[go: up one dir, main page]

WO2018147615A1 - Composition pharmaceutique pour inhibition de l'angiogenèse contenant un penta-depsipeptide cyclique en tant que principe actif - Google Patents

Composition pharmaceutique pour inhibition de l'angiogenèse contenant un penta-depsipeptide cyclique en tant que principe actif Download PDF

Info

Publication number
WO2018147615A1
WO2018147615A1 PCT/KR2018/001560 KR2018001560W WO2018147615A1 WO 2018147615 A1 WO2018147615 A1 WO 2018147615A1 KR 2018001560 W KR2018001560 W KR 2018001560W WO 2018147615 A1 WO2018147615 A1 WO 2018147615A1
Authority
WO
WIPO (PCT)
Prior art keywords
angiogenesis
formula
cyclic
pharmaceutical composition
pentapsipeptide
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
PCT/KR2018/001560
Other languages
English (en)
Korean (ko)
Inventor
이찬
문성권
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Industry Academic Cooperation Foundation of Chung Ang University
Original Assignee
Industry Academic Cooperation Foundation of Chung Ang University
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Priority claimed from KR1020180011886A external-priority patent/KR102022631B1/ko
Application filed by Industry Academic Cooperation Foundation of Chung Ang University filed Critical Industry Academic Cooperation Foundation of Chung Ang University
Priority to US16/320,502 priority Critical patent/US10709757B2/en
Priority to CN201880002835.7A priority patent/CN109689083B/zh
Publication of WO2018147615A1 publication Critical patent/WO2018147615A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

Links

Images

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/04—Peptides having up to 20 amino acids in a fully defined sequence; Derivatives thereof
    • A61K38/08—Peptides having 5 to 11 amino acids
    • 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
    • A61P17/06—Antipsoriatics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/04—Peptides having up to 20 amino acids in a fully defined sequence; Derivatives thereof
    • A61K38/12—Cyclic peptides, e.g. bacitracins; Polymyxins; Gramicidins S, C; Tyrocidins A, B or C
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/04—Peptides having up to 20 amino acids in a fully defined sequence; Derivatives thereof
    • A61K38/15—Depsipeptides; Derivatives thereof
    • 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
    • A61P17/10—Anti-acne agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P19/00—Drugs for skeletal disorders
    • A61P19/02—Drugs for skeletal disorders for joint disorders, e.g. arthritis, arthrosis
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P27/00—Drugs for disorders of the senses
    • A61P27/02—Ophthalmic agents

