and iosartan.
FSG. 5 Ensayo de capacidad contráctil de fibroblastos de 4 pacientes de la cohorte EBDR en geies de colágeno en ausencia y presencia de NAC. FSG. 5 Test of contractile capacity of fibroblasts from 4 patients of the EBDR cohort in collagen geies in the absence and presence of NAC.
FIG, 8 Mecanismos de los fármacos en estudio en fibroblastos de pacientes de EBDR, sobre la ruta de señalización de TGF-bI , mediante el análisis de la expresión de ias proteínas Smad2 y Smad 1/5 fosforiladas. Fibroblastos humanos de pacientes con EBDR fueron transinfectados con los vectores promotor-reporte CAGA-luc (A y B) y BRE-luc (C y D). La actividad iuciferasa fue medida en las células sin tratar o tratadas con raloxifeno o NAC durante 48b y en la presencia de TGF-b durante las últimas 24b (E) o 3h (F). Se muestran WB representativos. Las figuras A y C representa resultados individuales de 3 pacientes, y B y D la ratio de inducción (fold induction) de ios mismos. Los ensayos de iuciferasa se realizaron como se indica en Material y Métodos. Como control de carga se empleó Smad2 o Smad1/5 total, la b-actina muestra también un resultado similar. Las diferencias son estadísticamente significativas de acuerdo con el test de Student *P < 0,05; **P < 0,01 ; ***P < 0,005. FIG, 8 Mechanisms of drugs under study in fibroblasts from EBDR patients, on the TGF-bI signaling pathway, by analyzing the expression of phosphorylated Smad2 and Smad 1/5 proteins. Human fibroblasts from patients with EBDR were transfected with the CAGA-luc (A and B) and BRE-luc (C and D) promoter-report vectors. Iuciferase activity was measured in untreated or raloxifene or NAC-treated cells for 48b and in the presence of TGF-b for the last 24b (E) or 3h (F). Representative WBs are shown. Figures A and C represent individual results from 3 patients, and B and D the fold induction ratio of themselves. Iuciferase assays were performed as indicated in Material and Methods. Smad2 or total Smad1 / 5 was used as loading control, b-actin also shows a similar result. The differences are statistically significant according to the Student test * P <0.05; ** P <0.01; *** P <0.005.
EJEMPLOS EXAMPLES
A continuación, se ilustrará la invención mediante unos ensayos realizados por ios inventores, que pone de manifiesto la efectividad del compuesto de la invención. Next, the invention will be illustrated by means of tests carried out by the inventors, which shows the effectiveness of the compound of the invention.
Ejemplo 1 : Determinación de ios niveles de TGF-bI activo en 23 sueros de pacientes con Epider ólisis buliosa. Example 1: Determination of ios levels of active TGF-bI in 23 sera from patients with Epiderolysis buliosa.
La disponibilidad de un total de 23 sueros de pacientes de EBDR seleccionados por edad (entre 18-40 años) y por ser portadores de idéntica mutación en el C7 mutación c.6527insC en el exón 80 del gen COL7A 1, ba permitido la determinación de ios niveles de TGF-bI . La concentración del TGF-bI se determinó mediante inmunoensayo enzimático (ELISA; R & D Systems). Los niveles de TGF-bI fueron determinados tanto en sueros no acidificados (TGF-bI -total) como acidificados (TGF- bΐ-activo). En la Figura 1A se puede observar la elevación de los niveles de TGF-bI activo en los sueros de los pacientes de EBDR, frente a la prácticamente ausencia (detectabie) de la proteína activa en los controles sanos. La Figura 1 B muestra ios
niveles de TGF-bI total en los sueros de las muestras analizadas. The availability of a total of 23 sera from EBDR patients selected by age (between 18-40 years) and because they carry the same mutation in the C7 mutation c.6527insC in exon 80 of the COL7A 1 gene, allowed the determination of TGF-bI levels. TGF-bI concentration was determined by enzyme immunoassay (ELISA; R&D Systems). TGF-bI levels were determined in both non-acidified (TGF-bI -total) and acidified (TGF-bΐ-active) sera. Figure 1A shows the elevation of the levels of active TGF-bI in the sera of EBDR patients, compared to the practically absence (detectability) of the active protein in healthy controls. Figure 1 B shows ios total TGF-bI levels in the sera of the samples analyzed.
