LU504076B1 - Application of compound in preparation of hdac2 gene activator - Google Patents

Application of compound in preparation of hdac2 gene activator Download PDF

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Publication number
LU504076B1
LU504076B1 LU504076A LU504076A LU504076B1 LU 504076 B1 LU504076 B1 LU 504076B1 LU 504076 A LU504076 A LU 504076A LU 504076 A LU504076 A LU 504076A LU 504076 B1 LU504076 B1 LU 504076B1
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Luxembourg
Prior art keywords
hdac2
gene activator
methyl
compound
dihydropyrazol
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Application number
LU504076A
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German (de)
Inventor
Qian Wu
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The Second Affiliated Hospital And Yuying Childrens Hospital Of Wenzhou Medical Univ
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    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00Medicinal preparations containing organic active ingredients
    • A61K31/33Heterocyclic compounds
    • A61K31/395Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/41Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
    • A61K31/4151,2-Diazoles
    • A61K31/41521,2-Diazoles having oxo groups directly attached to the heterocyclic ring, e.g. antipyrine, phenylbutazone, sulfinpyrazone
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00Medicinal preparations characterised by special physical form
    • A61K9/0012Galenical forms characterised by the site of application
    • A61K9/0019Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61PSPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P43/00Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
    • YGENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02PCLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
    • Y02P20/00Technologies relating to chemical industry
    • Y02P20/50Improvements relating to the production of bulk chemicals
    • Y02P20/55Design of synthesis routes, e.g. reducing the use of auxiliary or protecting groups

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  • Health & Medical Sciences (AREA)
  • Veterinary Medicine (AREA)
  • Chemical & Material Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Epidemiology (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Organic Chemistry (AREA)
  • Dermatology (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Abstract

Disclosed is the application of a compound 2-(3-aminophenyl)-5-methyl-2,4-dihydropyrazol-3-one hydrochloride in the preparation of an HDAC2 gene activator. The present invention has revealed that the compound 2-(3-aminophenyl)-5-methyl-2,4-dihydropyrazol-3-one hydrochloride is capable of up-regulating the HDAC2 gene expression, playing the role of HDAC2 gene activator, thereby providing a new option for the treatment of HDAC2-associated diseases.