Definitions

  • the present invention relates to a pharmaceutical composition for inhibiting angiogenesis (NeoN-methylsansalvamide, NMSSV) containing an cyclic pentapsipeptide peptide (NeoN-methylsansalvamide, NMSSV) as an active ingredient.
  • Angiogenesis is a biological process that creates new blood vessels into tissues or organs. Under normal physiological conditions, humans or animals produce neovascularization only in very limited circumstances.
  • the angiogenesis is a step in which new capillaries are formed by reconstitution of blood vessels by the decomposition of the vascular basement membrane by proteolytic enzymes, migration of vascular endothelial cells forming the vascular wall, proliferation, and formation of tubes (vascules) by vascular endothelial cell differentiation. It takes place through a series of sequential steps, including.
  • angiogenesis is strictly regulated by various negative and positive regulators (Folkman and Cotran., Int. Rev. Exp. Patho., 16, 207-248, 1976). If uncontrolled, it causes pathological disorders such as diabetic retinopathy, rheumatoid arthritis, inflammation, endometriosis, decreased vision due to aging, psoriasis and hemangioma.
  • the diseases that are related to angiogenesis can be broadly classified into inflammatory diseases such as arthritis, ophthalmic diseases such as diabetic retinopathy, dermatological diseases such as psoriasis and cancer.
  • Ocular diseases caused by angiogenesis include macular degeneration, diabetic retinopathy in which the capillaries in the retina invade the vitreous body due to complications of diabetes mellitus, retinopathy in premature infants, and neovascular glaucoma. There are diseases such as these, and these diseases cause millions of people worldwide to become blind every year.
  • arthritis is caused by autoimmune abnormalities, but chronic inflammation caused by the synovial cavity in the course of the disease is known to induce angiogenesis
  • new capillaries are diseases caused by cartilage destruction by invading joints.
  • psoriasis is a chronic proliferative disease that occurs on the skin, in order to rapidly proliferate a lot of blood must be supplied because angiogenesis is bound to actively occur. Therefore, the discovery of substances that inhibit angiogenesis is widely used in the treatment of diseases such as diabetic retinopathy, rheumatoid arthritis, inflammation, endometriosis, decreased vision due to aging, psoriasis, hemangioma, etc. As a result, academia and industry are continuing to research and develop new materials having these functions.
  • abnormal angiogenesis plays a role in supplying the nutrition and oxygen necessary for tumor growth and proliferation, and the neocapillaries that penetrate the tumor provide an opportunity for metastatic cancer cells to enter the blood circulation, which can cause cancer cells to metastasize.
  • angiogenesis inhibitors include a mechanism for reducing the activity of specific angiogenesis promoters, a mechanism for inhibiting the growth or death of vascular endothelial cells, and angiogenesis. It can be classified into four types, which play a role in inhibiting the action of promoters or indirect factors regulating endothelial cell survival factors and increasing the activity of angiogenesis inhibitors present in the body.
  • angiostatin Angiogenesis inhibitors such as endostatin, PK5, prothrombin kringle 2 are well known (O'Relly, MS et al. Cell., 79, 315-328, 1994; Lee. TH, Biol.
  • neo-N-methyl acid salbamide (NMSSV) of the cyclic pentadepsipeptide of the present invention is composed of four elements with sansalvaamide A (Sansalvamide: San A) and N-methyl acid salvamide (N-Methylsansalvamide). Cyclic pentapsipeptides with different binding order of amino acids and one hydroxy acid, which have not been reported in the organosynthesized homologues using acid salbamide A, N-methyl acid salbamide and these as basic ring structures There is nothing known regarding the inhibitory activity of angiogenesis.
  • NMSSV neo-N-methyl acid salbamide
  • an object of the present invention is to provide a pharmaceutical composition for inhibiting angiogenesis (Angiogenesis) comprising a cyclic pentapsipeptide of the formula (1) as an active ingredient.
  • the present invention provides a pharmaceutical composition for inhibiting angiogenesis (Angiogenesis) comprising a cyclic pentapsipeptide of the formula (1) as an active ingredient.
  • the composition may prevent or treat diseases caused by angiogenesis.
  • the disease is arthritis, rheumatoid arthritis, chronic inflammation, osteoarthritis, diabetic retionpathy, diabetic retinopathy, neovascular glaucoma, corneal disease caused by neovascularization, Macular degeneration / macular degeneration, spot degeneration, pterygium, retinal degeneration, erythematosis, proliferative retinopathy, posterior capsular fibrosis, granuloconjunctivitis, psoriasis, capillary dilatation, purulent granulomas, psoriasis, seborrheic dermatitis or acne Can be.
  • the cyclic pentapsipeptide of Formula 1 may inhibit the proliferation of vascular endothelial cells.
  • the cyclic pentapsipeptide of Formula 1 is ERK1 / 2 (Extracellular signal-regulated kinases 1/2), AKT (Protein Kinase B) or eNOS (endothelial nitric oxide) in vascular endothelial cells synthase) can be inhibited.
  • the cyclic pentapsipeptide of Formula 1 may inhibit cell migration of vascular endothelial cells.