Este resultado parecería señalar a la forma activa de TGF-bI como un potencial biomarcador diagnóstico y/o de seguimiento y/o pronóstico de la enfermedad, lo que indudablemente tendría una gran trascendencia clínica. This result would seem to point to the active form of TGF-bI as a potential biomarker for diagnosis and / or monitoring and / or prognosis of the disease, which undoubtedly would have great clinical significance.
Ejemplo 2: Efecto farmacológico del raloxifeno, la N-acetilclstefna (NAC) y el losartán en fibroblastos de pacientes de EBDR. Example 2: Pharmacological effect of raloxifene, N-acetylclstefna (NAC) and losartan in fibroblasts from EBDR patients.
La Figura 2 (paneles A-C) muestra ios niveles detectados de proteínas Endoglina y TGF-bI activo y TGF-bI total, respectivamente, en ios fibroblastos de tres pacientes EBDR tratados con el raloxifeno, la N-acetilcisteína (NAC) y el losartán. Mientras que los niveles de TGF-bI total presentan respuestas diversas a los tratamientos según el paciente, la Figura 2B muestra la reducción significativa de los niveles de TGF-bI activo con los tres tratamientos farmacológicos. Esta disminución en los niveles de la citoquina activa es especialmente conspicua con el tratamiento con el raloxifeno en ios fibroblastos de los pacientes 17 y 31 y con la NAC en el paciente 8. El losartán se presenta especialmente activo en el paciente 31 que parece el más sensible a cualquiera de ios tratamientos efectuados. Figure 2 (panels A-C) shows the detected levels of Endoglin proteins and active TGF-bI and total TGF-bI, respectively, in the fibroblasts of three EBDR patients treated with raloxifene, N-acetylcysteine (NAC) and losartan. While the total TGF-bI levels show different responses to the treatments according to the patient, Figure 2B shows the significant reduction in the levels of active TGF-bI with the three pharmacological treatments. This decrease in the levels of the active cytokine is especially conspicuous with the treatment with raloxifene in the fibroblasts of patients 17 and 31 and with CAP in patient 8. Losartan appears especially active in patient 31, who seems to be the most sensitive to any of the treatments carried out.
El tratamiento con estos fármacos (raloxifeno, NAC y losartán) lleva a la elevación de ios niveles de endoglina y a la reducción de la expresión de! TGF-bI activo, en rangos que oscilan entre el 7G%-90% con respecto a los controles. Treatment with these drugs (raloxifene, NAC, and losartan) leads to an increase in endoglin levels and a reduction in the expression of! Active TGF-bI, in ranges ranging from 7G% -90% with respect to controls.
No está descrito que el losartán aumente los niveles de expresión de endoglina ni que raloxifeno y NAC modulen ios niveles de TGF-bI activo en fibroblastos humanos. Losartan has not been reported to increase endoglin expression levels or that raloxifene and NAC modulate levels of active TGF-bI in human fibroblasts.
Ejemplo 3: El raloxifeno, la NAC y el losartán reprimen los niveles transcripcionales de las proteínas de matriz Tenascina C, Periostina, y TGFi en fibroblastos de pacientes de EBDR, Example 3: Raloxifene, NAC, and losartan suppress transcriptional levels of matrix proteins Tenascin C, Periostin, and TGFi in fibroblasts from EBDR patients,
El desarrollo de la patogénesis de EBDR se postula una conexión entre el déficit de C7 y una exacerbada biodisponibilidad y sobreexpresión de TGF-bI y de los genes regulados por este factor, que incluyen la expresión de proteínas de matriz celular.