Description

BL-5641
LU504076
APPLICATION OF COMPOUND IN PREPARATION OF HDAC2 GENE
ACTIVATOR
TECHNICAL FIELD
[01] The present invention relates to the technical field of gene activators, and particularly relates to the application of a compound in the preparation of an HDAC2 gene activator.
BACKGROUND ART
[02] Histone deacetylase (HAT) and histone deacetylase (HDAC) are a pair of functional antagonistic proteases that regulate the acetylation state of histones, and jointly regulate the normal physiological functions of human body. Misexpression or dysfunction, if any, may even lead to the occurrence of tumors.
[03] Mammalian cells express 18 HDAC enzymes, which are divided into two classes:
Class II NAD+-dependent HDACs or sirtuins (sirtuins 1 - 7) and typical zinc-dependent
HDACSs, including: Class I (HDACs 1, 2, 3 and 8), Class II (HDACs 4, 5, 7 and 9), Class IIb (HDACs 6 and 10) and Class IV (HDACI11), have become therapeutic targets a variety of diseases, and have very important scientific research value and clinical application prospects.
[04] The 2H-pyran and 4H-pyran ring heterocyclic backbone is a structural parent nucleus with a wide range of significant biological activities identified in natural compounds, such as anti-neurodegenerative diseases, anti-cancer, cytotoxicity, anti-HIV, anti-malaria, anti-bacteria, anti-hyperglycemia and anti-dyslipidemia, eg, Alzheimer's Disease,
Parkinson's Disease and Huntington's Disease. Among them, 1-phenyl-3-methyl-5-pyrazolone is used to synthesize pyrazolone antipyretic and analgesic drugs such as metamizole sodium, antipyrine and aminopyrine. However, neither report nor application of 2-(3-aminophenyl)-5-methyl-2,4-dihydropyrazol-3-one hydrochloride directly used in HDAC2 activators is available yet.
SUMMARY
[05] The present invention provides for the first time the compound 2-(3-aminophenyl)-5-methyl-2,4-dihydropyrazol-3-one hydrochloride as an active ingredient in an HDAC2 gene activator, providing a new option for the treatment of HDAC2-associated diseases.
[06] The technical scheme of the present invention: application of a compound in the preparation of an HDAC2 gene activator, where the compound is 2-(3-aminophenyl)-5-methyl-2,4-dihydropyrazol-3-one hydrochloride, and the compound structure is as follows: 1
BL-5641
Co LU504076
ON
Mo
[07] Further, the dosage form of the HDAC2 gene activator is a solid formulation or a liquid formulation.
[08] Further, the HDAC2 gene activator is administered orally or by injection.
[09] Further, the HDAC2 gene activator also contains a pharmaceutically acceptable carrier or excipient.
[10] Further, the excipient includes one or some of a solvent, a binder, a filler, a stabilizer and an emulsifier.
[11] An HDAC2 gene activator, including 2-(3-aminophenyl)-5-methyl-2,4-dihydropyrazol-3-one hydrochloride.
[12] The mass concentration of the 2-(3-aminophenyl)-5-methyl-2,4-dihydropyrazol-3-one hydrochloride in the above HDAC2 gene activator is 10 - 300 ng/mL.
[13] Compared with the prior art, the present invention provides for the first time the application of a compound 2-(3-aminophenyl)-5-methyl-2,4-dihydropyrazol-3-one hydrochloride in the preparation of an HDAC2 gene activator. The present invention has revealed that the compound 2-(3-aminophenyl)-5-methyl-2,4-dihydropyrazol-3-one hydrochloride is capable of up-regulating the HDAC2 gene expression, playing the role of
HDAC2 gene activator, thereby providing a new option for the treatment of
HDAC2-associated diseases.
BRIEF DESCRIPTION OF THE DRAWINGS
[14] FIG 1 is a schematic diagram of the structural formulae of three separate compounds numbered 50366, 01206 and 07509, respectively.
[15] FIG 2 shows the effect of three separate compounds on the expression of deacetylated proteins HDAC2 and HDACS3 in A375 cells.
[16] FIG 3 is a statistical diagram of the expression of deacetylated proteins HDAC2 and
HDAC3.
[17] FIG 4 shows the MRNA expression of HDAC in HUVEC cells after medication.
[18] FIG 5 shows the effect of the expression of proteins extracted from the skin at the wound site after external application.
[19] FIG 6 is a statistical diagram of the expression of HDAC1, HDAC2 and HDAC3 in the three groups of experiments.
[20] FIG 7 shows a comparison of the expression of deacetylated proteins HDAC2 and 2
BL-5641 . . Ce. LU504076
HDAC3 extracted from the skin at the wound site between the injection treatment group and the challenge group.
[21] FIG 8 is a statistical diagram of the expression of deacetylated proteins HDAC2 and
HDAC3 extracted from the skin at the wound site between the control group and the challenge group.
DETAILED DESCRIPTION OF THE EMBODIMENTS
[22] The present invention will be further elaborated hereafter in conjunction with accompanying drawings and the embodiments, which, however, are not taken as a basis for limiting the present invention.
[23] Example: the application of a compound in the preparation of an HDAC2 gene activator, where the compound is 2-(3-aminophenyl)-5-methyl-2,4-dihydropyrazol-3-one hydrochloride, and the compound structure is as follows:
MR pti
[24] Further, the dosage form of the HDAC2 gene activator is a solid formulation or a liquid formulation. The HDAC2 gene activator is administered orally or by injection. The
HDAC?2 gene activator also contains a pharmaceutically acceptable carrier or excipient. The excipient includes one or some of a solvent, a binder, a filler, a stabilizer and an emulsifier.
