Home Immunology & Inflammation SPHK inhibitor Opaganib (ABC294640)

연구용

Opaganib (ABC294640) SphK2 Inhibitor

제품 번호: S7174

Opaganib (ABC294640) is an orally bioavailable and selective sphingosine kinase-2 (SphK2) inhibitor with IC50 of approximately 60 μM. Phase 1/2.
Opaganib (ABC294640) SPHK inhibitor Chemical Structure

화학 구조

분자량: 380.91

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품질 관리 (Quality Control)

배치: 순도: 99.73%
99.73

세포 배양, 처리 및 작업 농도
(Cell Culture, Treatment & Working Concentration)

세포주 분석 유형 농도 배양 시간 제형 활성 설명 PMID
Sf9 Function assay Inhibition of human recombinant SphK2 expressed in Sf9 cells assessed as radiolabeled products using 10 uM sphingosine and 10 uM gamma[32P]ATP by liquid scintillation counting, Ki=9.8μM 25643074
Sf9 Function assay 2 hrs Inhibition of recombinant human full-length N-terminal His-tagged SK2 expressed in baculovirus infected Sf9 insect cells using NBD-Sph as substrate measured after 2 hrs by fluorescence based HPLC analysis, Ki=9.8μM 30889352
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화학 정보, 보관 및 안정성 (Chemical Information, Storage & Stability)

분자량 380.91 화학식

C23H25ClN2O

 보관 (수령일로부터)
CAS 번호 915385-81-8 SDF 다운로드 원액 보관

동의어 N/A Smiles C1C2CC3(CC1CC(C2)(C3)C(=O)NCC4=CC=NC=C4)C5=CC=C(C=C5)Cl

용해도 (Solubility)

In vitro
배치:

DMSO : 76 mg/mL (199.52 mM)
(수분으로 오염된 DMSO는 용해도를 감소시킬 수 있습니다. 신선하고 무수 DMSO를 사용하십시오.)

Ethanol : 28 mg/mL

Water : Insoluble

몰농도 계산기

질량 농도 부피 분자량
희석 계산기 분자량 계산기

In vivo
배치:

생체 내 제형 계산기 (투명한 용액)

1단계: 아래 정보 입력 (권장: 실험 중 손실을 고려하여 추가 동물 포함)

mg/kg g μL

2단계: 생체 내 제형 입력 (이것은 계산기일 뿐 제형이 아닙니다. 용해도 섹션에 생체 내 제형이 없는 경우 먼저 당사에 문의하십시오.)

% DMSO % % Tween 80 % ddH2O
%DMSO %

계산 결과:

작업 농도: mg/ml;

DMSO 원액 준비 방법: mg 약물 사전 용해 μL DMSO ( 원액 농도 mg/mL, 농도가 해당 약물 배치의 DMSO 용해도를 초과하는 경우 먼저 당사에 문의하십시오. )

생체 내 제형 준비 방법: 취하다 μL DMSO 원액, 다음 추가μL PEG300, 혼합하고 투명하게 한 다음 추가μL Tween 80, 혼합하고 투명하게 한 다음 추가 μL ddH2O, 혼합하고 투명하게 합니다.

생체 내 제형 준비 방법: 취하다 μL DMSO 원액, 다음 추가 μL 옥수수 기름, 혼합하고 투명하게 합니다.

참고: 1. 다음 용매를 추가하기 전에 액체가 투명한지 확인하십시오.
2. 용매를 순서대로 추가해야 합니다. 다음 용매를 추가하기 전에 이전 추가에서 얻은 용액이 투명한 용액인지 확인해야 합니다. 와동, 초음파 또는 뜨거운 물 중탕과 같은 물리적 방법을 사용하여 용해를 도울 수 있습니다.

작용 메커니즘 (Mechanism of Action)