Los análisis proteómicos de muestras de pacientes con EBDR revelan una elevada expresión en los niveles de varias proteínas relacionadas con inflamación mediada por TGF-bI y de proteínas de la matriz extracelular como Tenascina C (Guerra et al 2017). La Figura 3 muestra la disminución de los niveles de mRNA por determinaciones de RT-PCR de las proteínas de matriz celular Tenascina C, Periostina, TGFi y Tromboespondina-1 (TSP1) así como de la de la citoquina marcadora de inflamación IL-6. The development of EBDR pathogenesis postulates a connection between C7 deficiency and exacerbated bioavailability and overexpression of TGF-bI and genes regulated by this factor, including expression of cellular matrix proteins. Proteomic analyzes of samples from patients with EBDR reveal a high expression in the levels of several proteins related to inflammation mediated by TGF-bI and of proteins of the extracellular matrix such as Tenascin C (Guerra et al 2017). Figure 3 shows the decrease in mRNA levels by RT-PCR determinations of the cellular matrix proteins Tenascin C, Periostin, TGFi and Thromboespondin-1 (TSP1) as well as that of the inflammatory marker cytokine IL-6.
Ejemplo 4: El raloxifeno inhibe la contracción de la superficie de proliferación de los fibroblastos de un paciente de EBDR en geles de colágeno. Example 4: Raloxifene inhibits fibroblast proliferation surface contraction of an EBDR patient in collagen gels.
El ensayo de contracción de colágeno (Ferrer-Mayorga et ai 2017: Ferrer-Mayorga G, et a!. Vitamin D receptor expression and associated gene signature in tumour stromal fibrobiasts predict clinicai outcome in colorectai cáncer, Gut 2017, 66:1449- 1462) se utiliza para explorar la activación de los fibroblastos y consiguiente reducción de la superficie ocupada por las células en cultivo, en ausencia de adhesión de a ios soportes de las paredes de la placa de ensayo. El raloxifeno inhibe una de las propiedades profibróticas como es la contracción de las fibras de colágeno en gel (ver Figura 4), lo que constituye una característica ilustrativa de la activación de ios fibroblastos. The collagen contraction test (Ferrer-Mayorga et ai 2017: Ferrer-Mayorga G, et a !. Vitamin D receptor expression and associated gene signature in tumor stromal fibrobiasts predict clinicai outcome in colorectai cancer, Gut 2017, 66: 1449-1462 ) is used to explore the activation of fibroblasts and consequent reduction of the surface occupied by cells in culture, in the absence of adhesion to the supports of the walls of the assay plate. Raloxifene inhibits one of the profibrotic properties, such as the contraction of collagen gel fibers (see Figure 4), which is an illustrative characteristic of the activation of fibroblasts.
Ejemplo 5: La ISI-acetilcisteína inhibe la contracción de las fibras de colágeno de los fibroblastos en geles de colágeno de cuatro pacientes de EBDR. Example 5: ISI-acetylcysteine inhibits the collagen fibers contraction of fibroblasts in collagen gels from four EBDR patients.
El ensayo de contracción de colágeno (Ferrer-Mayorga et al 2017) se utiliza para explorar la activación de ios fibroblastos por la reducción de la superficie ocupada por las células en cultivo, en ausencia de adhesión a las paredes de la placa de ensayo (ver panel control en Figura 5). La N-acetilcisteína inhibe una de las propiedades profibróticas como es ía contracción de las fibras de colágeno en gel, ios que constituye una característica ilustrativa de ía activación de ios fibroblastos (Figura 5). The collagen contraction assay (Ferrer-Mayorga et al 2017) is used to explore the activation of ios fibroblasts by reducing the surface occupied by cells in culture, in the absence of adhesion to the walls of the assay plate (see control panel in Figure 5). N-acetylcysteine inhibits one of the profibrotic properties such as the contraction of the collagen fibers in gel, which is an illustrative characteristic of the activation of the fibroblasts (Figure 5).