  • the cyclic pentapsipeptide of Formula 1 may inhibit tube formation by vascular endothelial cell differentiation.
  • the present invention provides a health functional food for preventing and improving angiogenesis-related diseases comprising the cyclic pentapsipeptide of Formula 1 as an active ingredient.
  • the present invention also provides a method of treating angiogenesis-related diseases comprising administering the pharmaceutical composition to a subject.
  • the present invention also provides the use of the cyclic pentapsipeptide of Formula 1 to produce a medicament for preventing or treating angiogenesis-related diseases.
  • the present invention relates to a composition for inhibiting angiogenesis (Angiogenesis) containing neo-N-methyl acid salbamide (NMSSV), a cyclic pentadepsipeptide as an active ingredient, specifically, the NMSSV of the present invention It has excellent activity of inhibiting related cell migration and tube formation, and inhibits angiogenesis induced by vascular endothelial growth factor in a concentration-dependent manner, and thus is an agent for treatment of various diseases in which angiogenesis is abnormally regulated such as angiogenesis and macular degeneration. It is expected to be useful.
  • Angiogenesis angiogenesis
  • NMSSV neo-N-methyl acid salbamide
  • 1 is a diagram confirming the inhibitory effect of VEGF-induced proliferation of HUVEC by NMSSV through MTT analysis.
  • FIG. 2 is a diagram confirming the phosphorylation inhibitory effect of VEGF-induced ERK1 / 2, AKT and eNOS by HUVEC by NMSSV.
  • FIG. 3 is a diagram confirming the inhibitory effect of VEGF-induced colony tube formation of HUVEC by NMSSV.
  • FIG. 4 is a diagram confirming the effect of inhibiting VEGF-induced cell migration of HUVEC by NMSSV.
  • FIG. 5 is a diagram confirming the inhibitory effect of VEGF-induced invasion of HUVEC by NMSSV.
  • FIG. 6 is a diagram confirming the ex vivo VEGF-induced angiogenesis inhibitory effect by NMSSV through aortic ring analysis.
  • FIG. 7 is a diagram confirming the effect of inhibiting in vivo VEGF-induced angiogenesis by NMSSV through Matrigel plug analysis.
  • Figure 9 confirms the decrease in vascular endothelial cell production by NMSSV administration in CNV animal model through immunofluorescence staining.
  • the present inventors induce the vascular endothelial growth factor of NMSSV based on the fact that neo-N-methyl acid salbamide (NMSSV), a cyclic pentadepsipeptide, exhibits the activity of inhibiting cell migration and tube formation associated with angiogenesis.
  • NMSSV neo-N-methyl acid salbamide
  • the present invention was completed based on the concentration-dependent inhibitory effect of the angiogenesis.
  • the present invention provides a pharmaceutical composition for inhibiting angiogenesis (Angiogenesis) comprising the cyclic pentapsipeptide of the formula (1) as an active ingredient.
  • Angiogenesis angiogenesis
  • the "cyclic pentadepsipeptide” is a compound having a structure represented by Chemical Formula 1, and may be separated from natural products or artificially synthesized, and it will be apparent to those skilled in the art having the same effect even if artificially synthesized.
  • NMSSV nucleophilicity parameter function SSV
  • KCCM10881P Fusarium solani KCCM90040 (Accession No .: KCCM10881P).
  • the pharmaceutical composition of the present invention can ameliorate, prevent or treat a disease caused by angiogenesis, and is a disease to be improved, prevented or treated by the composition of the present invention.
  • Disease "or” disease caused by angiogenesis refers to a disease caused by abnormally progressing neovascularization.
  • angiogenesis-related disease and “angiogenesis-induced disease” may be described interchangeably.
  • Angiogenesis-related diseases that can be prevented or treated by the compositions of the present invention include angiogenesis-dependent cancer, benign tumors, arthritis, rheumatoid arthritis, chronic inflammation, osteoarthritis, diabetic retionpathy, prematurity retinopathy , Neovascular glaucoma, corneal disease caused by neovascularization, macular degeneration, spot degeneration, pterygium, retinal degeneration, erythematosis, proliferative retinopathy, posterior capsular fibrosis, granular conjunctivitis, psoriasis, capillary Dilatation, purulent granulomas, psoriasis, erythematosis, seborrheic dermatitis or acne, and the like, preferably inflammatory diseases caused by abnormal angiogenesis, ophthalmic diseases, and / or dermatological diseases, more specifically, It may be macular degeneration or arthritis, but is not limited thereto.
  • the cyclic pentapsipeptide of the present invention can perform treatment or prevention of angiogenesis-related diseases through inhibition of tube formation by cell migration and vascular endothelial cell differentiation.
  • prevention means any action that inhibits or delays the onset of angiogenesis-related diseases by administration of the pharmaceutical composition according to the present invention.
  • treatment used in the present invention means any action that improves or advantageously changes the symptoms for angiogenesis-related diseases by administration of the pharmaceutical composition according to the present invention.
