JPI-547 targets PARP1, PARP2, tankyrase 1 (TNKS1), and tankyrase 2 (TNKS2). PARP1 and PARP2 are involved in DNA repair, while tankyrases are involved in the Wnt/β-catenin signaling pathway. By inhibiting both PARP and tankyrase, JPI-547 blocks both DNA repair and Wnt signaling, which are important for cancer cell survival and proliferation. The compound's dual mechanism of action may overcome resistance to PARP inhibitor monotherapy. JPI-547's antitumor activity has been demonstrated in BRCA-deficient cancers.

In vitro, JPI-547 demonstrates potent inhibition of PARP and tankyrase. The compound's activity is assessed using biochemical assays measuring PARP and tankyrase enzymatic activity. JPI-547 also shows activity in cell-based assays, where it inhibits PARP-mediated DNA repair and tankyrase-mediated Wnt signaling, leading to cell death in cancer cells. The compound's dual mechanism of action makes it a valuable tool for studying the roles of PARP and tankyrase in cancer biology.

In vivo, JPI-547 has demonstrated antitumor activity in preclinical models. The compound is orally active and has been evaluated in xenograft models of cancer. JPI-547's dual inhibition of PARP and tankyrase may offer advantages over PARP inhibitor monotherapy in certain cancers.

JPI-547's inhibition of PARP1/2 and tankyrase1/2 is assessed using biochemical enzyme assays. Recombinant PARP1, PARP2, TNKS1, or TNKS2 is incubated with the test compound and the appropriate substrate; enzymatic activity is measured by detecting the product of the enzymatic reaction. IC50 values are calculated from dose-response curves. Selectivity for PARP and tankyrase over other enzymes is assessed by screening against a panel of enzymes. These assays provide quantitative information on the potency and selectivity of JPI-547.

JPI-547 is tested on cultured cancer cells. Cells are treated with varying concentrations of JPI-547; PARP activity is assessed by measuring PAR polymer levels; tankyrase activity is assessed by measuring the levels of tankyrase substrates (e.g., AXIN); cell proliferation is assessed by MTT or CellTiter-Glo assays; apoptosis is assessed by Annexin V staining. These cell-based assays demonstrate the mechanism of action and antiproliferative activity of JPI-547.

JPI-547 has been evaluated in animal models of cancer. In these models, JPI-547 is administered orally at various doses; tumor growth inhibition is assessed by measuring tumor volume; PARP and tankyrase activity and downstream signaling are assessed in tumor tissue. These in vivo studies provide evidence for the efficacy of JPI-547.

Pharmacokinetic studies indicate that JPI-547 is orally active. The compound's absorption, distribution, metabolism, and excretion have been characterized in preclinical studies. JPI-547 achieves therapeutic concentrations in plasma and tissues following oral administration.

The toxicity profile of JPI-547 is consistent with that of other PARP inhibitors. Common adverse effects associated with PARP inhibitors include fatigue, nausea, anemia, and thrombocytopenia. The additional inhibition of tankyrase may result in a distinct toxicity profile that is being evaluated in preclinical and clinical studies.

[1]. Tricyclic derivative compound, method for preparing same, and pharmaceutical composition comprising same. Patent WO2016200101A2.

Nesupanib is a highly bioavailable, second-generation ribozyme inhibitor that inhibits poly(ADP-ribose) polymerase (PARP) types 1 (PARP1) and 2 (PARP2) as well as tankrase (TNK; TNKS; TANK) types 1 and 2, exhibiting potential chemosensitizing/radiosensitizing and antitumor activity. After oral administration, nesupanib selectively targets and binds to both PARP1/2 and TNK1/2 simultaneously. Inhibition of PARP activity prevents PARP-mediated single-strand DNA break repair (via the base excision repair pathway). This exacerbates the accumulation of DNA strand breaks, promotes genomic instability, and ultimately leads to apoptosis. This may enhance the cytotoxicity of DNA-damaging drugs. Inhibition of TNK activity blocks tankrase-mediated poly(ADP-ribose)ation of various target proteins, including multiple tumor suppressor factors. This may include blocking poly(ADP-ribose)ation and causing instability of the β-catenin negative regulator AXIN, thereby stabilizing AXIN. This blocks the Wnt/β-catenin signaling pathway and may inhibit the transcriptional activation of multiple Wnt/β-catenin target genes. This may suppress the proliferation of cancer cells with overactivated Wnt/β-catenin signaling. PARP catalyzes the post-translational ADP-ribosylation of nucleoproteins, which signal and recruit other proteins to repair damaged DNA. PARP can be activated by single-strand DNA breaks. PARP-mediated repair pathways are aberrant in multiple cancer cell types. TNK, a member of the PARP family, plays an important role in the regulation of the Wnt/β-catenin signaling pathway.
JPI-547 (Nesuparib, OCN-201) is a potent, orally active dual inhibitor of PARP1/2 and tankyrase1/2. The compound demonstrates antitumor activity in BRCA-deficient cancers. JPI-547 is being investigated for the treatment of cancer, neuropathic pain, neurodegenerative diseases, and cardiovascular diseases. The dual inhibition of PARP and tankyrase represents a novel approach to cancer therapy, potentially overcoming resistance to PARP inhibitor monotherapy. JPI-547 is not approved as a therapeutic agent and is used for research purposes only.

C23H24N6O

400.476263999939

400.201

2055357-64-5

2055357-64-5;2055357-65-6 (HCl);

124134768

White to off-white solid powder

1.8

2

6

3

30

733

0

O=C1C2CCCNC=2C2C=CC(=CC=2N1)CN1CCN(C2C=CC(C#N)=CN=2)CC1

GRPXLKXGJAGYSD-UHFFFAOYSA-N

InChI=1S/C23H24N6O/c24-13-17-4-6-21(26-14-17)29-10-8-28(9-11-29)15-16-3-5-18-20(12-16)27-23(30)19-2-1-7-25-22(18)19/h3-6,12,14,25H,1-2,7-11,15H2,(H,27,30)

6-[4-[(5-oxo-2,3,4,6-tetrahydro-1H-benzo[h][1,6]naphthyridin-8-yl)methyl]piperazin-1-yl]pyridine-3-carbonitrile

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

DMSO : ≥ 62.5 mg/mL (~156.06 mM)

Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.

---

Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)
*Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution.
Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)
Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil)
Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals).

View More

Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)]
*Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution.
Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline)
Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline)
Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline)
Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline
Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil)
Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)

---

Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium)
Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose
Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals).

View More

Oral Formulation 3: Dissolved in PEG400
Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose
Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose
Oral Formulation 6: Mixing with food powders

---

Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently.

---

 (Please use freshly prepared in vivo formulations for optimal results.)