Ejemplo 6: El raloxifeno y la MAC inhiben ios niveles de fosforilación de Smad2/3 y Smad 1/5 fosforiiados. Example 6: Raloxifene and MAC inhibit phosphorylation levels of Smad2 / 3 and Smad 1/5 phosphorylated.
La disminución de producción del ligando TGF-bI , asociado al aumento de endogíina (Ejemplo 2) de manera que se prevenga, de este modo, la fosforilación de ios
coactivadores de la ruta SMAD y, diana implicadas en fibrosis (síntesis de colágeno, fibronectina, proteínas de matriz extracelular). Estos procesos dan lugar en células vasculares a la estabilización de los vasos, y en fibroblastos a la síntesis de matriz extracelular que puede ser exacerbada en procesos profibróticos The decrease in production of the TGF-bI ligand, associated with the increase in endogyny (Example 2) so as to prevent phosphorylation of ios co-activators of the SMAD and target pathway implicated in fibrosis (synthesis of collagen, fibronectin, extracellular matrix proteins). These processes lead to vascular stabilization in vascular cells, and in fibroblasts to extracellular matrix synthesis that can be exacerbated in profibrotic processes.
Los experimentos con fibroblastos de pacientes de EBDR transfectados transitoriamente tanto con reporteros de Smad 2/3 (CAGA-LUC) como de Smad 1/5 (BRE-LUC), en ausencia y presencia de ios fármacos en estudio, demostraron una actividad inhibitoria de todos ellos tanto en la ruta de Smad2/3 como en 1/5 (Figura 6) Experiments with fibroblasts from EBDR patients transiently transfected with both Smad 2/3 (CAGA-LUC) and Smad 1/5 (BRE-LUC) reporters, in the absence and presence of the study drugs, demonstrated an inhibitory activity of all of them both in the Smad2 / 3 route and in 1/5 (Figure 6)
El raloxifeno y la NAC parecen afectar a la ruta de señalización TGF-b /Smad de forma diferente ai Losarían, según ios resultados de Nystrom y cois): mientras que el raloxifeno parece inhibir la ruta Smad2/3 de forma selectiva, ia NAC inhibe indiscriminadamente tanto ambas rutas, Smad 2/3 y Smad 1/5.
Raloxifene and NAC appear to affect the TGF-b / Smad signaling pathway differently than Losarían, according to the results of Nystrom et al.): While raloxifene appears to selectively inhibit the Smad2 / 3 pathway, ia NAC inhibits Both routes indiscriminately, Smad 2/3 and Smad 1/5.
Claims
1. Compuesto caracterizado por que es inhibidor de la ruta de TGF-bI y estimulador de endoglina para su uso en el tratamiento de genodermatosis 1. Compound characterized in that it is an inhibitor of the TGF-bI pathway and an endoglin stimulator for use in the treatment of genodermatosis
2. El compuesto según el uso de ¡a reivindicación 1 , donde e¡ compuesto se selecciona de raloxifeno y bazedoxifeno. 2. The compound according to the use of claim 1, wherein the compound is selected from raloxifene and bazedoxifene.
3. El compuesto según el uso de la reivindicación 2, donde el compuesto es raloxifeno 3. The compound according to the use of claim 2, wherein the compound is raloxifene
4. El compuesto según el uso de cualquiera de las reivindicaciones 1 a 3, donde la genodermatosis se selecciona de epidermólisis hullosa (EB) 4. The compound according to the use of any of claims 1 to 3, wherein the genodermatosis is selected from epidermolysis hullosa (EB)
5. El compuesto según el uso de la reivindicación 4, donde la EB se selecciona de EB simple (BBS), EB juntural (EBJ), EB distrófica (EBD) y síndrome de Kindler 5. The compound according to the use of claim 4, wherein the EB is selected from simple EB (BBS), junctional EB (EBJ), dystrophic EB (EBD) and Kindler's syndrome
6. El compuesto según el uso de la reivindicación 5, donde la EBD se selecciona de EBD recesiva (EBDR) y EBD dominante (EBDD). 6. The compound according to the use of claim 5, wherein the EBD is selected from recessive EBD (EBDR) and dominant EBD (EBDD).