  • the pharmaceutical composition according to the present invention may include a pharmaceutically acceptable carrier in addition to the active ingredient.
  • the pharmaceutically acceptable carrier is commonly used in the formulation, lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia rubber, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose , Polyvinyl pyrrolidone, cellulose, water, syrup, methyl cellulose, methylhydroxybenzoate, propyl hydroxybenzoate, talc, magnesium stearate and mineral oil, and the like.
  • it may further include a lubricant, wetting agent, sweetener, flavoring agent, emulsifier, suspending agent, preservative and the like.
  • the cyclic pentapsipeptide represented by Formula 1 of the present invention may be used in the form of a pharmaceutically acceptable salt, and as the salt, an acid addition salt formed by a pharmaceutically acceptable free acid is useful.
  • Acid addition salts include inorganic acids such as hydrochloric acid, nitric acid, phosphoric acid, sulfuric acid, hydrobromic acid, hydroiodic acid, nitrous acid or phosphorous acid and aliphatic mono and dicarboxylates, phenyl-substituted alkanoates, hydroxy alkanoates and alkanes. Obtained from non-toxic organic acids such as dioates, aromatic acids, aliphatic and aromatic sulfonic acids.
  • Such pharmaceutically nontoxic salts include sulfate, pyrosulfate, bisulfate, sulfite, bisulfite, nitrate, phosphate, monohydrogen phosphate, dihydrogen phosphate, metaphosphate, pyrophosphate chloride, bromide, and iodide.
  • Acid addition salt according to the present invention is dissolved in a conventional method, for example, the cyclic pentapsipeptide represented by the formula (1) in an excess of an aqueous acid solution, the salt is a water miscible organic solvent, for example methanol, It can be prepared by precipitation with ethanol, acetone or acetonitrile. The mixture may also be prepared by evaporation of a solvent or excess acid to evaporate or by precipitation filtration of the precipitated salt.
  • a water miscible organic solvent for example methanol
  • the mixture may also be prepared by evaporation of a solvent or excess acid to evaporate or by precipitation filtration of the precipitated salt.
  • Bases can also be used to make pharmaceutically acceptable metal salts.
  • An alkali metal or alkaline earth metal salt is obtained by, for example, dissolving a compound in an excess of alkali metal hydroxide or alkaline earth metal hydroxide solution, filtering the insoluble compound salt, and evaporating and drying the filtrate. At this time, it is pharmaceutically suitable to prepare sodium, potassium or calcium salt as the metal salt.
  • corresponding silver salts are obtained by reacting an alkali metal or alkaline earth metal salt with a suitable negative salt (eg, silver nitrate).
  • compositions of the present invention can be administered orally or parenterally (eg, applied intravenously, subcutaneously, intraperitoneally or topically) according to the desired method, and the dosage is determined by the condition and weight of the patient, Depending on the extent, drug form, route of administration, and time, it may be appropriately selected by those skilled in the art.
  • the pharmaceutical composition of the present invention is administered in a pharmaceutically effective amount.
  • pharmaceutically effective amount means an amount sufficient to treat a disease at a reasonable benefit / risk ratio applicable to medical treatment, and the effective dose level is the type of disease, severity, activity of the drug, It may be determined according to the sensitivity to the drug, the time of administration, the route of administration and the rate of release, the duration of treatment, factors including the drug used concurrently and other factors well known in the medical field.
  • the pharmaceutical compositions according to the present invention may be administered as individual therapeutic agents or in combination with other therapeutic agents, may be administered sequentially or simultaneously with conventional therapeutic agents, and may be single or multiple administrations. Taking all of the above factors into consideration, it is important to administer an amount that can obtain the maximum effect in a minimum amount without side effects, which can be easily determined by those skilled in the art.
  • the effective amount of the pharmaceutical composition of the present invention may vary depending on the age, sex, condition, weight of the patient, the absorption of the active ingredient in the body, the inactivation rate and excretion rate, the type of disease, the drug used in general 100 to 500 mg / kg body weight may be administered daily or every other day, or divided into 1 to 3 times a day.
  • the dosage may be increased or decreased depending on the route of administration, the severity of obesity, sex, weight, age, etc., and the above dosage does not limit the scope of the present invention in any way.
  • the present inventors treated with NMSSV to human Umbilical Vein Endothelial cells (HUVECs), which are human vascular endothelial cells, in order to confirm the cell migration and tube formation inhibitory effects of the cyclic pentapsipeptide of the present invention.
  • UUVECs Umbilical Vein Endothelial cells
  • VEGF Vascular endothelial growth factor
