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
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P35/00—Antineoplastic agents

Definitions

• the subject of the invention is N- ⁇ 2- [ 4- ( 4-methoxyphenyl ) -1 , 3- thiazol-2-yl ] ethyl ⁇ -2-oxo-2, 5, 6 , 7-tetrahydro-lH- cyclopenta [b] pyridine-3-carboxamide as an anti-cancer agent or efflux pump inhibitor.
• the invention finds application in medicine .
• An active efflux is a mechanism responsible for the extrusion of toxic substances and antibiotics from within cells to the external environment.
• the said mechanism is important in medicine, as it may contribute to bacterial immunity to antibiotics or chemotherapy during cancer
• ABC type efflux pumps are known in the form of protein transporters located in the cellular membranes of all cell types. As active transporters, they require a source of chemical energy, which may be provided by, amongst others, adenosine triphosphate hydrolysis
• the second subject of the invention is N- ⁇ 2-[4-(4- 30 methoxyphenyl ) -1, 3-thiazol-2-yl] ethyl ⁇ -2-oxo-2, 5, 6, 7- tetrahydro-l/i-cyclopenta [b] pyridine-3-carboxamide to be used as an efflux pump inhibitor, preferably type ABC class Bl, B8, CI and G2.
• the compound obtained in example 1 was dissolved in 100% DMSO, which in the test constituted solvent control whereas cisplatin was the reference compound.
• Starting solutions (stock solutions) of the tested substance from example 1 with a concentration of 10 mM were prepared for each experiment ex tempore, dissolving 0.8 ⁇ of the compound in 400 ⁇ test medium OR ( OptiMEM+RPMI , 5% FBS, antibiotics ( SP ( streptomycin, penicillin) ) .
• solutions of the tested substances were prepared together with a solvent control, obtaining final compound concentrations of 10, 1, 0.1, 0.01 ⁇ and 0.1, 0.01, 0.001, 0.0001 % DMSO.
• Cytotoxic activity was determined as follows. Approximately 10,000 cells in 100 ⁇ dedicated culture medium were placed into a 96-well storage plate, medium without cells placed into 3 wells. After 24h incubation at 37°C in 5% CO 2 and 95 % air, solutions of tested compounds were applied (3 times for each dilution) to the last row of the 96-well plate which constituted control cells and control medium to which only OR test medium was added. Half of a 96-well storage plate was prepared in a similar manner for solvent control (DMSO) and cisplatin. After 72 h, 20 ⁇ of MTT solution was added to each well, then after 4 h of incubation 80 ⁇ of lysis buffer was added and a reading was made using spectrophotometer set at 545 nm.
• DMSO solvent control
• Cis-Pt 2.65 ⁇ 1.19 3.30 ⁇ 2.25 2.26 ⁇ 1.52 DMSO solvent control did not affect tested cells and proliferation inhibition did not exceed 15% in any used cell line.
• the addition of compound from example 1 led to significant toxicity for LoVoDX line, but not for LoVo line. 5 Toxicity was caused by doxorubicin, which the LoVoDx line may remove aided by active efflux pumps. Blocking efflux pumps by the compound from example 1 is the reason for doxorubicin toxicity for LoVoDX line. Toxicity for cisplatin is caused by the specificity of efflux pumps overexpression .
• Table 3 shows cell proliferation inhibition following DMSO [%] and Cis-Pt [ug/ml] treatment.
• Cisplatin toxicity for cell lines is caused by a lack of mechanism removing cisplatin from cells.
• BALB/3T3 line normally does not contain overexpressed ABC type efflux pumps which are able to remove cisplatin.
• LoVoDx does contain such overexpressed transporters but these may be unspecific for cisplatin .
• Table 4 shows cell proliferation inhibition relations to the used compound concentration
• Cell proliferation inhibition by the compound obtained in example 1 indicates compound toxicity.
• proliferation inhibition does not exceed 25%.
• LoVoDX line exhibits proliferation inhibition of more than 50% in the presence of doxorubicin in the same compound concentration obtained in example 1, which indicates a lack of active doxorubicin removal mechanism. The lack of mechanism is caused by inhibition of ABC type efflux pump which remove doxorubicin as normally the LoVoDX line overexpresses such pumps.

Landscapes

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

Applications Claiming Priority (2)

Publications (1)

Family

ID=52004034

Family Applications (1)

Country Status (2)

Citations (4)

• 2013
  • 2013-10-23 PL PL405743A patent/PL226024B1/pl unknown
• 2014
  • 2014-10-23 WO PCT/PL2014/050067 patent/WO2015060738A1/en active Application Filing

Patent Citations (4)

Non-Patent Citations (3)

Also Published As

Similar Documents

Legal Events