7. Una composición farmacéutica que comprende uno o más compuestos caracterizados por que son inhibidores de la ruta de TGF-bI y estimuladores de endoglina para su uso en el tratamiento de genodermatosis y uno o más excipientes farmacéuticamente aceptables para su uso según cualquiera de las reivindicaciones 1 a 6. 7. A pharmaceutical composition comprising one or more compounds characterized in that they are TGF-bI pathway inhibitors and endoglin stimulators for use in the treatment of genodermatosis and one or more pharmaceutically acceptable excipients for use according to any of the claims 1 to 6.
8. La composición según la reivindicación 7 que comprende raloxifeno y N- acetiicisteína 8. The composition according to claim 7 comprising raloxifene and N-acetiicysteine
9. La composición según la reivindicación 7, que comprende bazedoxifeno y N~ acetilcisteína
9. The composition according to claim 7, comprising bazedoxifene and N ~ acetylcysteine
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
ESP201831072 | 2018-11-07 | ||
ES201831072A ES2759400A1 (en) | 2018-11-07 | 2018-11-07 | TGF-ß1 INHIBITORS AND ENDOGLIN PRODUCERS FOR USE IN THE TREATMENT OF BULLYUS EPIDERMOLISIS (Machine-translation by Google Translate, not legally binding) |
Publications (1)
Publication Number | Publication Date |
---|---|
WO2020094902A1 true WO2020094902A1 (en) | 2020-05-14 |
Family
ID=70483298
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/ES2019/070757 WO2020094902A1 (en) | 2018-11-07 | 2019-11-07 | Inhibitors of tgf-β1 and endoglin producers for use in the treatment of epidermolysis bullosa |
Country Status (2)
Country | Link |
---|---|
ES (1) | ES2759400A1 (en) |
WO (1) | WO2020094902A1 (en) |
Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20030229141A1 (en) * | 1999-01-08 | 2003-12-11 | Yu Ruey J. | N-acetyl cysteine and its topical use |
WO2013138744A1 (en) * | 2012-03-16 | 2013-09-19 | M. Alphabet 1, Llc | Compositions for the treatment of skin disorders |
US20180015060A1 (en) * | 2015-01-27 | 2018-01-18 | Florengale, Llc | Healing topical composition |
-
2018
- 2018-11-07 ES ES201831072A patent/ES2759400A1/en active Pending
-
2019
- 2019-11-07 WO PCT/ES2019/070757 patent/WO2020094902A1/en active Application Filing
Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20030229141A1 (en) * | 1999-01-08 | 2003-12-11 | Yu Ruey J. | N-acetyl cysteine and its topical use |
WO2013138744A1 (en) * | 2012-03-16 | 2013-09-19 | M. Alphabet 1, Llc | Compositions for the treatment of skin disorders |
US20180015060A1 (en) * | 2015-01-27 | 2018-01-18 | Florengale, Llc | Healing topical composition |
Non-Patent Citations (2)
Title |
---|
ALBIÑANA VIRGINIA ET AL.: "Estrogen therapy for hereditary haemorrhagic telangiectasia (HHT): Effects of raloxifene, on Endoglin and ALK1 expression in endothelial cells", THROMBOSIS AND HAEMOSTASIS MAR 2010, vol. 103, no. 3, 28 February 2010 (2010-02-28), pages 525 - 534, XP055707254, ISSN: 0340-6245 * |
OZMEN BILGIN ET AL.: "Influence of the selective oestrogen receptor modulator (raloxifene hydrochloride) on IL -6, TNF-alpha, TGF-beta 1 and bone turnover markers in the treatment of postmenopausal osteoporosis", EUROPEAN CYTOKINE NETWORK SEP 2007, vol. 18, no. 3, 31 August 2007 (2007-08-31), pages 148 - 153, XP055707251, ISSN: 1952-4005 * |
Also Published As
Publication number | Publication date |
---|---|
ES2759400A1 (en) | 2020-05-08 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
Neufeld et al. | The role of the semaphorins in cancer | |
Galiè | RAS as supporting actor in breast cancer | |