  • the present inventors produced a CNV animal model induced choroidal neovascularization by irradiating a laser, and confirmed the size of the lesion and distribution of vascular endothelial cells according to NMSSV treatment, NMSSV treatment As the size and / or extent of lesions decreases, the vascular endothelial cells are distributed in a relatively narrow space, and NMSSV suppresses abnormal blood vessel formation and inhibits the leakage of fluid from blood vessels. And / or has a therapeutic effect on diseases caused by angiogenesis and / or vascular damage in the choroid (see FIGS. 8 and 9).
  • the cyclic pentadepsipeptide of the present invention has excellent activity of inhibiting cell migration and tube formation associated with angiogenesis, and inhibits angiogenesis induced by vascular endothelial growth factor in a concentration-dependent manner, It can be usefully used as an active ingredient, and in particular, since it suppresses abnormal angiogenesis and leakage from blood vessels in the retina or choroid, it can be useful in a composition for the treatment of diseases caused by angiogenesis.
  • the present invention can provide a health functional food composition for preventing or improving angiogenesis-related diseases comprising the cyclic pentapsipeptide of the formula (1) as an active ingredient.
  • the term " improvement" means any action that at least reduces the parameters associated with the condition being treated, such as the extent of symptoms.
  • the functional food composition may be used simultaneously or separately with a medicament for treatment before or after the onset of the disease in order to prevent or improve angiogenesis-related diseases.
  • the term "health functional food” refers to a biological defense of a food group or a food composition which has added value to the food by using physical, biochemical, or biotechnological techniques to act and express the function of the food for a specific purpose. It means a food processed and designed to fully express the body's regulatory functions related to rhythm control, disease prevention and recovery to the living body, which may further include food acceptable food additives, and is a suitable carrier commonly used. It may further include excipients and diluents.
  • the health functional food composition of the present invention is a variety of nutrients, vitamins, minerals (electrolytes), flavors such as synthetic flavors and natural flavors, colorants and fillers (such as cheese, chocolate), pectic acid and salts thereof, alginic acid And salts thereof, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated beverages, and the like, which components can be used independently or in combination.
  • the amount of the composition may include 0.001% to 90% by weight of the total weight of the health functional food, preferably 0.1% to 40% by weight, may be less than the above range for long-term intake use
  • the active ingredient is not limited to the above range since there is no problem in terms of safety and can be used in an amount above the above range.
  • the present invention provides a method for treating angiogenesis-related diseases comprising administering the pharmaceutical / food composition to a subject.
  • subject means a subject in need of treatment for a disease, and more specifically, a mammal, such as a primate, mouse, dog, cat, horse and cow, which is human or non-human. .
  • VEGF Human recombinant VEGF was purchased from R & D Systems (Minneapolis, Minn.) And ERK, phospho-ERK, AKT, phospho-AKT, eNOS, phospho-eNOS, and antibodies were purchased from Cell Signaling (Danvers, MA).
  • Human umbilical vein endothelial cells were cultured to 80-90% confluence, followed by an additional 6 hours in M199 medium with 1% FBS, followed by various concentrations in the presence of VEGF (50 ng / ml).
  • VEGF 50 ng / ml
  • NMSSV neo-N-methyl acid salbamide
  • the effect of neo-N-methylsulfamide (NMSSV) on the viability and growth inhibition of HUVEC cells was analyzed by MTT (3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyltetrazolium bromide) assay. Measured.
  • test substance administration group and VEGF 50ng / ml were set up, and each group consisted of 6 8-week-old C57BL / 6 mice.
  • the matrigel plug was removed and photographed, and the amount of hemoglobin was measured and purified using the Drabkin method (Drabkin reagent kit 525, Sigma-Aldrich, Louis, MO).
  • 0.6 ml of the dorsal subcutaneous of test animals was administered using a 1 ml syringe.
  • the medium is suctioned and then the bottom of the plate is constantly scraped with a yellow tip to form a line, washed with PBS and the presence of VEGF (50 ng / ml).
  • VEGF 50 ng / ml
  • Neo-N-methyl acid salbamide (NMSSV) and VEGF-containing medium was added 200 ⁇ l and observed while incubating in a 37 °C CO 2 incubator. Incubated for 14 days after the test material treatment. On the 7th and 14th days of culture, the inverted microscope (OLYMPUS, CKX41) was observed and photographed.
  • HUVECs cells were cultured to 80-90% confluence in plate coated with Matrigel (Collaborative Biomedical Products, Bedford, Mass.), followeded by an additional 6 hours in M199 medium with 1% FBS, followed by VEGF (50 ng / ml) and treated with various concentrations of neo-N-methyl acid salbamide (NMSSV) for 24 hours. Tube formation was photographed using a phase-contrast microscope.