[25] Example 2: an HDAC2 gene activator, the HDAC2 gene activator includes 2-(3-aminophenyl)-5-methyl-2,4-dihydropyrazol-3-one hydrochloride. Its dosage form is a solid formulation or a liquid formulation, it is administered orally or by injection, and it further contains a pharmaceutically acceptable carrier or excipient. The excipient includes one or some of a solvent, a binder, a filler, a stabilizer and an emulsifier.
[26] Example 3: effect of 2-(3-aminophenyl)-5-methyl-2,4-dihydropyrazol-3-one hydrochloride on A375 human melanoma cells.
[27] Three separate compounds were added to A375 human melanoma cells, the proteins were then extracted, and the inhibitory effect on acetylated proteins was verified by western blotting technique. The three separate compounds are numbered as 50366, 01206 and 07509, respectively, among which Compound No. 50366 is 2-(3-aminophenyl)-5-methyl-2,4-dihydropyrazol-3-one hydrochloride, Compound No. 01206 is 1-(4-chlorophenyl )-3-methyl-2-pyrazoline-5-one, and Compound No. 07509 is 2-(4-fluorophenyl)-5-methyl-4H-pyrazol-3-one, with the structural formulae shown in FIG. 1. FIG 2 shows the effect of three separate compounds on the expression of deacetylated proteins in A375 cells. FIG 3 is a statistical diagram of the expression of each protein. The experimental results from FIG 2 and FIG 3 show that Compound No. 01206 and 07509 have 3
BL-5641 obvious inhibitory effect on acetylation, while Compound No. 50366 (namely, LUS04076 2-(3-aminophenyl)-5-methyl-2,4-dihydropyrazol-3-one hydrochloride provided by the present invention) can promote acetylation.
[28] Example 4: effect of 2-(3-aminophenyl)-5-methyl-2,4-dihydropyrazol-3-one hydrochloride on stimulating HDAC in HUVEC.
[29] HUVEC cells refer to human umbilical vein endothelial cells in the human body, and an RT-PCR assay was used to detect whether HDAC2 gene activator containing 2-(3-aminophenyl)-5-methyl-2,4-dihydropyrazol-3-one hydrochloride can promotes the increase of HDAC2 expression in HUVEC cells. FIG 4 shows the mRNA expression of
HDAC in HUVEC cells after medication. The results from FIG 2 show that, compared with the control group, the expression of HDAC2 and HDACS in the cells of the 50 ng/mL administration group was significantly increased.
[30] Example 5: effect of external application of 2-(3-aminophenyl)-5-methyl-2,4-dihydropyrazol-3-one hydrochloride on the expression of
HDAC protein at the wound site.
[31] After SD rats were shaved and disinfected, and two full-thickness wounds with a diameter of 20 mm were made on both sides of the back of the rats. The control group was treated by covering the wounds with a sterile transparent dressing for the first three days, the vaseline group was treated by covering the wounds with a sterile transparent dressing for the first three days and by reapplying the same once every two days after being covered with the vaseline, while the challenge group was treated by covering the wounds with a dressing containing 100 nmol/g of drug (containing 2-(3-aminophenyl)-5-methyl-2,4-dihydropyrazol-3-one hydrochloride) once every two days, and the wound healing was observed and recorded for 21 days. After anesthesia, the rats were killed, the proteins were extracted from the skin at the wound site, and the expression of proteins was measured by the western blotting. FIG. 5 shows the effect of the expression of proteins extracted from the skin at the wound site; and FIG 6 is a statistical diagram of the expression of HDACI, HDAC2 and HDAC3 in the three groups of experiments. The experimental results from FIG 5 and FIG 6 show that HDAC 2/3 in the challenge group were up-regulated, indicating that external application of 2-(3-aminophenyl)-5-methyl-2,4-dihydropyrazol-3-one hydrochloride could have a stimulating effect on the activation of HDAC2 level at the wound site.
[32] Example 6: effect of the injection of 2-(3-aminophenyl)-5-methyl-2,4-dihydropyrazol-3-one hydrochloride on the expression of
HDAC proteins at the wound site.
[33] After SD rats were shaved and disinfected, and two full-thickness wounds with a diameter of 20mm were made on both sides of the back of the rats. The control group was treated by covering the wounds with a sterile transparent dressing for the first three days, the vaseline group was treated by covering the wounds with a sterile transparent dressing for the first three days and by reapplying the same once every two days after being covered with the vaseline, while the challenge group was treated by injecting the normal saline containing 100 nmol/g of drug (containing 2-(3-aminophenyl)-5-methyl-2,4-dihydropyrazol-3-one hydrochloride) once every two days, and the control group was treated by injecting the normal saline. After anesthesia, the rats were killed, the proteins were extracted from the skin at the wound site, and the expression of proteins was measured by the western blotting.
The experimental results from FIG 7 and FIG 8 show that HDAC 2/3 in the challenge group 4
BL-5641 were up-regulated, indicating that injection of LUS04076 2-(3-aminophenyl)-5-methyl-2,4-dihydropyrazol-3-one hydrochloride could have a stimulating effect on the activation of HDAC?2 level at the wound site.
[34] To sum up, the present invention provides for the first time the application of a compound 2-(3-aminophenyl)-5-methyl-2,4-dihydropyrazol-3-one hydrochloride in the preparation of an HDAC2 gene activator. The present invention has revealed that the compound 2-(3-aminophenyl)-5-methyl-2,4-dihydropyrazol-3-one hydrochloride is capable of up-regulating the HDAC2 gene expression, playing the role of HDAC2 gene activator, thereby providing a new option for the treatment of HDAC2-associated diseases.