Jadaliha et al. | Functional and prognostic significance of long non-coding RNA MALAT1 as a metastasis driver in ER negative lymph node negative breast cancer | |
Zhou et al. | Proteomics identification of annexin A2 as a key mediator in the metastasis and proangiogenesis of endometrial cells in human adenomyosis | |
Ali et al. | Expression and mutation analysis of epidermal growth factor receptor in head and neck squamous cell carcinoma | |
Paranjpe et al. | RNA interference against hepatic epidermal growth factor receptor has suppressive effects on liver regeneration in rats | |
Lay et al. | Interleukin 11 regulates endometrial cancer cell adhesion and migration via STAT3 | |
ES2727904T3 (en) | Method for the prognosis and treatment of metastatic bone cancer that originates from breast cancer | |
Koumangoye et al. | Reduced annexin A6 expression promotes the degradation of activated epidermal growth factor receptor and sensitizes invasive breast cancer cells to EGFR-targeted tyrosine kinase inhibitors | |
Rea et al. | Novel Axl-driven signaling pathway and molecular signature characterize high-grade ovarian cancer patients with poor clinical outcome | |
Corachán et al. | Vitamin D as an effective treatment in human uterine leiomyomas independent of mediator complex subunit 12 mutation | |
Sulzmaier et al. | RSK2 activity mediates glioblastoma invasiveness and is a potential target for new therapeutics | |
Pehkonen et al. | Liprin-α1 modulates cancer cell signaling by transmembrane protein CD82 in adhesive membrane domains linked to cytoskeleton | |
Li et al. | Novel role of semaphorin 3A in the growth and progression of hepatocellular carcinoma | |
Xiao et al. | ELF1 activated long non-coding RNA CASC2 inhibits cisplatin resistance of non-small cell lung cancer via the miR-18a/IRF-2 signaling pathway. | |
Zhao et al. | A positive feedback loop of miR-30a-5p-WWP1-NF-κB in the regulation of glioma development | |
Qu et al. | Osterix promotes the migration and angiogenesis of breast cancer by upregulation of S100A4 expression | |
Ng et al. | Krüppel-like factor 5 promotes sonic hedgehog signaling and neoplasia in Barrett's esophagus and esophageal adenocarcinoma | |
Sakai et al. | MET increases the sensitivity of gefitinib-resistant cells to SN-38, an active metabolite of irinotecan, by up-regulating the topoisomerase I activity | |
Sung et al. | EGFL6 promotes colorectal cancer cell growth and mobility and the anti‐cancer property of anti-EGFL6 antibody | |
Chen et al. | Knockdown of Parkinson’s disease-related gene ATP13A2 reduces tumorigenesis via blocking autophagic flux in colon cancer | |
Han et al. | HIF-1α induced NID1 expression promotes pulmonary metastases via the PI3K-AKT pathway in salivary gland adenoid cystic carcinoma | |
WO2020094902A1 (en) | Inhibitors of tgf-β1 and endoglin producers for use in the treatment of epidermolysis bullosa | |
Ellrichmann et al. | Gastrin stimulates the VEGF-A promotor in a human colon cancer cell line | |
Tu et al. | Secreted phosphoprotein 1 promotes angiogenesis of glioblastoma through upregulating PSMA expression via transcription factor HIF1α: SPP1 promotes angiogenesis of glioblastoma |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
121 | Ep: the epo has been informed by wipo that ep was designated in this application |
Ref document number: 19881961 Country of ref document: EP Kind code of ref document: A1 |
|
NENP | Non-entry into the national phase |
Ref country code: DE |
|
122 | Ep: pct application non-entry in european phase |
Ref document number: 19881961 Country of ref document: EP Kind code of ref document: A1 |