Targets/IC50/Ki
SphK2
(Cell-free assay)
60 μM
시험관 내(In vitro)
Opaganib (ABC294640) markedly alters the ratio of ceramide/S1P consistent with inhibition of SK activity in MDA-MB-231 cells. It inhibits tumor cell proliferation with IC50 values ranging from approximately 6 to 48 μM, and impairs tumor cell migration concomitant with loss of microfilaments. This compound induces nonapoptotic cell death, morphological changes in lysosomes, formation of autophagosomes, and increases in acidic vesicles in A-498, PC-3, and MDA-MB-231 cells. In both MCF-7 and ER-transfected HEK293 cells, it decreases E2-stimulated ERE-luciferase activity.
키나아제 분석
Sphingosine Kinase Assays
The IC50 values for Opaganib (ABC294640) and DMS are determined by a newly developed HPLC-based SK activity assay. In brief, the test compounds are incubated with recombinant SK1 or SK2 and NBD-Sph in the isozyme-selective assay buffers detailed below with 1 mg/ml fatty acid-free bovine serum albumin, 100 μM ATP, and 400 μM MgCl2. The product, i.e., NBD-S1P, is separated from NBD-Sph by HPLC as follows: Waters 2795 HPLC system with a Waters 2495 fluorescence detector, C8 Chromolith RP-8e column (100 × 4.6 mm), 1 ml/min mobile phase (acetonitrile/20 mM sodium phosphate buffer, pH2.5, at 45:55). Fluorescence is monitored with excitation at 465 nm and emission at 531 nm. The ratio of NBD-S1P/(NBD-Sph + NBD-S1P) is used as a measure of SK activity. SK-isozyme selective assay buffers each contained 20 mM Tris, pH7.4, 5 mM EDTA, 5 mM EGTA, 3 mM β-mercaptoethanol, 5% glycerol, 1× protease inhibitors and 1× phosphatase inhibitors. For the SK1 assay buffer, 0.25% (final) Triton X-100 is added; and for the SK2 buffer, 1 M (final) KCl is added. Assays are run for 2 h at room temperature, and then a 1.5 volume of methanol is added to terminate the kinase reaction. After 10 min, the samples are centrifuged at 20,000g to pellet the precipitated protein, and the supernatants are analyzed by HPLC. In experiments to determine the Ki for inhibition of SK2 by this compound, the ADP Quest assay system is used to measure kinase activity in the presence of varying concentrations of sphingosine and it. To determine the effects of Opaganib on cellular SK activity, near-confluent MDA-MB-231 cells are serum-starved overnight, and then treated with varying concentrations of it. The cells are then incubated with [3H]sphingosine at a final concentration of 1 μM. The cells take up the exogenous sphingosine, which is converted to S1P via SK activity, and [3H]S1P is separated from [3H]sphingosine by extraction and quantified by scintillation counting.
생체 내(In vivo)
Opaganib (ABC294640) significantly reduces tumor growth in mice bearing mammary adenocarcinoma xenografts, associated with depletion of S1P levels. In severe combined immunodeficient mice bearing A-498 xenografts, it delays tumor growth and elevates autophagy markers. This compound also protects against liver transplantation-induced inflammation and cross-talk between innate and adaptive immunities, major events precipitating and exacerbating graft injury, and improves liver function and survival.
참조
  • [4] https://pubmed.ncbi.nlm.nih.gov/22848628/

적용 분야 (Applications)

방법 바이오마커 이미지 PMID
Western blot c-Myc pRb / Rb H3K9ac / p21 p-STAT3 / STAT3
S7174-WB1
27517489
Growth inhibition assay Cell proliferation
S7174-viability1
25122609

임상시험 정보 (Clinical Trial Information)

(데이터 출처 https://clinicaltrials.gov, 업데이트 날짜 2024-05-22)

NCT 번호 모집 조건 스폰서/협력자 시작일 단계
NCT02939807 Withdrawn
Carcinoma Hepatocellular
Medical University of South Carolina|National Cancer Institute (NCI)|Apogee Biotechnology Corporation
 September 30 2019 Phase 2
NCT03377179 Completed
Cholangiocarcinoma|Cholangiocarcinoma Non-resectable|Cholangiocarcinoma Perihilar|Cholangiocarcinoma Extrahepatic|Cholangiocarcinoma Intrahepatic
RedHill Biopharma Limited
 March 7 2018 Phase 2
NCT02757326 Terminated
Multiple Myeloma
RedHill Biopharma Limited|Apogee Biotechnology Corporation|Duke University|National Cancer Institute (NCI)
 December 13 2016 Phase 1|Phase 2
NCT02229981 Withdrawn
Diffuse Large B Cell Lymphoma|Kaposi Sarcoma
RedHill Biopharma Limited|Louisiana State University Health Sciences Center in New Orleans|Apogee Biotechnology Corporation
 July 2014 Phase 1|Phase 2
NCT01488513 Completed
Pancreatic Cancer|Unspecified Adult Solid Tumor Protocol Specific
RedHill Biopharma Limited|Apogee Biotechnology Corporation|Medical University of South Carolina|FDA Office of Orphan Products Development|National Cancer Institute (NCI)
 August 2011 Phase 1