Claims (7)

BL-5641 CLAIMS LU504076
1. Application of a compound in the preparation of an HDAC2 gene activator, wherein the compound is 2-(3-aminophenyl)-5-methyl-2,4-dihydropyrazol-3-one hydrochloride.
2. The application according to claim 1, wherein the dosage form of the HDAC2 gene activator is a solid formulation or a liquid formulation.
3. The application according to claim 1, wherein the HDAC2 gene activator 1s administered orally or by injection.
4. The application according to claim 1 or claim 2, wherein the HDAC2 gene activator also contains a pharmaceutically acceptable carrier or excipient.
5. The application according to claim 4, wherein the excipient comprises one or some of a solvent, a binder, a filler, a stabilizer and an emulsifier.
6. An HDAC2 gene activator, wherein the HDAC2 gene activator comprises 2-(3-aminophenyl)-5-methyl-2,4-dihydropyrazol-3-one hydrochloride.
7. An HDAC2 gene activator according to claim 6, wherein the mass concentration of 2-(3-aminophenyl)-5-methyl-2,4-dihydropyrazol-3-one hydrochloride in the HDAC2 gene activator is 10 - 300 ng/mL. 6
LU504076A 2023-02-27 2023-04-26 Application of compound in preparation of hdac2 gene activator LU504076B1 (en)

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CN116531369A (en) * 2023-02-24 2023-08-04 杜娟娟 Application of a compound in promoting wound healing medicine

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* Cited by examiner, † Cited by third party
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JP2004115508A (en) * 2002-09-06 2004-04-15 Mitsubishi Pharma Corp Drugs for improving the function of burned skin tissue
US20120022129A1 (en) * 2008-08-18 2012-01-26 Irfan Rahman Targeting of Histone Deacetylase 2, Protein Kinase CK2, and Nuclear Factor NRF2 For Treatment of Inflammatory Diseases
US20160199463A1 (en) * 2014-12-15 2016-07-14 The Johns Hopkins University Hdac2 defends vascular endothelium from injury
CN111346095B (en) * 2020-03-14 2021-06-08 温州医科大学附属第二医院、温州医科大学附属育英儿童医院 Pharmaceutical preparations for the treatment of post-neurosurgical headaches
CN111840261A (en) * 2020-06-22 2020-10-30 广州中医药大学(广州中医药研究院) A kind of agonist of deacetylase and its application
CN116531369A (en) * 2023-02-24 2023-08-04 杜娟娟 Application of a compound in promoting wound healing medicine

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