  • a mouse model (CNV, choroidal neovascularization) was generated by inducing laser choroidal neovascularization to have lesions similar to those of Wet-AMD. More specifically, 6-week-old C57BL / 6, male mice were purchased from the orient, purified for 1 week, and then anesthetized to expand the pupil. Subsequently, a diode green laser (532 nm, 200 mW, 0.8 sec, 50 ⁇ m, photocoagulator) was irradiated around the optic nerve at 12, 3, 6, and 9 o'clock directions, causing damage to 3-4 sites per eye. Animal models were produced.
  • CNV choroidal neovascularization
  • Example 1-9-1 the mouse was injured by a laser and anesthetized after 1 day, and then 2 ⁇ L was injected into the vitreous body once by using a 34 gauge needle (Intravitreal injection). Angiography and CNV staining and imaging were then performed 14 days after laser irradiation.
  • mice After 14 days of laser irradiation, the mice were shaken and pulsarone was injected at 80 mg / kg into the abdominal cavity for general anesthesia, followed by 1% Fluorescein intraperitoneally. After imaging using a MicronIV device, the CNV lesion area was measured using Image-Pro Plus software.
  • mice were removed from the eye and fixed in 4% paraformaldehyde for 1 hour. Subsequently, the cornea and lens were cut with scissors under a microscope, and the retina was carefully removed with forceps to prepare a retinal pigment epithelium (RPE) / choroid / scleral complex tissue. Retinal flat mount composite tissue was stirred for 1 hour in PBS containing 5% bovine serum albumin and 0.5% Triton X-100, and washed with isolectin B4 (Invitrogen, USA). Diluted to 1: 100 and reacted overnight at 4 ° C.
  • RPE retinal pigment epithelium
  • the tissue was fixed with a fluorescence mounting medium (Vector, USA) on a slide glass, covered with a cover slip, and observed and photographed using a fluorescence microscope. CNV lesion area was measured using Image-Pro Plus software.
  • Neo -N- Methyl acid salamide ( NMSSV )On by HUVEC Of human umbilical vein endothelial cells VEGF (Vascular endothelial growth factor) -induced proliferation inhibitory effect
  • VEGF 50 ng / ml
  • NMSSV Peak 2, hereinafter P2
  • VEGF 50 ng / ml
  • VEGF extracellular signal-regulated kinases 1/2 (ERK1 / 2) and AKT / eNOS (AKT / endothelial nitric oxide synthase) pathways.
  • ERK1 / 2 extracellular signal-regulated kinases 1/2
  • AKT / eNOS AKT / endothelial nitric oxide synthase
  • VEGF induced the vascular network of HUVEC in Matrigel
  • VEGF-induced colony tube formation of HUVEC was greatly inhibited by the addition of P2.
  • VEGF-induced migration of HUVECs was significantly decreased during P2 treatment, and as shown in FIG. 5, invasion by HUVECs through Matrigel was blocked when P2 was present. .
  • Example 5 On NMSSV By in vitro ( ex vivo ) VEGF -Induced angiogenesis and in vivo ( in vivo ) Confirmation of angiogenesis inhibitory effect
  • VEGF-containing Matrigel was dark red and filled with blood vessels compared to the control, while P2 added to Matrigel turned pale yellow, and compared to VEGF alone It was found that angiogenesis was reduced.
  • P2 continuously inhibits the hemoglobin content of the Matrigel plug treated with VEGF.
  • Contrast agent leaks into a certain space, such as tissue or cell gap, or the pigment is loosely adhered to the tissue, resulting in abnormally reduced or lost fluorescence staining (fluorescein leakage). Therefore, the contrast agent leakage according to NMSSV administration in the animal model produced by the method of Example 1-9 was compared with the control.
  • isolectin B4 immunostaining experiment which is a vascular endothelial cell marker, was performed to confirm the effect of reducing CNV lesions according to the NMSSV administration.
  • NMSSV-administered and unadministered eyeballs were removed to remove the lens and vitreous bodies and to obtain retinal pigment epithelium-choroid-sclera tissue. It was.
  • the area stained with isolectin B4, known as a vascular endothelial cell-specific marker was measured to determine the CNV area in the control and NMSSV-administered groups. The area of all CNV lesions in each group was measured and averaged by group to determine whether there was a difference in CNV area.
  • NMSSV a peptide isolated from potato
  • the present invention relates to a composition for inhibiting angiogenesis (Angiogenesis) containing neo-N-methyl acid salbamide (NMSSV), a cyclic pentadepsipeptide as an active ingredient, specifically, the NMSSV of the present invention
  • NMSSV neo-N-methyl acid salbamide
  • the composition according to the present invention is expected to be used in various aspects, such as for medical use, quasi-drug, functional food.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Chemical & Material Sciences (AREA)
  • Veterinary Medicine (AREA)
  • Public Health (AREA)
  • Medicinal Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Immunology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • General Chemical & Material Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Epidemiology (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Organic Chemistry (AREA)
  • Dermatology (AREA)
  • Ophthalmology & Optometry (AREA)
  • Orthopedic Medicine & Surgery (AREA)
  • Rheumatology (AREA)
  • Physical Education & Sports Medicine (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
  • Peptides Or Proteins (AREA)

Abstract

La présente invention concerne une composition pour l'inhibition de l'angiogenèse comprenant du néo-N-méthylsansalvamide (NMSSV), qui est un penta-depsipeptide cyclique, en tant que principe actif. Spécifiquement, étant donné que le NMSSV de la présente invention présente une excellente activité en matière d'inhibition de la migration cellulaire et de la formation de tubes associées à l'angiogenèse et qu'il inhibe l'angiogenèse induite par un facteur de croissance de l'endothélium vasculaire d'une manière dépendant de la concentration, on s'attend à ce que la présente invention puisse être utilisée avec profit pour l'inhibition de l'angiogenèse et en tant qu'agent thérapeutique pour diverses maladies dans lesquelles on observe des anomalies au niveau de la régulation de l'angiogenèse.
PCT/KR2018/001560 2017-02-09 2018-02-06 Composition pharmaceutique pour inhibition de l'angiogenèse contenant un penta-depsipeptide cyclique en tant que principe actif Ceased WO2018147615A1 (fr)

Priority Applications (2)

Application Number Priority Date Filing Date Title
US16/320,502 US10709757B2 (en) 2017-02-09 2018-02-06 Pharmaceutical composition for anti-angiogenesis containing cyclic pentadepsipeptide as an effective ingredient
CN201880002835.7A CN109689083B (zh) 2017-02-09 2018-02-06 含作为有效成分的环五缩酚酸肽的抗血管生成药物组合物

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
KR10-2017-0018103 2017-02-09
KR20170018103 2017-02-09
KR1020180011886A KR102022631B1 (ko) 2017-02-09 2018-01-31 고리형 펜타뎁시펩타이드를 유효성분으로 함유하는 혈관신생 억제용 약학적 조성물
KR10-2018-0011886 2018-01-31

Publications (1)

Publication Number Publication Date
WO2018147615A1 true WO2018147615A1 (fr) 2018-08-16

Family

ID=62948404

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/KR2018/001560 Ceased WO2018147615A1 (fr) 2017-02-09 2018-02-06 Composition pharmaceutique pour inhibition de l'angiogenèse contenant un penta-depsipeptide cyclique en tant que principe actif

Country Status (2)

Country Link
FR (1) FR3062570B1 (fr)
WO (1) WO2018147615A1 (fr)

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20110076845A (ko) * 2009-12-29 2011-07-06 이화여자대학교 산학협력단 혈관신생 억제용 약제학적 조성물
KR101119561B1 (ko) * 2009-01-02 2012-03-07 중앙대학교 산학협력단 고리형 펜타뎁시펩타이드를 생산하는 푸사리움속 미생물
KR101120565B1 (ko) * 2009-01-02 2012-03-09 중앙대학교 산학협력단 신규 고리형 펜타뎁시펩타이드(ⅰ) 및 그 이용
EP2293846B1 (fr) * 2008-05-22 2013-07-10 Karus Therapeutics Limited Depsipeptides et leur utilisation thérapeutique

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP2293846B1 (fr) * 2008-05-22 2013-07-10 Karus Therapeutics Limited Depsipeptides et leur utilisation thérapeutique
KR101119561B1 (ko) * 2009-01-02 2012-03-07 중앙대학교 산학협력단 고리형 펜타뎁시펩타이드를 생산하는 푸사리움속 미생물
KR101120565B1 (ko) * 2009-01-02 2012-03-09 중앙대학교 산학협력단 신규 고리형 펜타뎁시펩타이드(ⅰ) 및 그 이용
KR20110076845A (ko) * 2009-12-29 2011-07-06 이화여자대학교 산학협력단 혈관신생 억제용 약제학적 조성물

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
LEE, H.-S. ET AL.: "Optimization of Culture Conditions of Fusarium Solani for the Production of neoN-methylsansalvamide", BIOSCI. BIOTECHNOL. BIOCHEM., vol. 78, no. 8, 12 June 2014 (2014-06-12), pages 1421 - 1427, XP055537160 *
SONG, H.-H. ET AL.: "A New Cytotoxic Cyclic Pentadepsipeptide, Neo-N-methylsansalvamide Produced by Fusarium Solani KCCM90040, Isolated from Potato", FOOD CHEMISTRY, vol. 126, no. 2, 2011, pages 472 - 478, XP027571811 *

Also Published As

Publication number Publication date
FR3062570B1 (fr) 2021-03-12
FR3062570A1 (fr) 2018-08-10

Similar Documents

Publication Publication Date Title
RU2470635C2 (ru) Профилактическое или терапевтическое средство, применяемое при заболеваниях заднего отдела глаза
KR102022631B1 (ko) 고리형 펜타뎁시펩타이드를 유효성분으로 함유하는 혈관신생 억제용 약학적 조성물
TW201038544A (en) Anti-neurodegenerative diseases agents
JP2024150780A (ja) 眼組織の線維化抑制用組成物
WO2021101344A1 (fr) Composition de gouttes oculaires pour la prévention ou le traitement d'une maladie oculaire
WO2024043706A1 (fr) Nouvel agrégat moléculaire d'ingrédient pharmacologique actif et composition pharmaceutique le comprenant
JP5503879B2 (ja) 視神経障害を伴う眼疾患の予防又は治療剤
US20160213671A1 (en) Prophylaxis and treatment of a neurodegenerative disease not based on a protein-folding disorder
WO2018147615A1 (fr) Composition pharmaceutique pour inhibition de l'angiogenèse contenant un penta-depsipeptide cyclique en tant que principe actif
WO2024014792A1 (fr) Composition pharmaceutique pour traiter des maladies oculaires comprenant des exosomes dérivés d'organoïdes rétiniens
JP2006523461A (ja) 羊ヒアルロニダーゼを単離し精製する方法
CN109789115A (zh) 用于治疗眼疾的方法
Ejstrup et al. Electrophysiological consequences of experimental branch retinal vein occlusion in pigs and the effect of dorzolamide
WO2020091430A1 (fr) Composition de prévention ou de traitement de la dégénérescence maculaire
KR20210135720A (ko) 황반변성의 완화, 억제, 치료 또는 예방용 조성물
KR100854058B1 (ko) 스테로이드를 유효 성분으로 하는 망맥락막 질환 치료제
JP6708630B2 (ja) 角膜障害の予防又は治療剤
US20210061800A1 (en) Compounds for treating eye diseases and methods thereof
CN113768929B (zh) Fdi化合物在眼科疾病中的用途
WO2022240165A1 (fr) Composition pharmaceutique pour la prévention ou le traitement de la dégénérescence maculaire liée à l'âge
WO2012115439A2 (fr) Composition comprenant des régulateurs d'expression ou d'activité d'akap12 pour traiter les maladies du système nerveux central
EP0719556A2 (fr) Agent pour le traitement de la kératopathie
WO2011046239A1 (fr) Nouvelle utilisation d'un composé imidazole
WO2023033483A1 (fr) Composition pharmaceutique de prévention ou de traitement d'une maladie oculaire diabétique comprenant un inhibiteur de sglt-2
WO2023014117A1 (fr) Composition de gouttes oculaires pour le traitement du syndrome de l'oeil sec contenant une nouvelle recoflavone et son procédé de préparation

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: 18751178

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: 18751178

Country of ref document: EP

Kind code of ref document: A1